EA042432B1 - SUBSTITUTED PHENYLSULFONYL PHENYLTRIAZOLTHIONES AND THEIR USE - Google Patents

Description

The field of technology to which the invention belongs

The present application relates to substituted phenylsulfonyl phenyltriazolethiones, pharmaceutical compositions containing these compounds, and methods for their use. These methods include, but are not limited to, prevention of aggregation or accumulation of neurotoxic proteins, improved clearance of these proteins, reduction of neuroinflammation, neuroprotective action, and treatment of pathological conditions associated with progressive accumulation of toxic protein compounds and/or neuroinflammation. These conditions include neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, Lewy body disease, Parkinson's disease with dementia, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple system atrophy, and progressive supranuclear palsy. In some embodiments, said condition is cancer, infection, Crohn's disease, heart disease, aging, traumatic brain injury (TBI), or a disease or condition associated with neuroinflammation.

Prior Art

It is estimated that neurodegenerative diseases such as Parkinson's disease or Alzheimer's disease affect more than 30 million people worldwide and are one of the leading causes of death in the elderly (Alzheimer Europe (2010), European Parkinson's Disease Association (2011)). The common hallmark of these neurological diseases is the chronic aggregation or accumulation of neurotoxic proteins and associated neuroinflammation. Compounds that prevent the overall progressive accumulation of these proteins and/or reduce neuroinflammation may provide a therapeutic advantage in the treatment of these disorders.

Brief description of the invention

In one aspect, the present invention describes a compound having formula (I)

where R ¹ , R ² and R ³ are each independently a hydrogen atom, a hydroxy group, a halogen, an optionally substituted C _1-4 alkyl, an optionally substituted C _1-4 alkoxy group, -CN, -C(O) R ^x , -C(O)OR ^x , -S(O)2R ^x or -NR ^y R ^z ;

R ^x , R ^y and R ^z are each independently H or optionally substituted C _1-4 alkyl; or R ^y and R ^z together with the nitrogen atom to which they are attached form an optionally substituted monocyclic heterocycloalkyl ring;

or a pharmaceutically acceptable salt thereof.

In some embodiments of formula (I), R ¹ is a hydrogen atom, an optionally substituted Ci-4-alkoxy group, or -NR ^y R ^z . In some embodiments of formula (I), R ¹ is a hydrogen atom. In some embodiments, R ¹ is a C1-4 alkoxy group that is unsubstituted or has one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl , hydroxyl, halogen, -NR ^f R ^g , cyano groups, nitro groups, C1-4 alkoxy groups, C _1-4 haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^g and -OC(O)NR ^f R ^g , where R ⁴ is H or C1-4 alkyl and R ^f and R ^g are each independently H, C1-4 alkyl, -C(O)C _1-4 alkyl, -C(O)OC _1-4 alkyl, or -S(O) ₂ C _1-4 alkyl. In some embodiments, R ¹ is -(OCH2CH2)P-O-CH2CH3 or -(OCH2CH2)p-O-CH3, where p is 0-10. In some embodiments, R ¹ is -OCH2CH ₂ -O-CH ₂ CH ₃ or -OCH ₂ CH ₂ OCH ₃ . In some embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z are each independently H or C 1-4 alkyl, where C 1-4 alkyl is unsubstituted or substituted with one or more substituents selected from the group, consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NR ^f R ^g , cyano group, nitro group, C1-4 alkoxy group, C _1-4 -haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^g and -OC(O)NR ^f R ^g , where R4 is H or C1-4 alkyl and R ^f and R ^g are each independently H, C1-4 alkyl, -C(O)C _1-4 alkyl, -C(O)OC _1-4 - alkyl or -8(O) ₂ C _1-4 -alkyl. In some embodiments, R ¹ is -NHCH2CH2OH or -N(CH2CH3)2. In some embodiments, R ¹ is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form an optionally substituted monocyclic heterocycloalkyl ring. In some embodiments, R ¹ is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form a monocyclic heterocycloalkyl ring selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl, where morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl are each unsubstituted or substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, _C2 -C6 alkynyl, hydroxyl, halogen, -NR ^f R ^g , cyano , nitro groups, C1-4-alkoxy groups, C1-4-haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O) NR ^f R ^g and -OC(O)NR ^f R ^g , where R ⁴ is H or C 1-4 alkyl and R ^f and R ^g are each independently H, C _1-4 alkyl, -C(O )C _1-4 - 1 042432 alkyl, -C(O)OC1_4-alkyl or -S(O) ₂ C _1-4 -alkyl. In some embodiments, R ¹ is morpholinyl, 4-methyl-piperazin-1-yl, piperidinyl, or pyrrolidinyl.

In some embodiments of formula (I), R ² is hydrogen, C1-4 alkyl, or substituted C1-4 alkyl. In some embodiments, R ² is C1-4 alkyl substituted with halo. In some embodiments, R ² is CF3. In some embodiments, R ² is methyl. In some embodiments, R ² is an optionally substituted C1-4 alkoxy group, -CN, or -NR ^y R ^z . In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z are each independently H or C1.4 alkyl, where C1.4 alkyl is unsubstituted or substituted with one or more substituents selected from the group, consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NRfR ^s , cyano group, nitro group, C1-4 alkoxy group, C ₁ . ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NRfR ^s and -OC(O)NRfR ^s where R ⁴ is is H or C1.4-alkyl; and Rf and ^{Rs are} each independently H, C1. ₄ -alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ . ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ² is —I(CH3)2. In some embodiments, R ² is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form an optionally substituted monocyclic heterocycloalkyl ring. In some embodiments, R ² is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form a monocyclic heterocycloalkyl ring selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl, where morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl are each unsubstituted or substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C ₂ -C6 alkenyl, C ₂ -C6 alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano- groups, nitro groups, C1.4-alkoxy groups, C1.4-haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O )NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C1. ₄ -alkyl and R ^f and R ^s are each independently H, C1.4-alkyl, -C(O)C1.4-alkyl, -C(O)OC1.4-alkyl, or -S(O)2C1. 4-alkyl. In some embodiments, R ² is morpholinyl. In some embodiments, R ² is -CN. In some embodiments, R ² is C1_4 alkoxy, unsubstituted or substituted with one or more substituents selected from the group consisting of C ₁ -C 6 alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, hydroxyl , halogen, -NR ^f R ^s , cyano groups, nitro groups, C1.4-alkoxy groups, C1.4-haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , - C(O)OR ⁴ , -C(O)NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C1. ₄ alkyl, and R ^f and R ^s are each independently H, C1.4 alkyl, -C(O)C1.4 alkyl, -C(O)OC1.4 alkyl, or -S(O)2C1. 4-alkyl. In some embodiments, R ² is -(OCH2CH ₂ ) _p -O-CH ₂ CH ₃ or -(OCH ₂ CH ₂ ) _p -OCH ₃ , where p is 0-10. In some embodiments, R ² is a methoxy group, -OCH ₂ CH ₂ -O-CH ₂ CH ₃ or -OCH ₂ CH ₂ OCH ₃ .

In some embodiments of formula (I), R ³ is halogen. In some embodiments, R ³ is chlorine. In some embodiments, R ³ is a hydrogen atom. In some embodiments, R ³ is C1.4 alkyl or substituted C1-4 alkyl. In some embodiments, R ³ is methyl. In some embodiments, R ³ is C1. ₄ -alkyl substituted with one or more halogen atoms. In some embodiments, R ³ is CF3. In some embodiments, R ³ is -CN. In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z are each independently H or C1. ₄ -alkyl, where C ₁ . _4- alkyl unsubstituted or substituted with one or more substituents selected from the group consisting of C ₁ -C 6 -alkyl, C ₂ -Cb-alkenyl, C ₂ -Cb-alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano- groups, nitro groups, C1.4-alkoxy groups, C1.4-haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C1. ₄ -alkyl and R ^f and R ^s are each independently H, C1.4-alkyl, -C(O)C1.4-alkyl, -C(O)OC1.4-alkyl, or -S(O)2C1. 4-alkyl. In some embodiments, R ³ is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form an optionally substituted monocyclic heterocycloalkyl ring. In some embodiments, R ³ is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form a monocyclic heterocycloalkyl ring selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl, where morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl are each unsubstituted or substituted with one or more substituents selected from the group consisting of C 1 -C 6 alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, C1.4-alkoxy groups, C1.4-haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C (O)NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C1. ₄ -alkyl and R ^f and R ^s are each independently H, C1.4-alkyl, -C(O)C1.4-alkyl, -C(O)OC1.4-alkyl or -S(O)2C1. 4-alkyl. In some embodiments, R ³ is morpholinyl. In some embodiments, R ³ is C1. ₄ alkoxy unsubstituted or substituted with one or more substituents selected from the group consisting of C ₁ -C 6 alkyl, C ₂ -C 6 alkenyl, C ₂ -C 6 alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, C ₁ . ₄ -alkoxy groups, C ₁ . ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C _1-4 alkyl and R ^f and R ^g are each independently H, C _1-4 -αkyl, -C(O)C _1-4 alkyl, -C( O)OC _1-4 -alkyl or -S(O) ₂ C _1-4 -alkyl. In some embodiments, R ³ is -(oCh ₂ CH ₂ )-O-CH ₂ CH ₃ or -(OCH ₂ CH ₂ ) _p -O-CH ₃ , where p is 0-10.

In some embodiments, the implementation of the compound having formula (I)is a compound from table. 1 or a pharmaceutically acceptable salt thereof.

In some embodiments, a compound having formula (I) is

or a pharmaceutically acceptable salt thereof.

In some embodiments, a compound having formula (I) is

O _s ,0

or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention describes a pharmaceutical composition comprising (a) at least one compound having formula (I) or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable excipient.

In some embodiments, the pharmaceutically acceptable excipient is a polymeric agent. In some embodiments, the pharmaceutically acceptable excipient is selected from the group consisting of carboxymethyl cellulose (CMC), hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC), hydroxypropyl methyl cellulose (HPMC), gelatin, gelatin hydrolyzate, sucrose, dextrose, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), vinyl pyrrolidone copolymers, pregelatinized starch, sorbitol and glucose; and polyacrylates. In some embodiments, the pharmaceutically acceptable excipient is selected from the group consisting of hydroxypropylmethyl cellulose (HPMC), polyvinylpyrrolidone (PVP), and kollidone. In some embodiments, the pharmaceutical composition is in the form of a dry spray dispersion (spray dry suspension, SDD).

The present application also describes a compound having formula I, or a pharmaceutically acceptable salt thereof, for use as a drug.

In some aspects, a compound having formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound having formula (I), is used to treat a condition caused by neurodegeneration or aggregation/accumulation of proteins such as alpha-synuclein, a -beta, tau, huntingtin or TAR DNA binding protein 43 (TDP43). In some embodiments, the pathological condition is a neurodegenerative disease or condition. In some embodiments, such a condition is Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Lewy body dementia, Parkinson's disease dementia, multiple system atrophy, Huntington's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, cancer, infection, Crohn's disease, heart, aging, or traumatic brain injury (TBI). In another aspect, a compound having formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound having formula (I), has a neuroprotective effect.

In some aspects, methods are described for treating a pathological condition caused by neurodegeneration or aggregation/accumulation of proteins, such as alpha-synuclein, a-beta, tau, huntingtin, or TDP43, comprising administering to a subject in need of such treatment, an effective amount of at least one compound, having formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound having formula (I). In some embodiments, said condition is a neurodegenerative disease or condition. In some embodiments, said condition is Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Lewy body dementia, Parkinson's disease dementia, multiple system atrophy, Huntington's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, cancer, inflammation, Crohn's disease, heart, aging, or traumatic brain injury (TBI).

In some aspects, the present invention describes the use of at least one compound having formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a pathological condition caused by neurodegeneration or protein accumulation. In some embodiments, said condition is Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Lewy body dementia, Parkinson's disease dementia, multiple system atrophy, Huntington's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, cancer, inflammation, Crohn's disease, heart, aging, or traumatic brain injury (TBI).

In another aspect, the present invention describes a method of preventing aggregation or accumulation or improving the clearance of a protease-resistant protein, comprising contacting the protease-resistant protein with an effective amount of at least one compound having formula (I), or a salt thereof, or described in the present text of the pharmaceutical composition, where the specified contact is carried out in vitro, ex vivo or in vivo. In some embodiments, the protease resistant protein is alpha-synuclein, a-beta, tau, huntingtin, or TDP43 proteins.

In another aspect, a compound having formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound having formula (I), is used to reduce neuroinflammation in a patient. In some embodiments, a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound of formula (I), is used to treat a disease or condition associated with neuroinflammation. In some embodiments, the present invention describes a method of reducing neuroinflammation in a patient, comprising administering to the subject an effective amount of at least one compound having formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein. In some embodiments, the present invention describes a method of treating a disease or condition associated with neuroinflammation, comprising administering to a subject an effective amount of at least one compound having formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein. In some embodiments, the present invention describes the use of at least one compound having formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for reducing neuroinflammation. In other embodiments, the present invention describes the use of at least one compound having formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a disease or condition associated with neuroinflammation.

Additional embodiments, features, and advantages of the compounds, compositions, methods, and uses described herein will be apparent from the detailed description below.

Brief description of the drawings

In FIG. 1A shows the 1H NMR spectrum of compound 1 in DMSO-dg (400 MHz).

In FIG. 1B shows the 2D NOESY spectrum of compound 1 in DMSO-dg (400 MHz) synthesized via route B.

In FIG. 1C shows a stretched 2D NOESY region of compound 1 in DMSO-dg (500 MHz) synthesized via pathway B.

In FIG. 1D shows the 2D NOESY spectrum of compound 1 in DMSO-d _g (400 MHz) synthesized via route C.

In FIG. 1E shows the HMWR spectrum of compound 1 in DMSO-d _g (400 MHz) synthesized via route C.

In FIG. 2A is a PXRD (powder x-ray diffraction) diagram of compound 1.

In FIG. 2B shows an overlay of PXRD charts for four different dry spray preparations of Compound 1.

In FIG. 3A shows an overlay of DSC (differential scanning calorimetry) and TGA (thermogravimetric analysis) thermograms for compound 1.

In FIG. ER and 3C show the TGA and DSC thermograms, respectively, for the Dry Spray Dispersion (SDD) #1.

In FIG. 3D and 3E show the TGA and DSC thermograms, respectively, for the Dry Spray Dispersion (SDD) #2.

In FIG. 3F and 3G show the TGA and DSC thermograms, respectively, for the Dry Spray Dispersion (SDD) #3.

In FIG. 3H and 3I show the TGA and DSC thermograms, respectively, for the Dry Spray Dispersion (SDD) #4.

In FIG. 4A shows the pharmacokinetic curves of Compound 1 in free base (CO) form and two dry spray dispersions of Compound 1 (SDD #1 and SDD #3).

In FIG. 4B shows area under the curve (AUC) versus dosage curves for Compound 1 in free base (CO) form and two dry spray dispersions of Compound 1 (SDD #1 and SDD #3).

- 4 042432

In FIG. 5A shows X-ray diffraction data for compound 1 (4-(4-((4-chloro-3(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione) .

In FIG. 5B shows X-ray diffraction data for an asymmetric cell of compound 1.

In FIG. 6 shows an X-ray powder diffraction (XRPD) plot of Compound 1.

In FIG. 7 shows the optical density of total alpha-synuclein deposits in (A) cortex, (B) hippocampus, and (C) striatum of L61 ASYN transgenic mice after intraperitoneal administration of compound 1 (1, 5, or 10 mg/kg per day) or vehicle (5 % DMSO + 20% Cremphor EL + 0.9% saline) for 1 month. Non-transgenic mice were used as controls and treated (ip) with Compound 1 (10 mg/kg per day) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month.

In FIG. 8 shows total alpha-synuclein deposits in representative cross-sectional images of the cortex, hippocampus, and striatum of L61 ASYN transgenic mice following intraperitoneal administration of Compound 1 (1, 5, or 10 mg/kg per day) or vehicle (5% DMSO+20% Cremphor EL +0.9% saline) for 1 month. Non-transgenic mice were used as controls and were injected (ip) with vehicle (5% DMSO+20% Cremphor EL+0.9% saline) for 1 month.

In FIG. 9 shows the optical density of deposits of insoluble alpha-synuclein (PK+resistant) in (A) cortex, (B) hippocampus and (C) striatum of L61 ASYN transgenic mice after intraperitoneal administration of Compound 1 (1.5 or 10 mg/kg per day). ) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month. Non-transgenic mice were used as controls and treated (ip) with Compound 1 (10 mg/kg per day) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month.

In FIG. 10 shows insoluble alpha-synuclein (PK+resistant) deposits on representative cross-sectional images of the neocortex, hippocampus, and striatum of L61 ASYN transgenic mice following intraperitoneal administration of compound 1 (1, 5, or 10 mg/kg per day) or vehicle (5% DMSO +20% Cremphor EL+0.9% saline) for 1 month. Non-transgenic mice were used as controls and were injected (ip) with vehicle (5% DMSO+20% Cremphor EL+0.9% saline) for 1 month.

In FIG. 11 shows a biochemical assessment of the level of monomeric ASYN in the brain in (A) frontal cortex and (B) hippocampus of L61 ASYN transgenic mice after intraperitoneal administration of compound 1 (1, 5 or 10 mg/kg per day) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month. Non-transgenic mice were used as controls and were injected (ip) with vehicle (5% DMSO+20% Cremphor EL+0.9% saline) for 1 month.

In FIG. 12 shows the absorbance of microtubule-associated protein 1A/1B light chain 3 (LC3) in (A) cortex, (B) hippocampus, and (C) striatum of L61 ASYN transgenic mice after intraperitoneal administration of Compound 1 (1.5 or 10 mg/kg per day) or carrier (5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month. Non-transgenic mice were used as controls and treated (ip) with Compound 1 (10 mg/kg per day) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month.

In FIG. 13 shows the level of LC3 immunostaining by IHC on representative images of transverse sections of the neocortex, hippocampus and striatum of L61 ASYN transgenic mice after intraperitoneal administration of compound 1 (1, 5 or 10 mg/kg per day) or vehicle (5% DMSO + 20% Cremphor EL +0.9% saline) for 1 month. Non-transgenic mice were used as controls and were injected (ip) with vehicle (5% DMSO+20% Cremphor EL+0.9% saline) for 1 month.

In FIG. 14 shows an assessment of grip strength in L61 ASYN transgenic mice after administration of compound 1 (5 or 10 mg/kg) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 3 months. Non-transgenic mice were used as controls and treated (ip) with Compound 1 (10 mg/kg per day) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 3 months.

In FIG. 15A shows the level of translocator protein (18 kDa) (TSPO) in representative cross-sectional images of the frontal cortex of L61 ASYN transgenic mice after administration of compound 1 (5 or 10 mg/kg) or vehicle (5% DMSO+20% Cremphor EL+0.9% saline). ) within 3 months. Non-transgenic mice were used as controls and treated (ip) with Compound 1 (10 mg/kg per day; results not shown) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 3 months.

In FIG. 15B shows the TSPO quantification of the images of FIG. 15A.

In FIG. 16 shows GFAP IHC staining in representative hippocampal images of L61 ASYN transgenic mice after intraperitoneal administration of compound 1 (1, 5 or 10 mg/kg per day) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month . Non-transgenic mice were used as controls and treated (ip) with Compound 1 (10 mg/kg per day; results not shown) or vehicle (5% DMSO+20% Cremphor EL+0.9% saline) for 3 months.

In FIG. 17 shows the absorbance by IHC staining of GFAP in the hippocampus of L61 ASYN transgenic mice after intraperitoneal administration of compound 1 (1.5 or 10 mg/kg per day) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 3 months. Non-transgenic mice were used as controls and treated (ip) with Compound 1 (10 mg/kg per day; results not shown) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month.

In FIG. 18 shows IHC staining of DAT in representative striatal cross-sectional images of L61 ASYN transgenic mice after administration of compound 1 (5 or 10 mg/kg) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 3 months. Non-transgenic mice were used as controls and treated (ip) with Compound 1 (10 mg/kg per day; results not shown) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline; results not shown) for 3 months.

In FIG. 19 shows striatal/reference densities ratio in IHC DAT staining on representative striatal and reference tract (cortex) images of L61 ASYN transgenic mice after administration of Compound 1 (5 or 10 mg/kg) or vehicle (5% DMSO+). 20% Cremphor EL + 0.9% saline) for 3 months. Non-transgenic mice were used as controls and treated (ip) with Compound 1 (10 mg/kg per day) or vehicle (5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month.

In FIG. 20 shows the quantification of TSPO immunofluorescent staining in representative brain sections of L41 APP transgenic mice after administration (ip once daily) of vehicle or Compound 1 at 5 mg/kg or vehicle for 70 days. Also shown are the results for non-transgenic mice treated with vehicle intraperitoneally once a day.

In FIG. 21 shows quantification of immunofluorescent staining for amyloid beta and 6E10 antibodies in representative brain sections of L41 APP transgenic mice after administration (ip once daily) of vehicle or Compound 1 at 5 mg/kg or vehicle for 70 days. Also shown are the results for non-transgenic mice treated with vehicle intraperitoneally once a day.

Detailed description of the invention

The present invention relates to substituted sulfonyl phenyl-2,4-dihydro-3H-1,2,4-triazole-3thiones, pharmaceutical compositions containing them, and methods of their use, including methods for the treatment of neurodegenerative diseases and other disorders in which there is an associated accumulation aggregates of toxic proteins.

Terms.

It should be understood that the compounds, compositions, methods, and uses described herein are not limited to the particular embodiments described. It should also be understood that the terminology used in this application is intended solely to describe particular embodiments and should not be construed as limiting in any way, since the scope of claims regarding the compounds, compositions, methods and uses described in this application is limited only by the attached Claims.

It should be noted that in the present text and in the appended claims, the use of the singular also includes the description of a plurality of subjects, unless the context clearly indicates otherwise. It should also be noted that any optional element may be excluded from the claims. This wording is used as a basis for the use of such exclusionary terms as exclusively, only, etc., in relation to the enumeration of the claimed elements or as negative signs.

As used in this text, the terms including and containing are used in an open, non-limiting sense.

The terms below have the meanings indicated unless otherwise noted. All terms for which no special definition is given have the meanings generally accepted in this field.

The term alkyl means a linear or branched alkyl (hydrocarbon) group containing from 1 to 12 carbon atoms in the chain. Examples of alkyl groups include methyl (Me), ethyl (Et), n-propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl (tBu), pentyl, isopentyl, t-pentyl, hexyl, isohexyl, and groups which in light standard knowledge in this field and the provisions of this application can be considered equivalent to any of the examples described below. In some cases, the alkyl groups are C ₁ . ₄ -alkyl.

Alkenyl means an unsaturated linear or branched hydrocarbon group containing the specified number of carbon atoms (eg 2-8 or 2-6 carbon atoms) and at least one olefinically unsaturated element (containing at least one carbon-carbon double bond). The alkenyl group may be in the cis or trans configuration (Z or E configuration) at the double bond(s). Alkenyl groups include (but are not limited to) ethenyl, propenyl (e.g. prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl (allyl), prop -2-en-2-yl) and butenyl

- 6 042432 (for example, but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-1-yl, but-2-en2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl).

Alkynyl means an unsaturated linear or branched hydrocarbon group containing the specified number of carbon atoms (eg 2-8 or 2-6 carbon atoms) and at least one acetylenically unsaturated element (containing at least one carbon-carbon triple bond). Alkynyl groups include, but are not limited to, ethynyl, propynyl (eg prop-1-yn-1-yl, prop-2-yn-1-yl) and butynyl (eg but-1-yn-1-yl , but-1-yn-3-yl, but-3yn-1-yl).

Aryl or Ar as used herein means an unsaturated aromatic carbocyclic group containing one ring (eg phenyl) or multiple fused rings (eg naphthyl or anthryl), the fused rings being carbocyclic and may or may not be aromatic, provided that at least one ring in the multi-fused ring structure is aromatic. Particular variants of aryl rings are rings containing from 6 to 14 carbon atoms in the cycle (C ₆ -C ₁₄ -aryl). An aryl group containing more than one ring, where at least one ring is non-aromatic, is connected to the rest of the molecule through a position located in the aromatic ring or in the non-aromatic ring. In one embodiment, an aryl group containing more than one ring, where at least one ring is non-aromatic, is connected to the rest of the molecule through a position located in the aromatic ring.

An alkoxy group means a -O-alkyl group, where alkyl is as defined above. The alkoxy group includes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, t-butoxy, n-pentoxy and other groups. The term alkoxy also means the groups alkenyl-O-, cycloalkyl-O-, cycloalkenyl-O- and alkynyl-O-, where alkenyl, cycloalkyl, cycloalkenyl and alkynyl are as defined above.

Cycloalkyl as used herein means and includes, unless otherwise indicated, saturated or partially unsaturated non-aromatic cyclic monovalent hydrocarbon structures containing the specified number of carbon atoms (for example, C ₃ -C 1o means from three to ten carbon atoms). Cycloalkyls can consist of a single ring, such as cyclohexyl, or multiple rings, such as adamantyl. Cycloalkyl containing more than one ring may be fused, spiro- or bridged, and combinations thereof are also possible. Particular examples of cycloalkyl groups are groups containing from 3 to 12 carbon atoms in the cycle. Preferred cycloalkyl is a cyclic hydrocarbon containing from 3 to 8 carbon atoms in the cycle (C ₃ -C ₈ -cycloalkyl), containing in the cycle from 3 to 6 carbon atoms (C ₃ -C ₆ -cycloalkyl) or containing in the cycle from 3 up to 4 carbon atoms (C ₃ -C ₄ -cycloalkyl). Examples of cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and the like.

Cyano or nitrile means a -CN group.

The term halo or halogen means fluorine, chlorine, bromine and iodine.

The term hydroxy or hydroxyl means a -OH group.

Heterocycloalkyl or heterocyclyl means a saturated or partially unsaturated group containing one ring or more fused rings, including conjugated, bridged or spiro-ring systems, and containing from 3 to 20 ring atoms, including from 1 to 10 heteroatoms. These ring atoms are selected from the group consisting of carbon, nitrogen, sulfur and oxygen, where in conjugated ring systems, one or more of the rings may be cycloalkyl, aryl, or heteroaryl, provided that the point of attachment is on a non-aromatic ring. In some embodiments, the nitrogen and/or sulfur atom(s) in the heterocyclic group are optionally oxidized to form N-oxide, -S(O)- or -S(O) ₂ moieties. Examples of heterocycloalkyls include (but are not limited to) azetidine, oxetane, tetrahydrofuran, pyrrolidine, piperazine, piperidine, morpholine, thiomorpholine, 1,1-dioxothiomorpholinyl, dihydroindole, indazole, quinolysine, imidazolidine, imidazoline, indoline, 1,2,3, 4-tetrahydroisoquinoline, thiazolidine, and the like. In some cases, heterocycloalkyl groups are 4-, 5-, or 6-membered rings. In some cases, heterocycloalkyl contains a conjugated phenyl ring.

Heteroaryl as used herein means an unsaturated aromatic cyclic group containing from 1 to 14 carbon atoms in the ring and at least one heteroatom in the ring, including (but not limited to) heteroatoms such as nitrogen, oxygen and sulfur. The heteroaryl group may contain one ring (eg, pyridyl, furyl) or multiple fused rings (eg, indolizinyl, benzothienyl), while the fused rings may be carbocyclic or may contain one or more heteroatoms in the ring, and they may also be aromatic or non-aromatic provided that at least one ring in the multi-fused ring structure is aromatic and contains at least one heteroatom in the ring, and provided that the point of attachment to the rest of the molecule is in an aromatic ring containing at least one heteroatom. The heteroaryl group may be attached to the rest of the molecule through a carbon atom in the ring or through a heteroatom in the ring. Particular examples of heteroaryl groups are 5-14-membered rings containing in the cycle from 1 to 12 carbon atoms and from 1 to 6 heteroatoms independently selected from nitrogen, oxygen and sulfur atoms, 5-10-membered rings containing in the cycle from 1 up to 8 carbon atoms and from 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur atoms, or 5, 6 or 7-membered rings containing from 1 to 5 carbon atoms and from 1 to 4 heteroatoms independently selected from atoms of nitrogen, oxygen and sulfur. In one embodiment, particular examples of heteroaryl groups are monocyclic aromatic 5-, 6-, or 7-membered rings containing 1 to 6 carbon atoms and 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur atoms. In another embodiment, specific examples of heteroaryl groups are polycyclic aromatic rings containing from 1 to 12 carbon atoms and from 1 to 6 heteroatoms independently selected from nitrogen, oxygen and sulfur atoms.

Oxo means a (=O) or (O) group.

As used herein, the term substituted, when used as a modifier of a particular group or radical, may also mean that one or more hydrogen atoms in the specified group or radical are each independently replaced by the same or different substituent groups, as defined below.

In addition to the groups described in this text for individual terms, substituent groups to replace one or more hydrogen atoms (any two hydrogen atoms on one carbon atom can be replaced by =O, =NR ⁷⁰ , =N-OR ⁷⁰ , = N2 or =S) at saturated carbon atoms in the indicated group or radical are, unless otherwise indicated, -R ⁶⁰ , halogen, =O, -OR ⁷⁰ , -SR ⁷⁰ , -NR ⁸⁰ R ⁸⁰ , trihalomethyl, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)R ⁷⁰ , -S(O)2R ⁷⁰ , -S(O)2O'M ⁺ , -S(O)2OR ⁷⁰ , -OS(O)2R ⁷⁰ , -OS(O)2O'M ⁺ , -OS(O)2OR ⁷⁰ , -P(O)(O')2(M ⁺ )2, -P(O)( OR ⁷⁰ )O'M ⁺ , -P(O)(OR ⁷⁰ )2, -C(O)R ⁷⁰ , -C(S)R ⁷⁰ , -C(NR ⁷⁰ )R ⁷⁰ , -C(O)O 'M ⁺ , -C(O)OR ⁷⁰ , -C(S)OR ⁷⁰ , -C(O)NR ⁸⁰ R ⁸⁰ , -C(NR ⁷⁰ )NR ⁸⁰ R ⁸⁰ , -OC(O)R ⁷⁰ , - OC(S)R ⁷⁰ , -OC(O)O'M ⁺ , -OC(O)OR ⁷⁰ , -OC(S)OR ⁷⁰ , -NR ⁷⁰ C(O)R ⁷⁰ , -NR ⁷⁰ C(S) R ⁷⁰ , -NR ⁷⁰ C(O)O ^- M ⁺ , -NR ⁷⁰ C(O)OR ⁷⁰ , -NR ⁷⁰ C(S)OR ⁷⁰ , -NR ⁷⁰ C(O)NR ⁸⁰ R ⁸⁰ , -NR ⁷⁰ C(NR ⁷⁰ )R ⁷⁰ and NR ⁷⁰ C(NR ⁷⁰ )NR ⁸⁰ R ⁸⁰ , where R ⁶⁰ is selected wounds from the group consisting of optionally substituted alkyl, cycloalkyl, heterocycloalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl, each R ⁷⁰ independently represents a hydrogen atom or R ⁶⁰ ; each R ⁸⁰ is independently R ⁷⁰ or alternatively two R ^80s together with the nitrogen atom to which they are attached form a 3-, 4-, 5-, 6-, or 7-membered heterocycloalkyl which may optionally include from 1 to 4 identical or various additional heteroatoms selected from the group consisting of O, N and S, of which N may have substituents -H, C1-C4-alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ _ 4-alkyl or -S(O) ₂ C ₁ . ₄ -alkyl; and each M ⁺ is a counterion with a unit positive charge. Each M ⁺ may independently represent, for example, an alkali metal ion such as K ⁺ , Na ⁺ , Li ⁺ ; an ammonium ion such as +N(R ⁶⁰ )4; or an alkaline earth metal ion such as [Ca ²⁺ ]0. ₅ , [Mg ²⁺ ]0.5 or [Ba ²⁺ ]0. ₅ (subscript 0.5 means that one of the counterions for such divalent alkaline earth ions may be the ionized form of the compound described in this text, and another typical counterion, such as chloride, or the two ionized compounds described in this text, may serve as counterions for such divalent alkaline earth metal ions, or a doubly ionized compound described herein may serve as a counterion for such divalent alkaline earth metal ions). As particular examples, -NR ⁸⁰ R ⁸⁰ includes -NH2, -NH-alkyl, N-pyrrolidinyl, N-piperazinyl, 4-N-methylpiperazin-1-yl and N-morpholinyl.

In addition to the substituent groups described herein in terms of individual terms, substituent groups for replacing hydrogen atoms on nitrogen atoms in substituted heterocycloalkyl groups are, unless otherwise indicated, -R ⁶⁰ , -O ^- M ⁺ , -OR ⁷⁰ , -SR ⁷⁰ , -S ^- M ⁺ , -NR ⁸⁰ R ⁸⁰ , trihalomethyl, -CF3, -CN, -NO, -NO2, -S(O)R ⁷⁰ , -S(O)2R ⁷⁰ , -S( O)2O'M ⁺ , -S(O)2OR ⁷⁰ , -OS(O)2R ⁷⁰ , -OS(O)2O'M ⁺ , -OS(O)2OR ⁷⁰ , -P(O)(O') 2(M ⁺ )2, -P(O)(OR ⁷⁰ )O'M ⁺ , -P(O)(OR ⁷⁰ )(OR ⁷⁰ ), -C(O)R ⁷⁰ , -C(S)R ⁷⁰ , -C(NR ⁷⁰ )R ⁷⁰ , -C(O)OR ⁷⁰ , -C(S)OR ⁷⁰ , -C(O)NR ⁸⁰ R ⁸⁰ , -C(NR ⁷⁰ )NR ⁸⁰ R ⁸⁰ , -OC( O)R ⁷⁰ , -OC(S)R ⁷⁰ , -OC(O)OR ⁷⁰ , -OC(S)OR ⁷⁰ , -NR ⁷⁰ C(O)R ⁷⁰ , -NR ⁷⁰ C(S)R ⁷⁰ , - NR ⁷⁰ C(O)OR ⁷⁰ , -NR ⁷⁰ C(S)OR ⁷⁰ , NR ⁷⁰ C(O)NR ⁸⁰ R ⁸⁰ , -NR ⁷⁰ C(NR ⁷⁰ )R ⁷⁰ and -NR ⁷⁰ C(NR ⁷⁰ )NR ⁸⁰ R ⁸⁰ where R ⁶⁰ , R ⁷⁰ , R ⁸⁰ and M ⁺ correspond to the above definitions. When a heterocycloalkyl group is substituted, unless otherwise limited by the definition of a substituent on a heterocycloalkyl, such groups may be substituted with 1-5 or 1-3 substituents selected from alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl , carboxyl ester, thioaryloxy group, thioheteroaryloxy group, thioheterocyclooxy group, thiol, thioalkoxy group, substituted thioalkoxy group, aryl, aryloxy group, heteroaryl, heteroaryloxy group, heterocyclyl, heterocyclooxy group, hydroxyamino group, alkoxyamino groups, nitro groups, sulfonylamino groups, -S(O)-alkyl, -S(O)- substituted alkyl, -S(O)- 8 042432 aryl, -S(O)-heteroaryl, -S(O) -heterocyclyl, -S(O) ₂ -alkyl, -S(O) ₂ -substituted alkyl, -S(O) ₂ -aryl,

-S(O) ₂ -heteroaryl; and -S(O) ₂ -heterocyclyl.

It should be understood that when a group is indicated as substituted, it may be substituted with one or more substituents and the substituents may be in any or all positions of the system, as long as the valency rules allow. In some embodiments, the group that is substituted has 1, 2, 3, or 4 substituents, 1, 2, or 3 substituents, 1 or 2 substituents, or 1 substituent.

The term optionally substituted, unless otherwise indicated, means that a group may be unsubstituted or substituted with one or more (e.g., 1, 2, 3, 4, or 5) substituents listed for that group, where the substituents may be the same or different. In one embodiment, the optionally substituted group has one substituent. In another embodiment, the optionally substituted group has two substituents. In another embodiment, the optionally substituted group has three substituents. In another embodiment, the optionally substituted group has four substituents. In some embodiments, an optionally substituted group has 1-2, 2-5, 3-5, 2-3, 2-4, 3-4, 1-3, 1-4, or 1-5 substituents. In one embodiment, the optionally substituted group has no substituents.

Unless otherwise indicated, the nomenclature of substituents not explicitly listed in this text is formed by indicating the terminal fragment of the functional group, followed by the name of the functional group at the point of attachment.

For all groups described in this text containing one or more substituents, it is obviously understood that such groups do not contain any substituents or sets of substituents that are not sterically and/or unattainable synthetically. In addition, the compounds of the present invention include all stereochemical isomers resulting from the introduction of substituents in these compounds.

The term pharmaceutically acceptable salt means a salt that is acceptable for administration to a patient, such as a mammal (salts with counterions having mammalian acceptable safety in the selected dosage regimen). Such salts may be derived from pharmaceutically acceptable inorganic or organic bases and pharmaceutically acceptable inorganic or organic acids. The term pharmaceutically acceptable salt means pharmaceutically acceptable salts of a compound that are derived from a wide range of organic and inorganic counterions well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like ions. .; and, in case the molecule contains a basic functional group, organic or inorganic acid salts such as hydrochloride, hydrobromide, formate, tartrate, besylate, mesylate, acetate, maleate, oxalate, and the like.

The term solvate means a complex formed by combining solvent molecules with solute molecules or ions. The solvent may be an organic compound, an inorganic compound, or a mixture thereof. Some examples of solvents include, but are not limited to, methanol, K,K-dimethylformamide, tetrahydrofuran, dimethyl sulfoxide, and water. When the solvent is water, the resulting solvate is a hydrate.

The term stereoisomer and stereoisomers refer to compounds that have the same atomic bonding order but a different arrangement of atoms in space. Stereoisomers include cis-trans isomers, E and Z isomers, enantiomers and diastereomers. Compounds having asymmetric centers may exist as one or more enantiomeric forms, one or more diastereomeric forms, one or more atropisomeric forms, and as mixtures thereof in any ratio.

Any formula given herein also includes any of the hydrate, solvate, amorphous, and polymorphic forms of such compounds, and mixtures thereof, even if such forms are not explicitly listed.

Any formula given in this text also includes unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have structures that correspond to the formulas depicted in this text, except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that may be included in the compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine and iodine, such as ² H, ³ H, 11 C, ¹³ C, ¹⁴ C, ¹⁵ N , ¹⁸ O, ¹⁷ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ³⁶ Cl, and ¹²⁵ I, respectively. Such isotopically labeled compounds can be used in studies of metabolism (e.g. ¹⁴ C), reaction kinetics (e.g. ² H or ³ H), detection or imaging techniques (such as positron emission tomography (PET) or single photon emission computed tomography (SPECT). )), including analysis of the distribution of the drug or substrate in tissues, or in radiation therapy of patients. In some embodiments, compounds labeled with isotopes ¹⁸ F or 11C are used in PET or SPECT studies. PET and SPECT examinations can be performed as described, for example, in Brooks, DJ, Positron Emission Tomography and

- 9 042432

Single-Photon Emission Computed Tomography in Central Nervous System Drug Development, NeuroRx 2005, 2(2), 226-236 and references therein. In addition, substitution for heavier isotopes such as deuterium (ie ² H) may provide certain therapeutic advantages due to higher metabolic stability, such as increased half-life or reduced dosage required. In general, isotopically labeled compounds and their prodrugs can be prepared by following the synthetic procedures and methods described in the Schemes or Examples in this text, while replacing an isotopically labeled reagent with an isotopically labeled reagent.

For a more precise description, some of the quantitative values given in this text do not contain the term approximately. It should be understood that, whether the term is approximately explicitly present or not, each quantitative value given in this text refers to the explicitly stated value, as well as to the approximation to such explicitly indicated value that is reasonable to assume based on standard knowledge. in the art, including equivalents and approximations for such quantity due to experimental and/or measurement errors. When a yield is given as a percentage, that yield means the mass of the substance for which the yield is given, relative to the maximum amount of that substance that could be obtained under the stated stoichiometric conditions.

Percentage concentrations are by weight unless otherwise indicated.

Unless otherwise indicated, all technical and scientific terms used in this text have the meanings known to those skilled in the art. Methods and materials are described in this text; however, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the compounds or compositions described herein. All publications cited in this text are incorporated by reference in the text for the purpose of disclosing and describing the methods and/or materials in connection with which these publications are cited.

Unless otherwise indicated, the methods and techniques described in this text are implemented according to well-known methods in this field, as described in various general and more specific sources cited in this text. See, for example, Loudon, Organic Chemistry, 4th edition, New York: Oxford University Press, 2002, p. 360-361, 1084-1085; Smith and March, March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition, Wiley-Interscience, 2001.

The nomenclature used in this text to create the names of the compounds in question is illustrated in the examples in this text. This nomenclature uses the commercially available ChemBioDraw Ultra 13.0.2.3021 program (CambridgeSoft, Cambridge, Mass.).

It should be understood that particular features of the compounds, compositions, methods, and uses described herein, which, for clarity, are described in the context of separate embodiments, may also be combined in combination in one embodiment. Conversely, the various features of the compounds, compositions, methods, and uses described herein for brevity in the context of one embodiment may also be used alone or in any suitable subcombination. All combinations of embodiments relating to chemical groups represented by variables are covered by this application and are described in this text as if each and every combination were separately and explicitly described, provided that such combinations cover compounds that are stable (t i.e. compounds that can be isolated, characterized and tested for biological activity). In addition, all subcombinations of chemical groups listed in the embodiments describing each variable are also covered by this application and are described in the same way as if each and every such subcombination of chemical groups were separately and explicitly described in this text.

Connections.

Compounds and their salts (such as pharmaceutically acceptable salts) are described in detail in the present text, including the abstract and claims. Also described are the uses of all compounds described herein, including salts and solvates of the compounds described herein, as well as methods for preparing such compounds. Any compound described herein may also be referred to as a drug.

In one aspect, compounds having formula (I) are described.

where R ¹ , R ² and R ³ each independently represent a hydrogen atom, a hydroxy group, a halogen, an optionally substituted C1-4 alkyl, an optionally substituted C1-4 alkoxy group, -CN, -C(O)R ^x , -C(O)OR ^x , -S(O)2R ^x or -NR ^y R ^z ;

R ^x , R ^y and R ^z are each independently H or optionally substituted C _1-4 alkyl

- 10 042432 or R ^y and R ^z together with the nitrogen atom to which they are attached form an optionally substituted monocyclic heterocycloalkyl ring;

or a pharmaceutically acceptable salt thereof.

In some embodiments, when a group is described as optionally substituted, said group is unsubstituted or substituted with one or more substituents selected from the group consisting of oxo, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ -alkynyl, halogen, -CN, -OR ⁴ , -SR ⁴ , -NR5R ⁶ , -NO2, -C=NH(OR ⁴ ), -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ⁵ R ⁶ , -OC(O)NR ⁵ R ⁶ , -NR ⁴ C(O)R ⁵ , -NR ⁴ C(O)OR ⁵ , -NR ⁴ C(O)NR ⁵ R ⁶ , -S(O)R ⁴ , -S(O)2R ⁴ , -NR ⁴ S(O)R ⁵ , -C(O)NR ⁴ S(O)R ⁵ , -NR ⁴ S(O)2R ⁵ , -C(O)NR ⁴ S(O)2R ⁵ , -S(O)NR ⁵ R ⁶ , -S(O)2NR ⁵ R ⁶ , -P(O )(OR ⁵ ) (OR ⁶ ), C3-C6-cycloalkyl, 3-12-membered heterocyclyl, 5-10-membered heteroaryl, C6-C14-aryl, -(C1-C3-alkylene) CN, -(C1- C3-alkylene) OR ⁴ , -(C1-C3-alkylene) SR ⁴ , -(C1- _C3- alkylene) NR5R ⁶ , -(C1-C3-alkylene) CF3, -(C1-C3-alkylene) NO2, -C =NH(OR ⁴ ), -(Ci-C3 alkylene) C(O)R ⁴ , -(C1-C3-alkylene)OC(O)R ⁴ , -(C1-C3-alkylene) C(O)OR ⁴ , -(C1-C3-alkylene) C(O)NR ⁵ R ⁶ , -(C1-C3-alkylene) OC(O)NR ⁵ R ⁶ , -(C1-C3-alkylene) NR ⁴ C(O)R ⁵ , -(C1-C3-alkylene) NR ⁴ C(O)OR ⁵ , -(C1-C3 alkylene) NR ⁴ C(O)NR ⁵ R ⁶ , -(C1-C3-alkylene)S(O)R ⁴ , -(C1-C3-alkylene)S(O)2R ⁴ , -(C1-C3-alkylene)NR ⁴ S(O)R ⁵ , - C(O)(C1-C3-alkylene)NR ⁴ S(O)R ⁵ , -(C1-C3-alkylene)NR ⁴ S(O)2R ⁵ , -(C1-C3-alkylene)C(O)NR ⁴ S(O)2R ⁵ , -(C1-C3 alkylene)S(O)NR ⁵ R ⁶ , -(C1-C3-alkylene)S(O)2NR ⁵ R ⁶ , -(C1-C ₃ -alkylene)P (O)(OR ⁵ )(OR ⁶ ), -(C1-C3-alkylene)(C3C6-cycloalkyl), -(C1-C3-alkylene)(3-12 membered heterocyclyl), -(C1-C3-alkylene )(5-10 membered heteroaryl) and -(C1-C3-alkylene)(C6-C14-aryl) wherein said one or more substituents are each independently unsubstituted or have one or more additional substituents selected from the group consisting of from halogen, oxo group, -OR ⁷ , -NR ⁷ R ⁸ , -C(O)R ⁷ , -CN, -S(O)R ⁷ , -S(O)2R ⁷ , -P(O)( OR ⁷ )(OR ⁸ ), -(C1-C3-alkylene)OR ⁷ , -(C1-C ₃ -alkylene)NR ⁷ R ⁸ , -(C1-C3-alkylene)C(O)R ⁷ , -( C1-C ₃ alkylene)S(O)R ⁷ , -(C1-C3 alkylene)S(O)2R ⁷ , -(C1-C ₃ alkylene)P(O)(OR ⁷ )(OR ⁸ ), C ₃ -С ₈ - cycloalkyl, C ₁ -C ₆ alkyl and C ₁ -C ₆ alkyl substituted with oxo, —OH, or halogen;

where each R ⁴ is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C14 aryl, 5-6 membered heteroaryl, or 3-6 -membered heterocyclyl, where C1-C6-alkyl, C2-C6-alkenyl, C2-C6-alkynyl, C3-C6-cycloalkyl, C6-C14-aryl, 5-6-membered heteroaryl and 3-6-membered heterocyclyl are independently unsubstituted or substituted with halogen, oxo, -CN, -OR ⁹ , -NR ⁹ R ¹⁰ , -P(O)(OR ⁹ )(OR ¹⁰ ), phenyl, phenyl substituted with halogen, C1-C ₆ alkyl, or C ₁ -C ₆ -alkyl substituted with halogen, -OH or oxo;

R ⁵ and R ⁶ are each independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C14 aryl, 5-6 membered heteroaryl, or 3 -6-membered heterocyclyl, where C1-C6-alkyl, C2-C6-alkenyl, C2-C6-alkynyl, C3-C6-cycloalkyl, C6-C14-aryl, 5-6-membered heteroaryl and 3-6-membered heterocyclyl each independently unsubstituted or substituted with halogen, oxo, -CN, -OR ⁹ , -NR ⁹ R ¹⁰ , C1-C ₆ alkyl, or C ₁ -C ₆ alkyl substituted with halogen, -OH, or oxo; And

R ⁷ , R ⁸ , R ⁹ and R ¹⁰ are each independently a hydrogen atom, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkyl, substituted with one or more halogens, C ₂ -C ₆ alkenyl substituted with one or more halogens, or C ₂ -C ₆ alkynyl substituted with one or more halogens.

In some embodiments of formula (I), R ¹ , R ² and R ³ are each independently hydrogen, hydroxy, halogen, optionally substituted C1.4 alkyl, optionally substituted C1.4 alkoxy, or —NR ^y R ^z In some cases, for each of R ¹ , R ² and R ³ C1. ₄ -alkyl or C ₁ . _{The 4-} alkoxy groups are substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NR ^f R ^g , cyano, nitro, C1.4-alkoxy, and C1.4-haloalkoxy, where R ^f and R ^g are each independently H, C1. ₄ -alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ - ₄ alkyl or -S(O) ₂ C ₁ . ₄ -alkyl.

In some embodiments, one or more of R ¹ , R ^{2 ,} or R ³ is C1.4-alkyl which is unsubstituted or has one or more substituents selected from the group consisting of halo, -CN, -OR ⁴ , -SR ⁴ , -NR ⁵ R ⁶ , -NO2, -C=NH(OR ⁴ ), -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O )NR ⁵ R ⁶ , -OC(O)NR ⁵ R ⁶ , -NR ⁴ C(O)R ⁵ , -NR ⁴ C(O)OR ⁵ , -NR ⁴ C(O)NR ⁵ R ⁶ , -S (O)R ⁴ , -S(O)2R ⁴ , -NR ⁴ S(O)R ⁵ , -C(O)NR ⁴ S(O)R ⁵ , -NR ⁴ S(O)2R ⁵ , -C (O)NR ⁴ S(O)2R ⁵ , -S(O)NR ⁵ R ⁶ , -S(O)2NR ⁵ R ⁶ , -P(O)(OR ⁵ )(OR ⁶ ), C3-C ₆ -cycloalkyl, 3-12-membered heterocyclyl, 5-10-membered heteroaryl and C ₆ -C ₁₄ -aryl;

where R ⁴ is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C14 aryl, 5-6 membered heteroaryl, or 3-6- membered heterocyclyl, wherein C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C14 aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl are independently optionally substituted halogen, oxo, -CN, -OR ⁹ , -NR ⁹ R ¹⁰ , -P(O)(OR ⁹ )(OR ¹⁰ ), phenyl optionally substituted with halogen, or C1-C ₆ -alkyl optionally substituted with halogen , -OH or oxo group;

R ⁵ and R ⁶ are each independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C14 aryl, 5-6 membered heteroaryl, or 3 -6-membered heterocyclyl, where C1-C6-alkyl, C2-C6-alkenyl, C2-C6-alkynyl, C3-C6-cycloalkyl, C6-C14-aryl, 5-6-membered heteroaryl and 3-6-membered heterocyclyl independently optionally substituted with halogen, oxo, -CN, -OR ⁹ , -NR9R ¹⁰ or C1-C ₆ alkyl optionally substituted with halogen, -OH or oxo; And

R ⁹ and R ¹⁰ each independently represent a hydrogen atom, C ₁ -C ₆ -alkyl, C ₂ -C ₆ -alkenyl,

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C ₂ -C6 alkynyl, C1 -C6 alkyl substituted with one or more halogens, C ₂ -C ₆ alkenyl substituted with one or more halogens, or C ₂ -C ₆ alkynyl substituted with one or more halogens.

In some embodiments, one or more of R ¹ , R ² or R ³ is a C1.4 alkoxy group that is unsubstituted or has one or more substituents selected from the group consisting of C1-C6 alkyl, C2 -C6-alkenyl, C2-C6-alkynyl, halogen, -CN, -OR ⁴ , -SR ⁴ , -NR5R ⁶ , -NO2, -C=NH(OR ⁴ ), -C(O)R ⁴ , -OC (O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ⁵ R ⁶ , -OC(O)NR ⁵ R ⁶ , -NR ⁴ C(O)R ⁵ , -NR ⁴ C(O )OR ⁵ , -NR ⁴ C(O)NR ⁵ R ⁶ , -S(O)R ⁴ , -S(O)2R ⁴ , -NR ⁴ S(O)R ⁵ , -C(O)NR ⁴ S (O)R ⁵ , -NR ⁴ S(O)2R ⁵ , -C(O)NR ⁴ S(O)2R ⁵ , -S(O)NR ⁵ R ⁶ , -S(O)2NR ⁵ R ⁶ , -P(O)(OR ⁵ )(OR ⁶ ), C3-C ₆ cycloalkyl, 3-12 membered heterocyclyl, 5-10 membered heteroaryl and C ₆ -C ₁₄ aryl;

where R ⁴ independently represents a hydrogen atom, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C14 aryl, 5-6 membered heteroaryl, or 3-6- membered heterocyclyl, wherein C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C14 aryl, 5-6 membered heteroaryl and 3-6 membered heterocyclyl are independently optionally substituted halogen, oxo, -CN, -OR ⁹ , -NR9R ¹⁰ , -P(O)(OR ⁹ )(OR ¹⁰ ), phenyl optionally substituted with halogen, or C1-C ₆ -alkyl optionally substituted with halogen, - OH or oxo group; And

R ⁵ and R ⁶ are each independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C14 aryl, 5-6 membered heteroaryl, or 3 -6-membered heterocyclyl, where C1-C6-alkyl, C2-C6-alkenyl, C2-C6-alkynyl, C3-C6-cycloalkyl, C6-C14-aryl, 5-6-membered heteroaryl and 3-6-membered heterocyclyl independently optionally substituted with halogen, oxo, -CN, -OR ⁹ , -NR ⁹ R ¹⁰ or C1-C ₆ -alkyl optionally substituted with halogen, -OH or oxo; And

R ⁹ and R ¹⁰ are each independently a hydrogen atom, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkyl substituted with one or more halogens, C ₂ -C ₆ alkenyl substituted with one or more halogens, or C 2 -C 6 alkynyl substituted with one or more halogens.

In some embodiments, R ¹ is hydrogen, hydroxy, halogen, optionally substituted C1.4 alkyl, optionally substituted C1.4 alkoxy, or —NR ^y R ^z . In some embodiments, R ¹ is a hydrogen atom. In some embodiments, R ¹ is hydroxyl. In some embodiments, R ¹ is halogen. In some embodiments, R ¹ is chlorine. In some embodiments, R ¹ is fluorine. In other embodiments, R ¹ is bromine or iodine. In some embodiments, R ¹ is an optionally substituted C1. ₄ -alkyl. In some embodiments, R ¹ is C1.4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halo, NRfR ^s , cyano, nitro, C1. ₄ alkoxy groups and C ₁ . ₄ -haloalkoxy groups, where Rf and R ^s are each independently H, C1.4-alkyl, -C(O)C1.4-alkyl, -C(O)OC1.4-alkyl, or -S(O) 2C1.4alkyl. In some embodiments, R ¹ is C 1-4 alkyl substituted with one or more halogen atoms. In some embodiments, R ¹ is -CF3, -(CH2)F, -CHF2, CH2Br, -CH2CF3, -CH2CHF2, or -CH2CH2F. In some embodiments, R ¹ is CF3. In some embodiments, R ¹ is unsubstituted C ₁ . ₄ -alkyl. For example, in some embodiments, R ¹ is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, or tert-butyl.

In other embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z , together with the nitrogen atom to which they are attached, form an optionally substituted monocyclic heterocycloalkyl ring. In some embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z together with the nitrogen atom to which they are attached form an optionally substituted 5-12 membered heterocycloalkyl ring. In some embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z , together with the nitrogen atom to which they are attached, form an optionally substituted 5-6 membered heterocycloalkyl ring. In some embodiments, R ¹ is morpholinyl. In some embodiments, R ¹ is morpholinyl substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, C1. ₄ -alkoxy groups, C ₁ . ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C1.4 alkyl and R ^f and R ^s are each independently H, C1. ₄ -alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ . ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ¹ is piperazinyl. In some embodiments, R ¹ is piperazinyl substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, C1. ₄ -alkoxy groups, C ₁ . ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C1.4 alkyl and R ^f and R ^s are each independently H, C1. ₄ -alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ . ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ¹ is piperidinyl. In some embodiments, R ¹ is piperidinyl substituted with one or more substituents selected from the group consisting of C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, C _1-4 alkoxy groups, C ₁ - ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NRfR ^g and -OC(O)NRfR ^s where R ⁴ is H or C1-4 alkyl and Rf and R ^g are each independently H, C1-4 -alkyl, -C(O)C ₁ - ₄ -alkyl, -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ _ ₄ -alkyl. In some embodiments, R ¹ is pyrrolidinyl. In some embodiments, R ¹ is pyrrolidinyl substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NRfR ^g , cyano, nitro groups, C1-4 alkoxy groups, C ₁ - ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O) NR ^f R ^g and -OC(O)NR ^f R ^g , where R ⁴ is H or C1-4 alkyl and Rf and R ^g are each independently H, C1-4 alkyl, -C(O)C ₁ - ₄ -alkyl, -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl.

In some embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z are each independently H or optionally substituted C1-4 alkyl. In some embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z are each H. In some embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z are each optionally substituted C1-4-alkyl. In some embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z are each optionally substituted C1-4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NR ^f R ^g , cyano groups, nitro groups, C1-4 alkoxy groups, C ₁ - ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NRfR ^g and -OC(O)NRfR ^g , where R ⁴ is H or C1-4 alkyl and Rf and R ^g are each independently H, C1-4 alkyl, -C( O)C ₁ - ₄ -alkyl, -C (O) OC ₁ - ₄ -alkyl or -S (O) ₂ C ₁ _ ₄ -alkyl. In some embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z are each optionally substituted C1-4 alkyl. In some embodiments, R ¹ is -N(CH2)2 or -N(CH2CH ₃ ) ₂ . In some embodiments, R ¹ is -NR ^y R ^z , where R ^y and R ^z are each unsubstituted C1-4 alkyl or C 1-4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NR ^f R ^g , cyano groups, nitro groups, _C1-4 alkoxy groups and _C1-4 haloalkoxy groups, wherein Rf and R ^g are each independently H, C1-4 alkyl, -C( O)C1-4-alkyl, -C(O)OC1-4-alkyl or -S(O)2C1_4-alkyl. In some embodiments, R ¹ is -NR ^y R ^z , where one of R ^y and R ^z is H and the other is unsubstituted C1-4 alkyl. In other embodiments, R ¹ is -NR ^y R ^z , where one of R ^y and R ^z is H and the other is C 1-4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NR ^f R ^g , cyano groups, nitro groups, C 1-4 alkoxy groups and C ₁ - ₄ haloalkoxy groups, where Rf and R ^g are each independently H, C 1-4 alkyl, -C(O)C1-4-alkyl, -C(O)OC1-4-alkyl or -S(O)2C1.4-alkyl. In some embodiments, R ¹ is -NR ^y R ^z , where one of R ^y and R ^z is H and the other is C1-4 alkyl unsubstituted or substituted with hydroxyl. In some embodiments, R ¹ is -NH(CH2)2OH.

In some embodiments, R ¹ is an optionally substituted C1-4 alkoxy group. In some embodiments, R ¹ is an unsubstituted _C1-4 alkoxy group. In other embodiments, R ¹ is C1-4 alkoxy substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NRfR ^g , cyano groups, nitro groups, _C1-4 alkoxy groups, _C1-4 haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C (O)NRfR ^g and -Oc(O)NRfR ^g where R ⁴ is H or C1-4 alkyl and Rf and R ^g are each independently H, C1-4 alkyl, -C(O)C ₁ - ₄ -alkyl, -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ¹ is a C1-4 alkoxy group further substituted with a C1-4 alkoxy group. For example, in some embodiments, R ¹ is -OCH2CH ₂ OCH ₂ CH ₃ or -OCH ₂ CH ₂ OCH ₃ . In other embodiments, R ¹ is a C1_4 alkoxy group substituted with an optionally substituted C1_4 alkoxy group. In some embodiments, R ¹ is -(OCH2CH ₂ ) _p -O-CH ₂ CH ₃ , where p is 0-10. In other embodiments, R ¹ is -(0CH ₂ CH ₂ ) _p -0-CH ₃ , where p is 0-10.

In some embodiments, R ² is a hydrogen atom, hydroxy, halogen, optionally substituted C1-4 alkyl, optionally substituted C1-4 alkoxy, or -NR ^x R ^y . In some embodiments, R ² is a hydrogen atom. In some embodiments, R ² is hydroxyl. In some embodiments, R ² is halogen. In some embodiments, R ² is chlorine. In some embodiments, R ² is fluoro. In other embodiments, R ² is bromine or iodine. In some embodiments, R ² is an optionally substituted C1-4 alkyl. In some embodiments, R ² is C1-4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NRfR ^g , cyano, nitro, _C1-4 alkoxy, and C ₁ - ₄ haloalkoxy groups wherein Rf and R ^g are each independently H, C1-4 alkyl, -C(O)C1-4 alkyl, -C(O)OC1-4 alkyl, or -S (O)2C1.4alkyl. In some embodiments, R ² is C1-4 alkyl substituted with one or more halogen atoms. In some embodiments, R ² is -CF ₃ , -(CH ₂ )F,

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-CHF2, CH ₂ Br, -CH ₂ CF ₃ , -CH2CHF2 or -CH2CH2F. In some embodiments, R ² is CF ₃ . In some embodiments, R ² is unsubstituted Ox-alkyl.

For example, in some embodiments, R ² is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, or t-butyl.

In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z , together with the nitrogen atom to which they are attached, form an optionally substituted monocyclic heterocycloalkyl ring. In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z together with the nitrogen atom to which they are attached form an optionally substituted 5-12 membered heterocycloalkyl ring. In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z , together with the nitrogen atom to which they are attached, form an optionally substituted 5-6 membered heterocycloalkyl ring. In some embodiments, R ² is morpholinyl. In some embodiments, R ² is morpholinyl substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NRfR ^s , cyano- groups, nitro groups, CM-alkoxy groups, C1. ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NRfR ^s and -OC(O)NRfR ^s where R ⁴ is is H or C1.4-alkyl; and Rf and ^{Rs are} each independently H, C1. ₄ -alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ . ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ² is piperazinyl. In some embodiments, R ² is piperazinyl substituted with one or more substituents selected from the group consisting of C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, hydroxyl, halo, -NR ^f R ^s , cyano groups, nitro groups, C1.4 alkoxy groups, C1.4 haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C1. ₄ -alkyl and R ^f and R ^s are each independently H, C ₁ . ₄ -alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ . ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ² is piperidinyl. In some embodiments, R ² is piperidinyl substituted with one or more substituents selected from the group consisting of C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, C1.4 alkoxy groups, C1.4 haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C1. ₄ -alkyl and R ^f and R ^s are each independently H, C ₁ . ₄ -alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ . ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ² is pyrrolidinyl. In some embodiments, R ² is pyrrolidinyl substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, C1. ₄ -alkoxy groups, C ₁ . ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^s and -OC(O)NR ^f R ^s , where R ⁴ is H or C1.4 alkyl and R ^f and R ^s are each independently H, C1. ₄ -alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ . ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl.

In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z are each independently H or optionally substituted C1.4 alkyl. In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z are each H. In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z are each optionally substituted C1. ₄ -alkyl. In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z are each optionally substituted C ₁ . ₄ -alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NR ^f R ^s , cyano, nitro, C1.4-alkoxy, C1.4-haloalkoxy, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NR ^f R ^s and -OC(O)NR ^f R ^s where R ⁴ is H or C1. ₄ alkyl and R ^f and R ^s are each independently H, C ₁ . ₄ -alkyl, -C(O)C ₁ . ₄ -alkyl, -C(O)OC ₁ . ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z are each optionally substituted C1.4 alkyl. In some embodiments, R ² is -N(CH2) ₂ or -N(CH ₂ CH ₃ ) ₂ . In some embodiments, R ² is -NR ^y R ^z , where R ^y and R ^z are each unsubstituted C1.4 alkyl or C1.4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, C1. ₄ alkoxy groups and C ₁ . ₄ -haloalkoxy groups, where R ^f and R ^s are each independently H, C1.4-alkyl, -C(O)C1.4-alkyl, -C(O)OC1.4-alkyl, or -S(O )2C1.4-alkyl. In some embodiments, R ² is -NR ^y R ^z , where one of R ^y and R ^z is H and the other is unsubstituted C1. ₄ -alkyl. In other embodiments, R ² is -NR ^y R ^z , where one of R ^y and R ^z is H and the other is C1.4-alkyl substituted with one or more substituents selected from the group consisting of hydroxyl , halogen, -NR ^f R ^s , cyano groups, nitro groups, C1. ₄ -alkoxy, and C ₁ . ₄ haloalkoxy groups, wherein R ^f and R ^s are each independently H, C1.4 alkyl, -C(O)C1.4 alkyl, -C(O)OC1.4 alkyl, or -S( O)2C1.4-alkyl. In some embodiments, R ² is -NR ^y R ^z , where one of R ^y and R ^z is H and the other is C1. ₄ -alkyl having no substituents or substituted with hydroxyl. In some embodiments, R ² is -NH(CH ₂ ) ₂ OH.

In some embodiments, R ² is an optionally substituted

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C1-4 alkoxy group. In some embodiments, R ² is an unsubstituted C1-4 alkoxy group. In other embodiments, R ² is C1-4 alkoxy substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halo, -NRfR ^s , cyano groups, nitro groups, _C1-4 alkoxy groups, _C1-4 haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NRfR ^s and -OC(O)NRfR ^s where R ⁴ is H or C 1-4 alkyl and Rf and R ^s are each independently H, C 1-4 alkyl, -C(O) C ₁ - ₄ -alkyl, -C (O) OC ₁ - ₄ -alkyl or -S (O) ₂ C ₁ _ ₄ -alkyl. In some embodiments, R ² is a C1-4 alkoxy group further substituted with a C1-4 alkoxy group. For example, in some embodiments, R ² is —OCH2CH ₂ OCH ₂ CH ₃ or —OCH ₂ CH ₂ OCH ₃ . In other embodiments, R ² is a C1-4 alkoxy group substituted with an optionally substituted C1.4 alkoxy group. In some embodiments, R ² is -(OCH2CH ₂ ) _p -O-CH ₂ CH ₃ , where p is 0-10. In other embodiments, R ² is -(OCH ₂ CH ₂ ) _p -O-CH ₃ , where p is 0-10.

In some embodiments, R ³ is a hydrogen atom, hydroxy, halogen, optionally substituted C1-4 alkyl, optionally substituted C1-4 alkoxy, or -NR ^x R ^y . In some instances, C1-4alkyl or _C1-4alkoxy groups are substituted _with one or more substituents selected from the group consisting of hydroxyl, halogen, -NR ^f R ^s , cyano, nitro, C1-4 -alkoxy groups and C1-4haloalkoxy groups, where Rf and R ^s each independently represent H, Ci-4-alkyl, -C(O) _C1-4 -alkyl, -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ _ ₄ -alkyl. In some embodiments, R ³ is a hydrogen atom. In some embodiments, R ³ is hydroxyl. In some embodiments, R ³ is halogen. In some embodiments, R ³ is chlorine. In some embodiments, R ³ is fluoro. In other embodiments, R ³ is bromine or iodine. In some embodiments, R ³ is an optionally substituted C1-4 alkyl. In some embodiments, R ³ is C 1-4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NR ^f R ^s , cyano, nitro, C 1-4 alkoxy groups and C1-4 haloalkoxy groups, where Rf and R ^s are each independently H, C1-4 alkyl, -C(O)C _{1 -4} alkyl, -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ³ is C1-4 alkyl substituted with one or more halogen atoms. In some embodiments, R ³ is -CF3, -(CH ₂ )F, -CHF ₂ , CH ₂ Br, -CH ₂ CF ₃ , -CH ₂ CHF ₂ , or -CH ₂ CH ₂ F. In some embodiments, R ³ is CF3. In some embodiments, R ³ is unsubstituted C1-4 alkyl. For example, in some embodiments, R ³ is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, or tert-butyl.

In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z , together with the nitrogen atom to which they are attached, form an optionally substituted monocyclic heterocycloalkyl ring. In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z together with the nitrogen atom to which they are attached form an optionally substituted 5-12 membered heterocycloalkyl ring. In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z , together with the nitrogen atom to which they are attached, form an optionally substituted 5-6 membered heterocycloalkyl ring. In some embodiments, R ³ is morpholinyl. In some embodiments, R ³ is morpholinyl substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NRfR ^s , cyano- groups, nitro groups ^{, C1-4 alkoxy groups, C1-4 haloalkoxy groups, -C(O)R 4 , -OC(O)R 4} _{, -C (} ^O )OR ⁴ , -C(O) NRfR ^s and -OC(O)NR ^f 'R ^s , where R ⁴ is H or C1-4 alkyl and Rf and R ^s are each independently H, C1-4 alkyl, -C(O)C ₁ - ₄ -alkyl, -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ³ is piperazinyl. In some embodiments, R ³ is piperazinyl substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, _C1-4 alkoxy groups, C _1-4 haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O )NR ^f R ^s and -OC(O)NR ^f 'R ^s , where R ⁴ is H or C1-4 alkyl and Rf and R ^s are each independently H, C1-4 alkyl, -C(O )C ₁ - ₄ -alkyl, -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ³ is piperidinyl. In some embodiments, R ³ is piperidinyl substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, _C1-4 alkoxy groups, C _1-4 haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O )NR ^f R ^s and -OC(O)NR ^f 'R ^s , where R ⁴ is H or C1-4 alkyl and Rf and R ^s are each independently H, Cm alkyl, -C(O)C1 - ₄ -alkyl, -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ³ is pyrrolidinyl. In some embodiments, R ³ is pyrrolidinyl substituted with one or more substituents selected from the group consisting of C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, hydroxyl, halogen, -NR ^f R ^s , cyano groups, nitro groups, C _1-4 alkoxy groups, C ₁ - ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NRfR ^g and

-OC(O)NRfR ^s , where R ⁴ is H or C _1-4 -alkyl and Rf and R ^g are each independently H, C _1-4 -alkyl, -C(O)C ₁ - ₄ -alkyl , -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ _ ₄ -alkyl.

In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z are each independently H or optionally substituted C1-4 alkyl. In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z are each H. In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z are each optionally substituted C1-4-alkyl. In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z are each optionally substituted C1-4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NRfR ^g , cyano groups, nitro groups, C1-4 alkoxy groups, C ₁ - ₄ haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NRfR ^g and -OC(O)NRfR ^g where R ⁴ is H or C1-4 alkyl and Rf and R ^g are each independently H, C1-4 alkyl, -C(O)C _{1 -4} alkyl, -C(O)OC ₁ - ₄ -alkyl or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z are each optionally substituted C1-4 alkyl. In some embodiments, R ³ is -N(CH2)2 or -N(CH2CH ₃ ) ₂ . In some embodiments, R ³ is -NR ^y R ^z , where R ^y and R ^z are unsubstituted C1-4 alkyl or C1-4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen , -NR ^f R ^g , cyano groups, nitro groups, _C1-4 alkoxy groups and _C1-4 haloalkoxy groups, wherein Rf and R ^g are each independently H, C1-4 alkyl, -C(O )C1-4-alkyl, -C(O)OC1-4-alkyl or -S(O)2C1_4-alkyl. In some embodiments, R ³ is -NR ^y R ^z , where one of R ^y and R ^z is H and the other is unsubstituted C1-4 alkyl. In other embodiments, R ³ is -NR ^y R ^z , where one of R ^y and R ^z is H and the other is C1-4 alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen, -NR ^f R ^g , cyano groups, nitro groups, C 1-4 alkoxy groups and C 1_ ₄ haloalkoxy groups, where Rf and R ^g are each independently H, C 1-4 alkyl, -C (O)C1-4-alkyl, -C(O)OC1-4-alkyl or -S(O)2C1_4-alkyl. In some embodiments, R ³ is -NR ^y R ^z , where one of R ^y and R ^z is H and the other is C1-4 alkyl, unsubstituted or substituted with hydroxyl. In some embodiments, R ³ is -NH(CH ₂ ) ₂ OH.

In some embodiments, R ³ is an optionally substituted C1-4 alkoxy group. In some embodiments, R ³ is an unsubstituted _C1-4 alkoxy group. In other embodiments, R ³ is C1-4 alkoxy substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxyl, halo, -NRfR ^g , cyano groups, nitro groups, Cm alkoxy groups, C1-4 haloalkoxy groups, -C(O)R ⁴ , -OC(O)R ⁴ , -C(O)OR ⁴ , -C(O)NRfR ^g and -OC(O)NRfR ^g where R ⁴ is H or C1-4 alkyl and Rf and R ^g are each independently H, Ci-4 alkyl, -C(O) _C1-4 alkyl, -C(O)OC _{1 -4} alkyl, or -S(O) ₂ C ₁ . ₄ -alkyl. In some embodiments, R ³ is a C1-4 alkoxy group further substituted with a C1-4 alkoxy group. For example, in some embodiments, R ³ is —OCH2CH ₂ OCH ₂ CH ₃ or —OCH ₂ CH ₂ OCH ₃ . In other embodiments, R ³ is a C1-4 alkoxy group substituted with an optionally substituted C1_4 alkoxy group. In some embodiments, R ³ is -(OCH2CH ₂ ) _p -O-CH ₂ CH ₃ , where p is 0-10. In other embodiments, R ³ is -(OCH ₂ CH ₂ ) _p -O-CH ₃ , where p is 0-10.

In some embodiments, R ¹ , R ² and R ³ are independently selected from the group consisting of H, -C1, -CN, -CF3, methyl, methoxy, -MHCH2CH2OH, -H(CH2CH3)2, -N(CH3 )2, -OCH2CH2-O-CH2CH3, -OCH2CH2OCH3, morpholinyl, 4-methyl-piperazin-1-yl, piperidinyl and pyrrolidinyl. In some embodiments, R ¹ is selected from the group consisting of H, -INSH2CH2OH, -N(CH2CH ₃ ) ₂ , morpholinyl, 4-methyl-piperazin-1-yl, piperidinyl, pyrrolidinyl, -OCH2CH2-O-CH2CH3 and OCH ₂ CH ₂ OCH ₃ . In some embodiments, R ² is selected from the group consisting of H, -CF ₃ , -CN, methyl, methoxy, -OCH ₂ CH ₂ -O-CH ₂ CH ₃ , -OCH ₂ CH ₂ OCH ₃ , -N (CH ₃ ) ₂ and morpholinyl. In some embodiments, R ³ is selected from the group consisting of H, -C1, -CN, methyl, methoxy, and morpholinyl.

It should be understood that the definitions of any variable in formula (I) may, where applicable, be combined with one or more definitions of any other variable, as if each and every combination of variables were specifically and separately listed. For example, each value of R ¹ can be combined with each value of R ² and R ³ as if any combination thereof were specifically and separately listed. Similarly, each R ² value may be combined with each R ¹ and R ³ value as if any combination thereof were specifically and separately listed, and each R ³ value may be combined with each R ¹ and R ² value as if any combination of them has been specially and separately listed.

In some embodiments, a compound having formula (I) is a compound shown in the following table.

- 16 042432

Compound Structure Name 1 “P q o Chya 0'“ hh ζ^ ly ΖΞΓ 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione 2 Οχ ,0 0000. 0 4-(4-((4-chloro-3- (trifluoromethyl)phenyl)sulfonyl)-2morpholinophenyl)-2,4-dihydro-3H-1,2,4- triazole-3-thione 3 Οχ,о ash I , ^NH 4-(4-(phenylsulfonyl)phenyl)-2,4-dihydro-3H- 1,2,4-triazole-3-thione 4 t z 0Vz .0% O 4-(4-((4-chlorophenyl)sulfonyl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione 5 TJ T1 J _C| /X/ 4-(4-((4-chloro-3-methylphenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione 6 z about 0. ωζ 0' z I 4-((4-(5-thioxo-1,5-dihydro-4H-1,2,4-triazole- 4-yl)phenyl)sulfonyl)benzonitrile

- 17 042432

7 0 0. ωζ 0° ζ^ω ζ τ 4-(4-((4-morpholinophenyl)sulfonyl)phenyl)- 2,4-dihydro-3H-1,2,4-triazole-3-thione 8 / Ο X. 0'° ζ I 4-(4-((4-Methoxyphenyl)sulfonyl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione 9 0 0 w. „ 4-(4-tosylphenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione 10 XX XX L _F ^Z 4-(4-((4-fluorophenyl)sulfonyl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione eleven τ Ζ ₄ wv π .0 ;ω X ο LL 4-(4-((3-(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione 12 \ ο 0.0 ω; 0'° ζΛ« ζ I 4-(4-((3-methoxyphenyl)sulfonyl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione 13 ο ο 0 Η Ί Η Ί θ Ο l ^^νΑη ^Ν 4-(4-((3-(2-ethoxyethoxy)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione

- 18 042432

14 z about 7,. ωί 0 ζ τ 3 -((4-(5 -thioxo-1,5 -dihydro-4H-1,2,4-triazole- 4-yl)phenyl)sulfonyl)benzonitrile 15 τ z Ος-/ζ „0 5ω ζ -ζ \ 4-(4-((3-(dimethylamino)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione 16 °χ/° XX ΥΥ j ^νΑ _νη 4-(4-((3-morpholinophenyl)sulfonyl)phenyl)- 2,4-dihydro-3H-1,2,4-triazole-3-thione 17 SL' 0' £>» Τ 4-(4-(m-tolylsulfonyl)phenyl)-2,4-dihydro3H-1,2,4-triazole-3-thione 18 °\/° 0¾ HN. L= _n ' ΌΗ 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-((2-hydroxyethyl)amino)phenyl)- 2,4-dihydro-3H-1,2,4-triazole-3-thione 19 0 _χ 0 “Wz ο 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-(piperidin-1-yl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione 20 0 _χ 0 X%M1 0 1 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-(4-methylpiperazin-1-yl)phenyl)- 2,4-dihydro-3H-1,2,4-triazole-3-thione

- 19 042432

4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-(diethylamino)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione

4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfony l) -2 -(2-ethoxyethoxy)phenyl) -2,4 dihydro-3H-1,2,4-triazole-3 -tion

4-(4-((4-methyl-3-(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione

4-(4-((4-Chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-(pyrrolidin-1-yl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione

4-(4-((3-(2-Methoxyethoxy)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione or a pharmaceutically acceptable salt thereof.

Compounds having formula (I) may be prepared and/or formulated as pharmaceutically acceptable salts. In some embodiments, pharmaceutically acceptable salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or they may be formed with organic acids such as acetic acid, oxalic acid, propionic acid, succinic acid, maleic acid, tartaric acid, and the like. These salts can be derived from inorganic or organic acids. Non-limiting examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-1,4-dioates, hexine-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulfonates, methylsulfonates, propylsulfonates, besylates, xylenesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, γ-hydroxybutyrates, glycolates, tartrates and mandelates. In some embodiments, pharmaceutically acceptable salts are formed when an acidic proton present in the parent compound is either replaced by a metal ion, such as an alkali metal ion, an alkaline earth metal ion, or an aluminum ion, or is coordinated to an organic base. Salts formed with pharmaceutically acceptable organic non-toxic bases include salts with primary, secondary and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2diethylaminoethanol, tromethamine, trimetamine, dicyclohexylamine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-ethylglucamine, N-methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, amino acids such as lysine , arginine, histidine, etc. Examples of pharmaceutically acceptable salts with bases include salts with inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper,

- 20 042432 manganese, aluminum, etc. In some embodiments, the organic non-toxic bases are L-amino acids such as L-lysine and L-arginine, tromethamine, N-ethylglucamine, and N-methylglucamine. Suitable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like. Lists of other suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th edition, Mack Publishing Company, Easton, Pa., 1985.

For a compound containing a basic nitrogen atom described herein, a pharmaceutically acceptable salt can be prepared by any suitable method available in the art, for example by reacting the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, nitric acid, boric acid, phosphoric acid and the like, or with an organic acid such as acetic acid, phenylacetic acid, propionic acid, stearic acid, lactic acid, ascorbic acid, maleic acid, hydroxymaleic acid, isethionic acid, succinic acid , valeric acid, fumaric acid, malonic acid, tartaric acid, oxalic acid, glycolic acid, salicylic acid, oleic acid, palmitic acid, lauric acid, pyranosidyl acid such as glucuronic acid or galacturonic acid, alpha hydroxy acid such as mandelic acid , citric acid or tartaric acid, an amino acid such as aspragic acid or glutamic acid, an aromatic acid such as benzoic acid, 2-acetoxybenzoic acid, naphthoic acid or cinnamic acid, a sulfonic acid such as lauryl sulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid or ethanesulfonic acid , or any compatible mixture of acids, such as described in the examples in this text, and any other acid and mixtures thereof that can be considered equivalents or acceptable replacements in light of the standard level of knowledge in this field.

Embodiments also relate to pharmaceutically acceptable prodrugs of the compounds described herein and methods of treatment using such pharmaceutically acceptable prodrugs. The term prodrug means a precursor of said compound which, after administration to a patient, yields the target compound in vivo by a chemical or physiological process such as solvolysis or enzymatic cleavage, or under physiological conditions (e.g., a prodrug at physiological pH is converted to a compound having the formula (I )). A pharmaceutically acceptable prodrug is a prodrug that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject. Illustrative procedures for selecting and preparing suitable prodrugs are described, for example, in Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.

Embodiments also relate to pharmaceutically active metabolites of the compounds described herein and the use of such metabolites in the methods described herein. A pharmaceutically active metabolite means a pharmaceutically active product of the metabolism of a compound described herein or a salt thereof in the body. Prodrugs and active metabolites of a compound can be determined using standard techniques known in the art. See, for example, Bertolini et al, J. Med. Chem., 1997, 40, 2011-2016; Shan et al., J. Pharm. Sci., 1997, 86(7), 765-767; Bagshawe, Drug Dev. Res., 1995, 34, 220-230; Bodor, Adv. Drug Res., 1984, 73, 255-331; Bundgaard, Design of Prodrugs (Elsevier Press, 1985); and Larsen, Design and Application of Prodrugs, Drug Design and Development (Krogsgaard-Larsen et al., eds., Harwood Academic Publishers, 1991).

pharmaceutical compositions.

When used for treatment, the pharmaceutical composition of the present invention contains at least one compound having formula (I) or a pharmaceutically acceptable salt thereof. The pharmaceutical compositions may further contain one or more pharmaceutically acceptable excipients. A pharmaceutically acceptable excipient is one that is non-toxic and otherwise biologically suitable for administration to a subject. Such excipients facilitate the administration of the compounds described in this text and are compatible with the active substance. Examples of pharmaceutically acceptable excipients include stabilizers, lubricants, surfactants, diluents, antioxidants, binders, colorants, bulking agents, emulsifiers, or flavor modifiers. In some embodiments, the pharmaceutical compositions of the present invention are sterile compositions. Pharmaceutical compositions can be prepared according to known pharmaceutical formulation techniques, as well as those available to those skilled in the art.

Sterile compositions are also covered by embodiments of the present invention, including compositions that comply with national and local regulatory requirements for such compositions.

The pharmaceutical compositions and compounds described herein may be in the form of formulations such as solutions, emulsions, suspensions, dispersions or inclusion complexes such as cyclodextrins in suitable pharmaceutical solvents or carriers, or in the form of pills, tablets

- 21 042432 current, cast tablets, suppositories, sachets, dragees, granules, powders, powders for dissolution before administration or capsules, with solid carriers, in accordance with conventional methods known in the field of preparation of various dosage forms. The pharmaceutical compositions described herein may be administered by a suitable route of administration such as oral, parenteral, rectal, nasal, topical or intraocular, as well as by inhalation. In some embodiments, the compositions are in the form of intravenous or oral preparations.

For oral administration, the compounds of the present invention may be presented in solid form such as a tablet or capsule, or in the form of a solution, emulsion or suspension. For the preparation of compositions for oral administration, the compounds described herein may be formulated at a dosage of, for example, from about 0.01 mg/kg to about 50 mg/kg per day, or from about 0.05 mg/kg to about 20 mg/kg per day. day, or from about 0.1 mg/kg to about 10 mg/kg per day. Tablets for oral administration may contain the active ingredient(s) in admixture with compatible pharmaceutically acceptable excipients such as diluents, disintegrants, binders, lubricants, sweeteners, flavors, colors and preservatives. Suitable excipients include sodium and calcium carbonate, sodium and calcium phosphate, lactose, starch, sugar, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol, and the like. Examples of liquid excipients for oral formulations include ethanol, glycerin, water, and the like. Starch, polyvinyl pyrrolidone (PVP), sodium starch glycolate, microcrystalline cellulose and alginic acid are examples of disintegrants. Binders may include starch and gelatin. Lubricants, if present, may be magnesium stearate, stearic acid or talc. If desired, tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract, or may be coated with an enteric coating.

Capsules for oral administration include hard and soft gelatin capsules. To obtain hard gelatin capsules, the active ingredient(s) are mixed with a solid, semi-solid or liquid diluent. Soft gelatin capsules can be prepared by mixing the active ingredient with water, an oil such as peanut oil or olive oil, liquid paraffin, a mixture of mono- and diglycerides of short chain fatty acids, polyethylene glycol 400 or propylene glycol.

Oral liquids may be in the form of suspensions, solutions, emulsions or syrups, or they may be lyophilized and presented as a dry product for reconstitution in water or other suitable vehicle prior to use. Such liquid compositions may optionally contain pharmaceutically acceptable excipients such as suspending agents (eg, sorbitol, methylcellulose, sodium alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel, etc.);

non-aqueous vehicles such as oil (eg almond oil or fractionated coconut oil), propylene glycol, ethyl alcohol or water;

preservatives (eg methyl or propyl parahydroxybenzoate or sorbic acid);

wetting agents such as lecithin; and optionally flavorings or colorings.

The compositions described herein may take the form of rectal preparations such as suppositories. For parenteral use, including intravenous, intramuscular, intraperitoneal, intranasal or subcutaneous, the compounds described herein can be administered in sterile aqueous solutions or suspensions buffered to an appropriate pH and isotonicity, or in oils suitable for parenteral administration. Suitable aqueous vehicles include Ringer's solution and isotonic sodium chloride solution. Such forms may be presented in single dose form such as ampoules or disposable injection devices, in forms containing multiple single doses such as vials from which the required portions can be taken, or in solid or concentrate form which can be used for preparation of an injectable. Exemplary infusion dosages are in the range of about 1 to 1000 μg/kg/min of the substance in admixture with a pharmaceutical carrier over a period of several minutes to several days.

For nasal, inhalation or oral administration, the compounds or pharmaceutical compositions described herein can be administered using, for example, a spray also containing a suitable carrier.

In some topical embodiments, the compounds of the present invention are formulated in creams, ointments, or similar vehicles suitable for topical use. For topical use, the compounds or pharmaceutical compositions described herein may be mixed with a pharmaceutical carrier at a concentration of from about 0.1% to about 10% drug in vehicle. In another route of administration, those described herein

- 22 042432 the text of the substance can be introduced into the composition of the drug, in the form of a patch, to achieve transdermal administration.

Preparations obtained by the method of dry spraying.

In some embodiments, the present invention describes pharmaceutical preparations containing compounds having formula (I) that optimize the bioavailability of the compounds. In some embodiments, the pharmaceutical preparations are in the form of an amorphous dispersion. In some embodiments, pharmaceutical formulations are dry sprayed to form dry spray dispersions (SDD). Dry spraying is a process in which a compound and excipients are dissolved in a common solvent and the resulting solution is sprayed into a drying chamber. During this process, the liquid solution containing the active compound is converted into fine dry particles.

In some embodiments, dry spraying involves contacting a liquid suspension or solution containing one or more compounds having formula (I) and one or more pharmaceutically acceptable excipients with a sufficient volume of hot air to evaporate and dry the liquid droplets. The dry spray formulation can be any solution, suspension, slurry, colloidal dispersion, or paste that can be finely sprayed using the dry spray apparatus of choice. In some embodiments, the liquid slurry is sprayed into a stream of warm, filtered air that vaporizes the solvent and carries the dry product to a collector (eg, cyclone). The exhaust air is then stripped of the solvent, or alternatively the exhaust air is sent to a cooler to capture and potentially recycle the solvent. Commercially available types of apparatus can be used to carry out dry spraying.

In some embodiments, the dry spray formulation contains from about 3% to about 40% by weight of a compound of the present invention, for example, from about 3% to about 35%, from about 3% to about 30%, from about 3 % to about 25%, from about 3% to about 20%, from about 3% to about 15%, from about 3% to about 10%, from about 3% to about 5%, from about 10% to about 35% , from about 10% to about 30%, from about 10% to about 25%, from about 10% to about 20%, from about 15% to about 35%, from about 15% to about 30%, from about 15% up to about 25%, from about 15% to about 20%, from about 20% to about 35%, from about 20% to about 30%, from about 20% to about 25%, from about 25% to about 35%, from about 25% to about 30%, or from about 30% to about 40% by weight. In some embodiments, the dry spray formulation contains about 5% by weight of a compound of the present invention. In some embodiments, the dry spray formulation contains about 10% by weight of a compound of the present invention. In some embodiments, the dry spray formulation contains about 15% by weight of a compound of the present invention. In some embodiments, the dry spray dispersion contains at least about 10% by weight of a compound of the present invention. In general, the upper limit of the solids content is determined by the viscosity (eg, pumpability) of the resulting solution and the solubility of the components in the solution. In general, the viscosity of the solution can determine the particle size of the resulting powdered product.

In some embodiments, the dry spray dispersion contains one or more compounds having formula (I) and one or more pharmaceutically acceptable excipients. In some embodiments, one or more pharmaceutically acceptable excipients include one or more binders. In some embodiments, the binder is polymeric. In some embodiments, the one or more binders are selected from the group consisting of polymeric cellulose derivatives such as carboxymethyl cellulose (CMC), hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC), and hydroxypropyl methyl cellulose (HPMC); gelatin; gelatin hydrolyzate; sucrose; dextrose; and non-cellulose binders such as polyvinylpyrrolidone (PVP), polyvinylpyrrolidone/vinyl acetate (PVP/VA), polyethylene glycol (PEG), vinylpyrrolidone copolymers, pregelatinized starch, sorbitol and glucose; methacrylate and polyacrylates. In some embodiments, one or more binders are selected from the group consisting of polyvinylpyrrolidone and its derivatives, such as collidone; cellulose derivatives such as HPMC; and polyoxyethylene/polyethylene glycol polymers such as PEG. In some embodiments, one or more binders are selected from the group consisting of PVP, PVP/VA, and HPMC. In some embodiments, one or more binders are selected from the group consisting of PVP-VA 64, HPMC E5, HPMC-AS, and collidone 30. In some embodiments, the binder includes PVP-VA 64. In some embodiments, the binder includes collidone 30. In some embodiments, the binder comprises HMPC E5. In some embodiments, the binder comprises HMPC-AS. In some embodiments, the dry spray dispersion contains at least one polymer

- 23 042432 and at least one compound having the formula (I). In some embodiments, the dry spray dispersion contains a polymer and a compound of the present invention in a ratio of about 10:1 to about 1:1, such as about 5:1 to about 2:1.

In some embodiments, dry spray solutions contain a polymer and a compound of the present invention in a ratio of about 3:1.

In some embodiments, dry spraying is carried out at an inlet temperature of from about 60°C to about 200°C, for example, from about 95°C to about 185°C, from about 110°C to about 182°C, or from about 96°C. up to about 180°C. In some embodiments, dry spraying is carried out at an inlet temperature of about 145°C. In some embodiments, dry spraying is carried out at an outlet temperature of from about 30°C to about 90°C, for example, from about 30°C to about 80°C, from about 30°C to about 70°C, from about 30°C up to about 60°C or from about 30°C to about 50°C. In some embodiments, the implementation of dry spraying is carried out at an outlet temperature of from about 35°C to about 45°C. In some embodiments, dry spraying is carried out at an outlet temperature of about 40°C. In some embodiments, the spray flow rate is from about 1 g/min to about 50 g/min, for example, from about 1 g/min to about 40 g/min, from about 1 g/min to about 30 g/min, from about 1 g/min to about 20 g/min, about 1 g/min to about 10 g/min, about 5 g/min to about 40 g/min, about 5 g/min to about 30 g/min , from about 5 g/min to about 20 g/min, from about 5 g/min to about 10 g/min, from about 10 g/min to about 40 g/min, from about 10 g/min to about 30 g /min, from about 10 g/min to about 20 g/min, from about 20 g/min to about 40 g/min, from about 20 g/min to about 30 g/min, from about 30 g/min to about 40 g/min, or from about 40 g/min to about 50 g/min. In some embodiments, the spray flow rate is from about 5 g/min to about 15 g/min, such as about 8 g/min or about 10 g/min.

In some embodiments, removal of the solvent may require a subsequent drying step, such as tray drying, fluid bed drying (e.g., from about room temperature to about 100°C), vacuum drying, microwave drying, rotary drum drying, or biconical vacuum drying. drying (for example, at a temperature from about room temperature to about 200°C). In one embodiment, the solid dispersion is dried in a fluidized bed.

In one embodiment of the dry spray process, the solvent includes a volatile solvent, such as a solvent having a boiling point of less than about 100°C. In some embodiments, the solvent is a mixture of solvents, such as a mixture of volatile solvents or a mixture of volatile and non-volatile solvents. When a mixture of solvents is used, the mixture may include one or more non-volatile solvents, for example, when the non-volatile solvent is present in the mixture in an amount of less than about 15%, such as less than about 12%, less than about 10%, less than about 8%, less than about 5%. , less than about 3% or less than about 2%.

In some embodiments, compounds having formula (I) have a solubility of at least about 10 mg/mL (e.g., at least about 15, 20, 25, 30, 35, 40, 45, 50 mg/mL, or greater) in the solvent used for the dry spray procedure. In some embodiments, compounds having formula (I) have a solubility of at least about 20 mg/mL in the solvent used for the dry spray procedure.

Examples of solvents that can be used for the dry spray procedure include acetone, cyclohexane, dichloromethane (DCM), K,K-dimethylacetamide (DMA), K,K-dimethylformamide (DMF), 1,3-dimethyl-2-imidazolidinone (DMI ), dimethyl sulfoxide (DMSO), dioxane, ethyl acetate, diethyl ether, glacial acetic acid (HAc), methyl ethyl ketone (MEK), N-methyl-2-pyrrolidinone (NMP), methyl tert-butyl ether (MTBE), tetrahydrofuran (THF ), pentane, acetonitrile, methanol, ethanol, isopropyl alcohol, isopropyl acetate, toluene and water. Examples of co-solvents include acetone/THF, acetone/methanol, acetone/ethanol, acetone/ethyl acetate, acetone/DCM, acetone/DMSO, acetone/DMF, acetone/water, ethyl acetate/DCM, ethyl acetate/THF, ethyl acetate/methanol, ethyl acetate/ethanol , MEK/water, THF/water, THF/methanol, THF/ethanol, dioxane/water, DCM/methanol, DCM/ethanol, and DCM/THF. In dual solvent systems, the solvents may be present in amounts from about 0.1% to about 99.9%. In some embodiments, DXM is used as a co-solvent with methanol in a ratio of about 90:10 to about 60:40, such as about 80:20. In some embodiments, the solvent system contains three solvents. For example, acetone and water can be mixed with a third solvent such as DMA, DMF, DMI, DMSO, or HA. In some embodiments, the solvents used in the dry spray procedure dissolve both the compound and the polymer.

In some embodiments, dry spray (SDD) dispersions contain from about 10% to about 75% by weight of a compound of the present invention, such as from about 10% to about 65%, from about 10% to about 55% from about 10% to about 45%, from about 10% to about 35%, from about 10% to about 25%, from about 10% to about 15%, from about 25% to about 75%, from about 25% to about 65%, from about 25% to about 55%, from about 25% to about 45%, from about 25% to about 35%, from about 35% to about 75%, from about 35% to about 65%, from about 35% to about 55%, from about 35% to about 45%, from about 45% to about 75%, from about 45% to about 65%, from about 45% to about 55%, from about 55% to about 75%, or from about 65% to about 75% by weight. In some embodiments, the dry spray formulation contains about 25% by weight of a compound of the present invention. In some embodiments, the dry spray formulation contains about 30% by weight of a compound of the present invention. In some embodiments, the dry spray formulation contains about 50% by weight of a compound of the present invention. In some embodiments, the dry spray dispersion contains at least about 20% by weight of a compound of the present invention.

In some embodiments, dry spray dispersions (SDDs) are included in the final dosage form. Examples of final dosage forms include, but are not limited to, capsules, tablets, and sachets.

When used in this text, the terms treat or treatment means an approach to obtain beneficial or desired results, including clinical results. With respect to the compositions and methods described herein, beneficial or desirable clinical results include, but are not limited to, one or more of the following: relief of one or more symptoms resulting from the condition, reduction in the severity of the condition, stabilization of the condition ( for example, preventing or slowing the deterioration of a condition), alleviating a disease state, achieving remission of a disease (partial or complete), reducing the dosage of one or more other drugs needed to treat a condition, enhancing the effect of another drug used to treat a condition, increasing the quality of life of a patient with a pathological condition and / or an increase in survival time. A method for treating a disease or condition encompasses reducing the pathological consequences of the disease or condition. The methods described herein encompass any one or more of these aspects of treatment.

As used herein, the term prevent or prevent a condition, disease, or disorder means, in one embodiment, delaying or avoiding the onset of the disease or disorder (i.e., slowing or preventing the onset of the disease or disorder in a patient susceptible to developing the disease or condition). In some embodiments, the term prevent or prevention means delaying or slowing down the development of a pathological condition, disease, or disorder.

The term subject or patient means a mammalian patient in need of such treatment, such as a human.

Examples of diseases that may be therapeutic targets for such compounds include, but are not limited to, central neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease and other central neurodegenerative diseases and peripheral neurodegenerative diseases in which accumulation of neurotoxic proteins has been proven. .

In one aspect, the compounds and pharmaceutical compositions of the present invention specifically affect the accumulation of neurotoxic proteins or their aggregates. Thus, these compounds and pharmaceutical compositions can treat degenerative neurological diseases associated with or caused by dysregulation of protein homeostasis (proteostasis), for example, improper clearance of protein aggregates and/or damaged organelles, insufficient activation of a survival pattern of gene expression, and/or lack of energy drinks. cells. In some embodiments, the methods of the present invention target neurodegenerative diseases associated with the accumulation of neurotoxic misfolded and aggregated proteins. In some embodiments, the methods of treatment of the present invention target Parkinson's disease, Alzheimer's disease, Lewy body disease, multiple system atrophy, or Huntington's disease. The compounds, compositions and methods of the present invention are also useful in mitigating the detrimental effects of disturbed protein homeostasis, including disruption of various forms of macroautophagy and other protein clearance mechanisms. Although the present invention is not limited to any particular mechanism of action, autophagy dysregulation is believed to be caused by alpha-synuclein and beta-amyloid, as well as other proteins that accumulate and aggregate in neurodegenerative diseases.

In the methods of treatment of the present invention, the term effective amount means an amount or dose sufficient to achieve the desired therapeutic effect in subjects in need of such treatment. Effective amounts or dosages of the compounds described herein

- 25 042432 can be determined by standard methods, such as modeling, dose escalation or clinical studies, taking into account routine factors, for example, the method or route of administration of the drug, the pharmacokinetics of the drug, the severity and course of the infection, the general health of the patient, his condition, weight and opinion of the attending physician. Exemplary dosages are in the range of about 1 µg to 2 mg of active ingredient per kilogram of patient body weight, for example about 0.05 to 100 mg/kg/day, or about 1 to 35 mg/kg/day, or about 0.1 to 10 mg/kg/day. The total dosage can be given as a single or multiple divided doses (eg 2, 3 or 4 times a day).

Once there has been an improvement in the patient's health condition in the disease, the dosage may be changed to an appropriate prophylactic or maintenance treatment. For example, the dosage or frequency of administration, or both, may be reduced according to symptoms to a level that maintains the desired therapeutic or prophylactic effect. Of course, if the symptoms have been reduced to the desired level, treatment can be stopped. However, patients may require long-term intermittent therapy for any recurrence of symptoms.

Combinations of drugs.

The compounds described herein can be used in pharmaceutical compositions or methods in combination with one or more additional active ingredients in the treatment of neurogenerative diseases. For example, additional active ingredients are those that are known or will be discovered to be effective in the treatment of neurodegenerative diseases, including those that are active against another target associated with the disease, such as (but not limited to)

a) compounds that combat protein misfolding (such as drugs that decrease the production of such proteins, increase their clearance, or alter their aggregation and/or reproduction);

b) compounds that treat the symptoms of such disorders (eg, dopamine replacement therapy, cholinesterase inhibitors, and precognitive glutamatergic drugs); And

c) drugs that work as neuroprotectives by complementary mechanisms (eg those that act on autophagy, are antioxidants and work by other mechanisms, such as adenosine A2A antagonists).

For example, additional active ingredients are those known to be effective in the treatment of neurodegenerative diseases, including those active against another target associated with the disease, such as (but not limited to)

a) compounds that act on various protein misfolding mechanisms (such as aggregation and/or propagation);

b) compounds that treat the symptoms of such disorders (eg dopamine replacement therapy); And

c) drugs that act as neuroprotective agents by complementary mechanisms (eg, agents that act on autophagy, antioxidants, and adenosine A2A antagonists).

For example, the compositions, formulations, and treatments described herein may additionally include other drugs or pharmaceuticals, such as other active substances that can be used to treat or alleviate a degenerative neurological disease associated with or caused by protein aggregation, for example, synuclein aggregation. , beta-amyloid, tau, huntingtin or TDP43 protein, for example, Parkinson's disease, Alzheimer's disease, Lewy body disease and multiple system atrophy, or associated symptoms or pathological conditions. In this regard, the compositions and preparations of the compounds described herein can be used in methods of treating Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Lewy body dementia, dementia due to Parkinson's disease, multiple system atrophy, Huntington's disease, amyotrophic lateral sclerosis, cancer, inflammation, Crohn's disease, heart disease, aging, or traumatic brain injury (TBI). The pharmaceutical compositions described herein may further comprise one or more of such active ingredients, and the methods of treatment may further comprise administering an effective amount of one or more of such active ingredients. In some embodiments, the one or more additional active ingredients is a compound that is used to treat the symptoms or development of a neurodegenerative disease (eg, Alzheimer's disease, Parkinson's disease, Huntington's disease). In some embodiments, additional active agents may be cytokines, immunoregulatory agents, anti-inflammatory agents, complement activating agents such as peptides or proteins containing collagen-like domains or fibrinogen-like domains (e.g., ficolin), carbohydrate-binding domains, and the like. , as well as their combinations. In some embodiments, the additional active ingredient is an anti

- 26 042432 inflammatory agent. Additional active ingredients include compounds that may be used in such compositions and methods, including dopamine therapy drugs, catechin O-methyltransferase (COMT) inhibitors, monoamine oxidase inhibitors, cognitive impairment correctors (such as acetylcholinesterase inhibitors or memantine), adenosine 2A antagonists receptor, beta-secretase inhibitors or gamma-secretase inhibitors. In particular embodiments, at least one compound of the present invention may be combined in a pharmaceutical composition or method of treatment with one or more drugs selected from the group consisting of tacrine (Cognex), donepezil (Aricept), rivastigmine (Exelon), galantamine (Reminyl), physostigmine, neostigmine, icopezil (CP-118954, 5,7-dihydro-3-[2-[1(phenylmethyl)-4-piperidinyl]ethyl]-6H-pyrrolo-[4,5-f-] -1,2-benzisoxazol-6-one maleate), ER-127528 (4-[(5,6-dimethoxy-2-fluoro-1-indanone)-2-yl]methyl-1-(3-fluorobenzyl)piperidine hydrochloride), zanapezil (TAK-147; 3-[1 -(phenylmethyl)piperidin-4-yl]-1-(2,3,4,5-tetrahydro-1H-1-benzazepin-8-yl)-1- propane fumarate), metrifonate (T-588; (-)-R-alpha-[[2-(dimethylamino)ethoxy]methyl]benzo[b]thiophene-5-methanol hydrochloride), FK-960 (N-(4- acetyl-1-piperazinyl)-p-fluorobenzamide hydrate), TSN-346 (N-methyl-N-2-propynyldibenz[b,f]oxepin-10-methanamine), SDZ-220-581 ((S)-alpha-amino -5-(phosphonomethyl)-[1,1'biphenyl]-3-propionic acid), memantine (Namenda/Exiba) and 1,3,3,5,5-pentamethylcyclohexane-1amine (Neramexane), tarenflurbil (Flurizan), tramiprosate (Alzhemed), clioquinol, PBT-2 (an 8-hydroxyquinoline derivative), 1-(2-(2-naphthyl)ethyl)-4-(3-trifluoromethylphenyl)-1,2,3,6-tetrahydropyridine, Huperzine A, Posatirelin, Levprolide or its derivatives, Ispronicline, (3-Aminopropyl)(nbutyl )phosphinic acid (SGS-742), N-methyl-5-(3-(5-isopropoxypyridinyl))-4-penten-2-amine (ispronicline), 1-decanaminium, N-(2-hydroxy-3-sulfopropyl )-N-methyl-N-octyl, internal salt (zt-1), salicylates, aspirin, amoxypyrine, benorylate, cholinemagnesium salicylate, diflunisal, fislamin, methyl salicylate, magnesium salicylate, salicyl salicylate, diclofenac, aceclofenac, acemethacin, bromfenac, etodolac , indomethacin, nabumetone, sulindac, tolmetin, ibuprofen, carprofen, fenbufen, fenoprofen, flurbiprofen, ketoprofen, ketorolac, loxoprofen, naproxen, thiaprofenic acid, suprofen, mefenamic acid, meclofenam oic acid, phenylbutazone, azapropazone, metamizole, oxyphenbutazone, sulfinprazone, piroxicam, lornoxicam, meloxicam, tenoxicam, celecoxib, etoricoxib, lumiracoxib, parecoxib, rofecoxib, valdecoxib, nimesulide, arylalkanoic acids, 2-arylpropionic acids (profenyls), N-aryls (fenamic acids), pyrazolidine derivatives, oxicams, COX-2 inhibitors, sulfonanilides, essential fatty acids and minozac (2-(4-(4-methyl-6-phenylpyridazin-3-yl)piperazin-1yl)pyrimidine dihydrochloride hydrate). Such a combination may increase efficacy, relieve symptoms of other diseases, reduce one or more side effects, or reduce the required dosage of the compounds or compositions described herein. Additional active ingredients may be administered in a pharmaceutical composition separate from the compound of the present invention, or they may be included together with the compound of the present invention in the same pharmaceutical composition. Additional active ingredients can be administered simultaneously, before or after administration of the compound having formula (I).

Methods of treatment.

The present invention describes methods of treating a pathological condition associated with neurodegeneration or protein aggregation/accumulation, which include administering to a subject in need of such treatment, an effective amount of a compound or composition described in the present invention. Any of the compounds or pharmaceutical compositions described herein can be used in the treatment of a pathological condition associated with neurodegeneration or protein aggregation/accumulation. In some embodiments, the protein is alphasynuclein, a-beta, tau, huntingtin, or TDP43. In some embodiments, said condition is Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Lewy body dementia, Parkinson's disease dementia, multiple system atrophy, Huntington's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, cancer, inflammation, Crohn's disease, heart, aging, or traumatic brain injury (TBI).

The present invention also describes the use of at least one compound or composition described in the present invention in the manufacture of a medicament for the treatment of a pathological condition associated with neurodegeneration or protein aggregation/accumulation. In some embodiments, such a condition is a neurodegenerative disease or condition. In some embodiments, said condition is Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Lewy body dementia, Parkinson's disease dementia, multiple system atrophy, Huntington's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, cancer, inflammation, Crohn's disease, heart, aging, or traumatic brain injury (TBI).

The present invention also describes methods of preventing aggregation or accumulation or improving clearance of a protease resistant protein, which comprises contacting the protease resistant protein with an effective amount of at least one compound or composition of the present invention. In some embodiments, contact occurs in vitro or ex vivo. In some embodiments, contact occurs in vivo.

The present invention also describes methods for reducing neuroinflammation in a patient. In some embodiments, the present invention describes a method of reducing neuroinflammation in a patient, comprising administering to the subject an effective amount of a compound or composition of the present invention. In some embodiments, methods for treating a disease or condition associated with neuroinflammation are described, comprising administering to a subject an effective amount of a compound or composition of the present invention. In some embodiments, the use of at least one compound or composition of the present invention in the manufacture of a medicament for reducing neuroinflammation in a patient is described. In other embodiments, the use of at least one compound or composition of the present invention in the manufacture of a medicament for the treatment of a disease or condition associated with neuroinflammation is described.

In one aspect, kits containing a compound or composition of the present invention and instructions for use are described. The kits may contain instructions for use in the treatment of a condition in a patient in need of such treatment. In some embodiments, the pathological condition is a neurodegenerative disease or condition. In some embodiments, said condition is Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Lewy body dementia, Parkinson's disease dementia, multiple system atrophy, Huntington's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, cancer, inflammation, Crohn's disease, heart, aging, or traumatic brain injury (TBI). Said kit may further comprise any materials or devices that can be used to administer the compound or composition, such as vials, syringes, or IV bags. Said kit may also contain sterile packaging.

Chemical synthesis.

Embodiments of the present invention also relate to methods and intermediates that can be used to obtain the compounds of the present invention, as well as their salts or solvates.

Much general literature is known containing well-known chemical syntheses and conditions that can be used to synthesize the disclosed compounds (see, for example, Smith and March, March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Fifth Edition, Wiley-Interscience, 2001 ).

The compounds described in the present invention can be purified by any of the methods known in the art, including chromatographic methods such as high performance liquid chromatography (HPLC), preparative thin layer chromatography, flash column chromatography, and ion exchange chromatography. Any suitable stationary phases may be used, including normal and reversed phases, as well as ion exchange resins. In the most typical case, the compounds described in the present invention are purified by chromatography on silica gel and/or alumina. See, for example, Introduction to Modern Liquid Chromatography, 2nd ed., ed. L.R. Snyder and J.J. Kirkland, John Wiley and Sons, 1979; and Thin Layer Chromatography, E. Stahl (ed.), Springer-Verlag, New York, 1969.

During any of the processes for preparing the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules. This can be done using well-known protecting groups as described in well-known works such as the T/W book. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 4th ed., Wiley, New York, 2006. Protective groups may be removed at a suitable subsequent step using methods known in the art.

Examples of chemical compounds that can be used in the methods of the present invention will be described below using illustrative synthesis schemes in the form of a General description and specific examples of obtaining. Those skilled in the art will appreciate that, in order to prepare the various compounds described herein, the starting materials must be properly selected so that the desired substituents proceed through the reaction scheme with or without protection groups to yield the desired product. Alternatively, it may be necessary or desirable to use a suitable group in place of the final desired substituent, which can be carried through the reaction scheme and then replaced by the desired substituent. In addition, the skilled artisan will appreciate that the transformations shown in the diagrams below may be performed in any order that is compatible with the functionality of each particular group. Each of the reactions depicted in the general schemes is carried out at a temperature from about 0° C. to the boiling point of the organic solvent used. Unless otherwise indicated, all variables have the meanings given above for formula (I). The isotopically labeled compounds described herein are prepared according to the methods described below, using the appropriate labeled parent compounds. Such substances are generally available from commercial suppliers of radioactively labeled chemicals.

Representative syntheses of compounds having formula (I) are described in Schemes 1 and 2.

Stage 4

Fe, aq. HC!

NO ₂ EtOH

Stage 5

Stage 8

CHOEt ₃

Scheme 1

Scheme 2

In schemes 1 and 2 R ¹ , R ² and R ³ have the meanings specified in the present text. Starting compounds can be obtained from commercial sources or by well known synthetic methods.

Scheme 3 shows the general synthesis of compounds according to one of the embodiments of formula (I).

Scheme 3

In Scheme 3, R ¹ , R ² , R ³ , R ^y and R ^z are as defined herein.

In some cases, the above methods further include the step of forming a salt of the compound of the present invention. Some embodiments relate to other methods described in the present invention and the product obtained by any of the methods described in the present invention.

- 29 042432

Examples

The following examples are offered to illustrate and not limit the present invention. Compounds were prepared using the general methods described above.

The following chemical abbreviations are used in the examples:

ACN - acetonitrile, (BPin) ₂ - bis (pinacolato) diborone,

DXM - dichloromethane,

DMF - dimethylformamide,

DMSO - dimethyl sulfoxide,

EDTA - ethylenediaminetetraacetic acid,

EtOH - ethanol,

HPLC - high performance liquid chromatography,

IPA - isopropyl alcohol,

IPAc - isopropyl acetate,

LCMS - liquid chromatography - mass spectrometry, mCPBA - meta-chloroperoxybenzoic acid,

MeOH - methanol,

MTBE - methyl tert-butyl ether,

THF - tetrahydrofuran,

2-MeTHF is 2-methyltetrahydrofuran, and p-TSA or TsOH is p-toluenesulfonic acid.

Example 1 4-(4-((4-Chloro-3-(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3thione (compound 1).

(i) Synthesis route A.

Stage 1

To a mixture of 4-chloro-3-(trifluoromethyl)aniline (500 g, 2.56 mol) in HCl (750 ml) and H2O (750 ml) was added dropwise a solution of NaNO2 (194 g, 2.81 mol) in 250 ml of water, maintaining the temperature below 5°C. The mixture was stirred at 0-5° C. for 30 minutes. A solution of potassium ethoxycarbothioylsulfanyl (492 g, 3.07 mol) in 1 L of water was added dropwise at 0-5°C and the resulting mixture was stirred at 20°C for 12 hours. The mixture was extracted with ethyl acetate (1 L, 3 times). The organic layers were washed with saturated sodium chloride solution (1 L), dried over Na ₂ SO ₄ and evaporated to give o-ethyl [4-chloro-3 (trifluoromethyl)phenyl]-sulfanylmethanethioate (600 g, crude product) as a brown oil, which was used directly in the next step.

KOH (470 g, 8.38 mol) was added to a mixture of o-ethyl [4-chloro-3-(trifluoromethyl)phenyl]sulfanylmethanethioate (600 g, 2 mol) in EtOH (2 L) and H2O (200 ml). The mixture was stirred at 80°C for 12 hours LCMS showed the presence of the target compound. The EtOH was evaporated to give a brown residue which was dissolved in H2O (2 L) and extracted with 1:1 MTBE/petroleum ether (1 L, 3 times). The aqueous layer was adjusted to pH 1 with concentrated HCl and extracted with ethyl acetate (1 L, 2 times). The organic layers were washed with saturated sodium chloride solution (1 L), dried over Na ₂ SO ₄ and evaporated to give 4-chloro-3-(trifluoromethyl)benzenethiol (480 g, crude) as a brown oil.

Stage 2

Cs ₂ CO ₃ (1.15 kg, 3.53 mol) and 1-fluoro-4-nitro-benzene (300 g , 2.12 mol). The mixture was stirred at 80° C. for 3 hours. The mixture was filtered, water was added to 3 L and extracted with ethyl acetate (1 Lx3). The organic layer was washed with 2 L of saturated aqueous sodium chloride, dried over Na ₂ SO ₄ and evaporated to give (4-chloro-3-(trifluoromethyl)phenyl)(4-nitrophenyl)sulfane (640 g, crude) as a brown solid substances.

¹ H NMR (CDCl3, 400 MHz) δ 8.13-8.16 (m, 2H), 7.83 (d, J=0.8 Hz, 1H), 7.58-7.60 (m, 2H), 7.27-7.29 (m, 2H).

- 30 042432

Stage 3a.

mCPBA (822 g, 4.05 mol, 80% purity) was added to a mixture of A-3 (640 g, 1.93 mol) and DCM (3.5 L) at 20°C. The mixture was stirred at 20°С for 12 h. The resulting mixture was added to a solution of Na ₂ SO ₃ (100 g, 0.79 mol) and Na ₂ CO ₃ (250 g, 2.36 mol) in 4 L H2O and stirred at 20°С in for 2 hours. The mixture was filtered and the solid product was separated as the target compound. The aqueous layer was further extracted with dichloromethane (2 Lx2) and the combined organic layers were evaporated to give a brown solid which was purified by suspension in ethyl acetate (2 L) to give A-4 (475 g, 67% yield) as a white solid.

¹ H NMR (DMSO, 400 MHz) δ 8.33-8.42 (m, 6H), 8.03-8.05 (m, 1H).

Stage 4

HCl (15 ml) was added to a mixture of A-4 (450 g, 1.23 mol) in EtOH (1.25 L) and H ₂ O (1.25 L). The mixture was heated to 70°C. Fe (140 g, 2.46 mol) was added and the resulting mixture was stirred at 70° C. for 3 hours. The mixture was filtered and the EtOH was evaporated. The remaining aqueous solution was extracted with dichloromethane (0.5 Lx3) and the organic layers were evaporated to give a crude solid. The resulting solid was dissolved in DCM (1 Lx3) and the solution filtered. The solvent was evaporated to give the title compound. Pooled portions of A-5 (200 g, 48% yield) were obtained as an earthy yellow solid.

1Н NMR (CDCl3, 400 MHz) δ 8.20 (s, 1H), 7.98 (d, J=7.2 Hz, 1H), 7.70 (d, J= 8.8 Hz, 2H), 7.61 (d, J=8.4 Hz, 1H ), 6.68 (d, J=8.4 Hz, 2H), 4.27 (s, 2H).

Stage 5

To a mixture of A-5 (100 g, 298 mmol) and isopropanol (1.20 L) was added 2-bromo-1,3,4-thiadiazole (49.2 g, 298 mmol) and TsOH-H ₂ O (8.50 g, 44.7 mmol) . The mixture was stirred at 80° C. for 4 hours. The mixture was filtered and the filtrate was evaporated to give the crude product. The resulting crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=3/1~0/1, 0-10% 0.5M NH ₃ H ₂ O/MeOH in DCM) to give a yellow solid which was suspended in MeOH (300 ml), MTBE (500 ml) and H2O (500 ml), then dried in vacuo to give Compound 1 (20 g, 8% yield) as a white solid.

1H NMR (500 MHz, DMSOCH) δ 14.07 (s, 1H), 8.77 (s, 1H), 8.42-8.32 (m, 2H), 8.32-8.25 (m, 2H), 8.03 (dd, J=8.8, 2.3 Hz, 3H).

LCMS ES+ (m/z), 420.0 (M+1) ⁺ , a pattern characteristic of the presence of Cl is observed.

(ii) Synthesis route B.

Stage 1

In a 1 L round bottom flask equipped with a mechanical stirrer and a thermometer, 60 ml of concentrated hydrochloric acid, 60 ml of water, and 4-chloro-3-(trifluoromethyl)benzenamine (19.5 g, 0.1 mol) were added. The mixture was heated to promote dissolution and then cooled to below 0° C. in an ice bath. A solution of sodium nitrite (7.6 g, 0.11 mol) in 10 ml of water was added dropwise, maintaining the internal temperature below 5°C, and the resulting mixture was stirred at 5°C for 30 min. Then the resulting mixture was added to a mixture of potassium ethyl xanthate (19.2 g, 0.12 mol) in 30 ml of water over 2 h. After the reaction was completed (about 30 min), the organic phase was separated and the aqueous layer was extracted twice with diethyl ether. The combined organic layers were washed with 30 ml of 10% sodium hydroxide solution, then with several portions of water until the separated aqueous phase showed a neutral pH value. The organic phase was dried over Na ₂ SO ₄ and evaporated and the resulting residue was dissolved in 95% ethanol (100 ml). The solution was heated to boiling to speed up the dissolution. Potassium hydroxide granules (23.5 g, 0.42 mol) were slowly added to the resulting hot solution so that the solution continued to boil slowly until all substances were completely dissolved in water (about 8 h). Then about 80 ml of ethanol was removed by distillation on a steam bath and the residue was dissolved in a minimum amount of water (about 100 ml). The aqueous solution was extracted with diethyl ether (50 mlx3). The pH value of the aqueous layer was adjusted to 1 by adding 6N. sulfuric acid solution. Extraction was carried out with diethyl ether (50 mlx3), the combined organic layers were dried over Na ₂ SO ₄ and evaporated, obtaining a crude product, which was purified by column chromatography (from 0 to 2% ethyl acetate/petroleum ether), obtaining 4-chloro-3-( trifluoromethyl)benzenethiol (16.1 g, 75%) as a yellow solid.

Stage 2

1-Fluoro-4-nitrobenzene (12.8 g, 0.091 mol) and Cs ₂ CO ₃ ( 59.4 g, 0.182 mol) and the reaction mixture was stirred at 80°C while monitoring by thin layer chromatography (1:30 ethyl acetate/petroleum ether). After completion of the reaction, the mixture was cooled to room temperature and diluted with water (500 ml). The aqueous layer was extracted with ethyl acetate (200 mlx3) and the combined organic layers were washed with saturated sodium chloride solution, dried over Na ₂ SO ₄ , filtered and evaporated to give crude 4-chloro-3-(trifluoromethyl)phenyl)(4-nitrophenyl)sulfane (25 g, 82%) as a yellow oil, which was used in the next step without further purification.

Stage 3b.

To a solution of 4-chloro-3-(trifluoromethyl)phenyl)(4-nitrophenyl)sulfane (25 g, 0.075 mol) in acetic acid (100 ml) was added 30% H ₂ O ₂ ) at room temperature. The reaction mixture was stirred at 85°C while monitoring by thin layer chromatography (1:5 ethyl acetate/petroleum ether). After completion of the reaction, the reaction mixture was quenched with water. The aqueous layer was extracted with ethyl acetate (100 mlx3) and the combined organic layers were washed with brine, dried over Na2SO4, filtered and evaporated to give crude product which was purified by flash chromatography (0 to 10% ethyl acetate/petroleum ether) to give 1 -chloro-4-(4-nitrophenylsulfonyl)-2-(trifluoromethyl)benzene (20.8 g, 76%) as a white solid.

B-4 B-5

Stage 4

Five drops of concentrated HCl were added to a mixture of iron powder (16 g, 0.29 mol), water (100 ml), and ethanol (100 ml). The mixture was heated to boiling by adding 1-chloro-4-(4-nitrophenylsulfonyl)2-(trifluoromethyl)benzene (26.4 g, 0.072 mol). The reaction mixture was refluxed for an additional 1 hour while monitoring by thin layer chromatography (1:5 ethyl acetate/petroleum ether). After completion of the reaction, the hot mixture was filtered and the filter cake was washed with ethanol. The pH of the filtrate was adjusted to a value of 10 by adding 2N. NaOH solution and the aqueous phase was extracted with ethyl acetate (100 mlx3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated to give crude product which was purified by flash chromatography (0 to 15% ethyl acetate/petroleum ether) to give 4-((4-chloro -3-(trifluoromethyl)phenyl)sulfonyl)aniline as a white solid (19.4 g, 79%).

- 32 042432

Stage 6

Thiophosgene (6.6 g, 0.057 mol) was added to a two-phase solution of 4-((4-chloro-3(trifluoromethyl)phenyl)sulfonyl)aniline (19.2 g, 0.057 mol) in dichloromethane and water containing sodium bicarbonate (13.4 g, 0.13 mol ), at 0°С. The reaction mixture was stirred at 0°C for 2 hours. After the end of the reaction, the organic layer was separated, dried over Na ₂ SO ₄ , filtered and evaporated to dryness. The residue was purified by column chromatography (0 to 50% ethyl acetate/petroleum ether) to give 1-chloro-4-(4-isothiocyanatophenylsulfonyl)-2-(trifluoromethyl)benzene (11.5 g, 53%) as a yellow solid.

Stage 7

Hydrazine monohydrate (5.2 g, 0.058 mol) was added dropwise to a solution of 1-chloro-4-(4isothiocyanatophenylsulfonyl)-2-(trifluoromethyl)benzene (11 g, 0.029 mol) in ethanol (60 ml) at 0°C. After 4 h the reaction mixture was diluted with water (100 ml) and extracted with dichloromethane (50 mlx3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated to give crude N-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)phenyl) hydrazinecarbothioamide (8.4 g, 70 %), which was used in the next step without further purification.

B-7 Compound 1

Stage 8

N-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)phenyl) hydrazinecarbothioamide (8.2 g, 0.02 mol) and triethoxymethane (50 ml) were heated at 145°C for 3 h. Water (100 ml) and the resulting mixture was extracted with dichloromethane (50 mlx3). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and evaporated to give the crude product, which was purified by column chromatography (0-10% ethyl acetate/petroleum ether) to give the title compound (5.4 g, 64 %) as a white solid.

¹ H NMR (300 MHz, DMSO-d6) δ 14.07 (s, 1H), 8.77 (s, 1H), 8.41-8.32 (m, 2H), 8.32-8.25 (m, 2H), 8.06-8.00 (m, 3H).

LCMS ES+ (m/z), 420.0 (M+1) ⁺ , a pattern characteristic of the presence of C1 is observed.

In FIG. 1B shows the 2D NOESY spectrum of compound 1 in DMSO-d ₆ (400 MHz) synthesized via pathway B. FIG. 1C shows a stretched region of the 2D NOESY spectrum of compound 1 in DMSO-d ₆ (500 MHz) synthesized via route B. The NOESY spectra show the nOe interaction between the triazolethione CH and the phenyl CH corresponding to R ¹ in formula 1.

(iii) Synthetic route C.

s ^F 3C^^ _{NH 2}

WADN. HCl, NaNO^ _

Cl Cl

C' ¹ Stage-1^ ²

Stage 1

A magnetic stirrer was placed in a 250 ml flask with a thermal jacket. The flask was loaded with concentrated HCl (25 ml, 0.30 mol, 3.0 equiv.) and water (98.2 ml). 4-Chloro-3-(trifluoromethyl)aniline (20.0 g, 0.10 mol, 1.0 equiv.) was melted and added to the flask at 25°C. The mixture was heated to 50°C and stirred at 50°C for 30 minutes. After cooling the mixture to 0–5°C, a solution of NaNO ₂ (7.6 g, 0.11 mol, 1.1 equiv.) in 12 ml of water was added dropwise over 30 min, maintaining the temperature in the range of 0–5°C. After the addition of NaNO ₂ was completed, the mixture was stirred at 0-5°C for 1 h.

The second reaction flask was charged with potassium ethyl xanthate (20.8 g, 0.13 mol, 1.3 eq.) and then water (80 ml). After stirring for 20 minutes, toluene (80 ml) was added and then the diazonium salt from the first reaction flask was added dropwise at 19-23°C over 3 hours. After the addition was complete, the mixture was stirred at 20°C for 2 hours. the phase was separated from the organic phase and extracted with 20 ml of toluene three times. The organic phases were combined and washed with water (10 ml, 4 times) and

- 33 042432 was then degassed by passing through a nitrogen solution for 30 minutes.

The third flask was charged with EtOH (63.2 g), water (10 ml), and KOH (23.0 g, 0.41 mol, 4.1 equiv.). The resulting ethanolic KOH solution was degassed by bubbling nitrogen through the mixture for 30 min. The resulting KOH solution was heated to 75-82° C. under an inert nitrogen atmosphere. The toluene solution from the second reaction flask was added to the degassed ethanolic KOH solution at 75-82°C for 2 hours under an inert nitrogen atmosphere. After the addition, the mixture was stirred at 78°C for 3.5 h.

The mixture was distilled off, reducing the volume by a factor of 1.5–2 at 45°С. More toluene (60 ml, degassed with nitrogen bubbling) was added to the resulting mixture, then distilled off again, reducing the volume by a factor of 1.5-2 at 45°C, and toluene (20 ml, degassed with nitrogen bubbling) was added. Water (80 ml, degassed with nitrogen bubbling) was added to the reaction flask and the aqueous phase was separated from toluene. The aqueous phase was washed 3 times with 20 ml of toluene. The aqueous phase was cooled to 10°C and the pH value was brought to pH<1 by adding concentrated HCl (32.0 ml) at 10-15°C. The mixture was purged with nitrogen for 20 min and heated to 20°C. MTBE (40 ml, nitrogen degassed) was added under a nitrogen atmosphere. The organic and aqueous phases were separated. The aqueous phase was extracted with MTBE (40 ml, nitrogen degassed) 3 times. The organic MTBE phases were combined and washed with water (10 ml, nitrogen degassed) 3 times. According to HPLC data, the yield of 4-chloro-3-(trifluoromethyl)benzenethiol was 64.5%. The product was then transferred from MTBE to acetonitrile by distillation at 60° C. at atmospheric pressure. Acetonitrile (50 ml) was added and the resulting mixture was distilled at 80° C. under atmospheric pressure. Additional acetonitrile (40 ml) was added to give 4-chloro-3-(trifluoromethyl)benzenethiol with no residual MTBE.

Stage 2

Cs ₂ CO ₃ (195.0 g, 0.60 mol, 2.1 equiv.) and 1-fluoro-4 -nitro-benzene (52.3 g, 0.37 mol, 1.3 equiv.). The mixture was stirred at 80°C for 11 hours, cooled to 25-30°C and filtered. The filter cake was washed with acetonitrile (120 mlx2). The acetonitrile solution was evaporated to 60-120 ml under reduced pressure, keeping the temperature below 45°C. Dichloromethane (1116 ml) and 15% NaCl solution (1600 ml) were added to the solution. The mixture was stirred at 20-30°C for 30 min and the organic layer was separated. The organic layer was washed with 5% NaCl solution 2 more times. The organic layer was evaporated to 480-600 ml under reduced pressure, keeping the temperature below 45°C. Dichloromethane (560 ml) was added to the solution and the organic layer was evaporated again to 480-600 ml to give a solution of (4-chloro-3(trifluoromethyl)phenyl)(4-nitrophenyl)sulfane in DCM, which was used directly in the next step.

Stage 3

An additional amount of DCM (340 ml, 20 vol.) was added to dichloromethane (8.5 vol.) stage 2. The mixture was heated to 33-37°C and stirred for 0.5 h, then m-CPBA (31.0 g, 152.8 mmol, 3.0 eq., 85 wt.%) was added in portions at 33-37°C. The mixture was stirred at 33-37°C for 4 hours and then cooled to 20-30°C. A 16% aqueous solution of Na ₂ SO ₃ (146.2 g, 8.6 X) and a 16% aqueous solution of Na ₂ CO ₃ (146.2 g, 8.6 X) were added to the resulting mixture, maintaining the temperature below 30°C. The mixture was stirred at 20-30°C for 1 h. The organic layer was separated, washed with 10 wt.% NaCl solution (51.0 g, 3 X) and evaporated to 3-5 vol. under reduced pressure at temperatures below 45°C. IPAc (15 vol.) was added and the solution was evaporated to 6-8 vol. under reduced pressure at temperatures below 45°C. IPAc (15 vol.) was added to the mixture a second time, then the solution was again evaporated to 6-8 vol. under reduced pressure at temperatures below 45°C. Added IPAc (28 vol.) and heated the resulting mixture at 60°C with stirring, obtaining a clear solution. The solution was cooled to 55°C with stirring for 1-2 hours. The solution was distilled off to 3-5 vol. under reduced pressure at temperatures below 55°C. The mixture was cooled to 45°C for 2 hours. MTBE (11 vol.) was added to the mixture and the resulting mixture was stirred at 45°C for another 1-2 hours. The mixture was cooled to -10°C for 11 hours and kept at -10°C for another 4.5 hours. The mixture was filtered and the filter cake was washed twice with a mixture of 1PAc/MTBE=1/4 (4 vol.). The wet filter cake was dried for 1 hour under reduced pressure at a temperature below 45°C, obtaining

1-Chloro-4-((4-nitrophenyl)sulfonyl)-2-(trifluoromethyl)benzene (19.5 g, 99.7% yield) as an off-white solid (97.5% pure).

Stage 4

1-Chloro-4-((4-nitrophenyl)sulfonyl)-2-(trifluoromethyl)benzene (20.0 g, 54.7 mmol) and IPAc (200 ml) were added to a pressure reaction vessel having a volume of 1 L. The vessel was purged and degassed with argon, placed in it with 5% Pt/C (800 mg) under nitrogen atmosphere, purged and degassed with hydrogen, and the resulting mixture was stirred at 0.5 MPa (72.5 psi) H ₂ pressure at 65°C in during 18 h. During this time, the hydrogen pressure dropped to 0 MPa, so H2 was pumped into the vessel to a pressure of 0.5 MPa and kept at 65°C for 14 h. , yielding a light yellow solid (18.0 g, 98.5% crude yield).

HCI/NaNO ₂ _____ S^ _N

H ₂ N ^N CuCI (cat.) CI^ ^N

C-6 07

Stage 5

To a flask containing 1,3,4-thiadiazol-2-amine (5.0 g, 49.4 mmol) at 30°C was added 30 ml of HCl (30 g, 36.5% aq., 300 mmol), then 25 ml of H2O. The solution was cooled to 0°C, obtaining a suspension. CuCl (0.5 g, 4.9 mmol) was added at 0°С. A solution of NaNO ₂ (3.4 g, 49.4 mmol) in H2O (50 mL) was added slowly at 0°С over 30 min, and the reaction mixture was stirred for 2.5 h at 0–5°С. IPAc (100 ml) was added and the reaction was quenched with 10% NaHSO ₃ solution (60 ml). NaHCO ₃ (25 g, solid) was added slowly until pH 6-7 and the organic layer separated. The aqueous layer was extracted with IPAc (100 mlx2). The organic layers were combined and washed with 10% EDTA (50 mlx4) and H2O (100 ml). The combined EDTA aqueous layers and H ₂ O layers were extracted with IPAc (100 ml). The combined organic IPAc extracts were dried over Na ₂ SO ₄ , filtered, evaporated in vacuo, redissolved in IPAc (100 ml) and evaporated in vacuo (2x) to give the crude product (4.0 g) as a light yellow oil. The resulting oil was stored at 5°C for up to 12 hours.

Stage 6

4-((4-Chloro-3-(trifluoromethyl)phenyl)sulfonyl)aniline (7.0 g, 20.9 mmol) and IPA (93 ml) were added to the reaction vessel at 30° C. to give a suspension. Added p-TSA.H ₂ O (595 mg) and heated the reaction mixture to 80-85°C. 2-Chloro-1,3,4-thiadiazole (4.6 g, 38.2 mmol) in IPA (20 ml) was added at 80-85°C over 5 h and the resulting mixture was stirred for 1 h after the addition was complete. The mixture was cooled to 30° C. and left for 15 hours. The reaction mixture was evaporated to dryness. MTBE (50 ml) was added, the resulting mixture was stirred for 2 h at 30° C. and filtered. The MTBE layer contained 1.0 g (3%, 32 gx3%=1.0 g) 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2 ,4-triazole-3thione. The filter cake was taken up in 100 ml 2-MeTHF and saturated NaHCO ₃ solution was added to pH 7-8. The yield in the 2-MeTHF layer included 4.1 g (102 gx4%=4.1 g) 4-(4-((4-chloro-3(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2 ,4-triazole-3-thione. The total weight was=5.1 g, 58% yield.

^1H NMR (400 MHz, DMSOL) δ 14.09 (s, 1H), 8.78 (s, 1H), 8.38-8.35 (m, 2H), 8.30-8.28 (m, 2H), 8.03 (m, 3H).

LCMS ES+ (m/z), 420.0 (M+1) ⁺ , a pattern characteristic of the presence of Cl is observed.

The 1H NMR spectrum of compound 1 is shown in FIG. 1A.

In FIG. 1D shows the 2D NOESY spectrum of compound 1 in DMSO-66 (400 MHz) synthesized by Route C. The NOESY spectra show the nOe interaction between the triazolethione CH and the phenyl CH corresponding to R ¹ in formula 1. FIG. 1E shows the HMWR spectrum of compound 1 in DMSO-66 (400 MHz) showing the correlation between the CH of the triazolethione and the aromatic carbon attached to the triazolethione.

- 35 042432

(iv) Synthesis route D.

Stage-1 ^{i) BoflHHci} ' ^NaNo 2 ^:v 2) A to

D-1 EtO^S ^K

D-2

3) KOH, EUN

4) HCI, MTBE

Stage 1

Purified water (178 kg) was placed in a reaction vessel, then concentrated HCl (216 kg) and 4-chloro-3-(trifluoromethyl)aniline (60.55 kg, 1.0 eq.) were added. The mixture was heated to 45-55°C, stirred for 5 hours and then cooled to -5~5°C. A solution of NaNO2 (25.65 kg) was added dropwise to 38 kg of water for 1–2 h at –5~5°C. After the addition, the mixture was stirred at 0–5°C for 2 h. for 2-6 h in a reactor containing toluene (211.6 kg, 4 vol.) and 0.5 volume of purified water. The resulting mixture was stirred at 20° C. for 5-12 hours. The layers were separated and the aqueous phase was extracted with toluene (112 kg). The organic layers were combined and washed with purified water 3 times.

Ethanol (208 kg) and water (32 kg) were charged to the second reaction vessel, then KOH (71 kg) was added. The mixture was heated to 75-82° C. under nitrogen. The toluene solution from the extraction was added at 75-82° C. under nitrogen for 5 hours. The mixture was stirred at 78° C. for 5 hours. The mixture was then distilled to 2-4 vol. At an internal temperature not exceeding 45°C and again distilled with toluene (169 kg) to remove EtOH. Purified water (250 kg) was loaded into the vessel with stirring; the toluene phase was separated and the aqueous layer was washed with 2 volumes of toluene to give a product-rich aqueous layer.

The aqueous layer was cooled to 0-10° C. and nitrogen was bubbled through it for 2 h, during which time 6 N purged with nitrogen was added dropwise. HCl (2.0-5.0X) at 0-10°C until the pH was from 1 to 2. The mixture was stirred for 1 hour at 0-10°C. The resulting mixture was stirred for 1 h at 0-10°C and then extracted with MTBE (250 kg) which was also purged with nitrogen for 2 h. The organic layer was separated and washed with purified water twice (2x268 kg) and the resulting organic layer kept for further work with him. 36.6 kg of 4-chloro-3-(trifluoromethyl) benzenethiol (D-2) was obtained as a solution in MTBE. The product was a mixture of monomer and dimer in 55.5% yield.

Stage-2 fY /bNO ₂ F ₃ C_^_S_^ ¥ ¥ . ¥ ¥ ¥ Ί

Acetonitrile

D-2 Cs ₂ CO ₃ D-3

Stage 2

A mixture of D-2 and dimer (34.1 kg, 158.8 kgx21.5 wt.%, 1.0 eq.) in MTBE (3 vol.) Was loaded into the reaction vessel. Acetonitrile (482 kg, 18.6 vol.) was added, followed by Cs ₂ CO ₃ (157 kg, 3.0 eq.) and 1-fluoro-4-nitrobenzene (29.6 kg, 1.3 eq.). The mixture was heated to 60-65°C and stirred at this temperature for 57 hours. The mixture was cooled to 20-30°C. Celite (37 kg) was added and after stirring for 1-3 h the mixture was filtered and washed with acetonitrile (163 kg). The resulting acetonitrile solution was evaporated to 6-7 volumes at a temperature below 45°C in vacuum. Then the mixture was stirred at 40-45°C for 0.5-1 h until a clear solution was obtained. The mixture was cooled to 25-30°C for 1-2 h and then stirred for another 0.5-1 h. Seed crystals D-3 (96 g) were added and the resulting mixture was stirred for 1-2 h. Water (136 kg) was added dropwise for 7 h and the resulting mixture was stirred at 25-30°C for 10-20 h. The mixture was centrifuged and the resulting pellet was washed twice with ACN/H2O (104 kg, 6:4 v/v). The wet precipitate was dried at 50-60°C for 24 h, obtaining 40.4 kg of (4-chloro-3(trifluoromethyl)phenyl)(4-nitrophenyl)sulfane (D-3) with a yield of 74.4%.

Stage 3

DCM (1480 kg) was charged into the reaction vessel, then 40.4 kg of D-3 was added. The mixture was heated to 33-37°C. MCPBA (3x20.6 kg) was added in portions at 33-37°C and stirred for 20-30 min between additions. After the addition was completed, the reaction mixture was stirred for 3-5 hours at 33-37°C. After cooling to 20-30°C, 16% aqueous solution of Na ₂ SO ₃ (344 kg) and 16% aqueous

- 36 042432 Na ₂ CO ₃ solution (342 kg). The mixture was stirred for 1-2 hours and then extracted with dichloromethane (342 kg). The organic layer was separated and washed with 7% Na ₂ SO ₄ aqueous solution (134 kg) 2 times. The organic layer was evaporated to 3-4 vol. under reduced pressure at a temperature below 35°C, washing the substances from the walls of the reaction flask with dichloromethane (114 kg). MTBE (322 kg) was added and the resulting mixture was stirred at 40-50°C for 1-2 h, cooled to 5-10°C and stirred at 5-10°C for 4-6 h. solvents (DCM:MTBE=1:3.118 kg) and suspended in MTBE (156 kg) and DCM (66 kg). After stirring at 5-10° C. for 1-2 hours, the precipitate was filtered off and washed with solvent (DCM:MTBE=1:3, 38 kg). The filter cake was dried under vacuum at

40-45°C for 8-12 h, obtaining 39.87 kg (91.4% yield) of 1-chloro-4-((4-nitrophenyl)sulfonyl)-2(trifluoromethyl)benzene (D-4).

Stage 4

Pt/V/C (2.9 kg) was added to a reaction vessel containing D-4 (38.4 kg) in THF (198 kg) and MeOH (126 kg). The reaction vessel was evacuated and filled with nitrogen 3 times, then evacuated and filled with hydrogen 3 times. The temperature was brought to 60°C and the reaction mixture was stirred in a hydrogen atmosphere (0.3-0.4 MPa) for 17 hours. The reaction mixture was filtered and washed with tetrahydrofuran (97 kg). The filtrate was evaporated to 2-3 volumes. The solvent was replaced by the addition of methanol (120 kg) and evaporation to 2-3 volumes (repeated 3 times). Methanol (64 kg) was added to the reaction vessel and the temperature was brought to 60° C. with stirring for 0.5-1 h. The temperature was lowered to 55° C. and seed crystals D-5 (0.04 kg) were added. The mixture was stirred at 50-60° C. for 5 hours and then lowered to 20° C. over 6 hours. Water (100 kg) was added over 5 hours and then the suspension was stirred for 7 hours. The precipitate was filtered and washed with a solution of MeOH:H ₂ O (3:1, 98 kg). The filter cake was dried in vacuum at 45°C for 16 h to give 4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)aniline, D-5, (31.9 kg) in 90.6% yield.

Stage 5

To a reaction vessel containing NaHCO ₃ solution (23.4 kg) and water (293 kg) was added D-5 (28.5 kg) followed by 361 kg of DCM. After stirring at 15-25°C for 0.5 h, the vessel was cooled to -5~5°C. Twice repeated sequential addition of thiophosgene (12.3 kg, 6 kg) dropwise with stirring at -5~5°C for 4 h and subsequent addition of NaHCO ₃ (3.7 kg, 2.9). A final portion of thiophosgene (6.0 kg) was added, and the reaction mixture was stirred at -5~5°C for 2-10 h, heated to 15-25°C, and stirred for another 1-2 h. The organic layer was separated and washed with water (112 kg). The organic layer was evaporated to 2-3 volumes in vacuum at a temperature below 25°C. The addition of DCM (185 kg) and evaporation (up to 2-3 volumes under vacuum below 25° C.) was repeated 3 times with a final concentration of DCM of 4-5 volumes. The solvent was replaced by adding portions of dichloromethane solution D-6 to the second reaction vessel, in which 180 kg of methylcyclohexane were placed, with stirring at 20-25°C for 2-4 h and evaporation to 7.5-8.5 volumes in vacuum at a temperature below 25 °C between additions. Methylcyclohexane (2x100 kg) was added to the reaction vessel and the resulting mixture was evaporated to 4.0-4.5 volumes in vacuum at a temperature below 35°C, this procedure was repeated twice. More methylcyclohexane (135 kg) was added and the resulting mixture was stirred at 55~65°C for 3–4 h, slowly cooled (10–12 h) to 0–5°C, and stirred for 6–10 h. The suspension was filtered, washed with 68 kg of methylcyclohexane and dried at 40-50°C for 24 h, obtaining 29.9 kg of 1-chloro-4-((4-isothiocyanatophenyl)sulfonyl)-2-(trifluoromethyl)benzene (D-6) with a yield of 93.2%.

- 37 042432

Stage 6

In a reaction vessel with D-6 (30.95 kg) and DABCO (11.4 kg) was added THF (268 kg) under nitrogen atmosphere. The reaction vessel was cooled to 10~20°C and stirred for 30-60 min, after which formohydrazide (5.6 kg) was added under nitrogen atmosphere. The reaction mixture was stirred at 10~20°C for 1.5 h, heated to 35~45°C, then stirred for 17 h, heated to 45~55°C, and stirred for 9 h. The reaction mixture was cooled to 20~40°C and transferred into the second reaction vessel through a fine filter. The mixture was evaporated to ~2 volumes, keeping the temperature below 40°C. Water (251 kg) was added under nitrogen, then 6N. HCl solution (30.9 kg) to pH 4. The reaction mixture was heated to 40~50°С, stirred for 3 h, then cooled to 15~25°С and stirred for 4 h. The mixture was centrifuged, the precipitate was washed with a mixture of water:THF (3: 1.72 kg) and water (94 kg). The resulting solid product was dried at 40~50°C for 27 hours to give compound 1 in 93.7% yield and 97% purity.

4-(4-((4-Chloro-3-(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione was further purified by filtration and recrystallization. 17.4 kg of 4-(4-((4-chloro-3(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione was dissolved in acetone (158 kg) and stirred at 20-30°C until a clear solution was obtained. The solution was filtered through a fine filter and evaporated 7-9 volumes in vacuo, keeping the temperature below 40°C. The mixture was cooled to 30°C, seed crystals (21 g) were added, stirred for 7 h, then evaporated to 3-5 volumes in vacuo, keeping the temperature below 40°C.

The solvent was replaced twice with ethanol by sequential addition of ethanol (56 kg, 52 kg), stirring and evaporation to 3-5 volumes in vacuum at a temperature below 40°C. The resulting compound was recrystallized from ethanol (88 kg) by heating to 75~82°C, stirring the resulting mixture for 10 h, cooling to 15-25°C for 5 h, and stirring the resulting mixture at 15-25°C for 8 h. The mixture was filtered, washed with 160 grams of ethanol and dried at 40-50°C for 10-16 h, obtaining 16.64 kg of compound 1 with a purity of 99%.

Example 2 4-(4-((4-Chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-morpholinophenyl)-2,4-dihydro-3H1,2,4-triazole-3-thione (compound 2) .

The synthesis of 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-morpholinophenyl)-2,4-dihydro-3H1,2,4-triazole-3-thione was carried out similarly to that described in the synthesis route In example 1 from 4-(5-fluoro-2nitrophenyl)morpholine.

Ή NMR (300 MHz, DMSOCH) δ 14.00 (1H, s), 8.41 (2H, m), 8.67 (1H, s), 8.04 (1H, d, J=6 Hz), 7.90 (1H, dd, J= 3, 6 Hz), 7.83 (1H, d, J=3 Hz), 7.71 (1H, d, J=6 Hz), 3.55 (4H, m), 2.82 (4H, m).

LCMS ES+ (m/z), 505.0 (M+1) ⁺ , a pattern characteristic of the presence of Cl is observed.

Example 3 4-(4-(phenylsulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (Compound 3)

Stage 1. Synthesis of sodium benzenesulfinate.

Phenylsulfonyl chloride (3.5 g, 19.9 mmol, 1 eq.) was added to a solution of sodium sulfite (5 g, 39.8 mmol, 2 eq.) and sodium bicarbonate (3.3 g, 39.8 mmol, 2 eq.) in water (50 ml). The reaction mixture was stirred for 2 hours at room temperature. The water was removed in vacuo, the residue was suspended in methanol and filtered. The residue was washed with methanol 3 more times and filtered. The methanolic filtrates were combined and evaporated. The resulting solid was suspended in methanol and filtered. The filtrate was evaporated to give crude sodium benzenesulfinate which was used in the next step without further purification.

Negative LC-MS: 141.14 (M-H) ^- , C ₆ H ₅ NaO ₂ S.

Stage 2. Synthesis of 4,4,5,5-tetramethyl-2-(4-nitrophenyl)-1,3,2-dioxaborolane.

A mixture of 1-bromo-4-nitrobenzene (2.02 g, 0.01 mol, 1 equiv.), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1 ,3,2dioxaborolane) (2.54 g, 0.01 mol, 1 eq.), potassium acetate (2.88 g, 0.03 mol, 1 eq.) and PdCl ₂ (dppf) (0.82 g,

- 38 042432

1.0 mmol, 0.1 equiv.) in dioxane (35 ml) was boiled overnight. The mixture was cooled to room temperature, diluted with water (100 ml) and extracted with ethyl acetate (100 mlx3). The organic extracts were combined, washed with saturated sodium chloride solution (50 ml), dried over anhydrous sodium sulfate and evaporated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=20:1 to 5:1) to give the product (1.83 g, 73% yield).

Stage 3. Synthesis of 1-nitro-4-(phenylsulfonyl)benzene.

Potassium carbonate (2.01 g, 14.6 mmol, 2 equiv.), 4L molecular sieves and Cu(OAc)2 (1.49 g, 8.0 mmol, 1.1 equiv.) were added sequentially to a solution of 4,4,5,5-tetramethyl-2- (4-nitrophenyl)-1,3,2-dioxaborolane (1.82 g, 7.3 mmol, 1 eq.) and crude sodium benzenesulfinate (2.39 g, 14.6 mmol, 2 eq.) in DMSO (50 ml). The reaction mixture was stirred overnight at 45° C. under oxygen (bead). The reaction mixture was poured into water and extracted with ethyl acetate. The organic extracts were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated. The residue was purified by silica gel column chromatography to give 1-nitro-4(phenylsulfonyl)benzene, 0.71 g, 37% yield.

Stage 4. Synthesis of 4-(phenylsulfonyl)aniline.

1-nitro-4-(phenylsulfonyl)benzene (0.7 g, 2.66 mmol, 1 eq.) was dissolved in acetic acid (10 ml) and Fe (1.49 g, 26.6 mmol, 10 eq.) was added. The reaction mixture was heated at 60° C. for 2 hours. The mixture was cooled to room temperature, diluted with ethyl acetate, filtered and the filter cake was washed with ethyl acetate. The filtrate was washed with saturated sodium chloride solution. The organic extract was evaporated and the residue was purified by silica gel column chromatography to give 4-(phenylsulfonyl)aniline (0.52 g, 2.23 mmol, 84% yield).

Pos. LC-MS: 233.92 (M+H)+, C12H11NO2S.

Stage 5. Synthesis of 1-isothiocyanato-4-(phenylsulfonyl)benzene.

Thiophosgene (308 mg, 2.68 mmol, 1.2 eq.) was added to a mixture of 4-(phenylsulfonyl)aniline (520 mg, 2.23 mmol, 1 eq.) and saturated aqueous sodium bicarbonate (10 ml) in chloroform (10 ml). The reaction mixture was stirred for 2 hours at room temperature under nitrogen. The mixture was extracted with dichloromethane twice. The organic extracts were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated to give 1-isothiocyanato-4-(phenylsulfonyl)benzene, which was used in the next step without further purification.

Step 6. Synthesis of 4-(4-(phenylsulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione.

A solution of crude 1-isothiocyanato-4-(phenylsulfonyl)benzene (275 mg, 1.0 mmol, 1 equiv.) and formohydrazide (60 mg, 1.0 mmol, 1 equiv.) in ethanol (5 ml) was refluxed for 30 min.

The solvent was removed and the residue was dissolved in 2% NaOH solution (5 ml). The reaction mixture was heated at 100° C. for another 2 hours. The mixture was cooled to room temperature and acidified to pH 3-4 with the addition of HCl. The precipitate formed was extracted with dichloromethane twice. The organic extracts were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated. The residue was recrystallized from ethanol to give 4-(4-(phenylsulfonyl)phenyl)2,4-dihydro-3H-1,2,4-triazole-3-thione (48 mg, 0.15 mmol, 15% yield) as not quite white solid.

Negative LC-MS: 316.1 (M-H) ^- , C14H11N3O2S2.

1H NMR (DMSO-de, 400 MHz) δ 14.07 (breadth, 1H), 8.77 (s, 1H), 8.16 (d, J=8.4 Hz, 2H), 7.96-8.15 (m, 4H), 7.64-7.75 ( m, 3H).

Example 4 4-(4-((4-chlorophenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (compound 4).

4-(4-((4-Chlorophenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione was synthesized by a procedure similar to that described for 4-(4-(phenylsulfonyl )phenyl)-2,4-dihydro-3H-1,2,4-triazole-3thione. Yield at step 6: 22%, off-white solid.

Negative LC-MS: 350.0 (M-H) ^- , C14H10CLN3O2S2.

1H NMR (DMSO-d6, 400 MHz) δ 14.08 (broad, 1H), 8.77 (s, 1H), 8.18 (d, J=8.4 Hz, 2H), 7.98-8.06 (m, 4H), 8.79 (d, J=8.4 Hz, 2H).

Example 5 4-(4-((4-Chloro-3-methylphenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (Compound 5)

4-(4-((4-Chloro-3-methylphenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione was synthesized by a procedure similar to that described for 4-( 4-(phenylsulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione. Yield at stage 6: 12%, light yellow solid.

LC-MS: 364.0 (M-H) ^- , C ₁₅ H ₁₂ ClN ₃ O ₂ S ₂ .

1H NMR (DMSO-d6, 400 MHz) δ 14.09 (broad, 1H), 8.77 (s, 1H), 8.17 (d, J=8.4 Hz, 2H), 8.07 (s, 1H), 7.99 (d, J= 8.4 Hz, 2H), 7.86 (d, J=7.6 Hz, 1H), 7.71 (d, J=8.4 Hz, 1H), 2.42 (s, 3H).

Example 6 4-(4-((3-(Trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (Compound 11)

4-(4-((3-(Trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione was synthesized by a procedure similar to that described for 4-(4 -(phenylsulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione. Yield at stage 6: 13%, off-white solid.

Negative LC-MS: 384.1 (M-H) ^- ; C15H10F3N3O2S2.

1H NMR (DMSO-d ₆ , 400 MHz) δ 14.08 (broad, 1H), 8.78 (s, 1H), 8.37 (m, 2H), 8.28 (d, J=8.4 Hz, 2H), 8.14 (d, J =7.6 Hz, 1H), 8.02 (d, J=8.4 Hz, 2H), 7.93 (m, 1H).

Example 7 4-(4-((3-methoxyphenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione (Compound 12)

4-(4-((3-Methoxyphenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione was synthesized by a procedure similar to that described for 4-(4-(phenylsulfonyl )phenyl)-2,4-dihydro-3H-1,2,4-triazole-3thione. Yield at stage 6: 57%, off-white solid.

LC-MS: 346.0 (M-H) ^- , C15H13N3O3S2.

1H NMR (DMSO-06, 400 MHz) δ 14.09 (broad, 1H), 8.78 (s, 1H), 8.19 (d, J=8.4 Hz, 2H), 7.98 (d, J=8.4 Hz, 2H), 7.56 (m, 2H), 7.52 (s, 1H), 7.28 (m, 1H), 3.85 (s, 3H).

Example 8 4-(4-(3-(dimethylamino)phenylsulfonyl)phenyl)-1H-1,2,4-triazole-5(4H)-thione (Compound 15)

Step 1 N,N-dimethyl-3-(4-nitrophenylthio)aniline

- 40 042432

3-Aminobenzenethiol (2 g, 16.0 mmol, 1 equiv.) was added to a mixture of 4-bromonitrobenzene (3.5 g, 16.0 mmol, 1 equiv.) and potassium carbonate (4.4 g, 32.0 mmol, 2 equiv.) in DMF (30 ml ). The reaction mixture was stirred for 2 hours at room temperature. The mixture was poured into water and extracted with ethyl acetate three times. The organic extracts were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=50:1 to 10:1) to give 3-(4nitrophenylthio)aniline (2.74 g, 70% yield).

Pos. LC-MS: 246.7 (M+H)+, C ₁₂ H ₁₀ N ₂ O ₂ S.

¹ H NMR (DMSO-d ₆ , 400 MHz) δ 8.12 (d, J=8.4 Hz, 2H), 7.31 (d, J=8.4 Hz, 2H), 7.16 (m, 1H), 6.75 (s, 1H) , 6.68 (m, 2H), 5.45 (broad, 2H). 3-(4-nitrophenylthio)aniline (1 g, 4.1 mmol, 1 equiv.) was dissolved in acetonitrile (20 ml).

Acetic acid (1 ml) and an aqueous solution of formaldehyde (2.5 ml, 32.0 mmol, 8 equiv.) were added. The solution was stirred for 10 min and NaBH ₃ CN (1.42 g, 20.0 mmol, 5 equiv.) was added. The reaction mixture was stirred for another 2 hours. The mixture was diluted with water and extracted with ethyl acetate three times. The organic extracts were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=200:1 to 100:1) to give N,N-dimethyl-3-(4-nitrophenylthio)aniline (340 mg, 31% yield).

Pos. LC-MS: 274.7 (M+H) ⁺ , C ₁₄ H ₁₄ N ₂ O ₂ S.

Step 2 N,N-dimethyl-3-(4-nitrophenylsulfonyl)aniline

A mixture of N,N-dimethyl-3-(4-nitrophenylthio)aniline (340 mg, 1.24 mmol, 1 eq.) and mCPBA (917 mg, 3.72 mmol, 3 eq.) in dichloromethane (15 mL) was stirred overnight at room temperature. 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.3 g, 4.96 mmol, 4 eq.) was added and the reaction mixture was stirred for another 30 min. The mixture was poured into saturated aqueous sodium bicarbonate and extracted with dichloromethane. The organic extracts were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated to give crude N,N-dimethyl-3-(4-nitrophenylsulfonyl)aniline (400 mg, quant.) which was used in the next step without further purification. .

Step 3 3-(4-aminophenylsulfonyl)-N,N-dimethylaniline.

N,N-dimethyl-3-(4-nitrophenylsulfonyl)aniline (400 mg, 1.3 mmol, 1 equiv.) was dissolved in acetic acid (10 ml) and Fe (728 mg, 13.0 mmol, 10 equiv.) was added. The reaction mixture was heated at 60° C. for 2 hours. The mixture was cooled to room temperature, diluted with ethyl acetate, filtered and the filter cake was washed with ethyl acetate. The filtrate was washed with saturated sodium chloride solution. The organic extract was evaporated and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=6:1 to 3:1) to give 3-(4-aminophenylsulfonyl)-N,N-dimethylaniline (240 mg, 67% yield).

Pos. LC-MS: 276.9 (M+H) ⁺ , C ₁₄ H ₁₆ N ₂ O ₂ S.

Step 4 3-(4-isothiocyanatophenylsulfonyl)-N,N-dimethylaniline.

Thiophosgene (105 mg, 0.91 mmol, 1.1 equiv.) was added to a mixture of 3-(4-aminophenylsulfonyl)-N,Ndimethylaniline (230 mg, 0.83 mmol, 1 equiv.) and saturated aqueous sodium bicarbonate (10 ml) in chloroform ( 10 ml). The reaction mixture was stirred for 2 hours at room temperature under nitrogen. The mixture was extracted with dichloromethane twice. The organic extracts were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated to give crude 3-(4-isothiocyanatophenylsulfonyl)-N,N-dimethylaniline (280 mg, quant.) which was used in the next step. without additional cleaning.

Step 5 4-(4-(3-(dimethylamino)phenylsulfonyl)phenyl)-1H-1,2,4-triazole-5(4H)-thione.

A solution of crude 3-(4-isothiocyanatophenylsulfonyl)-N,N-dimethylaniline (280 mg, 0.9 mmol, 1 equiv.) and formohydrazide (54 mg, 0.9 mmol, 1 equiv.) in ethanol (10 mL) was refluxed for 30 min. The solvent was removed and the residue was dissolved in 2% NaOH solution (10 ml). The reaction mixture was heated at 100° C. for another 2 hours. The mixture was cooled to room temperature and acidified to pH 3-4 with the addition of HCl. The precipitate formed was extracted with dichloromethane twice. The organic extracts were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated. The residue was recrystallized from ethanol to give 4-(4-(3-(dimethylamino)phenylsulfonyl)phenyl)-1H-1,2,4triazole-5(4H)-thione (30 mg, 9% yield).

Negative LC-MS: 360.70 (M+H)+, C16H16N4O2S2.

1H NMR (DMSO-d6, 400 MHz) δ 14.08 (broad, 1H), 8.77 (s, 1H), 8.16 (d, J=12 Hz, 2H), 7.95 (d, J=12 Hz, 2H), 7.40 (m, 1H), 7.19 (m, 2H), 6.99 (m, 1H), 2.97 (s, 6H).

Example 9 3-thione (compound 19).

Stage 1. Synthesis of 1-(5-bromo-2-nitrophenyl)piperidine.

A mixture of 2,4-dibromo-1-nitrobenzene (2.81 g, 10.0 mmol), piperidine (0.94 g, 11.0 mmol), and potassium carbonate (2.76 g, 20.0 mmol) in DMF (20 mL) was heated at 80°C for 3 h. The mixture was cooled to room temperature, diluted with water (100 ml) and extracted with ethyl acetate (100 mlx3). The organic extracts were combined, washed with saturated sodium chloride solution (50 mlx2), dried over anhydrous sodium sulfate and evaporated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=300:1 to 200:1) to give 1-(5-bromo-2-nitrophenyl)piperidine (2.2 g, 77% yield) as a yellow solid.

¹ H NMR (CDCl3, 400 MHz) δ 7.66 (d, J=8.8 Hz, 1H), 7.23 (s, 1H), 7.05 (d, J=8.4 Hz, 1H), 3.03 (m, 4H), 1.71 ( m, 4Н), 1.62 (m, 2Н).

Stage 2. Synthesis of 1-(2-nitro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperidine.

A mixture of 1-(5-bromo-2-nitrophenyl)piperidine (2.2 g, 7.8 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'bi(1 ,3,2-dioxaborolane) (1.97 g, 7.8 mmol), potassium acetate (2.23 g, 23.3 mmol) and PdCl ₂ (dppf) (0.63 g, 0.8 mmol) in dioxane (100 ml) were refluxed overnight.

The mixture was cooled to room temperature, diluted with water (200 ml) and extracted with ethyl acetate (200 mlx3). The organic extracts were combined, washed with saturated sodium chloride solution (50 mlx2), dried over anhydrous sodium sulfate and evaporated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=100:1 to 20:1) to give 1-(2-nitro-5-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)phenyl)piperidine (1.1 g, 43% yield).

Pos. LC-MS: 333.22 (M+H)+, C ₁₇ H ₂₅ BN ₂ O ₄ .

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Step 3. Synthesis of 1-(5-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-nitrophenyl)piperidine.

Potassium carbonate (828 mg, 6.0 mmol), 4A molecular sieves (2 g), and Cu(OAc) ₂ (610 mg, 3.3 mmol) were added sequentially to a solution of compound 2 (1 g, 3.0 mmol) and sodium 4-chloro-3 (trifluoromethyl)benzenesulfinate (1.46 g, 6.0 mmol) in DMSO (25 ml). The reaction mixture was stirred overnight at 60°C under oxygen (balloon). The reaction mixture was poured into water (100 ml) and extracted with ethyl acetate (100 mlx3). The organic extracts were combined, washed with saturated sodium chloride solution (50 mlx2), dried over anhydrous sodium sulfate and evaporated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=200:1 to 80:1) to give 1-(5-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2nitrophenyl)piperidine (110 mg, 8% yield).

Stage 4. Synthesis of 4-(4-chloro-3-(trifluoromethyl)phenylsulfonyl)-2-(piperidin-1-yl)aniline.

1-(5-((4-Chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-nitrophenyl)piperidine (110 mg, 0.24 mmol) was dissolved in acetic acid (10 mL) and Fe (137 mg, 2.4 mmol) was added ). The reaction mixture was heated at 60° C. for 2 hours. The mixture was cooled to room temperature, diluted with ethyl acetate (30 ml), filtered and the filter cake was washed with ethyl acetate (10 ml). The filtrate and washing solution were washed with saturated sodium chloride solution (20 ml). The organic extract was evaporated and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=100:1 to 50:1) to give 4-(4-chloro-3-(trifluoromethyl)phenylsulfonyl)-2-(piperidine-1- yl)aniline (100 mg, quantitative yield).

LC-MS: 418.76 (M+H)+, C18H18CIF3N2O2S.

Step 5. Synthesis of 1-(5-(4-chloro-3-(trifluoromethyl)phenylsulfonyl)-2-isothiocyanatophenyl)piperidine.

Thiophosgene (30 mg, 0.26 mmol) was added to a mixture of 4-(4-chloro-3-(trifluoromethyl)phenylsulfonyl)-2(piperidin-1-yl)aniline (100 mg, 0.24 mmol) and a saturated aqueous sodium bicarbonate solution (10 ml) in chloroform (10 ml). The reaction mixture was stirred for 2 hours at room temperature under nitrogen. The mixture was extracted with dichloromethane (10 mlx2). The organic extracts were combined, washed with saturated sodium chloride solution (10 ml), dried over anhydrous sodium sulfate and evaporated to give crude 1-(5-(4-chloro-3-(trifluoromethyl)phenylsulfonyl)-2isothiocyanatophenyl)piperidine (80 mg, 67 % yield), which was used in the next step without further purification.

Stage 6. Synthesis of 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-(piperidin-1-yl)phenyl)-2,4dihydro-3H-1,2,4-triazole -3 -thion.

A solution of 1-(5-(4-chloro-3-(trifluoromethyl)phenylsulfonyl)-2-isothiocyanatophenyl)piperidine (80 mg, 0.17 mmol) and formohydrazide (10 mg, 0.17 mmol) in ethanol (10 mL) was refluxed for 30 min. The solvent was removed and the residue was dissolved in 2% NaOH solution. The reaction mixture was heated at 100° C. for another 2 hours. The mixture was cooled to room temperature and acidified to pH 3-4 with the addition of HCl. The precipitate formed was extracted with dichloromethane twice. The organic extracts were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and evaporated. The residue was recrystallized from ethanol to give the desired 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2(piperidin-1-yl)phenyl)-2,4-dihydro-3H-1,2 ,4-triazole-3-thione (27 mg, 31% yield) as an off-white solid.

- 43 042432

Pos. LC-MS: 502.88 (M+H)+, C20H18CIF3N4O2S2.

1H NMR (DMSO-d6, 400 MHz) δ 14.01 (broad, 1H), 8.64 (s, 1H), 8.42 (m, 2H), 8.04 (d, J=8.0 Hz, 1H),

7.84 (d, J=8.4 Hz, 1H), 7.77 (s, 1H), 7.67 (d, J=8.4 Hz, 1H), 2.77 (m, 4H), 1.44 (m, 6H).

Example 10 4-(4-(4-Chloro-3-(trifluoromethyl)phenylsulfonyl)-2-(diethylamino)phenyl)-1H-1,2,4triazole-5(4H)-thione (compound 21)

Oh, Oh

4-(4-(4-Chloro-3-(trifluoromethyl)phenylsulfonyl)-2-(diethylamino)phenyl)-1H-1,2,4-triazole-5(4H)thione was synthesized by a procedure similar to that described for 4 -(4-((4-chloro-3(trifluoromethyl)phenyl)sulfonyl)-2-(piperidin-1-yl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione . Yield at stage 6: 12%, off-white solid.

LC-MS: 489.0 (M-H) ^- , C ₁₉ H ₁₈ ClF ₃ N ₄ O ₂ S ₂ .

1H NMR (DMSO-d ₆ , 300 MHz) δ 14.01 (broad, 1H), 8.47 (s, 1H), 8.39 (m, 2H), 8.05 (m, 1H), 7.77 (m, 2H), 7.62 (m , 1H), 2.93 (m, 4H), 0.84 (t, J=6.75 Hz, 6H).

In some cases, the methods described above further include the step of forming a salt of the compound of the present invention. Embodiments relate to other methods described herein and a product obtained by any of the methods described in this text.

In some cases, the methods described above further include the step of forming a salt, including a pharmaceutically acceptable salt, of the compounds of the present invention. Salt forms can be prepared using standard salt preparation techniques known in the art. Embodiments relate to other methods described herein and to a product obtained by any of the methods described herein.

Example 11 Dry spray preparations.

Compound 1 formulations were prepared using dry spray techniques. Four spray solutions containing different polymers in a polymer:compound=3:1 ratio were prepared and sprayed in a Buchi B-290 laboratory spray dryer. The spray parameters and the results obtained are summarized in Table 1. 1.

Table 1

No. SSD Polymer : Compound % solids Approximate flow rate (g/min) Average outlet temperature (°C) Total spray time (min) Exit 1 3:1 PVP-VA 64: Conn. 1 15% 8 40 eleven 90.2% 2 3:1Kollydon 30: Conn. 1 15% 8 41 eleven 87.7% 3 3:1 HPMS E5: Conn. 1 10% 8-10 40 14 76.5% 4 3:1 HPMC- AS: Conn. 1 10% 8 39 18 77.4%

A solution of 80:20 DCM:methanol was used as the spray solvent in all solutions. Spray solutions containing PVP-VA 64 and Kollidon 30 had a solids content of 15% w/w, which includes both polymer and compound content. Spray solutions containing HPMC E5 and HPMC-AS had a solids content of 10% w/w. For each spray cycle, a total of 3.1 g of compound 1 was used.

All SDD dispersions were dried overnight at 40°C under 25 mmHg vacuum with a nitrogen purge for 15-20 min before being removed from the oven for storage under nitrogen in the primary container and dried in the second container.

Compounds and SDD were visualized using polarized light microscopy (PLM) and analyzed by powder X-ray diffraction analysis (PXRD), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA).

PXRD was performed using a Rigaku powder X-ray diffractometer (MiniFlex 600 FAE-R PDXL-Version 2-0 Cu Ka radiation S/N BD63000375). In FIG. 2A shows a PXRD (powder X-ray diffraction analysis) diagram of compound 1. The PXRD diagram of compound 1 shows that this compound is mostly crystalline because the peaks are sharp. In FIG. 2B shows an overlapping PXRD plot of four different dry spray preparations of Compound 1. The PXRD charts of the dry spray dispersions (SDD #1-4) show that the dry spray dispersions are predominantly amorphous.

In FIG. 3A shows the overlap of DSC and TGA thermograms of compound 1. FIG. 3B, 3D, 3F and 3H

- 44 042432 shows TGA thermograms of dispersions obtained by the method of dry spraying (SDD) #1-4, respectively. In FIG. 3C, 3E, 3G, and 3I show DSC thermograms of SDD #1-4 dispersions, respectively.

Pharmacokinetic (PK) characteristics of three different Compound 1 formulations (free base and two dry spray dispersions, SDD #1 and SDD #3) were evaluated in male Sprague Dawley (CD®IGS) rats after a single gavage (PO, orally) 30, 100 or 500 mg/kg in a volume of 10 ml/kg. A total of 45 animals were tested in this study (5 rats at doses of 3 dose levels of 3 of the drug). The vehicle consisted of 0.75% hydroxypropyl methylcellulose (HPMC; w/v), 0.2% Tween 20 (v/v) and deionized water. In FIG. 4A shows the PK curves of Compound 1 in free base (FB) form and two dry spray dispersions of Compound 1 (SDD #1 and SDD #3). In FIG. 4B shows area under the curve (AUC) versus Compound 1 free base (FB) dosage and two Compound 1 dry spray dispersions (SDD #1 and SDD #3).

Example 12. X-ray diffraction analysis of single crystals.

Single crystal X-ray diffraction analysis (SXRD) was performed (Solid Form Solutions, Penicuik, Scotland, UK) to determine the structure of compound 1, and the results are summarized in table. 2 and 3. X-ray diffraction analysis of single crystals was carried out on an Agilent SuperNova instrument with a dual source, at 120 K using Mo Ka radiation (λ=0.71073 A) generated by a sealed tube. The obtained data were corrected for absorption effects using an empirical correction for spherical harmonics. All data were processed, resolved, and refined in the achiral triclinic space group P-1.

Compound 1 (about 10 mg) was dissolved in isopropyl acetate (500 μl) in a 2 ml clear glass HPLC vial, and then heptane diffused slowly into the solution of compound 1 at room temperature. After standing at room temperature for several days, growth of large crystals in the form of blocks below the meniscus of the solution was observed, which proved to be suitable for examination by single crystal X-ray diffraction analysis.

A colorless fragment of the plate (0.237x0.158x0.126 mm) was used for X-ray diffraction analysis of the single crystal. The crystal was coated with Paratone oil and examined on a Rigaku Oxford Diffraction (Dual Source) SuperNova diffractometer with Mo Ka radiation (λ=0.71073 A, 40 kV/40 mA) with a graphic monochromator at 120(1) K, with an Oxford Cryosystems 700+ low-temperature apparatus and detector Atlas CCD (Rigaku Oxford Diffraction). A total of 2123 reflections were recorded for the reflection hemisphere, with ω strategy calculated on CrysAlisPro (Rigaku Oxford Diffraction 1.171.38.43h, 2015) in the range of θ 3.02-31.25° with a step size of 1° and exposure of 20 s per reflection. Reflections were integrated with CrysAlisPro (Rigaku Oxford Diffraction 1.171.38.43h, 2015) into a triclinic unit cell using a moving averaged background, which gave a total of 106625 reflections, of which 10259 were independent (I>2σ(I)). The obtained data were integrated up to 2θmax=62.5° (95.4% completeness). Absorption was corrected using SCALE3 ABSPACK (CrysAlisPro 1.171.38.43h, Rigaku Oxford Diffraction, 2015) using an empirical spherical harmonic model combined with Gaussian integration over a multidimensional crystal model (absorption coefficient G=0.533 mm-1).

The OLEX2 software package (Dolomanov, O.W., Bourhis, L.J., Gildea, R.J., Howard, J.A.K., Puschmann, H.J Appl. Cryst., 2009, 42, 339-341) was used as an interface for phase determination and structure refinement. Results were processed with Superflip (Palatinus, L. & Chapuis, G. (2007), J. Appl. Cryst., 40, 786-790; Palatinus, L. & van der Lee, A. (2008), J. Appl Cryst., 41, 975-984; Palatinus, L., Prathapa, S. J. & van Smaalen, S. (2012), J. Appl. Cryst., 45, 575-580) and F2 least squares (Sheldrick, G.M. (2015), Acta Cryst C71, 3-8) in the triclinic space group P-1. A higher level of symmetry was searched using the ADDSYMM function (Le Page, Y.J. Appl. Cryst. 1987, 20, 264; Le Page, Y.J., Appl. Cryst., 1988, 21, 983) in PLATON (Spek, A.L., Acta Cryst., 2009, D65, 148), but a higher level of symmetry has not been found. All non-hydrogen atoms were localized on a Fourier map and their positions were refined anisotropically before calculating their temperature fluctuations for all non-hydrogen atoms. Within the asymmetric cell, two complete crystallographically independent formula units of compound 1 were found, one of which (the 'B' molecule) showed disordering about three positions. The disorder was refined with the SHELX compatible SUMP command and the three fragments showed an occupancy of 34.1:43.2 22.7%. In addition, disordered C11B rings (C12B, C13B, C14B, C9B, C10B); C11C (C15D, C13C, C14C, C9C, C10C); C11D (C12D, C13D, C14D, C9D, C10D) were refined as rigid hexagons using the AFIX66 SHELX compatible command. In addition, C15B-C13B was hard-wired to 1.49(2) A, and C9B, C9D, and C13D were clamped to achieve roughly isotropic thermal vibrations using a SHELX-compatible ISOR command with 0.01 sigma and 0.05 sigma for the terminal atoms. All hydrogen atoms were placed in the calculated positions using the rider model with a fixed Uiso 1.2 for all CH and No. 7 groups. Largest peak: 0.76 AU A-3 at 0.1943 0.1800 0.0797 [0.42 A from S1B]. Deepest hole: -1.18 AU-3 at 0.2203 0.1543 0.1130 [0.86 A from S1B].

- 45 042432

Crystallographic data for C ₁₅ H9ClF ₃ N ₃ O ₂ S ₂ (M=419.82 g/mol): triclinic, space group P-1 (No. 2), a=10.0426(2) A, b=12.6946(3) A, c=13.5882(3) A, α=89.219(2)°, β=83.540(2)°, γ=73.357(2)°, V=1648.89(6) A3, Z=4, Т=120(1) K, μ (MoKa)=0.533 mm-1, Dcalc _C l=1.691 g/cm ³ , total

106625 reflections (6.04°<2Θ<62.5°), 10259 unique reflections (Rint=0.0431, Rsigma=0.0252) that were used for all calculations. Final R ₁ =0.0700 (>2sigma(I)) and wR ₂ =0.1358 (all data).

The single crystal structure of compound 1 is shown in Fig. 5A. In FIG. 5B shows the single crystal structure of the asymmetric cell of compound 1. The asymmetric cell was found to contain two independent molecules of compound 1 with an adjusted occupancy for the 1-chlorotrifluorophenyl moiety of the 'B' molecule of 34.1:43.2:22.7%. No other disordering was found for the entire model.

In table. 2 shows crystallographic refinement details for Compound 1 (Form 1).

table 2

Empirical Formula C1 ₅ H ₉ C1F3N3O2S2 Estimated mol.weight 419.82 Temperature / K 120(1) Crystal system triclinic space group P-1 a/a 10.0426(2) b/a 12.6946(3) s/a 13.5882(3) a/° 89.219(2) β/° 83.540(2) γ/° 73.357(2) Volume / A ³ 1648.89(6) Z, Z' 4 Rvichisl. g/cm 1.691 μ/mm ¹ 0.533 F(000) 848.0 Crystal size/mm ³ 0.237 x 0.158 x 0.126 Radiation ΜοΚα (λ = 0.71073) 2Θ Range for Data Acquisition/ ⁰ From 6.04 to 62.5 Index intervals -14<h< 14, -18<k< 17, -19<1< 19 Reflections received 106625 Independent reflections 10259 [Rim = 0.0431, R. _igma = 0.0252] Data / restrictions / parameters 10259/20/634 5 1.232 Final indices R [F ² > 2σ (F ² )] Ri = 0.0700, _wR2 = 0.1327 Final indices R [all data] Ri= 0.0787, wR ₂ = 0.1358 Armaks, Ar _MI n / A 0.76/-1.18 Ri = (Σ'F _o - F _c )/FF ₀ ) wR ₂ = {Σ [w(F ₀ ² - Fc ² ) ² ] /X[w(Fa ² ) ² }} ^m Ε=\Σ\ν (1Σ-1Σ ) ² ] / (u-p)} ^1/2

In table. 3 shows a simulated 20 powder X-ray diffraction (XRPD) plot of Compound 1 (Form 1). XRPD is shown in Fig. 6.

- 46 042432

Table 3

No. Position [°2Th.] FWHM [°2Th.] Area [imp*°2T11.] Interatomic distance d [A] Height [them.] Rel. inst. [%] 1 7.2633 0.096 89.50 12.1610 699.26 6.94 2 9.6858 0.096 119.57 9.1242 934.16 9.27 3 9.9875 0.120 158.90 8.8492 993.15 9.85 4 10.7073 0.120 101.23 8.2559 632.68 6.28 5 11.4394 0.096 122.72 7.7291 958.77 9.51 6 11.9245 0.096 32.83 7.4157 256.50 2.54 7 14.3055 0.096 317.49 6.1864 2480.38 24.61 8 14.5557 0.096 149.08 6.0806 1164.67 11.55 9 14.8445 0.096 233.35 5.9630 1823.05 18.09 10 15.1296 0.120 174.44 5.8512 1090.27 10.82 eleven 15.3639 0.096 111.84 5.7625 873.74 8.67 12 15.8539 0.096 381.04 5.5855 2976.84 29.53 13 16.5457 0.096 250.15 5.3535 1954.28 19.39 14 18.9653 0.096 125.47 4.6756 980.23 9.72 15 19.3893 0.168 842.79 4.5743 3762.45 37.32 16 19.7223 0.072 219.15 4.4978 2282.80 22.65 17 19.8198 0.072 244.04 4.4759 2542.05 25.22 18 20.3840 0.096 347.46 4.3533 2714.51 26.93 19 20.5478 0.072 98.72 4.3189 1028.31 10.20 20 20.9792 0.144 197.96 4.2311 1031.04 10.23 21 21.3094 0.072 304.21 4.1663 3168.82 31.44 22 21.5096 0.120 1522.85 4.1279 9517.79 94.42 23 21.8210 0.096 200.37 4.0697 1565.37 15.53 24 22.7420 0.120 92.33 3.9070 577.05 5.72 25 22.9527 0.096 94.20 3.8716 735.93 7.30 26 23.3841 0.120 94.20 3.8011 588.74 5.84 27 24.3296 0.096 171.62 3.6555 1340.75 13.30 28 24.8115 0.072 167.72 3.5856 1747.10 17.33 29 25.0252 0.096 1290.30 3.5554 10080.44 100.00 thirty 25.3797 0.096 374.34 3.5066 2924.51 29.01 31 25.5790 0.072 103.60 3.4797 1079.11 10.71 32 26.6520 0.072 79.62 3.3420 829.39 8.23 33 26.8121 0.072 175.89 3.3224 1832.21 18.18 34 26.9266 0.072 180.08 3.3085 1875.81 18.61 35 27.4026 0.096 73.50 3.2521 574.20 5.70 36 27.9857 0.096 64.48 3.1857 503.75 5.00 37 28.2013 0.096 130.86 3.1618 1022.32 10.14 38 28.5619 0.096 264.26 3.1227 2064.57 20.48 39 29.0172 0.072 59.44 3.0747 619.21 6.14 40 29.5813 0.096 119.64 3.0174 934.67 9.27 41 29.7121 0.096 186.01 3.0044 1453.19 14.42 42 29.9864 0.120 308.97 2.9775 1931.07 19.16 43 30.3430 0.120 243.43 2.9433 1521.46 15.09 44 31.8318 0.096 89.32 2.8090 697.82 6.92 45 31.9921 0.096 84.06 2.7953 656.71 6.51

46 32.5089 0.096 76.75 2.7520 599.61 5.95 47 33.9457 0.168 124.88 2.6388 557.50 5.53 48 35.9745 0.120 96.42 2.4945 602.63 5.98 49 38.0146 0.192 150.07 2.3651 586.21 5.82 50 38.9727 0.072 54.51 2.3092 567.82 5.63

Example 13 In vivo studies using the Line 61 mThy1alpha-synuclein transgenic mouse model.

Conducted several studies of the introduction of compound 1 in vivo on line 61 (L61) mThy1-αphasynuclein transgenic mouse model of Parkinson's disease (PD). The mThy1-alpha-synuclein transgenic mouse model overexpresses wild-type human ASYN in Thy-1 promoter studies (commonly referred to as line 61 transgenic mice; Rockenstein et al., 2002). These transgenic mice develop an intense accumulation of alpha-synuclein (ASYN) in areas relevant to Parkinson's disease (Rockenstein et al., 2002; Chesselet et al., 2012; Games et al., 2013), neurodegeneration, including degeneration of dopaminergic neurons, decrease in dopamine (DA) levels and loss of T-helpers in the striatum (Masliah et al., 2000; Lam. et al., 2011), as well as motor

- 47 042432 defects (Fleming et al., 2004). Male transgenic and non-transgenic mice from the same litter (3-3.5 months old) were used for all in vivo tests described in this text.

i) Effect of Compound 1 on ASYN pathology and a marker of neuroprotection and autophagy.

Alpha-synuclein (ASYN) is a neuronal protein whose dysregulation has been observed in the pathogenesis of PD. The effect of Compound 1 on alpha-synuclein aggregation was evaluated in L61 ASYN transgenic and non-transgenic mice in a 1 month test. L61 ASYN transgenic mice (total 36 mice, n=8-11 mice per group) were injected (ip) with 1, 5 or 10 mg/kg of Compound 1 or vehicle control solution (5% DMSO + 20% Cremphor EL + 0.9% saline) per day for 1 month. Non-transgenic mice (total 18 mice, n=8-11 mice per group) were used as controls and were injected daily (ip) with 10 mg/kg of compound 1 or vehicle control solution (5% DMSO+20% Cremphor EL+ 0.9% saline) per day for 1 month. At the end of one month, mice were sacrificed and immunohistochemical (IHC) determinations of total alpha-synuclein deposition, insoluble alpha-synuclein deposits (PK+ resistant), microtubule-associated protein 1A/1B-light chain 3 (LC3), and monomeric alpha-synuclein levels were performed. synuclein in brain tissue.

The results of the test with 1 month of administration show that compound 1 at dosages of 1, 5 and 10 mg/kg (ip once a day) has a beneficial effect, which includes a decrease in the cortical hippocampus and striatum of the level of monomeric, total and resistant to Proteinase K (insoluble) ASYN according to immunohistochemistry (IHC) and/or biochemical methods. These results indicate that Compound 1 promotes the clearance of alpha-synuclein (ASYN), a neuronal protein that is clearly dysregulated in the pathogenesis of PD. In addition to positive effects on ASYN neuropathology, administration of compound 1 increased microtubule-associated protein 1A/1B-light chain 3 (LC3), a marker of autophagy and neuroprotective mechanisms. Finally, administration of Compound 1 also provided functional improvements in the motor performance of L61 ASYN transgenic mice administered for 3 months.

In FIG. 7 shows quantitative staining of total alpha-synuclein in transverse sections of the cortex, hippocampus, and striatum of L61 ASYN transgenic and control mice after intraperitoneal administration of Compound 1 or vehicle for 1 month. In FIG. 8 shows IHC staining of total alpha-synuclein deposits in representative cross-sectional images of the neocortex, hippocampus, and striatum of L61 ASYN transgenic and control mice following intraperitoneal administration of Compound 1 or vehicle for 1 month. Quantification and IHC staining of total alpha-synuclein was performed according to known methods (Rockenstein et al., J. Neurosci. Res., 2002, 68(5):568-78; Tanji et al., Acta Neuropathol., 2010, 120, 145-154; Nuber et al., Brain., 2013, Feb. 136(Pt 2):412-32). In FIG. 7 shows that the administration of Compound 1 (1, 5 or 10 mg/kg per day, intraperitoneally, for 1 month) reduced the total level of ASYN in the neuropil (A) cortex, (B) hippocampus and (C) striatum of transgenic mice according to compared with the control group (which was injected with the carrier) according to the quantitative data of immunohistochemistry. As shown in FIG. 7, the decrease in the level of total alpha-synuclein in the cortex, hippocampus and striatum due to the administration of compound 1 is statistically significant. In particular, the data in Fig. 7A show that when administered daily at 1, 5, and 10 mg/kg, Compound 1 reduced cortical total alpha-synuclein levels by 13%, 32%, and 38%, respectively, compared to the control (vehicle-treated) group. This is also seen in FIG. 8, which shows the deposition of total alphasynuclein in representative cross-sectional images of the cortex, hippocampus, and striatum in the brain tissues of these mice. Coloring in Fig. 8 shows that Compound 1 has a beneficial effect in reducing total alpha-synuclein levels in the cortex, hippocampus and striatum.

In FIG. 9 shows quantitative staining of PK-resistant alpha-synuclein in transverse sections of the cortex, hippocampus and striatum of L61 ASYN transgenic and control mice after ip administration of Compound 1 or vehicle for 1 month. In FIG. 10 shows IHC staining of PK-resistant alpha-synuclein deposits in representative cross-sectional images of the neocortex, hippocampus, and striatum of L61 ASYN transgenic and control mice following intraperitoneal administration of Compound 1 or vehicle for 1 month. Quantification and IHC staining of PK-resistant alpha-synuclein was performed according to known methods (Rockenstein et al., J. Neurosci. Res., 2002, 68(5):568-78; Tanji et al., Acta Neuropathol., 2010, 120, 145-154; Nuber et al., Brain., 2013, Feb. 136(Pt 2):412-32). As shown in FIG. 9 and 10, administration of Compound 1 (1, 5, or 10 mg/kg per day ip for 1 month) also reduced deposits of insoluble alpha-synuclein (PK+ resistant) in (A) cortex, (B) hippocampus, and (C) striatum of transgenic mice. In FIG. 9 shows that the decrease in the level of PK-resistant alpha-synuclein in the cortex, hippocampus and striatum due to the administration of compound 1 is statistically significant. In particular, the data in Fig. 9A show that when administered daily at 5 and 10 mg/kg, Compound 1 reduced the level of PK-resistant aalpha-synuclein in the cortex by 37% and 36%, respectively, compared to the control (vehicle-treated) group.

- 48 042432

Coloring in Fig. 10 shows that Compound 1 has a beneficial effect on lowering

PK-resistant alpha-synuclein in the cortex, hippocampus and striatum.

In FIG. 11 shows that administration of Compound 1 (1, 5 or 10 mg/kg per day ip for 1 month) decreased the level of monomeric ASYN in the (A) cortex and (B) hippocampus in the cytosolic fraction of brain homogenates in L61 ASYN transgenic mice. The biochemical study was carried out using the ProteinSimple© western method. Briefly, samples were mixed with pre-calculated volumes of 0.1 x sample buffer and 5 x fluorescent master mix with a final sample concentration of 0.4 mg/ml in 10 µl of solution to optimize signal and reduce evaporation. Approximately 0.4 µl of sample was mixed with 2 µl of 5x fluorescent master mix and 7.8 µl of 0.1x sample buffer, vortexed, centrifuged, and heated at 95°C for 5 min. After rapid cooling, the samples, blocking reagent, wash buffer, primary antibodies, secondary antibodies and chemiluminescent substrate were dispersed in their intended wells in a plate obtained from the manufacturer (Kit#PS-MK14, ProteinSimple). After loading the tablet, automatic separation and immunodetection were performed according to the specified settings.

The Compass program (ProteinSimple, version 2.6.7) was used to generate a report that included the molecular weight, area, area percentage, and signal to noise ratio for each detected protein. Target protein data was normalized for beta-actin levels and further normalized between cartridges. Data are presented as means±SEM (standard error of the mean).

In FIG. 11 shows that when administered daily at 1, 5, or 10 mg/kg, Compound 1 lowers the level of monomeric ASYN in the cortex compared to a control group of L61 transgenic mice (which were injected with vehicle) in a statistically significant manner.

In FIG. 12 and 13 show that administration of compound 1 (1, 5, or 10 mg/kg per day ip for 1 month) increased the level of immunochemical staining of microtubule-associated protein 1A/1B-light chain 3 (LC3) in (A) cortex and (B) striatum but not in (B) hippocampus of transgenic mice.

ii) The effect of compound 1 on motor activity.

The effect of Compound 1 on motor impairment (grip strength) and neuroinflammation marker (translocator protein (18 kDa)) was evaluated in L61 ASYN transgenic and non-transgenic mice in a 3 month test.

Compound 1 was injected with L61 ASYN transgenic mice and non-transgenic mice in the control group (ip, 1 time per day) at doses of 5 and 10 mg/kg for 3 months (total 79 mice, n=14-17 mice per group). The carrier control solution consisted of a solution containing 5% DMSO + 20% Cremphor EL + 0.9% saline. Baseline grip strength in mice was determined prior to initiation of administration and then again determined after 70 days of vehicle or Compound 1 administration (5 or 10 mg/kg ip once daily).

As shown in FIG. 14, administration of Compound 1 (5 or 10 mg/kg per day ip once a day) for 3 months of testing had a beneficial effect on the transgenic dysmotility phenotype in L61 ASYN transgenic mice. At baseline, there was a statistically significant deficit in grip strength in transgenic mice compared to non-transgenic mice. Administration of Compound 1 (5 and 10 mg/kg) had a beneficial effect on grip force deficiency in L61 ASYN transgenic mice. After 70 days of administration, transgenic mice treated with compound 1 at 5 and 10 mg/kg had significantly higher grip strength than vehicle-treated control group of transgenic mice.

iii) Effect of compound 1 on the neuroinflammation marker TSPO.

Neuroinflammation is associated with increased expression of the 18-kDA translocator protein (TSPO), which is an inflammatory marker and is present in the mitochondria of activated microglia, astroglia, and macrophages (Crawshaw and Robertson, 2017). The effect of compound 1 on the level of translocator protein (18 kDa) (TSPO) was evaluated on L61 ASYN transgenic and non-transgenic mice in the study described above with 3 months of administration. At the end of the test, mice were sacrificed and immunofluorescence (IF) detection of TSPO in brain tissues was performed.

In FIG. 15 shows the levels of TSPO immunochemical staining on representative cross sections of mouse cortex. As shown in FIG. 15A and 15B, administration of compound 1 (5 and 10 mg/kg ip once a day) significantly reduced the level of TPSPO in L61 ASYN transgenic mice compared to the control group (which was administered only vehicle). In FIG. 15A shows representative immunochemical staining for TSPO in the L61 cortex of transgenic mice injected once daily with Compound 1, relative to the control group (which was injected with vehicle only). In FIG. 15B quantitatively shows TPSO staining from representative cross sections. Brain tissues were fixed by immersion in 4% paraformaldehyde (PFA), sectioned with a vibratome, and representative sections were examined for TSPO by standard immunofluorescence (IF) staining. Briefly, the right half of the brain was fixed in phosphate-buffered 4% PFA (pH 7.4) at 4°C for 48 h and then a series of 40 μm thick coronal sections were made using a vibratome. Sections were incubated in a free floating state overnight at 4°C. Immunochemical staining for TSPO was performed using knockout validated anti-TSPO rabbit monoclonal antibodies (1:500; abl99779; abcam, Temecula, CA, USA) pre-conjugated with an Alexa Fluor 488 secondary antibody. Immunochemical staining, visualization and analysis were performed on encoded sections from transgenic mice of line 61 and non-transgenic mice. Slides were visualized with an EVOS Auto FL imaging system (ThermoFisher Scientific, Waltham, MA, USA) with a 10x objective (EVOS PlanFL PH2 LWD; AMEP4681). The digitized images were analyzed using the Halo image analysis software package (Indica Labs, Corrales, NM, USA) by placing the ROI frame in the neocortex (standardized frame position for all images). A thresholding algorithm was created and then applied in the same way to all images to determine the percentage of immunostained ROI TSPO cortex. The analysis results were then exported for charting and statistical analysis.

The representative IF images in FIG. 15A show that when administered once a day at 5 or 10 mg/kg, Compound 1 had a beneficial effect in reducing the level of TSPO in the cortex, which is visually noticeable by a decrease in the intensity of IF staining. In addition, the quantification in FIG. 15B showed that Compound 1 at 5 or 10 mg/kg lowered TSPO levels in a statistically significant manner compared to control (vehicle-treated) mice.

iv) Effect of compound 1 on the neuroinflammation marker GFAP.

Neuroinflammation is also associated with increased expression of gliofibrillary acidic protein (GFAP) in activated astrocytes, which is triggered by a number of molecules, including pro-inflammatory mediators released from activated microglia (Saijo et al., 2009). Increased expression of gliofibrillary acidic protein (GFAP) accompanies astroglial activation and gliosis during neurodegeneration (Brahmachari et al., 2006). The effect of Compound 1 on GFAP expression was evaluated in L61 ASYN transgenic and non-transgenic mice in a 1 month study. After 30 days, mice were sacrificed and immunohistochemical detection of GFAP in brain tissues was performed.

In FIG. 16 shows representative GFAP immunostaining in sections containing hippocampus, L61 transgenic mice injected once daily with Compound 1 compared to controls (vehicle alone). In FIG. 17 shows the quantification of the above GFAP immunostaining on representative brain sections. Mice brain tissues were fixed (immersed in 4% PFA) and then sectioned 40 µm thick using a vibratome. Representative sections containing the hippocampus were examined for GFAP using standard immunohistochemical staining methods. General GFAP immunostaining procedures were as described in Rockenstein et al., J. Neurosci. Res., 2002, 68(5):568-78. Representative immunohistochemical (IHC) images in FIG. 16 show that when administered once a day at 5 or 10 mg/kg, Compound 1 has a beneficial effect by reducing the level of GFAP in the cortex, which is visually observed as a decrease in the intensity of IHC staining. In addition, the quantitative study in FIG. 17 shows that at 10 mg/kg, Compound 1 reduces cortical GFAP levels in a statistically significant manner.

v) Effect of compound 1 on the level of dopamine transporter (DAT) immunohistochemical staining.

In Parkinson's disease, uncontrolled neuroinflammation caused by synergistic activation of microglia and astrocytes contributes to increased death of DA neurons in the striatum during neurodegeneration.

In FIG. 18 shows representative immunohistochemical staining of dopamine transporter (DAT) in sections corresponding to the striatum of L61 transgenic mice after once daily injection of compound 1 in comparison with the control group (which was injected with vehicle). In FIG. 19 shows the quantification of the above DAT staining from the level corresponding to the sagittal slices containing the striatum and cortex as a reference. Brain tissues were fixed by immersion in 4% PFA and sectioned with a vibratome, and representative sections corresponding to the striatum and cerebellum were examined for DAT by IHC staining.

DAT immunohistochemical staining was performed using monoclonal antibodies (1:500; MAB369; Millipore, Temecula, CA) and biotinylated secondary antibodies (1:100; BA4000, Vector Labs) and analysis was performed on L61 encrypted sections of transgenic mice and non-transgenic mice. Slides were digitized using a Nanozoomer automatic high-resolution scanner (Hamamatsu Corp.). The digitized images were analyzed using the Halo image analysis software package (Indica Labs) by placing the ROI frame in the dorsal striatum and the other

- 50 042432 guyu - to a separate region of comparison in the brain (to normalize the DAT signal). A thresholding algorithm was created and then applied in the same way to all images to determine the average optical density of DAT immunostaining in each ROI. The results of the analysis were then exported for charting and statistical analysis, and the ratio of optical densities of DAT in the striatum / DAT in the cortex (region of comparison) was calculated for each subject.

The representative IF images in FIG. 18 show that when administered once daily at 5 or 10 mg/kg, Compound 1 has a beneficial effect on the recovery of DAT levels in the striatum, as visually seen in the increase in immunofluorescence intensity compared to control group L61 mice injected with vehicle alone. . DAT density was quantified by calculating the intensity of immunofluorescence in sections of the striatum relative to sections of the cerebellum, obtaining the striatal/comparison ratio. The quantification in FIG. 19 shows that Compound 1 at 10 mg/kg lowers GFAP levels in a statistically significant manner.

vi) Effect of compound 1 on neuroinflammation and beta-amyloid plaques.

Neuroinflammation is associated with increased expression of the 18-kDa translocator protein (TSPO), which is present in the mitochondria of activated microglia, astroglia, and macrophages (Crawshaw and Robertson 2017). The effect of Compound 1 on TSPO expression was evaluated in L41 APP transgenic and non-transgenic mice in a 1 month study. L41 APP transgenic mice (total 36 mice, n=8-11 mice per group) were injected (intraperitoneally) once a day with 5 mg/kg of compound 1 or vehicle as control (5% DMSO + 20% Cremphor EL + 0.9% saline) for 3 months. Non-transgenic mice (total 18 mice, n=8-11 mice per group) were used as controls and were injected (ip) once daily with 10 mg/kg of Compound 1 (data not shown) or vehicle as controls ( 5% DMSO + 20% Cremphor EL + 0.9% saline) for 1 month. Mice were sacrificed 30 days later and immunofluorescent (IF) detection of TSPO in brain tissues was performed.

In FIG. 20 shows a quantitative analysis of TPSO staining in representative brain sections. Brain tissues were fixed by immersion in 4% PFA and sectioned with a vibratome and representative sections corresponding to the cortical neuropil were examined for TSPO using standard immunofluorescence (IF) staining. The results show that when administered once a day at a dosage of 5 mg/kg, compound 1 has a beneficial effect, reducing the level of TSPO in the cortex, which is visually noticeable by a decrease in the intensity of IF staining. In addition, the quantitative analysis in Fig. 20 shows that when administered once daily at 5 mg/kg, Compound 1 reduces TSPO levels in a statistically significant manner compared to 61 mice treated with vehicle alone.

vii) Effect of compound 1 on beta-amyloid plaques.

As previously described, 61 transgenic mice express mutant hAPP751 extensively and develop mature plaques in the cortex, hippocampus, thalamus, and olfactory region of the mouse brain. The effect of Compound 1 on beta-amyloid plaque formation was studied in L41 APP transgenic and non-transgenic mice in a 1 month study. After 30 days, mice were sacrificed and immunofluorescent (IF) detection of beta-amyloid in brain tissues was performed.

In FIG. 21 shows a quantitative analysis of beta-amyloid staining in L41 transgenic mice injected once a day with compound 1, relative to the control group (which was injected with the vehicle). Brain tissues were fixed by immersion in 4% PFA and sectioned with a vibratome and representative sections and neuropil-containing cortex, hypothalamus, and striatum were examined for beta-amyloid using standard immunohistochemical (IHC) staining.

Around day 30, all mice were sacrificed within 2 hours of the last injection and brains and other samples were taken. The brain was removed and divided in the sagittal plane. The right half of the brain was fixed in phosphate-buffered 4% PFA (pH 7.4) at 4°C for 48 h for neuropathological analysis. A series of coronal sections 40 µm thick were then made from the immersion-fixed halves of the brain using a vibratome. Sections were incubated in a free-floating state overnight at 4°C with primary antibodies. To confirm the specificity of primary antibodies, control experiments were performed in which sections were incubated overnight in the absence of primary antibodies (removed), non-immune serum, or primary antibodies preadsorbed for 48 hours with a 20-fold excess of the corresponding peptide.

An immunostaining study of β-amyloid pathology was performed with purified antibodies to β-amyloid 1-16 (1:500; clone 6E10, reactive with amino acid residues 1-16 of P-amyloid and APP; #SIG-39320; Covance Research Products, Inc. ., Dedham, MA, USA). After incubation with primary antibodies, sections were then incubated with biotinylated secondary antibodies (1:200, Vector Laboratories, Burlingame, CA) and visualized using avidin-

Claims (49)

biotin (ABC) kit (Vector Laboratories, Burlingame, CA) with diaminobenzidine tetrahydrochloride (DAB; Sigma-Aldrich, St. Louis, MO) as chromogen. The resulting slides were visualized at 40x magnification using the Hamamatsu Nanozoomer™ high resolution scanner located at the Microscopy Core of the UCSD Department of Microbiology. The digitized images were then transferred to Neuropore and analyzed using the Halo® software package (Indica Labs, Corrales, NM). The same standardized area mask (region of interest (ROI) with the same dimensions for the analysis of the same area) was superimposed on each image and placed over the dorsal striatum. The window for comparison was determined using representative images in the control group (which was injected with the vehicle), saved and then applied to all images using a batch processing algorithm. Results are shown as percentage (%) of ROI area immunopositive for each marker. The images were processed to correct sample and image problems and any identified problems were corrected prior to sample disclosure and statistical analysis. The results obtained show that when administered once a day at a dosage of 5 mg/kg, compound 1 has a beneficial effect by lowering the level of beta-amyloid in the cortex, which is visually determined by the decrease in the intensity of IF staining. In addition, the quantification in FIG. 20 shows that when administered once a day at a dosage of 5 mg/kg, compound 1 lowers the level of beta-amyloid in a statistically significant manner in comparison with the control group of mice of line 41, which were injected with vehicle only. For all figures, all data are given as group means ± standard error of the mean (****p<0.0001 or *p<0.05 means statistically significant baseline phenotype or phenotype in the control group (vehicle) compared to the control group (vehicle). ) non-transgenic mice; #p<0.05, ##p<0.01, ###p<0.001 or ####p<0.0001 indicates a statistically significant effect of administration in transgenic groups on compound 1 compared to control transgenic groups (which were administered only carrier)). CLAIM 1. Compound having formula (I) where R ¹ , R ² and R ³ each independently represents a hydrogen atom, a hydroxy group, a halogen, an unsubstituted C1-4-alkyl, a C1-4-alkyl substituted with one or more substituents selected from the group consisting of hydroxyl, halogen , -NR ^f R ^g , cyano groups, nitro groups, C 1-4 alkoxy groups and C _1-4 haloalkoxy groups, where R ^f and R ^g are each independently H or C 1-4 alkyl, an unsubstituted C1-4 alkoxy group, a C1-4 alkoxy group substituted with hydroxyl, halogen, -NR ^f R ^g , a cyano group, a nitro group, a C1-4 alkoxy group, and a C _1-4 haloalkoxy group -group, where R ^f and R ^g each independently represents H or C _1-4 -alkyl, -CN, -C(O)R ^x , -C(O)OR ^x , -S(O)2R ^x or - NR ^y R ^z ; R ^x , R ^y and R ^z are each independently H, unsubstituted C1-4 alkyl or C1-4 alkyl substituted with one or more hydroxyls, or R ^y and R ^z together with the nitrogen atom to which they are attached form an unsubstituted monocyclic heterocycloalkyl ring or a monocyclic heterocycloalkyl ring substituted with one or more substituents selected from the group consisting of C1- _C6 alkyl, _C2 - _C6 alkenyl, _C2 - _C6 alkynyl, hydroxyl, halogen, -NR ^f R ^g , cyano groups, nitro groups, C1-4 alkoxy groups and C1-4 haloalkoxy groups, where R ^f and R ^g are each independently H or C1-4 alkyl and where the monocyclic heterocycloalkyl ring is a monocyclic 5- or 6-membered heterocyclic ring containing 1 or 2 heteroatoms selected from nitrogen and oxygen; or a pharmaceutically acceptable salt thereof. 2. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R ¹ is a hydrogen atom, an unsubstituted C _1-4 alkoxy group, a substituted C _1-4 alkoxy group, or -NR ^y R ^z . 3. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ¹ is a hydrogen atom. 4. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ¹ is a C 1 _-4 alkoxy group. 5. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ¹ is -OCH2CH2-O-CH2CH3 or -OCH2CH2OCH3. 6. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ¹ is -NR ^y R ^z , where R ^y and R ^z are each independently H or C _1-4 alkyl. 7. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R ¹ is -NHCH2CH2OH or -N(CH2CH3)2. - 52 042432 8. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, where R ¹ is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form a substituted or unsubstituted monocyclic heterocycloalkyl ring. 9. The compound according to claim 8, or a pharmaceutically acceptable salt thereof, wherein R ¹ is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form a monocyclic heterocycloalkyl ring selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl, wherein morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl are each unsubstituted or substituted. 10. A compound according to claim 8 or 9, or a pharmaceutically acceptable salt thereof, wherein R ¹ is morpholinyl, 4-methyl-piperazin-1-yl, piperidinyl, or pyrrolidinyl. 11. A compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein R ² is a hydrogen atom, C ₁ . ₄ -alkyl or substituted C ₁ . ₄ -alkyl. 12. A compound according to claim 11, or a pharmaceutically acceptable salt thereof, wherein R ² is C ₁ . ₄ -alkyl substituted with halogen. 13. A compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein R ² is CF3. 14. A compound according to claim 11, or a pharmaceutically acceptable salt thereof, wherein R ² is methyl. 15. A compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein R ² is a substituted or unsubstituted C ₁ . _4- alkoxy group, -CN or -NR ^y R ^z . 16. The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein R ² is -NR ^y R ^z , where R ^y and R ^z are each independently H or C ₁ . ₄ -alkyl, where C ₁ . ₄ -alkyl unsubstituted or substituted. 17. A compound according to claim 16, or a pharmaceutically acceptable salt thereof, wherein R ² is -N(CH3)2. 18. The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein R ² is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form a substituted or unsubstituted monocyclic heterocycloalkyl ring. 19. The compound according to claim 18, or a pharmaceutically acceptable salt thereof, wherein R ² is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form a monocyclic heterocycloalkyl ring selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl, wherein morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl are each unsubstituted or substituted. 20. A compound according to any one of claims 15, 18 or 19, or a pharmaceutically acceptable salt thereof, wherein R ² is morpholinyl. 21. A compound according to claim 15, or a pharmaceutically acceptable salt thereof, wherein R ² is -CN. 22. A compound according to claim 15, or a pharmaceutically acceptable salt thereof, wherein R ² is C ₁ . ₄ alkoxy group, unsubstituted or substituted. 23. A compound according to claim 15, or a pharmaceutically acceptable salt thereof, wherein R ² is a methoxy group, -OCH ₂ CH ₂ -O-CH ₂ CH ₃ or -OCH ₂ CH ₂ OCH ₃ . 24. A compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R ³ is halogen. 25. A compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R ³ is chlorine. 26. A compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R ³ is a hydrogen atom. 27. A compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R ³ is C ₁ . ₄ -alkyl or substituted C ₁ . ₄ -alkyl. 28. A compound according to claim 27, or a pharmaceutically acceptable salt thereof, wherein R ³ is methyl. 29. A compound according to claim 27, or a pharmaceutically acceptable salt thereof, wherein R ³ is C ₁ . ₄ -alkyl substituted with one or more halogen atoms. 30. A compound according to claim 29, or a pharmaceutically acceptable salt thereof, wherein R ³ is CF3. 31. A compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R ³ is -CN. 32. A compound according to any one of claims 1-23, or a pharmaceutically acceptable salt thereof, wherein R ³ is -NR ^y R ^z , where R ^y and R ^z are each independently H or C ₁ . ₄ -alkyl, where C ₁ . ₄ -alkyl unsubstituted or substituted. 33. The compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, wherein R ³ is -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form a substituted or unsubstituted monocyclic heterocycloalkyl ring . 34. The compound according to claim 33, or a pharmaceutically acceptable salt thereof, where R ³ is - 53 042432 -NR ^y R ^z and R ^y and R ^z together with the nitrogen atom to which they are attached form a monocyclic heterocycloalkyl ring selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl, where morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl are each unsubstituted or substituted. 35. A compound according to claim 33 or 34, or a pharmaceutically acceptable salt thereof, wherein R ³ is morpholinyl. 36. A compound according to any one of claims 1-23, or a pharmaceutically acceptable salt thereof, wherein R ³ is a C _1-4 -alkoxy group, unsubstituted or substituted. 37. A compound selected from the group consisting of the following compounds: Compound Structure Name 1 Οχ /O ¥ X xx J 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione; 2 Οχ /О W 0 ' 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-morpholinophenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione; 3 OO oV 4 NH 4-(4-(phenylsulfonyl)phenyl)-2,4-dihydro3H-1,2,4-triazole-3-thione; 4 I "=/ 'z N V o 4-(4-((4-chlorophenyl)sulfonyl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione; 5 XX xx J 4-(4-((4-chloro-3-methylphenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3 thione; 6 About 0 VOU 4NH 4-((4-(5-thioxo-1,5-dihydro-4H-1,2,4triazol-4-yl)phenyl)sulfonyl)benzonitrile; 7 0 V 0° z I 4-(4-((4-morpholinophenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3 thione; - 54 042432 8 Ο _χ 0 Guide ^and । NH 4-(4-((4-methoxyphenyl)sulfonyl)phenyl)2,4-dihydro-3H-1,2,4-triazole-3-thione; 9 7 NH H' 4-(4-tosylphenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione; 10 to 0 I 4-(4-((4-fluorophenyl)sulfonyl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione; eleven T o 0... 0° t 4-(4-((3-(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione; 12 oh oh 00 01 l mi । NH 4-(4-((3-methoxyphenyl)sulfonyl)phenyl)2,4-dihydro-3H-1,2,4-triazole-3-thione; 13 I 'ζ .0 >ω U o s 4-(4-((3-(2-ethoxyethoxy)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione; 14 0.0xV/ ΥΎ Υ0 l ^Γνη ^Ν 3-((4-(5-thioxo-1,5-dihydro-4H-1,2,4triazol-4-yl)phenyl)sulfonyl)benzonitrile; 15 1 o o 1 Λ// ish 4NH 4-(4-((3-(dimethylamino)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione; - 55 042432 16 ο. ,ο H NH 4-(4-((3-morpholinophenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4-triazole-3 thione; 17 About 0w. 4-(4-(m-tolylsulfonyl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione; 18 °\P W HN ^ _N ' ΌΗ -1,2,4-triazole-3 thione; 19 PP O - 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-(piperidin-1-yl)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione ; 20 PP\\// 0' 1 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-(4-methylpiperazin-1 yl)phenyl)-2,4-dihydro-3H-1,2,4-triazole- 3tion; 21 PP P - 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-(diethylamino)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione; 22 PPX% 4-(4-((4-chloro-3-(trifluoromethyl)phenyl)sulfonyl)-2-(2-ethoxyethoxy)phenyl)-2,4dihydro-3H-1,2,4-triazole-3-thione; 23 ΤΊ tL 0° I 4-(4-((4-methyl-3-(trifluoromethyl)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazole-3-thione; ²⁴ P o \\// 'Ppp 0 4 1,2,4-triazole-3-thione; and - 56 042432 4-(4-((3-(2-methoxyethoxy)phenyl)sulfonyl)phenyl)-2,4-dihydro-3H-1,2,4triazol-3-thione, or a pharmaceutically acceptable salt thereof. 38. A compound having the following structure: or a pharmaceutically acceptable salt thereof. 39. A compound having the following structure: or a pharmaceutically acceptable salt thereof. 40. A pharmaceutical composition for the treatment of a pathological condition caused by neurodegeneration or protein accumulation, comprising (a) at least one compound according to any one of claims 1 to 39 or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable excipient. 41. The pharmaceutical composition of claim 40, wherein the pharmaceutically acceptable excipient is a polymeric agent. 42. The pharmaceutical composition according to claim 40, wherein the pharmaceutically acceptable excipient is selected from the group consisting of carboxymethyl cellulose (CMC), hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC), hydroxypropyl methyl cellulose (HPMC), gelatin, gelatin hydrolyzate, sucrose, dextrose, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), vinylpyrrolidone copolymers, pregelatinized starch, sorbitol and glucose; and polyacrylates. 43. The pharmaceutical composition according to claim 40, wherein the pharmaceutically acceptable excipient is selected from the group consisting of hydroxypropyl methylcellulose (HPMC), polyvinylpyrrolidone (PVP), and collidone. 44. The pharmaceutical composition according to any one of claims 40 to 43, wherein the pharmaceutical composition is in the form of a dry spray dispersion (SDD). 45. A method of treating a pathological condition caused by neurodegeneration or protein accumulation, comprising administering to a subject in need of such treatment an effective amount of at least one compound according to any one of claims 1 to 39 or a pharmaceutically acceptable salt thereof, wherein said pathological condition is a disease Alzheimer's disease, Parkinson's disease, Lewy body dementia, Parkinson's disease dementia, multiple system atrophy, amyotrophic lateral sclerosis, or progressive supranuclear palsy. 46. The use of a compound according to any one of claims 1 to 39, or a pharmaceutically acceptable salt thereof, for the treatment of a pathological condition caused by neurodegeneration or protein accumulation, where said pathological condition is Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, dementia due to Parkinson's disease, multiple system atrophy, amyotrophic lateral sclerosis or progressive supranuclear palsy. 47. The use of at least one compound according to any one of claims 1 to 39 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a pathological condition caused by neurodegeneration or accumulation of proteins, where said pathological condition is Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, dementia due to Parkinson's disease, multiple system atrophy, amyotrophic lateral sclerosis, or progressive supranuclear palsy. 48. A method for preventing aggregation, or accumulation, or improving the clearance of a protease-resistant protein, comprising contacting the protease-resistant protein with an effective amount of at least one compound according to any one of claims 1-39, or a pharmaceutically acceptable salt thereof, where the specified contact occurs in vitro, ex vivo or in vivo. 49. The method of claim 48, wherein the protease resistant protein is selected from alpha-synuclein, -