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HK1189221B - 5,6-dihydro-imidazo[1,2-a] pyrazin-8-ylamine derivatives useful as inhibitors of beta-secretase (bace) - Google Patents

5,6-dihydro-imidazo[1,2-a] pyrazin-8-ylamine derivatives useful as inhibitors of beta-secretase (bace) Download PDF

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Publication number
HK1189221B
HK1189221B HK14102157.9A HK14102157A HK1189221B HK 1189221 B HK1189221 B HK 1189221B HK 14102157 A HK14102157 A HK 14102157A HK 1189221 B HK1189221 B HK 1189221B
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Hong Kong
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methyl
mmol
radical
group
fluorophenyl
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HK14102157.9A
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Chinese (zh)
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HK1189221A1 (en
Inventor
Andrés Avelino TRABANCO-SUÁREZ
Francisca DELGADO-JIMÉNEZ
Juan Antonio Vega Ramiro
Gary John Tresadern
Henricus Jacobus Maria Gijsen
Daniel Oehlrich
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Janssen Pharmaceutica Nv
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Priority claimed from PCT/EP2011/073522 external-priority patent/WO2012085038A1/en
Publication of HK1189221A1 publication Critical patent/HK1189221A1/en
Publication of HK1189221B publication Critical patent/HK1189221B/en

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Description

As beta - Secretase enzymes (BACE) Of inhibitors 5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -8- Amine derivatives
Technical Field
The present invention relates to novel 5, 6-dihydro-imidazo [1,2-a ] pyrazin-8-ylamine derivatives which are inhibitors of β -secretase, also known as β -site amyloid cleaving enzyme, BACE1, Asp2 or memapsin 2. The invention is also directed to pharmaceutical compositions comprising such compounds, to processes for preparing such compounds and compositions, and to the use of such compounds and compositions in the prevention and treatment of disorders in which beta-secretase is involved, such as Alzheimer's Disease (AD), mild cognitive impairment, senility, dementia with lewy bodies, cerebral amyloid angiopathy, multi-infarct dementia, down's syndrome, dementia associated with stroke, dementia associated with parkinson's disease or dementia associated with beta-amyloid.
Background
Alzheimer's Disease (AD) is a neurodegenerative disease associated with aging. AD patients suffer from cognitive deficits and memory loss as well as behavioral problems such as anxiety. More than 90% of AD patients suffer from sporadic forms of the disorder, while less than 10% of cases are familial or hereditary. In the united states, about 10 people aged 65 have AD in 1 out of 85, and 1 out of every 2 individuals. The average life expectancy from the initial diagnosis is 7-10 years, and AD patients need to be specifically cared for in extremely expensive assisted living facilities or by family members. As the number of elderly in the population increases, medical attention to AD is increasing. Currently available treatments for AD only treat the symptoms of the disease and include acetylcholinesterase inhibitors to improve cognitive ability and anxiolytics and antipsychotics to control behavioral problems associated with the disease.
A pathological hallmark in the brain of AD patients is neuronal fibrillary tangles, which are generated by hyperphosphorylation of amyloid plaques resulting from the aggregation of tau protein and beta-amyloid 1-42 (a β 1-42) peptide. A β 1-42 forms oligomers, then fibrils, and finally amyloid plaques. It is believed that the oligomers and fibrils are particularly neurotoxic and may cause most of the neurological damage associated with AD. Drugs that prevent the formation of a β 1-42 may be drugs that alleviate AD. A β 1-42 is produced from Amyloid Precursor Protein (APP) which comprises 770 amino acids. The N-terminus of A.beta.1-42 is cleaved by beta-secretase (BACE) and then gamma-secretase cleaves the C-terminus. In addition to A.beta.1-42, gamma-secretase also releases the main cleavage products A.beta.1-40 as well as A.beta.1-38 and A.beta.1-43. These a β forms can also aggregate to form oligomers and fibrils. Therefore, inhibitors of BACE are expected to prevent the formation of A β 1-42 and A β 1-40, A β 1-38 and A β 1-43 and would be potential therapeutic agents for the treatment of AD.
Summary of The Invention
The invention relates to a compound of formula (I)
Or a tautomer or stereoisomeric form thereof, wherein
R1And R2Independently selected from hydrogen, halo, cyano, C1-3Alkyl, monohalo and polyhalo-C1-3Alkyl and C3-6A cycloalkyl group;
R3selected from hydrogen, C1-3Alkyl radical, C3-6Cycloalkyl, monohalo and polyhalo-C1-3Alkyl, homoaryl (homoaryl) and heteroaryl;
X1、X2、X3、X4independently is C (R)4) Or N, provided that no more than two of them represent N; each R4Selected from hydrogen, halo, C1-3Alkyl, monohalo and polyhalo-C1-3Alkyl, cyano, C1-3Alkoxy, monohalo and polyhalo-C1-3An alkoxy group;
l is a bond or-N (R)5) CO-, wherein R5Is hydrogen or C1-3An alkyl group;
ar is a homoaryl or heteroaryl group;
wherein the homoaryl is phenyl or is selected from halo, cyano, C1-3Alkyl radical, C1-3Alkoxy, monohalo and polyhalo-C1-3Phenyl substituted with one, two or three substituents of alkyl;
heteroaryl is selected from pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl and oxadiazolyl, each optionally selected from halo, cyano, C1-3Alkyl radical, C1-3Alkoxy, monohalo and polyhalo-C1-3One, two or three substituents of alkyl; or
Addition salts or solvates thereof.
An illustrative example of the invention is a pharmaceutical composition comprising a pharmaceutically acceptable carrier and any of the compounds described herein. An illustrative example of the invention is a pharmaceutical composition prepared by admixing any of the compounds described herein and a pharmaceutically acceptable carrier. Illustrated herein is a process for the preparation of a pharmaceutical composition comprising admixing any of the compounds described herein and a pharmaceutically acceptable carrier.
Illustrating the invention is a method of treating a disorder mediated by β -secretase, which comprises administering to a subject in need thereof a therapeutically effective amount of any of the compounds or pharmaceutical compositions described herein.
Further illustrated is a method of inhibiting β -secretase of the present invention comprising administering to a subject in need thereof a therapeutically effective amount of any of the compounds or pharmaceutical compositions described herein.
An example of the present invention is a method of treating a disorder selected from the group consisting of alzheimer's disease, mild cognitive impairment, senility, dementia with lewy bodies, cerebral amyloid angiopathy, multi-infarct dementia, down's syndrome, dementia associated with stroke, dementia associated with parkinson's disease and dementia associated with beta amyloid, preferably alzheimer's disease, comprising administering to a subject in need thereof a therapeutically effective amount of any of the compounds or pharmaceutical compositions described herein.
Another example of the invention is any compound described herein for use in treating, in a subject in need thereof: (a) alzheimer's disease, (b) mild cognitive impairment, (c) aging, (d) dementia, (e) dementia with Lewy bodies, (f) Down's syndrome, (g) dementia associated with stroke, (h) dementia associated with Parkinson's disease and (i) dementia associated with amyloid beta.
Detailed Description
The present invention relates to compounds of formula (I) as defined hereinbefore and pharmaceutically acceptable salts and solvates thereof. The compounds of formula (I) are inhibitors of beta-secretase (also known as beta-site cleaving enzyme, BACE1, Asp2 or memapsin2) and are useful in the treatment of alzheimer's disease, mild cognitive impairment, senility, dementia associated with stroke, dementia with lewy bodies, down's syndrome, dementia associated with parkinson's disease and dementia associated with beta amyloid, preferably alzheimer's disease, mild cognitive impairment or dementia, more preferably alzheimer's disease.
In one embodiment of the invention, R1And R2Independently selected from hydrogen, halo, cyano, C1-3Alkyl, monohalo and polyhalo-C1-3Alkyl and C3-6A cycloalkyl group;
R3selected from hydrogen, C1-3Alkyl radical, C3-6Cycloalkyl, monohalo and polyhalo-C1-3Alkyl, homoaryl and heteroaryl;
X1、X2、X3、X4independently is C (R)4) Or N, provided that no more than two of them represent N; each R4Selected from hydrogen, halo, C1-3Alkyl, monohalo and polyhalo-C1-3Alkyl, cyano, C1-3Alkoxy, monohalo and polyhalo-C1-3An alkoxy group;
l is a bond or-N (R)5) CO-, wherein R5Is hydrogen or C1-3An alkyl group;
ar is a homoaryl or heteroaryl group;
wherein the homoaryl is phenyl or is selected from halo, cyano, C1-3Alkyl radical, C1-3Alkoxy, monohalo and polyhalo-C1-3Phenyl substituted with one, two or three substituents of alkyl;
heteroaryl is selected from pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, thiazolyl, thiadiazolyl, oxazolyl and oxadiazolyl, each optionally selected from halo, cyano, C1-3Alkyl radical, C1-3Alkoxy, monohalo and polyhalo-C1-3One, two or three substituents of alkyl; or
Addition salts or solvates thereof.
In one embodiment of the invention, R1And R2Independently selected from hydrogen and C1-3An alkyl group;
R3is C1-3An alkyl group;
X1、X2、X3、X4independently is C (R)4) Wherein each R4Selected from hydrogen and halo;
l is a bond or-N (R)5) CO-, wherein R5Is hydrogen;
ar is a homoaryl or heteroaryl group;
wherein the homoaryl is phenyl or is selected from halo, cyano, C1-3Alkyl and C1-3Phenyl substituted with one or two substituents of alkoxy;
heteroaryl is selected from the group consisting of pyridyl, pyrimidinyl, pyridazinyl and pyrazinyl, each optionally selected from the group consisting of halo, cyano, C1-3Alkyl and C1-3One or two substituents of alkoxy; or
Addition salts or solvates thereof.
In another embodiment of the invention, R1And R2Is hydrogen; r3Is methyl;
X1、X2、X3、X4is CH or CF;
l is a bond or-N (R)5) CO-, wherein R5Is hydrogen;
ar is a homoaryl or heteroaryl group;
wherein the heteroaryl is phenyl substituted with chloro;
heteroaryl is selected from pyridyl, pyrazinyl, pyridazinyl and pyrimidinyl, each optionally substituted with one or two substituents selected from chloro, fluoro, cyano, methyl and methoxy; or
Addition salts or solvates thereof.
In another embodiment, R1And R2Independently selected from hydrogen, mono-, di-and trifluoromethyl, chloro, bromo and cyano;
R3is C1-3Alkyl or mono-, di-and trifluoromethyl;
X1and X3Independently CH or CF; x2And X4Is CH;
l is-N (R)5) CO-, wherein R5Is hydrogen
Ar is a homoaryl or heteroaryl group;
wherein the homoaryl is selected from halo, cyano, C1-3Alkyl and C1-3Phenyl substituted with one or two substituents of alkoxy;
heteroaryl is selected from the group consisting of pyridyl, pyrimidinyl, pyridazinyl, pyrazolyl, oxazolyl and isothiazolyl, each optionally selected from the group consisting of halo, cyano, C1-3Alkyl radical, C1-3Alkoxy, mono-, di-, and trifluoromethyl substituted with one or two substituents; or
Addition salts or solvates thereof.
In another embodiment, R1Is hydrogen, mono-, di-or trifluoromethyl, chloro, bromo or cyano;
R2is hydrogen, chloro, cyano, mono-, di-or trifluoromethyl;
R3is methyl, mono-, di-or trifluoromethyl;
X1is CF; x2、X3、X4Is CH;
l is-N (R)5) CO-, wherein R5Is hydrogen;
ar is heteroaryl;
wherein heteroaryl is selected from pyridyl, pyrazinyl and pyrazolyl, each substituted with one or two substituents selected from chloro, fluoro, cyano, methyl, methoxy, ethoxy, mono-, di-and trifluoromethyl; or
Addition salts or solvates thereof.
In another embodiment, R1And R2Independently selected from hydrogen, mono-, di-and trifluoromethyl, chloro, bromo and cyano;
R3is C1-3Alkyl or mono-, di-and trifluoromethyl;
X1and X3Independently CH or CF; x2And X4Is CH;
l is-N (R)5) CO-, wherein R5Is hydrogen;
ar is heteroaryl;
wherein heteroaryl is selected from the group consisting of 5-chloro-2-pyridyl, 5-fluoro-2-pyridyl, 5-cyano-2-pyridyl, 3, 5-dichloro-2-pyridyl, 3-fluoro-5-chloro-2-pyridyl, 3-fluoro-5-cyano-2-pyridyl, 3-chloro-5-cyano-2-pyridyl, 5-methoxy-2-pyrazinyl, 5-ethoxy-2-pyrazinyl, 1-difluoromethyl-3-pyrazolyl, 2-methyl-4-oxazolyl, 2, 5-dimethyl-4-oxazolyl, 2-methyl-5-trifluoromethyl-4-oxazolyl, and mixtures thereof, 3-isothiazolyl, or
Addition salts or solvates thereof.
In another embodiment, R is3The substituted carbon atom has the R configuration.
Definition of
"halo" refers to fluoro, chloro, and bromo; "C1-3Alkyl "refers to a straight chain having 1,2 or 3 carbon atomsOr branched alkyl groups such as methyl, ethyl, 1-propyl and 2-propyl; "C1-3Alkoxy "shall mean where C is1-3Alkyl groups are ether groups as defined previously; "Mono-and polyhalo-C1-3Alkyl "shall mean C as previously defined substituted by 1,2,3 or, where possible, by more halo atoms as previously defined1-3An alkyl group; "Mono-and polyhalo-C1-3Alkoxy "shall mean an ether group, in which monohalogenated and polyhalogenated C1-3Alkyl is as previously defined; "C3-6Cycloalkyl "refers to cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl;
“C3-6cycloalkanediyl "shall mean divalent radicals such as cyclopropanediyl, cyclobutanediyl, cyclopentanediyl and cyclohexanediyl.
The term "subject" as used herein refers to an animal, preferably a mammal, most preferably a human, who is or has been the subject of treatment, observation or experiment.
The term "therapeutically effective amount" as used herein, means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes alleviation of the symptoms of the disease or disorder being treated.
The term "composition" as used herein is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.
It will be appreciated that certain compounds according to formula (I) and addition salts, hydrates and solvates thereof may contain one or more chiral centers and exist as stereoisomeric forms.
In this context, the term "compound of formula (I)" is intended to include addition salts, solvates and stereoisomers thereof.
The terms "stereoisomer" or "stereochemically isomeric form" are used interchangeably in the present context.
The present invention includes all stereoisomers of the compounds of formula (I) either as pure stereoisomers or as mixtures of two or more stereoisomers.
Enantiomers are mirror image stereoisomers that do not overlap with each other. The 1:1 mixture of a pair of enantiomers is a racemate or a racemic mixture. Diastereomers (diastereomers) are stereoisomers of diastereomers, i.e., they are not mirror images. If the compound contains a double bond, the substituents may be in the E or Z configuration. If the compound contains a disubstituted cycloalkyl group, the substituents may be in the cis or trans configuration. Thus, the present invention includes enantiomers, diastereomers, racemates, E isomers, Z isomers, cis isomers, trans isomers and mixtures thereof.
Absolute configuration was assigned according to the Cahn-Ingold-Prelog system. The configuration on the asymmetric atom is designated as R or S. Resolved compounds with unknown absolute configuration can be designated (+) or (-), depending on the direction in which they rotate plane polarized light.
When a specific stereoisomer is identified, this means that said stereoisomer is substantially free, i.e. involves less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, in particular less than 2% and most preferably less than 1% of other isomers. Thus, for example, when a compound of formula (I) is designated as (R), this means that the compound is substantially free of the (S) isomer; for example, when a compound of formula (I) is designated as E, this means that the compound is substantially free of the Z isomer; for example, when a compound of formula (I) is designated as cis, this means that the compound is substantially free of the trans isomer.
The compounds of formula (I) coexist in dynamic equilibrium with tautomers of formula (I-a).
Moreover, certain crystal forms of the compounds of the present invention may exist as polymorphs and as such are intended to be included in the present invention. In addition, certain compounds of the present invention may form solvates with water (i.e., hydrates) or common organic solvents, and such solvates are also intended to be encompassed within the scope of the present invention.
For pharmaceutical use, salts of the compounds of the present invention refer to non-toxic "pharmaceutically acceptable salts". However, other salts may also be useful in the preparation of the compounds according to the invention or pharmaceutically acceptable salts thereof. Suitable pharmaceutically acceptable salts of the compounds include acid addition salts, which may be formed, for example, by mixing a solution of the compound with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid, or phosphoric acid. Furthermore, where the compounds of the invention bear an acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts, such as sodium or potassium salts; alkaline earth metal salts, for example, calcium or magnesium salts; and salts with suitable organic ligands, for example, quaternary ammonium salts.
Representative acids useful in the preparation of pharmaceutically acceptable salts include, but are not limited to, the following: acetic acid, 2-dichloroacetic acid, acylated amino acids, adipic acid, alginic acid, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, (+) -camphoric acid, camphorsulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, ethane-1, 2-disulfonic acid, ethanesulfonic acid, 2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid, mucic acid, gentisic acid, glucoheptonic acid, D-gluconic acid, D-glucuronic acid, L-glutamic acid, beta-oxo-glutaric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, (+) -L-lactic acid, (±) -DL-lactic acid, lactobionic acid, maleic acid, (-) -L-malic acid, malonic acid, L-malic acid, maleic acid, succinic acid, fumaric acid, maleic, (±) -DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1, 5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+) -L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoromethanesulfonic acid and undecylenic acid. Representative bases that can be used to prepare pharmaceutically acceptable salts include, but are not limited to, the following: ammonia, L-arginine, benzphetamine (benethamine), benzathine (benzathine), calcium hydroxide, choline, dimethylethanolamine, diethanolamine, diethylamine, 2- (diethylamino) -ethanol, ethanolamine, ethylenediamine, N-methyl-glucamine, hydrabamine, 1H-imidazole, L-lysine, magnesium hydroxide, 4- (2-hydroxyethyl) -morpholine, piperazine, potassium hydroxide, 1- (2-hydroxyethyl) -pyrrolidine, secondary amines, sodium hydroxide, triethanolamine, tromethamine and zinc hydroxide.
The names of the compounds of the present invention were generated using the software of Advanced Chemical Development, inc. (ACD/naming products version 10.01; Build 15494, 2006, 12, 1) according to the systematic naming convention agreed upon by the Chemical abstracts Community (CAS) or using the software of Advanced Chemical Development, inc. (ACD/naming products version 10.01.0.14105, 2006, 10) according to the systematic naming convention agreed upon by the international union of theory and applied chemistry (IUPAC). In the case of tautomeric forms, the name of the tautomeric form of the depicted structure is generated. Other tautomeric forms not described are also included within the scope of the invention.
A. Preparation of the Final Compounds
Experimental procedures 1
For example, the final compound according to formula (I) may be prepared according to reaction scheme (1) by reacting the intermediate compound of formula (II) with a suitable source of ammonia such as ammonium chloride or aqueous ammonia, the reaction being carried out under thermal conditions such as, for example, heating the reaction mixture at 60-90 ℃ for, for example, 6-100 hours, in a suitable reaction inert solvent such as, for example, water or methanol. In reaction scheme (1), all variables are as defined in formula (I).
Reaction scheme 1
Experimental procedures 2
Intermediates of formula (IV) wherein L is-N (R) can be prepared according to reaction scheme (2) by reacting an intermediate compound of formula (III-a) with an intermediate of formula (IV)5) The final compound according to formula (I-a) of CO-, is reacted in a suitable base such as, for example, K3PO4Copper catalysts such as, for example, CuI and diamines such as, for example, (1R,2R) - (-) -1, 2-diaminocyclohexane, in the presence of a catalyst such as, for example, copper, in a suitable reaction-inert solvent such as, for example, N-dimethyl-formamide, are heated under thermal conditions such as, for example, microwave irradiation at 180 ℃ for, for example, 135 minutes. In reaction scheme (2) all variables are as defined for formula (I) and W is halo.
Reaction scheme 2
Experimental procedures 3
Furthermore, the final compound according to formula (I-a) may be prepared according to reaction scheme (3) by reacting an intermediate compound of formula (III-b) with an intermediate of formula (V), the reaction being stirred under thermal conditions, e.g. at 25 ℃, for e.g. 2-18 hours, optionally in the presence of a suitable base such as, e.g. N, N-diisopropylethylamine and in the presence of a condensing agent such as, e.g. 2- (1H-7-aza-benzotriazol-1-yl) -N, N' -tetramethyluronium hexafluorophosphate [ HATU, CAS 148893-10-1] or 4- (4, 6-dimethoxy-1, 3, 5-triazin-2-yl) -4-methylmorpholinium chloride [ DMTMM, CAS 3945-69-5] in a suitable reaction-inert solvent, such as, for example, dichloromethane or methanol. In reaction scheme (3), all variables are as defined in formula (I).
Reaction scheme 3
Experimental procedures 4
Furthermore, the final compound according to formula (I-a) may be prepared according to reaction scheme (4) by reacting an intermediate compound of formula (III-b) with an intermediate of formula (VI), the reaction being carried out under thermal conditions such as, for example, 25 ℃ with stirring of the reaction mixture, for example for 2 hours, in the presence of a suitable base such as, for example, pyridine, in a suitable reaction-inert solvent such as, for example, dichloromethane. In reaction scheme (4), all variables are as defined in formula (I) and Y is halo.
Reaction scheme 4
Experimental procedures 5
The final compound according to formula (I-b) wherein L is a bond may be prepared according to reaction scheme (5) by reacting an intermediate compound of formula (III-a) with an intermediate of formula (VII), the reaction being under thermal conditions such as, for example, under microwave irradiation, heating the reaction mixture at 80 ℃ for, for example, 20 hours or, for example, 150 ℃ for 10 minutes to 30 minutes, in a suitable base such as, for example, K2CO3Pd-complex catalysts such as, for example, tetrakis (triphenylphosphine) palladium (0) [ CAS 14221-01-3]In the presence of a suitable reaction inert solvent such as, for example, a mixture of inert solvents such as, for example, 1, 4-dioxane/ethanol. In reaction scheme (5), all variables are as defined in formula (I) and W is halo. R6And R7May be hydrogen or alkyl, or may together form, for example, the formula-CH2CH2-、-CH2CH2CH2-or-C (CH)3)2C(CH3)2A divalent group of (a).
Reaction scheme 5
Various intermediates and starting materials in the foregoing preparations are known compounds which can be prepared according to methods known in the art for the preparation of the or similar compounds, and certain intermediates are novel. A number of such methods of preparation are described in more detail below.
Experimental procedures 6
Alternatively, compounds of formula (I-c) wherein Y is Br or I can be reacted with zinc cyanide and sodium cyanide according to reaction scheme (6) to produce compounds wherein R is1The final compound according to formula (I-a), which is CN, referred to herein as a compound of formula (I-d), is reacted by heating the reaction mixture under thermal conditions such as, for example, 110 ℃ for, for example, 16 to 21 hours in the presence of a suitable coupling agent such as, for example, tetrakis (triphenylphosphine) palladium (0), in a mixture of a suitable reaction inert solvent such as, for example, dimethylformamide and toluene. In reaction scheme (6) all variables are as defined in formula (I) and Z1Is a protecting group for an amine such as, for example, tert-butoxycarbonyl.
Reaction scheme 6
Experimental procedures 7
Furthermore, the final compound according to formula (I-a) may be prepared by deprotecting the intermediate compound of formula (I-e) according to reaction scheme (7) with a suitable acid such as, for example, trifluoroacetic acid, and the reaction is carried out under thermal conditions such as, for example, 25 ℃ with stirring of the reaction mixture, for example, for 30 minutes, in a suitable reaction inert solvent such as, for example, dichloromethane. In reaction scheme (7) all variables are as defined in formula (I) and Z1Is a protecting group for an amine such as, for example, tert-butoxycarbonylAnd (4) a base.
Reaction scheme 7
B. Preparation of intermediate compounds
Experimental procedures 8
Intermediates according to formula (II) can be prepared according to reaction scheme (8) by reacting an intermediate compound of formula (VIII) with a sulfur donating agent suitable for thioamide synthesis such as, for example, phosphorus pentasulfide or 2, 4-bis- (4-methoxyphenyl) -1, 3-dithia-2, 4-diphosphetane 2, 4-disulfide [ Lawesson's reagent, CAS 19172-47-5], under thermal conditions such as, for example, 90 ℃ heating the reaction mixture, for example, for 18 hours, in a reaction inert solvent such as, for example, tetrahydrofuran or toluene, optionally in the presence of a suitable base such as, for example, pyridine. In reaction scheme (6), all variables are as defined in formula (I).
Reaction scheme 8
Experimental procedures 9
Intermediates according to formula (VIII) wherein L is a bond, herein referred to as intermediates of formula (VIII-a), may be prepared according to reaction scheme (9) by reacting an intermediate compound of formula (IX-a) with an intermediate of formula (VII), under thermal conditions, e.g. under microwave irradiation, heating the reaction mixture at 80 deg.C, e.g. for 20 hours or e.g. 150 deg.C, e.g. for 15-30 minutes, in the presence of a suitable base, e.g. aqueous Na2CO3Pd-complex catalysts such as, for example, tetrakis (triphenylphosphine) palladium (0) [ CAS 14221-01-3]In the presence of a suitable inert solvent, e.g. 1, 4-bisOxacyclohexane/water. In reaction scheme (7), all variables are as defined in formula (I) and W is halo. R6And R7May be hydrogen or alkyl, or may together form, for example, the formula-CH2CH2-、-CH2CH2CH2-or-C (CH)3)2C(CH3)2A divalent group of (a).
Reaction scheme 9
Experimental procedures 10
Intermediates according to formula (III-b) can be prepared from the corresponding intermediate compounds of formula (III-a) according to reaction scheme (10) following art-known coupling procedures of the Buchwald-Hartwig type. Said coupling being effected under thermal conditions such as, for example, heating the reaction mixture under microwave irradiation, for example at 80 ℃ for, for example, 20 hours or, for example, at 130 ℃ for, for example, 10 minutes, in a suitable base such as, for example, aqueous K3PO4Or Cs2CO3Pd-complex catalysts such as, for example, [1, 1' -bis (diphenylphosphino) ferrocene]Dichloropalladium (II) [ CAS 72287-26-4]Or trans-bis (dicyclohexylamine) palladium diacetate [ DAPCy, CAS 628339-96-8]By treating the intermediate compound of formula (III-a) with an intermediate of formula (X-a) in the presence of a suitable reaction inert solvent such as, for example, ethanol or a mixture of inert solvents such as, for example, 1, 2-dimethoxy-ethane/water/ethanol. In reaction scheme (8), all variables are as defined in formula (I) and W is halo. R5Is hydrogen or C1-3An alkyl group.
Reaction scheme 10
Experimental procedureSequence of steps 11
Alternatively, R can be prepared according to reaction scheme (11) from the corresponding intermediate of formula (III-c) following art-known nitro to amino reduction procedures5Is an intermediate according to formula (III-b) of hydrogen. The reduction may be conveniently carried out according to catalytic hydrogenation procedures known in the art. For example, the reduction may be carried out by stirring the reactants under a hydrogen atmosphere and in the presence of a suitable catalyst such as, for example, palladium on carbon, platinum on carbon, raney nickel, etc. Suitable solvents are, for example, water, alkanols, such as methanol, ethanol, etc., esters, such as ethyl acetate, etc. To increase the speed of the reduction reaction, the temperature and/or pressure of the reaction mixture may advantageously be increased. Further hydrogenation of certain functional groups in the reactants and reaction products that are undesirable can be prevented by adding catalyst poisons such as, for example, thiophenes and the like to the reaction mixture. In reaction scheme (11), all variables are as defined in formula (I).
Reaction scheme 11
Experimental procedures 12
Intermediates according to formula (III-b) can be prepared from the corresponding intermediates of formula (III-a) according to reaction scheme (12) following art-known Buchwald-Hartwig type coupling procedures between intermediates of formula (III-a) and (X-b) to give intermediates of formula (III-d), followed by hydrolysis of (III-d) to give (III-a). The Buchwald-Hartwig coupling can be carried out by heating the reaction mixture under thermal conditions such as, for example, 90 ℃ for, for example, 18 hours in the presence of a suitable base such as, for example, sodium tert-butoxide, a Pd-complex catalyst such as, for example, tris (dibenzylideneacetone) dipalladium (0) [ Pd ]2(dba)3,CAS 51364-51-3]Phosphine ligands such as, for example, rac-2,2 '-bis (diphenylphosphino) -1,1' -binaphthyl [ rac-BINAP, CAS98327-87-8]In the presence of a suitable reaction inert solventSuch as, for example, toluene, by treating the intermediate compound of formula (III-a) with an intermediate of formula (X-b). (III-d) may be hydrolyzed to (III-a) by treatment with HCl in 2-propanol under acidic conditions, e.g., at room temperature for 1-4 hours. In reaction scheme (12), all variables are as defined in formula (I) and W is halo. R5Is a diphenylmethylene group.
Reaction scheme 12
Experimental procedures 13
Intermediates according to formula (III-b) can be prepared from the corresponding intermediate compounds of formula (III-a) according to reaction scheme (13) by heating the reaction mixture under thermal conditions such as, for example, 110 deg.C for, for example, 3-6 hours, under sodium azide [ CAS 26628-22-8]Suitable copper salts are, for example, copper (I) iodide [ CAS 7681-65-4 ]]Suitable bases are, for example, Na2CO3And suitable diamine ligands such as, for example, N, N' -dimethylethylenediamine [ CAS 110-70-3]In the presence of a suitable reaction-inert solvent, such as, for example, dimethyl sulfoxide. In reaction scheme (13), all variables are as defined in formula (I) and W is halo.
Reaction scheme 13
Experimental procedures 14
The intermediate compounds of formula (III-a) and the intermediate compounds of formula (III-c) can be prepared generally according to the reaction procedures shown in the following reaction schemes (14) and (15).
Reaction scheme 14
A: conversion of thioamides to amidines
B: conversion of amides to thioamides (sulfurization)
C and H: cyclization of
D: removal of any N-protecting group Z1
E: alkylation
The amidine derivatives of scheme (14) above may be conveniently prepared from the corresponding thioamide derivatives following art-known procedures for conversion of thioamides to amidines (reaction step a). The conversion may conveniently be carried out by treating the thioamide with an ammonia source such as, for example, ammonium chloride or aqueous ammonia under thermal conditions such as, for example, heating the reaction mixture at 60-90 c for, for example, 6-100 hours in a suitable reaction inert solvent such as, for example, water or methanol and the like.
Alternatively, the amidine derivatives of scheme (14) above can be prepared from the corresponding intermediate compounds of formula (XIII-d) following art-known cyclization procedures (reaction step H). The cyclisation may conveniently be carried out by treating the intermediate compound of formula (XIII-d) with a suitable acid, such as 4M hydrochloric acid in dioxane or TFA, under thermal conditions such as, for example, heating the reaction mixture between 25 ℃ and 70 ℃ for, for example, 2 to 5 hours.
The thioamide derivatives of reaction scheme (14) above can be prepared from the amide derivatives following art-known sulfurization procedures (reaction step B). The conversion may conveniently be carried out by treating the amide with a sulfurizing agent such as, for example, phosphorus pentasulfide or 2, 4-bis- (4-methoxy-phenyl) -1, 3-dithia-2, 4-diphosphetane 2, 4-disulfide [ Lawesson's reagent, CAS 19172-47-5] in the presence of a suitable base such as pyridine, in a reaction inert solvent such as, for example, tetrahydrofuran or 1, 4-dioxane, or the like, under thermal conditions such as, for example, 50-100 ℃ for, for example, 24 hours.
The amide derivatives of formula (IX-a) and (IX-C) in reaction scheme (14) above can be prepared from the corresponding intermediate compounds of formula (XII-a) and (XII-C) following art-known cyclization procedures (reaction step C). The cyclisation may conveniently be carried out by treating the intermediate compounds of formulae (XII-a) and (XII-c) with a suitable base, such as sodium methoxide or potassium carbonate, in a suitable reaction solvent such as, for example, methanol and the like, at-80 ℃ to 100 ℃, preferably-15 ℃ to 60 ℃, for 30 minutes to 100 hours, preferably 1 to 24 hours. Alternatively, standard conditions for amide formation from esters can be employed by treating intermediate compounds of formula (XII-a) and (XII-c) with a Lewis acid, such as, for example, trimethylaluminum, under thermal conditions, such as, for example, heating the reaction for 30 minutes at 120 ℃ with microwave irradiation, in a suitable inert solvent, such as, for example, tetrahydrofuran.
May be prepared by removing the protecting group Z according to procedures known in the art1The intermediate compounds of the formulae (XII-a) and (XII-c) in the above reaction scheme (14) are prepared from the corresponding intermediate compounds of the formulae (XIII-a) and (XIII-c).
Reaction scheme 15
E: alkylation
F: oxathiazole alkoxylation
G: oxathiazolidine formation
The alkylation procedure known in the art (reaction step E) can be followed from Z therein1Are amine protecting groups such as, for example, tert-butoxycarbonyl, the corresponding intermediate compounds of formulae (XV-a) and (XV-c) preparing intermediates according to formulae (XIII-a), (XIII-c) and (XIII-d) in reaction schemes (14) and (15) above. The alkylation may be carried out at low temperatures such as, for example, 0 ℃ for 30 minutes and then at temperatures such as, for exampleFrom 24 hours to 100 hours at a temperature of from 60 ℃ to 100 ℃ or, for example, heating the reaction mixture, for example from 30 minutes to 45 minutes, in a suitable inert solvent such as, for example, N, N-dimethylformamide, acetonitrile or tetrahydrofuran, by reaction with a suitable base such as, for example, sodium hydride, cesium carbonate, potassium carbonate or 1, 8-diazabicyclo [5.4.0 ]]The treatment of XIV or XVIII with the corresponding intermediate compounds of formulae (XV-a) and (XV-c) is conveniently carried out with undec-7-ene.
Intermediates according to formulae (XV-a) and (XV-c) in reaction scheme (15) above may be prepared by reacting intermediate compounds of formulae (XVI-a) and (XVI-c) according to oxidation procedures known in the art (reaction step F). The catalyst may be used in the presence of a catalyst such as, for example, ruthenium (III) chloride [ CAS: 10049-08-8] in the presence of a suitable inert solvent such as, for example, acetonitrile/water, the oxidation is conveniently carried out by treating the corresponding intermediate compounds of formulae (XVI-a) and (XVI-c) with an oxidizing agent such as, for example, sodium periodate.
Intermediates according to formulae (XVI-a) and (XVI-c) in reaction scheme (15) above can be prepared by reacting intermediate compounds of formulae (XVII-a) and (XVII-c) according to sulfonamide lactone (sulfonamide) formation procedures known in the art (reaction step G). The conversion may conveniently be carried out by treating the corresponding intermediate compounds of formula (XVII-a) and (XVII-c) with thionyl chloride at low temperatures such as, for example, -40 ℃ for example 30 minutes and then at temperatures such as, for example, 25 ℃ for example 24 to 72 hours in a suitable reaction inert solvent such as, for example, acetonitrile in the presence of a base such as, for example, pyridine.
In general, the preparation of Z therein can be carried out according to the Strecker-type procedure described in the literature known in the art1Are amine protecting groups such as, for example, tert-butoxycarbonyl intermediate compounds of formulas (XVII-a) and (XVII-c).
Experimental procedures 16
The intermediate compound of formula (XVIII) can be generally prepared according to the reaction procedure represented in the following reaction scheme (16).
Reaction scheme 16
Can be prepared by reacting an intermediate compound of formula (XIX) wherein Z is2The cyano derivative of formula (XVIII) in reaction scheme (16) above is conveniently prepared by deprotection of a protecting group that is imidazole such as, for example, 2- (trimethylsilyl) ethoxymethyl). The deprotection can be conveniently carried out by treatment with tetrabutylammonium fluoride under thermal conditions such as, for example, heating the reaction mixture at 65 ℃ for, for example, 4 hours.
The intermediate according to formula (XIX) in reaction scheme (16) above may be prepared by reacting the intermediate compound of formula (XX) with a suitable acid such as, for example, acetic anhydride, under thermal conditions such as, for example, heating the reaction mixture at 140 ℃ for, for example, 6 hours.
The intermediate according to formula (XX) in reaction scheme (16) above may be prepared by reacting the intermediate compound of formula (XXI) with hydroxylamine hydrochloride in the presence of a suitable reaction inert solvent such as, for example, distilled water, under thermal conditions such as, for example, heating the reaction mixture at 70 ℃ for, for example, 1 hour.
Intermediates according to formula (XXI) in reaction scheme (16) above can be prepared by reacting intermediate compounds of formula (XXII) according to carbonylation procedures known in the art. The carbonylation may be conveniently carried out by treating the corresponding intermediate of formula (XXII) with dimethylformamide under thermal conditions such as, for example, -78 ℃ cooling the reaction mixture, for example 1-2 hours, in the presence of a base such as, for example, butyllithium or lithium diisopropylamide, in a suitable reaction-inert solvent such as, for example, tetrahydrofuran.
The intermediate according to formula (XXII) in reaction scheme (16) above may be prepared by protecting an intermediate compound of formula (XXIII). The protection can be conveniently carried out by treating the corresponding intermediate of formula (XXIII) with 2- (trimethylsilyl) ethoxymethyl chloride under thermal conditions such as, for example, stirring the reaction mixture at 25 ℃ for, for example, 30 minutes in a suitable reaction-inert solvent such as, for example, tetrahydrofuran.
Intermediates according to formula (XIII) in reaction scheme (16) above are commercially available.
Experimental procedures 17
Wherein R can be prepared in general by the reaction procedure shown in the following reaction scheme (17)1AIs H or Cl and R2Intermediate compounds of formula (III-b) which are CN, are referred to herein as intermediates of formula (III-e).
Reaction scheme 17
The intermediate compounds of formula (XXIV) (wherein Z is1Is a protecting group of an amine such as, for example, t-butoxycarbonyl), an intermediate according to formula (III-e) in reaction scheme (17) above is conveniently prepared. The deprotection may conveniently be carried out by stirring the reaction mixture under thermal conditions such as, for example, 25 ℃ for, for example, 30 minutes, by treatment with trifluoroacetic acid in the presence of a suitable reaction-inert solvent such as, for example, dichloromethane.
Intermediate compounds according to formula (XXIV) can be prepared according to reaction scheme (17) by reacting an intermediate compound of formula (XXV) with zinc cyanide and zinc, which reaction can be carried out under thermal conditions such as, for example, heating the reaction mixture under microwave irradiation at 150 ℃ for, for example, 30 minutes, in a suitable reaction-inert solvent such as, for example, dimethylacetamide, in the presence of a suitable ligand such as, for example, 1,1' -bis (diphenylphosphino) ferrocene, in the presence of a suitable coupling agent such as, for example, tris (dibenzylideneacetone) dipalladium (0).
Intermediates according to formula (XXV) in reaction scheme (17) above can be conveniently prepared by iodinating intermediate compounds of formula (III-f) according to procedures known in the art. The halogenation may be carried out under thermal conditions such as, for example, -78 ℃ cooling of the reaction mixture, for example, for 10 minutes, in a reaction-inert solvent such as, for example, tetrahydrofuran, in a suitable base such as, for exampleNConveniently by treatment with iodine in the presence of butyllithium.
Intermediates according to formula (III-f) in reaction scheme (17) above can be prepared by protecting intermediate compounds of formula (XXVI). The protection can be conveniently carried out by treating the corresponding intermediate of formula (XXVI) with di-tert-butyl dicarbonate under thermal conditions such as, for example, stirring the reaction mixture at 25 ℃ for, for example, 24 hours in a suitable reaction-inert solvent such as, for example, dichloromethane in the presence of a base such as, for example, N-diisopropylethylamine.
In reaction scheme (17), R1AIs H, Cl and all other variables are as defined in formula (I).
Experimental procedures 18
Wherein R can be prepared in general by the reaction procedure shown in the following reaction scheme (18)1Is H, R2Is CF2Is referred to herein as an intermediate of formula (III-g).
Reaction scheme 18
The intermediate compounds of formula (XXVII) wherein Z is as defined in the description may be prepared by procedures known in the art1Is a protecting group of an amine such as, for example, t-butoxycarbonyl), an intermediate according to formula (III-g) in reaction scheme (18) above is conveniently prepared. The de-protectionThe reaction mixture is stirred under thermal conditions, for example at 25 ℃ for, for example, 30 minutes, and treatment with trifluoroacetic acid in the presence of a suitable reaction-inert solvent such as, for example, dichloromethane, is conveniently carried out.
The intermediate compound according to formula (XXVII) may be prepared according to reaction scheme (18) by reacting the intermediate compound of formula (XXVIII) with diethylaminosulfur trifluoride in a suitable reaction-inert solvent such as, for example, dichloromethane, under thermal conditions such as, for example, stirring the reaction mixture at 25 ℃ for, for example, 16 hours.
Intermediates according to formula (XXVIII) in reaction scheme (18) above can be conveniently prepared by oxidation of intermediate compounds of formula (XXIX) according to procedures known in the art. The oxidation is conveniently carried out by heating the reaction mixture under thermal conditions such as, for example, 25 ℃ for, for example, 2 hours, in a reaction inert solvent such as, for example, dichloromethane, treated with manganese dioxide.
Intermediates according to formula (XXIX) in reaction scheme (18) above may conveniently be prepared by hydroxymethylation of intermediate compounds of formula (XXX) according to procedures known in the art. The methylolation may be carried out by cooling the reaction mixture under thermal conditions such as, for example, -78 ℃ for, for example, 10 minutes in a suitable base such as, for exampleNButyl lithium is conveniently carried out by treatment with paraformaldehyde in an inert solvent such as, for example, tetrahydrofuran.
Intermediates according to formula (XXX) in reaction scheme (18) above may be prepared by protecting intermediate compounds of formula (III-h). Said protection can be conveniently carried out by treating the corresponding intermediate of formula (III-h) with di-tert-butyl carbonate in the presence of a base such as, for example, N-diisopropylethylamine in a suitable reaction inert solvent such as, for example, dichloromethane under thermal conditions such as, for example, stirring the reaction mixture at 25 ℃ for, for example, 24 hours.
In reaction scheme (18), Z1Are protecting groups for amines such as, for example, tert-butoxycarbonyl and all other variantsThe amount is as defined in formula (I).
Experimental procedures 19
The intermediate compounds of formula (I-e) can be prepared generally according to the reaction procedure shown in the following reaction scheme (19).
Reaction scheme 19
Intermediates according to formula (XXXI) in reaction scheme (19) above may be prepared by protection of intermediate compounds of formula (III-b). The protection can be conveniently carried out by treating the corresponding intermediate of formula (III-b) with di-tert-butyl dicarbonate under thermal conditions such as, for example, stirring the reaction mixture at 25 deg.C, for example, for 24 hours, in a suitable reaction-inert solvent such as, for example, dichloromethane.
Intermediates according to formula (I-e) in reaction scheme (19) above may be prepared according to reaction scheme (19) by reacting an intermediate compound of formula (XXXI) with an intermediate of formula (V), which reaction may be carried out under thermal conditions such as, for example, 25 ℃ with stirring of the reaction mixture, for example, for 2 to 18 hours, in a suitable reaction-inert solvent such as, for example, dichloromethane or methanol, in the presence of a suitable base such as, for example, N-diisopropylethylamine, in the presence of a condensing agent such as, for example, 2-, (ii)1H-7-azabenzotriazol-1-yl) -N,N,N ’ ,N ’Tetramethyluronium hexafluorophosphate [ HATU, CAS 148893-10-1]Or 4- (4, 6-dimethoxy-1, 3, 5-triazin-2-yl) -4-methylmorpholinium chloride [ DMTMM, CAS 3945-69-5]In the presence of (a). In reaction scheme (19), Z1Are suitable N-protecting groups and all other variables are as defined in formula (I).
Experimental procedures 20
Wherein R can be prepared according to the reaction procedure shown in the following reaction scheme (20)1Is Cl, R2Is Cl of formula (III-b)) Intermediate compounds, referred to herein as intermediates of formula (III-j).
Reaction scheme 20
The intermediate according to formula (III-j) in reaction scheme (20) above may be prepared by chlorinating the intermediate compound of formula (III-i). The chlorination can be conveniently carried out by treating the corresponding intermediate of formula (III-i) with N-chlorosuccinimide under thermal conditions such as, for example, heating the reaction mixture at 80 ℃, for example, for 16 hours, in a suitable reaction-inert solvent such as, for example, acetic acid. In reaction scheme (20), all variables are as defined in formula (I).
Pharmacology of
The compounds of the present invention and pharmaceutically acceptable compositions thereof inhibit BACE and are therefore useful in the treatment or prevention of Alzheimer's Disease (AD), Mild Cognitive Impairment (MCI), senility, dementia with lewy bodies, cerebral amyloid angiopathy, multi-infarct dementia, down's syndrome, dementia associated with parkinson's disease and dementia associated with beta-amyloid.
The present invention relates to a compound according to general formula (I), a stereoisomeric form thereof or a pharmaceutically acceptable acid or base addition salt or solvate thereof, for use as a medicament.
The invention also relates to a compound according to general formula (I), a stereoisomeric form thereof or a pharmaceutically acceptable acid or base addition salt or solvate thereof, for use in the treatment or prevention of a disease or disorder selected from the group consisting of AD, MCI, senility, dementia with lewy bodies, cerebral amyloid angiopathy, multi-infarct dementia, down's syndrome, dementia associated with parkinson's disease and dementia associated with beta-amyloid.
The invention also relates to the use of a compound according to general formula (I), its stereoisomeric forms or a pharmaceutically acceptable acid or base addition salt or solvate thereof, for the manufacture of a medicament for the treatment or prevention of any of the disease steps mentioned above.
In view of the efficacy of the compounds of formula (I), there is provided a method of treating warm-blooded animals, including humans, suffering from or a method of preventing warm-blooded animals, including humans, suffering from any of the diseases described above.
Said method comprising the administration, i.e. systemic or local administration, preferably oral administration, of an effective amount of a compound of formula (I), its stereoisomeric forms, its pharmaceutically acceptable addition salts or solvates, to warm-blooded animals, including man.
The method of treatment may also comprise administering the active ingredient on a regimen of 1-4 intakes per day. In these methods of treatment, it is preferred to formulate the compounds according to the invention prior to administration. Suitable pharmaceutical formulations are prepared by known procedures using well known and readily available ingredients, as described hereinafter.
The compounds of the present invention suitable for treating or preventing Alzheimer's disease or symptoms thereof may be administered alone or in combination with one or more other therapeutic agents. Combination therapy includes administration of a single pharmaceutical dosage formulation containing a compound of formula (I) and one or more other therapeutic agents as well as administration of a compound of formula (I) and other therapeutic agents each present in its own separate pharmaceutical dosage formulation. For example, the compound of formula (I) and therapeutic agent may be administered in a single oral dosage composition such as a tablet or capsule, or each drug may be administered in separate oral dosage formulations.
Pharmaceutical composition
The present invention also provides compositions for the prevention or treatment of diseases in which inhibition of beta-secretase is beneficial, such as Alzheimer's Disease (AD), mild cognitive impairment, senility, dementia with Lewy bodies, Down's syndrome, dementia associated with stroke, dementia associated with Parkinson's disease and dementia associated with beta-amyloid. The composition comprises a therapeutically effective amount of a compound according to formula (I) and a pharmaceutically acceptable carrier or diluent.
Although it is possible to administer the active ingredient alone, it is preferably presented as a pharmaceutical composition. Accordingly, the present invention further provides a pharmaceutical composition comprising a compound according to the present invention together with a pharmaceutically acceptable carrier or diluent. A carrier or diluent must be "acceptable" in the sense of being compatible with the other ingredients of the composition and not deleterious to the recipient thereof.
The pharmaceutical compositions of the present invention may be prepared by any method well known in the art of pharmacy. A therapeutically effective amount of a particular compound, in base form or addition salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which may take a wide variety of forms depending on the form of preparation desired for administration. It is contemplated that these pharmaceutical compositions are preferably in a form suitable for systemic administration such as oral, transdermal or parenteral administration; or topically administering a single dosage form such as via inhalation, nasal spray, eye drops or via cream, gel, shampoo, etc. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs and solutions; or in the case of powders, pills, capsules and tablets, solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like. Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will typically comprise sterile water, at least in large part, although other ingredients may be included, for example to aid solubility. For example, injectable solutions may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. In compositions suitable for transdermal administration, the carrier optionally comprises a penetration enhancer and/or a suitable humectant, optionally in combination with suitable additives of any nature, in minor proportions, which additives do not cause any significant deleterious effect on the skin. The additives may facilitate administration to the skin and/or may aid in the preparation of the desired composition. These compositions can be administered in various ways, e.g., as a transdermal patch, as a spot-on, or as an ointment.
It is particularly advantageous to formulate the above-described pharmaceutical compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein the specification and claims refer to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect, in association with a required pharmaceutical carrier. Examples of such dosage unit forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, injectable solutions or suspensions, teaspoonfuls, tablespoonfuls and the like, and segregated multiples thereof.
The exact dosage and frequency of administration will depend upon the particular compound of formula (I) employed, the particular condition being treated, the severity of the disease being treated, the age, weight, sex, extent of disorder and general physical condition of the particular patient and other drugs that the individual may be taking, as is well known to those skilled in the art. Moreover, it will be apparent that the effective daily dose may be reduced or increased as determined by the response of the subject being treated and/or as assessed by the physician prescribing the compounds of the instant invention.
Depending on the mode of administration, the pharmaceutical composition will contain from 0.05 to 99% by weight, preferably from 0.1 to 70% by weight, more preferably from 0.1 to 50% by weight of the active ingredient and from 1 to 99.95% by weight, preferably from 30 to 99.9% by weight, more preferably from 50 to 99.9% by weight of a pharmaceutically acceptable carrier, all percentages being based on the total weight of the composition.
The compounds of the invention may be used for systemic administration such as oral, transdermal or parenteral administration; or topical administration such as via inhalation, nasal spray, eye drops or via cream, gel, shampoo, etc. Oral compounds are preferred. It is well known to those skilled in the art that the exact dose and frequency of administration will depend on the particular compound according to formula (I) used, the particular condition being treated, the severity of the disease being treated, the age, weight, sex, degree of disorder and general physical condition of the particular patient and other drugs that the individual may be taking. Moreover, it will be apparent that the effective daily dose may be reduced or increased depending upon the response of the subject being treated and/or as assessed by the physician prescribing the compounds of the instant invention.
The amount of a compound of formula (I) that can be combined with carrier materials to produce a single dosage form will vary depending upon the disease to be treated, the type of mammal, and the particular mode of administration. However, as a general guide, for example, a suitable unit dose of a compound of the invention may preferably contain from 0.1 mg to about 1000 mg of the active compound. A preferred unit dose is from 1 mg to about 500 mg. More preferred unit doses are from 1 mg to about 300 mg. Even more preferred unit doses are from 1 mg to about 100 mg. Such dosage units may be administered more than once a day, e.g., 2,3, 4, 5 or 6 times daily, but preferably 1 or 2 times daily, so that the total dose for a 70 kg adult is in the range of 0.001 to about 15 mg per kg body weight of the subject per administration. The preferred dosage is 0.01 to about 1.5 mg per kg body weight of the subject per administration, and such treatment may extend over weeks or months, and in some cases years. It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed; the age, weight, general health, sex and daily diet of the individual to be treated; the number and route of administration; the rate of drainage; other drugs that have been previously administered; and the severity of the disease under treatment.
In some instances, it may be desirable to employ dosages outside of these ranges, as will be apparent to those skilled in the art. Further, it should be noted that the clinician or attending physician will know how and when to initiate, interrupt, adjust or terminate therapy in connection with the individual patient's response.
The following examples are intended to illustrate but not limit the scope of the invention.
Experimental part
Hereinafter, the term ' m.p. "means melting point, ' THF ' means tetrahydrofuran, DIPE is diisopropyl ether, ' DMF ' means N, N-dimethylformamide, ' DCM ' means dichloromethane, ' EtOAc ' means ethyl acetate," AcOH "means acetic acid," MeOH "means methanol," EtOH "means ethanol," rac "means racemic. SFC is supercritical fluid chromatography, "PFA" means perfluoroalkoxy. "DIPEA" means N, N-diisopropylethylamine, "DIPE" means diisopropyl ether, "DMA" means dimethylacetamide, and "DAST" means diethylaminosulfur trifluoride.
Thin Layer Chromatography (TLC) was performed on silica gel 60F 254 plates (Merck) using a reagent grade solvent. Automated FLASH column chromatography was performed on irregular silica gel of 5-40 μm particle size using a pre-connected (ready-to-connect) cartridge from Merck (normal phase disposable FLASH column) with either SPOT or FLASH system from Armen Instrument.
In a single mode reactor: emrysTMThe reaction with microwave assistance was carried out in an Optimizer microwave reactor (Personal Chemistry a.b., now Biotage).
The hydrogenation reactions were carried out in continuous gas stream hydrogenators H-CUBE from Thales Nano technology Inc.
The fluid reactions were carried out in a commercially available Vapourtec R2+ R4 modular unit with cold reactor modules. Website: http:// www.vapourtec.co.uk.
The absolute configuration of the chiral center (designated as R and/or S) is determined for key intermediates, as well as for certain final compounds, by comparison with samples of known configuration, or using analytical techniques suitable for determining absolute configuration, such as VCD (vibrational circular dichroism) or X-ray crystallography. When the absolute configuration at the chiral center is unknown, it is arbitrarily designated as R or S.
A.Preparation of intermediates
Examples A1
rac-2- Amino group -2-(3- Bromine compound - Phenyl radical )- Preparation of propionitrile
Trimethylsilyl cyanide (20 g,200 mmol) was added to 3-bromo-acetophenone (20 g, 100 mmol) and NH4Cl (11 g,200 mmol) in NH3In stirred solution in MeOH (400 mL). The mixture was stirred at room temperature for 4 days. The solvent was evaporated in vacuo and the residue taken up in EtOAc (100 mL). The solid was filtered off and the filtrate evaporated in vacuo to yield rac-2-amino-2- (3-bromo-phenyl) -propionitrile (20 g, 86% yield) which was used in the next step without further purification.
Examples A2
rac-2- Amino group -2-(3- Bromine compound - Phenyl radical )- Preparation of methyl propionate
Rac-2-amino-2- (3-bromo-phenyl) -propionitrile (20 g, 88.9 mmol) was dissolved in HCl/MeOH (500 mL) and the mixture refluxed for 4 days. After cooling to room temperature, EtOAc (100 mL) and water (100 mL) were added and the mixture was extracted with EtOAc (2 × 100 mL). The combined aqueous layers were basified with aqueous ammonia solution until pH 8 and extracted with EtOAc (5 × 100 mL). Drying (Na)2SO4) The combined organic layers were filtered and the solvent evaporated in vacuo to yield rac-2-amino-2- (3-bromo-phenyl) -propionic acid methyl ester as an oil (10.6 g, 46% yield).
Examples A3
rac-2- Amino group -2-(3- Bromine compound - Phenyl radical )- C3 -1- Preparation of alcohols
Lithium aluminum hydride (1M in THF; 22 mL, 22 mmol) was added dropwise to a stirred solution of rac-2-amino-2- (3-bromo-phenyl) -propionic acid methyl ester (7.5 g, 29.1 mmol) in THF (200 mL) at-15 ℃. The mixture was warmed to 0 ℃ over 1 hour. More THF (150 ml) was added and saturated Na was added dropwise2SO4Until no more hydrogen is formed. Adding anhydrous Na2SO4And stirred at room temperature overnight. The mixture was filtered through celite, rinsed with THF and the solvent evaporated in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM solution (0/100-3/97)). The desired fractions were collected and concentrated in vacuo to yield rac-2-amino-2- (3-bromo-phenyl) -propan-1-ol as an oil (5.70 g, 85% yield).
Examples A4
(R)-2- Amino group -2-(3- Bromine compound - Phenyl radical )- C3 -1- Preparation of alcohols
Moving phase (CO) by ^ Daicel AD x 250mm in Chiralpak ^ Daicel ^ moving phase2MeOH with 0.2% iPrNH2) Preparation of SFC as described above, a sample of rac-2-amino-2- (3-bromo-phenyl) -propan-1-ol (15.4 g) was separated into the corresponding enantiomer to give (R) -2-amino-2- (3-bromo-phenyl) -propan-1-ol1-ol (7.21 g, 40% yield).
：-14.9 º (589 nm, c = 0.2946 w/v %, MeOH, 20 ℃).
Examples A5
rac -[1-(3- Bromine compound - Phenyl radical )-2- Hydroxy radical -1- Methyl radical - Ethyl radical ]- Preparation of tert-butyl carbamate
Di-tert-butyl dicarbonate (4.84 g, 22.16 mmol) is added portionwise at 0 ℃ to saturated NaHCO3(15 mL) and THF (15 mL) in a stirred solution of rac-2-amino-2- (3-bromo-phenyl) -propan-1-ol (1.7 g, 7.39 mmol). The mixture was stirred at 0 ℃ for 10 minutes and at room temperature for 15 hours. The mixture was cooled in an ice/water bath and was washed with KHSO4Acidifying to pH 1-2. The organic layer was separated and the aqueous layer was further extracted with EtOAc. The combined organic layers were dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc 0/100-20/80 in DCM). The desired fractions were collected and concentrated in vacuo to yield rac- [1- (3-bromo-phenyl) -2-hydroxy-1-methyl-ethyl as a colourless oil]Tert-butyl carbamate (2.36 g, 93% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a1-a 5:
examples A6
[1-(5- Bromine compound -2- Fluorophenyl group )-2- Hydroxy radical -1- Methyl ethyl radical ] Carbamic acid ester (R) Preparation of tert-butyl ester
Prepared from 1- (5-bromo-2-fluorophenyl) ethanone.
Examples A7
4-( Trifluoromethyl radical )-1-{[2-( Trimethylsilyl group ) Ethoxy radical ] Methyl radical }-1H- Preparation of imidazoles
Sodium hydride (60% in mineral oil; 0.4 g, 10 mmol) was added portionwise to a stirred solution of 4- (trifluoromethyl) -1H-imidazole (1.15 g, 8.45 mmol) in THF (19 mL) at 0 ℃. After stirring at 0 ℃ for 30 minutes, 2- (trimethylsilyl) ethoxymethyl chloride (1.69 g, 10 mmol) was added and the reaction mixture was stirred at room temperature for 30 minutes. Water was added and the product was extracted with EtOAc. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo afforded 4- (trifluoromethyl) -1- { [2- (trimethylsilyl) ethoxy]Methyl } -1H-imidazole (2.2 g, 98% yield) was used in the next step without further purification.
Following a similar synthetic procedure as described in example a7, the following intermediates were prepared:
examples A8
4- Iodine -1-{[2-( Trimethylsilyl group ) Ethoxy radical ] Methyl radical }-1H- Imidazole and 5- iodine -1-{[2-( Trimethylsilyl group ) Ethoxy radical ] Methyl radical }-1H- Preparation of mixtures of imidazoles
Prepared from 4-iodoimidazole.
Examples A9
4-( Trifluoromethyl radical )-1-{[2-( Trimethylsilyl group ) Ethoxy radical ] Methyl radical }-1H- Imidazole -2- Preparation of formaldehyde
N-butyllithium (1.6M in hexane; 7.74 mL, 12.4 mmol) was added dropwise to the stirred 4- (trifluoromethyl) -1- { [2- (trimethylsilyl) ethoxy group at-78 ℃ under nitrogen]Methyl } -1H-imidazole (2.75 g, 10.33 mmol) in THF (76.5 mL). The mixture was stirred at-78 ℃ for 10 min, then DMF (5.74 mL, 74.1 mmol) was added. The reaction mixture was stirred at-78 ℃ for 30 minutes and at 0 ℃ for 1 hour. The mixture was diluted with water and extracted with EtOAc. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; DCM in heptane 0/100-60/40). The desired fractions were collected and concentrated in vacuo to give 4- (trifluoromethyl) -1- { [2- (trimethylsilyl) ethoxy]Methyl } -1H-imidazole-2-carbaldehyde (1.6 g, 53% yield).
Examples A10
4-( Trifluoromethyl radical )-1-{[2-( Trimethylsilyl group ) Ethoxy radical ] Methyl radical }-1H- Imidazole -2- Preparation of formaldoxime
Mixing Na2CO3(0.54 g, 5.1 mmol) was added to 4- (trifluoromethyl) -1- { [2- (trimethylsilyl) ethoxy]A stirred solution of methyl } -1H-imidazole-2-carbaldehyde (1.5 g, 5.1 mmol) and hydroxylamine hydrochloride (0.71 g, 10.2 mmol) in distilled water (10.2 mL). The mixture was stirred at 70 ℃ for 1 hour, after cooling, a precipitate formed, which was filtered and washed with additional water to give 4- (trifluoromethyl) -1- { [2- (trimethylsilyl) ethoxy-1- { [2- (trifluoromethyl) ethoxy]Methyl } -1H-imidazole-2-carbaldehyde oxime (1.5 g, 95% yield).
Following a similar synthetic procedure as described in example a10, the following intermediates were prepared:
examples A11
4- Iodine -1-{[2-( Trimethylsilyl group ) Ethoxy radical ] Methyl radical }-1H- Imidazole -2- Preparation of formaldoxime
Prepared from 4-iodo-1- { [2- (trimethylsilyl) ethoxy ] methyl } -1H-imidazole-2-carbaldehyde.
Examples A12
4-( Trifluoromethyl radical )-1-{[2-( Trimethylsilyl group ) Ethoxy radical ] Methyl radical }-1H- Imidazole -2- Preparation of nitriles
Stirring of 4- (trifluoromethyl) -1- { [2- (trimethylsilyl) ethoxy at 140 deg.C]A solution of methyl } -1H-imidazole-2-carbaldehyde oxime (1.4 g, 4.53 mmol) in acetic anhydride (16.7 mL, 176.23 mmol) for 6 hours. The solvent was concentrated in vacuo, the residue was dissolved in EtOAc and Na2CO3(saturation) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo afforded 4- (trifluoromethyl) -1- { [2- (trimethylsilyl) ethoxy]Methyl } -1H-imidazole-2-carbonitrile (1.15 g, 87% yield). The product was used without further purification in the next reaction step.
Examples A13
4-( Trifluoromethyl radical )-1H- Imidazole -2- Preparation of nitriles
Stirring the 4- (trifluoromethyl) -1- { [2- (trimethylsilyl) ethoxy group in a sealed tube at 65 deg.C]Methyl } -1H-imidazole-2-carbonitrile (1.15 g, 3.95 mmol) in tetrabutylammonium fluoride (1M in THF; 25.6 mL, 25.6 mmol) for 4H. The mixture was diluted with EtOAc and buffered K2HPO4/KH2PO4And (6) processing. The organic layer was separated and the aqueous phase was washed with additional EtOAc. Drying (MgSO)4) The combined organic layers were filtered and the solvent was evaporated in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM (0/100-2/98) followed by MeOH/DCM (0/100-1/99)). The desired fractions were collected and concentrated in vacuo to give 4- (trifluoromethyl) -1H-imidazole-2-carbonitrile (0.26 g, 41% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a7-a 13:
examples A14
4- Chlorine -1H- Imidazole -2- Preparation of nitriles
Prepared from 4- (chloro) -1H-imidazole.
Examples A15
4- Bromine compound -1H- Imidazole -2- Preparation of nitriles
Prepared from 4- (bromo) -1H-imidazole.
Examples A16
4- Iodine -1-{[2-( Trimethylsilyl group ) Ethoxy radical ] Methyl radical }-1H- Imidazole -2- Formaldehyde and 5- iodine -1-{[2-( Trimethylsilyl group ) Ethoxy radical ] Methyl radical }-1H- Imidazole -2- Preparation of a mixture of Formaldehyde
Lithium diisopropylamide (1.8M; 57.6 mL, 103.6 mmol) was added dropwise to 4-iodo-1- { [2- (trimethylsilyl) ethoxy ] under nitrogen at-78 deg.C]Methyl } -1H-imidazole and 5-iodo-1- { [2- (trimethylsilyl) ethoxy]A mixture of methyl } -1H-imidazole (28 g, 86.4 mmol) in a stirred solution of THF (640 mL). -78 deg.CThe mixture was stirred for 20min, then DMF (48 mL, 620 mmol) was added. The reaction mixture was stirred at-78 ℃ for 30 minutes and at 0 ℃ for 2 hours. The mixture was diluted with EtOAc, washed with water and brine. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo afforded 4-iodo-1- { [2- (trimethylsilyl) ethoxy]Methyl } -1H-imidazole-2-carbaldehyde and 5-iodo-1- { [2- (trimethylsilyl) ethoxy]Mixture of methyl } -1H-imidazole-2-carbaldehyde (45 g, quantitative yield).
Examples A17
4- Iodine -1H- Imidazole -2- Preparation of nitriles
4-iodo-1- { [2- (trimethylsilyl) ethoxy ] was stirred at reflux]A solution of methyl } -1H-imidazole-2-carbaldehyde oxime (20 g, 54.5 mmol) in tetrabutylammonium fluoride (1M in THF; 200.5 mL, 2.12 mol) for 6 hours. The mixture was evaporated in vacuo and the residue was dissolved in EtOAc and taken up with saturated Na2CO3Washed and the organic layers dried (MgSO)4) Filtered and the solvent evaporated in vacuo. The residue was dissolved in acetic anhydride and stirred at 65 ℃ for 4 hours. The mixture was diluted with EtOAc and buffered K2HPO4/KH2PO4And (6) processing. The organic layer was separated and the aqueous phase was washed with additional EtOAc. The combined organic layers were dried (MgSO)4) Filtered and the solvent evaporated in vacuo. The crude product was purified by flash column chromatography (silica; MeOH 0/100-5/95 in DCM). The desired fractions were collected and concentrated in vacuo to afford 4-iodo-1H-imidazole-2-carbonitrile (3 g, 25% yield).
Examples A18
1H- Imidazole -2- Preparation of nitriles
Hydroxylamine hydrochloride (7.96 g, 110 mmol) was added portionwise to a stirred suspension of 2-imidazolecarboxaldehyde (10 g, 100 mmol) in pyridine (27.85 mL) at-5 ℃. The mixture was stirred at room temperature for 2 hours. The mixture was then heated at 80 ℃ and acetic anhydride (18.7 mL, 200 mmol) was added dropwise over 40 minutes to maintain the temperature below 110 ℃. After addition, the reaction mixture was stirred at 80 ℃ for 45 minutes, then cooled to 5 ℃ and basified to pH 8 with NaOH (25%). The mixture was diluted with EtOAc, the organic layer was separated and dried (Na)2SO4) Filtered and the solvent evaporated in vacuo. The crude product was diluted with DCM and stirred for 18 hours. The solid was filtered and dried in vacuo to give 1H-imidazole-2-carbonitrile (7.5 g, 77% yield), which was used in the next reaction without further purification.
Examples A19
4- Formyl radical -N,N- Dimethyl group -1H- Imidazole -1- Preparation of sulfonamides
1, 4-diazabicyclo [2.2.2 ] at 0 deg.C]Octane (21 g, 187.33 mmol) and dimethylsulfamoyl chloride (18.4 mL, 171.72 mmol) were added to a stirred suspension of 1H-imidazole-4-carbaldehyde (15 g, 156.11 mmol) in acetonitrile (300 mL). The mixture was allowed to warm to room temperature and stirred for 18 hours. The mixture was concentrated in vacuo and the residue was diluted with water and extracted with EtOAc. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc 0/100-60/40 in DCM). Collecting the desired fractions and concentrating under vacuum to obtain 4-formazan as a cream solidAcyl radicalradical-N, N-dimethyl-1H-imidazole-1-sulfonamide (27.2 g, 86% yield).
Examples A20
4-( Difluoromethyl group )-N,N- Dimethyl group -1H- Imidazole -1- Preparation of sulfonamides
The multiple systems (pumps, valves, PFA tubes and reactor loop) of the Vaporurec R2+ R4 unit were dried with isopropanol (2 mL/min, 15 min) and anhydrous THF (0.5 mL/min, 20 min). Mixing 4-JiaAcyl radicalA solution of the group-N, N-dimethyl-1H-imidazole-1-sulfonamide (0.5 g, 2.46 mmol) in DCM was loaded into the sample loop (10 mL) on Vaporurtec R2+ R4. A solution of diethylaminosulfur trifluoride (0.65 mL, 4.92 mmol) in DCM was charged into the second sample ring (10 mL). The two sample loops were transferred into a flow of DCM, flowing at 0.110 mL/min each and mixed in the reactor at 80 ℃. The mixture was then matured in the reactor using a10 mL coil. Then directly through CaCO3The output of the coil was collected. The solution was filtered through celite and the solvent was evaporated in vacuo. The crude product was purified by flash column chromatography (silica; DCM). The desired fractions were collected and concentrated in vacuo to give 4- (difluoromethyl) -N, N-dimethyl-1H-imidazole-1-sulfonamide as a pale brown solid (0.41 g, 73% yield).
Examples A21
4-( Difluoromethyl group )-1-( Dimethyl sulfonamide )-1H- Imidazole -2- Preparation of ethyl carboxylate
Potassium bis (trimethylsilyl) amide (1M in THF; 7.66 mL, 7.66 mmol) was added dropwise to a stirred solution of 4- (difluoromethyl) -N, N-dimethyl-1H-imidazole-1-sulfonamide (1.5 g, 6.66 mmol) in THF (30 mL) at-78 deg.C under nitrogen. The mixture was stirred at-78 ℃ for 1h and ethyl cyanoformate (0.76 g, 7.66 mmol) in THF (7 ml) was added. The mixture was stirred at-78 ℃ for 1 hour and then at room temperature for 2 hours. The mixture is saturated with NH4Dilute Cl and extract with EtOAc. The organic layer was separated and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc in heptane 0/100-60/40). The desired fractions were collected and concentrated in vacuo to give ethyl 4- (difluoromethyl) -1- (dimethylsulfamoyl) -1H-imidazole-2-carboxylate as a pale yellow oil (1.3 g, 66% yield).
Examples A22
4-( Difluoromethyl group )-1H- Imidazole -2- Preparation of ethyl carboxylate
Hydrochloric acid (4M in dioxane; 35 mL, 140 mmol) was added to ethyl 4- (difluoromethyl) -1- (dimethylsulfamoyl) -1H-imidazole-2-carboxylate (5.55 g, 18.67 mmol). The mixture was stirred at 50 ℃ for 2 hours. The solvent was evaporated in vacuo. The residue is subjected to NaHCO3Diluted (saturated) and extracted with DCM. The organic layer was separated and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc 0/100-10/90 in DCM). The desired fractions were collected and concentrated in vacuo to give ethyl 4- (difluoromethyl) -1H-imidazole-2-carboxylate (2.98 g, 84% yield) as a white solid.
Examples A23
1-(5- Bromine compound -2- Fluorophenyl group )-2,2- Preparation of difluoroethanone
N-butyllithium (2.5M in hexane; 20.03 mL, 50.07 mmol) was added to a stirred solution of diisopropylamine (7.02 mL, 50.07 mmol) in THF (125.4 mL) at-70 deg.C under a nitrogen atmosphere. The mixture was stirred at-70 ℃ for 30 minutes, then 4-bromofluorobenzene (5 mL, 45.51 mmol) was added dropwise. The reaction mixture was stirred at-70 ℃ for 30 min before ethyl difluoroacetate (5.74 mL, 54.62 mmol) was added. The mixture was stirred at-70 ℃ for 1 hour and then passed over NH4Diluted with Cl (sat) and extracted with ether. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; DCM in heptane 0/100-100/0). The desired fractions were collected and concentrated in vacuo to afford 1- (5-bromo-2-fluorophenyl) -2, 2-difluoroethanone as a pale yellow oil which solidified upon standing (9.3 g, 81% yield).
Examples A24
[(1Z)-1-(5- Bromine compound -2- Fluorophenyl group )-2,2- Difluoroethylene radical ] Preparation of tert-butyl carbamate
N-boc-imino- (triphenyl) phosphorane (15.26 g, 40.43 mmol) was added to a stirred solution of 1- (5-bromo-2-fluorophenyl) -2, 2-difluoroethanone (9.3 g, 36.76 mmol) in toluene (93 mL). The mixture was stirred at 90 ℃ for 18 hours. The solvent was evaporated in vacuo and the residue dissolved in heptane. The solid was filtered off and the filtrate was evaporated in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc in heptane 0/100-5/95). The desired fractions were collected and concentrated in vacuo to give tert-butyl [ (1Z) -1- (5-bromo-2-fluorophenyl) -2, 2-difluoroethylene ] carbamate as a pale yellow oil (8.7 g, 67% yield).
Examples A25
rac-[1-(5- Bromine compound -2- Fluorophenyl group )-1-( Difluoromethyl group ) C3 -2- Alkene(s) -1- Base of ] Preparation of tert-butyl carbamate
Vinylmagnesium bromide (1M in THF; 4 mL, 4 mmol) was added to stirred [ (1Z) -1- (5-bromo-2-fluorophenyl) -2, 2-difluoroethylene at-78 ℃ under nitrogen]Tert-butyl carbamate (0.94 g, 2.67 mmol) in THF (8.9 mL). The mixture was stirred at-78 ℃ for 30 minutes and at room temperature for 18 hours. The mixture is saturated with NH4Dilute Cl and extract with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc 0/100-100/0 in DCM). The desired fractions were collected and concentrated in vacuo to yield rac- [1- (5-bromo-2-fluorophenyl) -1- (difluoromethyl) prop-2-en-1-yl) as a colorless oil]Tert-butyl carbamate (1 g, 99% yield).
Examples A26
rac-[1-(5- Bromine compound -2- Fluorophenyl group )-2,2- Difluoro (F) -1-( Hydroxy methyl radical ) Ethyl radical ] Preparation of tert-butyl carbamate
To rac- [1- (5-bromo-2-fluorophenyl) -1- (difluoromethyl) prop-2-en-1-yl at-78 deg.C]A solution of tert-butyl carbamate (3.0 g, 7.9 mmol) in a mixture of DCM (50 mL) and MeOH (112 mL) was treated with ozone while maintaining the blue color (30 min). Excess ozone was removed by bubbling oxygen through for 10 minutes. Then, sodium borohydride (0.89 g, 23.7 mmol) was added and the mixture was brought to 0 ℃. After stirring for 30 min, the mixture was poured onto HCl (1N) and extracted with ether. The organic layer was separated, washed with water and brine, and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo afforded rac- [1- (5-bromo-2-fluorophenyl) -2, 2-difluoro-1- (hydroxymethyl) ethyl as a white solid]Tert-butyl carbamate (2.99 g, 98% yield).
Examples A27
(R)-[1-(5- Amino group -2- Fluorophenyl group )-2- Hydroxy radical -1- Methyl ethyl radical ] Preparation of tert-butyl carbamate
Dimethyl sulfoxide (279.5 mL) was added to (R) [1- (5-bromo-2-fluorophenyl) -2-hydroxy-1-methylethyl]Tert-butyl carbamate (6.8 g, 19.5 mmol), sodium azide (3.17 g, 48.8 mmol), copper (I) iodide (4.65 g, 24.4 mmol) and Na2CO3(4.14 g, 39.1 mmol). The mixture was degassed with nitrogen for several minutes, then N, N' -dimethylethylenediamine (3.68 mL, 34.2 mmol) was added and the mixture was stirred at 90 ℃ for 16 hours. After cooling to room temperature, the mixture was filtered through celite and washed with water. The filtrate was diluted with EtOAc and water and carefully acidified with 1M HCl. The organic layer was separated and the aqueous phase was extracted with additional EtOAc. NH in aqueous phase through water3Basified and then washed with EtOAc. The combined organic layers were dried (MgSO)4) Filtration and evaporation of the solvent in vacuo.The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-5/95). The desired fractions were collected and concentrated in vacuo to give (R) - [1- (5-amino-2-fluorophenyl) -2-hydroxy-1-methylethyl]Tert-butyl carbamate (2.7 g, 49% yield).
Examples A28
(R)-(3-{1-[( Tert-butoxycarbonyl group ) Amino group ]-2- Hydroxy radical -1- Methyl ethyl radical }-4- Fluorophenyl group ) Preparation of urethane
Ethyl chloroformate (1 mL, 10.5 mmol) was added to (R) - [1- (5-amino-2-fluorophenyl) -2-hydroxy-1-methylethyl]T-butyl carbamate (2.7 g, 9.5 mmol) in saturated NaHCO3(60 mL) and THF (50 mL). The mixture was stirred at room temperature for 4 hours. The mixture was then diluted with EtOAc, the organic layer was separated and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo afforded (R) - (3- {1- [ (tert-butoxycarbonyl) amino)]-ethyl 2-hydroxy-1-methylethyl } -4-fluorophenyl) carbamate (3.1 g, 92% yield). The product was used without further purification in the next reaction step.
Examples A29
rac-[3-( Tert-butoxycarbonyl group )-4-(3- Bromine compound - Phenyl radical )-4- Methyl radical -[1,1,3] Oxathiazolidines -2- Preparation of oxides
A solution of rac- [1- (3-bromo-phenyl) -2-hydroxy-1-methyl-ethyl ] -carbamic acid tert-butyl ester (7.2 g, 21.8 mmol) in anhydrous acetonitrile (40 mL) cooled to-40 ℃ is added dropwise to a stirred solution of thionyl chloride (3.98 mL, 54.51 mmol) in anhydrous acetonitrile (114 mL) under a nitrogen atmosphere. The reaction mixture was stirred at-40 ℃ for 30 min before addition of pyridine (8.78 mL, 109.02 mmol). The reaction was allowed to warm to room temperature and stirred for 64 hours. The solvent was evaporated in vacuo. EtOAc was added to the residue. The solid was filtered off and the filtrate was concentrated in vacuo. The residue was treated with diethyl ether. The solid was filtered off and the filtrate was concentrated in vacuo to give rac- [3- (tert-butoxy-carbonyl) -4- (3-bromo-phenyl) -4-methyl- [1,1,3] oxathiazolidin-2-oxide as an oil (7.09 g, 86% yield) which was used in the next reaction without further purification.
Examples A30
rac-[3-( Tert-butoxycarbonyl group )-4-(3- Bromine compound - Phenyl radical )-4- Methyl radical -[1,1,3] Oxathiazolidines -2,2- Preparation of dioxides
Ruthenium (III) chloride (39 mg, 0.19 mmol) was added to rac- [3- (tert-butoxycarbonyl) -4- (3-bromo-phenyl) -4-methyl- [1,1,3] at 0 deg.C]To a solution of oxathiazolidine-2-oxide (7 g, 18.6 mmol) in acetonitrile/water (1:1) (200 mL) was added sodium periodate (5.97 g, 27.91 mmol). The reaction was allowed to warm to room temperature and stirred for 2 hours. The mixture was filtered through celite, washing with EtOAc (50 mL). Water (50 mL) and EtOAc (100 mL) were added to the filtrate. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The product was purified by flash column chromatography (silica; DCM). The desired fractions were collected and concentrated in vacuo to yield rac- [3- (tert-butoxycarbonyl) -4- (3-bromo-phenyl) -4-methyl- [1,1, 3-d as a white solid]Oxathiazolidine-2, 2-dioxide (6.66 g, 91% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a29-a 30:
examples A31
(R)-4-(5- Bromine compound -2- Fluorophenyl group )-4- Methyl radical -1,2,3- Oxathiazolidines -3- Carboxylic acid tert-butyl ester 2,2- Preparation of dioxides
Prepared from tert-butyl (R) - [1- (5-bromo-2-fluorophenyl) -2-hydroxy-1-methylethyl ] carbamate.
Examples A32
rac-4-(5- Bromine compound -2- Fluorophenyl group )-4-( Difluoromethyl group )-1,2,3- Oxathiazolidines -3- Carboxylic acid tert-butyl ester 2,2- Preparation of dioxides
Prepared from rac- [1- (5-bromo-2-fluorophenyl) -2, 2-difluoro-1- (hydroxymethyl) -ethyl ] carbamic acid tert-butyl ester.
Examples A33
(R)-4-{5-[( Ethoxycarbonyl group ) Amino group ]-2- Fluorophenyl group }-4- Methyl radical -1,2,3- Oxathiazolidines -3- Carboxylic acid tert-butyl ester 2,2- Preparation of dioxides
Prepared from ethyl (R) - (3- {1- [ (tert-butoxycarbonyl) amino ] -2-hydroxy-1-methylethyl } -4-fluorophenyl) carbamate.
Examples A34
rac-1-[1-(3- Bromine compound - Phenyl radical )1- Tert-butoxycarbonylamino group - Ethyl radical ]-1H- Imidazole -2- Preparation of ethyl carboxylate
Sodium hydride (60% in mineral oil) (199 mg, 4.97 mmol) was added to a solution of imidazole-2-carboxylic acid ethyl ester (697 mg, 4.97 mmol) in DMF (33 mL) at room temperature and the reaction mixture was stirred at room temperature for 30 min. Rac- [3- (tert-butoxycarbonyl) -4- (3-bromo-phenyl) -4-methyl- [1,1,3] oxathiazolidine-2, 2-dioxide (1.95 g, 4.97 mmol) was added and the reaction mixture was heated at 100 ℃ for 64 h. Water was added and the product was extracted with DCM. The organic layer was separated, dried, filtered and the solvent evaporated in vacuo. The crude product was purified by flash column chromatography (silica: MeOH in DCM 0/100-5/95). The desired fractions were collected and concentrated in vacuo to yield rac-1- [1- (3-bromo-phenyl) 1-tert-butoxycarbonylamino-ethyl ] -1H-imidazole-2-carboxylic acid ethyl ester as a colorless oil (1.19 g, 53% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 34:
examples A35
{1-(5- Bromine compound -2- Fluorophenyl group )-2-[2- Cyano radical -4-( Trifluoromethyl radical )-1H- Imidazole -1- Base of ]-1- Methyl ethyl radical } Carbamic acid ester (R)- Preparation of tert-butyl ester
Prepared from 4- (5-bromo-2-fluorophenyl) -4-methyl-1, 2, 3-oxathiazolidine-3-carboxylic acid (R) -tert-butyl ester 2, 2-dioxide and 4- (trifluoromethyl) -1H-imidazole-2-carbonitrile.
Examples A36
(R)-[1-(5- Bromine compound -2- Fluorophenyl group )-2-(2- Cyano radical -1H- Imidazole -1- Base of )-1- Methyl ethyl radical ] Preparation of tert-butyl carbamate
1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (7.28 mL, 48.75 mmol) was added to a stirred solution of 4- (5-bromo-2-fluorophenyl) -4-methyl-1, 2, 3-oxathiazolidine-3-carboxylic acid (R) -tert-butyl ester 2, 2-dioxide (10 g, 24.4 mmol) and 1H-imidazole-2-carbonitrile (2.61 g, 28.03 mmol) in acetonitrile (80 mL). The mixture was stirred at 90 ℃ for 18 h, then diluted with HCl (1M) and extracted with DCM. The organic layer was separated and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc 0/100-10/90 in DCM). The desired fractions were collected and concentrated in vacuo to give (R) - [1- (5-bromo-2-fluorophenyl) -2- (2-cyano-1H-imidazol-1-yl) -1-methylethyl as a viscous solid]Tert-butyl carbamate (10 g, 97% yield).
Examples A37
(R)-1-{2-(5- Bromine compound -2- Fluorophenyl group )-2-[( Tert-butoxycarbonyl group ) Amino group ] Propyl radical }-4-( Difluoromethyl group )-1H- Imidazole -2- Preparation of ethyl carboxylate
Ethyl 4- (difluoromethyl) -1H-imidazole-2-carboxylate (0.5 g, 2.63 mmol) was added to stirred (R) -4- (5-bromo-2-fluorophenyl) -4-methyl-1, 2, 3-oxathiazolidine-3-carboxylate tert-butyl 2, 2-dioxide (1.03 g, 2.5 mmol) and K at room temperature2CO3(0.36 g, 2.63 mmol) in DMF (10.5 mL). The mixture was stirred at 100 ℃ for 2 hours and then concentrated in vacuo. The residue was diluted with citric acid (sat.) and EtOAc. The mixture was stirred at room temperature for 16 hours. The organic layer was separated and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; DCM). The desired fractions were collected and concentrated in vacuo to give 1- { (R) -2- (5-bromo-2-fluorophenyl) -2- [ (tert-butoxycarbonyl) amino group as a colorless oil]Propyl } -4- (difluoromethyl) -1H-imidazole-2-carboxylic acid ethyl ester (0.66 g, 51% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 37:
examples A38
(R)-{3-[1-[( Tert-butoxycarbonyl group ) Amino group ]-2-(4- Chlorine -2- Cyano radical -1H- Imidazole -1- Base of )-1- Methyl ethyl radical ]-4- Fluorophenyl group } Preparation of urethane
Prepared from (R) -4- {5- [ (ethoxycarbonyl) amino ] -2-fluorophenyl } -4-methyl-1, 2, 3-oxathiazolidine-3-carboxylic acid tert-butyl ester 2, 2-dioxide and 4-chloro-1H-imidazole-2-carbonitrile.
Examples A39
(R)-{3-[1-[( Tert-butoxycarbonyl group ) Amino group ]-2-(4- Bromine compound -2- Cyano radical -1H- Imidazole -1- Base of )-1- Methyl ethyl radical ]-4- Fluorophenyl group } Preparation of urethane
Prepared from (R) -4- {5- [ (ethoxycarbonyl) amino ] -2-fluorophenyl } -4-methyl-1, 2, 3-oxathiazolidine-3-carboxylic acid tert-butyl ester 2, 2-dioxide and 4-bromo-1H-imidazole-2-carbonitrile.
Examples A40
Preparation of ethyl (R) - {3- [1- [ (tert-butoxycarbonyl) amino ] -2- (2-cyano-4-iodo-1H-imidazol-1-yl) -1-methylethyl ] -4-fluorophenyl } carbamate
Prepared from (R) -4- {5- [ (ethoxycarbonyl) amino ] -2-fluorophenyl } -4-methyl-1, 2, 3-oxathiazolidine-3-carboxylic acid tert-butyl ester 2, 2-dioxide and 4-iodo-1H-imidazole-2-carbonitrile.
Examples A41
rac-1-[2- Amino group -2-(3- Bromine compound - Phenyl radical )- Propyl radical ]-1H- Imidazole -2- Preparation of ethyl carboxylate
Trifluoroacetic acid (5.9 mL) was added to a solution of rac-1- [1- (3-bromo-phenyl) 1-tert-butoxycarbonylamino-ethyl ] -1H-imidazole-2-carboxylic acid ethyl ester (1.18 g, 2.61 mmol) in DCM (59 mL) and the reaction mixture was stirred at room temperature overnight. The reaction was concentrated in vacuo to give rac-1- [ 2-amino-2- (3-bromo-phenyl) -propyl ] -1H-imidazole-2-carboxylic acid ethyl ester as an oil (1.49 g, 92% yield). The product was used in the next step without further purification.
Examples A42
1-[(R)-2- Amino group -2-(5- Bromine compound -2- Fluorophenyl group ) Propyl radical ]-4-( Difluoromethyl group )-1H- Imidazole -2- Preparation of ethyl carboxylate
Hydrochloric acid (4M in dioxane; 3.4 mL, 13.5 mmol) was added to 1- { (R) -2- (5-bromo-2-fluorophenyl) -2- [ (tert-butoxycarbonyl) amino]Propyl } -4- (difluoromethyl) -1H-imidazole-2-carboxylic acid ethyl ester (0.62 g, 1.19 mmol). The mixture was stirred at room temperature for 90 minutes. The solvent was evaporated in vacuo. The residue was saturated NaHCO3Dilute and extract with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo afforded 1- [ (R) -2-amino-2- (5-bromo-2-fluorophenyl) propyl]-4- (difluoromethyl) -1H-imidazole-2-carboxylic acid ethyl ester (0.5 g, 100% yield), which was used in the next step without further purification.
Examples A43
rac-1-[2- Amino group -2-(5- Bromine compound -2- Fluorophenyl group )-3,3- Difluoropropyl ]-1H- Imidazole -2- Preparation of ethyl carboxylate
1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (3.35 mL, 22.4 mmol) was added to a stirred solution of rac-4- (5-bromo-2-fluorophenyl) -4- (difluoromethyl) -1,2, 3-oxathiazolidine-3-carboxylic acid tert-butyl ester 2, 2-dioxide (5 g, 11.2 mmol) and imidazole-2-carboxylic acid ethyl ester (2.36 g, 16.8 mmol) in toluene (50 mL). The mixture was stirred at 60 ℃ for 18 hours, then the solvent was concentrated in vacuo. The residue was dissolved in THF (50 mL), treated with HCl (1N), and stirred at room temperature for 18 hours. The mixture was diluted with EtOAc, the organic layer was separated and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo gave a residue which was dissolved in 4M HCl/dioxane (28 mL) and stirred at room temperature for 1 hour. The mixture was evaporated in vacuo, the residue was suspended in DCM and saturated NaHCO3And (6) washing. The organic layer was separated and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo gave rac-1- [ 2-amino-2- (5-bromo-2-fluorophenyl) -3, 3-difluoropropyl as a white solid]-1H-imidazole-2-carboxylic acid ethyl ester (3.95 g, 87% yield). The product was used without further purification in the next reaction step.
Examples A44
(R)-[1-(5- Bromine compound -2- Fluorophenyl group )-2-(4- Chlorine -2- Cyano radical -1H- Imidazole -1- Base of )-1- Methyl ethyl radical ] Preparation of tert-butyl carbamate
1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (10.3 mL, 68.8 mmol) was added to a stirred solution of (R) -4- (5-bromo-2-fluorophenyl) -4-methyl-1, 2, 3-oxathiazolidine-3-carboxylic acid tert-butyl ester 2, 2-dioxide (28.24 g, 34.41 mmol) and 4-chloro-1H-imidazole-2-carbonitrile (7.9 g, 61.94 mmol) in acetonitrile (203 mL). The mixture was stirred at 100 ℃ for 2 h, then diluted with DCM and washed with HCl (1N). The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc 0/100-10/90 in DCM). The desired fractions were collected and evaporated in vacuo to give (R) - [1- (5-bromo-2-fluorophenyl) -2- (4-chloro-2-cyano-1H-imidazol-1-yl) -1-methylethyl as a white solid]Tert-butyl carbamate (15.5 g, 98% yield).
Examples A45
(R)-1-[2- Amino group -2-(3- Bromine compound - Phenyl radical )- Propyl radical ]-1H- Imidazole -2- Preparation of ethyl carboxylate
Cesium carbonate (4.98 g, 15.3 mmol) was added to imidazole-2-carboxylic acid ethyl ester (1.39 g, 9.94 mmol) and (R) - [3- (tert-butoxycarbonyl) -4- (3-bromo-phenyl) -4-methyl- [1,1,3]Oxathiazolidine-2, 2-dioxide (3 g, 7.65 mmol) in a mixture of anhydrous acetonitrile (36 mL). The mixture was stirred at 130 ℃ for 45 minutes under microwave irradiation. Then NH is added4Cl (saturated) and product extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. HCl (4M in dioxane) was then added and the mixture was stirred at room temperature for 1 hour. The solvent was concentrated in vacuo, the residue was suspended in DCM and NaHCO3Saturation ofAnd (4) washing the solution. The organic layer was separated and dried (MgSO)4) Filtered and the solvent evaporated in vacuo. The crude product was purified by flash column chromatography (silica: EtOAc). The desired fractions were collected and concentrated in vacuo to give (R) -1- [ 2-amino-2- (3-bromo-phenyl) -propyl ] as a white solid]-1H-imidazole-2-carboxylic acid ethyl ester (1.46g, 54% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 45:
examples A46
(R)-1-[2- Amino group -2-(5- Bromine compound -2- Fluorophenyl group ) Propyl radical ]-1H- Imidazole -2- Preparation of ethyl carboxylate
Prepared from (R) -4- (5-bromo-2-fluorophenyl) -4-methyl-1, 2, 3-oxathiazolidine-3-carboxylic acid tert-butyl ester 2, 2-dioxide.
Examples A47
rac-6-(3- Bromine compound - Phenyl radical )-6- Methyl radical -6,7- Dihydro -5H- Imidazo [1,2-a] Pyrazine esters -8- Preparation of ketones
Heating rac-1- [ 2-amino-2- (3-bromo-phenyl) -propyl ] at 90 deg.C]-1H-imidazole-2-carboxylic acid ethyl ester (1.4 g, 3 mmol) in ethanol (10 mL) for 24H. The reaction mixture was concentrated in vacuo and saturated NaHCO3Work up and extract the product with DCM. The organic layer was separated and dried (MgSO)4)，Filtration and evaporation of the solvent in vacuo afforded rac-6- (3-bromo-phenyl) -6-methyl-6, 7-dihydro-5H-imidazo [1,2-a ] as a white solid]Pyrazin-8-one (0.6 g, 65% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 47:
examples A48
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-6- Methyl radical -6,7- Dihydroimidazo compounds [1,2-a]- Pyrazine esters -8(5H)- Preparation of ketones
Prepared from (R) -1- [ 2-amino-2- (5-bromo-2-fluorophenyl) propyl ] -1H-imidazole-2-carboxylic acid ethyl ester.
Examples A49
rac-6-(5- Bromine compound -2- Fluorophenyl group )-6-( Difluoromethyl group )-6,7- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8(5H)- Preparation of ketones
Trimethylaluminum 2M in toluene (10.6 mL, 21.1 mmol) was added to stirred rac-1- [ 2-amino-2- (5-bromo-2-fluorophenyl) -3, 3-difluoropropyl at 0 deg.C]-1H-imidazole-2-carboxylic acid ethyl ester (3.9 g, 9.6 mmol) in THF (39 mL). The mixture was stirred for 30 minutes at 120 ℃ under microwave irradiation. The mixture was heated over Na at 0 deg.C2CO3Diluted (saturated) and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. CoarseThe product was purified by flash column chromatography (silica; EtOAc). The desired fractions were collected and concentrated in vacuo to afford rac-6- (5-bromo-2-fluorophenyl) -6- (difluoromethyl) -6, 7-dihydroimidazo [1,2-a ] as a white solid]Pyrazin-8 (5H) -one (3 g, 87% yield).
Examples A50
(R)-6-(3- Bromine compound - Phenyl radical )-6- Methyl radical -6,7- Dihydro -5H- Imidazo [1,2-a]- Pyrazine esters -8- Preparation of ketones
Sodium methoxide (25% in MeOH) (1.9 mL, 8.29 mmol) was added to (R) -1- [ 2-amino-2- (3-bromo-phenyl) -propyl]-1H-imidazole-2-carboxylic acid ethyl ester (1.46g, 4.15 mmol) in MeOH (5 mL) and the reaction mixture heated at 55 deg.C for 18H. Concentrate the reaction in vacuo, saturated NH4Treated with Cl and the product extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo gave (R) -6- (3-bromo-phenyl) -6-methyl-6, 7-dihydro-5H-imidazo [1,2-a ] as a white solid]Pyrazin-8-one (1.2 g, 95% yield).
Examples A51
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-2-( Difluoromethyl group )-6- Methyl radical -6,7- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8(5H)- Preparation of ketones
Will K2CO3(1.32 g, 13.42 mmol)To the stirred 1- [ (R) -2-amino-2- (5-bromo-2-fluorophenyl) propyl group]-4- (difluoromethyl) -1H-imidazole-2-carboxylic acid ethyl ester (3.6 g, 7.45 mmol) in EtOH (51.1 mL) and the reaction mixture was heated at 80 ℃ for 4H. The mixture was concentrated in vacuo over NH4Cl (sat) and the product was treated with EtOAc. The organic layer was separated and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo gave (R) -6- (5-bromo-2-fluorophenyl) -2- (difluoromethyl) -6-methyl-6, 7-dihydroimidazo [1,2-a ] as a white foam]Pyrazin-8 (5H) -one (2.89 g, 100% yield). The product was used in the next reaction without further purification.
Examples A52
rac-6-(3- Bromine compound - Phenyl radical )-6- Methyl radical -6,7- Dihydro -5H- Imidazo [1,2-a]- Pyrazine esters -8- Preparation of thioketones
Phosphorus pentasulfide (0.65 g, 2.94 mmol) was added to a solution of rac-6- (3-bromo-phenyl) -6-methyl-6, 7-dihydro-5H-imidazo [1,2-a ] pyrazin-8-one (0.6 g, 1.96 mmol) in pyridine (7 mL) and the mixture was heated at 95 ℃ for 18H. The solvent was evaporated in vacuo and the residue was purified by flash column chromatography (silica gel; EtOAc 0/100-100/0 in DCM). The desired fractions were collected and concentrated in vacuo to afford rac-6- (3-bromo-phenyl) -6-methyl-6, 7-dihydro-5H-imidazo [1,2-a ] pyrazine-8-thione as a yellow solid (0.49 g, 78% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 52:
examples A53
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-6- Methyl radical -6,7- Dihydroimidazo compounds [1,2-a]- Pyrazine esters -8(5H)- Preparation of thioketones
Prepared from (R) -6- (5-bromo-2-fluorophenyl) -6-methyl-6, 7-dihydroimidazo [1,2-a ] pyrazin-8 (5H) -one.
Examples A54
rac-6-(5- Bromine compound -2- Fluorophenyl group )-6-( Difluoromethyl group )-6,7- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8(5H)- Preparation of thioketones
Prepared from rac-6- (5-bromo-2-fluorophenyl) -6- (difluoromethyl) -6, 7-dihydroimidazo [1,2-a ] -pyrazin-8 (5H) -one.
Examples A55
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-2-( Difluoromethyl group )-6- Methyl radical -6,7- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8(5H)- Preparation of thioketones
Prepared from (R) -6- (5-bromo-2-fluorophenyl) -2- (difluoromethyl) -6-methyl-6, 7-dihydroimidazo [1,2-a ] pyrazin-8 (5H) -one.
Examples A56
rac-6-(3- Bromine compound - Phenyl radical )-6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -8- Preparation of amines
32% aqueous ammonia solution (3.2 mL, 54.7 mmol) was added to stirred (rac-6- (3-bromo-phenyl) -6-methyl-6, 7-dihydro-5H-imidazo [1,2-a ] in a sealed tube]Pyrazine-8-thione (049 g, 1.52 mmol) in 7N ammonia in MeOH (3.3 mL, 22.8 mmol). The mixture was stirred at 60 ℃ for 96 hours. After cooling to room temperature, the mixture was diluted with water and Na2CO3(saturated aqueous solution) and extracted with DCM. The organic layer was separated and dried (Na)2SO4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica gel; 7M ammonia/MeOH/DCM solution 0/100-2/98 to 3/97-10/90). The desired fractions were collected and concentrated in vacuo to afford rac-6- (3-bromo-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a yellow solid]Pyrazin-8-ylamine (0.44g, 95% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 56:
examples A57
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a]- Pyrazine esters -8- Preparation of amines
Prepared from (R) -6- (5-bromo-2-fluorophenyl) -6-methyl-6, 7-dihydroimidazo [1,2-a ] pyrazine-8 (5H) -thione.
Examples A58
rac-6-(5- Bromine compound -2- Fluorophenyl group )-6-( Difluoromethyl group )-5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Ammonium chloride (0.91 g, 17 mmol) was added to rac-6- (5-bromo-2-fluorophenyl) -6- (difluoromethyl) -6, 7-dihydroimidazo [1,2-a ] in a sealed tube]Pyrazine-8 (5H) -thione (1.6 g, 4.25 mmol) in a stirred solution of 7N ammonia in MeOH (32 mL, 63.8 mmol). The mixture was stirred at 80 ℃ for 18 hours. The mixture was concentrated in vacuo, the residue suspended in DCM and washed with water. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-2/98). The desired fractions were collected and concentrated in vacuo to afford rac-6- (5-bromo-2-fluorophenyl) -6- (difluoromethyl) -5, 6-dihydroimidazo [1,2-a ] as a white solid]Pyrazin-8-amine (1.5 g, 98% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 58:
examples A59
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-2-( Difluoromethyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Prepared from (R) -6- (5-bromo-2-fluorophenyl) -2- (difluoromethyl) -6-methyl-6, 7-dihydroimidazo [1,2-a ] pyrazine-8 (5H) -thione.
Examples A60
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-6- Methyl radical -2-( Trifluoromethyl radical )-5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
(R) - {1- (5-bromo-2-fluorophenyl) -2- [ 2-cyano-4- (trifluoromethyl) -1H-imidazol-1-yl ] was stirred at 70 deg.C]A solution of tert-butyl-1-methylethyl } carbamate (2.95 g, 6 mmol) in HCl 4M/dioxane (50 mL) for 2 hours. The mixture was concentrated in vacuo, the residue suspended in DCM and Na2CO3(saturation) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-1/99). The desired fractions were collected and evaporated in vacuo to give (R) -6- (5-bromo-2-fluorophenyl) -6-methyl-2- (trifluoromethyl) -5, 6-dihydroimidazo [1,2-a]Pyrazin-8-amine (1.5 g, 64% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 60:
examples A61
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a]- Pyrazine esters -8- Preparation of amines
Prepared from tert-butyl (R) - [1- (5-bromo-2-fluorophenyl) -2- (2-cyano-1H-imidazol-1-yl) -1-methylethyl ] carbamate.
Examples A62
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-2- Chlorine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Prepared from tert-butyl (R) - [1- (5-bromo-2-fluorophenyl) -2- (4-chloro-2-cyano-1H-imidazol-1-yl) -1-methylethyl ] carbamate.
Examples A63
(R)-{3-[8- Amino group -2- Chlorine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a]- Pyrazine esters -6- Base of ]-4- Fluorophenyl group } Preparation of urethane
Prepared from ethyl (R) - {3- [1- [ (tert-butoxycarbonyl) amino ] -2- (4-chloro-2-cyano-1H-imidazol-1-yl) -1-methylethyl ] -4-fluorophenyl } carbamate.
Examples A64
(R)-{3-[8- Amino group -2- Bromine compound -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a]- Pyrazine esters -6- Base of ]-4- Fluorophenyl group } Preparation of urethane
Prepared from ethyl (R) - (3- {2- (4-bromo-2-cyano-1H-imidazol-1-yl) -1- [ (tert-butoxycarbonyl) amino ] -1-methylethyl } -4-fluorophenyl) carbamate.
Examples A65
(R)-{3-[8- Amino group -2- Iodine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a]- Pyrazine esters -6- Base of ]-4- Fluorophenyl group } Preparation of urethane
Prepared from ethyl (R) - {3- [1- [ (tert-butoxycarbonyl) amino ] -2- (2-cyano-4-iodo-1H-imidazol-1-yl) -1-methylethyl ] -4-fluorophenyl } carbamate.
Examples A66
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-3- Chlorine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
N-Chlorosuccinimide (0.123 g, 0.92 mmol) was added to stirred (R) -6- (5-bromo-2-fluorophenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a [)]Pyrazin-8-amine (0.27 g, 0.84 mmol) in acetic acid (6.23 mL). The mixture was stirred at 80 ℃ for 16 h, then diluted with water and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-3/97). The desired fractions were collected and evaporated in vacuo to give (R) -6- (5-bromo-2-fluorophenyl) -3-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] as a white solid]Pyrazin-8-amine (0.18 g, 61% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 66:
examples A67
(R)-6-(5- Bromine compound -2- Fluorophenyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a]- Pyrazine esters -8- Preparation of amines
Prepared from (R) -6- (5-bromo-2-fluorophenyl) -2-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] -pyrazin-8-amine.
Examples A68
(R)-6-(5- Amino group -2- Fluorophenyl group )-2- Chlorine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
(R) - {3- [ 8-amino-2-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] is stirred at 110 deg.C]Pyrazin-6-yl]A mixture of ethyl-4-fluorophenyl } carbamate (0.2 g, 0.55 mmol), sulfuric acid (2 mL, 37.5 mmol), water (2 mL), and acetic acid (0.8 mL, 13.9 mmol) for 2 hours. The mixture was dissolved in EtOAc and saturated Na2CO3Alkalizing. The organic layer was separated and dried (MgSO)4) Filtering and vacuum evaporating solvent to obtain (R) -6- (5-amino-2-fluorophenyl) -2-chloro-6-methyl-5, 6-dihydroimidazo [1, 2-a)]Pyrazin-8-amine (0.17 g, 79% yield), which was used in the next step without any further purification.
Examples A69
(R)-6-(5- Amino group -2- Fluorophenyl group )-2- Bromine compound -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Potassium hydroxide (0.98 g, 17.6 mmol) was added to the stirred (R) - {3- [ 8-amino-2-bromo-6-methyl-5, 6-dihydroimidazo [1,2-a ]]Pyrazin-6-yl]Ethyl-4-fluorophenyl } carbamate (0.72 g, 1.76 mmol) in EtOH (10.25 mL). The mixture was stirred at 85 ℃ for 24 hours. The mixture was diluted with DCM and EtOAc and washed with water. The organic layer was separated and dried (MgSO)4) Filtering and vacuum evaporating solvent to obtain (R) -6- (5-amino-2-fluorophenyl) -2-bromo-6-methyl-5, 6-dihydroimidazo [1, 2-a)]Pyrazin-8-amine (0.6 g, 100% yield), which was used in the next step without any further purification.
The following intermediates were prepared according to a similar synthetic procedure described in example a 69:
examples A70
(R)-6-(5- Amino group -2- Fluorophenyl group )-2- Iodine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Prepared from ethyl (R) - {3- [ 8-amino-2-iodo-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-6-yl ] -4-fluorophenyl } carbamate.
Examples A71
rac-6-(3- Amino group - Phenyl radical )-6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -8- Preparation of amines
A solution of rac-6- (3-nitro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] pyrazin-8-ylamine (310 mg, 1.14 mmol) in ethanol (28 mL) was hydrogenated in an H-Cube reactor (1 mL/min., 30mm, Pd/C5% cartridge, perhydro mode, room temperature, 2 cycles). The reaction was concentrated in vacuo to give rac-6- (3-amino-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] pyrazin-8-ylamine as a white solid (250 mg, 91%).
Examples A72
(R)-6-[5-( Diphenylmethylene - Amino group )-2- Fluorine - Phenyl radical ]-6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -8- Preparation of amines
Toluene (10 mL) was added to (R) -6- (5-bromo-2-fluoro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] under nitrogen at room temperature in a sealed tube]Pyrazin-8-ylamine (0.60 g, 1.86 mmol), tris (dibenzylideneacetone) dipalladium (0) (0.17 g, 0.19 mmol), rac-2,2 '-bis (diphenylphosphino) -1,1' -binaphthyl (0.35 g, 0.56 mmol) and sodium tert-butoxide (0.32 g, 3.3 mmol). The mixture was purged with nitrogen for several minutes, and then benzophenone imine (0.62 mL, 3.71 mmol) was added and the mixture was stirred at 100 ℃ for 2 hours. After cooling to room temperature, the mixture was diluted with water and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 0/100-2.5/96.5 in 7M ammonia/MeOH/DCM). The desired fractions were collected and concentrated in vacuo to give (R) -6- [5- (diphenylmethylene-amino) -2-fluoro-phenyl) as a yellow solid]-6-methyl-5, 6-dihydro-imidazo [1,2-a [ ]]Pyrazin-8-ylamine (0.55 g, 70% yield).
Examples A73
(R)-6-(5- Amino group -2- Fluorine - Phenyl radical )-6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a]- Pyrazine esters -8- Preparation of amines
H is to be237% hydrochloric acid in O (0.11 mL) was added to (R) -6- [5- (diphenylmethylene-amino) -2-fluoro-phenyl]-6-methyl-5, 6-dihydro-imidazo [1,2-a [ ]]Pyrazin-8-ylamine (0.31 g, 0.73 mmol) in isopropanol (8 mL). Stirring the mixture at room temperatureFor 2 hours. The solvent was evaporated in vacuo. Diethyl ether was added to the residue, and the mixture was stirred at room temperature for 15 minutes. The precipitate is filtered off, washed with diethyl ether and dried in vacuo. The residue was suspended in DCM and NaHCO3(saturation) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo afforded (R) -6- (5-amino-2-fluoro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a]Pyrazin-8-ylamine (0.11 g, 58% yield), which was used in the next step without any further purification.
The following intermediates were prepared according to a similar synthetic procedure described in example a72-a 73:
examples A74
rac-6-(5- Amino group -2- Fluorophenyl group )-6-( Difluoromethyl group )-5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Prepared from rac-6- (5-bromo-2-fluorophenyl) -6- (difluoromethyl) -5, 6-dihydroimidazo [1,2-a ] -pyrazin-8-amine.
Examples A75
(R)-6-(5- Amino group -2- Fluorophenyl group )-2-( Difluoromethyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Prepared from (R) -6- (5-bromo-2-fluorophenyl) -2- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-amine.
Examples A76
(R)-6-(5- Amino group -2- Fluorophenyl group )-6- Methyl radical -2-( Trifluoromethyl radical )-5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Dimethyl sulfoxide (69.5 mL) was added to (R) -6- (5-bromo-2-fluorophenyl) -6-methyl-2- (trifluoromethyl) -5, 6-dihydroimidazo [1,2-a]Pyrazin-8-amine (1.9 g, 4.86 mmol), sodium azide (0.79 g, 12.1 mmol), copper (I) iodide (1.16 g, 6.1 mmol) and Na2CO3(1.03 g, 9.7 mmol). The mixture was degassed over nitrogen for several minutes, then N, N' -dimethylethylenediamine (0.91 mL, 8.5 mmol) was added and the mixture was stirred at 110 ℃ for 3 hours. After cooling to room temperature, the mixture was diluted with DCM and NH4OH (32%) wash. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-3/97). The desired fractions were collected and concentrated in vacuo to give (R) -6- (5-amino-2-fluorophenyl) -6-methyl-2- (trifluoromethyl) -5, 6-dihydroimidazo [1,2-a]Pyrazin-8-amine (0.72 g, 45% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 76:
examples A77
(R)-6-(5- Amino group -2- Fluorophenyl group )-2-( Difluoromethyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Prepared from (R) -6- (5-bromo-2-fluorophenyl) -2- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-amine.
Examples A78
(R)-6-(5- Amino group -2- Fluorophenyl group )-3- Chlorine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Prepared from (R) -6- (5-bromo-2-fluorophenyl) -3-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] -pyrazin-8-amine.
Examples A79
(R)-6-(5- Amino group -2- Fluorophenyl group )-2,3- Dichloro (phenyl) methane -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Prepared from (R) -6- (5-bromo-2-fluorophenyl) -2, 3-dichloro-6-methyl-5, 6-dihydroimidazo- [1,2-a ] pyrazin-8-amine.
Examples A80
(R)-[2- Bromine compound -6-(2- Fluorine -5-{[(5- Fluoropyridines -2- Base of ) Carbonyl radical ] Amino group } Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Di-tert-butyl dicarbonate (0.060 g, 0.27 mmol) is added to the stirred (R) -N- {3- [ 8-amino-2-bromo-6-methyl-5, 6-dihydroimidazo [1,2-a ] at 0 deg.C]Pyrazin-6-yl]-4-fluorophenyl } -5-fluoropyridine-2-carboxamide (0.07 g, 0.15 mmol) in saturated NaHCO3(1 mL) and THF (2 mL). The mixture was stirred at room temperature for 18 hours. The organic layer was separated, further extracted with EtOAc and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo afforded (R) - [ 2-bromo-6- (2-fluoro-5- { [ (5-fluoropyridin-2-yl) carbonyl]Amino } phenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-8-yl radical]Tert-butyl carbamate (0.085 g, 100% yield) was used in the next step without any further purification.
Examples A81
(R)-(3-{8-[( Tert-butoxycarbonyl group ) Amino group ]-2- Chlorine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -6- Base of }-4- Fluorophenyl group ) Preparation of tert-butyl carbamate
Di-tert-butyl dicarbonate (2.89 g, 13.28 mmol) is added to the stirred (R) -6- (5-amino-2-fluorophenyl) -2-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a ]]Pyrazin-8-amine (1.3 g)4.43 mmol) and DIPEA (1.91 mL, 11.06 mL) in DCM (77.5 mL). The mixture was stirred at room temperature for 24 hours. The organic layer was diluted with DCM and saturated NaHCO3And (6) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc 0/100-30/70 in DCM). The desired fractions were collected and evaporated in vacuo to give (R) - (3- {8- [ (tert-butoxycarbonyl) amino)]-2-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-6-yl } -4-fluorophenyl) carbamic acid tert-butyl ester (1.5 g, 69% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 81:
examples A82
(R)-[6-{5-[( Tert-butoxycarbonyl group ) Amino group ]-2- Fluorophenyl group }-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from (R) -6- (5-amino-2-fluorophenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-amine.
Examples A83
(R)-(3-{8-[( Tert-butoxycarbonyl group ) Amino group ]-2- Chlorine -3- Iodine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -6- Base of }-4- Fluorophenyl group ) Preparation of tert-butyl carbamate
N-butyllithium (2.7M in heptane; 11.92 mL, 32.2 mmol) was added dropwise to the stirred (R) - (3- {8- [ (tert-butoxycarbonyl) amino group at-78 deg.C under nitrogen]-2-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-6-yl } -4-fluorophenyl) carbamic acid tert-butyl ester (1.2 g, 2.43 mmol) in THF (12 mL). The mixture was stirred at-78 ℃ for 15 minutes. A solution of iodine (11.1 g, 43.73 mmol) in THF (20 mL) was then added at-78 deg.C. The mixture was stirred at-78 ℃ for 10 minutes. The mixture was then brought to room temperature, diluted with EtOAc, then water and Na2S2O3And (6) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-5/95). The desired fractions were collected and evaporated in vacuo to give (R) - (3- {8- [ (tert-butoxycarbonyl) amino)]-2-chloro-3-iodo-6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-6-yl } -4-fluorophenyl) carbamic acid tert-butyl ester (1.46g, 97% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 83:
examples A84
(R)-[6-{5-[( Tert-butoxycarbonyl group ) Amino group ]-2- Fluorophenyl group }-3- Iodine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
From (A), (B)R)-[6-{5-[( Tert-butoxycarbonyl group ) Amino group ]-2- Fluorophenyl group }-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate.
Examples A85
Preparation of tert-butyl (R) - [6- {5- [ (tert-butoxycarbonyl) amino ] -2-fluorophenyl } -3- (hydroxymethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate
N-butyllithium (2.7M in heptane; 25 mL, 67.5 mmol) was added dropwise to the stirred (R) - [6- {5- [ (tert-butoxycarbonyl) amino group at-78 deg.C under nitrogen]-2-fluorophenyl } -6-methyl-5, 6-dihydroimidazo [1,2-a [ ]]Pyrazin-8-yl radical]Tert-butyl carbamate (3.1 g, 6.75 mmol) in THF (110 mL). The mixture was stirred at-78 ℃ for 10 minutes. Then, at-78 ℃, paraformaldehyde (6 g) was added. The mixture was then brought to room temperature, diluted with EtOAc and washed with water. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-5/95). The desired fractions were collected and evaporated in vacuo to yield (R) - [6- {5- [ (tert-butoxycarbonyl) -amino ] -ethyl acetate]-2-fluorophenyl } -3- (hydroxymethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]-pyrazin-8-yl]Tert-butyl carbamate (1.83 g, 55% yield).
Examples A86
Preparation of tert-butyl (R) - [6- {5- [ (tert-butoxycarbonyl) amino ] -2-fluorophenyl } -3-formyl-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate
Manganese dioxide (3.75 g, 43.1 mmol) was added to a stirred solution of tert-butyl (R) - [6- {5- [ (tert-butoxycarbonyl) amino ] -2-fluorophenyl } -3- (hydroxymethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate (1.63 g, 3.33 mmol) in DCM (12 mL). The mixture was stirred at room temperature for 2 hours. The mixture was filtered through celite and the filtrate evaporated in vacuo to give tert-butyl (R) - [6- {5- [ (tert-butoxycarbonyl) amino ] -2-fluorophenyl } -3-formyl-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate (1.42 g, 87% yield) which was used in the next step without further purification.
Examples A87
Preparation of tert-butyl (R) - {3- [8- [ (tert-butoxycarbonyl) amino ] -3- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-6-yl ] -4-fluorophenyl } carbamate
DAST (0.78 mL, 6.41 mmol) was added dropwise to stirred (R) - [6- {5- [ (tert-butoxycarbonyl) amino at 0 deg.C]-2-fluorophenyl } -3-formyl-6-methyl-5, 6-dihydroimidazo [1,2-a [ ]]Pyrazin-8-yl radical]Tert-butyl carbamate (1.42 g, 2.91 mmol) in DCM (15 mL). The mixture was stirred at room temperature for 16 hours. The mixture was evaporated in vacuo. The residue was dissolved in DCM and saturated NaHCO3Alkalizing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-1/99). The desired fractions were collected and evaporated in vacuo to give (R) - {3- [8- [ (tert-butoxycarbonyl) amino]-3- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-6-yl]Tert-butyl-4-fluorophenyl } carbamate (0.44g, 29% yield).
Examples A88
Preparation of tert-butyl (R) - [6- {5- [ (tert-butoxycarbonyl) amino ] -2-fluorophenyl } -2-iodo-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate
Di-tert-butyl dicarbonate (2.27 g, 10.38 mmol) is added to a stirred solution of (R) -6- (5-amino-2-fluorophenyl) -2-iodo-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-amine (2 g, 5.19 mmol) in DCM (50 mL). The mixture was stirred at room temperature for 1 hour. The solvent was evaporated in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-2/98). The desired fractions were collected and evaporated in vacuo to give tert-butyl (R) - [6- {5- [ (tert-butoxycarbonyl) amino ] -2-fluorophenyl } -2-iodo-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate (1 g, 33% yield).
Examples A89
(R)-{3-[8- Amino group -2- Chlorine -3- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -6- Base of ]-4- Fluorophenyl group } Preparation of tert-butyl carbamate
Tris (dibenzylideneacetone) dipalladium (0) (38.4 mg, 0.042 mmol) was added to the stirred (R) - (3- {8- [ (tert-butoxycarbonyl) amino group]-2-chloro-3-iodo-6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-6-yl } -4-fluorophenyl) carbamic acid tert-butyl ester (1.3 g, 2.1 mmol), 1' -bis (diphenylphosphino) ferrocene (46.5 mg, 0.084 mmol), zinc (16.5 mg, 0.25 mmol) and zinc cyanide (0.492 g, 4.19 mmol) in suspension in DMA (12 mL). The mixture was stirred for 30 minutes at 150 ℃ under microwave irradiation. The mixture was filtered through celite, the residue diluted with DCM and NH4And (5) OH washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; MeOH in DCM 0/100-3/97). The desired fractions were collected and concentrated in vacuo to give (R) - {3- [ 8-amino-2-chloro-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ]]Pyrazin-6-yl]Tert-butyl-4-fluorophenyl } carbamate (0.56 g, 64% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 89:
examples A90
(R)-{3-[8- Amino group -2- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds -[1,2-a] Pyrazine esters -6- Base of ]-4- Fluorophenyl group } Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- {5- [ (tert-butoxycarbonyl) amino ] -2-fluorophenyl } -2-iodo-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate.
Examples A91
(R)-{3-[8- Amino group -3- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds -[1,2-a] Pyrazine esters -6- Base of ]-4- Fluorophenyl group } Preparation of tert-butyl carbamate
Tetrakis (triphenylphosphine) palladium (0) (0.24 g, 0.2 mmol) was added to a stirred solution of tert-butyl (R) - (3- {8- [ (tert-butoxycarbonyl) amino ] -3-iodo-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-6-yl } -4-fluorophenyl) carbamate (1.2 g, 2.05 mmol) and zinc cyanide (1.93 g, 16.4 mmol) in DMF (6 mL). The mixture was stirred at 160 ℃ for 10 minutes under microwave irradiation. The mixture was filtered through celite and the solvent was evaporated in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-3/97). The desired fractions were collected and concentrated in vacuo to give tert-butyl (R) - {3- [ 8-amino-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-6-yl ] -4-fluorophenyl } carbamate (0.4 g, 51% yield).
Examples A92
(R)-8- Amino group -6-(5- Amino group -2- Fluorophenyl group )-2- Chlorine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -3- Preparation of nitriles
Trifluoroacetic acid (5 mL, 65.34 mmol) was added to stirred (R) - {3- [ 8-amino-2-chloro-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ]]Pyrazin-6-yl]Tert-butyl-4-fluorophenyl } carbamate (0.56 g, 1.34 mmol) in DCM (20 mL). The mixture was stirred at room temperature for 30 minutes. The solvent was evaporated in vacuo and the residue dissolved in DCM and saturated NaHCO3And (6) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; MeOH in DCM 0/100-3/97). The desired fractions were collected and concentrated in vacuo to give (R) -8-amino-6- (5-amino-2-fluorophenyl) -2-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazine-3-carbonitrile (0.21 g, 49% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 92:
examples A93
(R)-8- Amino group -6-(5- Amino group -2- Fluorophenyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -3- Preparation of nitriles
Prepared from tert-butyl (R) - {3- [ 8-amino-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-6-yl ] -4-fluorophenyl } carbamate.
Examples A94
(R)-6-(5- Amino group -2- Fluorophenyl group )-3-( Difluoromethyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Preparation of amines
Prepared from tert-butyl (R) - {3- [8- [ (tert-butoxycarbonyl) amino ] -3- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-6-yl ] -4-fluorophenyl } carbamate.
Examples A95
(R)-8- Amino group -6-(5- Amino group -2- Fluorophenyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -2- Preparation of nitriles
Prepared from tert-butyl (R) - {3- [ 8-amino-2-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-6-yl ] -4-fluorophenyl } carbamate.
Examples A96
(R)-[6-(5- Amino group -2- Fluorophenyl group )-2- Chlorine -3- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Di-tert-butyl dicarbonate (0.14 g, 0.65 mmol) is added to the stirred (R) -8-amino-6- (5-amino-2-fluorophenyl) -2-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a []Pyrazine-3-carbonitrile (0.21 g, 0.66 mmol) in DCM (11.5 mL). The mixture was stirred at room temperature for 30 minutes. The mixture was saturated NaHCO3Diluted and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; MeOH in DCM 0/100-3/97). The desired fractions were collected and evaporated in vacuo to yield (R) - [6- (5-amino-2-fluorophenyl) -2-chloro-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ]]Pyrazin-8-yl radical]Tert-butyl carbamate (0.25 g, 91% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 96:
examples A97
(R)-[6-(5- Amino group -2- Fluorophenyl group )-3- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from (R) -8-amino-6- (5-amino-2-fluorophenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazine-3-carbonitrile.
Examples A98
(R)-(3-{8-[( Tert-butoxycarbonyl group ) Amino group ]-2- Chlorine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -6- Base of }-4- Fluorophenyl group ) Preparation of tert-butyl carbamate
Prepared from (R) -6- (5-amino-2-fluorophenyl) -2, 3-dichloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-amine.
Examples A99
(R)-[6-(5- Amino group -2- Fluorophenyl group )-3-( Difluoromethyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from (R) -6- (5-amino-2-fluorophenyl) -3- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-amine.
Examples A100
(R)-[2- Chlorine -3- Cyano radical -6-(2- Fluorine -5-{[(5- Fluoropyridines -2- Base of ) Carbonyl radical ] Amino group } Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
5-fluoro-2-pyridinecarboxylic acid (15 mg, 0.11 mmol) was added to a solution of 4- (4, 6-dimethoxy-1, 3, 5-triazin-2-yl) -4-methylmorpholinium chloride (31.7 mg, 0.11 mmol) in MeOH (0.95 mL). The mixture was stirred at room temperature for 5 minutes. Cooling the mixture to 0 deg.C and adding (R) - [6- (5-amino-2-fluorophenyl) -2-chloro-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ]]Pyrazin-8-yl radical]A solution of tert-butyl carbamate (40 mg, 0.095 mmol) in MeOH (0.95 mL). The mixture was allowed to warm to room temperature and stirred for 24 hours. The solvent was evaporated in vacuo. The crude product is purified by flash column chromatography (silica; 7N NH)30/100-3/97 in MeOH/DCM). The desired fractions were collected and the solvent was evaporated in vacuo to give (R) - [ 2-chloro-3-cyano-6- (2-fluoro-5- { [ (5-fluoropyridin-2-yl) carbonyl]Amino } phenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-8-yl radical]Tert-butyl carbamate (40 mg, 77% yield).
The following intermediates were prepared according to a similar synthetic procedure described in example a 100:
examples A101
(R)-[2- Chlorine -3- Cyano radical -6-(2- Fluorine -5-{[(5- Methoxy pyrazine -2- Base of ) Carbonyl radical ] Amino group } Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -2-chloro-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-methoxypyrazine-2-carboxylic acid.
Examples A102
(R)-[2- Chlorine -6-(5-{[(5- Chloropyridines -2- Base of ) Carbonyl radical ] Amino group }-2- Fluorophenyl group )-3- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -2-chloro-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-chloropyridine-2-carboxylic acid.
Examples A103
(R)-[2- Chlorine -3- Cyano radical -6-(5-{[(5- Cyanopyridines -2- Base of ) Carbonyl radical ] Amino group }-2- Fluorophenyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -2-chloro-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-cyanopyridine-2-carboxylic acid.
Examples A104
(R)-{2- Chlorine -3- Cyano radical -6-[5-({[1-( Difluoromethyl group )-1H- Pyrazoles -3- Base of ] Carbonyl radical } Amino group )-2- Fluorophenyl group ]-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of } Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -2-chloro-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 1-difluoromethyl-1H-pyrazole-3-carboxylic acid.
Examples A105
(R)-[3- Cyano radical -6-(2- Fluorine -5-{[(5- Methoxy pyrazine -2- Base of ) Carbonyl radical ] Amino group } Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-methoxypyrazine-2-carboxylic acid.
Examples A106
(R)-{3- Cyano radical -6-[5-({[1-( Difluoromethyl group )-1H- Pyrazoles -3- Base of ] Carbonyl radical } Amino group )-2- Fluorophenyl group ]-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of } Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 1-difluoromethyl-1H-pyrazole-3-carboxylic acid.
Examples A107
(R)-[3- Cyano radical -6-(2- Fluorine -5-{[(5- Fluoropyridines -2- Base of ) Carbonyl radical ] Amino group } Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-fluoro-2-pyridinecarboxylic acid.
Examples A108
(R)-[3- Cyano radical -6-(5-{[(5- Cyanopyridines -2- Base of ) Carbonyl radical ] Amino group }-2- Fluorophenyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-cyanopyridine-2-carboxylic acid.
Examples A109
(R)-[6-(5-{[(5- Chloropyridines -2- Base of ) Carbonyl radical ] Amino group }-2- Fluorophenyl group )-3- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-chloropyridine-2-carboxylic acid.
Examples A110
(R)-[2,3- Dichloro (phenyl) methane -6-(5-{[(5- Cyanopyridines -2- Base of ) Carbonyl radical ] Amino group }-2- Fluorophenyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -2, 3-dichloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-cyanopyridine-2-carboxylic acid.
Examples A111
(R)-{2,3- Dichloro (phenyl) methane -6-[5-({[1-( Difluoromethyl group )-1H- Pyrazoles -3- Base of ] Carbonyl radical } Amino group )-2- Fluorophenyl group ]-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of } Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -2, 3-dichloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 1-difluoromethyl-1H-pyrazole-3-carboxylic acid.
Examples A112
(R)-[2,3- Dichloro (phenyl) methane -6-(5-{[(5- Chloropyridines -2- Base of ) Carbonyl radical ] Amino group }-2- Fluorophenyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -2, 3-dichloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-chloro-pyridine-2-carboxylic acid.
Examples A113
(R)-[2,3- Dichloro (phenyl) methane -6-(2- Fluorine -5-{[(5- Fluoropyridines -2- Base of ) Carbonyl radical ] Amino group } Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -2, 3-dichloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-fluoro-2-pyridinecarboxylic acid.
Examples A114
(R)-[2,3- Dichloro (phenyl) methane -6-(2- Fluorine -5-{[(5- Methoxy pyrazine -2- Base of ) Carbonyl radical ] Amino group } Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -2, 3-dichloro-6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-methoxypyrazine-2-carboxylic acid.
Examples A115
(R)-[6-(5-{[(5- Chloropyridines -2- Base of ) Carbonyl radical ] Amino group }-2- Fluorophenyl group )-3-( Difluoromethyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -3- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-chloro-pyridine-2-carboxylic acid.
Examples A116
(R)-[6-(5-{[(5- Cyanopyridines -2- Base of ) Carbonyl radical ] Amino group }-2- Fluorophenyl group )-3-( Difluoromethyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Carbamic acid tert-butyl esterPreparation of esters
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -3- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-cyanopyridine-2-carboxylic acid.
Examples A117
(R)-[3-( Difluoromethyl group )-6-(2- Fluorine -5-{[(5- Methoxy pyrazine -2- Base of ) Carbonyl radical ] Amino group } Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -3- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 5-methoxypyrazine-2-carboxylic acid.
Examples A118
(R)-[3-( Difluoromethyl group )-6-(2- Fluorine -5-{[(5- Fluoropyridines -2- Base of ) Carbonyl radical ] Amino group } Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Preparation of tert-butyl carbamate
From (R)-[6-(5- Amino group -2- Fluorophenyl group )-3-( Difluoromethyl group )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Carbamic acid tert-butyl ester and 5- fluorine -2- Pyridine carboxylic acid preparation.
Examples A119
(R)-{3-( Difluoromethyl group )-6-[5-({[1-( Difluoromethyl group )-1H- Pyrazoles -3- Base of ] Carbonyl radical } Amino group )-2- Fluorophenyl group ]-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of } Preparation of tert-butyl carbamate
Prepared from tert-butyl (R) - [6- (5-amino-2-fluorophenyl) -3- (difluoromethyl) -6-methyl-5, 6-dihydroimidazo [1,2-a ] pyrazin-8-yl ] carbamate and 1-difluoromethyl-1H-pyrazole-3-carboxylic acid.
A.Preparation of the Final Compounds
Examples B1
rac-6- Methyl radical -6-(3- Pyrimidines -5- Base of - Phenyl radical )-5,6- Dihydro - Imidazo [1,2-a]- Pyrazine esters -8- Preparation of amines
Tetrakis (triphenylphosphine) palladium (0) (0.027 g, 0.023 mmol) was added to stirred rac-6- (3-bromo-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] under nitrogen at room temperature]Pyrazin-8-ylamine (0.14 g, 0.46 mmol), pyrimidine-5-boronic acid (0.17 g, 1.38 mmol) and potassium carbonate (0.19 g, 1.38 mmol) were suspended in 1, 4-dioxane (4 mL) and ethanol (0.4 mL). The mixture was stirred at 150 ℃ for 30 minutes under microwave irradiation. The mixture was diluted with water and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica gel; 0/100-3/97 in 7M ammonia/MeOH/DCM). The desired fractions were collected and concentrated in vacuo to afford rac-6-methyl-6- (3-pyrimidin-5-yl-phenyl) -5, 6-dihydro-imidazo [1,2-a ] as a white solid]Pyrazin-8-ylamine (0.078 g, 56% yield).
Examples B2
rac-6-[3-(5- Methoxy radical - Pyridine compound -3- Base of )- Phenyl radical ]-6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -8- Preparation of amines
Tetrakis (triphenylphosphine) palladium (0) (0.027 g, 0.023 mmol) was added to stirred rac-6- (3-bromo-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] under nitrogen at room temperature]Pyrazin-8-ylamine (0.14 g, 0.46 mmol), 5-methoxy-3-pyridylboronic acid (0.21 g, 1.38 mmol) and potassium carbonate (0.19 g, 1.38 mmol) were suspended in 1, 4-dioxane (4 mL) and ethanol (0.4 mL). The mixture was stirred at 150 ℃ for 30 minutes under microwave irradiation. The mixture was diluted with water and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtered and the solvent evaporated in vacuo. The crude product is chromatographed on flash column (silica gel; 7M ammonia/MeOH;)0/100-3/97) in DCM solution. The desired fractions were collected and concentrated in vacuo to yield rac 6- [3- (5-methoxy-pyridin-3-yl) -phenyl ] as a white solid]-6-methyl-5, 6-dihydro-imidazo [1,2-a [ ]]Pyrazin-8-ylamine (0.080 g, 52% yield).
Examples B3
rac-6-(3',5'- Dichloro (phenyl) methane - Biphenyl -3- Base of )-6- Methyl radical -5,6- Dihydro - Imidazo -[1,2-a] Pyrazine esters -8- Preparation of amines
Tetrakis (triphenylphosphine) palladium (0) (0.029 g, 0.025 mmol) was added to stirred rac-6- (3-bromo-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] at room temperature under nitrogen]Pyrazin-8-ylamine (0.15 g, 0.5 mmol), 3, 5-dichlorophenylboronic acid (0.11 g, 0.6 mmol) and potassium carbonate (0.21 g, 1.5 mmol) in suspension in 1, 4-dioxane (4 mL) and ethanol (0.4 mL). The mixture was stirred at 60 ℃ for 18 h. The mixture was then diluted with water and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM solution 0/100-3/97 followed by EtOAc/MeOH 0/100-10/90). The desired fractions were collected and concentrated in vacuo to afford rac-6- (3',5' -dichloro-biphenyl-3-yl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a white solid]Pyrazin-8-ylamine (0.114 g, 61% yield).
Examples B4
(R)-6- Methyl radical -6-(3- Pyrimidines -5- Base of - Phenyl radical )-5,6- Dihydro - Imidazo [1,2-a]- Pyrazine esters -8- Preparation of amines
Tetrakis (triphenylphosphine) palladium (0) (0.038 g, 0.033 mmol) was added to stirred (R) -6- (3-bromo-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] under nitrogen at room temperature]Pyrazin-8-ylamine (0.20 g, 0.66 mmol), pyrimidine-5-boronic acid (0.24 g, 1.97 mmol) and potassium carbonate (0.27 g, 1.97 mmol) were suspended in 1, 4-dioxane (4 mL) and ethanol (0.4 mL). The mixture was stirred at 150 ℃ for 30 minutes under microwave irradiation. The mixture was diluted with water and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica gel; 0/100-3/97 in 7M ammonia/MeOH/DCM). The desired fractions were collected and concentrated in vacuo to give (R) -6-methyl-6- (3-pyrimidin-5-yl-phenyl) -5, 6-dihydro-imidazo [1,2-a ] as a white solid]Pyrazin-8-ylamine (0.112 g, 56% yield).
Examples B5
(R)-6-(3',5'- Dichloro (phenyl) methane - Biphenyl -3- Base of )-6- Methyl radical -5,6- Dihydro - Imidazo -[1,2-a] Pyrazine esters -8- Preparation of amines
Tetrakis (triphenylphosphine) palladium (0) (0.028 g, 0.025 mmol) was added to stirred (R) -6- (3-bromo-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] at room temperature under nitrogen]Pyrazin-8-ylamine (0.15 g, 0.5 mmol), 3, 5-dichlorophenylboronic acid (0.11 g, 0.6 mmol) and potassium carbonate (0.20 g, 1.5 mmol) were suspended in 1, 4-dioxane (4 mL) and ethanol (0.4 mL). The mixture was stirred at 60 ℃ for 18 h. The mixture was diluted with water and extracted with DCM. Is separated byOrganic layer, drying (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM solution 0/100-3/97 followed by EtOAc/MeOH 0/100-10/90). The desired fractions were collected and concentrated in vacuo to give (R) -6- (3',5' -dichloro-biphenyl-3-yl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a white solid]Pyrazin-8-ylamine (0.92 g, 50% yield).
Examples B6
rac-5- Chlorine - Pyridine compound -2- Carboxylic acids [3-(8- Amino group -6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -6- Base of )- Phenyl radical ]- Preparation of amides
5-chloro-2-pyridinecarboxylic acid (108 mg, 0.68 mmol) was added to a solution of 4- (4, 6-dimethoxy-1, 3, 5-triazin-2-yl) -4-methylmorpholinium chloride (206 mg, 0.75 mmol) in MeOH (3 mL). The mixture was stirred at room temperature for 5 minutes. Cooling the mixture to 0 ℃ and adding rac-6- (3-amino-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ]]Pyrazin-8-ylamine (150 mg, 0.62 mmol) in MeOH (3 mL). The mixture was allowed to warm to room temperature and stirred for 3 hours. The mixture was saturated with Na2CO3And water treated and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and concentration in vacuo. The crude product is purified by flash column chromatography (silica; 7N NH)30/100-2/98 in MeOH/DCM). The desired fractions were collected and the solvent was evaporated in vacuo to yield rac-5-chloro-pyridine-2-carboxylic acid [3- (8-amino-6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a white solid]Pyrazin-6-yl) -phenyl]Amide (0.145 g, 61% yield).
Examples B7
(R)-6-(2- Fluorine -5- Pyrimidines -5- Base of - Phenyl radical )-6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -8- Preparation of amines
Tetrakis (triphenylphosphine) palladium (0) (0.009 g, 0.0077 mmol) was added to stirred (R) -6- (5-bromo-2-fluoro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] at room temperature under nitrogen]Pyrazin-8-ylamine (0.05 g, 0.15 mmol), pyrimidine-5-boronic acid (0.06 g, 0.46 mmol) and potassium carbonate (0.06 g, 0.46 mmol) were suspended in 1, 4-dioxane (2 mL) and ethanol (0.2 mL). The mixture was stirred at 150 ℃ for 30 minutes under microwave irradiation. The mixture was diluted with water and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica gel; 0/100-3/97 in 7M ammonia/MeOH/DCM). The desired fractions were collected and concentrated in vacuo. The residue was triturated with ether, filtered and dried in vacuo to give (R) -6- (2-fluoro-5-pyrimidin-5-yl-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a white solid]-pyrazin-8-ylamine (0.015 g, 30% yield).
Examples B8
(R)-5- Chlorine - Pyridine compound -2- Carboxylic acids [3-(8- Amino group -6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -6- Base of )-4- Fluorine - Phenyl radical ]- Preparation of amides
5-chloro-2-pyridinecarboxylic acid (0.07 mg, 0.45 mmol) was added to a solution of 4- (4, 6-dimethoxy-1, 3, 5-triazin-2-yl) -4-methylmorpholinium chloride (134 mg, 0.49 mmol) in MeOH (3 mL). The mixture was stirred at room temperature for 5 minutes. Cooling the mixture to 0 ℃, adding (R) -6- (5-amino-2-fluoro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ℃]Pyrazin-8-ylamine (105 mg, 0.4 mmol) in MeOH (3 mL). The mixture was allowed to warm to room temperature and stirred for 3 hours. The mixture was saturated with Na2CO3And water treated and extracted with DCM. The organic layer was separated and dried (Na)2SO4) Filtration and concentration in vacuo. The crude product is purified by flash column chromatography (silica; 7N NH)30/100-2/98 in MeOH/DCM). The desired fractions were collected and the solvent was evaporated in vacuo. The residue was triturated with ether, filtered and dried in vacuo to give (R) -5-chloro-pyridine-2-carboxylic acid [3- (8-amino-6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a white solid]Pyrazin-6-yl) -4-fluoro-phenyl]Amide (0.068 g, 42% yield).
Examples B9
(R)-5- Methoxy radical - Pyrazine esters -2- Carboxylic acids [3-(8- Amino group -6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -6- Base of )-4- Fluorine - Phenyl radical ]- Preparation of amides
5-methoxy-pyrazine-2-carboxylic acid (0.105 mg, 0.68 mmol) was added to a solution of 4- (4, 6-dimethoxy-1, 3, 5-triazin-2-yl) -4-methylmorpholinium chloride (205 mg, 0.74 mmol) in MeOH (3 mL). The mixture was stirred at room temperature for 5 minutes. Cooling the mixture to 0 ℃, adding (R) -6- (5-amino-2-fluoro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ℃]Pyrazin-8-ylamine (105 mg, 0.4 mmol) in MeOH (3 mL). Mixing the aboveThe mixture was warmed to room temperature and stirred for 18 hours. The mixture was saturated with Na2CO3And water treated and extracted with DCM. The organic layer was separated and dried (Na)2SO4) Filtration and concentration in vacuo. The crude product is purified by flash column chromatography (silica; 7N NH)30/100-3/97 in MeOH/DCM). The desired fractions were collected and the solvent was evaporated in vacuo. The residue was triturated with DIPE, filtered and dried in vacuo to give (R) -5-methoxy-pyrazine-2-carboxylic acid [3- (8-amino-6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a yellow solid]Pyrazin-6-yl) -4-fluoro-phenyl]Amide (0.100 g, 41% yield).
Examples B10
(R)-5- Fluorine - Pyridine compound -2- Carboxylic acids [3-(8- Amino group -6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -6- Base of )-4- Fluorine - Phenyl radical ]- Preparation of amides
5-fluoro-2-pyridinecarboxylic acid (0.10 mg, 0.68 mmol) was added to a solution of 4- (4, 6-dimethoxy-1, 3, 5-triazin-2-yl) -4-methylmorpholinium chloride (205 mg, 0.74 mmol) in MeOH (3 mL). The mixture was stirred at room temperature for 5 minutes. Cooling the mixture to 0 ℃, adding (R) -6- (5-amino-2-fluoro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ℃]Pyrazin-8-ylamine (160 mg, 0.62 mmol) in MeOH (3 mL). The mixture was allowed to warm to room temperature and stirred for 18 hours. The mixture was saturated with Na2CO3And water treated and extracted with DCM. The organic layer was separated and dried (Na)2SO4) Filtration and concentration in vacuo. The crude product is purified by flash column chromatography (silica; 7N NH)30/100-3/97 in MeOH/DCM). The desired fractions were collected and the solvent was evaporated in vacuo. The residue is triturated with ether, filtered and dried in vacuo to give a solution(R) -5-fluoro-pyridine-2-carboxylic acid [3- (8-amino-6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a white solid]Pyrazin-6-yl) -4-fluoro-phenyl]Amide (0.088 g, 37% yield).
Examples B11
(R)-6-(2,4- Difluoro (F) -5- Pyrimidines -5- Base of - Phenyl radical )-6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -8- Preparation of amines
Tetrakis (triphenylphosphine) palladium (0) (0.051 g, 0.044 mmol) was added to stirred (R) -6- (5-bromo-2, 4-difluoro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] at room temperature under nitrogen]Pyrazin-8-ylamine (0.30 g, 0.88 mmol), pyrimidine-5-boronic acid (0.33 g, 2.64 mmol) and potassium carbonate (0.365 g, 2.64 mmol) were suspended in 1, 4-dioxane (4 mL) and ethanol (0.4 mL). The mixture was stirred for 30 minutes at 150 ℃ under microwave irradiation. The mixture was diluted with water and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica gel; 7M ammonia/MeOH/DCM solution 0/100 to 3/97 followed by MeOH in EtOAc 20/80). The desired fractions were collected and concentrated in vacuo. The product was triturated with ether, filtered and dried in vacuo to give (R) -6- (2, 4-difluoro-5-pyrimidin-5-yl-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a white solid]Pyrazin-8-ylamine (0.110 g, 37% yield).
Examples B12
(R)-6-[5-(5- Chlorine - Pyridine compound -3- Base of )-2,4- Difluoro (F) - Phenyl radical )-6- Methyl radical -5,6- Dihydro - Imidazo [1,2-a] Pyrazine esters -8- Preparation of amines
Tetrakis (triphenylphosphine) palladium (0) (0.034 g, 0.029 mmol) was added to stirred (R) -6- (5-bromo-2, 4-difluoro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] at room temperature under nitrogen]Pyrazin-8-ylamine (0.20 g, 0.59 mmol), 5-chloro-pyridine-3-boronic acid (0.138 g, 0.88 mmol) and potassium carbonate (0.243 g, 1.76 mmol) were in suspension in 1, 4-dioxane (6 mL) and ethanol (0.6 mL). The mixture was stirred at 80 ℃ for 24 h. The mixture was then diluted with water and extracted with DCM. The organic layer was separated and dried (MgSO)4) Filtered and the solvent evaporated in vacuo. The crude product was purified by flash column chromatography (silica gel; 0/100-3/97 in 7M ammonia/MeOH/DCM). The desired fractions were collected and concentrated in vacuo. The product was triturated with DIPE, filtered and dried in vacuo to give (R) -6- [5- (5-chloro-pyridin-3-yl) -2, 4-difluoro-phenyl) -6-methyl-5, 6-dihydro-imidazo [1,2-a ] as a white solid]Pyrazin-8-ylamine (0.125 g, 57% yield).
Examples B13
(R)-N-{3-[8- Amino group -2- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a]- Pyrazine esters -6- Base of ]-4- Fluorophenyl group }-5- Fluoropyridines -2- Preparation of carboxamides
Tetrakis (triphenylphosphine) palladium (0) (17 mg, 0.015 mmol) was added to stirred (R) - [ 2-bromo-6- (2-fluoro-5- { [ (5-fluoropyridin-2-yl) carbonyl]Amino } phenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-8-yl radical]Carbamic acid tert-butyl ester (0.08)5 g, 0.15 mmol) and zinc cyanide (0.020 g, 0.17 mmol) in DMF (1.52 mL). The mixture was stirred at 110 ℃ for 16 hours. The mixture was taken up in EtOAc and washed with water. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The residue was treated with sodium cyanide (0.015 g, 0.3 mmol), potassium iodide (0.025 g, 0.15 mmol), copper iodide (0.036 g, 0.19 mmol) and N, N' -dimethylethylenediamine (0.029 mL, 0.26 mmol) in toluene (2 mL). The reaction mixture was stirred at 110 ℃ for 5 hours. The solvent was evaporated in vacuo and the residue was diluted with water and extracted with EtOAc. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The crude product was purified by flash column chromatography (silica; 7M ammonia/MeOH/DCM 0/100-10/90). The desired fractions were collected and concentrated in vacuo to give (R) -N- {3- [ 8-amino-2-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ] as an oil]Pyrazin-6-yl]-4-fluorophenyl } -5-fluoropyridine-2-carboxamide (7 mg, 11% yield).
Examples B14
(R)-N-{3-[8- Amino group -2- Chlorine -3- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds -[1,2-a] Pyrazine esters -6- Base of ]-4- Fluorophenyl group }-5- Fluoropyridines -2- Carboxamides ( Compound (I) 59) Preparation of
Trifluoroacetic acid (2.5 mL, 32.67 mmol) was added to stirred (R) - [ 2-chloro-3-cyano-6- (2-fluoro-5- { [ (5-fluoropyridin-2-yl) carbonyl]Amino } phenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-8-yl radical]Tert-butyl carbamate (0.040 g, 0.074 mmol) in DCM (5 mL). The mixture was stirred at room temperature for 30 minutes. The solvent was evaporated in vacuo and the residue was dissolved in DCM and saturated NaHCO3And (6) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The residue was triturated with an 1/1 mixture of heptane/DIPE to give (R) -N- {3- [ 8-amino-2-chloro-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ]]Pyrazin-6-yl]-4-fluorophenyl } -5-fluoropyridine-2-carboxamide (26 mg, 80% yield).
Examples B15. (R)-N-{3-[8- Amino group -3- Cyano radical -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -6- Base of ]-4- Fluorophenyl group }-5- Fluoropyridines -2- Carboxamides ( Compound (I) 66) Preparation of
Trifluoroacetic acid (1 mL, 13.06 mmol) was added to stirred (R) - [ 3-cyano-6- (2-fluoro-5- { [ (5-fluoropyridin-2-yl) carbonyl]Amino } phenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-8-yl radical]Tert-butyl carbamate (0.056 g, 0.11 mmol) in DCM (2 mL). The mixture was stirred at room temperature for 2 hours. The solvent was evaporated in vacuo and the residue dissolved in DCM and saturated NaHCO3And (6) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The residue was triturated with an 1/1 mixture of heptane/DIPE to give (R) -N- {3- [ 8-amino-3-cyano-6-methyl-5, 6-dihydroimidazo [1,2-a ]]Pyrazin-6-yl]-4-fluorophenyl } -5-fluoropyridine-2-carboxamide (35 mg, 78% yield).
Examples B16. (R)-N-{3-[8- Amino group -2- Chlorine -3- Chlorine -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -6- Base of ]-4- Fluorophenyl group }-5- Fluoropyridines -2- Carboxamides ( Compound (I) 72) And (4) preparing.
Trifluoroacetic acid (0.34 mL, 5.01 mmol) was added to stirred (R) - [ 2-chloro-3-chloro-6- (2-fluoro-5- { [ (5-fluoropyridin-2-yl) carbonyl]Amino } phenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-8-yl radical]Tert-butyl carbamate (0.044 g, 0.080 mmol) in DCM (0.77 mL). The mixture was stirred at room temperature for 30 minutes. The solvent was evaporated in vacuo and the residue dissolved in DCM and saturated NaHCO3And (6) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The residue was triturated with an 1/1 mixture of heptane/DIPE to give (R) -N- {3- [ 8-amino-2-chloro-3-chloro-6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-6-yl]-4-fluorophenyl } -5-fluoropyridine-2-carboxamide (14 mg, 39% yield).
Examples B17. (R)-N-{3-[8- Amino group -3- Difluoromethyl group -6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -6- Base of ]-4- Fluorophenyl group }-5- Fluoropyridines -2- Carboxamides ( Compound (I) 77) And (4) preparing.
Trifluoroacetic acid (0.48 mL, 6.25 mmol) was added to stirred (R) - [ 3-difluoromethyl-6- (2-fluoro-5- { [ (5-fluoropyridin-2-yl) carbonyl]Amino } phenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-8-yl radical]Tert-butyl carbamate (0.053 g, 0.09 mmol) in DCM (0.96 mL). The mixture was stirred at room temperature for 30 minutes. The solvent was evaporated in vacuo and the residue dissolved in DCM and saturated NaHCO3And (6) washing. The organic layer was separated and dried (MgSO)4) Filtration and evaporation of the solvent in vacuo. The residue was triturated with an 1/1 mixture of heptane/DIPE to give (R) -N- {3- [ 8-amino-3-difluoromethyl-6-methyl-5, 6-dihydroimidazo [1,2-a [ ]]Pyrazin-6-yl]-4-fluorophenyl } -5-fluoropyridine-2-carboxamide (36 mg, 84% yield).
Examples B18. (R)-[2- Cyano radical -6-(2- Fluorine -5-{[(5- Methoxy radical - Pyrazine esters )- Carbonyl radical ] Amino group }- Phenyl radical )-6- Methyl radical -5,6- Dihydroimidazo compounds [1,2-a] Pyrazine esters -8- Base of ] Carbamate compound 79 And (4) preparing.
5-methoxy-pyrazine-2-carboxylic acid (0.044 mg, 0.28 mmol) was added to a solution of 4- (4, 6-dimethoxy-1, 3, 5-triazin-2-yl) -4-methylmorpholinium chloride (86 mg, 0.31 mmol) in MeOH (3 mL). The mixture was stirred at room temperature for 5 minutes. Cooling the mixture to 0 ℃, adding (R) -6- (5-amino-2-fluoro-phenyl) -2-cyano-6-methyl-5, 6-dihydro-imidazo [1,2-a ℃]Pyrazin-8-ylamine (80 mg, 0.28 mmol) in MeOH (3 mL). The mixture was allowed to warm to room temperature and stirred for 18 hours. The mixture was saturated with Na2CO3And water treated and extracted with DCM. The organic layer was separated and dried (Na)2SO4) Filtration and concentration in vacuo. The crude product is purified by flash column chromatography (silica; 7N NH)3In MeOH/DCM, 0/100-3/97). The desired fractions were collected and the solvent was evaporated in vacuo. The residue was triturated with DIPE, filtered and dried in vacuo to give (R) - [ 2-cyano-6- (2-fluoro-5- { [ (5-methoxy-pyrazine) -carbonyl ] as a yellow solid]Amino } -phenyl) -6-methyl-5, 6-dihydroimidazo [1,2-a]Pyrazin-8-yl radical]Carbamate (0.013 g, 11% yield).
Compounds 1-80 in tables 1-2 list compounds prepared by analogy to each example above. When no salt form is indicated, the resulting compound is the free base. 'ex No.' refers to the example numbers according to which the compounds were synthesized. By 'co. No.' is meant the compound number.
Watch (A) 1
Watch (A) 2
C. Analysis section
LCMS
For (LC) MS-characterization of the compounds of the invention, the following method was used.
General procedure A
UPLC (ultra high performance liquid chromatography) measurements were performed using an Acquity UPLC (Waters) system comprising a sampler finisher, a compound pump with exhauster (binary pump), a 4-column incubator, a Diode Array Detector (DAD) and a column as indicated in each method. The fluid from the column is directed to the MS spectrometer. The MS detector was equipped with an electrospray ionization source. Mass spectra were obtained with a single quadrupole SQD detector by scanning from 100 to 1000 in 0.1 seconds using an interchannel delay of 0.08 seconds. The capillary needle voltage was 3.0 kV. The cone voltage is 25V for the positive ionization mode and 30V for the negative ionization mode. Nitrogen was used as the nebulizer gas. The source temperature was maintained at 140 ℃. Data were collected using MassLynx-Openlynx software.
Method of producing a composite material 1
In addition to the general procedure: reverse phase UPLC was performed at 50 ℃ using a BEH-C18 column from Waters (1.7 μm, 2.1X 50 mm) at a flow rate of 1.0 ml/min, without dispensing to the MS detector. The gradient conditions used were: 95% A (6.5 mM ammonium acetate in H)295/5 in O/acetonitrile), 5% B (acetonitrile) to 40% A, 60% B (within 3.8 minutes), to 5% A, 95% B (within 4.6 minutes), held to 5.0 minutes. The injection volume was 2. mu.l.
Method of producing a composite material 2
In addition to the general procedure: reverse phase UPLC was performed at 50 ℃ using the RRHD Eclipse Plus-C18 from Agilent (1.8 μm, 2.1X 50 mm) at a flow rate of 1.0 ml/min without dispensing to the MS detector. The gradient conditions used were: 95% A (6.5 mM ammonium acetate in H)2O/acetonitrile 95/5), 5% B (acetonitrile) to 40% a, 60% B (within 7.0 minutes) to 5% a, 95% B (within 8.6 minutes), held to 9.0 minutes. The injection volume was 2.0. mu.l.
Method of producing a composite material 3
A column of RRHD Eclipse Plus-C18 (1.8 μm, 2.1X 50 mm) from Agilent was used in place of the BEH column, and the same RP gradient was used as in method 1.
General procedure B
HPLC measurements were performed using an HP 1100 (agilent technologies) system including a compound pump with an exhaust, an autosampler, a column oven, a Diode Array Detector (DAD), and a column as indicated in the methods below. The fluid from the column was dispensed to the MS spectrometer. The MS detector (either SQD or TOF) is equipped with an electrospray ionization source. The source temperature was maintained at 140 ℃. Nitrogen was used as the nebulizer gas. Data were collected using MassLynx-Openlynx software.
B1: mass spectra were acquired on a single quadrupole SQD detector by scanning from 100 to 1000 in 0.1 seconds, using a 0.08 second interchannel delay. The capillary needle voltage was 3.0 kV.
B2: mass spectra were acquired on a time-of-flight (TOF) detector by scanning from 100 to 750 in 0.5 seconds, with a dwell time of 0.3 seconds. The capillary needle voltage was 2.5 kV for the positive ionization mode and 2.9 kV for the negative ionization mode. The cone voltage for both the positive and negative ionization modes was 20V. Leucine-enkephalin is the standard substance for lock-in mass correction.
Method of producing a composite material 4
In addition to the general procedure B1: reverse phase HPLC was performed on an Eclipse Plus-C18 column (3.5 μm, 2.1X 30 mm) from Agilent at 60 ℃ at a flow rate of 1.0 ml/min without dispensing to the MS detector. The gradient conditions used were: 95% A (6.5 mM ammonium acetate in H)2O/acetonitrile 95/5), 5% B (mixture of acetonitrile/methanol 1/1) to 100% B (over 5.0 minutes), held to 5.15 minutes, equilibrated to initial conditions at 5.30 minutes until 7.0 minutes. The injection volume was 2. mu.l. The cone voltage is 20V for the positive ionization mode and 30V for the negative ionization mode.
Method of producing a composite material 5
In addition to the general procedure B1: reverse phase HPLC was performed on an Eclipse Plus-C18 column (3.5 μm, 2.1X 30 mm) from Agilent at 60 ℃ at a flow rate of 1.0 ml/min without dispensing to the MS detector. The gradient conditions used were: 95% A (6.5 mM ammonium acetate in H)2O/acetonitrile 95/5), 5% B (acetonitrile/methanol, 1/1), held for 0.2 min, to 100% B (over 3.0 min), held for 3.15 min, equilibrated to initial conditions at 3.30 min until 5.0 min. The injection volume was 2. mu.l. ConeThe voltages were 20V and 50V for the positive ionization mode and 30V for the negative ionization mode.
General procedure C
LC measurements were performed using an Acquity UPLC (Waters) system including a multiplex pump, sample collator, column heater (set at 55 ℃), Diode Array Detector (DAD) and column as indicated in the methods below. The fluid from the column was dispensed to the MS spectrometer. The MS detector was equipped with an electrospray ionization source. The mass spectra were acquired by scanning from 100 to 1000 in 0.18 seconds, using a dwell time of 0.02 seconds. The capillary needle voltage was 3.5 kV and the source temperature was maintained at 140 ℃. The cone voltage is 10V for the positive ionization mode and 20V for the negative ionization mode. Nitrogen was used as the nebulizer gas. Data acquisition was performed using a Waters-MicromassMassLynx-Openlynx data System.
Method of producing a composite material 6
In addition to general procedure C, reverse phase UPLC (ultra performance liquid chromatography) was performed on a bridged ethylsiloxane/silica hybrid (BEH) C18 column (1.7 μm, 2.1X 50 mm; Waters Acquity) at a flow rate of 0.8 ml/min. Two mobile phases (10 mM ammonium acetate in H) were used2O/acetonitrile 95/5; a mobile phase B: acetonitrile) in a running gradient from 95% a and 5% B to 5% a and 95% B over 1.3 minutes and held for 0.7 minutes. An injection volume of 0.75 ml was used.
General procedure D
LC measurements were performed using a UPLC (ultra liquid chromatography) acquisition (waters) system comprising a compound pump with an exhauster, an autosampler, a Diode Array Detector (DAD) and a column as indicated in the following methods, maintaining the column temperature at 40 ℃. The fluid from the column was sent to the MS detector. The MS detector was equipped with an electrospray ionization source. Mass spectra were acquired on a Quattro detector (three quadrupole mass spectrometers from Waters) by scanning from 100 to 1000 in 0.2 seconds, with an internal scan delay of 0.1 seconds. The capillary needle voltage was 3 kV and the ionization source temperature was maintained at 130 ℃. The cone voltage for both the positive and negative ionization modes was 20V. Nitrogen was used as the nebulizer gas. Data acquisition was performed with MassLynx-Openlynx software (Waters).
Method of producing a composite material 7
In addition to general procedure D: reverse phase UPLC was performed on a Waters Acquity BEH (bridged ethylsiloxane/silica hybrid) phenyl-hexyl column (1.7 μm, 2.1 x 100 mm) at a flow rate of 0.343 ml/min. Two mobile phases (mobile phase A: 95% 7 mM ammonium acetate/5% acetonitrile; mobile phase B: 100% acetonitrile) were used to run a gradient condition from 84.2% A and 15.8% B (hold 0.49 min) to 10.5% A and 89.5% B in 2.18 min, for 1.94 min, and back to the initial condition in 0.73 min for 0.73 min. A2 ml injection volume was used.
Melting Point
The values are either peak or melting point ranges, and are based on experimental uncertainties, which are usually associated with the analytical method.
Mettler FP 81HT / FP90 Device for measuring the position of a moving object ( Watch (A) 3 To Chinese FP90 Marked )
For the various compounds, melting points were determined in open capillaries on a Mettler FP62 or Mettler FP81HT/FP90 apparatus. Melting points were determined with a temperature gradient of 1,3,5 or 10 ℃/min. The maximum temperature was 300 ℃. The melting point was read from a digital display.
DSC823e(identified by DSC in Table 3)
For the various compounds, melting points were determined by DSC823e (Mettler-Toledo). The melting point was determined with a temperature gradient of 30 ℃ per minute. The maximum temperature was 400 ℃.
Watch (A) 3: analytical data-RtMeaning the retention time (in minutes), [ M + H]+Means the protonated mass of the compound, method means the method used for (LC) MS;
n.d. means indeterminate
SFC/MS-method:
general procedure for SFC-MS:
from a source including delivery of carbon dioxide (CO)2) And modifier FCM-1200 Multi-Pump fluid control Module, CTC analytical automatic liquid sampler, the column from room temperature to 80 ℃ TCM-20000 thermal control module Berger Instrument analysis system, SFC determination. An Agilent 1100 UV photodiode array detector equipped with a high pressure flow cell resistant to 400 bar was used. The fluid from the column was dispensed to the MS spectrometer. The MS detector was equipped with an atmospheric pressure ionization source. The following ionization parameters of the Waters ZQ mass spectrometer are: corona: 9 mua, source temperature: 140 ℃, cone voltage: 30V, probe temperature: 450 ℃, extractor: 3V, desolvation gas (desolvation gas)400L/hr, and cone gas (cone gas) 70L/hr. Nitrogen was used as the nebulizer gas. Data acquisition was performed using a Waters-MicromassMassLynx-Openlynx data System.
Method of producing a composite material 1
In addition to general procedure a: SFC chiral resolution was performed in CHIRALCEL OJ DAICEL columns (5 μm, 4.6 x 250 mm) at a flow rate of 3.0 ml/min at 35 ℃. In isocratic mode, the mobile phase is CO230% iPrOH (containing 0.3% iPrNH)2)。
Method of producing a composite material 2
In addition to general procedure a: SFC chiral separation was performed on CHIRALPAK IC DAICEL columns (5 μm, 4.6X 250 mm) at a flow rate of 3.0 ml/min. In isocratic mode, the mobile phase is CO250% iPrOH (containing 0.3% iPrNH)2)。
General procedure B for SFC-MS:
analytical SFC systems were obtained from Berger instruments (Newark, DE, USA) and included delivery of carbon dioxide (CO)2) And a complex pump control module for modifiers (FCM-1200), a thermal control module for column heating controlling the temperature in the range of 1-150 ℃ (TCM2100) and column selection valves for 6 different columns (Valco, VICI, Houston, TX, USA). Photodiode array detectors (Agilent 1100, Waldbronn, Germany) were fitted with high pressure flow cells (up to 400 bar) and with CTC LC Mini PAL autosampler (Leap Technologies, Carrboro, NC, USA). ZQ mass spectrometers (Waters, Milford, MA, USA) with orthogonal Z-electrospray interfaces were connected to the SFC-system. And an integrated platform consisting of SFC ProNTo software and Masslynx software is used for instrument control, data collection and processing.
Method of producing a composite material 3
In addition to general procedure B: SFC chiral separations were performed on CHIRALCEL OD-H columns (4.6X 250 mm) at 30 ℃ and a flow rate of 3.0 ml/min. The mobile phase was 10-40% MeOH (containing 0.2% iPrNH) at a rate of 1.6%/min2)/CO2Then from 40 to 50% MeOH/CO at a rate of 5%2And held at 50% for 3.60 min.
Method of producing a composite material 4
The same gradient as in method 3 was used except that CHIRALPAK AS-H column (4.6X 250 mm) was used instead.
Method of producing a composite material 5
In addition to general procedure B: SFC was performed on CHIRALCEL OJ-H columns (4.6X 250 mm) at 30 ℃ at a flow rate of 3.0 ml/minChiral separation. The mobile phase was 15% EtOH (containing 0.2% iPrNH)2)/CO2And kept for 15 min.
Method of producing a composite material 6
In addition to general procedure B: SFC chiral separations were performed on CHIRALPAK AS-H columns (4.6X 250 mm) at 30 ℃ with a flow rate of 3.0 ml/min. The mobile phase was 5% MeOH (containing 0.2% iPrNH)2)/CO2Held for 16.16 min, then 10% speed from 5 to 40% MeOH/CO2And held at 50% for 3.34 min.
Watch (A) 4: analysis of SFC data-RtMeaning the retention time (in minutes), [ M + H]+Means the protonation mass of the compound, method means the method used for SFC/MS analysis of enantiomerically pure compounds;
isomer elution order: a means the first eluting isomer; b means the isomer eluting a second time;
optical rotation:
optical rotation was measured with a Perkin-Elmer 341 polarimeter with sodium lamp and reported as follows: [ alpha ] O (lambda, c g/100ml, solvent, T ℃).
[α]λT= (100α) / (lxc): wherein at a temperature T (DEG C) and a wavelength lambda (in nm),lis the path length (in dm) andcis at the sample concentration (in g/100 ml). If the wavelength of light used is 589 nm (line sodium D), the symbol D can be used instead. The rotation direction flag (+ or-) should always be given. When this equation is employed, the concentrations and solvents are provided in parentheses after rotation without exception. Rotation is reported in degrees, and no concentration units are given (assumed to be g/100 ml).
Watch (A) 5: analytical data-optical rotation values of enantiomerically pure compounds
NMR
For many compounds, chloroform-d (deuterated chloroform, CDCl) was used3) Or DMSO-d6(deuterated DMSO, dimethyl-d 6 sulfoxide) as solvent, run at 360 MHz, 400 MHz and 600 MHz respectively, recorded as standard pulse sequences on a Bruker DPX-360, Bruker DPX-400 or Bruker AV-500 spectrometer1H NMR spectrum. Chemical shifts () are reported in parts per million (ppm) relative to Tetramethylsilane (TMS), which is used as an internal standard.
Watch (A) 6:
Pharmacological examples
The compounds provided herein are inhibitors of β -site APP-cleaving enzyme 1 (BACE 1). BACE1, an aspartic protease inhibitory effect, is believed to be involved in the treatment of Alzheimer's Disease (AD). The production and aggregation of β -amyloid peptide (a β) from β -Amyloid Parent Protein (APP) is thought to play a key role in the pathogenesis and progression of AD. A β is produced from Amyloid Parent Protein (APP) by sequential cleavage at the N-terminus and C-terminus of the A β region by β -secretase and γ -secretase, respectively.
The compounds of formula (I) are expected to have substantially their effect on BACE1 by virtue of their ability to inhibit enzymatic activity. Table 7 shows the performance of such inhibitors tested using a biochemical based Fluorescence Resonance Energy Transfer (FRET) assay and the α lisa assay in SKNBE2 cells described below, which assays are suitable for identifying such compounds, more particularly compounds according to formula (I).
Biochemical FRET-based assays
The assay is based on the fluorescence resonance energy transfer assay (FRET). The substrate for this assay is the APP-derived 13 amino acid peptide containing the 'Swedish' Lys-Met/Asn-Leu mutation of the Amyloid Parent Protein (APP) β -secretase cleavage site. The substrate also contains two fluorophores: (7-Methoxycoumarin-4-yl) acetic acid (Mca) is a fluorescence donor, with an excitation wavelength of 320nm and an emission wavelength of 405nm and 2, 4-dinitrophenyl (Dnp) is the proprietary quencher acceptor. The distance between the two fluorophores has been chosen such that upon photoexcitation, the donor fluorescent energy is significantly quenched by the acceptor through resonance energy transfer. Upon cleavage of BACE1, the fluorophore Mca is separated from the quencher group Dnp, restoring the full fluorescence effect of the donor. The increase in fluorescence is linearly related to the rate of proteolysis (Koike H et al J. biochem. 1999, 126, 235-242).
Briefly, recombinant BACE1 protein at a final concentration of 1. mu.g/ml was incubated with 10 μm substrate in incubation buffer (40mM citrate buffer pH 5.0, 0.04% PEG, 4% DMSO) in 384-well format plates for 120 min at room temperature in the presence or absence of compound. Next, the amount of proteolysis at T =0 and T =120 (excitation at 320nm and emission at 405 nm) was directly determined by fluorescence measurement. The results are expressed as RFU as the difference between T120 and T0. By the least squares sum method,% controlMinimum size(Controlmin) a best fit curve was fitted plotted against the compound concentration. Thus, an IC can be obtained50Value (inhibitory concentration causing 50% inhibition of activity).
LC = median of low control values
= low control: not reacted with enzyme
HC = median of high control values
= high control: reacting with an enzyme
% effective rate = 100- [ (sample-LC)/(HC-LC) × 100]
% control rate = (sample/HC) × 100
% control rateMinimum size= sample-LC)/(HC-LC) × 100
The following exemplified compounds were tested essentially as described above and showed the following activities:
watch (A) 7：
n.d. means indeterminate
Cellular alpha lisa assay of SKNBE2 cells
In two alpha lisa assays, A.beta.production and secretion into the culture medium of human neuroblastoma SKNBE2 cells was quantifiedGeneral assemblyAnd the level of a β 42. The assay was based on the human neuroblastoma SKNBE2 expressing the wild-type amyloid parent protein (hAPP 695). Compound IDiluted and added to these cells, incubated for 18 hours, and then assayed for A β 42 and A βGeneral assembly. Assay of Abeta by Sandwich alpha lisaGeneral assemblyAnd a β 42. Alpha lisa is biotinylated antibody AbN/25 using streptavidin-coated beads and detection of Abeta, respectivelyGeneral assemblyAnd Ab4G8 or cAb42/26 antibody of A β 42 conjugated acceptor beads. In A betaGeneral assemblyOr a β 42, in close proximity. Excitation of the donor beads causes the release of singlet oxygen molecules, which triggers a cascade of energy transfer in the acceptor beads, resulting in luminescence. Luminescence (excitation at 650nm and emission at 615 nm) was measured after 1 hour incubation.
By the least squares sum method,% controlMinimum size(Controlmin) a best fit curve was fitted plotted against the compound concentration. Thereby obtaining an IC50Value (inhibitory concentration causing 50% inhibition of activity).
LC = median of low control values
= low control: cells not preincubated with compound, biotinylated Ab in alpha lisa
HC = median of high control values
= high control: cells not pre-cultured with Compounds
% effective rate = 100- [ (sample-LC)/(HC-LC) × 100]
% control rate = (sample/HC) × 100
% control rateMinimum size= sample-LC)/(HC-LC) × 100
The following exemplified compounds were tested essentially as described above and showed the following activities:
watch (A) 8：
Demonstration of in vivo efficacy
The Α β peptide lowering agents of the present invention may be used to treat AD in mammals such as humans or alternatively to show efficacy in animal models such as, but not limited to, mice, rats or guinea pigs. The mammal may not be diagnosed with AD, or may not have a genetic predisposition for AD, but may be transgenic such that it overproduces and eventually deposits a β in the mammal, similar to that seen in AD patients.
The a β peptide reducing agent can be administered in any standard form using any standard method. For example, but not limited to, the Α β peptide lowering agent may be in the form of a liquid, tablet or capsule that is administered orally or by injection. The Α β peptide lowering agent can be administered at any dose sufficient to significantly reduce the level of Α β peptide in blood, plasma, serum, cerebrospinal fluid (CSF) or brain.
To determine whether acute administration of an Α β 42 peptide-reducing agent will reduce Α β peptide levels in vivo, non-transgenic rodents, such as mice or rats, were used. Animals treated with an a β peptide-reducing agent are examined and the levels of soluble a β 42 and total a β in the brain are quantified by standard techniques, e.g., using ELISA, as compared to those animals that are untreated or treated with vehicle. Once the time course of onset of action can be established, the treatment time varies from hours (h) to days and is adjusted according to the results of a β 42 reduction.
A typical protocol for determining a β 42 reduction is shown, but this is only one of many variations that can be used to optimize detectable a β levels. For example, the a β peptide reducing compound is formulated in 20% hydroxypropyl β cyclodextrin. The a β peptide-reducing agent was administered to fasted animals overnight as a single oral dose (p.o.) or a single subcutaneous dose (s.c.). After a certain period of time, typically 2 or 4h (as indicated in table 19), the animals were sacrificed and a β 42 levels were analyzed.
Blood samples were collected by decapitation and exsanguination in EDTA-treated collection tubes. Blood samples were centrifuged at 1900 g for 10 minutes (min) at 4 ℃, plasma recovered, and snap frozen for later analysis. The brain was removed from the skull and the rhombohedral. The cerebellum is removed and the left and right hemispheres are separated. The left hemisphere was stored at-18 ℃ to quantify test compound levels. The right hemisphere was rinsed in Phosphate Buffered Saline (PBS) buffer and immediately frozen on dry ice and stored at-80 ℃ until homogenization for biochemical testing.
Murine brains from non-transgenic animals are resuspended in 8 volumes per gram of tissue of 0.4% DEA (diethylamine)/50 mM NaCl containing protease inhibitors (Roche-11873580001 or 04693159001), e.g. 0.158 g brain, plus 1.264 ml of 0.4% DEA. In the FastPrep-24 system (MP Biomedicals), the whole sample was homogenized with the cell lysis matrix D (MPBio #6913-100) at 6m/s for 20 seconds. The homogenate was centrifuged at 221.300 x g for 50 min. The resulting high-speed supernatant was then transferred to fresh microcentrifuge tubes (eppendorf tubes). 9 supernatants were neutralized with 1 part of 0.5M Tris-HCl pH 6.8 and used for quantification of A βGeneral assemblyAnd a β 42.
Determination of A beta in soluble fractions of brain homogenates by enzyme-linked immunosorbent assayGeneral assemblyAnd the amount of a β 42. Briefly, standard solutions (dilutions of synthetic A β 1-40 and A β 1-42, Bachem) were prepared in 1.5 ml microcentrifuge tubes in Ultrafiltration, with final concentrations between 10000-0.3 pg/ml. Shi4 samples and standards with HRPO-labeled N-terminal antibodies for A β 42 detection and with antibodies for A βGeneral assemblyThe detected biotinylated middomain antibody 4G8 was co-incubated. Then 50 μ l of conjugate/sample or conjugate/standard mixture was added to the antibody coated plate (for A β 42 detection, the capture antibody selectively recognizes the C-terminus of A β 42, antibody JRF/cA β 42/26, andfor AbetaGeneral assemblyDetection, recognition of the N-terminal of A.beta.antibody JRF/rA.beta.2). Plates were incubated overnight at 4 ℃ to form antibody-amyloid complexes. After this incubation and subsequent washing step, ELISA for a β 42 quantification was performed by adding Quanta Blu fluorescent peroxidase substrate according to the manufacturer's instructions (Pierce corp., Rockford, Il). Reading after 10-15 min (excitation 320 nm/emission 420 nm).
For A β, according to the manufacturer's instructions (Pierce Corp., Rockford, Il.)General assemblyDetection, addition of streptavidin-peroxidase-conjugate, 60 min later followed by an additional washing step and addition of Quanta Blu fluorescent peroxidase substrate. Reading after 10-15 min (excitation 320 nm/emission 420 nm).
In this model, at least a 20% reduction in Α β 42 would be advantageous compared to untreated animals.
The following exemplified compounds were tested essentially as described above and showed the following activities:
watch (A) 9:
。

Claims (8)

1. A compound of formula (I)
(I)
Or a tautomer or stereoisomeric form thereof, wherein
R1And R2Independently selected from hydrogen, halo, cyano, C1-3Alkyl, and monohalo and polyhalo-C1-3An alkyl group;
R3selected from hydrogen, C1-3Alkyl, and monohalo and polyhalo-C1-3An alkyl group;
X1、X2、X3、X4independently is C (R)4) (ii) a Wherein for X2And X4Each R4Selected from hydrogen; for X1And X3，R4Selected from hydrogen and halo;
l is a bond or-N (R)5) CO-, wherein R5Is hydrogen or C1-3An alkyl group;
ar is a homoaryl or heteroaryl group;
wherein the homoaryl is phenyl or is selected from halo, cyano, C1-3Alkyl radical, C1-3Alkoxy, monohalo and polyhalo-C1-3Phenyl substituted with one, two or three substituents of alkyl;
heteroaryl is selected from pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl and oxadiazolyl, each optionally selected from halo, cyano, C1-3Alkyl radical, C1-3Alkoxy, monohalo and polyhalo-C1-3One, two or three substituents of alkyl; or an addition salt thereof.
2. The compound of claim 1, wherein
R1And R2Independently selected from hydrogen, fluoromethyl, difluoromethyl and trifluoromethyl, chloro, bromo and cyano;
R3is C1-3Alkyl or fluoromethyl, difluoromethyl and trifluoromethyl;
X1and X3Independently CH or CF; x2And X4Is CH;
l is-N (R)5) CO-, wherein R5Is hydrogen;
ar is a homoaryl or heteroaryl group;
wherein the homoaryl is selected from halo, cyano, C1-3Alkyl and C1-3Phenyl substituted with one or two substituents of alkoxy;
heteroaryl is selected from the group consisting of pyridyl, pyrimidinyl, pyridazinyl, pyrazolyl, oxazolyl and isothiazolyl, each optionally selected from the group consisting of halo, cyano, C1-3Alkyl radical, C1-3Alkoxy, fluoromethyl, difluoromethyl and trifluoromethyl; or
Addition salts thereof.
3. The compound of claim 1, wherein
R1Is hydrogen, fluoromethyl, difluoromethyl or trifluoromethyl, chloro, bromo or cyano;
R2is hydrogen, chloro, cyano, fluoromethyl, difluoromethyl or trifluoromethyl;
R3is methyl, fluoromethyl, difluoromethyl or trifluoromethyl;
X1is CF; x2、X3、X4Is CH;
l is-N (R)5) CO-, wherein R5Is hydrogen;
ar is heteroaryl;
wherein heteroaryl is selected from pyridyl, pyrazinyl and pyrazolyl, each substituted with one or two substituents selected from chloro, fluoro, cyano, methyl, methoxy, ethoxy, fluoromethyl, difluoromethyl and trifluoromethyl; or
Addition salts thereof.
4. The compound of claim 1 wherein R is3The substituted carbon atom has the R configuration.
5. A pharmaceutical composition comprising a therapeutically effective amount of a compound as defined in any one of claims 1 to 4 and a pharmaceutically acceptable carrier.
6. A process for the preparation of a pharmaceutical composition as defined in claim 5, characterized in that a pharmaceutically acceptable carrier is intimately mixed with a therapeutically effective amount of a compound as defined in any one of claims 1 to 4.
7. Use of a compound as defined in any one of claims 1 to 4 or a pharmaceutical composition as defined in claim 5 in the manufacture of a medicament for the treatment of a disorder selected from: alzheimer's disease, mild cognitive impairment, senility, dementia, cerebral amyloid angiopathy and down's syndrome.
8. The use according to claim 7, wherein the disorder is selected from the group consisting of dementia with lewy bodies, dementia with multiple cerebral infarcts, dementia associated with stroke, dementia associated with parkinson's disease and dementia associated with amyloid-beta.
HK14102157.9A 2010-12-22 2011-12-21 5,6-dihydro-imidazo[1,2-a] pyrazin-8-ylamine derivatives useful as inhibitors of beta-secretase (bace) HK1189221B (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP10196568.9 2010-12-22
EP10196568 2010-12-22
PCT/EP2011/073522 WO2012085038A1 (en) 2010-12-22 2011-12-21 5,6-DIHYDRO-IMIDAZO[1,2-a]PYRAZIN-8-YLAMINE DERIVATIVES USEFUL AS INHIBITORS OF BETA-SECRETASE (BACE)

Publications (2)

Publication Number Publication Date
HK1189221A1 HK1189221A1 (en) 2014-05-30
HK1189221B true HK1189221B (en) 2016-10-28

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