Compounds, compositions and methods for treating conditions associated with protein misfolding

Abstract

The present disclosure provides compounds of Formula (I) and Formula (II) useful for treating or reducing the severity of a disease or condition, including those associated with alpha-synuclein.

Description

Background Technology

[0001] Alpha-synuclein (α-syn) is a 140-amino acid protein encoded by the SNCA gene and is one of the most abundant proteins expressed in the nervous system (accounting for 1% of total cytoplasmic proteins). It is essentially disordered in aqueous solution but forms α-helical structures upon binding to negatively charged lipids. Under physiological conditions, it is primarily found in neurons and located at the presynaptic terminals. Its physiological functions are only partially understood; the protein is thought to participate in membrane interactions (particularly mitochondria) and play a role in regulating dopamine levels. Aggregated forms of α-syn (“Lewy bodies” and “Lewy neurites”) are hallmarks of Parkinson’s disease (PD), Lewy body dementia (DLB), and multiple system atrophy (MSA). Extensive preclinical data suggest that smaller, soluble aggregates of α-syn (called “oligomers”) are key drivers of neurotoxicity. These oligomeric forms of α-synuclein have been shown to possess broad neurotoxicity and are a cause of the pathogenesis and progression of Parkinson’s disease (PD). These oligomers bind to membranes, receptors, and organelles, disrupting metabolic and neuronal functional pathways and ultimately leading to neuronal death. Summary of the Invention

[0002] This disclosure includes compounds of formula (I) or formula (II).

[0003]

[0004] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3 and R 4 Defined in this article.

[0005] Furthermore, this disclosure includes methods for preparing compounds of formula (I) or formula (II) and methods for using compounds of formula (I) or formula (II). Detailed Implementation

[0006] definition

[0007] As used herein, the term "aliphatic" or "aliphatic group" means a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon (also referred to herein as "carbocyclic," "cycloaliphatic," or "cycloalkyl") that is fully saturated or contains one or more unsaturated units but is not aromatic and has a single connection point to the rest of the molecule. Unless otherwise stated, an aliphatic group contains 1 to 6 aliphatic carbon atoms. In some embodiments, the aliphatic group contains 1 to 5 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1 to 4 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1 to 3 aliphatic carbon atoms, and in yet another embodiment, the aliphatic group contains 1 to 2 aliphatic carbon atoms. In some embodiments, "cycloaliphatic" (or "carbocyclic" or "cycloalkyl") means a monocyclic C3-C6 hydrocarbon that is fully saturated or contains one or more unsaturated units but is not aromatic and has a single connection point to the rest of the molecule. Suitable aliphatic groups include, but are not limited to, straight-chain or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and their hybrids, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0008] The term "halogenated aliphatic" refers to an aliphatic group that has been replaced by one or more halogen atoms.

[0009] The term "alkyl" refers to a straight-chain or branched alkyl group. Exemplary alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tert-butyl.

[0010] The term "halogenated alkyl" refers to a straight-chain or branched alkyl group that has been substituted with one or more halogen atoms.

[0011] The term "halogen" refers to F, Cl, Br, or I.

[0012] The term "aryl," used alone or as part of a larger portion of terms such as "aralkyl," "aralkyloxy," or "aryloxyalkyl," refers to a monocyclic or bicyclic system having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. The term "aryl" is used interchangeably with the term "aryl ring." In certain embodiments of this disclosure, "aryl" refers to an aromatic ring system that may have one or more substituents, including but not limited to phenyl, biphenyl, naphthyl, anthracene, etc. As used herein, the scope of the term "aryl" also includes groups in which the aromatic ring is fused with one or more non-aromatic rings, such as indanyl, phthalimide, naphthimide, phenanthridine, or tetrahydronaphthyl.

[0013] The terms "heteroaryl" and "heteroary-" used alone or as part of a larger portion (e.g., "heteroarylalkyl" or "heteroarylalkoxy") refer to a group having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; sharing 6, 10, or 14 π electrons in a cyclic array; and having one to five heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, as well as any quaternized form of basic nitrogen. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrroleyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiazolyl, pyridinyl, pyrazinyl, indoleazinyl, purine, naphthidyl, and pteridinyl. As used herein, the terms "heteroaryl" and "heteroary-" also include groups in which a heteroaryl ring is fused with one or more aryl, cycloaliphatic, or heterocyclic rings, wherein the group or connecting point is on the heteroaryl ring. Non-limiting examples include indolyl, isoindolyl, benzothiopheneyl, benzofuranyl, dibenzofuranyl, indazoleyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, terpineyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolinyl, carbazolyl, acridineyl, phenazinyl, phenothiazinyl, phenotoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic or bicyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl," or "heteroarylene," any of which includes an optionally substituted ring. The term "heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl portions are optionally substituted independently.

[0014] As used herein, the terms “heterocycle,” “heterocyclic group,” “heterocyclic group,” and “heterocyclic ring” are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic portion that is saturated or partially unsaturated and has one or more, preferably one to four, heteroatoms as defined above, in addition to a carbon atom. When referring to the ring atom of a heterocycle, the term “nitrogen” includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, or nitrogen, nitrogen may be N (e.g., in 3,4-dihydro-2H-pyrrole), NH (e.g., in pyrrolealkyl), or... +NR (as in TV-substituted pyrrolidinyl groups). Heterocycles can be attached to their side groups at any heteroatom or carbon atom that produces a stable structure, and any one of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, azolealkyl, piperazineyl, dioxacyclohexyl, dioxacyclopentyl, diazacycloheptenyl, oxazocycloheptenyl, thiocycloheptenyl, morpholinyl, and quininecycloyl. The terms “heterocyclic,” “heterocyclic group,” “heterocyclic ring,” “heterocyclic moiety,” and “heterocyclic group” are used interchangeably herein and also include groups in which the heterocyclic ring is fused with one or more aryl, heteroaryl, or cycloaliphatic rings, such as indololinyl, 3H-indolyl, benzodihydropyranyl, phenanthridineyl, or tetrahydroquinolinyl, wherein the group or connecting point is on the heterocyclic ring. The heterocyclic group can be monocyclic or bicyclic. The term “heterocyclic alkyl” refers to an alkyl group substituted with a heterocyclic group, wherein the alkyl group and the heterocyclic moiety are optionally substituted independently.

[0015] As used herein, the term "partially unsaturated" refers to a ring moiety containing at least one double or triple bond. The term "partially unsaturated" is intended to cover rings having multiple unsaturated sites, but is not intended to include aryl or heteroaryl moiety as defined herein.

[0016] As described herein, the compounds of this disclosure may contain an "optionally substituted" portion. Generally, regardless of whether the term "optionally" is preceding it, the term "substituted" means that one or more hydrogens of the specified portion are replaced by suitable substituents. Unless otherwise indicated, the "optionally substituted" group may have suitable substituents at each substituted position of the group, and the substituents at each position may be the same or different when more than one position in any given structure may be substituted by more than one substituent selected from the specified group. The combinations of substituents contemplated by this disclosure are preferably those that result in the formation of stable or chemically viable compounds. As used herein, the term "stable" means a compound that remains substantially unchanged when subjected to conditions permissible for the production, detection, and, in some embodiments, recovery, purification, and use for one or more purposes disclosed herein.

[0017] The suitable monovalent substituent on the substituted carbon atom of the "optionally substituted" group is independently a halogen; —(CH2) 0-4 R ° ;—(CH2) 0-4 OR ° —O(CH2) 0-4 R ° —O—(CH2) 0-4 C(O)OR ° ;—(CH2)0-4 CH(OR ° )2;—(CH2) 0-4 SR ° ;—(CH2) 0-4 Ph, which can be R ° Substitution; —(CH2) 0-4 O(CH2) 0-1 Ph, which can be R ° Replacement; —CH═CHPh, which can be replaced by R ° Substitution; —(CH2) 0-4 O(CH2) 0-1 -pyridyl group, which can be R ° Substitution; —NO2; —CN; —N3; —(CH2) 0-4 N(R ° )2;—(CH2) 0-4 N(R ° )C(O)R ° ;—N(R ° )C(S)R ° ;—(CH2) 0-4 N(R ° )C(O)NR ° 2; —N(R) ° )C(S)NR ° 2; —(CH2) 0-4 N(R ° )C(O)OR ° ;—N(R ° )N(R ° )C(O)R ° ;—N(R ° )N(R ° )C(O)NR ° 2; —N(R) ° )N(R ° )C(O)OR ° ;—(CH2) 0-4 C(O)R ° ;—C(S)R ° ;—(CH2) 0-4 C(O)OR ° ;—(CH2) 0-4 C(O)SR ° ;—(CH2) 0-4 C(O)OSiR ° 3; —(CH2) 0-4 OC(O)R ° ;—OC(O)(CH2) 0-4 SR ° SC(S)SR °;—(CH2) 0-4 SC(O)R ° ;—(CH2) 0-4 C(O)NR ° 2; —C(S)NR ° 2; —C(S)SR ° ;—SC(S)SR ° —(CH2) 0-4 OC(O)NR ° 2; —C(O)N(OR) ° )R ° ;—C(O)C(O)R ° ;—C(O)CH2C(O)R ° ;—C(NOR ° )R ° ;—(CH2) 0-4 SSR ° ;—(CH2) 0-4 S(O)2R ° ;—(CH2) 0-4 S(O)2OR ° ;—(CH2) 0-4 OS(O)2R ° ;—S(O)2NR ° 2; —(CH2) 0-4 S(O)R ° ;—N(R ° )S(O)2NR ° 2; —N(R) ° )S(O)2R ° ; —N(OR ° )R ° ;—C(NH)NR ° 2; —P(O)2R ° ;—P(O)R ° 2; —OP(O)R ° 2; —OP(O)(OR ° )2; SiR ° 3; —(C 1-4 (straight-chain or branched alkylene)O—N(R) ° )2; or —(C 1-4 (straight-chain or branched alkylene)C(O)O—N(R) ° )2, where each R ° It can be substituted as defined below and independently be hydrogen, C 1-6 Aliphatic, —CH2Ph, —O(CH2) 0-1Ph, —CH2- (5-6 membered heteroaryl ring), or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 independent heteroatoms selected from nitrogen, oxygen, or sulfur, or, although defined above, two independently occurring R... ° Together with one or more of its intervening atoms, it forms a 3-12 saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 independent heteroatoms selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

[0018] R ° (or by using two independently occurring R) ° The suitable monovalent substituents on the ring (formed together with its intercalary atoms) are independently halogens, —(CH2). 0-2 R ● -(halogenated R) ● ), —(CH2) 0-2 OH, —(CH2) 0-2 OR ● —(CH2) 0-2 CH(OR ● )2;—O(halogenated R ● —CN, —N3, —(CH2) 0-2 C(O)R ● —(CH2) 0-2 C(O)OH、—(CH2) 0-2 C(O)OR ● —(CH2) 0-2 SR ● —(CH2) 0-2 SH、—(CH2) 0-2 NH2、—(CH2) 0-2 NHR ● —(CH2) 0-2 NR ● 2、—NO2、—SiR ● 3. —OSiR ● 3、—C(O)SR ● —(C 1-4 (straight-chain or branched alkylene)C(O)OR ● Or —SSR ● , where each R ● For unsubstituted or halogenated sites, only one or more halogens are substituted, and the halogens are independently selected from C. 1-4 Aliphatic, —CH2Ph, —O(CH2) 0-1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 independent heteroatoms selected from nitrogen, oxygen, or sulfur. R °Suitable divalent substituents on saturated carbon atoms include =O and =S.

[0019] Suitable divalent substituents on the saturated carbon atom of the "optionally substituted" group include the following: ═O, ═S, ═NNR*2, ═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O)2R*, ═NR*, ═NOR*, —O(C(R*2)) 2-3 O—or —S(C(R*2)) 2-3 S—, where each independently occurring R* is selected from hydrogen, and the substituted C can be defined as follows. 1-6 Aliphatic groups, or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl rings having 0-4 independent heteroatoms selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents bonded to the ortho-substituted carbon of the "optionally substituted" group include: —O(CR*2). 2-3 O—, where each individually occurring R* is selected from hydrogen, and C can be substituted as defined below. 1-6 Aliphatic, or unsubstituted 5-6 saturated, partially unsaturated or aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0020] Suitable substituents on the aliphatic group of R* include halogens, -R ● -(halogenated R) ● —OH, —OR ● —O(halogenated R) ● ), —CN, —C(O)OH, —C(O)OR ● —NH2, —NHR ● —NR ● 2 or —NO2, where each R ● For unsubstituted or halogenated sites, only one or more halogens are substituted, and each site is independently C. 1-4 Aliphatic, —CH2Ph, —O(CH2) 0-1 Ph, or a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0021] Suitable substituents on the substituted nitrogen of the "optionally substituted" group include —R † —NR † 2、—C(O)R † —C(O)OR † —C(O)C(O)R † —C(O)CH2C(O)R † —S(O)2R † —S(O)2NR † 2、—C(S)NR †2、—C(NH)NR † 2 or —N(R) † )S(O)2R † ; where each R † C that can be substituted independently for hydrogen, as defined below. 1-6 Aliphatic, unsubstituted -OPh, or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl rings having 0-4 independently selected heteroatoms chosen from nitrogen, oxygen, or sulfur, or, despite the above definition, two independently occurring R... † Together with its intercalary atoms, it forms an unsubstituted 3-12 saturated, partially unsaturated, or aryl monocyclic or bicyclic ring with 0-4 independent heteroatoms selected from nitrogen, oxygen, or sulfur.

[0022] R † Suitable substituents on the aliphatic group can be halogens, -R ● -(halogenated R) ● —OH, —OR ● —O(halogenated R) ● ), —CN, —C(O)OH, —C(O)OR ● —NH2, —NHR ● —NR ● 2 or —NO2, where each R ● For unsubstituted or halogenated sites, only one or more halogens are substituted, and each site is independently C. 1-4 Aliphatic, —CH2Ph, —O(CH2) 0-1 Ph, or a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0023] As used herein, the term "pharmaceutically acceptable salt" means that salts suitable for contact with tissues of humans and lower animals without undue toxicity, irritation, allergic reactions, etc., within the bounds of reasonable medical judgment and in proportion to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, which is incorporated herein by reference. Pharmaceutically acceptable salts of the compounds disclosed herein include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed by reacting amino groups with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, disglucuronate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-heptahydrate, glyceryl phosphate, gluconate, hemisulfate, heptahydrate, hexanoate, hydroiodate, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, dodecyl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc.

[0024] Salts derived from suitable bases include alkali metals, alkaline earth metals, ammonium, and N(C) salts. 1-4 Alkyl)4 salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Where appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate ions.

[0025] The combinations of substituents and variables contemplated in this disclosure are only those that result in the formation of stable compounds. As used herein, the term "stable" means a compound that has sufficient stability to allow manufacture and maintains the integrity of the compound for a period of time sufficient for its use for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0026] The description of the list of chemical groups in any definition of a variable herein includes the definition of the variable as any single group or combination of the listed groups. The description of embodiments of a variable herein includes the embodiment as any single embodiment or combination with any other embodiment or part thereof.

[0027] As used herein, the term "biological sample" includes, but is not limited to, cell cultures or extracts thereof; biopsy material or extracts thereof obtained from mammals; and blood, saliva, urine, feces, semen, tears or other bodily fluids or extracts thereof. Examples of such purposes include, but are not limited to, blood transfusions, organ transplantation, biological sample storage, and bioassays.

[0028] As used herein, "therapeutic effective amount" means the amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that elicits a desired biological response. In some embodiments, a therapeutically effective amount of a substance is an amount sufficient to treat, diagnose, prevent, and / or delay the onset of a disease, condition, and / or disorder when administered as part of a dosing regimen to a subject who has or is susceptible to the disease, condition, and / or disorder. As will be understood by those skilled in the art, the effective amount of a substance can vary depending on factors such as the desired biological endpoint, the substance to be delivered, the target cells, or tissues. For example, an effective amount of a compound provided in a formulation for treating a disease, condition, and / or disorder is an amount that reduces, improves, alleviates, inhibits, prevents, delays the onset of one or more symptoms or features of the disease, condition, and / or disorder, reduces its severity, and / or reduces its incidence.

[0029] As used herein, the term "treatment" means to partially or completely reduce, suppress, or delay the onset of a condition or disorder as described herein, or one or more symptoms of such condition or disorder, or to prevent, improve, and / or alleviate such condition or disorder, or one or more symptoms of such condition or disorder. In some embodiments, treatment may be administered after one or more symptoms have developed. In some embodiments, the term "treatment" includes preventing or halting the progression of a disease or disorder. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to susceptible individuals before the onset of symptoms (e.g., based on a history of symptoms and / or based on genetic or other susceptibility factors). Treatment may also continue after symptoms have subsided, for example, to prevent or delay their recurrence. Therefore, in some embodiments, the term "treatment" includes preventing the recurrence or relapse of a disease or disorder.

[0030] As used herein, the term "patient" refers to an animal, preferably a mammal, and most preferably a human.

[0031] The term "pharmaceutically acceptable carrier, adjuvant, or mediator" refers to a non-toxic carrier, adjuvant, or mediator that does not impair the pharmacological activity of the compound formulated with it. Pharmaceutically acceptable carriers, adjuvants, or mediators that may be used in compositions of the compounds disclosed herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of saturated vegetable fatty acids in the form of glycerides, water, salts, or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin.

[0032] "Pharmaceutically acceptable derivative" means any non-toxic salt, ester, salt of ester or other derivative of the compounds disclosed herein, which, upon administration to a recipient, can directly or indirectly provide the compounds disclosed herein or their inhibitory metabolites or residues.

[0033] As used herein, the term "dosage unit form" refers to the physical discrete unit of the agent suitable for the patient to be treated. However, it should be understood that the total daily dosage of the compounds and compositions disclosed herein will be determined by the attending physician within the bounds of reasonable medical judgment. The specific effective dosage level for any particular patient or organism will depend on a variety of factors, including the condition being treated and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health condition, sex, and diet; the timing, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical field.

[0034] compound

[0035] This disclosure includes compounds of formula (I) or formula (II).

[0036]

[0037] or its pharmaceutically acceptable salt

[0038] in

[0039] R 1 It is hydrogen or an optionally substituted C1-C3 alkyl group;

[0040] R 2 It is -SL 2 -R 2a or -OL 2 -R 2a ;

[0041] L2 It is a C1-C3 alkylene group with a bond or optional substitution;

[0042] R 2a Choose from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, and optionally substituted 5-10-membered heteroaryl, wherein R 2a R 2b Replace n examples;

[0043] Each R 2b Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or

[0044] R 2b Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups;

[0045] R 3 Choose from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 5-10-membered heteroaryl, wherein R 3 R 3a Replace m examples;

[0046] Each R 3a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHRc -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or

[0047] R 3a Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups;

[0048] R 4 Choose from the following groups: hydrogen, halogen, -CN, -OH, -OR c -(CH2) 1-3 R c Optionally substituted C2-C6 alkyl groups, optionally substituted C3-C6 carbocyclic groups, optionally substituted 3-10-membered heterocyclic groups, optionally substituted phenyl groups, and optionally substituted 5-10-membered heteroaryl groups, wherein R 4 R 4a Replace p examples;

[0049] Each R 4a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or

[0050] R 4a Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups;

[0051] Each R cIndependently selected from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or

[0052] R c Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups;

[0053] n is 0, 1, 2, 3, 4, or 5; and

[0054] m is 0, 1, 2, 3, 4, or 5; and

[0055] p is 0, 1, 2, 3, 4 or 5.

[0056] This disclosure includes compounds of formula (I) or formula (II).

[0057]

[0058] or its pharmaceutically acceptable salt

[0059] in

[0060] R 1 It is hydrogen or an optionally substituted C1-C3 alkyl group;

[0061] R 2 It is -SL 2 -R 2a or -OL 2 -R 2a ;

[0062] L 2 It is a C1-C3 alkylene group with a bond or optional substitution;

[0063] R 2a Choose from the group consisting of: C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7 membered carbocyclic and optionally substituted 3-10 membered heterocyclic, wherein R 2a R 2b Replace n examples;

[0064] Each R 2b Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c)2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl groups, optionally substituted C2-C6 alkenyl groups, optionally substituted C2-C6 ynyl groups, optionally substituted 3-7 membered carbocyclic groups, and optionally substituted 3-10 membered heterocyclic groups, or

[0065] R 2b Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups;

[0066] R 3 Choose from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 5-10-membered heteroaryl, wherein R 3 R 3a Replace m examples;

[0067] Each R 3a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or

[0068] R 3a Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups;

[0069] R 4 Choose from the following groups: hydrogen, halogen, -CN, -OH, -OR c -(CH2) 1-3 R cOptionally substituted C1-C6 alkyl groups, optionally substituted C3-C6 carbocyclic groups, optionally substituted 3-10-membered heterocyclic groups, optionally substituted phenyl groups, and optionally substituted 5-10-membered heteroaryl groups, wherein R 4 R 4a Replace p examples;

[0070] Each R 4a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or

[0071] R 4a Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups;

[0072] Each R c Independently selected from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or

[0073] R c Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups;

[0074] n is 0, 1, 2, 3, 4, or 5; and

[0075] m is 0, 1, 2, 3, 4, or 5; and

[0076] p is 0, 1, 2, 3, 4 or 5.

[0077] In some embodiments, the compound has formula (Ia) or formula (II-a).

[0078]

[0079] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3a R 4 And m are defined in this paper.

[0080] In some embodiments, the compound has formula (Ib) or formula (II-b).

[0081]

[0082] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3a R 4 And m are defined in this paper.

[0083] In some embodiments, the compound has formula (Ia) or formula (II-c).

[0084]

[0085] Or a pharmaceutically acceptable salt thereof, wherein R 1 R 2 R 3a R 4 And m are defined in this paper.

[0086] R1

[0087] In some implementation schemes, R 1 It is hydrogen or optionally substituted C1-C3 alkyl. In some embodiments, R 1 It is hydrogen. In some implementations, R 1 It is an optionally substituted C1-C3 alkyl group. In some embodiments, R 1 It is hydrogen or optionally substituted methyl. In some embodiments, R 1 It is an optionally substituted methyl group. In some embodiments, R 1 It is methyl. In some embodiments, R 1 It is an optionally substituted ethyl group. In some embodiments, R 1 It is an optionally substituted n-propyl group. In some embodiments, R 1 It is an optional substituted isopropyl group.

[0088] R2

[0089] In some implementation schemes, R 2 It is -SL 2 -R 2a or -OL 2 -R 2a In some implementations, R2 It is -SL 2 -R 2a In some implementations, R 2 Is -OL 2 -R 2a .

[0090] In some implementation schemes, R 2 Choose from the following groups:

[0091] and .

[0092] L2

[0093] In some implementations, L 2 It is a C1-C3 alkylene group, either bonded or optionally substituted. In some embodiments, L 2 It is a key. In some implementations, L 2 It is an optionally substituted C1-C3 alkylene group. In some embodiments, L 2 It is a C1 alkylene group that is either bonded or optionally substituted. In some embodiments, L 2 It is an optionally substituted C1 alkylene group. In some embodiments, L 2 It is an optionally substituted C2-alkylene group. In some embodiments, L 2 It is an optionally substituted C3 alkylene group. In some embodiments, L 2 It is -CH2-.

[0094] R2a

[0095] In some implementation schemes, R 2a Choose from the group consisting of: C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7 membered carbocyclic and optionally substituted 3-10 membered heterocyclic, wherein R 2a R 2b n examples are replaced; in some implementations, R 2a Choose from the group consisting of: optionally substituted C1-C3 alkyl, optionally substituted C2-C3 alkenyl, optionally substituted C2-C4 ynyl, optionally substituted 3-6 membered carbocyclic, optionally substituted phenyl, and optionally substituted 5-6 membered heteroaryl, wherein R 2a R 2b Replace n examples. In some implementations, R 2a It is an optionally substituted C1-C3 alkyl group, wherein R 2a R 2b Replace n examples. In some implementations, R a The group consisting of: optionally substituted methyl, optionally substituted ethyl, and optionally n-propyl. In some embodiments, R 2a It is an optional substituted C2-C3 alkenyl group, wherein R 2a R 2b Replace n examples. In some implementations, R 2a It is an optionally substituted allyl group. In some embodiments, R 2a It is an optional substituted C2-C4 ynyl group, wherein R 2a R 2b Replace n examples. In some implementations, R 2a It is an optionally substituted propyne group. In some embodiments, R 2aIt is an optional substituted 3-6 membered carbon cyclogroup, wherein R 2a R 2b Replace n examples. In some implementations, R 2a It is an optionally substituted cyclopropyl group or an optionally substituted cyclopropyl group.

[0096] R2b

[0097] In some implementations, each R 2b Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7 membered carbocyclic and optionally substituted 3-10 membered heterocyclic, or R 2b Both examples optionally form substituted 4-7 membered heterocyclic groups together with their intercalary atoms. In some embodiments, each R 2b Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbon cycloyl and optionally substituted 3-10-membered heterocyclic groups.

[0098] In some implementations, each R 2b Independently selected from the group consisting of: halogens, -OH groups, and optionally substituted C1-C6 alkyl groups. In some embodiments, each R 2bIndependently selected from the group consisting of: halogens, -OH groups, and optionally substituted C1-C3 alkyl groups. In some embodiments, each R 2b The group consisting of fluorinated groups, chlorolated groups, -OH groups, and optionally substituted methyl groups is selected independently.

[0099] R3

[0100] In some implementation schemes, R 3 Choose from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 5-10-membered heteroaryl, wherein R 3 R 3a The m examples are replaced. In some implementations, R 3 It is an optionally substituted phenyl or an optionally substituted 6-membered heteroaryl group. In some embodiments, R 3 It is an optionally substituted phenyl or optionally substituted pyridyl group. In some embodiments, R 3 It is an optionally substituted 3-7 membered carbon cyclogroup. In some embodiments, R 3 It is an optionally substituted 3-7 membered heterocyclic group. In some embodiments, R 3 It is an optional substituted phenyl group.

[0101] R3a

[0102] In some implementations, each R 3a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or R 3a Both examples optionally form substituted 4-7 membered heterocyclic groups together with their intercalary atoms. In some embodiments, each R 3a Choose independently from the following groups: halogen, -OH, -OR c, -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, and optionally substituted 5-10-membered heteroaryl. In some embodiments, R 3a It is a halogen. In some implementations, R 3a It is a chlorinated or fluorinated group. In some embodiments, R 3a Yes -OMe. In some implementations, R 3a It is an optionally substituted C1-C6 alkyl group. In some embodiments, R 3a It is an optionally substituted methyl group. In some embodiments, R 3a It is -(CH2) 1-3 R c In some implementations, R 3a Yes-S(O)2R c In some implementations, R 3a It is -C(O)N(R) c 2. In some implementations, where R 3a Yes - CN.

[0103] R4

[0104] In some implementation schemes, R 4 Choose from the following groups: hydrogen, halogen, -CN, -OH, -OR c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl groups, optionally substituted C3-C6 carbocyclic groups, optionally substituted 3-10-membered heterocyclic groups, optionally substituted phenyl groups, and optionally substituted 5-10-membered heteroaryl groups, wherein R 4 R 4a The p examples are replaced. In some implementations, R 4 Choose from the following groups: hydrogen, halogen, -CN, -OH, -OR c -(CH2) 1-3 R cOptionally substituted C2-C6 alkyl groups, optionally substituted C3-C6 carbocyclic groups, optionally substituted 3-10-membered heterocyclic groups, optionally substituted phenyl groups, and optionally substituted 5-10-membered heteroaryl groups, wherein R 4 R 4a Replace p examples.

[0105] In some implementation schemes, R 4 It is an optionally substituted C1-C4 alkyl group. In some embodiments, R 4 It is an optionally substituted methyl group. In some embodiments, R 4 It is an optionally substituted C2-C4 alkyl group. In some embodiments, R 4 It is optionally substituted ethyl, optionally substituted isopropyl, optionally substituted tert-butyl, and optionally substituted sec-butyl.

[0106] In some implementation schemes, R 4 It is an optionally substituted 3-7 membered heterocyclic group. In some embodiments, R 4 It is an optionally substituted 5-7 membered heterocyclic group. In some embodiments, R 4 Choose from the group consisting of: optionally substituted tetrahydrofuranyl, optionally substituted tetrahydropyranyl, optionally substituted piperidinyl, optionally substituted morpholinyl and optionally substituted oxazine-heptyl.

[0107] In some implementation schemes, R 4 It is an optionally substituted phenyl group. In some embodiments, R 4 It is an optionally substituted 5-6 membered heteroaryl group. In some embodiments, R 4 The group consisting of: optionally substituted imidazole, optionally substituted pyridyl, optionally substituted pyrimidinyl, and optionally substituted pyrazinyl. In some embodiments, R 4 Choose from the following groups:

[0108] and .

[0109] In some implementation schemes, R 4 Choose from the following groups: hydrogen, methyl, and .

[0110] R4a

[0111] In some implementations, each R 4a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O) OH, -C(O) OR c -C(O) R c -NHR c -N(R) c )2、-C(O) NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or R 4a Two examples, together with their intermediate atoms, optionally form an optionally substituted 4-7 membered heterocyclic group. Each R 4a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R cOptionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, and optionally substituted 5-10-membered heteroaryl. In some embodiments, R 4a Choose from the group consisting of: halogens, optionally substituted C1-C6 alkyl groups, optionally substituted 3-7 membered carbon cycloyl groups, and -C(O)OR. c In some implementations, R 4a Choose from the group consisting of: halogens, optionally substituted C1-C3 alkyl groups, optionally substituted 3-4 membered carbon cycloyl groups, and -C(O)OR. c In some implementations, R 4a Choose from the group consisting of: fluorinated groups, optionally substituted methyl groups, optionally substituted ethyl groups, optionally substituted isopropyl groups, optionally substituted cyclopropyl groups, and -Boc groups.

[0112] This disclosure includes the compounds in Table 1 or their pharmaceutically acceptable salts.

[0113] Table 1

[0114]

[0115]

[0116]

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155] Alternative implementation plan

[0156] In alternative embodiments, the compounds described herein may also contain one or more isotopic substitutions. For example, hydrogen may be... 2 H (D or deuterium) or 3 H (T or tritium); carbon can be, for example... 13 C or 14 C; oxygen can be, for example 18 O; nitrogen can be, for example 15 N, etc. In other embodiments, specific isotopes (e.g., 3 H, 13 C 14 C 18 O or 15 N) can represent at least 1%, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 99.9% of the total isotopic abundance of the element occupying a specific site in the compound.

[0157] Unless otherwise stated, the stereochemistry of the compounds and exemplary synthetic intermediates in Table 1 is arbitrarily specified for clarity. Furthermore, this disclosure contemplates tautomers, rotational isomers, and conformational isomers of the compounds shown herein.

[0158] This disclosure includes racemic mixtures of any compounds disclosed herein.

[0159] Pharmaceutical Composition

[0160] In some embodiments, this disclosure provides compositions comprising compounds of formulas (I) and (II) and pharmaceutically acceptable carriers, adjuvants, or mediators. In some embodiments, the amount of the compound in the compositions contemplated herein enables effective and measurable treatment of a disease or condition in a biological sample or patient. In some embodiments, the amount of the compound in the compositions of this disclosure enables effective and measurable treatment of a disease or condition in a biological sample or patient. In some embodiments, the compositions contemplated herein are formulated for administration to a patient in need of such compositions. In some embodiments, the compositions contemplated herein are formulated for oral administration to a patient.

[0161] In some embodiments, the compositions of this disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implantable reservoir. In some preferred embodiments, the compositions may be administered orally, intraperitoneally, or intravenously. In some embodiments, the sterile injectable form of a composition comprising one or more compounds of formula (I) and formula (II) may be an aqueous or oily suspension. In some embodiments, the suspension may be formulated using suitable dispersants or wetting agents and suspending agents according to techniques known in the art. In some embodiments, the sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable diluent or solvent, such as a solution of 1,3-butanediol. In some embodiments, acceptable media and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. In some embodiments, additional examples include, but are not limited to, sterile, non-volatile oils conventionally used as solvents or suspension media.

[0162] As used in this article, the term "extragastric" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrasheath, intrahepatic, intralesional, and intracranial injection or infusion techniques.

[0163] Pharmaceutically acceptable compositions comprising one or more compounds of formula (I) and formula (II) may be administered orally in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions, or solutions. In some embodiments, the carriers used include lactose and corn starch. Lubricants such as magnesium stearate are also commonly added. In some embodiments, useful diluents include lactose and dried corn starch. In some embodiments, when an aqueous suspension is required for oral administration, the active ingredient is combined with an emulsifier and a suspending agent. In some embodiments, certain sweeteners, flavoring agents, or coloring agents may also be added.

[0164] Alternatively, pharmaceutically acceptable compositions comprising compounds of formulas (I) and (II) can be used for administration in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and thus melts in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol.

[0165] Pharmaceutically acceptable compositions comprising compounds of formulas (I) and (II) may also be applied topically, particularly when the therapeutic target includes areas or organs easily accessible by topical application, including diseases of the eyes, skin, or lower intestine. Suitable topical formulations are readily prepared for each of these areas or organs. In some embodiments, the pharmaceutically acceptable compositions may be formulated in suitable ointments containing an active ingredient suspended or dissolved in one or more carriers. Carriers for the topical application of the compounds of this disclosure include, but are not limited to, mineral oils, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsified waxes, and water. Alternatively, the provided pharmaceutically acceptable compositions may be formulated in suitable lotions or creams containing an active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oils, dehydrated sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecyl alcohol, benzyl alcohol, and water.

[0166] Pharmaceutically acceptable compositions comprising compounds of formulas (I) and (II) may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the field of pharmaceutical formulation and may be prepared as saline solutions using benzyl alcohol or other suitable preservatives, bioavailability enhancers, fluorocarbons, and / or other conventional solubilizers or dispersants.

[0167] In some embodiments, the amount of the disclosed compounds that can be combined with a carrier material to produce a single dosage form composition will vary depending on the host being treated and the specific administration method. Preferably, the provided compositions should be formulated such that an inhibitor can be administered to a patient receiving these compositions at a dose between 0.01 and 100 mg / kg body weight / day.

[0168] The currently disclosed compounds can be formulated into pharmaceutical compositions with pharmaceutically acceptable carriers or excipients. In this respect, a pharmaceutical composition is provided comprising a compound of formula (I) or (II) and a pharmaceutically acceptable excipient, diluent, or carrier.

[0169] Formulations of compounds of formula (I) or (II) include those suitable for the routes of administration detailed herein. They can be readily provided in unit doses and can be formulated into pharmaceutical compositions according to standard pharmaceutical practices. General techniques and formulations applicable to this document can be found in Remington's Pharmaceutical Sciences (16th edition, Osol, A. editor (1980); Mack Publishing Co., Easton, PA). Such methods involve the step of combining the active ingredient with an excipient or carrier constituting one or more auxiliary components. Typically, formulations are prepared by homogenizing and tightly binding the active ingredient with a liquid excipient or carrier, or a finely chopped solid excipient or carrier, or both, and then shaping the product as needed.

[0170] Typical formulations are prepared by mixing compounds of formula (I) or (II) with a carrier, diluent, or excipient. Suitable carriers, diluents, and excipients are well known to those skilled in the art and include materials such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, and water. The specific carrier, diluent, or excipient used will depend on the intended use and purpose of the compound of formula (I) or (II). The choice of solvent is generally based on solvents that are considered by those skilled in the art to be safe (GRAS) for administration to mammals. Typically, safe solvents are non-toxic aqueous solvents, such as water and other non-toxic solvents that are soluble or miscible with water. Suitable aqueous solvents include water, ethanol, propylene glycol, polyethylene glycol (e.g., PEG 400, PEG 300), and mixtures thereof. The formulation may also include one or more of the following: buffers, stabilizers, surfactants, humectants, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, flow aids, processing aids, colorants, sweeteners, flavorings, diluents, and other known additives to provide an appealing appearance for the drug (i.e., a compound of formula (I) or (II) or a pharmaceutical composition thereof) or to aid in the manufacture of a pharmaceutical product (i.e., a pharmaceutical preparation).

[0171] Formulations can be prepared using conventional dissolution and mixing procedures. For example, the active pharmaceutical ingredient (i.e., a compound of formula (I) or (II), or a stable form of a compound of formula (I) or (II) (e.g., a complex with a cyclodextrin derivative or other known complexing agent) is dissolved in a suitable solvent in the presence of one or more of the aforementioned excipients. Compounds of formula (I) or (II) are typically formulated into pharmaceutical dosage forms to provide easily controlled drug dosages and enable patients to adhere to prescribed treatment regimens.

[0172] Pharmaceutical compositions (or formulations) for application can be packaged in various ways depending on the method of administration. Typically, articles for dispensing include containers containing the pharmaceutical formulation in a suitable form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), pouches, ampoules, plastic bags, metal cylinders, etc. Containers may also include tamper-evident components to prevent accidental access to the contents. Furthermore, the container contains a label describing its contents. The label may also include appropriate warnings.

[0173] Pharmaceutical formulations for various routes of administration and types can be prepared. For example, compounds of formula (I) or formula (II) with the desired purity may optionally be mixed with pharmaceutically acceptable diluents, carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences (1980) 16th edition, Osol, A. ed.), in the form of lyophilized formulations, ground powders or aqueous solutions.

[0174] Formulations can be made by mixing with physiologically acceptable excipients or carriers (i.e., excipients or carriers that are non-toxic to the recipient at the dose and concentration used) at ambient temperature, appropriate pH, and desired purity. The pH of the formulation depends primarily on the specific application and the concentration of the compound, but can range from about 3 to about 8. Formulations in acetate buffer at pH 5 are a suitable implementation.

[0175] Compounds of formula (I) or (II) may be sterile. Specifically, formulations intended for in vivo administration should be sterile. This sterility can be readily achieved through filtration via a sterile filter membrane.

[0176] Compounds of formula (I) or (II) can typically be stored as solid compositions, lyophilized formulations, or aqueous solutions.

[0177] Pharmaceutical compositions comprising compounds of formula (I) or (II) may be formulated, administered, and applied in accordance with good medical practice (i.e., dosage, concentration, schedule, duration, medium, and route of administration). Factors to be considered in this context include the specific condition being treated, the specific mammal being treated, the individual patient's clinical condition, the cause of the condition, the site of delivery, the method of administration, the timing of administration, and other factors known to the medical practitioner. The "therapeuticly effective amount" of the compound to be administered will be governed by these considerations and is the minimum amount necessary to prevent, improve, or treat clotting factor-mediated conditions. In some embodiments, this amount is below levels that would be toxic to the host or make the host more susceptible to bleeding.

[0178] Acceptable diluents, carriers, excipients, and stabilizers are non-toxic to the recipient at the doses and concentrations used and include buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (such as octadecyl dimethyl benzyl ammonium chloride; hexamethyl diammonium chloride; benzalkonium chloride; benzyl chloride; phenol, butyl or benzyl alcohol; alkyl esters of p-hydroxybenzoate such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight peptides (less than about 10 residues); proteins. The active pharmaceutical ingredient may be encapsulated in microcapsules, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, such as sodium ions; metal complexes (such as Zn-protein complexes); and / or nonionic surfactants, such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG). The active pharmaceutical ingredient may also be encapsulated in microcapsules (e.g., hydroxymethyl cellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively) prepared by cohesive techniques or interfacial polymerization, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or in crude emulsions. This technology was disclosed in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. (1980).

[0179] Sustained-release preparations of compounds of formula (I) or (II) can be prepared. Suitable examples of sustained-release preparations include semi-permeable matrices of solid hydrophobic polymers containing compounds of formula (I) or (II), said matrices being in the form of molded articles, such as membranes or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactides, copolymers of L-glutamic acid and γ-ethyl-L-glutamic acid, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers (such as LUPRON DEPOT™) (injectable microspheres composed of lactic acid-glycolic acid copolymers and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid.

[0180] Formulations of compounds of formula (I) or formula (II) suitable for oral administration can be prepared in discrete units, such as pills, capsules, sachets or tablets, each unit containing a predetermined amount of the compound of formula (I) or formula (II).

[0181] Compressed tablets can be prepared by compressing an active ingredient, such as powder or granules, in a free-flowing form, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant, or dispersant, in a suitable machine. Molded tablets can be prepared by molding a mixture of powdered active ingredients moistened with an inert liquid diluent in a suitable machine. Tablets can optionally be coated or scored, and optionally formulated to provide a slow or controlled release of the active ingredient.

[0182] Tablets, lozenges, tablets, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules (e.g., gelatin capsules), syrups, or elixirs can be prepared for oral use. Formulations of compounds of formula (I) or (II) intended for oral use can be prepared according to any method known in the art for manufacturing pharmaceutical compositions, and such compositions may contain one or more agents including sweeteners, flavoring agents, coloring agents, and preservatives to provide a palatable preparation. Tablets containing an active ingredient mixed with pharmaceutically acceptable, non-toxic excipients suitable for use in the manufacture of tablets are acceptable. These excipients may be, for example, inert diluents such as calcium carbonate or sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrants such as corn starch or alginic acid; binders such as starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. Tablets may be uncoated or coated using known techniques, including microencapsulation, to delay disintegration and absorption in the gastrointestinal tract, thereby providing a prolonged effect over a longer period. For example, delaying materials such as glyceryl monostearate alone, glyceryl distearate alone, or glyceryl monostearate alone, or glyceryl distearate with wax, may be used.

[0183] For the treatment of the eyes or other external tissues (e.g., the mouth and skin), the formulation is preferably applied as a topical ointment or cream containing, for example, 0.075 to 20% w / w of the active ingredient. When formulated as an ointment, the active ingredient may be used with a paraffin or water-miscible ointment base. Alternatively, the active ingredient may be formulated as a cream with an oil-in-water emulsion base.

[0184] If desired, the aqueous phase of the cream base may include polyols, i.e., alcohols having two or more hydroxyl groups, such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerin, and polyethylene glycol (including PEG 400) and mixtures thereof. Topical formulations may ideally include compounds that enhance the absorption or penetration of the active ingredient across the skin or other infected areas. Examples of such skin penetration enhancers include dimethyl sulfoxide and related analogues.

[0185] The oil phase of an emulsion may be composed of known ingredients in a known manner. While the phase may contain only an emulsifier, it may contain at least one emulsifier in a mixture of fats or oils, or in a mixture of both. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. One or more emulsifiers, with or without one or more stabilizers, form a so-called emulsified wax, and the wax, together with oils and fats, forms a so-called emulsified ointment matrix, which forms the oily dispersed phase of the cream formulation. Emulsifiers and emulsion stabilizers suitable for formulations include Tween® 60, Span® 80, cetearyl alcohol, benzyl alcohol, myristyl alcohol, glyceryl monostearate, and sodium lauryl sulfate.

[0186] Aqueous suspensions of compounds of formula (I) or (II) contain active materials mixed with excipients suitable for manufacturing aqueous suspensions. Such excipients include suspending agents such as sodium carboxymethyl cellulose, croscarmellose, povidone, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic; and dispersants or wetting agents such as naturally occurring phospholipids (e.g., lecithin), condensation products of alkyl esters and fatty acids (e.g., polyoxyethylene stearate), condensation products of ethylene oxide and long-chain aliphatic alcohols (e.g., heptadecetylidene cetyl alcohol), and condensation products of ethylene oxide and esters derived from fatty acids and hexyl anhydrides (e.g., polyoxyethylene sorbitan monooleate). Aqueous suspensions may also contain one or more preservatives such as ethylparaben or n-propylparaben, one or more colorants, one or more flavoring agents, and one or more sweeteners such as sucrose or saccharin.

[0187] Pharmaceutical compositions of compounds of formula (I) or (II) can be in the form of sterile injectable preparations, such as sterile injectable aqueous or oily suspensions. Such suspensions can be formulated according to known techniques using suitable dispersants or wetting agents and suspending agents already mentioned above. Sterile injectable preparations can also be sterile injectable solutions or suspensions in non-toxic, parenteral-acceptable diluents or solvents (such as 1,3-butanediol). Sterile injectable preparations can also be prepared as lyophilized powders. Acceptable media and solvents include water, Ringer's solution, and isotonic sodium chloride solution. Furthermore, sterile non-volatile oils are routinely used as solvents or suspension media. For this purpose, any mild non-volatile oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids (such as oleic acid) can also be used in the preparation of injectable solutions.

[0188] The amount of active ingredient that can be combined with excipients or carrier materials to produce a single dosage form will vary depending on the host being treated and the specific administration method. For example, a sustained-release formulation intended for oral administration to humans may contain about 1 to 1000 mg of the active material, mixed with an appropriate and convenient amount of excipients or carrier materials, which may comprise about 5% to about 95% (by weight) of the total composition. Pharmaceutical compositions can be prepared to provide easily measurable dosages. For example, an aqueous solution intended for intravenous infusion may contain about 3 to 500 pg of the active ingredient per milliliter of solution to enable the infusion of a suitable volume at a rate of about 30 mL / h.

[0189] Preparations suitable for parenteral administration include aqueous and non-aqueous sterile injectable solutions that may contain antioxidants, buffers, antibacterial agents, and solutes that make the preparation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions that may contain suspending agents and thickeners.

[0190] Formulations suitable for topical application to the eyes also include eye drops, wherein the active ingredient is dissolved or suspended in a suitable excipient or carrier, particularly in an aqueous solvent for the active ingredient. The active ingredient is preferably present in such formulations at a concentration of about 0.5% to 20% w / w, for example, about 0.5% to 10% w / w, for example, about 1.5% w / w.

[0191] Formulations suitable for topical application in the oral cavity include: tablets containing an active ingredient in a flavoring matrix (typically sucrose and gum arabic or tragacanth); soft tablets containing an active ingredient in an inert matrix (such as gelatin and glycerin or sucrose and gum arabic); and mouthwashes containing an active ingredient in a suitable liquid carrier.

[0192] Formulations for rectal administration may be provided in suppository form having a suitable matrix comprising, for example, cocoa butter or salicylates.

[0193] Formulations suitable for intrapulmonary or nasal administration have particle sizes ranging from, for example, 0.1 to 500 micrometers (in increments such as 0.5, 1, 30, 35 micrometers, etc.) and are administered by rapid inhalation through the nasal passage or by oral inhalation to reach the alveolar sacs. Suitable formulations include aqueous or oil solutions of the active ingredient. Formulations suitable for aerosol or dry powder administration can be prepared according to conventional methods and can be delivered together with other therapeutic agents, such as compounds used to date for the treatment or prevention of the conditions described below.

[0194] Preparations suitable for vaginal application may be provided in the form of vaginal suppositories, tampons, creams, gels, pastes, foams or sprays, and in addition to the active ingredient, contain excipients or carriers as appropriate in the art.

[0195] Formulations can be packaged in single-dose or multi-dose sealed containers, such as sealed ampoules and vials, and can be stored under lyophilized (freeze-dried) conditions, requiring only the immediate addition of a sterile liquid excipient or carrier, such as water for injection, before use. Temporary injectable solutions and suspensions can be prepared from sterile powders, granules, and tablets of the types described above. Preferred single-dose formulations are those containing the active ingredient at the daily dose or a daily sub-dose or a suitable portion thereof as described herein.

[0196] The present invention further provides veterinary compositions comprising at least one active ingredient as defined above and a veterinary excipient or carrier. The veterinary excipient or carrier is a material for administering the composition, which may be a solid, liquid, or gaseous material that is inert or acceptable in the veterinary field and compatible with the active ingredient. These veterinary compositions may be administered via parenteral, oral, or any other desired route.

[0197] In specific embodiments, the pharmaceutical composition comprising the compounds disclosed herein further comprises a chemotherapeutic agent. In some of these embodiments, the chemotherapeutic agent is an immunotherapeutic agent.

[0198] Reagent test kit

[0199] A kit is also provided for carrying out the methods detailed herein, the kit comprising one or more of the compounds described herein or a pharmaceutical composition comprising the compounds described herein. The kit may employ any of the compounds disclosed herein. In one variation, the kit employs the compounds described herein or a pharmaceutically acceptable salt thereof. The kit may be used for any one or more of the uses described herein and may therefore contain instructions for use for treating conditions such as cancer. In some embodiments, the kit contains instructions for use for treating cancer.

[0200] Kits typically include suitable packaging. A kit may include one or more containers containing any of the compounds described herein. Each component (if there is more than one component) may be packaged in a separate container, or, where cross-reactivity and shelf life permit, some components may be combined in one container. One or more components of the kit may be sterile and / or may be contained in sterile packaging.

[0201] The kit may be available in unit dose, bulk packaging (e.g., multi-dose packaging), or subunit dose form. For example, a kit may be provided containing a sufficient dose of a compound as disclosed herein (e.g., a therapeutically effective amount) and / or a second pharmaceutically active compound for treating a condition (e.g., cancer) to provide effective treatment to an individual over a longer period (such as any time such as one week, two weeks, three weeks, four weeks, six weeks, eight weeks, three months, four months, five months, seven months, eight months, nine months, or longer). The kit may also include multiple unit doses of the compound and instructions for use, and may be packaged in sufficient quantities for storage and use in pharmacies (e.g., hospital pharmacies and dispensing pharmacies).

[0202] The kit may optionally include a set of instructions, typically written, although electronic storage media (e.g., disks or optical discs) containing instructions relating to the use of the components of the methods disclosed herein are also acceptable. The instructions accompanying the kit typically include information about the components and their administration to the subject.

[0203] Methods using the compounds disclosed herein

[0204] In some embodiments, this disclosure provides a method for treating or reducing the severity of a disease or disorder associated with α-synuclein. In some embodiments, a method is used to treat α-synucleinopathy, reduce the severity of α-synucleinopathy, or prevent α-synucleinopathy. In some embodiments, this disclosure provides a method for treating or reducing the severity of a disease or disorder associated with α-synuclein, the method comprising the step of administering a composition according to this disclosure to the patient. In some embodiments, this disclosure provides a method for treating or reducing the severity of a disease or disorder associated with α-synuclein, the method comprising the steps of selecting a composition according to this disclosure and administering the composition according to this disclosure to the patient.

[0205] As used herein, the term "disease or disorder associated with α-synuclein" means any disease or other harmful disorder in which α-synuclein or α-synuclein misfolding is known to play a role. Therefore, another embodiment of this disclosure relates to treating or reducing the severity of one or more diseases in which α-synuclein or α-synuclein misfolding is known to play a role. Therefore, another embodiment of this disclosure relates to treating or preventing one or more diseases in which α-synuclein or α-synuclein misfolding is known to play a role. In some embodiments, the disease or disorder associated with α-synuclein is a neurodegenerative disease.

[0206] As used herein, the term "Lewy body dementia" refers to any disease or other debilitating disorder in which dementia occurs with the deposition, aggregation, or accumulation of Lewy bodies in the brain. Therefore, another embodiment of this disclosure relates to treating or reducing the severity of one or more diseases in which Lewy body dementia is known to play a role. Therefore, another embodiment of this disclosure relates to preventing or reducing the severity of one or more diseases in which Lewy body dementia or Lewy body dementia protein misfolding plays a role. In some embodiments, the disease or disorder associated with Lewy body dementia is a neurodegenerative disease. In some embodiments, the dementia-related disease or disorder does not occur with Lewy bodies.

[0207] In some embodiments, application of the compounds of this disclosure causes prevention of protein misfolding. In some embodiments, application of the compounds of this disclosure causes prevention of α-synuclein misfolding. In some embodiments, this disclosure causes treatment or prevention of diseases or disorders associated with the formation, deposition, accumulation, or persistence of α-synuclein oligomers, proto-fibrils, fibrils, or aggregates.

[0208] In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 10-100%. In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 20-100%. In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 30-100%. In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 40-100%. In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 50-100%. In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 60-100%. In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 70-100%. In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 80-100%. In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 90-100%. In some embodiments, preventing protein misfolding is defined as reducing protein misfolding by 100%.

[0209] In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 10-100% in protein oligomers, fibrils, fibers, or aggregates. In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 20-100% in protein oligomers, fibrils, fibers, or aggregates. In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 30-100% in protein oligomers, fibrils, fibers, or aggregates. In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 40-100% in protein oligomers, fibrils, fibers, or aggregates. In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 50-100% in protein oligomers, fibrils, fibers, or aggregates. In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 60-100% in protein oligomers, fibrils, fibers, or aggregates. In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 70-100%. In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 80-100%. In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 90-100%. In some embodiments, preventing the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates is defined as a reduction of 100%.

[0210] In some embodiments, the compounds and compositions according to the methods of this disclosure may be administered in any amount and via any route of administration to effectively treat protein misfolding or alleviate its severity. The exact amount required will vary depending on the subject, which may be determined by the subject's species, age and general condition, the severity of the infection, the specific agent, the mode of administration, etc. For ease of administration and uniformity of dosage, the compounds of this disclosure are preferably formulated in dose-unit form.

[0211] In some embodiments, the compounds and compositions according to the methods of this disclosure may be administered in any amount and via any route of administration to effectively treat or reduce the formation, deposition, accumulation, or persistence of protein oligomers, fibrils, fibers, or aggregates, or to alleviate their severity. The exact amount required will vary depending on the subject, taking into account the subject's species, age and general condition, the severity of the infection, the specific agent, the mode of administration, etc. For ease of administration and uniformity of dosage, the compounds of this disclosure are preferably formulated in unit dosage form.

[0212] In some embodiments, the compounds of this disclosure act as antibodies. In some embodiments, the compounds of this disclosure bind to at least a portion of α-synuclein and prevent the aggregation of at least a portion of α-synuclein. In some embodiments, the compounds of this disclosure act as antibodies. In some embodiments, the compounds of this disclosure bind to at least a portion of α-synuclein and prevent the binding of at least a portion of α-synuclein to a cell receptor. In some embodiments, the compounds of this disclosure bind to at least a portion of misfolded α-synuclein and prevent the aggregation of at least a portion of misfolded α-synuclein. In some embodiments, the compounds of this disclosure bind to at least a portion of misfolded α-synuclein and prevent the binding of at least a portion of misfolded α-synuclein to a cell receptor.

[0213] In some embodiments, the compounds disclosed herein and one or more additional therapeutic agents may be formulated into a single dosage form.

[0214] In some embodiments, the compounds of this disclosure inhibit or reduce the severity of α-synuclein-induced cytotoxicity. In some embodiments, cytotoxicity in human neurons and glial cells is reduced or inhibited. In some embodiments, the compounds of this disclosure inhibit or reduce the severity of intercellular transmission of oligo-α-synuclein. In some embodiments, intercellular transmission between neurons and glial cells is reduced or inhibited. In some embodiments, the compounds of this disclosure inhibit or reduce caspase production. In some embodiments, caspase production in neurons and glial cells is reduced or inhibited.

[0215] In some embodiments, the compounds or pharmaceutical compositions of this disclosure are used to treat one or more diseases or disorders associated with the formation, deposition, accumulation, or persistence of α-synuclein oligomers, fibrils, fibers, or aggregates, said diseases or disorders being selected from the group consisting of: Parkinson's disease, Parkinson's dementia, dementia associated with Lewy bodies, diffuse Lewy body disease, sporadic and familial Alzheimer's disease, Lewy body variants of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher's disease. Diseases, including systemic axonal dystrophy with adolescent onset, Hallervorden-Spatz disease, pure autonomic failure, neurodegeneration associated with type 1 brain iron stasis, generalized neurodegenerative diseases, synucleinosis, pure autonomic failure, dementia, tau proteinosis, arginophilic granuloma, Pick's disease, Down syndrome, chronic leukemia, lymphoma, schizophrenia, psychosis, or Creutzfeldt-Jakob's disease.

[0216] In some implementations, the compound is administered to the subject at doses between about 0.001 pg / kg and about 1,000 mg / kg, including but not limited to about 0.001 pg / kg, about 0.01 pg / kg, about 0.05 pg / kg, about 0.1 pg / kg, about 0.5 pg / kg, about 1 pg / kg, about 10 pg / kg, about 25 pg / kg, about 50 pg / kg, about 100 pg / kg, about 250 pg / kg, about 500 pg / kg, about 1 mg / kg, about 5 mg / kg, about 10 mg / kg, about 25 mg / kg, about 50 mg / kg, about 100 mg / kg, and about 200 mg / kg.

[0217] example

[0218] The compounds disclosed herein can be synthesized by analogy with methods known to those skilled in the art and by analogy with the synthesis of the specific compounds described in the following examples.

[0219] abbreviation

[0220]

[0221] Synthesis Method A

[0222] Example 1 - 7-Methyl-8-phenyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0223]

[0224] Step 1: α-Acetyl-phenylacetonitrile

[0225] NaH (7.43 g, 185 mmol, 60% in mineral oil) was added fractionally to a stirred solution of phenylacetonitrile (11.7 g, 99.9 mmol) in THF (200 mL) at 0 °C under N2. The resulting mixture was stirred at 0 °C under N2 for 15 min. EA (35 g, 400 mmol) was added to the mixture at 0 °C. The resulting mixture was stirred overnight at room temperature. The reaction was quenched with 2N HCl (100 mL), and the resulting mixture was extracted with EA (3 × 100 mL). The combined organic layers were washed with brine (80 mL) and dried over Na2SO4. After filtration, the filtrate was concentrated to give a crude product, which was purified on silica gel (PE / EA = 10 / 1) to give the title compound (14.8 g, 93%) as a grayish-white solid. LCMS (ES, m / z): [MH] - =158.

[0226] Step 2: 5-Methyl-4-phenyl-2H-pyrazole-3-amine

[0227] At room temperature, N₂H₄-H₂O (2.5 g, 40 mmol, 80 wt%) was added to a stirred solution of α-acetyl-phenylacetonitrile (3.18 g, 20.0 mmol) in EtOH (30 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure and diluted with water (50 mL). The mixture was extracted with EA (3 × 70 mL), and the combined organic layers were washed with brine (2 × 40 mL) and dried over Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (3.07 g, 89%), which could be used directly in the next step without further purification. LCMS (ES, m / z): [M+H] + =174.

[0228] Step 3: N-[(5-methyl-4-phenyl-2H-pyrazol-3-yl)carbamoyl]carbamate

[0229] At room temperature, ethyl N-thiocarbonylcarbamate (930 mg, 7.09 mmol) was added to a stirred solution of 5-methyl-4-phenyl-2H-pyrazole-3-amine (3.07 g, 17.7 mmol) in DMF (53 mL). The resulting mixture was stirred for 1 h. The mixture was then poured into ice water (400 mL). The precipitated solid was collected by filtration and washed with water (3 x 20 mL). The solid was dried under vacuum to give the title compound (2.58 g, 48%) as a yellow solid, which could be used in the next step without further purification. LCMS (ES, m / z): [M+H] + =305.

[0230] Step 4: 7-Methyl-8-phenyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0231] At room temperature, EtONa (5.54 mL, 14.1 mmol, 20 wt% in EtOH) was added to a stirred solution of N-[(5-methyl-4-phenyl-2H-pyrazol-3-yl)aminothioyl]carbamate (2.15 g, 7.06 mmol) in EtOH (30 mL). The resulting mixture was refluxed at 80 °C for 0.5 h. The mixture was filtered, and the filter cake was dissolved in water (20 mL) and acidified to pH 5 with 2 N HCl (8 mL). The precipitated solid was collected by filtration and washed with water (2 × 15 mL) to give the title compound (1.66 g, 91%), which could be used directly in the next step without further purification. LCMS (ES, m / z): [M+H]+ = 259

[0232] Step 5: 7-Methyl-8-phenyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (Example 1)

[0233] Proprynne bromide (111 mg, 0.929 mmol) was added to a stirred mixture of 7-methyl-8-phenyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (400 mg, 1.55 mmol) and K₂CO₃ (642 mg, 4.65 mmol) in DMF (5 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The mixture was filtered, and the filter cake was washed with DMF (2 x 2 mL). The filtrate was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 36% B to 59% B, 59% B over 8 min; wavelength: 220 nm; RT1 (min): 6.13), yielding the title compound (287 mg, 62.54%) as a white solid. LCMS (ES, m / z): [M+H] + = 297. 1H NMR (400 MHz, DMSO-d6): δ 12.95 (s, 1H), 7.77 – 7.75 (m, 2H), 7.47 – 7.43 (m, 2H), 7.32 – 7.28 (m, 1H), 4.01 (d, J =2.6 Hz, 2H), 3.28 – 3.27 (t, J = 2.6 Hz, 1H), 2.47 (s, 3H).

[0234] The embodiments in Table A were synthesized using synthesis method A.

[0235] Table A

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250] Example 85 - 2-[(2-fluorophenyl)thioalkyl]-7-methyl-8-phenyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0251]

[0252] At room temperature, Cu(OAc)₂ (70 mg, 0.39 mmol) and 1,10-phenanthroline (140 mg, 0.774 mmol) were added to a mixture of 7-methyl-8-phenyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (100 mg, 0.387 mmol) and 2-fluorophenylboronic acid (163 mg, 1.16 mmol) in anhydrous DCE (2 mL). The final reaction mixture was irradiated with microwaves at 85 °C for 10 min under N₂. The reaction mixture was cooled to RT. The resulting mixture was filtered, and the filter cake was washed with DCE (2 x 2 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. The crude product was purified by Prep-HPLC under the following conditions (column: Xcelect CSH F-pheny OBD column, 19*250 mm, 5 μm; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 38% B to 64% B, 64% B over 8 min; wavelength: 254 nm; RT1 (min): 6.77), yielding the title compound (120 mg, 88%) as a white solid. LCMS (ES, m / z): [M+H] + =353. 1H NMR (400 MHz, DMSO-d6): δ 13.21 (s, 1H), 7.72 – 7.70 (m, 2H), 7.49 (m, 1H), 7.41 – 7.36 (m, 3H), 7.26 – 7.18 (m, 3H), 2.45 (s, 3H).

[0253] Example 86 - 8-(4-fluorophenyl)-2-(pyridin-2-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0254]

[0255] 2-Iodopyridine (31 mg, 0.15 mmol) was added to a stirred mixture of 8-(4-fluorophenyl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (100 mg, 0.381 mmol), K₂CO₃ (158 mg, 1.14 mmol), and CuI (808 mg, 0.42 mmol) in DMF (2 mL). The resulting mixture was stirred at 120 °C for 1 h. The mixture was then cooled to room temperature. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 10 mL), and the combined organic layers were washed with water (2 x 10 mL) and brine (10 mL) and dried over Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire prep C18 OBD column, 19*150 mm, 5 μm; mobile phase A: water (0.05% FA), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 37% B to 53% B, 53% B over 8 min; wavelength: 220 nm; RT1 (min): 7.13), yielding the title compound (37 mg, 29%) as a white solid. LCMS (ES, m / z): [M+H] + =340. 1 H NMR (400 MHz, DMSO-d6): δ 8.70 – 8.68 (m, 1H), 8.51(s, 1H), 8.03 – 8.01 (m, 1H), 7.92 (d, J = 7.9 Hz, 1H), 7.70 – 7.62 (m, 3H),7.11 – 7.07 (m, 2H).

[0256] Example 87-2-[(4-hydroxybut-2-yn-1-yl)thioalkyl]-7-methyl-8-phenyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0257]

[0258] Step 1: 4-Bromobut-2-yn-1-ol

[0259] At 0 °C, p-toluenesulfonyl chloride (1.78 g, 9.35 mmol) was added in a single batch to a solution of butynediol (3.5 g, 41 mmol), triethylamine (1.32 g, 13.0 mmol), and triethylamine hydrochloride (0.11 g, 0.81 mmol) in ACN (30 mL). The reaction mixture was stirred at 0 °C for 40 min, heated to RT, and quenched with water (150 mL) and EA (150 mL). The aqueous layer was extracted with EA (100 mL). The combined organic matter was washed with aqueous NH4Cl (80 mL) and brine (100 mL) and dried over Na2SO4. The solvent was evaporated under vacuum, and the crude material was purified by column chromatography (EA / PE = 3 / 7) to give the intermediate monotoluenesulfonyl diol, a pale yellow oil. It was dissolved in acetone (10 mL), and then LiBr (1.41 g, 16.3 mmol) was added at room temperature. The mixture was stirred for 1 h, then diluted with Et₂O (50 mL) and water (25 mL). The organic layer was washed with brine (5 mL) and dried over Na₂SO₄. The solvent was evaporated under reduced pressure; no mass signal was detected, and the crude substance, i.e., the title compound (1.5 g, 25%), was ready for use in the next step without further purification.

[0260] Step 2: 2-[(4-hydroxybut-2-yn-1-yl)thioalkyl]-7-methyl-8-phenyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0261] At room temperature under N2, K2CO3 (289 mg, 2.09 mmol) was added to a stirred solution of 7-methyl-8-phenyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (180 mg, 0.697 mmol) in DMF (3.5 mL), followed by dropwise addition of 4-bromobut-2-yn-1-ol (52 mg, 0.35 mmol). The resulting mixture was stirred at room temperature under N2 for 1 h. The resulting mixture was filtered, and the filtrate was purified by Prep-HPLC under the following conditions (column: XSelect CSH Prep C18OBD column, 19*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 19% B to 51% B, 51% B over 8 minutes; wavelength: 254 / 220 nm; RT1 (min): 6.88), yielding the title compound (110 mg, 48%) as a white solid. LCMS (ES, m / z): [M+H]+ =327. 1H NMR (400 MHz, DMSO-d6): δ12.95 (s, 1H), 7.75 (d, J = 7.7 Hz, 2H), 7.46 (t, J = 7.3 Hz, 2H), 7.30 (t, J= 7.3 Hz, 1H), 5.19 (s, 1H), 4.09 (s, 2H), 4.06 (d, J = 2.4 Hz, 2H), 2.56 (s, 3H).

[0262] Example 88 - 8-(4-fluorophenyl)-7-isopropyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0263]

[0264] Step 1: 2-(4-fluorophenyl)-4-methyl-3-oxopentanilide

[0265] NaH (16.5 g, 688 mmol) was added dropwise to a stirred solution of 30 g (222 mmol) of 4-fluorophenylacetonitrile in 300 ml of THF at 0 °C. The resulting mixture was stirred at 0 °C for 20 min. Ethyl isobutyrate (103 g, 888 mmol) was added dropwise to the mixture at 0 °C. The resulting mixture was stirred again overnight at room temperature. The mixture was concentrated under reduced pressure. The mixture was acidified to pH 6 with 2N HCl. The mixture was extracted with EA (3 x 100 ml). The combined organic layers were washed with brine (2 x 100 ml) and dried over anhydrous Na2SO4. The filtrate was concentrated under reduced pressure after filtration. The residue was purified by silica gel column chromatography, eluting with PE / EA (7:1), to give the title compound (32 g, 70%) as a yellow oil. LCMS (ES, m / z): [MH] - =204.

[0266] Step 2: 4-(4-fluorophenyl)-5-isopropyl-2H-pyrazole-3-amine

[0267] Hydrazine hydrate (80%) (15.6 g, 312 mmol) was added to a stirred solution of 2-(4-fluorophenyl)-4-methyl-3-oxopentanonitrile (32 g, 156 mmol) in EtOH (320 mL) at room temperature. The resulting mixture was stirred overnight at 80 °C. The mixture was concentrated under reduced pressure. The mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN aqueous solution (0.05% NH₄⁺). 3· H₂O), gradient of 30% to 40% over 10 min; detector, UV 254 nm. This produced the title compound (10 g, 29%) as a yellow solid. LCMS (ES, m / z): [M+H] + =220.

[0268] Step 3: N-{[4-(4-fluorophenyl)-5-isopropyl-2H-pyrazol-3-yl]aminothioyl}carbamate

[0269] At room temperature, ethyl N-thiocarbonylcarbamate (15.3 g, 116 mmol) was added to a stirred solution of 4-(4-fluorophenyl)-5-isopropyl-2H-pyrazole-3-amine (17 g, 78 mmol) in DMF (170 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was then poured into water / ice (200 mL). The precipitated solid was collected by filtration and washed with water (4 x 20 mL). This yielded the title compound (24.5 g, 87%) as a yellow solid. LCMS (ES, m / z): [M+H] + =352.

[0270] Step 4: 8-(4-fluorophenyl)-7-isopropyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0271] EtONa (9.52 g, 140 mmol) was added to a stirred solution of N-{[4-(4-fluorophenyl)-5-isopropyl-2H-pyrazol-3-yl]aminothioyl}carbamate (24.5 g, 70 mmol) in EtOH (245 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure. The mixture was acidified to pH 6 with 2N HCl. The precipitated solid was collected by filtration and washed with water. This yielded the title compound (22.5 g, 99%) as a yellow solid. LCMS (ES, m / z): [M+H] + =305.

[0272] Step 5: 8-(4-fluorophenyl)-7-isopropyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0273] At room temperature, K₂CO₃ (22.9 g, 166 mmol) and propargyl bromide (7.39 g, 62.1 mmol) were added fractionally to a stirred solution of 8-(4-fluorophenyl)-7-isopropyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (25.2 g, 82.8 mmol) in DMF (120 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was cooled to 0 °C and poured into water at 0 °C. The precipitated solid was collected by filtration and washed with water (3 x 30 mL). The resulting solid was dried under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (12:1) to give a yellow solid (20 g crude). 15 g of the residue was purified by grinding with EA (24 mL) at 65 °C. The mixture was cooled to 0 °C. The precipitated solid was collected by filtration, washed with EA (4 x 5 mL), and dried under reduced pressure. This yielded the title compound (9.6 g, 34%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =343. 1H NMR (400 MHz, DMSO-d6) δ 12.95 (s, 1H), δ 7.70 – 7.61 (m, 2H), 7.28 (td, J = 8.9, 2.2 Hz, 2H), 3.97 (d, J = 2.6 Hz, 2H), 3.31 (septet, J = 6.8 Hz, 1H), 3.22 (s, 1H), 1.25 (d, J = 6.8 Hz, 6H).

[0274] Example 89-8-cyclohexyl-7-methyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0275]

[0276] Step 1: 2-Cyclohexyl-3-oxobutyronitrile

[0277] At 0 °C and N2, NaH (1.49 g, 62 mmol) and EA (7.0 g, 80 mmol) were added fractionally to a stirred solution of 2-cyclohexylacetonitrile (2.46 g, 20 mmol) in THF (29 mL). The resulting mixture was stirred overnight at 60 °C. The reaction was acidified to pH 6, and the resulting mixture was extracted with EA (3 × 80 mL), washed with brine (50 mL), dried over Na2SO4, and filtered to give the title compound (3.66 g, 99%), which could be used in the next step without further purification. No mass signal was detected.

[0278] Step 2: 4-Cyclohexyl-5-methyl-2H-pyrazole-3-amine

[0279] Hydrazine hydrate (80%, 2.0 mL, 41 mmol) was added fractionally to a stirred solution of 2-cyclohexyl-3-oxobutyronitrile (20 mmol) in EtOH (60 mL) at room temperature. The resulting mixture was stirred overnight at 80 °C under N2. The reaction mixture was cooled to RT, concentrated under vacuum, diluted with water (80 mL), and extracted with EA (3 × 60 mL). The combined organic layers were washed with brine (40 mL), dried over anhydrous Na2SO4, and concentrated to give the title compound (2.0 g, 56%), which was ready for use in the next step without further purification. LCMS (ES, m / z): [M+H]+ = 180.

[0280] Step 3: N-[(4-cyclohexyl-5-methyl-2H-pyrazol-3-yl)carbamate]ethyl carbamate

[0281] N-Thiocarbonylcarbamate (0.59 g, 4.5 mmol) was added dropwise to a stirred solution of 4-cyclohexyl-5-methyl-2H-pyrazole-3-amine (2.0 g, 4.5 mmol) in dimethylformamide (40 mL) at room temperature under N2. The resulting mixture was stirred for 1 h. The mixture was diluted with cold water (400 mL), filtered, and the result was the title compound (3.2 g, 92%), which was ready for use in the next step without further purification. LCMS (ES, m / z): [M+H]+ = 311.

[0282] Step 4: 8-Cyclohexyl-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0283] At room temperature, EtONa (20% solution in EtOH) (6.8 mL, 20 mmol) was added to a solution of N-[(4-cyclohexyl-5-methyl-2H-pyrazol-3-yl)aminothioyl]carbamate (3.2 g, 10 mmol) in EtOH (40 mL). The resulting mixture was stirred at 80 °C for 1 h. The precipitated solid was collected by filtration. The residue was dissolved in water (80 mL). The residue was acidified to pH 6 with 1 M HCl. The precipitated solid was collected by filtration and washed with water (2 × 80 mL), leaving the title compound (2.0 g, 73%), which could be used directly for the next step without further purification. LCMS (ES, m / z): [M+H]+ = 265.

[0284] Step 5: 8-Cyclohexyl-7-methyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0285] At room temperature under N2, K2CO3 (400 mg, 2.89 mmol) was added in portions to a stirred solution of 8-cyclohexyl-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (1.0 g, 3.8 mmol) in DMF (5 mL), followed by dropwise addition of propargyl bromide (180 mg, 1.51 mmol). The resulting mixture was stirred at room temperature under N2 for 1 h. The resulting mixture was filtered, and the filtrate was purified by Prep-HPLC under the following conditions (column: SunFire Prep C18 OBD column, 19*150mm, 5μm; mobile phase A: water (0.1%FA), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 47% B to 52% B, 52% B over 12 min; wavelength: 220 nm; RT1 (min): 9.80), yielding the title compound (31.9 mg, 3%) as a grayish-white solid. LCMS (ES, m / z): [M+H]+ =303. 1H NMR (400 MHz, DMSO-d6): δ 12.66 (s, 1H), 3.99 (d, J = 2.6 Hz, 2H), 3.19 (t, J = 2.6 Hz, 1H), 2.60 – 2.51 (m, 1H), 2.26 (s,3H), 1.92-1.64 (m, 7H), 1.39-1.20 (m, 3H).

[0286] Synthesis Method B

[0287] Example 90 - 8-(4-fluorophenyl)-7-methoxy-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0288]

[0289] Step 1: Ethyl 2-cyano-2-(4-fluorophenyl)acetate

[0290] Sodium ethoxide (1.01 g, 14.8 mmol) was added to a stirred solution of 4-fluorophenylacetonitrile (1.0 g, 7.4 mmol) in diethyl carbonate (10 mL, 85 mmol) at room temperature. The resulting mixture was stirred overnight at 90 °C. The mixture was acidified to pH 6 with 2N HCl. The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (8:1), to give the title compound (1.3 g, 85%) as a yellow oil. LCMS (ES, m / z): [MH] - =206.

[0291] Step 2: 5-Amino-4-(4-fluorophenyl)-1,2-dihydropyrazole-3-one

[0292] At room temperature, hydrazine hydrate (80%) (3.74 g, 74.7 mmol) was added to a stirred solution of ethyl 2-cyano-2-(4-fluorophenyl)acetate (7.74 g, 37.4 mmol) in EtOH (80 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure. The mixture was extracted with EA (3 x 70 mL). The combined organic layers were washed with brine (2 x 70 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (4.8 g, 67%) as a white solid. LCMS (ES, m / z): [M+H] + =194.

[0293] Step 3: tert-butyl 5-amino-4-(4-fluorophenyl)-3-oxo-2H-pyrazole-1-carboxylate

[0294] At room temperature, (Boc)₂O (6.37 g, 29.2 mmol) and Na₂CO₃ (3.1 g, 29 mmol) were added fractionally to a stirred solution of 5-amino-4-(4-fluorophenyl)-1,2-dihydropyrazole-3-one (4.7 g, 24 mmol) in MeOH (47 mL). The resulting mixture was stirred at room temperature for 3 h. The mixture was concentrated under reduced pressure. The residue was dissolved in water. The resulting mixture was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1), to give the title compound (5.98 g, 84%) as a white solid. LCMS (ES, m / z): [M+H] + =294.

[0295] Step 4: tert-butyl 5-amino-4-(4-fluorophenyl)-3-methoxypyrazole-1-carboxylate

[0296] MeI (266 mg, 1.88 mmol) was added to a stirred solution of 5-amino-4-(4-fluorophenyl)-3-oxo-2H-pyrazole-1-carboxylic acid tert-butyl ester (500 mg, 1.71 mmol) and K₂CO₃ (707 mg, 5.12 mmol) in DMF (10 mL) at room temperature. The resulting mixture was stirred overnight at room temperature. MeI (484 mg, 3.41 mmol) was added to the mixture at room temperature. The resulting mixture was stirred for another 6 h at room temperature. The residue was dissolved in water. The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with water (2 x 10 mL) and brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (30 mg, 5.7%) and the title compound (292 mg, 56%), both as off-white solids. LCMS (ES, m / z): [M+H] + =308.

[0297] Step 5: 4-(4-fluorophenyl)-5-methoxy-2H-pyrazole-3-amine

[0298] TFA (1.5 mL) was added to a stirred solution of 360 mg (1.17 mmol) of 5-amino-4-(4-fluorophenyl)-3-methoxypyrazole-1-carboxylic acid tert-butyl ester in DCM (8 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (144 mg, 59%) as a yellow solid. LCMS (ES, m / z): [M+H] + =208.

[0299] Step 6: N-{[4-(4-fluorophenyl)-5-methoxy-2H-pyrazol-3-yl]aminothioyl}carbamate

[0300] At room temperature, ethyl N-thiocarbonylcarbamate (50.6 mg, 0.386 mmol) was added to a stirred solution of 100 mg (0.483 mmol) of 4-(4-fluorophenyl)-5-methoxy-2H-pyrazole-3-amine in 2 mL of DMF. The resulting mixture was stirred at room temperature for 1 h. The reaction mixture was then poured into water at 0 °C. The resulting mixture was filtered, and the filter cake was washed with water. The solid was dried under vacuum. This yielded the title compound (200 mg, 85%) as a yellow solid. LCMS (ES, m / z): [M+H] + =339.

[0301] Step 7: 8-(4-fluorophenyl)-7-methoxy-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0302] EtONa (80.5 mg, 1.18 mmol) was added to a stirred solution of N-{[4-(4-fluorophenyl)-5-methoxy-2H-pyrazol-3-yl]aminothioyl}carbamate (200 mg, 0.591 mmol) in EtOH (4 mL). The resulting mixture was stirred at 80 °C for 1 h. The mixture was concentrated under reduced pressure. The mixture was acidified to pH 6 with 2N HCl. The resulting mixture was filtered, and the filter cake was washed with water. The solid was dried under vacuum. This yielded the title compound (196 mg, 96%) as a yellow solid. LCMS (ES, m / z): [M+H] + =293.

[0303] Step 8: 8-(4-fluorophenyl)-7-methoxy-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0304] Proprynne bromide (64.3 mg, 0.541 mmol) was added to a stirred mixture of 8-(4-fluorophenyl)-7-methoxy-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (198 mg, 0.676 mmol) and K₂CO₃ (280 mg, 2.03 mmol) in DMF (4 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 30% to 80% gradient over 10 min; detector, UV 254 nm. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 35% B to 65% B, 65% B over 8 min; wavelength: 220 nm; RT1 (min): 9.23), yielding the title compound (55 mg, 25%) as a white solid. LCMS (ES, m / z): [M+H] + =331. 1H NMR (300 MHz, DMSO-d6) δ 12.98 (s, 1H), 8.16 – 8.04 (m, 2H), 7.29 – 7.15 (m, 2H), 4.08-4.04 (m, 5H), 3.27 (t, J = 2.6 Hz, 1H).

[0305] The examples in Table B were synthesized using synthesis method B:

[0306] Table B

[0307]

[0308] Example 93-8-(4-fluorophenyl)-6-methyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4,7-dione

[0309]

[0310] Step 1: tert-butyl 5-amino-4-(4-fluorophenyl)-2-methyl-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylate

[0311] MeI (266 mg, 1.88 mmol) was added to a stirred solution of 5-amino-4-(4-fluorophenyl)-3-oxo-2H-pyrazole-1-carboxylic acid tert-butyl ester (500 mg, 1.71 mmol) and K₂CO₃ (707 mg, 5.12 mmol) in DMF (10 mL) at room temperature. The resulting mixture was stirred overnight at room temperature. MeI (484 mg, 3.41 mmol) was added to the above mixture at room temperature. The resulting mixture was stirred for another 6 h at room temperature. The residue was dissolved in water. The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (292 mg, 56%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =308.

[0312] Step 2: 5-Amino-4-(4-fluorophenyl)-2-methyl-1,2-dihydro-3H-pyrazol-3-one

[0313] At room temperature, 6 mL of TFA was added to a stirred solution of tert-butyl 5-amino-4-(4-fluorophenyl)-2-methyl-3-oxo-2,3-dihydro-1H-pyrazole-1-carboxylate (2.42 g, 7.87 mmol) in DCM (24 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (1.46 g, 89%) as a white solid. LCMS (ES, m / z): [M+H] + =208.

[0314] Step 3: N-{[4-(4-fluorophenyl)-1-methyl-5-oxo-2H-pyrazol-3-yl]carbamate}ethyl carbamate

[0315] At room temperature, ethyl N-thiocarbonylcarbamate (1.27 g, 9.70 mmol) was added to a stirred solution of 5-amino-4-(4-fluorophenyl)-2-methyl-1,2-dihydro-3H-pyrazole-3-one (1.34 g, 6.47 mmol) in DMF (13 mL). The resulting mixture was stirred at room temperature for 1 h. The reaction mixture was poured into water at 0 °C. The resulting mixture was filtered, and the filter cake was washed with water. The filtrate was concentrated under reduced pressure. This yielded the title compound (1.0 g, 46%) as a yellow solid. LCMS (ES, m / z): [M+H] + =339.

[0316] Step 4: 8-(4-fluorophenyl)-6-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4,7-dione

[0317] EtONa (402 mg, 5.91 mmol) was added to a stirred solution of N-{[4-(4-fluorophenyl)-1-methyl-5-oxo-2H-pyrazol-3-yl]aminothioyl}carbamate (1.0 g, 3.0 mmol) in EtOH (10 mL). The resulting mixture was stirred at 80 °C for 1 h. The mixture was concentrated under reduced pressure. The mixture was acidified to pH 6 with 2N HCl. The mixture was filtered, and the filter cake was washed with water. The filtrate was concentrated under reduced pressure. This yielded the title compound (872 mg, 91%) as a yellow solid. LCMS (ES, m / z): [M+H] + =293.

[0318] Step 5: 8-(4-fluorophenyl)-6-methyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4,7-dione

[0319] Proprynne bromide (65 mg, 0.55 mmol) was added to a stirred mixture of 8-(4-fluorophenyl)-6-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4,7-dione (200 mg, 0.684 mmol) and K₂CO₃ (284 mg, 2.05 mmol) in DMF (4 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 35% B to 57% B, 57% B over 8 min; wavelength: 220 nm; RT1 (min): 5.85) to give the title compound (54 mg, 24%) as a white solid. LCMS (ES, m / z): [MH] - =329. 1 H NMR (300 MHz, DMSO-d6) δ 8.45 – 8.32 (m, 2H), 7.25 – 7.11 (m, 2H), 4.07 (d, J =2.6 Hz, 2H), 3.55 (s, 3H), 3.31 (t, J = 2.5 Hz, 1H).

[0320] Synthesis method C

[0321] Example 94 - 7-chloro-8-(4-fluorophenyl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0322]

[0323] Step 1: 4-Bromo-5-chloro-2H-pyrazole-3-amine

[0324] NBS (363 mg, 2.04 mmol) was added fractionally to a stirred mixture of 5-chloro-2H-pyrazole-3-amine (120 mg, 1.02 mmol) in DCM (4 mL) at room temperature. The resulting mixture was stirred overnight at room temperature. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 0.1% FA, 10% to 100% gradient over 10 min; detector, UV 254 nm, to give the title compound (45 mg, 22%) as a yellow solid. LCMS (ES, m / z): [M+H] + =196, 198.

[0325] Step 2: 5-Chloro-4-(4-fluorophenyl)-2H-pyrazole-3-amine

[0326] At room temperature under N2, K2CO3 (169 mg, 1.22 mmol) and Pd(dtbpf)Cl2 (26 mg, 0.041 mmol) were added fractionally to a stirred mixture of 4-bromo-5-chloro-2H-pyrazole-3-amine (320 mg, 0.407 mmol) and 4-fluorophenylboronic acid (85 mg, 0.61 mmol) in dioxane (0.7 mL). The resulting mixture was stirred overnight at 100 °C under N2. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 0.1% FA, 10% to 100% gradient over 10 min; detector, UV 220 nm, to give the title compound (40 mg, 12%) as a yellow solid. LCMS (ES, m / z): [M+H] + =212.

[0327] Step 3: N-{[5-chloro-4-(4-fluorophenyl)-2H-pyrazol-3-yl]aminothioyl}carbamate

[0328] At room temperature, ethyl N-thiocarbonylcarbamate (6.2 mg, 0.047 mmol) was added to a stirred mixture of 5-chloro-4-(4-fluorophenyl)-2H-pyrazole-3-amine (50 mg, 0.047 mmol) in dimethylformamide (2 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 0.1% FA, 0% to 100% gradient over 10 min; detector, UV 254 nm, to give the title compound (10 mg, 12%) as a yellow solid. LCMS (ES, m / z): [M+H] + =343.

[0329] Step 4: 7-Chloro-8-(4-fluorophenyl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0330] Sodium ethoxide (28 mg, 0.41 mmol) was added fractionally to a stirred mixture of N-{[5-chloro-4-(4-fluorophenyl)-2H-pyrazol-3-yl]aminothioyl}carbamate (70 mg, 0.204 mmol) in ethanol (2 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 0.1% FA, 0% to 100% gradient over 10 min; detector, UV 254 nm, to give the title compound (50 mg, 83%) as a yellow solid. LCMS (ES, m / z): [M+H] + =297.

[0331] Step 5: 7-Chloro-8-(4-fluorophenyl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0332] Propylbromine (19 mg, 0.16 mmol) was added dropwise to a stirred mixture of 7-chloro-8-(4-fluorophenyl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (60 mg, 0.20 mmol) and K₂CO₃ (56 mg, 0.40 mmol) in DMF (2 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was purified by Prep-HPLC under the following conditions (column: Sunfire prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 52% B to 60% B, 60% B over 8 min; wavelength: 254 nm; RT1 (min): 6.05) to give the title compound (10 mg, 15%) as a white solid. LCMS(ES, m / z): [M+H] + = 335.1 H NMR (300 MHz, DMSO-d6) δ7.85 - 7.56 (m, 2H), 7.32 (m, 2H), 4.01 (s, 2H), 3.22 (s, 1H).

[0333] Example 95 was synthesized using synthesis method C:

[0334]

[0335] LCMS (ES, m / z): [M+H] += 379. 1H NMR (400 MHz, DMSO-d6) δ 7.95 – 7.93(m, 2H), 7.65 – 7.56 (m, 2H), 4.33 (q, 2H).

[0336] Example 96 - 8-(4-fluorophenyl)-7-hydroxy-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0337] Step 1: 5-Amino-4-(4-fluorophenyl)-1H-pyrazole-3-ol

[0338] Hydrazine hydrate (80%, 0.40 mL, 8.2 mmol) was added to a stirred solution of 2-cyano-2-(4-fluorophenyl)-N,N-dimethylacetamide (1.39 g, 6.74 mmol) in EtOH (20 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, gradient of 10% to 50% over 10 min; detector, UV 254 nm, to give the title compound (0.80 g, 61%) as a pale yellow oil. LCMS (ES, m / z): [M+H] + =194.

[0339] Step 2: N-{[4-(4-fluorophenyl)-5-hydroxy-2H-pyrazol-3-yl]carbamate}ethyl carbamate

[0340] A mixture of 5-amino-4-(4-fluorophenyl)-1H-pyrazole-3-ol (500 mg, 2.59 mmol) and ethyl N-thiocarbonylcarbamate (306 mg, 2.33 mmol) in DMF (8 mL) was stirred at room temperature for 2 h. The resulting mixture was filtered, and the filter cake was washed with water (3 x 20 mL). The solid was dried under vacuum. This yielded the title compound (230 mg, 27%) as a yellow solid. LCMS (ES, m / z): [M+H] + =325.

[0341] Step 3: 8-(4-fluorophenyl)-7-hydroxy-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0342] EtONa (46 mg, 0.68 mmol) was added to a stirred solution of N-{[4-(4-fluorophenyl)-5-hydroxy-2H-pyrazol-3-yl]aminothioyl}carbamate (110 mg, 0.339 mmol) in EtOH (2 mL) at 80 °C. The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure. The mixture was acidified to pH 6 with 2N HCl. The precipitated solid was collected by filtration and washed with water. The solid was dried under vacuum. This yielded the title compound (65 mg, 69%) as a yellow solid. LCMS (ES, m / z): [M+H] + =279.

[0343] Step 4: 8-(4-fluorophenyl)-7-hydroxy-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0344] Proprynne bromide (11 mg, 0.090 mmol) was added to a stirred solution of 8-(4-fluorophenyl)-7-hydroxy-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (50 mg, 0.18 mmol) and K₂CO₃ (75 mg, 0.54 mmol) in DMF (2.0 mL) for 2 h at room temperature. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 20% to 50% gradient over 10 min; detector, UV 254 nm, to give 76 mg of crude product. The crude product was purified by Prep-HPLC under the following conditions (column: Atlantis HILIC OBD column, 19*150 mm*5 μm; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 41% B to 49% B, 49% B over 8 min; wavelength: 220 nm; RT1 (min): 5.47), yielding the title compound (10 mg, 18%) as a white solid. LCMS (ES, m / z): [M+H] + =317. 1 H NMR (400 MHz, DMSO-d6) δ 12.89 (s, 1H), 11.93 (s, 1H), 8.25 – 8.15(m, 2H), 7.27 – 7.16 (m, 2H), 4.07 (d, J = 2.6 Hz, 2H), 3.30 – 3.27 (t, J =2.6 Hz, 1H).

[0345] Synthesis method G

[0346] Example 97-8-(4-fluorophenyl)-2-(prop-2-yn-1-ylthioalkyl)-7-(trifluoromethyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0347]

[0348] Step 1: 4-Bromo-5-(trifluoromethyl)-2H-pyrazole-3-amine

[0349] NBS (1.12 g, 6.29 mmol) was added to a stirred solution of 5-(trifluoromethyl)-2H-pyrazole-3-amine (950 mg, 6.29 mmol) in ACN (20 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure and diluted with water. The mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (500 mg, 35%) as a yellow solid. LCMS (ES, m / z): [M+H] + =230, 232.

[0350] Step 2: 4-(4-fluorophenyl)-5-(trifluoromethyl)-2H-pyrazole-3-amine

[0351] At room temperature, K₂CO₃ (721 mg, 5.22 mmol) and Pd(Dtbpf)Cl₂ (113 mg, 0.174 mmol) were added to a stirred mixture of 4-bromo-5-(trifluoromethyl)-2H-pyrazole-3-amine (400 mg, 1.74 mmol) and 4-fluorophenylboronic acid (292 mg, 2.09 mmol) in 1,4-dioxane (7 mL) and water (1 mL). The final reaction mixture was irradiated with microwave at 80 °C for 4 h. The mixture was acidified to pH 6 with 2N HCl. The resulting mixture was extracted with EA (3 x 8 mL). The combined organic layers were washed with water (8 mL) and brine (2 x 8 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 30% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (230 mg, 54%) as a yellow solid. LCMS (ES, m / z): [M+H]+ =246.

[0352] Step 3: N-{[4-(4-fluorophenyl)-5-(trifluoromethyl)-2H-pyrazol-3-yl]aminothioyl}carbamate

[0353] N-Thiocarbonylcarbamate (93 mg, 0.71 mmol) was added to a stirred solution of 4-(4-fluorophenyl)-5-(trifluoromethyl)-2H-pyrazole-3-amine (173 mg, 0.706 mmol) in DMF (4 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was poured into water (5 mL) at 0 °C. The precipitated solid was collected by filtration and washed with water (2 x 1 mL). The solid was dried under vacuum. This yielded the title compound (252 mg, 95%) as a yellow solid. LCMS (ES, m / z): [M+H] + =377.

[0354] Step 4: 8-(4-fluorophenyl)-2-thioalkyl-7-(trifluoromethyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0355] EtONa (210 mg, 3.08 mmol) was added to a stirred solution of N-{[4-(4-fluorophenyl)-5-(trifluoromethyl)-2H-pyrazol-3-yl]aminothioyl}carbamate (387 mg, 1.03 mmol) in EtOH (8 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure. The mixture was acidified to pH 6 with 2N HCl (aqueous solution). The precipitated solid was collected by filtration and washed with water (2 x 1 mL). The solid was dried under vacuum. This yielded the title compound (275 mg, 81%) as a yellow solid. LCMS (ES, m / z): [MH] - =329.

[0356] Step 5: 8-(4-fluorophenyl)-2-(prop-2-yn-1-ylthioalkyl)-7-(trifluoromethyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0357] Proprynne bromide (47 mg, 0.39 mmol) was added to a stirred mixture of 8-(4-fluorophenyl)-2-thioalkyl-7-(trifluoromethyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (130 mg, 0.394 mmol) and K₂CO₃ (163 mg, 1.18 mmol) in DMF (3 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The mixture was filtered, and the filter cake was washed with DMF (2 x 1 mL). The filtrate was purified by Prep-HPLC under the following conditions (column: Xselect CSHC18 OBD column 30*150 mm 5 μm, n; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 49% B to 61% B, 61% B over 8 min; wavelength: 254 / 220 nm; RT1 (min): 7.38), yielding the title compound (31 mg, 21%) as a grayish-white solid. LCMS (ES, m / z): [MH] - =367. 1 H NMR (300 MHz, DMSO-d6) δ7.71 – 7.60 (m, 2H), 7.39 – 7.26 (m, 2H), 3.98 (d, J = 2.6 Hz, 2H), 3.29 (t, J= 2.6 Hz, 1H).

[0358] Example 98 was synthesized using synthesis method G:

[0359]

[0360] LCMS (ES, m / z): [M+H] + = 413. 1H NMR (300 MHz, DMSO-d6) δ 7.58 – 7.53(m, 2H), 7.35 – 7.29 (m, 2H), 4.19 (q, J = 10.2 Hz, 2H).

[0361] Example 99 - 7-Methyl-8-(piperidin-1-yl)-2-[(2,2,2-trifluoroethyl)thioalkyl]-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0362]

[0363] Step 1: N-[(5-methyl-2H-pyrazol-3-yl)carbamoyl]carbamate

[0364] A solution of 5-methyl-2H-pyrazole-3-amine (1.0 g, 10 mmol) and ethyl N-thiocarbonylcarbamate (1.08 g, 8.24 mmol) in DMF (20 mL) was stirred at room temperature for 2 h. The reaction was quenched by adding ice water (50 mL) at 0 °C. The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with water (50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure, leaving the title compound (1.0 g, 43%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + = 229.

[0365] Step 2: 7-Methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0366] A solution of N-[(5-methyl-2H-pyrazol-3-yl)carbamate (1.6 g, 7.0 mmol) and EtONa (954 mg, 14.0 mmol) in EtOH (16 mL) was stirred overnight at 80 °C. The mixture was acidified to pH 6 with concentrated HCl. The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, silica gel; mobile phase, aqueous ACN, 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (1.0 g, 78%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =183.

[0367] Step 3: 7-Methyl-2-[(2,2,2-trifluoroethyl)thioalkyl]-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0368] A solution of 7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (1.0 g, 5.5 mmol), 2,2,2-trifluoroethyltrifluoromethanesulfonate (2.04 g, 8.78 mmol), and K₂CO₃ (2.28 g, 16.5 mmol) in DMF (16 mL) was stirred at room temperature for 2 h. The reaction was quenched by adding ice water (50 mL) at 0 °C. The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with water (50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (1.0 g, 69%) as a white solid. LCMS (ES, m / z): [M+H] + =265.

[0369] Step 4: 3-(methoxymethyl)-7-methyl-2-[(2,2,2-trifluoroethyl)thioalkyl]pyrazolo[1,5-a][1,3,5]triazin-4-one

[0370] A solution of 7-methyl-2-[(2,2,2-trifluoroethyl)thioalkyl]-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (1.0 g, 3.8 mmol), chloromethyl methyl ether (609 mg, 7.57 mmol), and N,N-diisopropylethylamine (1.47 g, 11.4 mmol) in DMF (20 mL) was stirred overnight at 50 °C under N2. The reaction was quenched by adding ice water (50 mL) at 0 °C. The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with water (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, silica gel; mobile phase, aqueous ACN, 10% to 100% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (1.0 g, 86%), a white solid. LCMS (ES, m / z): [M+H] + =309.

[0371] Step 5: 6-Methyl-7-nitro-2-[(2,2,2-trifluoroethyl)thioalkyl]-3H,4aH-pyrrolo[3,2-d]pyrimidin-4-one

[0372] At -20 °C, fuming HNO3 (0.17 mL, 4.1 mmol) was added dropwise to a stirred solution of 3-(methoxymethyl)-6-methyl-2-[(2,2,2-trifluoroethyl)thioalkyl]-4aH-pyrrolo[3,2-d]pyrimidin-4-one (500 mg, 1.63 mmol) in DCM (10 mL). The resulting mixture was stirred at room temperature for 2 days. The residue was alkalized to pH 7 with saturated Na2CO3 (aqueous solution). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, silica gel; mobile phase, aqueous ACN, 10% to 100% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (230 mg, 46%) as a yellow solid. LCMS (ES, m / z): [M+H] + =310.

[0373] Step 6: 8-Amino-7-methyl-2-[(2,2,2-trifluoroethyl)thioalkyl]-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0374] A solution of 7-methyl-8-nitro-2-[(2,2,2-trifluoroethyl)thioalkyl]-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (220 mg, 0.711 mmol) and Pd / C (100 mg, 45 wt%) in THF (5 mL) was stirred overnight at room temperature under a hydrogen atmosphere. The filtrate was collected by filtration and used directly for the next step without further purification. This yielded the title compound (overweight, crude), a brown liquid. LCMS (ES, m / z): [M+H] + =280.

[0375] Step 7: 7-Methyl-8-(piperidin-1-yl)-2-[(2,2,2-trifluoroethyl)thioalkyl]-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0376] At room temperature, N,N-diisopropylethylamine (278 mg, 2.15 mmol) and NaI (322 mg, 2.15 mmol) were added fractionally to a stirred solution of 8-amino-7-methyl-2-[(2,2,2-trifluoroethyl)thioalkyl]-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (0.711 mmol) and 1,5-dibromopentane (198 mg, 0.859 mmol) in THF (3 mL). The resulting mixture was stirred at 75 °C for 4 h. The reaction was quenched by adding ice water (5 mL) at 0 °C. The resulting mixture was extracted with EA (3 x 5 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 31% B to 46% B, 46% B over 6 min; wavelength: 254 / 220 nm; RT1 (min): 5.08), yielding the title compound (32 mg, 13%) as a yellow solid. LCMS (ES, m / z): [M+H] + =348. 1 H NMR (300 MHz, DMSO-d6) δ 12.50 (s, 1H), 4.21 (q, J = 10.4 Hz, 2H), 3.12 – 3.00 (m, 4H), 2.20 (s, 3H), 1.65 – 1.48 (m, 6H).

[0377] Synthesis Method J

[0378] Example 100 - 7-Methyl-8-(6-methylpyridin-3-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0379]

[0380] Step 1: 4-Bromo-5-methyl-2H-pyrazole-3-amine

[0381] NBS (18.3 g, 103 mmol) was added to a stirred solution of 5-methyl-2H-pyrazole-3-amine (10.0 g, 103 mmol) and AIBN (1.69 g, 10.3 mmol) in ACN (100 mL) at room temperature. The resulting mixture was stirred overnight at room temperature. The mixture was concentrated and diluted with water (50 mL), and extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (60 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give the title compound (overweight) as a red solid. LCMS (ES, m / z): [M+H]+ = 176.

[0382] Step 2: N-[(4-bromo-5-methyl-2H-pyrazol-3-yl)carbamate]ethyl carbamate

[0383] At room temperature, ethyl N-thiocarbonylcarbamate (14.9 g, 114 mmol) was added to a stirred solution of 103 mmol of 4-bromo-5-methyl-2H-pyrazole-3-amine in 200 mL of DMF. The resulting mixture was stirred at room temperature for 2 h. The mixture was then diluted with ice water (400 mL). The precipitated solid was collected by filtration and washed with water (3 x 50 mL). The resulting solid was dried under vacuum to give the title compound (overweight), a yellow crude solid, which could be used directly for the next step without further purification. LCMS (ES, m / z): [M+H] + =307.

[0384] Step 3: 8-Bromo-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0385] EtONa (15.5 g, 228 mmol) was added to a stirred solution of N-[(4-bromo-5-methyl-2H-pyrazol-3-yl)aminothioyl]carbamate (103 mmol) in EtOH (350 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure. The residue was dissolved in water (200 mL). The mixture was acidified to pH 6 with 2 N HCl (aqueous solution). The precipitated solid was collected by filtration and washed with water (3 x 50 mL). This yielded the title compound (24 g, 81%) as a yellow solid. LCMS (ES, m / z): [M+H] + =261.

[0386] Step 4: 8-Bromo-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one

[0387] K₂CO₃ (2.32 g, 16.8 mmol) and 4-methoxybenzyl chloride (3.50 g, 22.4 mmol) were added to a stirred solution of 8-bromo-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (1.46 g, 5.59 mmol) in DMF (20 mL) at room temperature under N₂. The resulting mixture was stirred overnight at 50 °C. The product was precipitated by adding ice water. The precipitate was collected by filtration and washed with water (3 x 50 mL). The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10 / 1). This yielded the title compound (605 mg, 22%) as a white solid. LCMS (ES, m / z): [M+H] + =501.

[0388] Step 5: 3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methyl-8-(6-methylpyridin-3-yl)pyrazolo[1,5-a][1,3,5]triazin-4-one

[0389] To a solution of 8-bromo-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one hydrobromide (300 mg, 0.515 mmol) and 6-methylpyridin-3-ylboronic acid (106 mg, 0.772 mmol) in dioxane (5 mL) and water (1 mL), K₂CO₃ (142 mg, 1.03 mmol) and Pd(dtbpf)Cl₂ (34 mg, 0.052 mmol) were added. The mixture was stirred overnight at 80 °C under N₂, and then cooled to room temperature. The resulting mixture was diluted with water (20 mL) and extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid column chromatography, eluting with ACN / water (0.05% FA) (0% to 70% over 10 minutes) to give the title compound (144 mg, 54%) as a brown solid. LCMS (ES, m / z): [M+H] + =514.

[0390] Step 6: 7-Methyl-8-(6-methylpyridin-3-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0391] A solution of 3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methyl-8-(6-methylpyridin-3-yl)pyrazolo[1,5-a][1,3,5]triazin-4-one (144 mg, 0.280 mmol) in TFA (3 mL) was stirred at 72 °C for 2 h. The resulting mixture was concentrated under vacuum to give the title compound (overweight), which could be used in the next step without further purification. LCMS (ES, m / z): [M+H] + =274.

[0392] Step 7: 7-Methyl-8-(6-methylpyridin-3-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0393] Proprynne bromide (35 mg, 0.29 mmol) was added dropwise to a stirred mixture of 7-methyl-8-(6-methylpyridin-3-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (100 mg, 0.366 mmol) and K2CO3 (152 mg, 1.10 mmol) in DMF (2 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was filtered, and the filtrate was purified by Prep-HPLC under the following conditions (column: SunFire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 5% B to 28% B, 28% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 5.08), yielding the title compound (25 mg, 22%) as a white solid. LCMS (ES, m / z): [M+H] + = 312. 1H NMR (400 MHz, DMSO-d6): δ 13.21 (s, 1H), 8.83 (d, J =2.3 Hz, 1H), 8.07 (dd, J = 8.1, 2.4 Hz, 1H), 7.37 (d, J = 8.2 Hz, 1H), 4.01(d, J = 2.6 Hz, 2H), 3.27 (t, J = 2.6 Hz, 1H), 2.51 (s, 3H), 2.49 (s, 3H).

[0394] Using synthesis method J, synthesize the examples in table J:

[0395] Table J

[0396]

[0397]

[0398]

[0399]

[0400]

[0401]

[0402] Example 109-8-[4-(hydroxymethyl)phenyl]-7-methyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0403]

[0404] Step 1: 8-[4-(hydroxymethyl)phenyl]-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazine-4-one

[0405] To a solution of 8-bromo-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one (300 mg, 0.598 mmol) and 4-(hydroxymethyl)phenylboronic acid (182 mg, 1.20 mmol) in dioxane (4 mL) and water (1 mL), K₂CO₃ (165 mg, 1.20 mmol) and Pd(dtbpf)Cl₂ (39 mg, 0.060 mmol) were added. After stirring overnight at 80 °C under N₂, the resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (180 mg, 57%) as a brown solid. LCMS (ES, m / z):[M+H] + =529.

[0406] Step 2: (4-{7-methyl-4-oxo-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-8-yl}phenyl)methyl 2,2,2-trifluoroacetate

[0407] A solution of 8-[4-(hydroxymethyl)phenyl]-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one (150 mg, 0.284 mmol) in TFA (3 mL) was stirred at 72 °C for 2 h. The resulting mixture was concentrated under vacuum to give the title compound (70 mg, 64%). The crude product was used directly in the next step without further purification. LCMS (ES, m / z): [MH] - =383

[0408] Step 3: 8-[4-(hydroxymethyl)phenyl]-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0409] At room temperature, NH3·water (1 mL) was added to a stirred mixture of (70 mg, 0.18 mmol) of methyl 2,2,2-trifluoroacetate in MeOH (5 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated under reduced pressure to give the title compound (overweight) as a grayish-white solid. LCMS (ES, m / z): [MH]- = 287.

[0410] Step 4: 8-[4-(hydroxymethyl)phenyl]-7-methyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0411] At room temperature, propargyl bromide (40 mg, 0.33 mmol) was added dropwise to a stirred mixture of 8-[4-(hydroxymethyl)phenyl]-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (0.182 mmol) and K₂CO₃ (173 mg, 1.25 mmol) in DMF (3 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was filtered, and the filtrate was purified by Prep-HPLC under the following conditions (2#SHIMADZU(HPLC-01)): column, Sunfire prep C18 column, 30*150 mm, 5 µm; mobile phase, water (0.1% FA) and ACN (23% ACN increased to 37% within 7 min), to give the title compound (22 mg, 37%) as a white solid. LCMS (ES, m / z): [M+H] + =327. 1 H NMR (400 MHz, DMSO-d6): δ 12.94(s, 1H), 7.71 (d, J = 8.1 Hz, 2H), 7.39 (d, J = 8.0 Hz, 2H), 5.19 (s, 1H),4.53 (s, 2H), 4.01 (d, J = 2.7 Hz, 2H), 3.28 (t, J = 2.7 Hz, 1H), 2.46 (s,3H).

[0412] Synthesis method K

[0413] Example 141-8-(4-fluorophenyl)-7-methyl-2-[(2,2,3,3-tetrafluoropropyl)thioalkyl]-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0414]

[0415] Step 1: Ethyl 2-(4-fluorophenyl)-3-oxobutyronitrile

[0416] NaH (60% in mineral oil) (5.50 g, 229 mmol) was added to a stirred solution of 10 g (74 mmol) of 4-fluorophenylacetonitrile (THF) in 50 mL of THF at 0 °C under N2. The resulting mixture was stirred at 0 °C under N2 for 1 h. EA (26.1 g, 296 mmol) was added dropwise to the mixture at 0 °C. The resulting mixture was stirred again overnight at room temperature. The mixture was acidified to pH 5 with HCl (aqueous solution). The resulting mixture was concentrated under reduced pressure. The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was used directly for the next step without further purification. This yielded the title compound (11 g, 84%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =178.

[0417] Step 2: 4-(4-fluorophenyl)-5-methyl-2H-pyrazole-3-amine

[0418] At room temperature, hydrazine hydrate (80%; 5.46 g, 109 mmol) was added to a solution of 2-(4-fluorophenyl)-3-oxobutyronitrile (9.67 g, 54.6 mmol) in EtOH (100 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure. The residue was dissolved in EA (100 mL). The organic layer was washed with water (3 x 80 mL) and brine (150 mL) and dried over Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The crude product could be used directly for the next step without further purification. This yielded the title compound (7.46 g, 72%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =192.

[0419] Step 3: N-[[4-(4-fluorophenyl)-5-methyl-2H-pyrazol-3-yl]carbamoyl]carbamate

[0420] N-Thiocarbonylcarbamate (3.43 g, 26.1 mmol) was added dropwise to a stirred mixture of 4-(4-fluorophenyl)-5-methyl-2H-pyrazole-3-amine (5.00 g, 26.1 mmol) in DMF (40 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by adding water (50 mL) at 0 °C. The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with water (50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (6.0 g, 71%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =323.

[0421] Step 4: 8-(4-fluorophenyl)-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0422] A solution of N-{[4-(4-fluorophenyl)-5-methyl-2H-pyrazol-3-yl]aminothioyl}carbamate (7.0 g, 22 mmol) and sodium ethoxide (4.43 g, 65.1 mmol) in ethanol (70 mL) was stirred at 80 °C for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in water (100 mL). The residue was neutralized to pH 6 with concentrated HCl. The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with water (50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (4.4 g, 73%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =277.

[0423] Step 5: 8-(4-fluorophenyl)-7-methyl-2-[(2,2,3,3-tetrafluoropropyl)thioalkyl]-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0424] At 0 °C under N2, DIAD (146 mg, 0.724 mmol) and PPh3 (190 mg, 0.724 mmol) were added dropwise to a stirred solution of 8-(4-fluorophenyl)-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (100 mg, 0.362 mmol) and 2,2,3,3-tetrafluoropropanol (72 mg, 0.54 mmol) in THF (2 mL). The resulting mixture was stirred at 65 °C under N2 for 5 h. The mixture was concentrated under reduced pressure. The mixture was extracted with EA (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 49% B to 59% B, 59% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 5.50), yielding the title compound (48 mg, 34%) as a white solid. LCMS (ES, m / z): [M+H] + =391. 1H NMR (300MHz, DMSO-d6) δ 13.15 (s, 1H), 7.89 – 7.50 (m, 2H), 7.44 – 7.04 (m, 2H), 6.60 (tt, J = 52.3, 4.0 Hz, 1H), 4.08 (t, J = 17.5 Hz, 2H), 2.46 (s, 3H).

[0425] The examples in Table K were synthesized using synthesis method K:

[0426] Table K

[0427]

[0428]

[0429] Example 152-8-(4-fluorophenyl)-2-[(5-hydroxypent-2-yn-1-yl)thioalkyl]-7-methyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0430]

[0431] Step 1: 8-(4-fluorophenyl)-7-methyl-2-{[5-(oxacyclohexane-2-yloxy)pent-2-yn-1-yl]thioalkyl}-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0432] DIAD (220 mg, 1.09 mmol) was added dropwise to a stirred solution of 5-(oxacyclohexane-2-yloxy)pentan-2-yn-1-ol (synthesized using the procedure described in Org. Biomol. Chem., 2011, 9, 8000-8002; 100 mg, 0.543 mmol), 8-(4-fluorophenyl)-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (180 mg, 0.652 mmol) and PPh3 (285 mg, 1.09 mmol) in THF (2 mL). The resulting mixture was stirred at 65 °C under N2 for 2 h. The resulting mixture was diluted with water (5 mL). The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was dissolved in ACN (3 mL). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN (0.05% FA), 20% to 70% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (31 mg, 13%) as a white solid. LCMS (ES, m / z): [MH] - =441.

[0433] Step 2: 8-(4-fluorophenyl)-2-[(5-hydroxypent-2-yn-1-yl)thioalkyl]-7-methyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0434] A solution of 8-(4-fluorophenyl)-7-methyl-2-{[5-(oxacyclohexane-2-yloxy)pent-2-yn-1-yl]thioalkyl}-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (31 mg, 0.070 mmol) and 2N hydrogen chloride (2 mL) was stirred at room temperature for 2 h. The residue was neutralized to pH 7 with saturated NaHCO3 (aqueous solution). The resulting mixture was extracted with EA (3 x 5 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 33% B to 45% B, 45% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 6.38), yielding the title compound (4.5 mg, 18%) as a white solid. LCMS (ES, m / z): [M+H] + =359. 1H NMR (300 MHz, DMSO-d6) δ 12.9 (s,1H), 7.84-.76 (m,2H), 7.30-7.24 (m, 2H), 4.80 (s, 1H), 3.99 (t, J = 2.4 Hz, 2H), 3.45 (m, 2H), 2.45 (s, 3H), 2.38 – 2.27 (m, 2H).

[0435] Example 153-2-[(4-fluorobut-2-yn-1-yl)thioalkyl]-8-(4-fluorophenyl)-7-methyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0436]

[0437] Step 1: 8-(4-fluorophenyl)-2-[(4-hydroxybut-2-yn-1-yl)thioalkyl]-7-methyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0438] PPh3 (380 mg, 7.24 mmol) was added to a stirred mixture of 8-(4-fluorophenyl)-7-methyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (200 mg, 3.62 mmol) and butynediol (93 mg, 5.4 mmol) in THF (4 mL) at room temperature. DIAD (292 mg, 7.24 mmol) was added dropwise to the mixture at 0 °C under N2. The resulting mixture was stirred at room temperature for another 2 h. The resulting mixture was concentrated under reduced pressure. Water (5 mL) was added to the mixture. The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, silica gel; mobile phase, aqueous ACN solution, 10% to 40% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (90 mg, 36%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =345.

[0439] Step 2: 8-(4-fluorophenyl)-2-[(4-hydroxybut-2-yn-1-yl)thioalkyl]-3-[(4-methoxyphenyl)methyl]-7-methylpyrazolo[1,5-a][1,3,5]triazine-4-one

[0440] At room temperature, TEA (134 mg, 1.32 mmol) and 4-methoxybenzyl chloride (175 mg, 1.12 mmol) were added fractionally to a stirred solution of 8-(4-fluorophenyl)-2-[(4-hydroxybut-2-yn-1-yl)thioalkyl]-7-methyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (350 mg, 1.02 mmol) in DMF (7 mL). The resulting mixture was stirred at 100 °C for 2 h. Water (10 mL) was added to the mixture. The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with water (2 x 10 mL) and brine (5 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, ACN aqueous solution, 30% to 60% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (103 mg, 22%) as a yellow solid. LCMS (ES, m / z): [M+H] + =465.

[0441] Step 3: 2-[(4-fluorobut-2-yn-1-yl)thioalkyl]-8-(4-fluorophenyl)-3-[(4-methoxyphenyl)methyl]-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one

[0442] DAST (110 mg, 0.684 mmol) was added dropwise to a stirred solution of 8-(4-fluorophenyl)-2-[(4-hydroxybut-2-yn-1-yl)thioalkyl]-3-[(4-methoxyphenyl)methyl]-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one (106 mg, 0.228 mmol) in DCM (2 mL). The resulting mixture was stirred overnight at room temperature under N2. The reaction was quenched with saturated sodium bicarbonate aqueous solution at 0 °C. The resulting mixture was extracted with DCM (3 x 3 mL). The combined organic layers were washed with brine (2 x 3 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, ACN aqueous solution, 35% to 70% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (33 mg, 31%), as a yellow solid. LCMS (ES, m / z): [M+H] + =467.

[0443] Step 4: 2-[(4-fluorobut-2-yn-1-yl)thioalkyl]-8-(4-fluorophenyl)-7-methyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0444] At room temperature, a stirred solution of 2-[(4-fluorobut-2-yn-1-yl)thioalkyl]-8-(4-fluorophenyl)-3-[(4-methoxyphenyl)methyl]-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one (33 mg, 0.071 mmol) was added to TFA (1 mL). The resulting mixture was stirred at 70 °C for 4 h. The mixture was concentrated under reduced pressure. THF (3 mL) was added to the mixture. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the crude product. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 45% B to 59% B, 59% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 5.55), yielding the title compound (3.6 mg, 15%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =347. 1 H NMR (300MHz, DMSO-d6) δ 7.78 (dd, J = 8.6, 5.6 Hz, 2H), 7.26 (t, J = 8.8 Hz, 2H), 5.25-5.00 (d, J = 47.1 Hz, 2H), 4.11 (d, J = 7.4 Hz, 2H), 2.45 (s, 3H).

[0445] Example 154-8-(4-fluorophenyl)-7-(hydroxymethyl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0446]

[0447] Step 1: Ethyl 5-amino-4-bromo-1H-pyrazole-3-carboxylate

[0448] NBS (6.31 g, 35.5 mmol) was added to a stirred solution of ethyl 5-amino-1H-pyrazole-3-carboxylate (5.0 g, 32 mmol) in 50 mL of ACN at room temperature. The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1), to give the title compound (6.0 g, 80%) as a brown oil. LCMS (ES, m / z): [M+H] + =234.

[0449] Step 2: (5-amino-4-bromo-1H-pyrazole-3-yl)methanol

[0450] LiBH4 (20.5 mL, 41 mmol, 2M solution in THF) was added dropwise to a stirred solution of ethyl 5-amino-4-bromo-1H-pyrazole-3-carboxylate (6.0 g, 34 mmol) in 100 mL of THF. The resulting mixture was stirred overnight at 60 °C under N2. The mixture was then cooled to room temperature. The reaction was quenched with saturated NH4Cl (aqueous solution) at 0 °C. The resulting mixture was diluted with 100 mL of water. The resulting mixture was extracted with EA (3 x 200 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1), to give the title compound (3.6 g, 73%) as a brown oil. LCMS (ES, m / z): [M+H] + =192.

[0451] Step 3: [5-amino-4-(4-fluorophenyl)-1H-pyrazole-3-yl]methanol

[0452] At room temperature under N2, Pd(dtbpf)Cl2 (204 mg, 0.313 mmol), KHCO3 (626 mg, 6.25 mmol), and 4-fluorophenylboronic acid (874 mg, 6.25 mmol) were added to a stirred solution of (5-amino-4-bromo-1H-pyrazol-3-yl)methanol (600 mg, 3.13 mmol) in dioxane (6 mL) and water (0.6 mL). The resulting mixture was stirred overnight at 85 °C under N2. The mixture was then cooled to room temperature. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 5 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions (column, C18; mobile phase A: water (0.1% NH3·H2O), mobile phase B: ACN, 5% to 50% gradient over 10 min; detector, UV 254 nm), yielding the title compound (150 mg, 23.08%) as an orange solid. LCMS (ES, m / z): [M+H] + =208.

[0453] Step 4: N-{[4-(4-fluorophenyl)-5-(hydroxymethyl)-2H-pyrazol-3-yl]carbamate}ethyl carbamate

[0454] Ethyl N-thiocarbonylcarbamate (125 mg, 0.956 mmol) was added to a stirred solution of [5-amino-4-(4-fluorophenyl)-1H-pyrazol-3-yl]methanol (180 mg, 0.869 mmol) in DMF (2 mL). The resulting mixture was stirred at room temperature under N2 for 2 h. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 5 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give the title compound (200 mg, 68%) as an orange solid. LCMS (ES, m / z): [M+H] + =339.

[0455] Step 5: 8-(4-fluorophenyl)-7-(hydroxymethyl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0456] Sodium ethoxide (121 mg, 1.77 mmol) was added in portions to a stirred solution of N-{[4-(4-fluorophenyl)-5-(hydroxymethyl)-2H-pyrazol-3-yl]aminothioyl}carbamate (200 mg, 0.591 mmol) in EtOH (4 mL). The resulting mixture was stirred at 80 °C under N2 for 2 h. The mixture was then cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (5 mL). The mixture was acidified to pH 5 with concentrated HCl. The precipitated solid was collected by filtration and washed with water (2 x 2 mL). The resulting solid was dried under vacuum to give the title compound (150 mg, 87%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =293.

[0457] Step 6: 8-(4-fluorophenyl)-7-(hydroxymethyl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0458] Proprynne bromide (16 mg, 0.14 mmol) and K₂CO₃ (71 mg, 0.51 mmol) were added to a stirred solution of 8-(4-fluorophenyl)-7-(hydroxymethyl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (50 mg, 0.17 mmol) in DMF (2 mL) at room temperature under N₂. The resulting mixture was stirred at room temperature under N₂ for 2 h. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 5 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 28% B to 52% B, 52% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 6.03), yielding the title compound (12.7 mg, 22%) as a white solid. LCMS (ES, m / z): [M+H] +=331. 1H NMR (400 MHz, DMSO-d6) δ 13.06 (s, 1H), 8.06-7.97 (m, 2H), 7.23-7.29 (m, 2H), 5.56 (s, 1H), 4.59 (s, 2H), 4.02 (d, J = 2.6 Hz, 2H), 3.28 (t, J = 2.6 Hz, 1H).

[0459] Example 155-7-(difluoromethyl)-8-(4-fluorophenyl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0460]

[0461] Step 1: 8-(4-fluorophenyl)-4-oxo-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxaldehyde

[0462] At room temperature under N2, Dess-Martin periodane (231 mg, 0.546 mmol) was added to a stirred solution of 8-(4-fluorophenyl)-7-(hydroxymethyl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (60 mg, 0.18 mmol) in THF (2 mL). The resulting mixture was stirred overnight at room temperature under N2. The reaction was quenched by adding saturated NaHCO3 (aqueous solution) (5 mL) at 0 °C. The resulting mixture was extracted with EA (3 x 3 mL). The combined organic layers were washed with brine (2 x 3 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give the title compound (50 mg, 84%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =329.

[0463] Step 2: 7-(difluoromethyl)-8-(4-fluorophenyl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0464] DAST (74 mg, 0.46 mmol) was added to a stirred solution of 8-(4-fluorophenyl)-4-oxo-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxaldehyde (50 mg, 0.15 mmol) in DCM (2 mL). The resulting mixture was stirred overnight at room temperature under N2. The reaction was quenched by adding saturated NaHCO3 (aqueous solution) (5 mL) at 0 °C. The resulting mixture was extracted with DCM (3 x 5 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: XBridge Shield RP18 OBD column, 19*150 mm, 5 μm; mobile phase A: water (0.05% NH3H2O), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 19% B to 47% B, 47% B over 7 min; wavelength: 220 nm; RT1 (min): 5.63), yielding the title compound 7 (8.4 mg, 16%) as a pale yellow solid. LCMS (ES, m / z): [M+H]+ =351. 1HNMR (400 MHz, DMSO-d6) δ 7.84 – 7.75 (m, 2H), 7.31 – 7.19 (m, 2H), 6.97 (t,1H), 3.89 (d, J = 2.7 Hz, 2H), 3.15 (t, J = 2.6 Hz, 1H).

[0465] Synthesis method E

[0466] Example 156-8-(4-fluorophenyl)-7-(morpholin-2-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0467]

[0468] Step 1: 2-[2-cyano-2-(4-fluorophenyl)acetyl]morpholine-4-carboxylic acid tert-butyl ester

[0469] At 0 °C, NaH (1.49 g, 37.2 mmol, 60% in mineral oil) was added to a stirred solution of 2.7 g (20 mmol) of 4-fluoro-phenylacetonitrile (THF) in 50 mL of THF. At 0 °C, 4-tert-butyl-2-methylmorpholine-2,4-dicarboxylate (7.35 g, 30 mmol) was added to the mixture. The resulting mixture was stirred overnight at room temperature. The reaction was quenched with saturated NH4Cl (aqueous solution) (30 mL) at room temperature. The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with brine (20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1) to give the title compound (6.7 g, 96%) as a yellow oil. LCMS (ES, m / z): [M-Boc+H] + =249.

[0470] Step 2: 2-[5-amino-4-(4-fluorophenyl)-1H-pyrazol-3-yl]morpholine-4-carboxylic acid tert-butyl ester

[0471] At room temperature, hydrazine hydrate (80%) (135 mg, 2.16 mmol) was added to a stirred solution of 2-[2-cyano-2-(4-fluorophenyl)acetyl]morpholine-4-carboxylic acid tert-butyl ester (3.48 g, 10 mmol) in EtOH (60 mL). The resulting mixture was stirred at 45 °C for 2 h. The mixture was then cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (538 mg, 15%) as a yellow oil. LCMS (ES, m / z): [MH] - =361.

[0472] Step 3: 2-[5-({[(ethoxycarbonyl)amino]methionyl}amino)-4-(4-fluorophenyl)-1H-pyrazol-3-yl]morpholine-4-carboxylic acid tert-butyl ester

[0473] At room temperature, tert-butyl N-thiocarbonylcarbamate (136 mg, 1.04 mmol) was added dropwise to a stirred solution of 2-[5-amino-4-(4-fluorophenyl)-1H-pyrazol-3-yl]morpholino-4-carboxylate (538 mg, 1.49 mmol) in DMF (3 mL). The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched by adding 50 mL of water at 0 °C. The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with water (20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to give the title compound (898 mg, overweight) as a yellow oil. LCMS (ES, m / z): [M+H] + 494.

[0474] Step 4: 2-[8-(4-fluorophenyl)-4-oxo-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazin-7-yl]morpholine-4-carboxylic acid tert-butyl ester

[0475] Sodium ethoxide (205 mg, 3.01 mmol) was added fractionally to a stirred solution of 2-[5-({[(ethoxycarbonyl)amino]meththioyl}amino)-4-(4-fluorophenyl)-1H-pyrazol-3-yl]morpholino-4-carboxylic acid tert-butyl ester (742 mg, 1.50 mmol) in EtOH (7 mL). The resulting mixture was refluxed at 80 °C for 1 h. The residue was filtered off, and the precipitate was dissolved in water (40 mL) and acidified to pH 7 with 2N HCl. The resulting mixture was extracted with EA (3 x 15 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to give the title compound (360 mg, 54%) as a yellow oil. LCMS (ES, m / z): [M+H] + =392.

[0476] Step 5: 2-[8-(4-fluorophenyl)-4-oxo-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-7-yl]morpholine-4-carboxylic acid tert-butyl ester

[0477] At room temperature, propargyl bromide (48 mg, 0.41 mmol) was added dropwise to a stirred mixture of 2-[8-(4-fluorophenyl)-4-oxo-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazin-7-yl]morpholino-4-carboxylic acid tert-butyl ester (304 mg, 0.679 mmol) and K2CO3 (282 mg, 2.04 mmol) in DMF (2 mL). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was filtered and purified by Prep-HPLC under the following conditions (2#SHIMADZU(HPLC-01)): column, XBridgePrep C18 OBD column, 30*50 mm, 5µm 13nm; mobile phase, water (10 mmol / L NH4HCO3) and ACN (15% ACN, increased to 40% in 8 min), to give the title compound (70 mg, 21%) as a white solid. LCMS (ES, m / z): [MH] - =484.

[0478] Step 6: 8-(4-fluorophenyl)-7-(morpholin-2-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0479] A solution of tert-butyl 2-[8-(4-fluorophenyl)-4-oxo-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-7-yl]morpholin-4-carboxylate (44 mg, 0.091 mmol) in DCM (4 mL) and TFA (2 mL) was stirred for 2 h at room temperature and concentrated under vacuum to give the crude product. The crude product was purified by Prep-HPLC under the following conditions (2#SHIMADZU(HPLC-01)): column, Sunfire prep C18 column, 30*150 mm, 5 µm; mobile phase, water (0.1% FA) and ACN (17% ACN increased to 32% within 7 min), to give the title compound (13 mg, 37%) as a white solid. LCMS (ES, m / z): [M+H] + =386. 1H NMR (400 MHz, DMSO-d6) δ 9.05 (s, 1H), 7.79 – 7.70 (m,2H), 7.31 – 7.22 (m, 2H), 4.90 (d, J = 10.4 Hz, 1H), 4.14 – 4.06 (m, 1H), 3.93 (s, 1H), 3.97 – 3.83 (m, 2H), 3.83 – 3.70 (m, 1H), 3.52 (d, J = 12.6 Hz, 1H), 3.35 – 3.28 (m, 2H), 3.09 (t, J = 2.6 Hz, 1H).

[0480] The embodiments in Table E were synthesized using synthesis method E:

[0481] Table E

[0482]

[0483] Racemic 8-(4-chlorophenyl)-7-(morpholin-2-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (500 mg) was synthesized using synthetic method E. Enantiomers were separated by SFC under the following conditions (column: N--(S, S)-Whelk-O1 (Ser#76233), 4.6*100 mm, 3.5 μm; mobile phase B: MeOH (20 mMNH3); flow rate: 3 mL / min; gradient: isocratic 40% B; wavelength: 220 nm; injection volume: 3 mL) to obtain enantiomer 1 (130 mg) and enantiomer 2 (60 mg). These compounds were purified by Prep-HPLC under the following conditions (column: Sunfire prep C18 column, 30*150mm, 5μm; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 20% B to 38% B over 8 min; wavelength: 254 / 220 nm; RT1 (min): 6.27), yielding the title compound TFA salt (Example 159, enantiomer 1, 130 mg), as a grayish-white solid. LCMS (ES, m / z): [M+H]+ = 402. ¹H NMR (400 MHz, DMSO-d6) δ 13.35 (s, 1H), 9.21 (s, 1H), 9.02 (s, 1H), 7.79 (d, J = 13.8 Hz, 2H), 7.55 (d, J = 12.4, 11.2 Hz, 2H), 4.95 – 4.93 (m, 1H), 4.13 – 3.91 (m, 4H), 3.62 – 3.51 (m, 3H), 3.30 – 3.26 (m, 3H) and the title compound TFA salt (Example 160, enantiomer 2, 60 mg), as a pale yellow solid. LCMS (ES, m / z): [M+H]+ =402. 1H NMR (400 MHz, DMSO-d6) δ 13.25(s, 1H), 9.22 (s, 1H), 9.03 (s, 1H), 7.79 (d, J = 8.3 Hz, 2H), 7.55 (d, J =8.3 Hz, 2H), 4.95 – 4.93 (dd, J = 10.8, 2.4 Hz, 1H), , 4.19 – 3.91 (m, 4H), 3.62 – 3.39 (m, 3H), 3.33 – 3.23 (m, 3H).

[0484] Example 161-8-(4-fluorophenyl)-7-(4-methylmorpholin-2-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0485]

[0486] At 0 °C and N2, HCHO (95 mg, 1.12 mmol, 37 wt% solution in H2O) was added to a stirred solution of 8-(4-fluorophenyl)-7-(morpholin-2-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (Example 156, 150 mg, 0.389 mmol) in ACN (3 mL). The resulting mixture was stirred at room temperature for 1 h. NaBH3CN (73 mg, 1.2 mmol) was added to the mixture at room temperature. The resulting mixture was stirred again overnight at room temperature. The resulting mixture was quenched with 2M HCl (0.5 mL). The resulting mixture was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), in a 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the crude product. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 13% B to 37% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 5.40), yielding the title compound (19.7 mg, 13%) as a white solid. LCMS (ES, m / z): [M+H] + =400. 1 H-NMR (400 MHz, DMSO-d6) δ 10.43 (s, 1H), 7.77 – 7.74 (m, 2H), 7.28 – 7.23 (m, 2H), 4.78 (d, J = 10.3 Hz, 1H), 4.10 (d, J = 12.3 Hz, 1H), 3.92 – 3.82 (m, 3H), 3.53 – 3.50 (m, 2H), 3.32 –3.29 (m, 1H), 3.12 (t, J =2.6 Hz, 1H), 2.99 – 2.98 (m, 1H), 2.77 (s, 3H).

[0487] Synthesis Method X

[0488] Example 162-8-(4-fluorophenyl)-7-(4-isopropylmorpholin-2-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0489]

[0490] At room temperature, 2-iodopropane (26.5 mg, 0.156 mmol) was added to a stirred mixture of 8-(4-fluorophenyl)-7-(morpholin-2-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (Example 156, 50 mg, 0.13 mmol) and K2CO3 (54 mg, 0.39 mmol) in DMF (2 mL). The resulting mixture was stirred overnight at 65 °C. The resulting mixture was filtered, and the filter cake was washed with MeOH (3 x 2 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), in a 10% to 80% gradient over 10 min; detector, UV 254 nm, to give the crude product. The crude product was purified by Prep-HPLC under the following conditions (column: XBridgeShield RP18 OBD column, 19*150 mm, 5 μm; mobile phase A: water (0.05% NH3)). . H2O), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 19% B to 40% B over 7 min; wavelength: 220 nm; RT1 (min): 5.91), yielded the title compound (5.4 mg, 10%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =428. 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 9.98 (s, 1H), 7.76 – 7.72 (m, 2H), 7.29 – 7.25 (m, 2H), 4.90 (d, J = 10.3 Hz, 1H), 4.17 (d, J = 12.5 Hz, 1H), 3.95 – 3.87 (m, 3H), 3.58 – 3.50 (m, 2H), 3.44 – 3.35 (m, 2H), 3.13 (septet, J = 6.8 Hz, 1H), 3.12 – 3.11 (m, 1H), 1.25 (d, J = 6.5 Hz, 6H).

[0491] Examples from Synthesis Table X using Synthesis Method X:

[0492] Table X

[0493]

[0494] Synthesis method F

[0495] Examples 165 and 166 - 8-(4-fluorophenyl)-7-[oxacyclopentan-3-yl]-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0496]

[0497] Step 1: 2-(4-fluorophenyl)-3-oxo-3-(oxacyclopentan-3-yl)propionitrile

[0498] NaH (0.71 g, 30 mmol, 60% in mineral oil) was added to a stirred solution of 4-fluorophenylacetonitrile (2.0 g, 15 mmol) in THF (20 mL) at 0 °C under N2. The resulting mixture was stirred at 0 °C under N2 for 30 min. Methyl oxacyclopentane-3-carboxylate (3.85 g, 29.6 mmol) was added to the mixture at 0 °C. The resulting mixture was stirred again overnight at room temperature. The mixture was acidified to pH 6 with 2N HCl (aqueous solution). The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (2 x 20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (8:1) to give the title compound (2.74 g, 79%) as a yellow oil. LCMS (ES, m / z): [MH] - =232.

[0499] Step 2: 4-(4-fluorophenyl)-5-(oxacyclopentan-3-yl)-2H-pyrazole-3-amine

[0500] Hydrazine hydrate (7.51 g, 120 mmol, 80 wt%) was added to a stirred solution of 2-(4-fluorophenyl)-3-oxo-3-(oxepane-3-yl)propionitrile (14 g, 60 mmol) in EtOH (31 mL). The resulting mixture was stirred overnight at 80 °C. The mixture was concentrated under reduced pressure. The residue was dissolved in water (100 mL). The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, ACN aqueous solution (0.05% NH₃H₂O), in a 30% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (1.13 g, 8%) as a yellow solid. LCMS (ES, m / z): [M+H] + =248.

[0501] Step 3: N-{[4-(4-fluorophenyl)-5-(oxacyclopentan-3-yl)-2H-pyrazol-3-yl]aminothioyl}carbamate

[0502] At room temperature, ethyl N-thiocarbonylcarbamate (530 mg, 4.04 mmol) was added to a stirred solution of 4-(4-fluorophenyl)-5-(oxacyclopentan-3-yl)-2H-pyrazole-3-amine (1.0 g, 4.0 mmol) in DMF (10 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was poured into water (30 mL) at 0 °C. The precipitated solid was collected by filtration and washed with water (2 x 10 mL). The solid was dried under vacuum. This yielded the title compound (1.2 g, 78%) as a yellow solid. LCMS (ES, m / z): [M+H]+ = 379.

[0503] Step 4: 8-(4-fluorophenyl)-7-(oxacyclopentan-3-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0504] Sodium ethoxide (432 mg, 6.34 mmol) was added to a stirred solution of N-{[4-(4-fluorophenyl)-5-(oxacyclopentan-3-yl)-2H-pyrazol-3-yl]aminothioyl}carbamate (1.2 g, 3.2 mmol) in EtOH (12 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure. The mixture was acidified to pH 6 with 2N HCl (aqueous solution). The precipitated solid was collected by filtration and washed with water (2 x 5 mL). The solid was dried under vacuum. This yielded the title compound (1.08 g, 87%) as a yellow solid. LCMS (ES, m / z): [M+H]+ = 333.

[0505] Step 5: 8-(4-fluorophenyl)-7-[oxacyclopentan-3-yl]-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (enantiomer 1 = Example 165; enantiomer 2 = Example 166)

[0506] At room temperature, K₂CO₃ (1.35 g, 9.75 mmol) and propargyl bromide (0.31 g, 2.6 mmol) were added fractionally to a stirred solution of 8-(4-fluorophenyl)-7-(oxacyclopentan-3-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (1.08 g, 3.25 mmol) in DMF (11 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was filtered, and the filter cake was washed with DMF (3 x 2 mL). The filtrate was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, ACN aqueous solution, 30% to 70% gradient over 10 min; detector, UV 254 nm. This produced 8-(4-fluorophenyl)-7-(oxacyclopentan-3-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (540 mg), a yellow solid. The product was purified by SFC under the following conditions: (Column: CHIRALPAK IG, 3*25 cm, 5 μm; Mobile phase A: CO2, Mobile phase B: IPA-HPLC; Flow rate: 60 mL / min; Gradient: isocratic 35% B; Column temperature (°C): 35; Back pressure (bar): 100; Wavelength: 206 nm; RT1 (min): 7.22; RT2 (min): 9.35; Sample solvent: MeOH:DCM=1:1-HPLC; Injection volume: 1.6 mL; Number of runs: 14). Two separate enantiomers were obtained, which were further purified by prep-HPLC under the following conditions: (Column: XBridge Shield RP18 OBD column, 19*150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 25 mL / min; Gradient: 19% B to 45% B over 8 min, 45% B B; wavelength: 220 nm; RT1 (min): 6.18), yielding the title compound (enantiomer 1: 206 mg, 17%). LCMS (ES, m / z): [M+H] + = 371. 1¹H NMR (400 MHz, DMSO-d⁶) δ 13.02 (s, 1H), 7.74–7.64 (m, 2H), 7.34–7.23 (m, 2H), 4.08–3.99 (m, 1H), 3.97 (d, J = 2.7 Hz, 2H), 3.87 (td, J = 8.1, 5.6 Hz, 1H), 3.83–3.70 (m, 3H), 3.27 (t, J = 2.6 Hz, 1H), 2.31–2.11 (m, 2H) and the title compound (enantiomer 2: 208 mg, 17%), as a grayish-white solid. LCMS (ES, m / z): [M+H] + = 371. 1 H NMR (400 MHz, DMSO-d6) δ 13.02 (s, 1H), 7.74 – 7.64 (m, 2H), 7.34 –7.23 (m, 2H), 4.08 – 3.99 (m, 1H), 3.97 (d, J = 2.7 Hz, 2H), 3.87 (td, J =8.1, 5.6 Hz, 1H), 3.83 – 3.70 (m, 3H), 3.27 (t, J = 2.6 Hz, 1H), 2.31 – 2.11 (m, 2H).

[0507] The examples in Table F were synthesized using synthesis method F:

[0508] Table F

[0509]

[0510]

[0511] Examples 173 and 174 - 8-(4-fluorophenyl)-7-[oxacyclohexane-2-yl]-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0512]

[0513] Step 1: 2-(4-fluorophenyl)-3-(oxacyclohexane-2-yl)-3-oxopropionitrile

[0514] A solution of 4-fluoro-phenylacetonitrile (51 g, 377 mmol) in THF (500 mL) was treated with NaH (18.1 g, 755 mmol) for 30 min at 0 °C under N2, followed by the addition of methyl oxacyclohexane-2-carboxylate (65.3 g, 453 mmol) at 0 °C. The resulting mixture was stirred overnight at room temperature. The mixture was acidified to pH 6 with 2N HCl. The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (9:1), to give the title compound (90 g, 96%) as a yellow oil. LCMS (ES, m / z): [M+H] + =248.

[0515] Step 2: 4-(4-fluorophenyl)-5-(oxacyclohexane-2-yl)-2H-pyrazole-3-amine

[0516] At room temperature, 80% hydrazine hydrate (23.3 g, 465 mmol) and TFA (42.4 g, 372 mmol) were added to a stirred solution of 2-(4-fluorophenyl)-3-(oxacyclohexane-2-yl)-3-oxopropionitrile (92 g, 372 mmol) in EtOH (900 mL). The resulting mixture was stirred at 60 °C for 2 h. The mixture was concentrated under reduced pressure. The residue was dissolved in water. The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The crude title compound could be used directly for the next step without further purification. LCMS (ES, m / z): [M+H] + =262.

[0517] Step 3: N-{[4-(4-fluorophenyl)-5-(oxacyclohexane-2-yl)-2H-pyrazol-3-yl]carbamate}ethyl carbamate

[0518] At room temperature, ethyl N-thiocarbonylcarbamate (75.3 g, 574 mmol) was added to a stirred solution of 150 g (574 mmol) of 4-(4-fluorophenyl)-5-(oxacyclohexane-2-yl)-2H-pyrazole-3-amine in DMF (500 mL). The resulting mixture was stirred at room temperature for 2 h. The product was precipitated by adding water / ice (4 L). The precipitated solid was collected by filtration, washed with water (4 x 100 mL), and dried under reduced pressure. This yielded the title compound (200 g, 89%) as a yellow solid. LCMS (ES, m / z): [M+H] + =393.

[0519] Step 4: 8-(4-fluorophenyl)-7-(oxacyclohexane-2-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0520] Sodium ethoxide (104 g, 1529 mmol) was added to a stirred solution of N-{[4-(4-fluorophenyl)-5-(oxacyclohexane-2-yl)-2H-pyrazol-3-yl]carbamate (200 g, 510 mmol) in EtOH (1.3 L). The resulting mixture was stirred at 80 °C for 3 h. The mixture was concentrated under reduced pressure. The residue was dissolved in water (500 mL). The mixture was acidified to pH 6 with 2N HCl. The precipitated solid was collected by filtration and washed with water (2 x 40 mL). This yielded the title compound (110 g, 62%) as a yellow solid. LCMS (ES, m / z): [M+H] + =347.

[0521] Step 5: 8-(4-fluorophenyl)-7-(oxacyclohexane-2-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (Examples 173 and 174)

[0522] At room temperature, K₂CO₃ (29.9 g, 217 mmol) and propargyl bromide (6.87 g, 57.7 mmol) were added fractionally to a stirred solution of 8-(4-fluorophenyl)-7-(oxacyclohexan-2-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (25 g, 72 mmol) in DMF (250 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with water (2 x 100 mL) and brine (100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1), to give the title compound as a yellow solid. The racemic mixture was purified by SFC under the following conditions (column: NB_ASACHIRALPAK IG_2, 5*30 cm, 10 μm; mobile phase A: CO2, mobile phase B: MeOH--HPLC; flow rate: 200 mL / min; gradient: isocratic 55% B; column temperature (°C): 35; back pressure (bar): 100; wavelength: 290 nm; RT1 (min): 4.04; RT2 (min): 6.28; sample solvent: MeOH:DCM=1:2; injection volume: 6.5 mL; number of runs: 26), yielding enantiomer 1 (6.5 g) and enantiomer 2 (8.0 g), which were pale yellow solids.

[0523] Enantiomer 1 (Example 173) was further purified by grinding with ACN (20 mL) to give the title compound (5.6 g, 20%) as a grayish-white solid. LCMS (ES, m / z): [M+H]+ =385. 1H NMR (400 MHz, DMSO-d6) δ13.06 (s, 1H), 7.89 – 7.79 (m, 2H), 7.33 – 7.22 (m, 2H), 4.50 (dd, J = 11.1,2.3 Hz, 1H), 3.99 (m, 3H), 3.57 (m, 1H), 3.27 (t, J = 2.6 Hz, 1H), 2.12 –2.01 (m, 1H), 1.90 (m, 1H), 1.73 (br d, J = 13.2 Hz, 1H), 1.54-1.64 (m, 3H).

[0524] Enantiomer 2 (Example 174) was purified by reversed-phase rapid chromatography under the following conditions (column, C18 silica gel; mobile phase, aqueous ACN solution (0.1% FA), gradient 50% to 70% over 5 min; detector, UV 254 nm), concentrated under reduced pressure, and further purified by grinding with ACN (12 mL) to give the title compound (5.65 g, 20%) as a grayish-white solid. LCMS (ES,m / z): [M+H]+ =385. 1H NMR (400 MHz, DMSO-d6) δ 13.06 (s, 1H), 7.89 – 7.79 (m,2H), 7.33 – 7.22 (m, 2H), 4.50 (dd, J = 11.1, 2.3 Hz, 1H), 3.99 (m, 3H), 3.57(m, 1H), 3.27 (t, J = 2.6 Hz, 1H), 2.12 – 2.01 (m, 1H), 1.90 (m, 1H), 1.73(br d, J = 13.2 Hz, 1H), 1.54-1.64 (m, 3H).

[0525] Example 175-7-Methyl-8-{4-[(1-methylpiperidin-4-yl)oxy]phenyl}-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0526]

[0527] Step 1: 8-(4-hydroxyphenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazine-4-one

[0528] At room temperature, K₂CO₃ (2.48 g, 17.9 mmol) and Pd(Dtbpf)Cl₂ (0.39 g, 0.60 mmol, CAS: 95408-45-0) were added to a stirred mixture of 8-bromo-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazine-4-one (3.0 g, 6.0 mmol) and 4-hydroxyphenylboronic acid (0.99 g, 7.2 mmol) in dioxane (45 mL) and water (15 mL). The resulting mixture was stirred overnight at 80 °C under N₂. The residue was dissolved in water (60 mL). The resulting mixture was extracted with EA (3 x 60 mL). The combined organic layers were washed with water (2 x 60 mL) and brine (60 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1) to give 8-(4-hydroxyphenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one (1.68 g, 55%), as a brown solid. LCMS 229-1: (ES, m / z): [M+H] + =515.

[0529] Step 2: 3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methyl-8-{4-[(1-methylpiperidin-4-yl)oxy]phenyl}pyrazolo[1,5-a][1,3,5]triazin-4-one

[0530] At room temperature, PPh3 (816 mg, 3.11 mmol) was added to a stirred solution of 8-(4-hydroxyphenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one (800 mg, 1.56 mmol) and 1-methyl-4-piperidinol (1.79 g, 15.6 mmol) in THF (16 mL). DIAD (629 mg, 3.11 mmol) was added dropwise to the mixture at 0 °C under N2. The resulting mixture was stirred again overnight at room temperature under N2. The resulting mixture was concentrated under vacuum. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN aqueous solution (0.1% NH3·H2O), 30% to 60% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (915 mg, 96%), as a yellow solid. LCMS (ES, m / z): [M+H] + =612.

[0531] Step 3: 7-Methyl-8-{4-[(1-methylpiperidin-4-yl)oxy]phenyl}-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0532] A solution of 3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methyl-8-{4-[(1-methylpiperidin-4-yl)oxy]phenyl}pyrazolo[1,5-a][1,3,5]triazin-4-one (915 mg, 1.50 mmol) in TFA (5 mL) was stirred for 2 h at 70 °C. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution (0.1% FA), 30% to 80% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (547 mg, 98%) as a brown solid. LCMS (ES, m / z): [M+H] + =372.

[0533] Step 4: 7-Methyl-8-{4-[(1-methylpiperidin-4-yl)oxy]phenyl}-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0534] At room temperature, K₂CO₃ (611 mg, 4.42 mmol) and propargyl bromide (88 mg, 0.74 mmol) were added to a stirred solution of 7-methyl-8-{4-[(1-methylpiperidin-4-yl)oxy]phenyl}-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (547 mg, 1.47 mmol) in DMF (10 mL). The resulting mixture was stirred at room temperature for 1 h. The mixture was filtered, and the filter cake was washed with DMF (3 x 1 mL). The filtrate was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 20% to 50% gradient over 10 min; detector, UV 254 nm, to give the crude product. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 12% B to 29% B, 29% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 4.05), yielding the title compound (41 mg, 7%) as a white solid. LCMS (ES, m / z): [M+H] + =410. 1H NMR (400 MHz, DMSO-d6) δ 7.67 – 7.65 (m, 2H), 7.04 – 7.02 (m,2H), 4.61 (m, 1H), 3.89 (d, J = 2.7 Hz, 2H), 3.27 (m, 2H), 3.10 – 3.09 (m,3H), 2.75 (s, 3H), 2.39 (s, 3H), 2.08 (t, J = 11.7 Hz, 2H), 1.92 (t, J = 11.7Hz, 2H).

[0535] Synthesis method M

[0536] Example 176-8-(4-chlorophenyl)-7-isopropyl-2-(prop-2-yn-1-yloxy)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0537]

[0538] Step 1: 2-(4-chlorophenyl)-4-methyl-3-oxopentanilide

[0539] NaH (1.24 g, 51.65 mmol, 60% in mineral oil) was added to a stirred solution of 4-chlorobenzyl cyanide (2.74 g, 18.1 mmol) in THF (30 mL). The resulting mixture was stirred at 0 °C under N2 for 30 min. Ethyl isobutyrate (3.00 g, 25.8 mmol) was added to the mixture at 0 °C. The resulting mixture was stirred again overnight at room temperature. The mixture was acidified to pH 6 with 2 M HCl (aqueous solution), diluted with water (20 mL), and extracted with EA (3 x 30 mL). The combined organic layers were washed with brine (2 x 30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (9:1) to give the title compound (3.66 g, 64%) as a yellow liquid. LCMS (ES, m / z): [MH] - =220.

[0540] Step 2: 3-Nitro-4-Phenyridine

[0541] At room temperature, hydrazine hydrate (2.01 g, 32.1 mmol, 80 wt%) and TFA (3.66 g, 32.1 mmol) were added to a stirred solution of 2-(4-chlorophenyl)-4-methyl-3-oxopentanonitrile (3.56 g, 16.1 mmol) in EtOH (35 mL). The resulting mixture was stirred at 80 °C for 2 h and concentrated under reduced pressure. The residue was dissolved in THF (10 mL) and purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution (0.1% NH3·H2O) in a 30% to 60% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (3.57 g, 94%) as a yellow oil. LCMS (ES, m / z): [M+H] + =236.

[0542] Step 3: N-{[4-(4-chlorophenyl)-5-isopropyl-2H-pyrazol-3-yl]aminothioyl}carbamate

[0543] At room temperature, ethyl N-thiocarbonylcarbamate (1.06 g, 8.06 mmol) was added to a stirred solution of 4-(4-chlorophenyl)-5-isopropyl-2H-pyrazole-3-amine (1.9 g, 8.1 mmol) in DMF (19 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was then poured into ice / water (40 mL). The precipitated solid was collected by filtration, washed with water (3 x 5 mL), and dried under vacuum. This yielded the title compound (2.5 g, 85%) as a yellow solid. LCMS (ES, m / z): [M+H] + =367.

[0544] Step 4: 3-Nitro-4-Phenyridine

[0545] EtONa (891 mg, 13.1 mmol) was added to a stirred solution of N-{[4-(4-chlorophenyl)-5-isopropyl-2H-pyrazol-3-yl]aminothioyl}carbamate (2.4 g, 6.5 mmol) in EtOH (24 mL). The resulting mixture was stirred at 80 °C for 2 h and concentrated under reduced pressure. The residue was dissolved in water (20 mL) and acidified to pH 6 with 2 M HCl (aqueous solution). The precipitated solid was collected by filtration, washed with water (3 x 5 mL), and dried under vacuum. This yielded the title compound (2.0 g, 95%) as a yellow solid. LCMS (ES, m / z): [M+H] + =321.

[0546] Step 5: 8-(4-chlorophenyl)-7-isopropyl-2-(methylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0547] At room temperature, K₂CO₃ (2.71 g, 19.6 mmol) and MeI (1.02 g, 7.20 mmol) were added to a stirred solution of 8-(4-chlorophenyl)-7-isopropyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (2.1 g, 6.55 mmol) in DMF (21 mL). The resulting mixture was stirred at room temperature for 1 h. The residue was dissolved in water (20 mL). The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with water (2 x 20 mL) and brine (2 x 20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (1:2), to give the title compound (900 mg, 41.06%) as a grayish-white solid. LCMS (ES, m / z): [M+H]+ =335.

[0548] Step 6: 8-(4-chlorophenyl)-7-isopropyl-2-methanesulfonyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one and 8-(4-chlorophenyl)-7-isopropyl-2-(methylsulfinyl)pyrazolo[1,5-a][1,3,5]triazin-4(3H)-one

[0549] At 0 °C, mCPBA (1020 mg, 5.91 mmol) was added fractionally to a stirred solution of 8-(4-chlorophenyl)-7-isopropyl-2-(methylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (990 mg, 2.96 mmol) in DCM (20 mL). The resulting mixture was stirred at 0 °C for 15 min and then at room temperature for 2 h, followed by concentration under reduced pressure. The residue was dissolved in THF (10 mL). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution (0.1% FA), 30% to 80% gradient over 10 min; detector, UV 254 nm. This yielded a mixture of the title compound (880 mg, 81%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =351 and 367.

[0550] Step 7: 8-(4-chlorophenyl)-7-isopropyl-2-(prop-2-yn-1-yloxy)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0551] At 0 °C, NaH (28 mg, 0.71 mmol, 60% in mineral oil) was added in portions to a stirred solution of propargyl alcohol (60 mg, 1.06 mmol) in DMF (2.6 mL). The resulting mixture was stirred at 0 °C for 30 min. At room temperature, a mixture of 8-(4-chlorophenyl)-7-isopropyl-2-methanesulfonyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one and 8-(4-chlorophenyl)-7-isopropyl-2-(methylsulfinyl)pyrazolo[1,5-a][1,3,5]triazine-4(3H)-one (130 mg, 0.354 mmol) was added to the above mixture. The resulting mixture was stirred at room temperature for another 2 h. The mixture was acidified to pH 7 with 1 M HCl (aqueous solution). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN (0.1% FA), gradient from 30% to 80% over 10 min; detector, UV 254 nm, yielding the crude product. The crude product (70 mg) was purified by Prep-HPLC under the following conditions (column: Sunfire prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 45% B to 70% B, 70% B over 8 min; wavelength: 254 / 220 nm; RT1 (min): 7.05), yielding the title compound (53 mg, 43%) as a white solid. LCMS (ES, m / z): [M+H] + = 343. 1 H NMR (400 MHz, DMSO-d6) δ 12.85 (s, 1H), 7.60 – 7.58 (m,2H), 7.51 – 7.49 (m, 2H), 5.03 – 5.02 (d, J = 2.5 Hz, 2H), 3.73 – 3.71 (t, J= 2.4 Hz, 1H), 3.33 – 3.24 (m, 1H), 1.24 – 1.16 (d, J = 6.8 Hz, 6H).

[0552] The examples in Table M were synthesized using synthesis method M:

[0553] Table M

[0554]

[0555]

[0556]

[0557]

[0558]

[0559]

[0560]

[0561]

[0562]

[0563]

[0564]

[0565]

[0566]

[0567]

[0568]

[0569]

[0570] Example 239-7-(difluoromethyl)-2-(prop-2-yn-1-yloxy)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0571]

[0572] Step 1: 4,4-Difluoro-3-oxo-2-(3,4,5-trifluorophenyl)butyronitrile

[0573] At 0 °C, NaH (3.51 g, 87.7 mmol, 60% in mineral oil) was added fractionally to a stirred solution of 2-(3,4,5-trifluorophenyl)acetonitrile (5.0 g, 29 mmol) in THF (50 mL). The resulting mixture was stirred at 0 °C for 30 min. At 0 °C, ethyl 2,2-difluoroacetate (10.9 g, 87.7 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for another 4 h. The mixture was acidified to pH 5 with 2N HCl (2 mL). The resulting mixture was diluted with water (100 mL). The resulting mixture was extracted with EA (3 x 150 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (5:1) to give the title compound (4.9 g, 67%) as a pink solid. LCMS (ES, m / z): [MH] - =248.

[0574] Step 2: 5-(difluoromethyl)-4-(3,4,5-trifluorophenyl)-2H-pyrazole-3-amine

[0575] At room temperature, hydrazine hydrate (3.01 g, 48.2 mmol, 80 wt%) and TFA (8.24 g, 72.2 mmol) were added to a stirred solution of 4,4-difluoro-3-oxo-2-(3,4,5-trifluorophenyl)butyronitrile (3.0 g, 12 mmol) in EtOH (30 mL). The resulting mixture was stirred at 60 °C for 3 h. The mixture was concentrated under reduced pressure. Water (100 mL) was added to the mixture. The resulting mixture was extracted with EA (3 x 120 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (3.4 g, overweight), a yellow oil that could be used directly in the next step without further purification. LCMS (ES, m / z): [M+H]⁺ = 264.

[0576] Step 3: N-{[5-(difluoromethyl)-4-(3,4,5-trifluorophenyl)-2H-pyrazol-3-yl]aminothioyl}carbamate

[0577] At room temperature, ethyl N-thiocarbonylcarbamate (1.69 g, 12.9 mmol) was added to a stirred solution of 5-(difluoromethyl)-4-(3,4,5-trifluorophenyl)-2H-pyrazole-3-amine (12 mmol) in DMF (34 mL). The resulting mixture was stirred at room temperature for 1 h. Water (400 mL) was added to the mixture. The precipitated solid was collected by filtration and washed with water (2 x 20 mL). This yielded the title compound (5.0 g, overweight), a grayish-white solid, which could be used directly in the next step without further purification. LCMS (ES, m / z): [MH] - =393.

[0578] Step 4: 7-(difluoromethyl)-2-thioalkyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0579] A solution of N-{[5-(difluoromethyl)-4-(3,4,5-trifluorophenyl)-2H-pyrazol-3-yl]carbamate ethyl}carbamate (12 mmol) in 2 N NaOH (100 mL) was stirred at room temperature for 1 h. The mixture was acidified to pH 6 with 2 N HCl (25 mL). The precipitated solid was collected by filtration and washed with water (2 x 10 mL). This yielded the title compound (3.0 g, 72% (step 3)) as a grayish-white solid, which could be used directly for the next step without further purification. LCMS (ES, m / z): [MH] - =347.

[0580] Step 5: 7-(difluoromethyl)-2-(methylthioalkyl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0581] At room temperature, K₂CO₃ (560 mg, 4.05 mmol) and methyl iodine (172 mg, 1.22 mmol) were added to a stirred solution of 7-(difluoromethyl)-2-thioalkyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (470 mg, 1.35 mmol) in DMF (5 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was diluted with water (30 mL). The resulting mixture was extracted with EA (3 x 30 mL). The combined organic layers were washed with water (20 mL) and brine (2 x 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 55% to 75% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (480 mg, 98%), a white solid. LCMS (ES, m / z): [M+H] + =363.

[0582] Step 6: 7-(difluoromethyl)-2-methanesulfonyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0583] At 0 °C, mCPBA (457 mg, 2.65 mmol) was added to a stirred solution of 7-(difluoromethyl)-2-(methylthioalkyl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (480 mg, 1.33 mmol) in DCM (5 mL). The resulting mixture was stirred at 0 °C for 15 min and then at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 50% to 70% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (400 mg, 77%) as a white solid. LCMS (ES, m / z): [MH] - =393.

[0584] Step 7: 7-(difluoromethyl)-2-(prop-2-yn-1-yloxy)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0585] At 0 °C, NaH (61 mg, 1.5 mmol, 60% mineral oil) was added to a stirred solution of propargyl alcohol (43 mg, 0.76 mmol) in DMF (4 mL). The resulting mixture was stirred at 0 °C for 30 min. At 0 °C, 7-(difluoromethyl)-2-methanesulfonyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (200 mg, 0.507 mmol) was added to the above mixture. The resulting mixture was stirred at room temperature for another 2 h. The mixture was acidified to pH 5 with 2N HCl (0.4 mL). The resulting mixture was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 70% to 90% gradient over 10 min; detector, UV 254 nm. The crude product (180 mg) was purified by Prep-HPLC under the following conditions (column: Xselect CSH C18 OBD column 30*150 mm 5 μm, n; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 43% B to 58% B over 9 min; wavelength: 254 / 220 nm; RT1 (min): 7.80), yielding the title compound (87.6 mg, 47%) as a white solid. LCMS (ES, m / z): [M+H] + =371. 1H NMR (400 MHz, DMSO-d6) δ 13.37 (s, 1H), 7.64 - 7.59 (m, 2H), 7.35 (t, J = 53.0 Hz, 1H), 5.12 (d, J = 2.4 Hz, 2H), 3.78 (t, J = 2.4 Hz, 1H), 2.09 (s, 1H).

[0586] Example 240-2-(2,2-difluoroethoxy)-7-(difluoromethyl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0587]

[0588] At 0 °C, NaH (30 mg, 0.76 mmol, 60% in mineral oil) was added to a stirred solution of 2,2-difluoroethanol (31 mg, 0.381 mmol) in DMF (2 mL). The resulting mixture was stirred at 0 °C for 30 min. At 0 °C, 7-(difluoromethyl)-2-methanesulfonyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (100 mg, 0.254 mmol) was added to the above mixture. The resulting mixture was stirred at room temperature for another 2 h. The mixture was acidified to pH 5 with 2N HCl (0.4 mL). The resulting mixture was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 70% to 90% gradient over 10 min; detector, UV 254 nm. The crude product (100 mg) was purified by Prep-HPLC under the following conditions (column: Sunfire prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.05% TFA), mobile phase B: MeOH--HPLC; flow rate: 60 mL / min; gradient: 63% 83% B over 10 min; wavelength: 254 nm / 220 nm; RT1 (min): 10.07), yielding the title compound (67.3 mg, 67%) as a white solid. LCMS (ES, m / z): [M+H]+=397. 1H NMR (400 MHz, DMSO-d6) δ 13.57 (s, 1H), 7.57 – 7.53 (m, 2H), 7.35 (t,J = 53.0 Hz, 1H), 6.45 (tt, J = 53.7, 3.0 Hz, 1H), 4.74 (td, J = 15.2, 3.0Hz, 2H).

[0589] Example 241 - 8-{4-[2-(dimethylamino)ethoxy]phenyl}-7-methoxy-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0590]

[0591] Step 1: 2-(5-hydroxy-2H-pyrazol-3-yl)isoindole-1,3-dione

[0592] Phthalic anhydride (17.9 g, 121 mmol) was added to a stirred solution of 5-amino-1H-pyrazole-3-ol (10.0 g, 101 mmol) in acetic acid (100 mL) at room temperature under N2. The resulting mixture was stirred at 130 °C for 2 h. The mixture was then cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by grinding with EA (50 mL). The resulting solid was dried under vacuum to give the title compound (20 g, 87%) as an orange solid. LCMS (ES, m / z): [M+H]+ = 230.

[0593] Step 2: 2-(5-methoxy-2H-pyrazol-3-yl)isoindole-1,3-dione

[0594] PPh3 (34.3 g, 131 mmol) was added to a stirred mixture of 2-(5-hydroxy-2H-pyrazol-3-yl)isoindole-1,3-dione (20 g, 87 mmol) and methanol (20 mL) in THF (200 mL) at 0 °C under N2. The resulting mixture was stirred at 0 °C for 5 min. DIAD (51.9 mL, 262 mmol) was added to the mixture at 0 °C. The resulting mixture was stirred again at 60 °C overnight. The mixture was cooled to room temperature. The resulting mixture was diluted with water (200 mL). The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1) to give the title compound (10 g, 48%) as an orange solid. LCMS (ES, m / z): [M+H]+ =244.

[0595] Step 3: 2-(4-bromo-5-methoxy-2H-pyrazol-3-yl)isoindole-1,3-dione

[0596] NBS (2.93 g, 16.4 mmol) was added to a stirred solution of 2-(5-methoxy-2H-pyrazol-3-yl)isoindole-1,3-dione (4.0 g, 16 mmol) in ACN (40 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1), to give the two title compounds (3.0 g, 57%) as a brown solid. LCMS (ES, m / z): [M+H] + =322.

[0597] Step 4: 4-Bromo-5-methoxy-2H-pyrazole-3-amine

[0598] At room temperature, hydrazine hydrate (80%) (2.91 g, 46.6 mmol) was added to a stirred solution of 2-(4-bromo-5-methoxy-2H-pyrazol-3-yl)isoindole-1,3-dione (3.0 g, 9.3 mmol) in EtOH (30 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The mixture was diluted with water (50 mL). The mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with brine (2 x 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% NH₄) 3- H₂O), gradient of 10% to 50% over 10 min; detector, UV 254 nm, yielded the title compound (500 mg, 28%) as a yellow solid. LCMS (ES, m / z): [M+H] + =192.

[0599] Step 5: N-[(4-bromo-5-methoxy-2H-pyrazol-3-yl)aminothioyl]carbamate

[0600] At room temperature, ethyl N-thiocarbonylcarbamate (376 mg, 2.86 mmol) was added to a stirred solution of 4-bromo-5-methoxy-2H-pyrazole-3-amine (500 mg, 2.60 mmol) in DMF (10 mL). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give the title compound (640 mg, 76%) as an orange solid. LCMS (ES, m / z): [M+H]+=323.

[0601] Step 6: 8-Bromo-7-methoxy-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0602] EtONa (270 mg, 3.96 mmol) was added to a stirred solution of N-[(4-bromo-5-methoxy-2H-pyrazol-3-yl)aminothioyl]carbamate (640 mg, 1.98 mmol) in EtOH (12 mL) at 0 °C. The resulting mixture was stirred at 80 °C for 1 h. The mixture was then cooled to room temperature. The resulting mixture was diluted with water (10 mL). The mixture was acidified to pH 5 with 2 M HCl. The precipitated solid was collected by filtration and washed with water (3 x 1 mL). The resulting solid was dried under vacuum to give the title compound (450 mg, 82%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =277.

[0603] Step 7: 8-Bromo-7-methoxy-2-{[(4-methoxyphenyl)methyl]thioalkyl}-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0604] At room temperature, PMBCl (763 mg, 4.87 mmol) was added to a stirred mixture of 8-bromo-7-methoxy-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (450 mg, 1.62 mmol) and K₂CO₃ (673 mg, 4.87 mmol) in DMF (10 mL). The resulting mixture was stirred overnight at room temperature. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (2 x 10 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (520 mg, 81%) as a brown solid. LCMS(ES, m / z) :[M+H] - =395.

[0605] Step 8: 8-Bromo-7-methoxy-3-(methoxymethyl)-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazin-4-one

[0606] At 0 °C, NaH (60% in mineral oil, 63 mg, 2.6 mmol) was added in portions to a stirred solution of 8-bromo-7-methoxy-2-{[(4-methoxyphenyl)methyl]thioalkyl}-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (520 mg, 1.31 mmol) in DMF (10 mL). The resulting mixture was stirred at 0 °C for 20 min. At 0 °C, bromomethyl methyl ether (649 mg, 5.24 mmol) was added to the mixture. The resulting mixture was stirred again overnight at room temperature. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (2 x 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 10% to 100% gradient over 10 min; detector, UV 254 nm, to give the title compound (300 mg, 52%) as a yellow solid. LCMS (ES, m / z): [M+H] + =441.

[0607] Step 9: Dimethyl({2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)phenoxy]ethyl})amine

[0608] DIAD (1.38 g, 6.82 mmol) was added to a stirred mixture of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)phenol (1.0 g, 4.5 mmol), dimethylaminoethanol (0.61 g, 6.8 mmol), and PPh3 (1.79 g, 6.82 mmol) in THF (20 mL) at 0 °C under N2. The resulting mixture was stirred overnight at room temperature under N2. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1), to give the title compound (600 mg, 45%) as a white solid. LCMS (ES,m / z): [M+H] + =292.

[0609] Step 10: 8-{4-[2-(dimethylamino)ethoxy]phenyl}-7-methoxy-3-(methoxymethyl)-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazine-4-one

[0610] At room temperature under N2, KHCO3 (136 mg, 1.36 mmol) and Pd(dtbpf)Cl2 (67 mg, 0.10 mmol) were added to a stirred mixture of 8-bromo-7-methoxy-3-(methoxymethyl)-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazine-4-one (300 mg, 0.680 mmol) and dimethyl({2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)phenoxy]ethyl})amine (396 mg, 1.36 mmol) in dioxane (6 mL) and water (0.6 mL). The resulting mixture was stirred overnight at 80 °C under N2. The mixture was then cooled to room temperature. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 10 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (110 mg, 20%) as a white solid. LCMS (ES, m / z): [M+H] + =526.

[0611] Step 11: 8-{4-[2-(dimethylamino)ethoxy]phenyl}-7-methoxy-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0612] A solution of 8-{4-[2-(dimethylamino)ethoxy]phenyl}-7-methoxy-3-(methoxymethyl)-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazin-4-one (110 mg, 0.209 mmol) in TFA (2 mL) was stirred for 1 h at 70 °C under N2. The mixture was cooled to room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (20 mg, 26%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =362.

[0613] Step 12: 8-{4-[2-(dimethylamino)ethoxy]phenyl}-7-methoxy-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0614] At room temperature under N2, propargyl bromide (4.0 mg, 0.033 mmol) was added dropwise to a stirred mixture of 8-{4-[2-(dimethylamino)ethoxy]phenyl}-7-methoxy-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (20 mg, 0.055 mmol) and K2CO3 (23 mg, 0.17 mmol) in DMF (1 mL). The resulting mixture was stirred at room temperature under N2 for 1 h. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 5 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 8% B to 35% B, 35% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 4.40), yielding the title compound (1.1 mg, 5%) as a white solid. LCMS (ES, m / z): [M+H] + =400. 1H NMR (300 MHz, DMSO-d6) 8.02 (d, J = 8.2 Hz, 2H), 6.95 (d, J = 8.3 Hz, 2H), 4.18 (t, 2H), 4.06 (s, 3H), 3.96 (s, 2H), 3.09 (m, 3H), 2.53 (s, 6H).

[0615] Example 242-8-(4-fluorophenyl)-7-(1H-imidazol-4-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0616]

[0617] Step 1: 1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-carboxylic acid ethyl ester

[0618] At 0 °C, NaH (285 mg, 7.14 mmol, 60% in mineral oil) was added fractionally to a stirred solution of ethyl 1H-imidazolium-4-carboxylate (500 mg, 3.57 mmol) in DMF (10 mL). The resulting mixture was stirred at 0 °C for 1 h. SEMCl (1.78 g, 10.7 mmol) was added to the mixture at room temperature. The resulting mixture was stirred at room temperature for another 2 h. The reaction was quenched with water / ice (20 mL) at 0 °C. The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with water (2 x 20 mL) and brine (20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN aqueous solution (0.1% FA), in a 30% to 80% gradient over 10 min; detector, UV 254 nm, yielding the title compound (705 mg, 73%) as a red oil. LCMS (ES, m / z): [M+H] + =271.

[0619] Step 2: 2-(4-fluorophenyl)-3-oxo-3-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)propionitrile

[0620] At 0 °C, NaH (178 mg, 4.44 mmol, 60% in mineral oil) was added in portions to a stirred solution of 4-fluorophenylacetonitrile (240 mg, 1.78 mmol) in THF (12 mL). The resulting mixture was stirred at 0 °C for 30 min. Ethyl 1-{[2-(trimethylsilyl)ethoxy]methyl}imidazolium-4-carboxylate (600 mg, 2.22 mmol) was added to the mixture at room temperature. The resulting mixture was stirred again overnight at room temperature. The reaction was quenched by adding 20 mL of water at 0 °C. The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (2 x 20 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN (0.1% FA), 40% to 90% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (320 mg, 40%) as a yellow solid. LCMS (ES, m / z): [M+H] + =360.

[0621] Step 3: 4-(4-fluorophenyl)-5-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)-2H-pyrazole-3-amine

[0622] At room temperature, hydrazine hydrate (75 mg, 0.22 mmol, 80 wt%) and TFA (171 mg, 1.50 mmol) were added to a stirred solution of 2-(4-fluorophenyl)-3-oxo-3-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)propionitrile (270 mg, 0.751 mmol) in EtOH (5.4 mL). The resulting mixture was stirred overnight at 40 °C. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN (0.1% FA), 30% to 80% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (211 mg, 75%) as a brown solid. LCMS (ES, m / z): [M+H] + =374.

[0623] Step 4: N-{[4-(4-fluorophenyl)-5-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)-2H-pyrazol-3-yl]carbamate}ethyl carbamate

[0624] At room temperature, N-thiocarbonylcarbamate (56 mg, 0.43 mmol) was added to a stirred solution of 4-(4-fluorophenyl)-5-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)-2H-pyrazole-3-amine (160 mg, 0.428 mmol) in DMF (3.2 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 15 mL). The combined organic layers were washed with water (2 x 15 mL) and brine (2 x 15 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (160 mg, 74%) as a brown oil. The crude product was used directly for the next step without further purification. LCMS (ES, m / z): [M+H] + = 505.

[0625] Step 5: 8-(4-fluorophenyl)-2-thioalkyl-7-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0626] A solution of N-{[4-(4-fluorophenyl)-5-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)-2H-pyrazol-3-yl]carbamate (160 mg, 0.317 mmol) in 2M NaOH (3 mL) was stirred at room temperature for 30 min. The residue was acidified to pH 6 with 2M HCl (2 mL) and purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 30% to 80% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (105 mg, 72%) as a brown solid. LCMS (ES, m / z): [M+H] + =459.

[0627] Step 6: 8-(4-fluorophenyl)-2-(prop-2-yn-1-ylthioalkyl)-7-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0628] At room temperature, K₂CO₃ (95 mg, 0.69 mmol) and propargyl bromide (24 mg, 0.21 mmol) were added to a stirred solution of 8-(4-fluorophenyl)-2-thioalkyl-7-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (105 mg, 0.229 mmol) in DMF (2.8 mL). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was filtered, and the filter cake was washed with DMF (3 x 1 mL). The filtrate was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution (0.1% FA), 30% to 80% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (84 mg, 74%) as a brown solid. LCMS (ES,m / z): [M+H] + =497.

[0629] Step 7: 8-(4-fluorophenyl)-7-(1H-imidazol-4-yl)-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0630] A solution of 8-(4-fluorophenyl)-2-(prop-2-yn-1-ylthioalkyl)-7-(1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (64 mg, 0.13 mmol) in HCl (4 M solution in 1,4-dioxane, 4.5 mL) was stirred overnight at room temperature. The reaction was quenched with water at 0 °C. The resulting mixture was purified by reverse-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN solution (0.1% FA), 10% to 50% gradient over 10 min; detector, UV 254 nm. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 15% B to 33% B, 33% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 5.40), yielding the title compound (14 mg, 30%) as a white solid. LCMS (ES, m / z): [M+H] + =367. 1 H NMR (400 MHz, DMSO-d6) δ 13.21 (s, 1H), 8.05 (s, 1H), 7.67 – 7.63 (m, 2H), 7.27 (s, 1H), 7.22 –7.17 (m, 2H), 3.93 (d, J = 2.6 Hz, 2H), 3.19 (t, J = 2.7 Hz, 1H).

[0631] Example 243-2-(2,2-difluoroethoxy)-7-isopropyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0632]

[0633] Step 1: 4-Methyl-3-oxo-2-(3,4,5-trifluorophenyl)pentanilonitrile

[0634] A solution of 2-(3,4,5-trifluorophenyl)acetonitrile (1.00 g, 5.84 mmol) in THF (20 mL) was treated with NaH (0.28 g, 12 mmol, 60% in mineral oil) at 0 °C for 30 min, followed by dropwise addition of isobutyryl chloride (1.87 g, 17.5 mmol) at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by adding 20 mL of water at 0 °C. The resulting mixture was extracted with EA (3 x 30 mL). The combined organic layers were washed with water (30 mL) and dried over anhydrous Na₂SO₄. The residue was purified by silica gel column chromatography, eluting with PE / EA (7:1), to give the title compound (1.00 g, 71%) as a yellow solid. LCMS (ES, m / z): [MH] - =240.

[0635] Step 2: 5-Isopropyl-4-(3,4,5-trifluorophenyl)-2H-pyrazole-3-amine

[0636] A solution of 4-methyl-3-oxo-2-(3,4,5-trifluorophenyl)pentanonitrile (1.10 g, 4.56 mmol), hydrazine hydrate (0.46 g, 9.12 mmol), and TFA (0.7 mL, 9.12 mmol) in EtOH (20 mL) was stirred for 2 h at 65 °C. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (1.20 g, overweight), as a yellow oil. LCMS (ES, m / z): [M+H] + =257.

[0637] Step 3: N-{[5-isopropyl-4-(3,4,5-trifluorophenyl)-2H-pyrazol-3-yl]carbamoyl}ethyl carbamate

[0638] A solution of 5-isopropyl-4-(3,4,5-trifluorophenyl)-2H-pyrazole-3-amine (4.56 mmol) and N-thiocarbonylcarbamate (0.45 g, 3.45 mmol) in DMF (20 mL) was stirred for 2 h at room temperature. The reaction was quenched by adding water / ice (50 mL) at 0 °C. The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with water (50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (1.50 g, 85%) as a yellow oil that solidified upon standing. LCMS (ES, m / z): [M+H] + =387

[0639] Step 4: 7-Isopropyl-2-thioalkyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0640] A solution of N-{[5-isopropyl-4-(3,4,5-trifluorophenyl)-2H-pyrazol-3-yl]carbamate (4.50 g, 11.7 mmol) in NaOH (2 M aqueous solution, 50 mL) was stirred at room temperature for 2 h. The mixture was acidified to pH 6 with concentrated HCl. The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (1.90 g, 48%) as a yellow solid. LCMS (ES, m / z): [M+H] + =341.

[0641] Step 5: 7-Isopropyl-2-(methylthioalkyl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0642] A solution of 7-isopropyl-2-thioalkyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (400 mg, 1.17 mmol), MeI (100 mg, 0.70 mmol), and K₂CO₃ (487 mg, 3.52 mmol) in DMF (4 mL) was stirred for 2 h at room temperature. The reaction was quenched with water (10 mL) at room temperature. The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (200 mg, 48%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =355.

[0643] Step 6: 7-Isopropyl-2-methanesulfonyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one and 7-isopropyl-2-methanesulfinyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0644] A solution of 7-isopropyl-2-(methylthioalkyl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (300 mg, 0.85 mmol) and mCPBA (292 mg, 1.69 mmol) in DCM (6 mL) was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in MeOH (5 mL). The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 10% to 50% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (150 mg, 46%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =371 and 387.

[0645] Step 7: 2-(2,2-difluoroethoxy)-7-isopropyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0646] At 0 °C, NaH (9.0 mg, 0.39 mmol, 60% mineral oil) was added in portions to a stirred solution of 2,2-difluoroethanol (32 mg, 0.39 mmol) in DMF (1 mL). The resulting mixture was stirred at 0 °C for 30 min. At room temperature, a mixture of 7-isopropyl-2-methanesulfonyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one and 7-isopropyl-2-methanesulfinyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (50 mg, 0.13 mmol, Mw: 386) was added. The resulting mixture was stirred at room temperature for 1 h. The mixture was acidified to pH 7 with 1 M HCl (aqueous solution). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN (0.1% FA), gradient from 30% to 80% over 10 min; detector, UV 254 nm, yielding the crude product. The crude product was then purified by Prep-HPLC under the following conditions (column: Xselect CSH C18 OBD column 30*150 mm 5 μm, n; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 45% B to 57% B, 57% B over 10 min; wavelength: 254 / 220 nm; RT1 (min): 9.28), yielding the title compound (25 mg, 50%) as a white solid. LCMS (ES, m / z): [M+H] + =389. 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 13.07 (s, 1H), 7.49 – 7.45 (m, 2H), 6.42 (tt, J = 53.8, 3.1 Hz, 1H), 4.66 (td, J = 15.1, 3.1 Hz, 2H), 3.32 (septet, J = 6.8 Hz, 1H), 1.22 (d, J = 6.8 Hz, 6H).

[0647] Example 244-7-Isopropyl-2-(prop-2-yn-1-yloxy)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0648]

[0649] At 0 °C, NaH (13 mg, 0.54 mmol, 60% in mineral oil) was added in portions to a stirred solution of propargyl alcohol (30 mg, 0.54 mmol) in DMF (1 mL). The resulting mixture was stirred at 0 °C for 30 min. At room temperature, a mixture of 7-isopropyl-2-methanesulfonyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one and 7-isopropyl-2-methanesulfinyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (70 mg, 0.18 mmol, Mw: 386) was added. The resulting mixture was stirred at room temperature for 1 h. The mixture was acidified to pH 7 with 1 M HCl (aqueous solution). The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN (0.1% FA), gradient from 30% to 80% over 10 min; detector, UV 254 nm, yielding the crude product. The crude product (100 mg) was purified by Prep-HPLC under the following conditions (column: Xselect CSH C18 OBD column 30*150 mm 5 μm, n; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 44% B to 57% B, 57% B over 10 min; wavelength: 254 / 220 nm; RT1 (min): 9.02), yielding the title compound (20 mg, 30%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =363. 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 12.96 (s, 1H), 7.52 – 7.48 (m, 2H), 5.05 (d, J = 2.5 Hz, 2H), 3.73 (t, J = 2.5 Hz, 1H), 3.34 (septet, J = 6.7 Hz, 1H), 1.21 (d, J = 6.8 Hz, 6H).

[0650] Synthesis method Q

[0651] Example 245-2-(2,2-difluoroethoxy)-7-methyl-8-[4-(pyrrolidine-1-carbonyl)phenyl]-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0652]

[0653] Step 1: 3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methyl-8-[4-(pyrrolidine-1-carbonyl)phenyl]pyrazolo[1,5-a][1,3,5]triazine-4-one

[0654] To a mixture of 8-bromo-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methylpyrazolo[1,5-a][1,3,5]triazin-4-one (1.0 g, 2.0 mmol) and 4-(pyrrolidine-1-carbonyl)phenylboronic acid (655 mg, 2.99 mmol) in dioxane (8 mL) and water (2 mL), K₂CO₃ (827 mg, 5.98 mmol) and Pd(dtbpf)Cl₂ (130 mg, 0.199 mmol) were added. The resulting mixture was stirred overnight at 80 °C. Water (20 mL) was added to the mixture. The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with water (10 mL) and brine (10 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN aqueous solution (0.1% FA), in a 10% to 80% gradient over 10 min; detector, UV 254 nm, yielding the title compound (470 mg, 40%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + =596.

[0655] Step 2: 7-Methyl-8-[4-(pyrrolidine-1-carbonyl)phenyl]-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0656] A solution of 3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-methyl-8-[4-(pyrrolidine-1-carbonyl)phenyl]pyrazolo[1,5-a][1,3,5]triazin-4-one (530 mg, 0.890 mmol) in TFA (4 mL) was stirred for 4 h at 72 °C. The resulting mixture was concentrated under vacuum to give the title compound (560 mg, overweight). The resulting mixture was ready for use in the next step without further purification. LCMS (ES, m / z): [M+H] + =356.

[0657] Step 3: 7-Methyl-2-(methylthioalkyl)-8-[4-(pyrrolidine-1-carbonyl)phenyl]-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0658] At room temperature, methyl iodine (66 mg, 0.46 mmol) was added to a stirred mixture of 7-methyl-8-[4-(pyrrolidine-1-carbonyl)phenyl]-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (0.890 mmol) and K₂CO₃ (350 mg, 2.53 mmol) in DMF (3 mL). The resulting mixture was stirred overnight at room temperature. The mixture was filtered, and the filtrate was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 40% to 50% gradient over 5 min; detector, UV 254 nm, to give the title compound (113 mg, 34%) as a white solid. LCMS (ES, m / z): [M+H] + =370.

[0659] Step 4: 2-Methanesulfonyl-7-methyl-8-[4-(pyrrolidine-1-carbonyl)phenyl]-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0660] At 0 °C, mCPBA (105 mg, 0.606 mmol) was added to a stirred solution of 7-methyl-2-(methylthioalkyl)-8-[4-(pyrrolidine-1-carbonyl)phenyl]-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (112 mg, 0.303 mmol) in DCM (3 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution, ACN aqueous solution, 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (105 mg, 86%) as a white solid. LCMS (ES, m / z): [M+H] + =402.

[0661] Step 5: -(2,2-difluoroethoxy)-7-methyl-8-[4-(pyrrolidine-1-carbonyl)phenyl]-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0662] At 0 °C, NaH (31 mg, 0.79 mmol, 60% mineral oil) was added to a stirred solution of 2-methanesulfonyl-7-methyl-8-[4-(pyrrolidine-1-carbonyl)phenyl]-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (105 mg, 0.262 mmol) in DMF (2 mL). At 0 °C, 2,2-difluoroethanol (64 mg, 0.79 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for another 3 h. Water (10 mL) was added to the mixture. The mixture was acidified to pH 5 with 2N HCl (0.5 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with water (2 x 10 mL) and brine (10 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 10% to 50% gradient over 10 min; detector: UV 254 nm. The crude product was purified by Prep-HPLC under the following conditions (2#SHIMADZU(HPLC-01)): column: Sunfireprep C18 column, 30*150 mm, 5 µm; mobile phase: water (0.1% FA) and ACN (25% ACN to 45% over 7 min), yielding the title compound (66 mg, 63%) as a white solid. LCMS (ES, m / z): [M+H] + =404. 1 H NMR (400 MHz, DMSO-d6): δ 13.00 (s, 1H), 7.71 (d, J = 8.1 Hz, 2H), 7.59 (d, J = 8.1 Hz, 2H), 6.43 (tt, J = 53.9, 3.2 Hz, 1H), 4.70 (td, J = 15.1, 3.2 Hz, 2H), 3.48(m, 4H), 2.48 (s, 3H), 1.90 – 1.81 (m, 4H).

[0663] The examples in table Q were synthesized using synthesis method Q:

[0664] Table Q

[0665]

[0666]

[0667]

[0668]

[0669]

[0670]

[0671] Example 273-2-ethoxy-7-isopropyl-8-[3-fluoro-4-cyano-5-ethoxyphenyl]-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0672]

[0673] Treatment of 2-methanesulfonyl-7-isopropyl-8-(4-cyano-3,5-difluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (synthesized via synthetic method Q) with sodium ethoxide (5 equivalents) in DMF yielded the title compound as a white solid. LCMS (ES, m / z): [M+H] + = 386. 1H-NMR (400 MHz, DMSO-d6) δ 12.83 (s, 1H), 7.33 – 7.22 (m, 2H), 4.43 (q, J = 7.1 Hz, 2H), 4.28 (q, J = 7.0 Hz, 2H), 3.41 (septet, J = 6.8 Hz, 1H), 1.41 (t, J = 7.0 Hz, 3H), 1.36 (t, J = 7.1 Hz, 3H), 1.27 (d, J = 6.8 Hz, 6H).

[0674] Synthesis method W

[0675] Example 272-8-(4-chlorophenyl)-4-oxo-2-(prop-2-yn-1-yloxy)-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxynitrile

[0676]

[0677] Step 1: Ethyl 5-amino-4-bromo-1H-pyrazole-3-carboxylate

[0678] NBS (18.9 g, 106 mmol) was added to a stirred solution of ethyl 5-amino-1H-pyrazole-3-carboxylate (15 g, 97 mmol) in 100 mL of ACN at room temperature. The resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated under reduced pressure. This yielded the title compound (34.5 g, overweight), a dark green solid. LCMS (ES, m / z): [M+H] +=236.

[0679] Step 2: (5-amino-4-bromo-1H-pyrazole-3-yl)methanol

[0680] LiBH4 (75 mL, 150 mmol, 2 M solution in THF) was added dropwise to a stirred solution of (5-amino-4-bromo-1H-pyrazol-3-yl)methanol (97 mmol) in THF (300 mL) at 0 °C. The resulting mixture was stirred overnight at 60 °C under N2. The reaction was quenched by adding saturated NH4Cl (80 mL) at 0 °C. The resulting mixture was extracted with DCM (3 × 100 mL), and the combined organic layers were washed with brine (2 × 50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (7:1), to give the title compound (17.3 g, 93% (2 steps)) as a dark green oil. LCMS (ES, m / z): no mass ions.

[0681] Step 3: N-{[4-bromo-5-(hydroxymethyl)-2H-pyrazol-3-yl]carbamate}ethyl carbamate

[0682] At room temperature, ethyl N-thiocarbonylcarbamate (11.9 g, 90.7 mmol) was added to a stirred solution of (5-amino-4-bromo-1H-pyrazole-3-yl)methanol (17.3 g, 90.1 mmol) in DMF (90 mL). The resulting mixture was stirred at room temperature for 1 h. The mixture was then poured into ice / cold water (180 mL). The precipitated solid was collected by filtration and washed with water (3 x 20 mL). The solid was dried under vacuum. This yielded the title compound (15.8 g, 54%) as a yellow solid. LCMS (ES, m / z): [M+H] + =323, 325.

[0683] Step 4: 8-Bromo-7-(hydroxymethyl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0684] Sodium ethoxide (33.4 g, 98.3 mmol, 20 wt% in EtOH) was added to a stirred solution of N-{[4-bromo-5-(hydroxymethyl)-2H-pyrazol-3-yl]aminothioyl}carbamate (15.8 g, 48.9 mmol) in EtOH (158 mL). The reaction mixture was stirred at 80 °C for 2 h. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (100 mL). The mixture was acidified to pH 6 with 2 M HCl (40 mL) at 0 °C. The precipitated solid was collected by filtration and washed with water (3 x 20 mL). The solid was dried under vacuum. This yielded the title compound (11.5 g, 85%) as a brownish-yellow solid. LCMS (ES, m / z): [MH] - =277.

[0685] Step 5: 8-Bromo-7-(hydroxymethyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazine-4-one

[0686] At room temperature, K₂CO₃ (7.5 g, 54 mmol) and 4-methoxybenzyl chloride (8.5 g, 54 mmol) were added to a stirred solution of 8-bromo-7-(hydroxymethyl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (5.0 g, 18 mmol) in DMF (50 mL). The resulting mixture was stirred overnight at 50 °C. The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with water (3 x 30 mL) and brine (2 x 30 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (4:1), to give the title compound (3.7 g, 40%) as a brownish-yellow solid. LCMS (ES, m / z): [M+H] + =517.

[0687] Step 6: 8-(4-chlorophenyl)-7-(hydroxymethyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazine-4-one

[0688] At room temperature, water (3 mL), K₂CO₃ (1.71 g, 12.4 mmol), and Pd(Dtbpf)Cl₂ (414 mg, 0.635 mmol) were added to a stirred mixture of 8-bromo-7-(hydroxymethyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazine-4-one (3.2 g, 6.2 mmol) and p-chlorophenylboronic acid (1.93 g, 12.4 mmol) in dioxane (30 mL). The resulting mixture was stirred overnight at 80 °C under N₂. The resulting mixture was filtered, and the filter cake was washed with 1,4-dioxane (3 x 5 mL). The resulting mixture was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 50% to 70% gradient over 10 min; detector: UV 254 nm. This yielded the title compound (1.3 g, 38%) as a brownish-yellow oil. LCMS (ES, m / z): [M+H] + =549.

[0689] Step 7: 8-(4-chlorophenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-4-oxopyrazolo[1,5-a][1,3,5]triazine-7-carboxaldehyde

[0690] At 0 °C, 4.2 g (9.9 mmol) of Des Martin periodane was added to a stirred solution of 8-(4-chlorophenyl)-7-(hydroxymethyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazine-4-one (1.3 g, 2.4 mmol) in THF (13 mL). The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched at 0 °C by adding saturated sodium bicarbonate (30 mL). The resulting mixture was extracted with EA (3 x 30 mL), and the combined organic layers were washed with brine (2 x 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (4:1), to give the title compound (1.08 g, 83%) as a yellow solid. LCMS (ES, m / z): [M+H] + =547.

[0691] Step 8: 8-(4-chlorophenyl)-7-[(1E)-(hydroxyimino)methyl]-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazine-4-one

[0692] At room temperature, 8-(4-chlorophenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-4-oxopyrazolo[1,5-a][1,3,5]triazine-7-carboxaldehyde (1.08 g, 1.97 mmol) and NH₂OH·HCl (274 mg, 3.95 mmol) were added to a mixture of AcOH (10.8 mL) and water (3.6 mL). The resulting mixture was stirred at 60 °C for 4 h. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 30% to 60% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (882 mg, 79%) as a yellow solid. LCMS (ES, m / z): [M+H] + =562.

[0693] Step 9: 8-(4-chlorophenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-4-oxopyrazolo[1,5-a][1,3,5]triazine-7-carboxynitrile

[0694] At room temperature, TEA (1.6 g, 16 mmol) and TFAA (3.3 g, 16 mmol) were added to a stirred solution of 8-(4-chlorophenyl)-7-[(1E)-(hydroxyimino)methyl]-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}pyrazolo[1,5-a][1,3,5]triazine-4-one (882 mg, 1.57 mmol) in DCM (89 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was diluted with water (30 mL). The resulting mixture was extracted with DCM (3 × 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (1.5 g, overweight) as a yellow solid. LCMS (ES, m / z): [M+H] + =544.

[0695] Step 10: 8-(4-chlorophenyl)-4-oxo-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxynitrile

[0696] A solution of 8-(4-chlorophenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-4-oxopyrazolo[1,5-a][1,3,5]triazine-7-carboxynitrile (1.5 g, 2.8 mmol) in TFA (15 mL) was stirred at 80 °C for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 50% to 70% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (547 mg, 65%) as a brownish-yellow oil. LCMS (ES, m / z): [MH] - =302.

[0697] Step 11: 8-(4-chlorophenyl)-2-(methylthioalkyl)-4-oxo-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxynitrile

[0698] At room temperature, K₂CO₃ (670 mg, 4.85 mmol) and MeI (229 mg, 1.62 mmol) were added to a stirred solution of 8-(4-chlorophenyl)-4-oxo-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxynitrile (491 mg, 1.62 mmol) in DM (10 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with water (2 x 20 mL) and brine (2 x 20 mL) and dried over anhydrous Na₂SO₄. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 40% to 80% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (168 mg, 33%) as a yellow solid. LCMS (ES, m / z): [MH] - =316.

[0699] Step 12: 8-(4-chlorophenyl)-2-methanesulfonyl-4-oxo-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxylonitrile and 8-(4-chlorophenyl)-2-(methylsulfinyl)-4-oxo-3,4-dihydropyrazolo[1,5-a][1,3,5]triazine-7-carboxylonitrile

[0700] mCPBA (183 mg, 1.061 mmol) was added to a stirred solution of 8-(4-chlorophenyl)-2-(methylthioalkyl)-4-oxo-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxynitrile (168 mg, 0.529 mmol) in DCM (4 mL) at 0 °C. The resulting mixture was stirred at 0 °C for 15 min and then at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 50% to 80% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (170 mg, 92%) as a yellow solid. LCMS (ES, m / z): [MH] - =332 and 348.

[0701] Step 13: 8-(4-chlorophenyl)-4-oxo-2-(prop-2-yn-1-yloxy)-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxynitrile

[0702] At 0 °C, NaH (25 mg, 0.62 mmol, 60% mineral oil) was added to a stirred solution of 2,2-difluoroethanol (33.8 mg, 0.412 mmol) in DMF (4 mL). The resulting mixture was stirred at 0 °C for 30 min. At room temperature, 8-(4-chlorophenyl)-2-methanesulfonyl-4-oxo-3H-pyrazolo[1,5-a][1,3,5]triazine-7-carboxylonitrile and 8-(4-chlorophenyl)-2-(methylsulfinyl)-4-oxo-3,4-dihydropyrazolo[1,5-a][1,3,5]triazine-7-carboxylonitrile (72 mg, 0.206 mmol, Mw: 349.0) were added to the mixture. The resulting mixture was stirred at room temperature for another 2 h. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with water (2 x 10 mL) and brine (10 mL) and dried over anhydrous Na₂SO₄. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 30% to 80% gradient over 10 min; detector, UV 254 nm, to obtain the crude product. The crude product was purified by Prep-HPLC under the following conditions (column: Xselect CSH C18 OBD column 30*150 mm 5 μm, n; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 46% B to 57% B, 57% B over 10 min; wavelength: 254 / 220 nm; RT1 (min): 8.30), to give the title compound (23.8 mg, 27%) as a white solid. LCMS (ES, m / z): [MH] - =350. 1 H NMR-371-0: (400 MHz, DMSO-d6) δ 7.90 – 7.88 (m, 2H), 7.62 –7.60 (m, 2H), 6.58 – 6.31 (tt, J = 53.7, 3.1 Hz, 1H), 4.78 – 4.70 (td, J =15.2, 3.1 Hz, 2H).

[0703] The examples in Table W were synthesized using synthesis method W:

[0704] Table W

[0705]

[0706] Synthesis method P

[0707] Examples 276 and 277 - 2-(2,2-difluoroethoxy)-7-[oxacyclohexane-2-yl]-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0708]

[0709] Step 1: 3-(oxacyclohexane-2-yl)-3-oxo-2-(3,4,5-trifluorophenyl)propionitrile

[0710] At 0 °C, NaH (1.40 g, 35.1 mmol, 60% in mineral oil) was added fractionally to a stirred solution of 2-(3,4,5-trifluorophenyl)acetonitrile (3.0 g, 18 mmol) in THF (30 mL). The resulting mixture was stirred at 0 °C for 30 min. At 0 °C, methyl oxane-2-carboxylate (7.58 g, 52.6 mmol) was added to the mixture. The resulting mixture was stirred overnight at room temperature. The reaction was quenched by adding 60 mL of water at 0 °C. The resulting mixture was extracted with EA (3 x 60 mL). The combined organic layers were washed with brine (2 x 60 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1), to give the title compound (4.5 g, 88%) as a brown oil. LCMS (ES, m / z): [MH] - =282.

[0711] Step 2: 5-(oxacyclohexane-2-yl)-4-(3,4,5-trifluorophenyl)-2H-pyrazole-3-amine

[0712] At room temperature, hydrazine hydrate (1.99 g, 31.8 mmol, 80 wt%) and TFA (3.6 g, 32 mmol) were added to a stirred solution of 3-(oxacyclohexan-2-yl)-3-oxo-2-(3,4,5-trifluorophenyl)propionitrile (4.5 g, 16 mmol) in EtOH (45 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure. The mixture was diluted with water (80 mL). The mixture was extracted with EA (3 x 80 mL). The combined organic layers were washed with brine (2 x 80 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1), to give the title compound (4.6 g, 97%) as a brown oil. The crude product mixture could be used directly for the next step without further purification. LCMS (ES, m / z): [M+H]+ =298.

[0713] Step 3: N-{[5-(oxacyclohexane-2-yl)-4-(3,4,5-trifluorophenyl)-2H-pyrazol-3-yl]carbamate}ethyl carbamate

[0714] At room temperature, ethyl N-thiocarbonylcarbamate (2.03 g, 15.5 mmol) was added to a stirred solution of 5-(oxacyclohexane-2-yl)-4-(3,4,5-trifluorophenyl)-2H-pyrazole-3-amine (4.6 g, 15 mmol) in DMF (46 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was poured into water (80 mL) at 0 C. The precipitated solid was collected by filtration and washed with water (2 x 10 mL). The solid was dried under vacuum. This yielded the title compound (5.5 g, 83%) as a yellow solid. The crude product was used directly for the next step without further purification. LCMS (ES, m / z): [M+H] + =429.

[0715] Step 4: 7-(oxacyclohexane-2-yl)-2-thioalkyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0716] A solution of N-{[5-(oxacyclohexane-2-yl)-4-(3,4,5-trifluorophenyl)-2H-pyrazol-3-yl]carbamate (5.5 g, 13 mmol) in 2M NaOH (55 mL) was stirred at room temperature for 1 h. The mixture was acidified to pH 6 with 2M HCl (25 mL). The precipitated solid was collected by filtration and washed with water (2 x 10 mL). The solid was dried under vacuum. This yielded the title compound (4.7 g, 96%) as a grayish-white solid. The crude product was used directly for the next step without further purification. LCMS (ES, m / z): [M+H] + =383.

[0717] Step 5: 2-(methylthioalkyl)-7-(oxacyclohexane-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0718] At room temperature, K₂CO₃ (5.10 g, 36.9 mmol) and MeI (1.40 g, 9.83 mmol) were added to a stirred solution of 7-(oxan-2-yl)-2-thioalkyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (4.7 g, 12 mmol) in DMF (47 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was poured into ice / water (80 mL). The precipitated solid was collected by filtration and washed with water (2 x 10 mL). The resulting solid was dried under vacuum. This yielded the title compound (5.4 g, overweight) as a grayish-white solid. The crude product was used directly for the next step without further purification. LCMS (ES, m / z): [M+H] + =397.

[0719] Step 6: 2-Methanesulfonyl-7-(oxacyclohexane-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0720] At 0 °C, mCPBA (4.70 g, 27.2 mmol) was added in portions to a stirred solution of 2-(methylthioalkyl)-7-(oxecyclohexane-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (12 mmol) in DCM (54 mL). The resulting mixture was stirred at 0 °C for 15 min and then at room temperature for 2 h, followed by concentration under reduced pressure. The resulting mixture was diluted with water (80 mL). The resulting mixture was extracted with EA (3 x 80 mL). The combined organic layers were washed with brine (2 x 80 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (9 g, overweight) as a yellow solid. The crude product was used directly for the next step without further purification. LCMS (ES, m / z): [M+H] + =429.

[0721] Step 7: 2-(2,2-difluoroethoxy)-7-[oxacyclohexane-2-yl]-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (enantiomer 1: Example 276, enantiomer 2: Example 277)

[0722] At 0 °C, NaH (77 mg, 1.9 mmol, 60% mineral oil) was added to a stirred solution of 2,2-difluoroethanol (236 mg, 2.88 mmol) in DMF (8 mL). The resulting mixture was stirred at 0 °C for 30 min. 2-Methanesulfonyl-7-(1-methoxypropyl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (400 mg, 0.961 mmol) was added to the mixture at room temperature. The resulting mixture was stirred at room temperature for another 2 h. The mixture was acidified to pH 7 with 2M HCl (0.5 mL). The resulting mixture was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 50% to 80% gradient over 10 min; detector, UV 254 nm. The racemic mixture (180 mg) was purified by SFC under the following conditions (column: (S, S)-Whelk-O 1 5 μm Kromasil, 3*25 cm, 5 μm; mobile phase A: CO2, mobile phase B: MeOH-HPLC; flow rate: 100 mL / min; gradient: isocratic 55% B; column temperature (°C): 35; back pressure (bar): 100; wavelength: 210 / 276 nm; RT1 (min): 2.8; RT2 (min): 4.2; sample solvent: MeOH-HPLC; injection volume: 1 mL), yielding the title compound (enantiomer 1: 70 mg) with RT1 (min): 2.8 and the title compound (enantiomer 2, 90 mg) with RT2 (min): 4.2, as white solids. The crude enantiomer 1 (70 mg) was purified by Prep-HPLC under the following conditions (column: Sunfire prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 58% B to 73% B over 8 min; wavelength: 254 / 220 nm; RT1 (min): 5.50) to give the title compound (enantiomer 1, 28 mg, 7%) as a white solid. LCMS (ES, m / z): [M+H] + =431. 1H NMR (400 MHz, DMSO-d6) δ 13.23 (s, 1H), 7.68 – 7.62 (m, 2H), 6.45 (tt, J = 53.9, 3.0 Hz, 1H), 4.71 (tdd, J = 15.2, 4.8, 2.9 Hz, 2H), 4.54 (dd, J = 11.0, 2.3 Hz, 1H), 4.01(d, J = 11.2 Hz, 1H), 3.65 – 3.54 (m, 1H), 2.09 – 2.06 (qm, 1H), 1.92 – 1.90(m, 1H), 1.78 – 1.75 (m, 1H), 1.64 – 1.55 m, 3H). The crude enantiomer 2 product (90 mg) was purified by Prep-HPLC under the following conditions (column: Xselect CSH C18 OBD column 30*150 mm 5 μm, n; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 49% B to 65% B over 8 min; wavelength: 254 / 220 nm; RT1 (min): 7.25), yielding the title compound (49 mg, 12%) as a white solid. LCMS (ES, m / z): [M+H] + =431. 1 H NMR (400 MHz, DMSO-d6) δ 13.25 (s, 1H), 7.68 – 7.63 (m, 2H), 6.44 (tt,J = 53.8, 3.1 Hz, 1H), 4.72 (tdd, J = 15.2, 4.9, 2.9 Hz, 2H), 4.53 (dd, J =11.1, 2.3 Hz, 1H), 4.01 (d, J = 11.2 Hz, 1H), 3.62 – 3.56 (m, 1H), 2.09 –2.01 (m, 1H), 2.00– 1.90 (m, 1H), 1.78 – 1.75 (m, 1H), 1.64 – 1.58 (m, 3H).

[0723] The examples in Table P were synthesized using synthesis method P:

[0724] Table P

[0725]

[0726]

[0727]

[0728] Example 288-8-(4-chlorophenyl)-2-(difluoromethoxy)-7-(oxacyclohexane-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0729]

[0730] 8-(4-chlorophenyl)-2-methanesulfonyl-7-(oxacyclohexane-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one was synthesized using synthetic method E.

[0731] Step 1: 8-(4-chlorophenyl)-2-hydroxy-7-(oxacyclohexane-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0732] At room temperature, 2 M NaOH (5 mL) was added to a stirred solution of 8-(4-chlorophenyl)-2-methanesulfonyl-7-(oxacyclohexane-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (synthesized using synthetic method P; 1.0 g, 2.4 mmol) in dioxane (10 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was acidified to pH 5 with 2 M HCl (6 mL). The resulting mixture was extracted with EA (3 x 30 mL). The combined organic layers were washed with brine (30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 50% to 90% gradient over 10 min; detector, UV 254 nm. This yielded the title compound (0.52 g, 62%) as a yellow solid. LCMS (ES, m / z): [M+H] + =347.

[0733] Step 2: 8-(4-chlorophenyl)-2-(difluoromethoxy)-7-(oxacyclohexane-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0734] At room temperature, ethyl 2-bromo-2,2-difluoroacetate (293 mg, 1.44 mmol) and Cs₂CO₃ (705 mg, 2.16 mmol) were added to a stirred solution of 8-(4-chlorophenyl)-2-hydroxy-7-(oxacyclohexane-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (250 mg, 0.721 mmol) in DMF (2 mL). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with water (10 mL). The mixture was acidified to pH 5 with 2 M HCl. The resulting mixture was extracted with EA (3 x 15 mL). The combined organic layers were washed with water (2 x 15 mL) and brine (15 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 50% to 100% gradient over 10 min; detector: UV 254 nm. The crude product was purified by Prep-HPLC under the following conditions (column: Xselect CSH C18 OBD column 30*150 mm 5 μm, n; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 57% B to 74% B, 74% B over 9 min; wavelength: 254 / 220 nm; RT1 (min): 5.23), yielding the title compound (8.7 mg, 3%) as a white solid. LCMS (ES, m / z): [M+H] + = 397. 1H NMR (400MHz, DMSO-d6) δ 7.72 (t, J = 76.0 Hz, 1H), 7.72 – 7.69 (m, 2H), 7.55 – 7.49(m, 2H), 4.50 (dd, J = 11.1, 2.2 Hz, 1H), 3.99 – 3.96 (m, 1H), 3.59 – 3.53 (m, 1H), 2.10 – 2.00 (m, 1H), 1.92 – 1.87 (m, 1H), 1.77 – 1.69 (m, 1H), 1.63 – 1.54 (m, 3H).

[0735] Example 289 was synthesized using the same method:

[0736]

[0737] LCMS (ES, m / z): [M+H] += 390. 1H NMR (400 MHz, DMSO-d6) δ 8.60 (m,1H), 8.05 – 8.01 (m, 1H), 7.91 – 7.74 (m, 2H), 7.56 – 7.52 (m, 1H), 7.43 –7.14 (m, 4H).

[0738] Example 290 was synthesized using the same method:

[0739]

[0740] LCMS (ES, m / z): [M+H] + = 375. 1H NMR (400 MHz, DMSO-d6) δ 7.72 (t, J = 70.3 Hz, 1H), 7.48 – 7.42 (m, 2H), 3.34 (septet, J = 6.8 Hz, 1H), 1.31 (d, J = 6.7 Hz, 6H).

[0741] Synthesis method Z

[0742] Example 291-7-(4-fluorophenyl)-6-methyl-2-(prop-2-yn-1-ylthioalkyl)-3H-imidazo[2,1-f][1,2,4]triazin-4-one

[0743]

[0744] Step 1: Ethyl 4-methyl-1H-imidazol-2-carboxylate

[0745] At room temperature, ethyl glyoxylate (14.2 g, 139 mmol, 50% wt / w toluene) and NH3-H2O (19.5 g, 555 mmol) were added to a stirred solution of methylglyoxal (10.0 g, 139 mmol, 40% wt / w in water) in MeOH (100 mL). The resulting mixture was stirred overnight at room temperature. The mixture was concentrated under reduced pressure. The mixture was diluted with water (200 mL). The mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with brine (2 x 50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% NH3-H2O). 3.H₂O), gradient 10% to 50% over 10 min; detector, UV 254 nm, yielded the title compound (3.0 mg, 14%) as a purple solid. LCMS (ES, m / z): [M+H] + =155.

[0746] Step 2: Ethyl 1-amino-4-methylimidazolium-2-carboxylate

[0747] At room temperature and under N2, t-BuOK (1.57 g, 14.0 mmol) was added to a stirred solution of ethyl 4-methyl-1H-imidazolium-2-carboxylate (1.80 g, 11.7 mmol) in DMF (20 mL). The resulting mixture was stirred at room temperature for 30 min. At room temperature, amino-4-nitrobenzene ester (4.25 g, 23.4 mmol) was added to the mixture. The resulting mixture was stirred overnight at room temperature. The resulting mixture was diluted with water (50 mL). The mixture was alkalized to pH 9 with saturated NaHCO3 (aqueous solution). The resulting mixture was extracted with DCM (3 x 20 mL). The combined organic layers were washed with brine (2 x 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% NH3·H2O) in a 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (900 mg, 46%) as a yellow solid. LCMS (ES, m / z): [M+H] + =170.

[0748] Step 3: Ethyl 1-amino-5-bromo-4-methylimidazolium-2-carboxylate

[0749] NBS (463 mg, 2.60 mmol) was added to a stirred solution of ethyl 1-amino-4-methylimidazolium-2-carboxylate (400 mg, 2.36 mmol) in ACN (8 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% NH3·H2O) in a 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (120 mg, 21%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =248.

[0750] Step 4: Ethyl 1-amino-5-(4-fluorophenyl)-4-methylimidazolium-2-carboxylate

[0751] At room temperature under N2, Pd(PPh3)4 (93 mg, 0.081 mmol) and K2CO3 (223 mg, 1.61 mmol) were added to a stirred mixture of ethyl 1-amino-5-bromo-4-methylimidazolium-2-carboxylate (200 mg, 0.806 mmol) and 4-fluorophenylboronic acid (226 mg, 1.61 mmol) in dioxane (4 mL) and water (0.4 mL). The final reaction mixture was irradiated with microwave at 110 °C for 2 h. The mixture was then cooled to room temperature. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with EA (3 x 10 mL). The combined organic layers were washed with brine (2 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 10% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (120 mg, 56%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =264.

[0752] Step 5: 1-({[(ethoxycarbonyl)amino]methionyl}amino)-5-(4-fluorophenyl)-4-methylimidazolium-2-carboxylic acid ethyl ester

[0753] At room temperature, ethyl N-thiocarbonylcarbamate (66 mg, 0.50 mmol) was added to a stirred solution of ethyl 1-amino-5-(4-fluorophenyl)-4-methylimidazolium-2-carboxylate (120 mg, 0.456 mmol) in DMF (2 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 5 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure to give the title compound (130 mg, 72%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =395.

[0754] Step 6: 7-(4-fluorophenyl)-6-methyl-2-thioalkyl-3H-imidazo[2,1-f][1,2,4]triazine-4-one

[0755] At room temperature, ethyl 1-({[(ethoxycarbonyl)amino]methionyl}amino)-5-(4-fluorophenyl)-4-methylimidazolium-2-carboxylate (130 mg, 0.330 mmol) in water (2 mL) was added to a stirred solution of 2 mL. The resulting mixture was stirred at 85 °C for 1 h. The mixture was then cooled to room temperature. The mixture was acidified to pH 5 with concentrated HCl. The precipitated solid was collected by filtration and washed with water (3 x 1 mL). The resulting solid was dried under vacuum to give the title compound (50 mg, 55%) as a white solid. LCMS (ES, m / z): [M+H]+ = 277.

[0756] Step 7: -(4-fluorophenyl)-6-methyl-2-(prop-2-yn-1-ylthioalkyl)-3H-imidazo[2,1-f][1,2,4]triazin-4-one

[0757] At room temperature and under N2, propargyl bromide (8.6 mg, 0.072 mmol) was added to a stirred mixture of 7-(4-fluorophenyl)-6-methyl-2-thioalkyl-3H-imidazo[2,1-f][1,2,4]triazin-4-one (50 mg, 0.18 mmol) and K2CO3 (50 mg, 0.36 mmol) in DMF (1 mL). The resulting mixture was stirred at room temperature and under N2 for 1 h. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EA (3 x 5 mL). The combined organic layers were washed with brine (2 x 5 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: Sunfire Prep C18 column, 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 32% B to 58% B, 58% B over 7 min; wavelength: 254 / 220 nm; RT1 (min): 5.60), yielding the title compound (7 mg, 12%) as a white solid. LCMS (ES, m / z): [M+H] + =315. 1H NMR (400 MHz, DMSO-d6) δ 12.80 (s, 1H), 7.84 – 7.75 (m, 2H), 7.40 – 7.31 (m, 2H), 3.91 (d, J =2.6 Hz, 2H), 3.28 (t, , J = 2.6 Hz, 1H), 2.39 (s, 3H).

[0758] Examples from Table Z were synthesized using synthesis method Z:

[0759] Table Z

[0760]

[0761] Example 296-7-(4-chlorophenyl)-2-(2,2-difluoroethoxy)-6-isopropyl-3H-imidazo[2,1-f][1,2,4]triazine-4-one

[0762]

[0763] Step 1: 3-ethyl 1-aminoimidazole-2-carboxylic acid

[0764] At 0 °C, NaH (5.71 g, 143 mmol, 60% in mineral oil) was added fractionally to a stirred solution of ethyl 1H-imidazolium-2-carboxylate (10 g, 71 mmol) in THF (200 mL). The resulting mixture was stirred at room temperature for 2 h. At room temperature, aminodiphenylphosphonite (33.3 g, 143 mmol) was added to the mixture. The resulting mixture was stirred again overnight at 65 °C. The mixture was cooled to room temperature. The reaction was quenched by adding 30 mL of water at 0 °C. The resulting mixture was filtered, and the filter cake was washed with THF (3 x 30 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1), to give the title compound (6.0 g, 54%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =156.

[0765] Step 2: ethyl 1-[(tert-Butoxycarbonyl)amino]imidazolium-2-carboxylate

[0766] At room temperature, di-tert-butyl dicarbonate (10.1 g, 46.4 mmol) and DMAP (0.94 g, 7.7 mmol) were added to a stirred solution of ethyl 1-aminoimidazolium-2-carboxylate (6.0 g, 39 mmol) in THF (120 mL). The resulting mixture was stirred overnight at 65 °C. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1), to give the title compound (3.4 g, 34.%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =256.

[0767] Step 3: Ethyl 4-bromo-1-[(tert-Butoxycarbonyl)amino]imidazolium-2-carboxylate

[0768] At 0 °C, NBS (1.26 g, 7.05 mmol) was added fractionally to a stirred solution of ethyl 1-[(tert-butyloxycarbonyl)amino]imidazolium-2-carboxylate (3.0 g, 12 mmol) in ACN (60 mL). The resulting mixture was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1), to give the title compound (2.0 g, 51%) as a yellow oil. LCMS (ES, m / z): [M+H] + =334.

[0769] Step 4: 1-[(tert-Butoxycarbonyl)amino]-4-(prop-1-en-2-yl)imidazol-2-carboxylic acid ethyl ester

[0770] At room temperature and under N2, Pd(PPh3)4 (0.69 g, 0.60 mmol) and K2CO3 (1.65 g, 12.0 mmol) were added to a stirred solution of ethyl 4-bromo-1-[(tert-butoxycarbonyl)amino]imidazolium-2-carboxylate (2.0 g, 6.0 mmol) and 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxane (4.02 g, 23.9 mmol) in dioxane (40 mL) and water (4 mL). The resulting mixture was stirred overnight at 90 °C under N2. The mixture was then cooled to room temperature. The resulting mixture was diluted with water (80 mL). The resulting mixture was extracted with EA (3 x 20 mL). The combined organic layers were washed with brine (2 x 20 mL) and subjected to anhydrous Na2SO4. 4干 Dry. After filtration, concentrate the filtrate under reduced pressure. Purify the residue by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), in a 40% to 60% gradient over 10 min; detector, UV 254 nm, to give the title compound (600 mg, 34%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =296.

[0771] Step 5: 1-[(tert-Butoxycarbonyl)amino]-4-isopropylimidazol-2-carboxylic acid ethyl ester

[0772] At room temperature under N2, 10% Pd / C (300 mg, 2.82 mmol) was added to a stirred solution of ethyl 1-[(tert-butoxycarbonyl)amino]-4-(prop-1-en-2-yl)imidazolium-2-carboxylate (600 mg, 2.03 mmol) in MeOH (12 mL). The resulting mixture was stirred overnight at room temperature under nitrogen atmosphere. The mixture was filtered, and the filter cake was washed with MeOH (5 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (5:1), to give the title compound (500 mg, 83%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =298.

[0773] Step 6: Ethyl 1-amino-4-isopropylimidazolium-2-carboxylate

[0774] At room temperature, 3.3 mL of TFA was added to a stirred solution of ethyl 1-[(tert-butyloxycarbonyl)amino]-4-isopropylimidazolium-2-carboxylate (500 mg, 1.68 mmol) in 10 mL of DCM. The resulting mixture was stirred at room temperature for 1 h. The mixture was then concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA) in a 10% to 30% gradient over 10 min; detector, UV 254 nm, to give the title compound (300 mg, 91%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + =198.

[0775] Step 7: Ethyl 1-amino-5-bromo-4-isopropylimidazolium-2-carboxylate

[0776] NBS (248 mg, 1.40 mmol) was added to a stirred solution of ethyl 1-amino-4-isopropylimidazolium-2-carboxylate (250 mg, 1.27 mmol) in ACN (6 mL) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The mixture was then concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% NH3). . H₂O), gradient 30% to 50% over 10 min; detector, UV 254 nm, yielded the title compound (200 mg, 57%) as a yellow solid. LCMS (ES, m / z): [M+H] + =276.

[0777] Step 8: Ethyl 1-amino-5-(4-chlorophenyl)-4-isopropylimidazolium-2-carboxylate

[0778] At room temperature under N2, Pd(PPh3)4 (63 mg, 0.054 mmol) and K2CO3 (150 mg, 1.09 mmol) were added to a solution of ethyl 1-amino-5-bromo-4-isopropylimidazolium-2-carboxylate (150 mg, 0.543 mmol) and p-chlorophenylboronic acid (170 mg, 1.09 mmol) in dioxane (3 mL) and water (0.3 mL). The resulting mixture was stirred at 90 °C under N2 for 3 h. The mixture was then cooled to room temperature. The resulting mixture was filtered, and the filter cake was washed with 1,4-dioxane (3 x 1 mL). The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), in a 40% to 60% gradient over 10 min; detector, UV 254 nm, to give the title compound (110 mg, 66%) as a brown solid. LCMS (ES, m / z): [M+H] + =308.

[0779] Step 9: 5-(4-chlorophenyl)-1-({[(ethoxycarbonyl)amino]methionyl}amino)-4-isopropylimidazol-2-carboxylic acid ethyl ester

[0780] At room temperature, N-thiocarbonylcarbamate (52 mg, 0.39 mmol) was added to a stirred solution of ethyl 1-amino-5-(4-chlorophenyl)-4-isopropylimidazolium-2-carboxylate (110 mg, 0.357 mmol) in DMF (2 mL). The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was dissolved in water (5 mL) at 0 °C. The precipitated solid was collected by filtration and washed with water (2 x 1 mL). The resulting solid was dried under vacuum to give the title compound (90 mg, 57%) as a yellow solid. LCMS (ES, m / z): [M+H] + =439.

[0781] Step 10: 7-(4-chlorophenyl)-6-isopropyl-2-thioalkyl-3H-imidazo[2,1-f][1,2,4]triazine-4-one

[0782] A solution of ethyl 5-(4-chlorophenyl)-1-({[(ethoxycarbonyl)amino]methionyl}amino)-4-isopropylimidazolium-2-carboxylate (90 mg, 0.21 mmol) in 2M NaOH (2 mL) at room temperature. The resulting mixture was stirred at 85 °C for 2 h. The mixture was then cooled to room temperature. The resulting mixture was diluted with water (10 mL). The mixture was acidified to pH 5 with 2M HCl (3 mL). The precipitated solid was collected by filtration and washed with water (3 x 2 mL). The resulting solid was dried under vacuum to give the title compound (60 mg, 91%) as a yellow solid. LCMS (ES, m / z): [M+H] + =321.

[0783] Step 11: 7-(4-chlorophenyl)-6-isopropyl-2-(methylthioalkyl)-3H-imidazo[2,1-f][1,2,4]triazine-4-one

[0784] MeI (24 mg, 0.17 mmol) and K₂CO₃ (78 mg, 0.56 mmol) were added to a stirred solution of 7-(4-chlorophenyl)-6-isopropyl-2-thioalkyl-3H-imidazo[2,1-f][1,2,4]triazine-4-one (90 mg, 0.28 mmol) in DMF (2 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The mixture was filtered, and the filter cake was washed with DMF (2 x 1 mL). The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 40% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (50 mg, 53%) as a white solid. LCMS (ES, m / z): [M+H] + =335.

[0785] Step 12: 7-(4-chlorophenyl)-6-isopropyl-2-methanesulfonyl-3H-imidazo[2,1-f][1,2,4]triazine-4-one

[0786] At room temperature, mCPBA (52 mg, 0.30 mmol) was added to a stirred solution of 7-(4-chlorophenyl)-6-isopropyl-2-(methylthioalkyl)-3H-imidazo[2,1-f][1,2,4]triazin-4-one (50 mg, 0.15 mmol) in DCM (1 mL). The resulting mixture was stirred overnight at room temperature. The filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography using C18 silica gel; ACN aqueous solution (0.1% FA), 40% to 50% gradient over 10 min; detector, UV 254 nm, to give the title compound (40 mg mixture, 73%) as a white solid. LCMS (ES, m / z): [M+H] + =367.

[0787] Step 13: 7-(4-chlorophenyl)-2-(2,2-difluoroethoxy)-6-isopropyl-3H-imidazo[2,1-f][1,2,4]triazine-4-one

[0788] At 0 °C, NaH (6.5 mg, 0.16 mmol, 60% mineral oil) was added to a stirred solution of 2,2-difluoroethanol (13.4 mg, 0.164 mmol) in DMF (1 mL). The resulting mixture was stirred at 0 °C for 30 min. At 0 °C, 7-(4-chlorophenyl)-6-isopropyl-2-methanesulfonyl-3H-imidazo[2,1-f][1,2,4]triazine-4-one (30 mg, 0.082 mmol) was added to the above mixture. The resulting mixture was stirred at 50 °C for another 2 h. The mixture was then cooled to room temperature. The mixture was neutralized to pH 7 with 2M HCl (0.1 mL). The mixture was purified by reversed-phase rapid chromatography under the following conditions: column, C18 silica gel; mobile phase, aqueous ACN (0.1% FA), 40% to 50% gradient over 10 min; detector, UV 254 nm. The residue was obtained and further purified by Prep-HPLC under the following conditions (column: Xselect CSH C18 OBD column 30*150mm 5μm, n; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 40% B to 52% B, 52% B over 10 min; wavelength: 254 / 220 nm; RT1 (min): 9.35), yielding the title compound (8 mg, 27%) as a white solid. LCMS (ES, m / z): [M+H] + =369. 1¹H NMR (400 MHz, DMSO-d⁶) δ 12.86 (s, 1H), 7.64–7.57 (m, 4H), 6.39 (tt, J = 3.2 Hz, 1H), 4.49 (td, J = 15.0, 3.2 Hz, 2H), 3.10 (septet, J = 6.7 Hz, 1H), 1.23 (d, J = 6.8 Hz, 6H).

[0789] Example 297 was synthesized using the same procedure:

[0790]

[0791] LCMS (ES, m / z): [M+H] + = 343. 1H NMR (400 MHz, DMSO-d6) δ 12.74 (s, 1H), 7.67 – 7.64 (m, 2H), 7.59 – 7.57 (m, 2H), 4.90 (d, J = 2.5 Hz, 2H), 3.73 (d, J = 2.6 Hz, 1H), 3.10 (septet J = 6.7 Hz, 1H), 1.23 (d, J = 6.8 Hz, 6H).

[0792] Example 341 - 8-Methyl-7-phenyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (Example 11388)

[0793]

[0794] Step 1: 2-Methyl-3-oxo-3-phenylpropionitrile

[0795] At 0 °C, NaH (1.2 g, 30 mmol, 60% in mineral oil) was added dropwise to a stirred solution of benzoylacetonitrile (4.35 g, 30.0 mmol) in DMF (43.5 mL). The resulting mixture was stirred at 0 °C for 15 min, followed by dropwise addition of CH3I (4.25 g, 30.0 mmol) at 0 °C. The mixture was stirred at room temperature for 1 h. The reaction was quenched with water (50 mL) and extracted with EA (3 x 40 mL). The combined organic layers were washed with brine (40 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (8:2) to give the title compound (2.77 g, 58%) as a yellow oil. LCMS (ES, m / z): [MH] - =158.

[0796] Step 2: 4-Methyl-5-phenyl-2H-pyrazole-3-amine

[0797] At room temperature, hydrazine hydrate (1.45 g, 28.9 mmol, 80 wt%) was added to a stirred solution of 2-methyl-3-oxo-3-phenylpropionitrile (2.3 g, 14 mmol) in EtOH (23 mL). The resulting mixture was stirred overnight at 80 °C. The mixture was concentrated under reduced pressure and the residue was diluted with water (50 mL). The mixture was extracted with EA (3 x 50 mL). The combined organic layers were washed with brine (30 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. This yielded the title compound (2.2 g, 88%) as a brown oil. The crude product was used directly for the next step without further purification. LCMS (ES, m / z): [M+H] + =174.

[0798] Step 3: N-[(4-methyl-5-phenyl-2H-pyrazol-3-yl)carbamoyl]carbamate

[0799] At room temperature, ethyl N-thiocarbonylcarbamate (1.66 g, 12.7 mmol) was added to a stirred solution of 4-methyl-5-phenyl-2H-pyrazole-3-amine (2.2 g, 13 mmol) in DMF (48 mL). The resulting mixture was stirred at room temperature for 1 h. The mixture was then poured into ice / water (350 mL). The precipitated solid was collected by filtration and washed with water (3 x 20 mL). The resulting solid was dried under reduced pressure. This yielded the precipitate of the title compound (3.3 g, 85%) as a yellow solid. LCMS (ES, m / z): [M+H] +=305.

[0800] Step 4: 8-Methyl-7-phenyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0801] A solution of Na (0.76 g, 33 mmol) in EtOH (64 mL) was stirred at 0 °C for 1 h. N-[(4-methyl-5-phenyl-2H-pyrazol-3-yl)aminothioyl]carbamate (5.0 g, 16 mmol) was added to the mixture at room temperature. The resulting mixture was stirred at 80 °C for another 1 h. The mixture was filtered, and the filter cake was dissolved in water (50 mL). The mixture was acidified to pH 6 with 1N HCl (35 mL). The precipitated solid was collected by filtration and washed with water (3 x 10 mL). The resulting solid was dried under reduced pressure. This yielded compound 8 (2.9 g, 74%) as a yellow solid. The crude product was ready for use in the next step without further purification. LCMS (ES, m / z): [M+H] + =259.

[0802] Step 5: 8-Methyl-7-phenyl-2-(prop-2-yn-1-ylthioalkyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0803] At room temperature, propargyl bromide (59 mg, 0.50 mmol) was added to a stirred mixture of 8-methyl-7-phenyl-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (320 mg, 1.24 mmol) and K2CO3 (514 mg, 3.72 mmol) in DMF (5 mL). The resulting mixture was stirred at room temperature for 1 h. The mixture was filtered, and the filtrate was purified by Prep-HPLC under the following conditions (column: SunFire Prep C18 OBD column, 19*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 38% B to 58% B over 8 min; wavelength: 220 nm; RT1 (min): 7.43) to give the title compound (220 mg, 60%) as a white solid. LCMS (ES, m / z): [M+H] + =297. 1H NMR (400 MHz, DMSO-d6): δ 12.87 (s, 1H), 7.83 – 7.81 (m, 2H), 7.54 –7.46 (m, 3H), 4.13 (d, J = 2.6 Hz, 2H), 3.30 (t, J = 2.6 Hz, 1H), 2. 30(s,3H).

[0804] Synthesis Method GM

[0805] 7-(trifluoromethyl)-2-thioalkyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one was synthesized using synthetic method G, and then converted to Example 342 using synthetic method M:

[0806]

[0807] LCMS (ES, m / z): [M+H] + = 415. 1H NMR (400 MHz, DMSO-d6) δ 7.43 – 7.39(m, 2H), 6.45 (tt, J = 53.7, 3.0 Hz, 1H), 4.69 (td, J = 15.2, 3.0 Hz, 2H).

[0808] Examples of synthesizing table GM using the synthesis method GM:

[0809] Table GM

[0810]

[0811] Examples 380 and 381 - 2-ethoxy-7-[oxacyclohexane-2-yl]-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0812]

[0813] Sodium ethoxide (1.9 g, 5.6 mmol, 20 wt% in EtOH) was added to a stirred solution of 2-methanesulfonyl-7-(oxecyclohexane-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4- (800 mg, 1.87 mmol) in DMF (8 mL). The resulting mixture was stirred at room temperature for 2 h and then acidified to pH 6 with 2N HCl. The mixture was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 60%–90% gradient over 10 min; UV 254 nm, to give a racemic mixture (330 mg). SFC purification (column: CHIRALPAK IG 3*25 cm, 5 μm; mobile phase A: CO2, mobile phase B: MeOH; flow rate: 100 mL / min; isocratic strength 20% B; CT (°C): 35; pressure (bar): 100; wavelength: 274 / 224 nm; RT1 (min): 5.38; RT1 (min): 6.88; MeOH:DCM, 1:1; injection volume: 2.5 mL) yielded enantiomer 1 (140 mg) with RT1 (min): 5.38 and enantiomer 2 (140 mg) with RT1 (min): 6.88, both as yellow solids.

[0814] Enantiomer 1 (140 mg) was purified by Prep-HPLC (column: Sunfire prep C18 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 47%-67% B over 10 min; wavelength: 254 nm / 220 nm; RT1 (min): 10.53) to give the title compound (91 mg, 12%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + 395. 1 H NMR (400 MHz, DMSO-d6) δ 12.87 (s, 1H), 7.66-7.62 (m, 2H), 4.51 (dd, J 11.1, 2.2 Hz, 1H), 4.45 (q, J 7.2 Hz, 2H), 4.00 (d, J 11.1 Hz,1H), 3.59 (m, 1H), 2.08 (m, 1H), 1.97 (m, 1H), 1.76 (d, J 13.4 Hz, 1H), 1.64-1.58 (m, 3H), 1.38 (t, J 7.1 Hz, 3H).

[0815] Enantiomer 2 (140 mg) was purified by Prep-HPLC (column: Sunfire prep C18 30*150 mm, 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 47%-67% B over 10 min; wavelength: 254 nm / 220 nm; RT1 (min): 10.6) to give the title compound (98 mg, 13%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + 395. 1 H NMR (400 MHz, DMSO-d6) δ 12.87 (s, 1H), 7.67-7.63 (m, 2H), 4.54 (dd, J 11.1, 2.3 Hz, 1H), 4.46 (q, J 7.1, 2H), 4.01 (d, J 11.1 Hz, 1H), 3.59 (m, 1H), 2.06 (m, 1H), 1.97 (m, 1H), 1.77 (d, J 13.4 Hz, 1H), 1.64-1.58 (m, 3H), 1.38 (t, J 7.1 Hz, 3H).

[0816] Synthesis method AA

[0817] Example 382 - 8-[4-(difluoromethyl)phenyl]-7-(pyridin-2-yl)-2-(2,2,2-trifluoroethoxy)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0818]

[0819] Step 1: 5-(pyridin-2-yl)-2H-pyrazole-3-amine

[0820] At room temperature, hydrazine hydrate (8.56 g, 137 mmol, 80 wt%) and TFA (15.60 g) were added fractionally to a stirred solution of 3-oxo-3-(pyridin-2-yl)propionitrile (10.0 g, 68.4 mmol) in EtOH (100 mL). The resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography, eluting with DCM / MeOH (10:1), to give the title compound (10.6 g, 97%) as a yellow solid. LCMS (ES, m / z): [M+H] + 161.

[0821] Step 2: 4-Bromo-5-(pyridin-2-yl)-2H-pyrazol-3-amine

[0822] NBS (6.67 g, 37.5 mmol) was added to a stirred solution of 5-(pyridin-2-yl)-2H-pyrazol-3-amine (6.0 g, 37 mmol) in ACN (30 mL) and THF (30 mL) at room temperature. The resulting mixture was stirred at room temperature for 1 h, then concentrated under reduced pressure to give the title compound (9 g, overnight) as a brown solid. LCMS (ES, m / z): [M+H] + 239,241.

[0823] Step 3: N-{[4-bromo-5-(pyridin-2-yl)-2H-pyrazol-3-yl]carbamate}ethyl carbamate

[0824] At room temperature, ethyl N-thiocarbonylcarbamate (4.94 g, 37.6 mmol) was added to a stirred solution of 4-bromo-5-(pyridin-2-yl)-2H-pyrazole-3-amine (estimated 37 mmol) in DMF (90 mL), and the resulting mixture was stirred at room temperature for 1 h. Water (150 mL) was added, and the mixture was then extracted with EA (3 × 100 mL). The combined organic layers were washed with water (2 × 50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the title compound (11 g, 80% (2 steps)) as a yellow oil. LCMS (ES, m / z): [M+H] + 372, 374.

[0825] Step 4: 8-Bromo-7-(pyridin-2-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0826] At room temperature, EtONa (11.0 g, 32.4 mmol, 20 wt% in EtOH) was added to a stirred solution of N-{[4-bromo-5-(pyridin-2-yl)-2H-pyrazol-3-yl]aminothioyl}-carbamate (6.0 g crude, 16 mmol) in EtOH (50 mL), and the resulting mixture was stirred at 80 °C for 2 h. The mixture was concentrated under reduced pressure, redissolved in water (100 mL), acidified to pH 6 with 2N HCl, and extracted with EA (3 × 100 mL). The combined organic layers were washed with brine (2 × 100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with DCM / MeOH (8:1), to give the title compound (600 mg, 11%) as a yellow solid. LCMS (ES, m / z): [M+H] + 326, 328.

[0827] Step 5: 8-Bromo-2-(methylthioalkyl)-7-(pyridin-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0828] At room temperature, K₂CO₃ (2.05 g, 14.8 mmol) and MeI (560 mg, 3.95 mmol) were added to a stirred solution of 8-bromo-7-(pyridin-2-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (1.6 g, 4.9 mmol) in DMF (16 mL). The resulting mixture was stirred at room temperature for 1 h, then filtered, and the filter cake was washed with DMF (3 × 2 mL). The filtrate was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 40%–70% gradient over 10 min; UV 254 nm, to give the title compound (530 mg, 32%) as a yellow solid. LCMS (ES, m / z): [M+H] + 338, 340.

[0829] Step 6: 8-Bromo-2-methanesulfonyl-7-(pyridin-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one and 8-Bromo-2-(methylsulfinyl)-7-(pyridin-2-yl)pyrazolo[1,5-a][1,3,5]triazin-4(3H)-one

[0830] At 0 °C, mCPBA (429 mg, 2.48 mmol) was added to a stirred solution of 8-bromo-2-(methylthioalkyl)-7-(pyridin-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (420 mg, 1.24 mmol) in DCM (10 mL). The resulting mixture was stirred at 0 °C for 15 min and then at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 30%–70% gradient over 10 min; UV 254 nm, to give the title compound (355 mg mixture, 77%) as a yellow solid. LCMS (ES, m / z): [M+H] + 354, 356 and 370, 372.

[0831] Step 7: 8-Bromo-7-(pyridin-2-yl)-2-(2,2,2-trifluoroethoxy)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0832] NaH (32 mg, 0.81 mmol, 60% mineral oil) was added to a stirred solution of trifluoroethanol (81 mg, 0.81 mmol) in DMF (2.00 mL) at 0 °C under N2, and the resulting mixture was stirred at 0 °C for 30 min. 8-Bromo-2-methanesulfonyl-7-(pyridin-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one and 8-bromo-2-(methylsulfinyl)-7-(pyridin-2-yl)pyrazolo[1,5-a][1,3,5]triazin-4(3H)-one (100 mg mixture, 0.270 mmol) were added at 0 °C, and the resulting mixture was stirred at room temperature for 2 h. Water (10 mL) was added, and the mixture was acidified to pH 5 with 2N HCl (0.5 mL) and extracted with EA (3 × 20 mL). The combined organic layers were washed with water (2 × 10 mL) and brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the title compound (113 mg, overweight) as a yellow solid. LCMS (ES, m / z): [M+H] + 390, 392.

[0833] Step 8: 8-[4-(difluoromethyl)phenyl]-7-(pyridin-2-yl)-2-(2,2,2-trifluoroethoxy)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0834] At room temperature, K₂CO₃ (120 mg, 0.870 mmol) and Pd(dtbpf)Cl₂ (19 mg, 0.029 mmol) were added to a stirred mixture of 8-bromo-7-(pyridin-2-yl)-2-(2,2,2-trifluoroethoxy)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (113 mg, 0.290 mmol) and 4-(difluoromethyl)phenylboronic acid (99 mg, 0.58 mmol) in dioxane (2 mL) and H₂O (0.5 mL). The resulting mixture was stirred overnight at 80 °C under N₂. Water (20 mL) was added to the mixture, and the mixture was then extracted with EA (3 × 20 mL). The combined organic layers were washed with water (2 × 10 mL) and brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 30%-80% gradient over 10 min; UV 254 nm. The crude product (25 mg) was purified by Prep-HPLC (column: Xselect CSH C18 OBD column 30*150 mm 5 μm; mobile phase A: water (0.1% FA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 39%-59% over 8 min; wavelength: 254 nm / 220 nm; RT1 (min): 6.48) to give the title compound (11.6 mg, 9%) as a grayish-white solid. LCMS (ES, m / z): [M+H] + 438. 1 H NMR: (400 MHz, DMSO-d6) δ 13.31 (s, 1H), 8.54 (d, J 4.8 Hz, 1H), 7.95 (m, 1H), 7.85 (d, J7.8 Hz, 1H), 7.55-7.50 (m, 4H), 7.46 (dd, J 7.6, 4.9 Hz, 1H), 7.03 (t, J 56.1Hz, 1H), 5.10 (q, J 8.8 Hz, 2H).

[0835] The embodiments in Table AA were synthesized using synthesis method AA.

[0836] Table AA

[0837]

[0838] Example 385 - 8-[4-(difluoromethyl)phenyl]-2-(prop-2-yn-1-ylthioalkyl)-7-(pyridin-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0839]

[0840] Step 1: 8-(4-bromophenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-(pyridin-2-yl)pyrazolo[1,5-a][1,3,5]triazin-4-one

[0841] At room temperature, K₂CO₃ (331 mg, 2.40 mmol) and 4-methoxybenzyl chloride (376 mg, 2.40 mmol) were added to a stirred solution of 8-(4-bromophenyl)-7-(pyridin-2-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (synthesized using synthetic method A; 320 mg, 0.800 mmol) in DMF (6 mL), and the mixture was stirred overnight at 50 °C. The mixture was cooled, filtered, and the filter cake was washed with DMF (3 × 2 mL). The filtrate was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 50%–100% gradient over 10 min; UV 254 nm, to give the title compound (340 mg, 66%) as a yellow solid. LCMS (ES, m / z): [M+H] + 640, 642.

[0842] Step 2: 8-[4-(difluoromethyl)phenyl]-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-(pyridin-2-yl)pyrazolo[1,5-a][1,3,5]triazine-4-one

[0843] At room temperature, 2-[2-[2-(diphenylphosphine)phenoxy]phenyl}diphenylphosphine (CAS: 166330-10-5; 27 mg, 0.050 mmol) and Pd(dba)2 (29 mg, 0.050 mmol) were added to a stirred mixture of 8-(4-bromophenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-(pyridin-2-yl)pyrazolo[1,5-a][1,3,5]triazine-4-one (320 mg, 0.500 mmol) and [1,3-bis[2,6-bis(isopropyl)phenyl]-2-imidazoalkyl]difluoromethylsilver(I) (CAS: 1643366-13-5; 270 mg, 0.750 mmol) in toluene (6 mL) and stirred mixture of 8-(4-bromophenyl)-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-(pyridin-2-yl)pyrazolo[1,5-a][1,3,5]triazine-4-one (320 mg, 0.500 mmol) and [1,3-bis[2,6-bis(isopropyl)phenyl]-2-imidazoalkyl]difluoromethylsilver(I) (CAS: 1643366-13-5; 27 mg, 0.750 mmol) in toluene (6 mL). The resulting mixture was stirred overnight at 80 °C under N2. The cooled mixture was concentrated under reduced pressure, suspended in water (20 mL), and extracted with EA (3 × 20 mL). The combined organic layers were washed with brine (2 × 10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 40%–100% gradient over 10 min; UV 254 nm, to give the title compound (134 mg, 44%) as a yellow solid. LCMS (ES, m / z): [M+H] + 612.

[0844] Step 3: 8-[4-(difluoromethyl)phenyl]-7-(pyridin-2-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0845] A solution of 8-[4-(difluoromethyl)phenyl]-3-[(4-methoxyphenyl)methyl]-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-(pyridin-2-yl)pyrazolo[1,5-a][1,3,5]triazin-4-one (113 mg, 0.185 mmol) in TFA (2 mL) was stirred at 80 °C for 2 h, and then the cooled mixture was concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 30%–80% gradient over 10 min; UV 254 nm, to give the title compound (68 mg, 99%) as a yellow solid. LCMS (ES, m / z): [M+H] + 372.

[0846] Step 4: 8-[4-(difluoromethyl)phenyl]-2-(prop-2-yn-1-ylthioalkyl)-7-(pyridin-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0847] At room temperature, K₂CO₃ (78 mg, 0.56 mmol) and propargyl bromide (16 mg, 0.13 mmol) were added to a stirred solution of 8-[4-(difluoromethyl)phenyl]-7-(pyridin-2-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (70 mg, 0.19 mmol) in DMF (2 mL). The resulting mixture was stirred at room temperature for 1 h, and then water (20 mL) was added. The mixture was extracted with EA (3 x 20 mL), and the combined organic layers were washed with water (2 x 10 mL) and brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 30%–80% gradient over 10 min; UV 254 nm. The crude product (40 mg) was purified by Prep-HPLC (column: Xselect CSH C18 OBD column 30*150 mm, 5 μm; mobile phase A: water (0.1% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 35%-50% B over 8 min; wavelength: 254 nm / 220 nm; RT1 (min): 8.58) to give the title compound (26 mg, 34%) as a white solid. LCMS (ES, m / z): [M+H] + 410. 1 H NMR: (400 MHz, DMSO-d6) δ 13.24 (s, 1H), 8.56 (d, J 4.7 Hz, 1H), 7.95 (m, 1H), 7.86 (d, J7.8 Hz, 1H), 7.64 (d, J 8.0 Hz, 2H), 7.51-7.45 (m, 3H), 7.03 (t, J 56.1 Hz, 1H), 4.03 (d, J 2.7 Hz, 2H), 3.31 (t, J 2.6 Hz, 1H).

[0848] Example 386-2-(prop-2-yn-1-yloxy)-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0849]

[0850] Step 1: 8-Bromo-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-(pyrimidin-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0851] At room temperature, 4-methoxybenzyl chloride (2.60 g, 16.6 mmol) and K₂CO₃ (2.30 g, 16.6 mmol) were added to a stirred solution of 8-bromo-7-(pyrimidin-2-yl)-2-thioalkyl-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (synthesized using synthetic method AA, 1.80 g, 5.54 mmol) in DMF (20 mL), and the mixture was stirred overnight at room temperature. The mixture was diluted with water (40 mL) and extracted with EA (3 × 20 mL). The combined organic layers were washed with water (2 × 20 mL) and brine (20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA) in a 40%–60% gradient over 10 min; UV 254 nm, to give the title compound (2.1 g, 85%) as a yellow solid. LCMS (ES, m / z): [M+H] + 445, 447.

[0852] Step 2: 2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0853] At room temperature under N2, Pd(dtbpf)Cl2 (0.29 g, 0.45 mmol) and K2CO3 (1.24 g, 8.98 mmol) were added to a stirred mixture of 8-bromo-2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-(pyrimidin-2-yl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (2.0 g, 4.5 mmol) and 3,4,5-trifluorophenylboronic acid (3.95 g, 22.5 mmol) in dioxane (20 mL) and H2O (2 mL). The resulting mixture was stirred overnight at 85 °C under N2. The cooled mixture was diluted with water (40 mL) and extracted with EA (3 × 20 mL). The combined organic layers were washed with brine (2 × 20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA) in a 60%–80% gradient over 10 min; UV 254 nm, yielding the title compound (240 mg, 11%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + 497.

[0854] Step 3: 7-(pyrimidin-2-yl)-2-thioalkyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0855] A solution of 2-{[(4-methoxyphenyl)methyl]thioalkyl}-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (240 mg, 0.483 mmol) in TFA (4 mL) was stirred for 2 h at 70 °C. The cooled mixture was concentrated under reduced pressure, and the residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 40%–60% gradient over 10 min; UV 254 nm, to give the title compound (80 mg, 44%) as a pale yellow solid. LCMS (ES, m / z): [M+H] + 377.

[0856] Step 4: 2-(methylthioalkyl)-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0857] MeI (15 mg, 0.11 mmol) and K₂CO₃ (59 mg, 0.43 mmol) were added to a stirred solution of 7-(pyrimidin-2-yl)-2-thioalkyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (80 mg, 0.21 mmol) in DMF (2 mL), and the resulting mixture was stirred at room temperature for 1 h. The mixture was filtered and the filter cake was washed with DMF (2 × 1 mL). The filtrate was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 40%–50% gradient over 10 min; UV 254 nm, to give the title compound (40 mg, 48%) as a white solid. LCMS (ES, m / z): [M+H] + 391.

[0858] Step 5: 2-Methanesulfonyl-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one and 2-methanesulfonyl-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0859] At room temperature, mCPBA (44 mg, 0.26 mmol) was added to a stirred solution of 2-(methylthioalkyl)-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (50 mg, 0.13 mmol) in DCM (2 mL). The resulting mixture was stirred overnight at room temperature, then filtered, and the filter cake was washed with DCM (2 × 1 mL). The filtrate was concentrated under reduced pressure, and the residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 40%–70% gradient over 10 min; UV 254 nm, to give the title compound (30 mg mixture, 71%) as a white solid. LCMS (ES, m / z): [MH] - 405 and 421.

[0860] Step 6: 2-(prop-2-yn-1-yloxy)-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0861] At 0 °C, NaH (3.9 mg, 0.098 mmol, 60% mineral oil) was added to a stirred solution of propargyl alcohol (5.5 mg, 0.098 mmol) in DMF (1 mL). The resulting mixture was stirred at 0 °C for 30 min. 2-Methylsulfinyl-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one and 2-methanesulfonyl-7-(pyrimidin-2-yl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (30 mg mixture, 0.049 mmol) were added at 0 °C, and the mixture was then stirred at room temperature for 1 h. The mixture was neutralized to pH 7 with 2M HCl (0.3 mL) and then purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 40%–50% gradient over 10 min; UV 254 nm. The crude product was purified by Prep-HPLC (column: Sunfire Prep C18 column 30*150 mm, 5 μm; mobile phase A: water (0.1% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 32%–50% B over 10 min; wavelength: 254 nm / 220 nm; RT1 (min): 10.05) to give the title compound (9 mg, 46%) as a white solid. LCMS (ES, m / z): [M+H] + 399. 1 H NMR: (300 MHz, DMSO-d6) δ 13.25 (s, 1H), 8.94-8.93 (m,2H), 7.61-7.59 (m, 1H), 7.33 (t, J 8.4 Hz, 2H), 5.11 (d, J 2.2 Hz, 2H), 3.76(t, J 2.2 Hz, 1H).

[0862] Synthesis method BB

[0863] Example 867 - 7-[1-(benzyloxy)ethyl]-2-ethoxy-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0864]

[0865] Step 1: 4-(benzyloxy)-3-oxo-2-(3,4,5-trifluorophenyl)pentanilonitrile

[0866] At 0 °C, NaH (1.40 g, 35.1 mmol, 60% mineral oil) was added to a stirred solution of 2-(3,4,5-trifluorophenyl)acetonitrile (2.0 g, 12 mmol) in THF (30 mL). The mixture was stirred at 0 °C for 30 min, and then methyl (2S)-2-(benzyloxy)propionate (2.72 g, 14.0 mmol) was added at 0 °C. The resulting mixture was stirred overnight at room temperature. The mixture was acidified to pH 5 with 2 M HCl at 0 °C, diluted with water (50 mL), and extracted with EA (3 × 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the title compound (4.08 g, overweight) as a yellow solid. LCMS (ES, m / z): [MH] - 332.

[0867] Step 2: 3-(1-(benzyloxy)ethyl)-4-(3,4,5-trifluorophenyl)-1H-pyrazole-5-amine

[0868] At 0 °C, N₂H₄·H₂O (1.61 g, 25.7 mmol, 80 wt%) and TFA (5.9 g, 51 mmol) were added to a stirred solution of 4-(benzyloxy)-3-oxo-2-(3,4,5-trifluorophenyl)pentanonitrile (assumed to be 12 mmol) in EtOH (40 mL), and the mixture was stirred at 80 °C for 4 h. The cooled solution was diluted with water (80 mL) and extracted with EA (3 × 80 mL). The combined organic layers were washed with brine (2 × 80 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to give the racemic title compound (4.91 g, overweight) as a yellow solid. LCMS (ES, m / z): [M+H] + 348.

[0869] Step 3: 1-(3-(1-(benzyloxy)ethyl)-4-(3,4,5-trifluorophenyl)-1H-pyrazol-5-yl)-3-(ethoxymethyl)thiourea

[0870] At room temperature under N2, N-thiocarbonylcarbamate (3.71 g, 28.3 mmol) was added to a stirred solution of 3-(1-(benzyloxy)ethyl)-4-(3,4,5-trifluorophenyl)-1H-pyrazole-5-amine (assumed to be 12 mmol) in DMF (50 mL). The resulting mixture was stirred at room temperature for 1 h, then diluted with water (100 mL) and extracted with EA (3 × 100 mL). The combined organic layers were washed with water (2 × 100 mL) and brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (6.84 g, overweight) as a yellow oil. LCMS (ES, m / z): [M+H] + 479.

[0871] Step 4: 7-(1-(benzyloxy)ethyl)-2-mercapto-8-(3,4,5-trifluorophenyl)pyrazolo[1,5-a][1,3,5]triazine-4(3H)-one

[0872] A solution of ethyl 1-(3-(1-(benzyloxy)ethyl)-4-(3,4,5-trifluorophenyl)-1H-pyrazol-5-yl)-3-(ethoxymethyl)thiourea (6.84 g, 14.3 mmol) in 2 N NaOH (70 mL) was stirred at 50 °C for 1 h. The mixture was acidified to pH 6 with 2 M HCl (75 mL) at 0 °C, diluted with water (150 mL), and extracted with EA (3 × 100 mL). The combined organic layers were washed with brine (2 × 100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 30%–80% gradient over 60 min; UV 254 nm, to give the title compound (1.32 g, 25% by 4 steps) as a pale yellow solid. LCMS (ES, m / z): [M+H] + 433.

[0873] Step 5: 7-(1-(benzyloxy)ethyl)-2-(methylthio)-8-(3,4,5-trifluorophenyl)pyrazolo[1,5-a][1,3,5]triazine-4(3H)-one

[0874] At room temperature, K₂CO₃ (1.22 g, 8.81 mmol) and MeI (0.67 g, 4.7 mmol) were added to a stirred solution of 7-(1-(benzyloxy)ethyl)-2-mercapto-8-(3,4,5-trifluorophenyl)pyrazolo[1,5-a][1,3,5]triazine-4(3H)-one (1.27 g, 2.94 mmol) in DMF (15 mL). The resulting mixture was stirred at room temperature for 1 h, then acidified to pH 6 with 2M HCl (10 mL) at 0 °C, diluted with water (25 mL), and extracted with EA (3 × 25 mL). The combined organic layers were washed with water (2 × 25 mL) and brine (25 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.05% FA) in a 30%–80% gradient over 30 min; UV 254 nm, to give the title compound (891 mg, 68%) as a white solid. LCMS (ES, m / z): [M+H] + 447.

[0875] Step 6: 7-[1-(benzyloxy)ethyl]-2-methanesulfinyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0876] At room temperature, a solution of oxone (6.61 g, 10.8 mmol) in 20 mL H₂O was added to a stirred solution of 7-[(1S)-1-(benzyloxy)ethyl]-2-(methylthioalkyl)-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one (4.0 g, 9.0 mmol) in AcOH (80 mL). The resulting mixture was stirred at room temperature for 1 h, then diluted with water (100 mL) and extracted with EA (2 × 70 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EA (1:1), to give the title compound (1.22 g, 29%) as a brown oil. LCMS (ES, m / z): [M+H] + 463.

[0877] Step 7: 7-[1-(benzyloxy)ethyl]-2-ethoxy-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazine-4-one

[0878] At room temperature, a mixture of 7-[1-(benzyloxy)ethyl]-2-methylsulfinyl-8-(3,4,5-trifluorophenyl)-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one (100 mg, 0.216 mmol) and EtONa (220 mg, 0.648 mmol, 20 wt% in EtOH) in DMF (3 mL) was stirred for 1 h. The resulting mixture was purified by reversed-phase rapid chromatography: C18 silica gel; ACN aqueous solution (0.1% FA), 40%–80% gradient over 10 min; UV 254 nm, to give the crude product. The crude product (80 mg) was purified by Pr...

Claims

1. A compound of formula (I) or formula (II) or its pharmaceutically acceptable salt in R 1 It is hydrogen or an optionally substituted C1-C3 alkyl group; R 2 It is -SL 2 -R 2a or -OL 2 -R 2a ; L 2 It is a C1-C3 alkylene group with a bond or optional substitution; R 2a Choose from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, and optionally substituted 5-10-membered heteroaryl, wherein R 2a R 2b Replace n examples; Each R 2b Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or R 2b Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups; R 3 Choose from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 5-10-membered heteroaryl, wherein R 3 R 3a Replace m examples; Each R 3a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or R 3a Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups; R 4 Choose from the following groups: hydrogen, halogen, -CN, -OH, -OR c -(CH2) 1-3 R c Optionally substituted C2-C6 alkyl groups, optionally substituted C3-C6 carbocyclic groups, optionally substituted 3-10-membered heterocyclic groups, optionally substituted phenyl groups, and optionally substituted 5-10-membered heteroaryl groups, wherein R 4 R 4a Replace p examples; Each R 4a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or R 4a Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups; Each R c Independently selected from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or R c Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups; n is 0, 1, 2, 3, 4, or 5; and m is 0, 1, 2, 3, 4, or 5; and p is 0, 1, 2, 3, 4 or 5.

2. A compound of formula (I) or formula (II) or its pharmaceutically acceptable salt in R 1 It is hydrogen or an optionally substituted C1-C3 alkyl group; R 2 It is -SL 2 -R 2a or -OL 2 -R 2a ; L 2 It is a C1-C3 alkylene group with a bond or optional substitution; R 2a The group consisting of: C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, and optionally substituted 5-6-membered heteroaryl, wherein R 2a R 2b Replace n examples; Each R 2b Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl groups, optionally substituted C2-C6 alkenyl groups, optionally substituted C2-C6 ynyl groups, optionally substituted 3-7 membered carbocyclic groups, and optionally substituted 3-10 membered heterocyclic groups, or R 2b Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups; R 3 Choose from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 5-10-membered heteroaryl, wherein R 3 R 3a Replace m examples; Each R 3a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or R 3a Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups; R 4 Choose from the following groups: hydrogen, halogen, -CN, -OH, -OR c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl groups, optionally substituted C3-C6 carbocyclic groups, optionally substituted 3-10-membered heterocyclic groups, optionally substituted phenyl groups, and optionally substituted 5-10-membered heteroaryl groups, wherein R 4 R 4a Replace p examples; Each R 4a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or R 4a Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups; Each R c Independently selected from the group consisting of: optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl and optionally substituted 5-10-membered heteroaryl, or R c Two examples can be optionally combined with their intermediary atoms to form optionally substituted 4-7 membered heterocyclic groups; n is 0, 1, 2, 3, 4, or 5; and m is 0, 1, 2, 3, 4, or 5; and p is 0, 1, 2, 3, 4, or 5 Where R 2 When it is -S-CH2-pyridyl or -S-CH(CH3)-phenyl, R 4 It is not methyl.

3. The compound of claim 1, wherein R 2 It is -SL 2 -R 2a .

4. The compound of claim 1, wherein R 2 Is -OL 2 -R 2a .

5. The compound according to any one of claims 1-4, wherein L 2 It is a key.

6. The compound according to any one of claims 1-4, wherein L 2 It is -CH2-.

7. The compound according to any one of claims 1-6, wherein R 2a Choose from the group consisting of: optionally substituted C1-C3 alkyl, optionally substituted C2-C3 alkenyl, optionally substituted C2-C4 ynyl, optionally substituted 3-6 membered carbocyclic, optionally substituted phenyl, and optionally substituted 5-6 membered heteroaryl, wherein R 2a R 2b Replace n examples.

8. The compound of claim 7, wherein R 2a It is an optionally substituted C1-C3 alkyl group, wherein R 2a R 2b Replace n examples.

9. The compound of claim 8, wherein R a Choose from the group consisting of: optionally substituted methyl, optionally substituted ethyl, and optionally n-propyl.

10. The compound of claim 7, wherein R 2a It is an optional substituted C2-C3 alkenyl group, wherein R 2a R 2b Replace n examples.

11. The compound of claim 10, wherein R 2a It is an allyl group with optional substitution.

12. The compound of claim 7, wherein R 2a It is an optional substituted C2-C4 ynyl group, wherein R 2a R 2b Replace n examples.

13. The compound of claim 12, wherein R 2a It is an optional substituted propynyl group.

14. The compound of claim 7, wherein R 2a It is an optional substituted 3-6 membered carbon cyclogroup, wherein R 2a R 2b Replace n examples.

15. The compound of claim 14, wherein R 2a It is an optionally substituted cyclopropyl group or an optionally substituted cyclopropyl group.

16. The compound of claim 1, R 2 Choose from the following groups: and .

17. The compound according to any one of claims 1-5, wherein R 3 It is an optionally substituted phenyl or an optionally substituted 6-membered heteroaryl group.

18. The compound of claim 17, wherein R 3 It is an optionally substituted phenyl or optionally substituted pyridyl group.

19. The compound according to any one of claims 1-5, wherein R 3 It is an optional substituted 3-7 membered carbon cyclogroup.

20. The compound according to any one of claims 1-5, wherein R 3 It is an optional substituted 3-7 membered heterocyclic group.

21. The compound according to any one of claims 1-17, wherein the compound has formula (Ia) or formula (II-a). Or its pharmaceutically acceptable salt.

22. The compound according to any one of claims 1-21, wherein R 1 It is hydrogen.

23. The compound according to any one of claims 1-2221, wherein each R 3a Choose independently from the following groups: halogen, -OH, -OR c , -NH2, -CN, -NO2, -C(O)OH, -C(O)OR c -C(O)R c -NHR c -N(R) c )2、-C(O)NHR c -C(O)N(R) c )2、-S(O)2R c -(CH2) 1-3 R c Optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 ynyl, optionally substituted 3-7-membered carbocyclic, optionally substituted 3-10-membered heterocyclic, optionally substituted phenyl, and optionally substituted 5-10-membered heteroaryl.

24. The compound of claim 23, wherein R 3a It is halogen.

25. The compound of claim 24, wherein R 3a It is a chlorinated or fluorinated group.

26. The compound of claim 23, wherein R 3a Yes - OR c .

27. The compound of claim 26, wherein R 3a Yes-OMe.

28. The compound of claim 26, wherein R c It is an optional substituted 5-6 membered heterocyclic group.

29. The compound of claim 23, wherein R 3a It is an optional substituted C1-C6 alkyl group.

30. The compound of claim 29, wherein R 3a It is an optional substituted methyl group.

31. The compound of claim 23, wherein R 3a It is -(CH2) 1-3 R c .

32. The compound of claim 31, wherein R c It is an optional 6-membered heterocyclic group.

33. The compound of claim 23, wherein R 3a Yes-S(O)2R c .

34. The compound of claim 33, wherein R c It is an optional substituted C1-C3 alkyl group.

35. The compound of claim 23, wherein R 3a It is -C(O)N(R) c )2.

36. The compound of claim 23, wherein R 3a Yes - CN.

37. The compound according to any one of claims 1-22, wherein m is 2, and R c The two examples, together with their intercalary atoms, form 4-7 membered heterocyclic groups with optional substitution.

38. The compound of claim 37, wherein R c The two examples, together with their intermediary atoms, form optional substituted 5-membered heterocyclic groups.

39. The compound according to any one of claims 1-23, wherein R 4 Choose from the following groups: halogen, -CN, -OH, -OR c -CH2R c Optionally substituted C2-C4 alkyl groups, optionally substituted C3-C4 carbocyclic groups, optionally substituted 3-7-membered heterocyclic groups, optionally substituted phenyl groups, and optionally substituted 5-6-membered heteroaryl groups, wherein R 4 R 4a Replace p examples.

40. The compound according to any one of claims 1-23, wherein R 4 It is an optional substituted C1-C4 alkyl group.

41. The compound of claim 40, wherein R 4 It is an optional substituted methyl group.

42. The compound as claimed in claim 39, wherein R 4 It is an optional substituted C2-C4 alkyl group.

43. The compound of claim 42, wherein R 4 It is optionally substituted ethyl, optionally substituted isopropyl, optionally substituted tert-butyl, and optionally substituted sec-butyl.

44. The compound of claim 39, wherein R 4 It is an optional substituted 3-7 membered heterocyclic group.

45. The compound of claim 44, wherein R 4 It is an optional substituted 5-7 membered heterocyclic group.

46. ​​The compound of claim 44, wherein R 4 Choose from the group consisting of: optionally substituted tetrahydrofuranyl, optionally substituted tetrahydropyranyl, optionally substituted piperidinyl, optionally substituted morpholinyl and optionally substituted oxazine-heptyl.

47. The compound of claim 39, wherein R 4 It is an optional substituted phenyl group.

48. The compound of claim 39, wherein R 4 It is an optional substituted 5-6 member heteroaryl group.

49. The compound of claim 48, wherein R 4 Choose from the group consisting of: optionally substituted imidazolyl, optionally substituted pyridyl, optionally substituted pyrimidinyl, and optionally substituted pyrazinyl.

50. The compound according to any one of claims 1-23, wherein R 4 Choose from the following groups: hydrogen, methyl, and .

51. A compound selected from the group consisting of those compounds found in Table 1, or a pharmaceutically acceptable salt thereof.

52. A pharmaceutical composition comprising the compound of any one of claims 1-51 and a pharmaceutically acceptable adjuvant.

53. A method of treating or preventing a disease or disorder associated with protein misfolding, comprising administering to a subject in need a therapeutically effective amount of any one of claims 1-51 or the pharmaceutical composition of claim 52.

54. The method of claim 53, wherein the protein misfolding disease is Parkinson's disease.

55. The method of claim 53, wherein the protein misfolding disease or disorder is selected from the group consisting of: Parkinson's disease, Lewy body dementia, or multiple system atrophy.

56. The method of claim 53, wherein the protein misfolding disease or disorder is selected from the group consisting of: synucleinosis, amyloidosis, tau disease, prions, Creutzfeldt-Jakob disease, spongiform encephalopathy, neurodegenerative diseases, Down syndrome, or cystic fibrosis.

57. The method of claim 53, wherein the disease or disorder associated with protein misfolding is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, Lewy body dementia, diffuse Lewy body disease, sporadic Alzheimer's disease, familial Alzheimer's disease, Lewy body variant of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, pure autonomic failure, or neurodegeneration combined with brain iron accumulation type 1.

58. The method of claim 53, wherein the disease or disorder associated with protein misfolding is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, dementia associated with Lewy body, diffuse Lewy body disease, sporadic Alzheimer's disease, familial Alzheimer's disease, Lewy body variant of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, Hallervorden-Spatz disease, pure autonomic failure, or neurodegeneration associated with type 1 brain iron accumulation.

59. The method of claim 52, wherein the disease or disorder associated with protein misfolding is Lewy body disease, Parkinson's disease, Parkinson's dementia, dementia associated with Lewy bodies, diffuse Lewy body disease, sporadic and familial Alzheimer's disease, Lewy body variants of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, Hallervorden-Spatz disease, pure autonomic failure, neurodegeneration associated with brain iron accumulation type 1, generalized neurodegenerative diseases, synucleinopathy, pure autonomic failure, dementia, tau proteinopathy, argyrophilic granuloma, Pick's disease, Down syndrome, chronic leukemia, lymphoma, schizophrenia, psychosis, or Creutzfeldt-Jakob disease.

60. A method of treating or preventing a disease or disorder associated with neurodegeneration, comprising administering to a subject in need a therapeutically effective amount of any one of claims 1-51 or the pharmaceutical composition of claim 52.

61. The method of claim 60, wherein the disease or disorder associated with neurodegeneration is Parkinson's disease.

62. The method of claim 60, wherein the disease or disorder associated with neurodegeneration is selected from the group consisting of: synucleinosis, amyloidosis, tau disease, prions, Creutzfeldt-Jakob disease, spongiform encephalopathy, Down syndrome, or cystic fibrosis.

63. The method of claim 60, wherein the disease or disorder associated with neurodegeneration is selected from Parkinson's disease, Lewy body dementia, or multiple system atrophy.

64. The method of claim 60, wherein the disease or disorder associated with neurodegeneration is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, Lewy body dementia, diffuse Lewy body disease, sporadic Alzheimer's disease, familial Alzheimer's disease, Lewy body variant of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, pure autonomic failure, or neurodegeneration combined with brain iron accumulation type 1.

65. The method of claim 60, wherein the disease or disorder associated with neurodegeneration is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, dementia combined with Lewy body, diffuse Lewy body disease, sporadic Alzheimer's disease, familial Alzheimer's disease, Lewy body variant of Alzheimer's disease, combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, Hallervorden-Spatz disease, pure autonomic failure, or neurodegeneration combined with type 1 brain iron accumulation.

66. The method of claim 60, wherein the disease or disorder associated with neurodegeneration is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, dementia associated with Lewy body, diffuse Lewy body disease, sporadic and familial Alzheimer's disease, Lewy body variants of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, Hallervorden-Spatz disease, pure autonomic failure, neurodegeneration associated with brain iron accumulation type 1, synucleinopathy, pure autonomic failure, dementia, tau proteinopathy, arginophilic granuloma, Pick's disease, Down syndrome, chronic leukemia, lymphoma, schizophrenia, psychosis, or Creutzfeldt-Jakob disease.

67. A method of treating a disease or disorder associated with α-synuclein misfolding, comprising administering to a subject in need a therapeutically effective amount of any one of claims 1-51 or the pharmaceutical composition of claim 52.

68. The method of claim 67, wherein the disease or disorder associated with α-synuclein misfolding is Parkinson's disease.

69. The method of claim 67, wherein the disease or disorder associated with α-synuclein misfolding is selected from the group consisting of: synucleinosis, amyloidosis, tau disease, prions, Creutzfeldt-Jakob disease, spongiform encephalopathy, neurodegenerative diseases, Down syndrome, or cystic fibrosis.

70. The method of claim 67, wherein the disease or disorder associated with α-synuclein misfolding is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, Lewy body dementia, diffuse Lewy body disease, sporadic Alzheimer's disease, familial Alzheimer's disease, Lewy body variant of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, pure autonomic failure, or neurodegeneration combined with brain iron accumulation type 1.

71. The method of claim 67, wherein the disease or disorder associated with α-synuclein misfolding is selected from the group consisting of: Parkinson's disease, Lewy body dementia, or multiple system atrophy.

72. The method of claim 67, wherein the disease or disorder associated with α-synuclein misfolding is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, dementia associated with Lewy body, diffuse Lewy body disease, sporadic Alzheimer's disease, familial Alzheimer's disease, Lewy body variant of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, Hallervorden-Spatz disease, pure autonomic failure, or neurodegeneration associated with type 1 brain iron accumulation.

73. The method of claim 67, wherein the disease or disorder associated with α-synuclein misfolding is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, dementia associated with Lewy bodies, diffuse Lewy body disease, sporadic and familial Alzheimer's disease, Lewy body variants of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, Hallervorden-Spatz disease, pure autonomic failure, neurodegeneration associated with brain iron accumulation type 1, generalized neurodegenerative diseases, synucleinopathy, pure autonomic failure, dementia, tau proteinopathy, auricolic granuloma, Pick's disease, Down syndrome, chronic leukemia, lymphoma, schizophrenia, psychosis, or Creutzfeldt-Jakob disease.

74. A method of treating or preventing a disease or ailment associated with the formation, deposition, accumulation, or persistence of α-synuclein oligomers, fibrils, fibers, or aggregates, comprising administering to a subject in need a therapeutically effective amount of any one of claims 1-51 or the pharmaceutical composition of claim 52.

75. The method of claim 74, wherein the disease or disorder associated with the formation, deposition, accumulation, or persistence of α-synuclein oligomers, fibrils, fibers, or aggregates is Parkinson's disease.

76. The method of claim 74, wherein the disease or disorder associated with the formation, deposition, accumulation, or persistence of α-synuclein oligomers, fibrils, fibers, or aggregates is selected from the group consisting of: Parkinson's disease, Lewy body dementia, or multiple system atrophy.

77. The method of claim 74, wherein the disease or disorder associated with the formation, deposition, accumulation, or persistence of α-synuclein oligomers, fibrils, fibrous tissues, or aggregates is selected from the group consisting of: synucleinosis, amyloidosis, tau disease, prions, Creutzfeldt-Jakob disease, spongiform encephalopathy, neurodegenerative diseases, Down syndrome, or cystic fibrosis.

78. The method of claim 74, wherein the disease or disorder associated with the formation, deposition, accumulation, or persistence of α-synuclein oligomers, fibrils, fibers, or aggregates is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, Lewy body dementia, diffuse Lewy body disease, sporadic Alzheimer's disease, familial Alzheimer's disease, Lewy body variant of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, pure autonomic failure, or neurodegeneration combined with brain iron accumulation type 1.

79. The method of claim 74, wherein the disease or disorder associated with the formation, deposition, accumulation, or persistence of α-synuclein oligomers, fibrils, fibers, or aggregates is selected from the group consisting of: Lewy body disease, Parkinson's disease, Parkinson's dementia, dementia associated with Lewy bodies, diffuse Lewy body disease, sporadic Alzheimer's disease, familial Alzheimer's disease, Lewy body variant of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontotemporal dementia, frontotemporal degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, Hallervorden-Spatz disease, pure autonomic failure, or neurodegeneration associated with type 1 brain iron accumulation.

80. The method of claim 74, wherein the disease or disorder associated with the formation, deposition, accumulation, or persistence of α-synuclein oligomers, fibrils, fibers, or aggregates is selected from the group consisting of: Parkinson's disease, Parkinson's dementia, dementia associated with Lewy bodies, diffuse Lewy body disease, sporadic and familial Alzheimer's disease, Lewy body variants of Alzheimer's disease, a combination of Alzheimer's disease and Parkinson's disease, multiple system atrophy or Shy-Drager syndrome, frontal lobe disease, etc. Temporal lobe dementia, frontotemporal lobe degeneration, familial dementia, progressive supranuclear palsy, Gaucher disease, juvenile-onset systemic axonal dystrophy, Hallervorden-Spatz disease, pure autonomic failure, neurodegeneration combined with brain iron accumulation type 1, generalized neurodegenerative diseases, synucleinopathy, pure autonomic failure, dementia, tau proteinopathy, argyrophilic granuloma, Pick's disease, Down syndrome, chronic leukemia, lymphoma, schizophrenia, psychosis, or Creutzfeldt-Jakob disease.

Images