CN101511787B - Substituted N-phenylmethyl -5-oxo-proline-2-amides as P2X7-receptor antagonists and their methods of use - Google Patents
Substituted N-phenylmethyl -5-oxo-proline-2-amides as P2X7-receptor antagonists and their methods of use Download PDFInfo
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- 0 CC(*)(C1(C)*)C(*)(C(NCc2c(*)c(*)c(*)c(*)c2*)=O)N(*)C1=O Chemical compound CC(*)(C1(C)*)C(*)(C(NCc2c(*)c(*)c(*)c(*)c2*)=O)N(*)C1=O 0.000 description 4
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Abstract
The present invention relates to novel oxo-prolinamide derivatives of formula (I) which modulate P2X7 receptor function and are capable of antagonizing the effects of ATP at the P2X7 receptor and the use of such compounds or pharmaceutical compositions thereof in the treatment of disorders mediated by the P2X7 receptor, for example pain, inflammation and neurodegeneration.
Description
The present invention relates to heterocyclic amide derivative, it regulates the P2X7 function of receptors, and can antagonism ATP to the effect (P2X7 receptor antagonist) of P2X7 acceptor; The preparation method who relates to them; Relate to the pharmaceutical composition that contains them; With relate to the purposes of these compounds in treatment.
The P2X7 acceptor is a ligand-gated ion channel, it is expressed in hematopoietic lineage cell, for example scavenger cell, microglia, mastocyte and lymphocyte (T and B) (referring to, for example, Collo waits people Neuropharmacology, Vol.36, pp1277-1283 (1997)), and this receptor is by extracellular Nucleotide, and especially Triphosaden (ATP) activates.The activation that has shown the P2X7 acceptor is relevant with following behavior: giant cells formation, threshing, cytolytic necrocytosis, CD62L come off, regulate cell proliferation and pro-inflammatory cytokine discharges, as interleukin-11 β (IL-1 β) (as Ferrari, Deng the people, J.Immunol., Vol.176, pp3877-3883 (2006)) and tumor necrosis factor alpha (TNF α) (, wait people Journal ofNeurochemistry as Hide, Vol.75, pp965-972 (2000)).The P2X7 acceptor also is positioned at antigen presenting cell, keratinocyte, parotid gland cell, liver cell, red corpuscle, erythroleukemia cell, monocyte, inoblast, medullary cell, neurone and mesangial cell.And the P2X7 acceptor is expressed in maincenter and peripheral nervous system by presynaptic ending, and shows the release of regulating the L-glutamic acid in the neurogliocyte (Vol.50, page 92 (2000) for Anderson, people Drug.Dev.Res. such as C.).
The P2X7 receptor mapping is in immune key cells, with and have and make the ability that discharges important inflammatory mediator by these cells show the latent effect of P2X7 receptor antagonist in the large-scale disease of treatment, described disease comprises pain and neurodegenerative disease.In the nearest preclinical body research directly shown the P2X7 acceptor relevant with inflammation and neuropathic pain (Dell ' people such as Antonio, Neurosci.Lett., Vol.327, pp87-90 (2002), .Chessell, IP., wait the people, Pain, Vol.114, pp386-396 (2005), people such as Honore, J.Pharmacol.Exp.Ther., Vol.319, and in vitro study shows that the P2X7 acceptor regulates death (Skaper, the S.D. of the microgliacyte mediation of cortical neuron p1376-1385 (2006)),, Deng the people, Glia, Vol.54, p234-242 (2006)).And, in the transgene mouse model of degenerative brain disorder, observed the incremental adjustments (Parvathenani, people J.Biol.Chem.s such as L., Vol.278 (15), pp13309-13317 (2003)) of P2X7 acceptor for the amyloid-beta spot.
The invention provides regulate the P2X7 function of receptors and can antagonism ATP to the compound (P2X7 receptor antagonist) of the effect of P2X7 acceptor.In first aspect, provide formula (I) compound or its pharmacy acceptable salt:
(I)
Wherein:
R
1Expression C
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl, C
3-6Methyl cycloalkyl-or pyridylmethyl-, any of these group is optional to be replaced by 1,2 or 3 halogen atom; Or unsubstituted phenyl or benzyl;
R
2And R
3Represent hydrogen, halogen, C independently
1-6Alkyl, arylmethyl-, C
2-6Alkenyl, C
2-6Alkynyl or C
3-6Methyl cycloalkyl-; And any described C
1-6Alkyl, arylmethyl-, C
2-6Alkenyl, C
2-6Alkynyl or C
3-6Methyl cycloalkyl-optional by 1,2 or 3 halogen atom replacement;
R
4, R
5And R
6Represent hydrogen, fluorine or methyl independently; And
R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, C independently
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl or phenyl, and any described C
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl or phenyl are optional to be replaced by 1,2 or 3 halogen atom; Or R
10And R
11Form phenyl ring with the carbon atom that links to each other with them, it is optional by 1,2 or 3 halogen atom replacement;
Condition is to work as R
7And R
11When all being selected from hydrogen or fluorine, R
8, R
9And R
10In at least one is a halogen atom, or R
8, R
9And R
10Be selected from hydrogen and CF
3, and R
8, R
9And R
10In one but at the most one be CF
3
In one embodiment, provide formula (I) compound or its pharmacy acceptable salt:
(I)
Wherein:
R
1Expression C
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl or C
3-6Methyl cycloalkyl, any of these group can be chosen wantonly by 1,2 or 3 halogen atom and replace; Or unsubstituted phenyl or benzyl;
R
2And R
3Represent hydrogen, halogen, C independently
1-6Alkyl, arylmethyl, C
2-6Alkenyl, C
2-6Alkynyl or C
3-6Methyl cycloalkyl; Any described C
1-6Alkyl, arylmethyl, C
2-6Alkenyl, C
2-6Alkynyl or C
3-6Methyl cycloalkyl can be chosen wantonly by 1,2 or 3 halogen atom and replace;
R
4, R
5And R
6Represent hydrogen or fluorine independently; And
R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, C independently
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl or phenyl; And any described C
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl or phenyl can be chosen wantonly by 1,2 or 3 halogen atom and replace;
Condition is to work as R
7And R
11When representing hydrogen or fluorine independently, R
8, R
9And R
10In at least one is a halogen atom.
In one embodiment, provide formula (I) compound or its pharmacy acceptable salt:
(I)
Wherein:
R
1Expression C
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl, C
3-6Methyl cycloalkyl-or pyridylmethyl-, any of these group is optional to be replaced by 1,2 or 3 halogen atom; Or R
1Represent unsubstituted phenyl or benzyl;
R
2And R
3Represent hydrogen, halogen, C independently
1-6Alkyl, arylmethyl-, C
2-6Alkenyl, C
2-6Alkynyl or C
3-6Methyl cycloalkyl-; And any described C
1-6Alkyl, arylmethyl-, C
2-6Alkenyl, C
2-6Alkynyl or C
3-6Methyl cycloalkyl-optional by 1,2 or 3 halogen atom replacement;
R
4, R
5And R
6Represent hydrogen, fluorine or methyl independently; And
R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, C independently
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl or phenyl, and any described C
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl or phenyl are optional to be replaced by 1,2 or 3 halogen atom; Or R
10And R
11Form phenyl ring with the carbon atom that links to each other with them, it is optional by 1,2 or 3 halogen atom replacement;
Condition is to work as R
7And R
11When all being selected from hydrogen or fluorine, R
8, R
9And R
10In at least one is a halogen atom, or R
8, R
9And R
10In at the most one be CF
3Group.
Term used herein " alkyl " (when as group or group a part of) is meant the hydrocarbon chain that contains the some amount carbon atom of straight or branched.For example, C
1-6Alkyl refers at least 1 and the hydrocarbon chain of 6 carbon atoms at the most of containing of straight or branched.The example of alkyl includes, but are not limited to: methyl (Me), ethyl (Et), n-propyl, sec.-propyl, n-hexyl and isohexyl.
Term used herein " alkenyl " is meant the hydrocarbon chain that contains the some amount carbon atom of straight or branched, and wherein at least one C-C is two keys.Non-limiting examples of alkenyls includes, but are not limited to: vinyl, propenyl, n-butene base, isobutenyl, positive pentenyl and isopentene group.
Term used herein " alkynyl " is meant the hydrocarbon chain that contains the some amount carbon atom of straight or branched, and wherein at least one C-C is a triple bond.The example of alkynyl includes, but are not limited to ethynyl, proyl, butynyl, isoamyl alkynyl, positive pentynyl, dissident's alkynyl and positive hexin base.
Term ' cycloalkyl ' (except that other explanation is arranged) is meant closed 3-6 unit non-aromatic ring, for example cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
Term used herein ' aryl ' is meant C
6-10Monocycle or dicyclic hydrocarbon ring, wherein at least one ring is aromatic nucleus.These examples of groups comprise phenyl and naphthyl.
Term ' halogen used herein ' (except that other explanation is arranged) is meant the group that is selected from fluorine, chlorine, bromine or iodine.
In some embodiments of the present invention, R
1Expression C
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl or pyridylmethyl-, any of these group is optional to be replaced by 1,2 or 3 halogen atom; Or R
1Represent unsubstituted phenyl or benzyl.In one embodiment, R
1Represent unsubstituted C
1-6Alkyl, C
2-6Alkenyl, C
2-6Alkynyl, C
3-6Cycloalkyl, pyridylmethyl-, phenyl or benzyl.In another embodiment, R
1Represent unsubstituted C
1-4Alkyl, C
3-5Cycloalkyl, pyridylmethyl-, phenyl or benzyl.In another embodiment, R
1Expression methyl or ethyl.
In some embodiments of the present invention, R
2And R
3Represent hydrogen, halogen, C independently
1-6Alkyl, benzyl, C
2-6Alkenyl, C
2-6Alkynyl or C
3-6Methyl cycloalkyl-; And any described C
1-6Alkyl, benzyl, C
2-6Alkenyl, C
2-6Alkynyl or C
3-6Methyl cycloalkyl-can choose wantonly by 1,2 or 3 halogen atom replacement.
In one embodiment, R
2And R
3Represent hydrogen or halogen independently; Unsubstituted C
1-6Alkyl, benzyl, C
2-6Alkenyl, C
2-6Alkynyl or C
3-6Methyl cycloalkyl-.
In another embodiment, R
2And R
3Represent hydrogen, fluorine or methyl independently.In another embodiment, R
2And R
3All represent hydrogen.
In one embodiment of the invention, R
4And R
5Represent hydrogen or methyl independently.In another embodiment, R
6Expression hydrogen or methyl.In another embodiment, R
4, R
5And R
6All represent hydrogen.
In another embodiment of the invention, R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, trifluoromethyl or unsubstituted C independently
1-6Alkyl; Or R
10And R
11Form unsubstituted phenyl ring with the carbon atom that links to each other with them.In another embodiment, R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, methyl or trifluoromethyl independently; Or R
10And R
11Form unsubstituted phenyl ring with the carbon atom that links to each other with them.In another embodiment, R
7, R
8, R
9, R
10And R
11Represent hydrogen, chlorine, fluorine, bromine, methyl or trifluoromethyl independently.
In one embodiment of the invention, provide formula (I) compound or its pharmacy acceptable salt, wherein:
R
1Represent unsubstituted C
1-4Alkyl, C
2-4Alkenyl, C
3-5Cycloalkyl, pyridylmethyl-, phenyl or benzyl;
R
2And R
3All represent hydrogen;
R
4, R
5And R
6Represent hydrogen or methyl independently; And
R
7, R
8, R
9, R
10And R
11Represent hydrogen, chlorine, fluorine, bromine, methyl or trifluoromethyl independently;
Condition is to work as R
7And R
11When all being selected from hydrogen or fluorine, R
8, R
9And R
10In at least one is a halogen atom, or R
8, R
9And R
10Be selected from hydrogen and CF
3, and R
8, R
9And R
10In one but at the most one be CF
3
Particular compound of the present invention comprises compound or its pharmacy acceptable salt of following embodiment 1-136.
The antagonist of P2X7 can be used for prevention, treats or alleviates multiple pain status (as neuropathic pain, chronic inflammatory pain and visceral pain), inflammation and neurodegeneration, especially degenerative brain disorder.The P2X7 antagonist also can be used for forming the therapeutical agent of handling rheumatoid arthritis and inflammatory bowel.
The P2X7 function of receptors can be regulated and can antagonism ATP the competitive antagonist of function of receptors, inverse agonist, negative allosteric modulators (negative allosteric modulator) or indirect regulation agent can be the The compounds of this invention (P2X7 receptor antagonist) of the effect of P2X7 acceptor.
Some formulas (I) compound can form its acid salt in some cases.It is understandable in medicine formula (I) compound being used as salt, and salt should be pharmaceutically acceptable in these cases.Pharmacy acceptable salt comprises Berge, Bighley and Monkhouse, J.Pharm.Sci., 1977,66, those that describe among the 1-19.Formula (I) compound of alkalescence can form salt with pharmaceutically acceptable acid, and described acid comprises inorganic and organic acid.These acid comprise acetate, Phenylsulfonic acid, phenylformic acid, camphorsulfonic acid, citric acid, ethyl sulfonic acid, fumaric acid, glyconic acid, L-glutamic acid, Hydrogen bromide, hydrochloric acid, isethionic acid, lactic acid, toxilic acid, oxysuccinic acid, amygdalic acid, methylsulfonic acid, glactaric acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succsinic acid, sulfuric acid, tartrate, tosic acid etc.
The example of pharmacy acceptable salt comprises by toxilic acid, fumaric acid, phenylformic acid, xitix, pamoic acid, succsinic acid, hydrochloric acid, sulfuric acid, two dimethylene Whitfield's ointments (bismethylenesalicylic), methylsulfonic acid, ethionic acid, propionic acid, tartrate, Whitfield's ointment, citric acid, glyconic acid, aspartic acid, stearic acid, palmitinic acid, methylene-succinic acid, hydroxyethanoic acid, para-amino benzoic acid, L-glutamic acid, Phenylsulfonic acid, the cyclohexyl thionamic acid, the salt that phosphoric acid and nitric acid form.
Formula (I) compound can be prepared into crystal or amorphous form, and if for crystal may optionally be solvate, as hydrate.The present invention comprises stoichiometric solvate (as hydrate) in its scope, and the compound that contains variable quantity solvent (as water).
Formula (I) compound can exist with stereoisomeric forms in any ratio (as diastereomer and enantiomer), and the present invention includes in these stereoisomeric forms in any ratio each, and comprises that its mixture comprises racemic modification.Different stereoisomeric forms in any ratio can be isolated a kind of by ordinary method from other form, maybe can obtain any specified isomer by stereospecificity or asymmetric synthesis.The stereochemistry of end product is formed among the embodiment that determines by chirality HPLC (more specifically by described method (A) in an embodiment, (B), (C) or (D)) therein, specified corresponding stereospecificity title and structure in end product, the enantiomer of wherein said product is excessive in 70%.The appointment of absolute stereo chemistry is based on the known chirality of parent material.The composition of end product passes through not identify the stereochemistry of end product among the embodiment of chirality HPLC sign therein.Yet, the chirality of main component of expection product mixtures will show the chirality of parent material, and enantiomer excessive will depend on used synthetic method and may be similar to the enantiomer that similar embodiment (if this embodiment exist enantiomer excessive) surveyed excessive.Therefore, expect that a kind of compound of chirality form can use suitable parent material to be prepared as another kind of chirality form.Perhaps, if use the racemize parent material, expection will obtain racemic product, and single enantiomer can use ordinary method to separate.The present invention also comprises any tautomeric form and composition thereof.
The present invention also comprises isotope-labeled compound, its with formula (I) and following described in those compounds identical, except one or more atoms are substituted by nucleidic mass or total mass number and nature common nucleidic mass or the different atom of total mass number.Can incorporate the isotropic substance that isotopic example in the The compounds of this invention comprises hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, iodine and chlorine into, as 3H, 11C, 14C, 18F, 123I and 125I.
The pharmacy acceptable salt that contains other isotopic The compounds of this invention of above-mentioned isotropic substance and/or other atom and described compound within the scope of the invention.Isotope-labeled The compounds of this invention (those compounds of for example incorporating radio isotope such as 3H, 14C into) can be used for medicine and/or the substrate tissue distribution is measured.Tritium is for promptly
3H and carbon-14 are promptly
14The C isotropic substance is because easy to prepare and have detectability, especially preferred.
11C and
18The F isotropic substance especially is suitable for PET (positron emission tomography),
125The I isotropic substance especially is suitable for SPECT (single photon emission computed tomoscan art), and PET and SPECT all can be used for the brain imaging.In addition, for example deuterium is promptly with heavier isotropic substance
2H replaces can obtain the treatment advantage that some brings owing to higher metabolic stability, and for example the transformation period prolongs or the required dosage reduction in the body, so this may be preferred in some cases.Isotope-labeled formula (I) compound and following The compounds of this invention usually can be by disclosed steps among following schema and/or the embodiment, and the reagent that substitutes the plain mark of No Parity with the isotope-labeled reagent that can conveniently obtain is prepared.
Perhaps, formula (I) compound can the prodrug administration." prodrug " of formula used herein (I) compound is the functional derivatives of compound, after patient's administration, and final in vivo release type (I) compound.As prodrug Medicine-feeding type (I) compound those skilled in the art can be carried out: (a) to change in vivo initial of compound; (b) change the time length that compound acts in vivo; (c) change compound transhipment or distribution in vivo; (d) change compound solubleness in vivo; (e) overcome the side effect of compound generation or other difficulty that runs into.The functional derivatives that generally is used to prepare prodrug comprises the modification of compound, and it is decomposed by chemistry or enzyme in vivo.These modifications are known for those skilled in the art.
The preparation of compound
(I)
Formula (I) compound (wherein variable as above defines) and salt thereof and solvate can be by following method preparations, and this constitutes another aspect of the present invention.
The method of preparation formula of the present invention (I) compound comprises:
(a) with the carboxylic acid (or its activatory derivative) of formula (2) and the amine coupling (seeing flow process 1) of formula (3), wherein R
1, R
2, R
3, R
4, R
5, R
6, R
7, R
8, R
9, R
10And R
11As above definition.Compound (2) and (3) are optional protected;
(b) amine of the isocyanide of the dicarbonyl compound of formula (4), formula (5) and formula (6) reacts (seeing flow process 2), wherein R down as 100 ℃ in suitable solvent such as methyl alcohol and in suitable temperature
1, R
2, R
3, R
4, R
5, R
7, R
8, R
9, R
10And R
11As above definition, and R
6=H or methyl.Compound (4), (5) and (6) are chosen wantonly protected.This method be described in the past in the chemical literature (as H.Tye, with M.Whittaker, Org.Biomol.Chem., 2004,2,813-815; G.C.B.Harriman WO 9900362A1);
(c) carry out deprotection for protected formula (I) compound.The example of protecting group and their removal method are found in T.W.Greene and P.G.M.Wuts ' Protective Groups in OrganicSynthesis ' (J.Wiley and Sons, 3
RdEd.1999); Or
(d) be other formula (I) compound with the change of formula (I) compound.The example of conventional method for transformation comprises epimerization, oxidation, reduction, alkylation, fragrant replacement, nucleophilic substitution, acid amides coupling and ester hydrolysis.
Flow process 1.
The coupling of the amine of the acid of formula (2) and formula (3) generally comprises the use activating reagent, carbodiimide, I-hydroxybenzotriazole (HOBT) or 1-hydroxyl-7-azepine benzotriazole (HOAt) as N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride or polymkeric substance-carrying, and suitable alkali such as alkyl amine (as diisopropylethylamine, N-ethylmorpholine, triethylamine) or the pyridine of optional use, in suitable solvent such as DMF and/or methylene dichloride, and suitable temperature as 0 ℃ to room temperature.Perhaps the coupling of (2) and (3) can be by in suitable solvent such as dimethyl formamide, under suitable temperature such as room temperature, with O-(7-azepine benzo triazol-1-yl)-N, N, N ', N '-tetramethyl-urea hexafluorophosphate and suitable alkyl amine such as diisopropylethylamine are handled.Perhaps, can use formula (2) compound as activatory derivative (as acyl chlorides, mixed acid anhydride, active ester (as O-acyl group-isourea)), and under this condition, method (a) generally include with amine and handle described activatory derivative (Ogliaruso, M.A.; Wolfe, J.F.in The Chemistry ofFunctional Groups (Ed. Patai, S.) Suppl.B:The Chemistry of acid derivate, Pt.1 (John Wiley and Sons, 1979), pp442-8; Beckwith, A.L.J.in The Chemistry ofFunctional Groups (Ed.Patai, S.) Suppl.B:The Chemistry of Amides (Ed.Zabricky, J.) (John Wiley and Sons, 1970), pp 73 ff).
Flow process 2.
The representational method of preparation formula (2) compound is shown in following flow process 3-9:
Flow process 3
R wherein
1, R
2, R
3, R
4And R
5As above definition, R
6=H or F, and P
1And P
2Represent suitable protecting group, as C
1-6Alkyl or P
1And P
2=H.
For the similar approach of the conversion shown in the following flow process 3 be described in the previous chemical literature (as G.Verardo, P.Geatti, E.Pol, and A.G.Giumanini, Can.J.Chem., 80:779-788 (2002); T.Godet waits the people, Organic Letters, (2004), 6 (19), 3281-3284).
Step (i) is usually included in the suitable solvent such as methyl alcohol, suitable temperature as 0 ℃ under, (7) are carried out initial processing with alkali such as sodium hydroxide, to room temperature, carry out reductive alkylation as 0 ℃ in suitable temperature then, described reductive alkylation generally includes with aldehydes or ketones and acid and carries out follow-up processing as acetate, adds reductive agent such as sodium borohydride then.
Step (ii) can spontaneously be carried out, separate in the reaction of (7) of wherein said example (9) from above-mentioned steps (i), but more typically, suitable temperature as 110 ℃ under, in suitable solvent such as toluene, heating compound (8) obtains compound (9).
Deprotection steps (iii) generally comprises the standard step that carboxylicesters is converted into acid, as uses suitable hydroxide salt (as sodium hydroxide), in suitable solvent such as methyl alcohol, suitable temperature as 0 ℃ to room temperature.
Flow process 4
R wherein
1, R
2, R
3, R
4And R
5As above definition, R
6=H or F, L
1Be suitable group such as halogen (as chlorine or bromine) or boric acid or boric acid ester, and P
3Represent suitable protecting group, as C
1-6Alkyl.
Be described in the previous chemical literature with the similar method of conversion shown in the following flow process 4 (as people such as T.Itoh, Tetrahedron., 59 (2003), 3527-3536; T.Simandan and M.B.Smith, Synthetic Communications, 26 (9), 1827-1838 (1996)).
Step (i) is usually included in the suitable solvent such as tetrahydrofuran (THF), suitable temperature as 0 ℃ to room temperature, (10) are handled with alkali such as sodium hydride and alkylating reagent such as alkyl halide, or step (i) can be included in the suitable solvent such as toluene, closes two palladiums (0) and Xantphos in appropriate catalyst as three (dibenzalacetones)
TMThe mixture of (9,9-dimethyl-4, two (diphenylphosphino) xanthenes of 5-), and suitable alkali such as cesium carbonate exist down, suitable temperature as 120 ℃ under, will (10) with aryl halide or aryl or ene boric acid (or ester) processing.
Deprotection (ii) generally comprises the standard method that carboxylicesters is converted into acid, as uses suitable hydroxide salt (as sodium hydroxide), in suitable solvent such as methyl alcohol, suitable temperature as 0 ℃ to room temperature; Or use suitable acid (as trifluoroacetic acid), in suitable solvent such as methylene dichloride, in suitable temperature as 0 ℃ to room temperature.
Flow process 5
R wherein
1, R
2, R
3, R
4And R
5As above definition, and R
6=H or F.
Be described in the previous chemical literature with the similar method of conversion shown in the following flow process 5 and (as S.Aoki, waited the people, Tetrahedron, 60 (2004) 7053-7059).
Step (i) generally comprises in reactor or sealed tube, in suitable solvent such as water, under suitable temperature such as 100-140 ℃, with or without carry out microwave radiation heating (12).
Flow process 6
R wherein
1, R
4And R
5As above definition, R
2Represent other group that above-mentioned dehydrogenation or halogen are outer, R
6=H or F, L
1Be suitable leaving group, as halogen (as chlorine or bromine), and P
4And P
5Represent suitable protecting group, be respectively as C
1-6Alkyl and C
1-6Alkoxy carbonyl.
Be described in the previous chemical literature with the similar method of conversion shown in the following flow process 6 (as A.Bassoli, wait the people, Eur.J.Org.Chem., 2005,2518-2525).
Step (i) generally comprises to be protected (13) by standard method; as in suitable solvent such as methylene dichloride; under suitable temperature such as room temperature, handle with alkoxy carbonyl acid anhydrides such as coke acid di-t-butyl ester and alkali such as triethylamine and catalyzer such as 4-dimethylaminopyridine.
Step (ii) generally comprises in suitable solvent such as tetrahydrofuran (THF), suitable temperature as-78 ℃ to room temperature, (14) are handled with alkali such as two (TMS) lithium amide and alkylating reagent such as alkyl halide.
Step (iii) generally comprises carries out deprotection with (15) by standard method, as works as P
5During for tert-butoxycarbonyl, in suitable solvent such as diox, under suitable temperature such as room temperature, use the salt acid treatment.
Step (iv) generally comprises the step described in the above-mentioned flow process 4.
Flow process 7
R wherein
1, R
2, R
3, R
4And R
5As above definition, and R
6=H or F.P
5, P
6And P
7Represent suitable protecting group, for example P
5Can be C
1-6Alkoxy carbonyl, and P
6And P
7Can be C
1-6Alkyl (P
6And P
7Needn't be identical).L
1Be suitable leaving group, as halogen (as chlorine or bromine).
Step (i) generally comprises in suitable solvent such as tetrahydrofuran (THF), suitable temperature as-78 ℃ to room temperature, (17) are handled with suitable alkali such as hexamethyldisilane base amine potassium (potassiumhexamethyldisilazide) and alkylating reagent such as alkyl halide.
Step (ii) generally comprises uses carboxylicesters to be converted into the standard method of acid, as in suitable solvent such as methylene dichloride, under suitable temperature such as room temperature, uses suitable acid (as trifluoroacetic acid) to handle.
Flow process 8
R wherein
1, R
2, R
3, R
4And R
5As above definition, and R
6=H or F.P
8, P
9And P
10Represent suitable protecting group, as P
8And P
9Be C
1-6Alkyl (P
8And P
9Needn't be identical) and P
10For from suitable non-annularity or cyclic ketone deutero-group.
Be described in the previous chemical literature with the similar method of conversion shown in the step (i)-(iii) of following flow process 8 (as J.Wehbe, wait the people, Tetrahedron:Asymmetry., 14 (2003), 1123-1126).
Step (i) generally comprises in suitable solvent such as toluene, suitable temperature as 110 ℃ under, with (19) with suitable ketone as (1R, 2R, 5R)-2-hydroxyl pinane-3-ketone and Lewis acid such as boron trifluoride-ether complex (boron trifluoride etherate) handle.
Step (ii) generally comprises in suitable solvent such as tetrahydrofuran (THF), suitable temperature as-30 ℃ under, with (20) with Grignard reagent such as methylmagnesium-bromide and alkali as 1,8-diazabicyclo [5.4.0] 11 carbon-7-alkene use undersaturated ester (21) to handle as ethyl crotonate then.
Step (iii) generally comprises uses imines to be converted into the standard method of amine, as in suitable solvent such as tetrahydrofuran (THF), under suitable temperature such as room temperature, uses suitable acid (as 15% aqueous citric acid solution) to handle.
Step (iv) generally comprises in suitable solvent such as toluene, in suitable temperature such as room temperature to 120 ℃ heating (23) down.
Step (v) generally comprises the step of describing in the above-mentioned flow process 4.
Flow process 9
R wherein
1, R
4, R
5And R
6As above definition, R
2And R
3Represent above-mentioned group except that halogen separately, L
1And L
2Be suitable leaving group, as halogen (as chlorine or bromine), and P
11Represent suitable protecting group, as trityl.
Step (i) generally comprises in suitable solvent such as dimethyl formamide, suitable temperature as 0 ℃ to room temperature, (25) are handled with alkali such as sodium hydride and alkylating reagent such as alkyl halide.
Step (ii) generally comprises in suitable solvent such as tetrahydrofuran (THF), suitable temperature as-78 ℃ to room temperature, (26) are handled with alkali such as diisopropylaminoethyl lithium and alkylating reagent such as alkyl halide.
Step (iii) generally comprises in suitable solvent such as tetrahydrofuran (THF), suitable temperature as-78 ℃ to room temperature, (27) are handled with alkali such as diisopropylaminoethyl lithium and alkylating reagent such as alkyl halide.
Step (iv) generally comprises the standard method of using pure deprotection.For example, work as P
11During for trityl, in suitable solvent such as methyl alcohol, and under suitable temperature such as room temperature, with (28) with suitable acid such as Amberlyst
Handle.
Step (v) generally comprises the standard method of using alcohol to be oxidized to corresponding carboxylic acid, as in the mixture of suitable solvent such as SODIUM PHOSPHATE, MONOBASIC aqueous buffer solution and acetonitrile, suitable temperature as 40 ℃ under, with alcohol (29) with oxygenant such as Textone, TEMPO (2,2,6,6-tetramethyl--piperidino oxyradical) and the combined treatment of SYNTHETIC OPTICAL WHITNER (chlorine bleach liquor).
Step (ii) or step (iii) optionally can omit with the preparation compound R respectively wherein
2=H or R
3=H.
The compound of general formula (3), (4), (5), (6), (7), (10), (12), (13), (17), (19), (21) and (25) is generally from commercially available or can use the method described in the chemical literature (or using similar approach) preparation by those skilled in the art.
Be correlated with, pharmacy acceptable salt can be by the preparation easily with the reaction of suitable acid or acid derivative.
Clinical indication
It is believed that The compounds of this invention can regulate the P2X7 function of receptors and can antagonism ATP to the effect of P2X7 acceptor, compound of the present invention can be used for treating pain, comprises acute pain, chronic pain, chronic arthralgia, musculoskeletal pain, neuropathic pain, inflammatory pain, visceral pain, pain with related to cancer, with migraine, the pain that tension headache is relevant with cluster headache, the pain relevant with functional bowel disorder, lower back and cervical pain, the pain relevant with sprain and strain, associability constant pain (sympathetically maintained pain); Myositis, the pain relevant, the pain relevant, pain, post-operative pain, cancer chemotherapy, headache, toothache and the dysmenorrhoea relevant with myocardial ischemia with rheumatic fever with influenza or other virus infection (as common cold).
The chronic arthralgia illness comprises rheumatoid arthritis, osteoarthritis, rheumatoid spondylitis, urarthritis and juvenile arthritis.
The pain relevant with functional bowel disorder comprises non-ucler dyspepsia, non-heart chest pain and irritable bowel syndrome.
Neuropathic pain syndrome comprises: diabetic neuropathy, sciatica, non-specific low back pain, trigeminal neuralgia, multiple sclerosis pain, fibromyalgia, the neuropathy that HIV-is relevant, postherpetic neuralgia, trigeminal neuralgia and because the pain that health wound, amputation, phantom limb syndrome, spinal operation, cancer, toxin or chronic inflammatory illness cause.And, the neuropathic pain illness comprises with common non-painful feels relevant pain, and described sensation is as " numb " (paresthesia is with insensitive), increase painful sensation (dynamic, static, hot or cold allodynia) behind (oxypathy), the non-noxious stimulation, the susceptibility of destructive stimulus is increased (hot, cold, mechanical hyperalgesia), removes the post-stimulatory lasting pain sensation (hyperpathia) or selectivity and feel disappearance or shortage (hypoalgesia) in the conduction path the susceptibility that contacts.
Can comprise heating, inflammation, immunological disease, unusual platelet function disease (as occlusive vascular disease), sexual dysfunction or erectile dysfunction with other illness of The compounds of this invention treatment; Be characterized as unusual bone metabolism or resorbent osteopathia; Non-steroidal anti-inflammatory drug (NSAID ' s) and Hemodynamics side effect, the cardiovascular disorder of COX-2 (COX-2) inhibitor; Neurodegenerative disease (neurodegenerative diseases) and neurodegeneration (neurodegeneration), post-traumatic neurodegeneration, tinnitus, to the dependency (dependence on a dependence-inducing agent) of dependence producing drug such as opioid (as morphine), CNS tranquilizer (as ethanol), incitantia (as Cocaine) and Nicotine; Type i diabetes complication, renal tubal dysfunction, hepatic insufficiency (as hepatitis, liver cirrhosis), gastrointestinal dysfunction (as diarrhoea), colorectal carcinoma, bladder hyperactivity hyperkinesia and urge incontinence.Dysthymia disorders and alcoholism also can be treated with The compounds of this invention.
Inflammation and comprise acute injury (as conjunctivitis), the inflammatory lung disease (as asthma, bronchitis, pulmonary emphysema, allergic rhinitis, respiratory distress syndrome, pigeon-fanciers' disease, farmer lung, chronic obstructive pulmonary disease (COPD), airway hyperreactivity) of skin disorder (as sunburn, burn, eczema, dermatitis, allergic dermatitis, psoriasis), meningitis, illness in eye such as glaucoma, the retinitis, retinopathy, uveitis and ocular tissue with the inflammatory conditions of described inflammation-related; Gastrointestinal tract disease (as aphthous ulcer, Crohn's disease, atopy gastritis, varioliform gastritis (gastritis varialoforme), ulcerative colitis, coeliac disease, Crohn disease, irritable bowel syndrome, inflammatory bowel, stomach and intestine reflux disease (gastrointestinal refluxdisease)); Organ transplantation and other have the illness of inflammatory component, as vascular disease, migraine, periarteritis nodosa, thyroiditis, aplastic anemia, Hodgkin's disease, scleroderma, myasthenia gravis, multiple sclerosis, sarcoidosis (sorcoidosis), nephrotic syndrome, bechet's syndrome (Bechet ' ssyndrome), oulitis, myocardial ischemia, heating, systemic lupus erythematosus, polymyositis, tendonitis, bursitis and Sjogren syndrome.
Immunological disease comprises autoimmune disorder, immune deficiency disorder or organ transplantation.
Be characterized as unusual bone metabolism or resorbent osteopathia and comprise osteoporosis (especially post-menopausal osteoporosis), hypercalcemia syndrome, hyperparathyroidism, the Paget osteopathia (Paget ' s bonediseases), osteolysis, has or do not have the hypercalcemia syndrome of the malignant tumour of bone transfer, rheumatoid arthritis, periodontitis, osteoarthritis, ostalgia, osteopenia, carcinemia, lithiasis, calculus forms (especially urolithiasis), solid carcinoma, gout and ankylosing spondylitis, tendonitis and bursitis.
Cardiovascular disorder comprises hypertension or myocardial ischemia; Atherosclerosis; Functional or organ venous insufficiency (functional or organic venous insufficiency); The treatment of varicose; Hemorrhoid; With with the inferior relevant shock state (as septic shock) that obviously descends of artery.
Neurodegenerative disease comprises dementia, especially degeneration dementia (comprise senile dementia, have dementia with Lewy body (dementia with Lewy bodies), degenerative brain disorder, Pick's disease, Huntington Chorea, Parkinson's disease and Creutzfeldt-Jakob disease, amyotrophic lateral sclerosis (ALS) and motor neurone disease); Vascular dementia (comprising multi-infarct dementia); And the dementia relevant with intracranial space occupying lesion; Wound; Infect and relevant illness (comprising HIV infection, meningitis and zoster); Metabolism; Toxin; Anoxic and vitamin deficiency; And the mild cognitive impairment relevant with the age, especially relevant memory impairment with the age.
Formula (I) compound also can be used for neuroprotective and the post-traumatic neurodegeneration of treatment, as apoplexy, asystole, pulmonary artery bypass, traumatic brain injury, Spinal injury etc.
Compound of the present invention also can be used for treating the malignant cell growth and/or shifts and the sarcoplast leukemia.
The complication of type 1 diabetes comprises diabetic microangiopathy, diabetic retinopathy, diabetic nephropathy, macular degeneration, glaucoma, nephrotic syndrome, aplastic anemia, uveitis, kawasaki disease and sarcoidosis.
Renal tubal dysfunction comprises ephritis, glomerulonephritis, especially mesangial proliferative glomerulonephritis and nephritic syndrome.
Unless should be appreciated that other by explanation, the treatment that relates to comprises the treatment of the symptom that has formed and prophylactic treatment.
According to a further aspect in the invention, we provide formula (I) compound or its pharmacy acceptable salt thus, and it is used for people or veterinary medicine.
According to a further aspect in the invention, we provide formula (I) compound or its pharmacy acceptable salt, and it is used for the treatment of the receptor-mediated illness by P2X7.
According to a further aspect in the invention, we provide and have treated the method for suffering from by the human or animal patient of the receptor-mediated illness of P2X7, and this method comprises formula (I) compound or its pharmacy acceptable salt that gives significant quantity to described patient.
According to a further aspect in the invention, we provide treatment to suffer from the human or animal patient's of pain, inflammation or neurodegenerative disease method, and this method comprises formula (I) compound or its pharmacy acceptable salt that gives significant quantity to described patient.
According to a further aspect in the invention, we provide treatment to suffer from the human or animal patient's of inflammatory pain, neuropathic pain or visceral pain method, and this method comprises formula (I) compound or its pharmacy acceptable salt that gives significant quantity to described patient.
According to a further aspect in the invention, the patient that we provide treatment to suffer from degenerative brain disorder, people patient's method for example, this method comprises formula (I) compound or its pharmacy acceptable salt that gives significant quantity to described patient.
According to a further aspect in the invention, we provide formula (I) compound or its pharmacy acceptable salt to be used for the treatment of by the purposes in the medicine of the illness that effect mediated of P2X7 acceptor in preparation.
According to a further aspect in the invention, we provide formula (I) compound or its pharmacy acceptable salt preparation be used for the treatment of or the medicine of prevent irritation, inflammation or neurodegenerative disease in purposes.
According to a further aspect in the invention, we provide formula (I) compound or its pharmacy acceptable salt to be used for the treatment of or to prevent purposes in the medicine of inflammatory pain, neuropathic pain or visceral pain in preparation.
In one aspect of the invention, we provide formula (I) compound or its pharmacy acceptable salt to be used for the treatment of or to prevent purposes in the medicine of degenerative brain disorder in preparation.
Treat people and other Mammals for use formula (I) compound or its pharmacy acceptable salt, usually it is mixed with pharmaceutical composition with the standard drug method.Therefore, provide the pharmaceutical composition that comprises formula (I) compound or its pharmacy acceptable salt in another aspect of this invention, it is suitable for people or veterinary drug.
For use formula (I) compound in treatment, with standard pharmaceutical procedures they are mixed with pharmaceutical composition usually.The present invention also provides pharmaceutical composition, and it comprises formula (I) compound or its pharmacy acceptable salt and optional pharmaceutically acceptable carrier.
That pharmaceutical composition of the present invention (its can by being mixed with under envrionment temperature and normal atmosphere) is suitable for usually is oral, parenteral or rectal administration, and can thus be tablet, capsule, oral liquid, powder, particle, lozenge, restructural powder (reconstitutable powders), injection or infusion solution or suspensoid or suppository.The composition of common preferred oral administration.
The tablet of oral administration and capsule can be unit dosage form, and can contain conventional vehicle, as tackiness agent, weighting agent, compressing tablet lubricant, disintegrating agent and acceptable wetting agent.Tablet can be according to known method dressing in the conventional pharmaceutical practice.
Oral liquid can be, the form of water-based or oiliness suspensoid, solution, emulsion, syrup or elixir for example, maybe can be the form of the drying products that is used for water before use or other suitable solvent reconstruct.These liquid preparations can contain conventional additives such as suspending agent, emulsifying agent, non-water-soluble matchmaker (it can comprise edible oil), sanitas, and optionally conventional flavouring agent or tinting material.
For parenteral admin, use compound of the present invention or its pharmacy acceptable salt and aseptic solvent to prepare the liquid unit dosage.Compound, according to used solvent and concentration, but suspendible or be dissolved in the solvent.In preparation solution, soluble compound is used for injection, and sterile filtration, is packed in suitable bottle or the ampoule then and sealing.Advantageously, with adjuvant, be dissolved in the solvent as local anaesthetics, sanitas and buffer reagent.In order to improve stability, being filled in the bottle and moving down in vacuum can be freezing with composition after dewatering.The parenteral suspensoid can essentially identical mode prepare, and except compound is suspendible rather than is dissolved in the solvent, and need not filter and sterilize.Compound can be sterilized by being exposed in the oxyethane, is suspended in then in the aseptic solvent.Advantageously, in composition, comprise tensio-active agent or wetting agent to promote the uniform distribution of compound.
Depend on medication, composition can contain 0.1% to 99% weight, the active substance of preferred 10 to 60% weight.
The dosage of used compound will change with ordinary method according to the similar factor with other of severity, weight in patients of disease in the above-mentioned disease of treatment.Yet as common guidance, suitable unitary dose can be 0.05 to 1000mg, and being more suitable for is 0.05 to 200mg, for example 20-40mg; And this unitary dose is administered once preferred every day, but may need administration every day more than once; And should treatment sustainable many weeks or a plurality of months.
All publications of being quoted in the specification sheets include but not limited to that patent and patent application are incorporated herein by reference in this article, represent all to incorporate this paper into as a reference particularly and respectively as every piece of publication.
Following specification sheets and embodiment illustrate the preparation of The compounds of this invention, but are not in order to limit the present invention.
Embodiment:
Universal method (a)-(d), the synthetic method with shown in the above-mentioned flow process 1-9 is used to prepare compound of the present invention, and these methods will further describe by following embodiment.
Embodiment 1N-[(2-chloro-4-fluorophenyl) methyl]-5-oxo-1-(phenyl methyl)-prolineamide (E1)
Under argon atmospher, with 5-oxo-1-(phenyl methyl)-proline(Pro) (0.176g, 0.80mmol, be prepared as follows) be dissolved in the methylene dichloride (3ml), and to wherein add I-hydroxybenzotriazole (0.119g, 0.88mmol), triethylamine (0.113ml, 0.81mmol), [(2-chloro-4-fluorophenyl) methyl] amine (0.134g, 0.84mmol) and N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.169g, 0.88mmol).Mixture at room temperature stirs and spends the night.Mixture dilutes with methylene dichloride, and successively with 2M aqueous hydrochloric acid and saturated sodium bicarbonate aqueous solution washing.Organic layer filters by phase splitter, and evaporation obtains crude product then.Thick material obtains pure N-[(2-chloro-4-fluorophenyl by automatic (mass-directed automated) HPLC purifying of mass spectrometric detection) methyl]-5-oxo-1-(phenyl methyl)-prolineamide (0.112g), be white solid.LC/MS[M+H]
+=361.2, retention time=2.55 minute.
5-oxo-1-(phenyl methyl)-proline(Pro) used in the aforesaid method is prepared as follows:
(i) (0.500g 2.37mmol) is dissolved in the methyl alcohol (10ml), and is cooled to 0 ℃ with L-glutamic acid dimethyl ester hydrochloride.Then with mixture with sodium hydroxide (0.099g 2.49mmol) handles, use then acetate (0.136ml, 2.37mmol) and phenyl aldehyde (0.361ml, 3.55mmol) processing.In 0 ℃ stir 10 minutes after, (0.088g 2.37mmol), and is warmed to room temperature with mixture, and stirs and spend the night to add sodium borohydride.Mixture is cooled to 0 ℃ once more, and (0.044g handles 1.18mmol) with other sodium borohydrides.Mixture is warmed to room temperature once more, and stirs and spend the night.Evaporation methyl alcohol obtains resistates, is dissolved in the ethyl acetate it and filtration.Then filtrate is washed with saturated sodium bicarbonate aqueous solution, filter (stirring simultaneously) by phase splitter, and evaporation obtains clarifying oily matter (0.56g).This oily matter is dissolved in the methyl alcohol, and in the pipe of sealing in microwave reactor in 120 ℃ of heating 10 minutes, then in 140 ℃ of heating 15 minutes (LC/MS shows that the composition of mixture between heating period does not obviously change).Evaporating solvent, and with resistates by quick silica gel column chromatography, come purifying with the hexane solution wash-out of the ethyl acetate of 15-20% gradient, obtain pure 5-oxo-1-(phenyl methyl)-proline methyl ester (0.212g), be to clarify oily matter.
LC/MS[M+H]
+=234, retention time=2.15 minute.
(ii) with 5-oxo-1-(phenyl methyl)-proline methyl ester (0.212g, 0.91mmol) in water-soluble (3ml) and the methyl alcohol (0.5ml), and with the 2M aqueous sodium hydroxide solution (0.682ml 1.36mmol) handles.Mixture at room temperature stirs and spends the night, and uses washed with dichloromethane then.The evaporation water layer, and resistates is with the (～5ml) processing of the ethereal solution of excessive 1M hydrogenchloride.Evaporating mixture once more, and resistates grinds with methylene dichloride.Discharge solid matter, and the methylene dichloride fraction (fractions) of evaporation merging, obtain 5-oxo-1-(phenyl methyl)-proline(Pro) (0.182g), be yellow oil, it is not further purified promptly and uses.
LC/MS[M+H]
+=220, retention time=1.72 minute.
Embodiment 2N-[(2-chloro-4-fluorophenyl) methyl]-1-(1-methylethyl)-5-oxo-prolineamide (E2)
Under argon atmospher, with 1-(1-methylethyl)-5-oxo-proline(Pro) (0.060g, 0.35mmol, be prepared as follows) be dissolved in methylene dichloride (3ml) and the dimethyl formamide (1ml), and to wherein adding I-hydroxybenzotriazole (0.052g, 0.39mmol), [(2-chloro-4-fluorophenyl) methyl] amine (0.061g, 0.39mmol) and N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.074g, 0.39mmol).Mixture at room temperature stirs and spends the night.Mixture dilutes with methylene dichloride, and successively with 2M aqueous hydrochloric acid and saturated sodium bicarbonate aqueous solution washing.Organic layer filters by phase splitter, and evaporation obtains crude product then.Thick material obtains pure N-[(2-chloro-4-fluorophenyl by the automatic HPLC purifying of mass spectrometric detection) methyl]-1-(1-methylethyl)-5-oxo-prolineamide (0.032g), be white solid.LC/MS[M+H]
+=313.1, retention time=2.26 minute.
1-(1-methylethyl)-5-oxo-proline(Pro) used in the aforesaid method is prepared as follows:
(i) (0.500g 2.37mmol) is dissolved in methyl alcohol (4ml) and the tetrahydrofuran (THF) (8ml), and (0.099g 2.49mmol) handled 10 minutes with the sodium hydroxide of pulverizing with mixture then with L-glutamic acid dimethyl ester hydrochloride.At this moment, with acetate (0.136ml, 2.37mmol) and acetone (0.261ml 3.55mmol) adds in the mixture of tetrahydrofuran solution (1ml) most together.Stir after 10 minutes, mixture is cooled to 0 ℃, and (0.088g 2.37mmol) handles with sodium borohydride sheet (pellets).Then mixture is warmed to room temperature, and stirs and spend the night.Evaporation methyl alcohol obtains resistates, is dissolved in the ethyl acetate it and filtration.Filtrate is washed with saturated sodium bicarbonate aqueous solution then, filter (stirring simultaneously) by phase splitter, and evaporation obtains clarifying oily matter (0.217g).Oily matter obtains pure N-(1-methylethyl)-glutamic acid dimethyl ester (0.200g) by quick silica gel column chromatography purifying.
(ii) N-(1-methylethyl)-glutamic acid dimethyl ester (0.200g) is dissolved in the methyl alcohol, and in sealed tube, in microwave reactor, heated 20 minutes in 140 ℃.Thin-layer chromatography shows that initial substance keeps intact, thereby evaporating solvent, and replaces with toluene.In reflux temperature heating～3 hours, evaporation obtained 1-(1-methylethyl)-5-oxo-proline methyl ester (0.152g) then, is faint yellow oily thing with mixture, and it is not further purified and just is used for next step.
(iii) with 1-(1-methylethyl)-5-oxo-proline methyl ester (0.152g, 0.82mmol) in water-soluble (3ml) and the methyl alcohol (0.5ml), and with the 2M aqueous sodium hydroxide solution (0.682ml 1.36mmol) handles.Mixture is used washed with dichloromethane then in stirring at room～4 hour.The evaporation water layer, and resistates is with the (～5ml) processing of the ethereal solution of excessive 1M hydrogenchloride.Evaporating mixture once more, and resistates grinds with methylene dichloride.Discharge solid matter, and the methylene dichloride fraction of evaporation merging, obtain 1-(1-methylethyl)-5-oxo-proline(Pro) (0.060g), be yellow oil, leave standstill crystallization.
Embodiment 3N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-5-oxo-prolineamide (E3)
1-ethyl-5-oxo-proline methyl ester (0.135g, 0.79mmol are prepared as follows) is dissolved in the methyl alcohol (4ml), and (0.592ml 1.18mmol) handles with the 2M aqueous sodium hydroxide solution.Mixture is in stirring at room～4 hour, and evaporation obtains resistates then, its use then excessive 1M hydrogenchloride ethereal solution (～5ml) handled 10 minutes.Evaporating mixture once more, and resistates is dissolved in methylene dichloride (4ml) and the dimethyl formamide (2ml), and filter and remove solid.Gained solution is transferred in the reaction tubes, add I-hydroxybenzotriazole (0.117g then, 0.87mmol), [(2-chloro-4-fluorophenyl) methyl] amine (0.138g, 0.87mmol) and N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.167g, 0.87mmol).The mixture argon purge is spent weekend in stirring at room then.Mixture dilutes with methylene dichloride then, and successively with 2M aqueous hydrochloric acid and saturated sodium bicarbonate aqueous solution washing.Organic layer filters by phase splitter, and evaporation obtains crude product then.Thick material obtains pure N-[(2-chloro-4-fluorophenyl by the automatic HPLC purifying of mass spectrometric detection) methyl]-1-ethyl-5-oxo-prolineamide (0.086g), be white solid.LC/MS[M+H]
+=299.1, retention time=2.13 minute.
Enantiomer is excessive=and 100.0%, detect by chiral chromatography B, be indicated as N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-5-oxo-L-prolineamide
Retention time=8.05 minute
1-ethyl-5-oxo-proline methyl ester used in the aforesaid method is prepared as follows:
(i) (0.500g 2.37mmol) is dissolved in methyl alcohol (4ml) and the tetrahydrofuran (THF) (8ml), and mixture is used the sodium hydroxide of pulverizing, and (0.099g 2.49mmol) handled 10 minutes with L-glutamic acid dimethyl ester hydrochloride.At this moment, with acetate (0.136ml, 2.37mmol) and acetaldehyde (0.199ml 3.55mmol) adds in the mixture of tetrahydrofuran solution (1ml) most together.Stir after 10 minutes, mixture is cooled to 0 ℃, and (0.088g 2.37mmol) handles with the sodium borohydride sheet.Then mixture is warmed to room temperature.After mixture reaches room temperature, it is diluted with ethyl acetate (30ml), and, filter (stirring simultaneously) by phase splitter, and evaporation obtains the oily resistates with the saturated sodium bicarbonate aqueous solution washing.This oily matter is dissolved in the toluene, and in the heating down 4 hours that refluxes.For guaranteeing to finish, then with mixture heated overnight under refluxing.Evaporating solvent then, and the gained resistates is by quick silica gel column chromatography, and gradient is that the hexane solution wash-out of 30-50% ethyl acetate comes purifying, obtains thick 1-ethyl-5-oxo-proline methyl ester (0.135g), be clarification oily matter, it is not further purified promptly and uses.
Embodiment 4-8
With with similar methods described in the foregoing description 3, prepare compound in the following table (table 1) by the acetaldehyde that replaces with suitable aldehyde (or ketone) using in the aforesaid method.The aldehyde of the compound shown in the preparation table 1 that is useful on and ketone prepare from the commercially available approach of having described in the chemical literature that maybe can use.
Table 1
Embodiment 9N-[(2-chloro-4-fluorophenyl) methyl]-5-oxo-1-phenyl-prolineamide (E9)
With 5-oxo-1-phenyl-proline(Pro) (0.047g, 0.23mmol, be prepared as follows) be dissolved in methylene dichloride (～2ml) and in the dimethyl formamide (1ml), and to wherein adding I-hydroxybenzotriazole (0.034g, 0.25mmol), [(2-chloro-4-fluorophenyl) methyl] amine (0.040g, 0.25mmol), N-ethylmorpholine (0.032ml, 0.25mmol) and N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.048g, 0.25mmol).With mixture in stirring at room 4.5 hours.Mixture is diluted with methylene dichloride again, and successively with 2M aqueous hydrochloric acid and saturated sodium bicarbonate aqueous solution washing.Organic layer filters by phase splitter, and evaporation obtains crude product then.Thick material obtains pure N-[(2-chloro-4-fluorophenyl by the automatic HPLC purifying of mass spectrometric detection) methyl]-5-oxo-1-phenyl-prolineamide (0.032g), be white solid.LC/MS[M+H]
+=347.1, retention time=2.51 minute.
5-oxo-1-phenyl-the proline(Pro) that uses in the aforesaid method is prepared as follows:
(i) with 5-oxo-L-proline methyl ester (0.204ml, 1.75mmol) be dissolved in the toluene (5ml), and with three (dibenzalacetones) close two palladiums (0) (0.024g, 0.03mmol), bromobenzene (0.184ml, 1.75mmol), cesium carbonate (0.795g, 2.45mmol) and Xantphos
TM(0.040g 0.07mmol) handles.The gained mixture is cooled to room temperature then in 120 ℃ of heating～18 hours.Mixture dilutes with ethyl acetate, and uses 2M aqueous hydrochloric acid, saturated sodium bicarbonate aqueous solution and salt water washing successively.Filter by phase splitter, then evaporation obtain yellow/brown oily matter (～0.200g).Thick material obtains pure 5-oxo-1-phenyl proline methyl ester (0.054g) by the automatic HPLC purifying of mass spectrometric detection, is oily matter, and it is left standstill crystallization.LC/MS[M+H]
+=220, retention time=2.03 minute.
(ii) (0.054g, 0.25mmol) (0.160ml, 0.32mmol) merging in methyl alcohol (1ml) is and in stirred overnight at room temperature with the 2M aqueous sodium hydroxide solution with 5-oxo-1-phenyl proline methyl ester.Evaporating solvent then, and resistates is dissolved in the ethyl acetate, and wash with the 2M aqueous hydrochloric acid.Separate water layer, and use the ethyl acetate washed twice, the ethyl acetate layer of He Binging phase splitter drying, and evaporation then obtains 5-oxo-1-phenyl-proline(Pro) (0.047g), is clarification oily matter.
Embodiment 10N-[(2, the 4-dichlorophenyl) methyl]-1-methyl-5-oxo-prolineamide (E10)
With 1-methyl-5-oxo-proline(Pro) (0.057g, 0.4mmol, be prepared as follows) be dissolved in the anhydrous methylene chloride (6ml), and to wherein adding I-hydroxybenzotriazole (0.060g, 0.4mmol), [(2,4-two chloro-phenyl) methyl] amine (0.055ml, 0.4mmol), diisopropylamine (0.140ml, 0.8mmol) and O-(7-azepine benzo triazol-1-yl)-N, N, N ', and N '-tetramethyl-urea hexafluorophosphate (0.152g, 0.4mmol).Mixture stirred 3 hours down in room temperature (20 ℃) argon gas, spent the night then.Mixture dilutes (25ml) with methylene dichloride again, and uses 2M aqueous hydrochloric acid (20ml), saturated sodium bicarbonate aqueous solution (20ml), 10% aqueous sodium carbonate (20ml) and salt solution (20ml) washing successively.Organic layer filters by the hydrophobic glass material, and evaporation obtains crude product then.Thick material is dissolved in the mixture of dimethyl sulfoxide (DMSO) (0.9ml) and acetonitrile (0.9ml), the automatic HPLC purifying by mass spectrometric detection obtains pure N-[(2, the 4-dichlorophenyl then) methyl]-1-methyl-5-oxo-prolineamide (0.085g), be white solid.LC/MS[M+H]
+=301, retention time=2.16 minute.
1-methyl-5-oxo-the proline(Pro) that uses in the aforesaid method is prepared as follows:
(i) (0.500g 3.1mmol) in water-soluble (1ml), and in microwave reactor, heated 30 minutes in 140 ℃ in sealed tube with N-methyl-L-L-glutamic acid.Vaporize water, and resistates then grinds with ether, obtains 1-methyl-5-oxo-proline(Pro) (0.298g) after the drying, is white solid.
N-[(2,4-dichlorophenyl) methyl]-1-methyl-5-oxo-prolineamide also can be prepared as follows:
With 1-methyl-5-oxo-proline(Pro) (36.79g, 0.257 mole, as above the preparation) be suspended in DCM (methylene dichloride) (500ml) in.Disposable adding EEDQ (2-oxyethyl group-1-ethoxy carbonyl-1,2-dihydroquinoline, 66.7g, 0.27 mole, 1.05 equivalents).Seem to obtain opaque mixture after all substances dissolving, and temperature is reduced to 10 ℃ from 21 ℃.Argon gas stirred 20 minutes down, dripped 2, the DCM solution (100ml) of 4-dichloro benzyl amine (36ml, 0.27 mole, 1.05 equivalents) through 40 minutes then.In the adition process, in dropping funnel, form white precipitate.Mixture slightly produces bubble, and uses ice/water-bath to be 15-20 ℃ to keep temperature.After adding amine was finished, dropping funnel used DCM (50ml) washing so that all precipitations are washed in the reaction mixture again.Then mixture is warmed to room temperature, and stirred about 18 hours.(200ml) is added in the mixture with saturated sodium bicarbonate aqueous solution, and stirs 5 minutes.Separate organic layer then, and with the HCl of 2N (3 * 250ml) washings.In the acid cleaning process, in organic layer, begin to form crystal, therefore use DCM (200ml) dilution again.Organic layer is by hydrophobic glass material drying, and vacuum concentration obtains the 65g pink solid then.Solid forms bulk, and therefore thick material grinds with pestle and alms bowl.Use ether (400ml) to grind then, and leach solid, and use Et again
2O (2 * 200ml) washings.Drying obtains 52.96g pale pink solid then.It merges (141.42g altogether) with 2 batches of materials with same procedure preparation, is suspended in then in ethanol (430ml) and the water (715ml), and is warmed to 65 ℃ (solution temperatures) gradually.Mixture obtains almost clear soln (deep pink), just has very thin suspended solid., remove flask from heating, and be warmed to ambient temperature overnight after 20 minutes in 65 ℃ of heating.Afterwards, from solution, be settled out the white needles thing.Mixture is cooled off 20 minutes to guarantee all solid precipitations in ice bath.From pink colour solution, leach white solid then, and with the EtOH/H of 3: 5 ratios
2O (2 * 400ml) (they cool off in ice bath) washing.(40 ℃) dry this solid obtained pure N-[(2,4-dichlorophenyl altogether in 5 days in vacuum drying oven) methyl]-1-methyl-5-oxo-prolineamide (125.37g), be clear crystal.
LC/MS[M+H]
+=301, retention time=2.34 minute.
1H NMR (CDCl
3, 500MHz) δ 2.01 (m, 1H), 2.34 (m, 1H), 2.37 (m, 1H), 2.46 (m, 1H), 2.80 (s, 3H), 3.99 (dd, 1H, J=9.1,4.2Hz), 4.49 (dd, 1H, J=14.9,5.9Hz), 4.55 (dd, 1H, J=14.8,6.1Hz), 6.56 (wide t, 1H, J=5.7Hz), 7.24 (dd, 1H, J=8.2,2.1Hz), 7.33 (d, 1H, J=8.2Hz), 7.40 (d, 1H, J=2.1Hz);
13C NMR δ 175.9,171.3,134.5,134.3,133.7,131.5,129.6,127.5,63.8,41.2,29.4,29.2,23.4.
Enantiomer is excessive=and 99.5%, detect by chiral chromatography A and to be indicated as N-[(2,4-dichlorophenyl) methyl]-1-methyl-5-oxo-L-prolineamide
Retention time=9.89 minute
[α]
D=-2.1 ° (c=1, MeOH), temperature=29.3 ℃, wavelength=589nm
Fusing point=144.0-144.8 ℃
Embodiment 11N-[(2-chloro-4-fluorophenyl) methyl]-1-methyl-5-oxo-prolineamide (E11)
With 1-methyl-5-oxo-proline(Pro) (0.050g, 0.35mmol, be prepared as follows) be dissolved in anhydrous methylene chloride (～7ml) in, and to wherein adding I-hydroxybenzotriazole (0.047g, 0.42mmol), [(2-chloro-4-fluorophenyl) methyl] amine (0.056ml, 0.42mmol), N-ethylmorpholine (0.166ml, 1.04mmol) and N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.067g, 0.42mmol).Mixture at room temperature stirs and spends the night.(0.100ml 0.8mmol) is added to mixture, and continues to stir the longer time, but HPLC shows do not have further product to form with [(2-chloro-4-fluorophenyl) methyl] amine of another equal portions (aliquot).Mixture is used 2M aqueous hydrochloric acid (5ml) and saturated sodium bicarbonate aqueous solution (5ml) washing successively.Collected organic layer and evaporation obtain crude product.Thick material obtains pure N-[(2-chloro-4-fluorophenyl by the automatic HPLC purifying of mass spectrometric detection) methyl]-1-methyl-5-oxo-prolineamide (0.015g), be white solid.LC/MS[M+H]
+=285, retention time=2.04 minute.
1-methyl-5-oxo-the proline(Pro) that uses in the aforesaid method is prepared as follows:
(i) (1g 6.99mmol) is dissolved in/is mixed in the tetrahydrofuran (THF) (10ml), and uses ice bath to be cooled to 0 ℃ with (L)-Pyrrolidonecarboxylic acid methyl ester.(0.201g, 60% suspension in oil 8.38mmol) is added to mixture with sodium hydride.After stopping to produce bubble, (0.522ml 8.38mmol), and is warmed to room temperature with mixture, stirs then 1 hour to add methyl iodide.Evaporating solvent, and add entry (1ml).Water layer dichloromethane extraction then.The evaporation methylene dichloride obtains thick 1-methyl-5-oxo-proline methyl ester (0.308g), and it is not further purified and just is used for next step.
(ii) with 1-methyl-5-oxo-proline methyl ester (0.308g, 1.96mmol) be dissolved in methyl alcohol (～10ml) in, and to wherein adding sodium hydroxide (0.157g, the (～10ml) solution of water 3.92mmol).Mixture is in reflux 3 hours, cools off then and be evaporated to stay minimum water.It is acidified to pH1 with the 2M aqueous hydrochloric acid.The water layer washed with dichloromethane separates and evaporates obtaining 1-methyl-5-oxo-proline(Pro) then, is white solid (0.300g).
Embodiment 121-ethyl-5-oxo-N-[(2,3, the 4-trifluorophenyl) methyl]-prolineamide (E12)
With 1-ethyl-5-oxo-proline(Pro) (0.050g, 0.32mmol) be dissolved in anhydrous methylene chloride (～7ml) and in the dimethyl formamide (1ml), and to wherein adding I-hydroxybenzotriazole (0.052g, 0.38mmol), [(2,3,4-trifluorophenyl) methyl] and amine (0.103g, 0.64mmol), N-ethylmorpholine (0.151ml, 0.95mmol) and N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.073g, 0.38mmol).Mixture is spent weekend in the room temperature jolting.(0.051g 0.32mmol) is added in the mixture, and continues to stir the longer time and show do not have further product to generate up to HPLC with [(2,3, the 4-trifluorophenyl) methyl] amine of another equal portions.Mixture filters by phase splitter then with 2M aqueous hydrochloric acid (5ml) dilution.Organic layer washs with saturated sodium bicarbonate aqueous solution, and filters by phase splitter once more.Evaporate organic layer then and obtain crude product.Thick material obtains pure 1-ethyl-5-oxo-N-[(2,3,4-trifluorophenyl by the automatic HPLC purifying of mass spectrometric detection) methyl]-prolineamide (0.032g).
LC/MS [M+H]
+=301, retention time=2.03 minute.
1-ethyl-5-oxo-the proline(Pro) that uses in the aforesaid method is prepared as follows (method A):
(i) (5.0g 23.7mmol) is dissolved in the methyl alcohol (100ml), and (1.0g 24.9mmol) handles mixture with ground sodium hydroxide in room temperature under argon gas then with L-glutamic acid dimethyl ester hydrochloride.After 5 minutes, (1.99ml 35.5mmol) also continues to stir 10 minutes to add acetaldehyde.Mixture is cooled to 0 ℃, and (0.701g handles 18.95mmol) with the sodium borohydride particle.Continue to stir 1 hour in 0 ℃, evaporate methyl alcohol then, and resistates is dissolved in ethyl acetate and filtration.Filtrate is used the salt water washing then, and this salt water lotion ethyl acetate extraction.The ethyl acetate fraction that merges is filtered by the hydrophobic glass material, and evaporation obtains clarifying oily matter (3.2g).Oily matter is dissolved in the toluene (30ml), and spends the night in reflux.Evaporate toluene then and obtain greenish orange look resistates, it is by quick silica gel column chromatography, comes purifying with the hexane solution wash-out of the ethyl acetate of gradient 20-60%, obtains partial-purified 1-ethyl-5-oxo-proline methyl ester (1.9g), is to clarify oily matter.It is not further purified and just is used for next step.
(ii) (1.91g 11.17mmol) is dissolved in the methyl alcohol (25ml), and (7.3ml 14.52mmol) handles with the 2M aqueous sodium hydroxide solution with 1-ethyl-5-oxo-proline methyl ester.Mixture is used washed with dichloromethane then in stirring at room 4 hours.The evaporation water layer, and resistates is with the (～5ml) processing of the ethereal solution of excessive 1M hydrogenchloride.Evaporating mixture once more, and resistates grinds with methylene dichloride.Remove solid matter, and the methylene dichloride fraction of evaporation merging, obtain clarifying oily matter, leave standstill crystallization.Grind with hexane and ether, and drying obtains pure 1-ethyl-5-oxo-proline(Pro) (0.271g), be white solid.
Perhaps, 1-ethyl-5-oxo-proline(Pro) can be prepared as follows (method B):
(i) with 5-oxo-L-proline(Pro) 1,1-dimethyl ethyl ester (2.7g, 12mmol, as Synth.Comm., preparation in 2005,35 (8), 1129) is added to sodium hydride (0.428g (60% in oil suspension), 10.7mmol) in the suspension in tetrahydrofuran (THF) (6ml), and mixture was in stirring at room 5 minutes.(1.67g, 10.7mmol), and mixture was in 40 ℃ of heating 2 hours to add iodoethane then.Add other sodium hydrides (0.24g), and continue stirring in room temperature and spend the night.At this moment, again another part iodoethane (0.86ml) is added in the mixture.Mixture was placed weekend in room temperature.With water (～10ml) be added in the mixture, and it was stirred 15 minutes.The evaporation tetrahydrofuran (THF), and remaining water layer is with methylene dichloride (2 * 50ml) and the extraction of 3: 1 mixtures (50ml) of chloroform and Virahol.The organic phase that merges is filtered by the hydrophobic glass material, and evaporation obtains yellow oil.Toluene is added in the mixture, and evaporation obtains yellow oil once more.This material is by automatic flash chromatography on silica gel (Biotage SP4), comes purifying with the hexane solution wash-out of the ethyl acetate of 15-100% gradient, obtains pure 1-ethyl-5-oxo-proline(Pro) 1,1-dimethyl ethyl ester.
(ii) with 1-ethyl-5-oxo-proline(Pro) 1,1-dimethyl ethyl ester (0.965g) be dissolved in methylene dichloride (～5ml) in, and handle with trifluoroacetic acid (1ml).Mixture is in stirring at room 1.5 hours, evaporation then.Gained material major part is an initial substance, therefore add again trifluoroacetic acid (1ml) and methylene dichloride (～5ml), and in stirring at room mixture 36 hours.Evaporating mixture is added to toluene in the resistates then, once more evaporation.Repeat this process once more, obtain thick 1-ethyl-5-oxo-proline(Pro), be dark yellow oily thing, it does not have repurity promptly to use.
Perhaps, 1-ethyl-5-oxo-proline(Pro) can be prepared as follows (method C):
(i) with 5-methyl-L-L-glutamic acid 1-(1, the 1-dimethyl ethyl) ester hydrochloride (5.0g, 19.71mmol) be dissolved in the mixture of methyl alcohol (30ml) and tetrahydrofuran (THF) (60ml), then under argon gas under room temperature, with sodium hydroxide (0.828g, 20.69mmol) processing of mixture with the powdered of pulverizing.Stir after 10 minutes, add acetaldehyde (1.11ml, 19.71mmol) and acetate (1.13ml 19.71mmol), and continues stirring 10-15 minute.Mixture is cooled to 0 ℃ in ice bath, and (0.746g handles 19.71mmol) with the sodium borohydride sheet.Continue under 0 ℃ of argon gas and stirred～1 hour.Mixture is warmed to room temperature obtains dense suspension.Leach the white solid of particulate, evaporate methyl alcohol then, and with resistates be dissolved in methylene dichloride (～50ml) in, and with saturated sodium bicarbonate aqueous solution (～25ml) washing.Use phase splitter to separate organic layer, water layer is used methylene dichloride (2 * 20ml) reextraction (back extract) more then.The organic layer that evaporation merges obtain colorless oil (～4g).This oily matter (3g supposes 14.8mmol) is dissolved in the toluene (30ml), and in the heating down～16 hours that refluxes, spending the night obtains orange solution.Evaporate toluene then and obtain orange (2.6g).Itself and another batch oily matter (0.850g) that uses same procedure to obtain merges, then by automatically quick silica gel column chromatography (Biotage SP4), hexane solution wash-out purifying with gradient 20-80% ethyl acetate, obtain pure 1-ethyl-5-oxo proline(Pro) 1,1-dimethyl ethyl ester (2.14g).
(ii) with 1-ethyl-5-oxo proline(Pro) 1,1-dimethyl ethyl ester (0.933g) be dissolved in methylene dichloride (～5ml) in, and handle with trifluoroacetic acid (1ml).Mixture is in stirring at room 3 hours, evaporation then.Resistates is dissolved in the toluene, and evaporation once more.Obtain the 1-ethyl-5-oxo-proline(Pro) of part pure (＞95%), be orange/yellow oil (0.914g), it is not further purified promptly and uses.
Embodiment 13-36
With with the foregoing description 12 similar methods, prepare the compound in the following table (table 2) by [(2,3, the 4-trifluorophenyl) methyl] amine that replaces with suitable amine (or its salt) using in the aforesaid method.All used amine of compound shown in the preparation table 2 maybe can prepare by the approach of having described in the chemical literature or prepare by method similar with it from commercially available.In every kind of situation, the preparation of method shown in used 1-ethyl-5-oxo-proline(Pro) passes through in the reaction.When detecting (by chirality HPLC), the enantiomer of isomer shown in also having listed excessive (e.e.), and stereospecificity title, corresponding retention time (r.t.) in used chiral separation method and this method.
Table 2
Synthetic N-[(2-chloro-3,4-difluorophenyl) methyl]-required [(2-chloro-3, the 4-difluorophenyl) methyl] amine hydrochlorate of 1-ethyl-5-oxo prolineamide (embodiment 36) prepares with following method:
(i) with N, N, N ', N '-Tetramethyl Ethylene Diamine (39.6ml, tetrahydrofuran (THF) 264mmol) (170ml) solution is cooled to-70 ℃ under argon gas, add then s-butyl lithium (205ml, 288mmol).Through 40 minutes, add 3 in mixture then, (19g, tetrahydrofuran (THF) 120mmol) (80ml) solution guarantee that mixture temperature is not higher than-60 ℃ to the 4-difluoro-benzoic acid.Mixture stirred 1 hour in-68 ℃ to-70 ℃ temperature then, and (100g, tetrahydrofuran (THF) 422mmol) (170ml) solution keep the temperature of mixture to be lower than-60 ℃ simultaneously to add hexachloroethane through 35 minutes then.Mixture stirred 2 hours in-65 ℃ to-70 ℃ temperature.Mixture is warmed to-10 ℃, adds entry (500ml) cancellation reaction then.Mixture is with ether (250ml) dilution, and separating obtained two-layer.It is 1 that water layer is acidified to pH with concentrated hydrochloric acid aqueous solution, uses the extracted with diethyl ether of 2 * 500ml equal portions then.The organic extract liquid that merges is by the hydrophobic glass material, and the concentrated in a vacuum yellow solid that obtains.Its recrystallization from ethyl acetate obtains the pure 2-chloro-3 of two batches (8.35g and 4.47g), 4-difluoro-benzoic acid.
(ii) with 2-chloro-3, (2g 10.4mmol) handles with thionyl chloride (3.04ml) the 4-difluoro-benzoic acid, with mixture heating up to 80 ℃, keeps 90 minutes.Cooling mixture then, and concentrate in a vacuum.It is anhydrous 1 that resistates is dissolved in, and in the 4-diox (10ml), mixture cools off in ice-water bath then.(aqueous solution 25ml) drops in the mixture, with after it was warmed to 22 ℃ in 2 hours with 0.88 ammoniacal liquor.Use the 2-chloro-3 of 10.8g, 0.88 ammoniacal liquor of the thionyl chloride of 4-difluoro-benzoic acid, 8.2ml and 45ml repeats this process, merges two parts of mixtures then, and distributes in ethyl acetate (150ml) and water (100ml).Separate water layer and with the ethyl acetate extraction of 2 * 150ml equal portions.The organic extract liquid that merges then uses hydrophobic glass material drying with saturated sodium bicarbonate aqueous solution (100ml) washing, and vacuum concentration obtains 2-chloro-3, and 4-difluorobenzamide (11.86g) is white solid.
LC/MS [M+H]
+=192/194, retention time=1.69 minute.
(iii) with 2-chloro-3, (11.85g 62mmol) is dissolved in the tetrahydrofuran (THF) (200ml) the 4-difluorobenzamide, and (247ml 247mmol) handles with 1M borine tetrahydrofuran (THF).With mixture heating up to 70 ℃, and stirred 18 hours.Then mixture is cooled off in ice-water bath, and drip concentrated hydrochloric acid aqueous solution (150ml).And then in 70 ℃ the heating and stirred 2 hours.Cooling mixture, and vacuum evaporating solvent then.Resistates distributes between ethyl acetate (200ml) and 2N aqueous hydrochloric acid (200ml).Separate water layer, and regulate pH to 8-9 by Dropwise 5 N aqueous sodium hydroxide solution.(4 * 200ml) extractions, the organic extract liquid of He Binging is by the hydrophobic glass material then, and is concentrated into volume and is～200ml with ethyl acetate for the muddy suspension of gained.Mixture forms precipitation by adding diethyl ether solution (100ml) acidifying of 1M hydrogenchloride then.Vacuum evaporating solvent obtains white solid.This solid of recrystallization obtains 3 parts of pure [(2-chloro-3,4-difluorophenyl) methyl] amine hydrochlorates (merging quality=4.46g), be white solid from methylated spirit (60ml).
Embodiment 37N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-5-oxo-4-(phenyl methyl)-prolineamide (E37)
With thick 1-ethyl-5-oxo-4-(phenyl methyl)-proline(Pro) (0.052g, 0.09mmol, be prepared as follows) be suspended in the mixture of methylene dichloride (0.5ml) and dimethyl formamide (0.5ml), and (0.034ml 0.27mmol) makes most of substance dissolves to wherein adding N-ethylmorpholine.Add I-hydroxybenzotriazole (0.016g then, 0.12mmol) and N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.022g 0.12mmol), and stirred the mixture 10 minutes, add then [(2-chloro-4-fluorophenyl) methyl] amine (0.019g, 0.12mmol).Then mixture is placed in room temperature and spent the night.With saturated sodium bicarbonate aqueous solution (～2ml) be added in the mixture, and stirred 10 minutes.Organic layer is by the phase splitter filtering separation, then with the washing of 2M aqueous hydrochloric acid.Separate organic layer once more and evaporate obtaining yellow oil, its automatic HPLC purifying by mass spectrometric detection obtains pure N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-5-oxo-4-(phenyl methyl)-prolineamide (0.004g), be colorless oil.LC/MS[M+H]
+=389, retention time=2.90 minute.
The 1-ethyl that uses in the aforesaid method-5-oxo-4-(phenyl methyl)-proline(Pro) is prepared as follows (method A):
(i) with (S)-(+)-2-Pyrrolidone-5-methyl-formiate (0.85g 5.94mmol) is dissolved in the methylene dichloride (5ml), and with triethylamine (0.869ml, 6.24mmol) and 4-dimethylaminopyridine (0.010g) handle.(1.36g 6.24mmol), and stirs the gained orange solution and spends the night to wherein adding coke acid di-t-butyl ester (di-tertbutyl dicarbonate).Mixture becomes indigo plant/grey, and evaporating solvent obtains light gray oily matter (1.4g).It is by automatically quick silica gel column chromatography (Biotage SP4), hexane solution wash-out purifying with gradient 0-60% ethyl acetate obtains 2-methyl-5-oxo-1,2-tetramethyleneimine dioctyl phthalate 1-(1, the 1-dimethyl ethyl) ester (1.37g), be colorless oil, leave standstill crystallization.
(ii) with 2-methyl-5-oxo-1, (0.324g 1.33mmol) is dissolved in the tetrahydrofuran (THF) (3ml) 2-tetramethyleneimine dioctyl phthalate 1-(1, the 1-dimethyl ethyl) ester, and uses acetone/cardice bath mixture to be cooled to-78 ℃ under argon atmospher.The tetrahydrofuran (THF) of dropping 1M two (trimethyl silyl) lithium amide (1.4ml, 1.40mmol) solution, and under argon gas, stirred 1 hour.(0.174ml, 1.46mmol), and mixture was in-78 ℃ of restir 2.5 hours to wherein adding bromotoluene then.Then mixture is warmed to room temperature, and (～5ml) cancellation is placed in room temperature then and is spent the night by adding saturated aqueous ammonium chloride.Separate organic layer, and water layer water (5ml) dilution again, and (3 * 10ml) extract with ethyl acetate.The organic layer that merges filters then through anhydrous sodium sulfate drying, and the concentrated yellow oil (0.700g) that obtains.It is by automatically quick silica gel column chromatography (Biotage SP4), hexane solution wash-out purifying with 0-35% gradient ethyl acetate, behind the evaporating solvent, obtain 2-methyl-5-oxo-4-(phenyl methyl)-1,2-tetramethyleneimine dioctyl phthalate 1-(1, the 1-dimethyl ethyl) ester is white solid (0.418g).
(iii) with 2-methyl-5-oxo-4-(phenyl methyl)-1,2-tetramethyleneimine dioctyl phthalate 1-(1, the 1-dimethyl ethyl) ester (0.415g 1.24mmol) is dissolved in 4M hydrogenchloride De diox (2ml) solution, and in stirring at room 2 hours.Evaporating solvent obtains colorless oil, leaves standstill crystallization and obtains 5-oxo-4-(phenyl methyl)-proline methyl ester, is cream-coloured/white solid (0.205g).It is used for next step without purifying.
(iv) (0.205g 0.88mmol) is dissolved in the tetrahydrofuran (THF) (2.5ml), and (0.077ml 0.97mmol) handles with iodoethane with 5-oxo-4-(phenyl methyl)-proline methyl ester.Mixture is cooled to 0 ℃ then, and (0.037g, 60% suspension in oil are handled 0.92mmol) with sodium hydride.In 0 ℃ stir 10-15 minute after, solution is warmed to room temperature, and restir 3.5 hours.(methylene dichloride (5ml) dilution is used in～2ml) processing to mixture subsequently with saturated aqueous ammonium chloride then.Organic layer by the hydrophobic glass material (with the methylene dichloride of other equal portions (2 * 5ml) washing water layer) filtering separation.The organic phase that evaporation merges obtain brown oil (～0.100g).It is by automatically quick silica gel column chromatography, hexane solution wash-out purifying with 0-100% gradient ethyl acetate, obtain partial-purified (～90% is pure) 1-ethyl-5-oxo-4-(phenyl methyl)-proline methyl ester (0.024g), be yellow oil, it is not further purified and just is used for next step.
(v) (0.024g 0.09mmol) is dissolved in methyl alcohol (0.5ml), and is cooled to 0 ℃ in ice bath with 1-ethyl-5-oxo-4-(phenyl methyl)-proline methyl ester.(0.137ml 0.27mmol) is added in the mixture, and continues to stir 3 hours in 0 ℃ with the 2M aqueous sodium hydroxide solution.Evaporating solvent, and resistates by with the 2M aqueous hydrochloric acid (～0.2ml) handle acidifying to obtain turbid solution.Evaporation obtains thick 1-ethyl-5-oxo-4-(phenyl methyl)-proline(Pro) (0.052g), is the mixture of white solid and yellow oily resistates.It is not further purified promptly and uses.
Perhaps 1-ethyl-5-oxo-4-(phenyl methyl)-proline(Pro) also can prepare (method B) with following method:
(i) (1.88g 20mmol) is dissolved in the dimethyl formamide (9ml), and (60% suspension in oil, 0.220g 5.5mmol) handle with sodium hydride with (S)-(+)-L-5-trimethylphenyl oxygen ylmethyl-2-Pyrrolidone in 0 ℃.Mixture was in 0 ℃ of stirring 30 minutes, and (0.444ml 5.5mmol) handles to use iodoethane then.Mixture is warmed to room temperature, stirs then and spend the night.Then mixture is distributed in ethyl acetate and saturated aqueous ammonium chloride, and extract with ethyl acetate (x3).The organic extract liquid that merges is water successively, and 50% sodium chloride aqueous solution (x2) and saturated sodium-chloride water solution washing are then through dried over sodium sulfate.Concentrate and obtain beige solid, it is by automatically quick silica gel column chromatography (Biotage SP4), and the hexane solution wash-out purifying with 0-100% gradient ethyl acetate obtains pure 1-ethyl-5-{[(trityl group) the oxygen base] methyl }-2-Pyrrolidone (1.78g).
(ii) with the tetrahydrofuran (THF) (1.050ml of 2M diisopropylaminoethyl lithium, 2.1mmol) solution joins 1-ethyl-5-{[(trityl group in-78 ℃) the oxygen base] methyl-2-Pyrrolidone (0.771g, in tetrahydrofuran (THF) 2mmol) (10ml) solution, and stirred the gained mixtures 1 hour in-78 ℃.(0.262ml 2.2mmol), and after 1 hour, is warmed to ambient temperature overnight with mixture in-78 ℃ of stirrings again to add bromotoluene then.Ethyl acetate (x3) extraction is used in mixture saturated aqueous ammonium chloride cancellation then.The organic extract liquid of He Binging washes with water then, uses saturated sodium-chloride water solution (x2) washing then, through anhydrous magnesium sulfate drying, and the concentrated thick oily matter (1.27g) that obtains.Thick solid is by automatically quick silica gel column chromatography (Biotage SP4), the hexane solution wash-out purifying of 0-100% gradient ethyl acetate, obtain required product (being 1-ethyl-3-(phenyl methyl)-5-{[(trityl group) oxygen base] methyl }-2-Pyrrolidone (0.561g)), it is used for next step, and unreacted starting materials and dialkyl group product, 1-ethyl-3,3-two (phenyl methyl)-5-{[(trityl group) the oxygen base] methyl }-2-Pyrrolidone (0.053g).
(iii) 1-ethyl-3-(phenyl methyl)-5-{[(trityl group) oxygen base] methyl }-(0.561g 1.1mmol) stirred 24 hours in the mixture of acetonitrile (21ml) and formic acid (3ml) in room temperature 2-Pyrrolidone.At this moment reaction not fully, so evaporating solvent and change, and continue to stir 3 hours with formic acid (10ml).Reaction still not fully, therefore enriched mixture (removing all formic acid with methanol azeotropic) in a vacuum is dissolved in methyl alcohol (20ml) then.Then with Amberlyst
Join in the mixture, and continue stirring in room temperature and spend the night.Leach resin, use methanol wash, concentrated filtrate to obtain jelly (0.625g) again.This jelly is by automatically quick silica gel column chromatography (Biotage SP4), hexane solution wash-out purifying with 0-100% gradient ethyl acetate, obtain 1-ethyl-5-(hydroxymethyl)-3-(phenyl methyl)-2-Pyrrolidone (0.170g), it is used for next step.
(iv) with 1-ethyl-5-(hydroxymethyl)-3-(phenyl methyl)-2-Pyrrolidone (0.748g, 3.21mmol) be dissolved in acetonitrile (5ml) and 1M SODIUM PHOSPHATE, MONOBASIC aqueous buffer solution (3.69ml, 3.69mmol) in, the TEMPO (2 that adds a small amount of crystalline state, 2,6,6-tetramethyl--piperidino oxygen base free radical) and Textone (0.580g, 6.41mmol), and mixture is warmed to 40 ℃.Then about 1 SYNTHETIC OPTICAL WHITNER (chlorine bleach liquor, available chlorine＞12%) is added in the mixture, and continues to stir 3 hours in 40 ℃.Then mixture is poured into the ice that contains the 1%w/w S-WAT-waterborne, and the gained mixture is adjusted to pH2, use ethyl acetate (x3) extraction then with the 5N aqueous hydrochloric acid.The organic extract liquid that merges washs with saturated sodium-chloride water solution, then through dried over mgso, and concentrates and obtains 1-ethyl-5-oxo-4-(phenyl methyl) proline(Pro) (0.807g), is solid, and it is not further purified promptly and uses.
Embodiment 38N-[(2-chloro-4-fluorophenyl) methyl]-1-(2-methyl-2-propylene-1-yl)-5-oxo prolineamide (E38)
With thick 1-(2-methyl-2-propylene-1-yl)-5-oxo proline(Pro) (～0.075g,～0.41mmol, be prepared as follows) be dissolved in the methylene dichloride (5ml), and to wherein adding I-hydroxybenzotriazole (0.061g, 0.45mmol), [(2-chloro-4-fluorophenyl) methyl] amine (0.068g, 0.43mmol) and N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.087g, 0.45mmol).Mixture was in stirring at room 24 hours then.Mixture is again with the methylene dichloride dilution, then successively with 2M aqueous hydrochloric acid and saturated sodium bicarbonate aqueous solution washing.Organic layer filters by phase splitter, and evaporation obtains the brown resistates, its automatic HPLC purifying by mass spectrometric detection obtains pure N-[(2-chloro-4-fluorophenyl) methyl]-1-(2-methyl-2-propylene-1-yl)-5-oxo prolineamide (0.018g), be white solid.LC/MS[M+H]
+=325.1, retention time=2.40 minute.
1-(2-methyl-2-propylene-1-yl)-5-oxo proline(Pro) that aforesaid method uses is prepared as follows:
(i) methyl alcohol (55ml) is cooled to-10 ℃ (using the cardice/ tetracol phenixin to bathe), stirs simultaneously, then through 45 minutes thionyl chloride.(10g, 67.96mmol), stirring reaction is 3 hours then, is warmed to 21 ℃ simultaneously to divide 3 parts of warp～5 minute adding (D)-L-glutamic acid then.Vacuum evaporating solvent obtains clarifying oily matter (15g), in the mixture with its water-soluble (150ml) and diox (150ml).(46g 340mmol), and stirs to wherein slowly adding yellow soda ash then.(9.64ml 68mmol), and continues to stir and spends the night to add benzyl chloroformate then.Mixture carefully uses 2N aqueous hydrochloric acid (250ml) to handle, and uses ethyl acetate (2 * 250ml) extractions then.The organic fraction salt water washing that merges, dry then and evaporation obtains clarifying oily matter (18.7g).It is dissolved in the methylene dichloride (400ml), and handles with the vitriol oil (1ml).Then a large amount of excessive iso-butylenes are concentrated in the mixture, and spend the night in 21 ℃ of stirrings.Then with saturated sodium bicarbonate aqueous solution (～400ml) carefully join in the mixture, separate organic phase then, use the salt water washing, dry and vacuum-evaporation obtains clarifying oily matter (21.9g).It is by silica gel column chromatography, obtains pure 1-(1, the 1-dimethyl ethyl) 5-methyl N-{ [(phenyl methyl) oxygen base] carbonyl } glutamate (4.87g) with the mixture wash-out purifying of 3: 1 hexanaphthenes and ethyl acetate.
(ii) minute clockwise hexamethyldisilane base amine potassium (potassoimhexamethyldisilazide) (10ml in-70 ℃ of warp～5,0.6M toluene solution, drip 1-(1 in tetrahydrofuran (THF) 6mmol) (25ml) solution, the 1-dimethyl ethyl) 5-methyl N-{ [(phenyl methyl) oxygen base] carbonyl } glutamate (1.05g, tetrahydrofuran (THF) 3mmol) (10ml) solution.Mixture is used methylallyl iodate thing (2.18g, tetrahydrofuran (THF) 12mmol) (10ml) solution-treated then in-70 ℃ of stirrings 1 hour.Continue to stir 2 hours in-78 ℃, be warmed to 21 ℃ then.Behind the restir 1 hour, mixture is poured in the 1N aqueous hydrochloric acid, and (2 * 50ml) extract with ethyl acetate.The organism salt water washing that merges, drying, and vacuum-evaporation obtains yellow oil (1.03g).It is by silica gel column chromatography, obtains pure 1-(2-methyl-2-propylene-1-yl)-5-oxo proline(Pro) 1 with the mixture wash-out purifying of 4: 1 hexanaphthenes and ethyl acetate, and 1-dimethyl ethyl ester is to clarify oily matter (0.322g).
(iii) with 1-(2-methyl-2-propylene-1-yl)-5-oxo proline(Pro) 1,1-dimethyl ethyl ester (0.099g 0.41mmol) is dissolved in the mixture of methylene dichloride (2ml) and trifluoroacetic acid (2ml), and in stirred overnight at room temperature.Evaporating solvent (removing the trifluoroacetic acid of trace with methylbenzene azeotropic) obtains thick 1-(2-methyl-2-propylene-1-yl)-5-oxo proline(Pro), is brown oil, and it is not further purified promptly and uses.
Embodiment 391-cyclopropyl-N-[(2, the 4-dichlorophenyl) methyl]-5-oxo prolineamide (E39)
(0.047g, 0.25mmol) (15% aqueous solution, 0.26ml add cyclopropylamine (0.042ml, 0.6 mmol) in methyl alcohol 0.4mmol) (1.75ml) solution with the 4-ketobutyric acid to (2,4 dichloro benzene base) methyl isocyanide.Mixture is heated to 100 ℃ in microwave reactor, kept 30 minutes.Vacuum removes solvent, and the automatic HPLC purifying of resistates by mass spectrometric detection obtains 1-cyclopropyl-N-[(2, the 4-dichlorophenyl) methyl]-5-oxo prolineamide (0.072g), be white solid.LC/MS[M+H]
+=326/328, retention time=2.29 minute.
Embodiment 40N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-cyclopropyl-5-oxo prolineamide (E40)
To [2-chloro-3-(trifluoromethyl) phenyl] methyl isocyanide (0.088g, 0.4mmol) and succinic acid semialdehyde (15% aqueous solution, 0.26ml, add in methyl alcohol 0.4mmol) (1.75ml) solution cyclopropylamine (0.042ml, 0.6mmol).Mixture is heated to 100 ℃ in microwave reactor, kept 30 minutes.Vacuum removes solvent, and the automatic HPLC purifying of resistates by mass spectrometric detection obtains N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-cyclopropyl-5-oxo prolineamide (0.076g), be white solid.LC/MS[M+H]
+=361/363, retention time=2.39 minute.
[2-chloro-3-(trifluoromethyl) phenyl] the methyl isocyanide that uses in the aforesaid method is prepared as follows:
(i) (1.05g, anhydrous tetrahydro furan 5mmol) (10ml) drips of solution is added to N-formyl radical benzotriazole, and (0.772g is in anhydrous tetrahydro furan 5.25mmol) (10ml) solution with { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine.React on 22 ℃ and stirred 18 hours, vacuum concentration, and resistates then is allocated between methylene dichloride (75ml) and the 2N aqueous sodium hydroxide solution (40ml).Separate organic layer, and extract with 2N aqueous sodium hydroxide solution (40ml).Organic layer is by the hydrophobic glass material, and vacuum concentration obtains white solid.Crude product is by automatic fast silica gel column chromatography (Biotage SP4), obtains { [2-chloro-3-(trifluoromethyl) phenyl] methyl } methane amide with the dichloromethane solution wash-out purifying of gradient 0-10% ethyl acetate, is white solid.
(ii) argon gas is down with { [2-chloro-3-(trifluoromethyl) phenyl] methyl } methane amide (0.67g, 2.82mmol) anhydrous methylene chloride (20ml) solution in ice-water bath, cool off, add diisopropylamine (1.78ml then, 12.7mmol), add then phosphoryl chloride (0.393ml, 4.23mmol).Reacting on 2-5 ℃ stirred 2 hours.Mixture is at vacuum concentration then, and resistates handles with saturated sodium bicarbonate aqueous solution (20ml), and extracts with methylene dichloride (20ml).Organic layer is by the hydrophobic glass material, and vacuum concentration obtains yellow solid.Further vacuum-drying obtains [2-chloro-3-(trifluoromethyl) phenyl] methyl isocyanide, is orange jelly (0.66g), and it is not further purified promptly and uses.
Embodiment 41N-[(2-chloro-4-fluorophenyl) methyl]-1-cyclopropyl-5-oxo prolineamide (E41)
To [2-chloro-4-fluoro-phenyl] methyl isocyanide (0.068g, 0.4mmol) and succinic acid semialdehyde (15% aqueous solution, 0.26ml, 0.4mmol) add in the solution in methyl alcohol (1.75ml) cyclopropylamine (0.042ml, 0.6mmol).Mixture is heated to 100 ℃ in microwave reactor, kept 30 minutes.Vacuum removes solvent, and the automatic HPLC purifying of resistates by mass spectrometric detection obtain colourless jelly, and it grinds with ether and obtains N-[(2-chloro-4-fluorophenyl) methyl]-1-cyclopropyl-5-oxo prolineamide, be oldlace solid (0.058g).LC/MS[M+H]
+=310, retention time=2.16 minute.
As [2-chloro-4-fluoro-phenyl] methyl isocyanide of initial substance with embodiment 40 in the described similar methods preparation of preparation [2-chloro-3-(trifluoromethyl) phenyl] methyl isocyanide, except using 2-chloro-4-fluorophenyl] methyl amine replaces 2-chloro-3-(trifluoromethyl) phenyl] methyl amine.
Embodiment 42N-[(2, the 4-dichlorophenyl) methyl]-1-ethyl-4,4-dimethyl-5-oxo prolineamide (E42)
(0.075g, 0.4mmol) and thick 2, (0.115g 0.6mmol) is dissolved in the methyl alcohol (2ml) 2-dimethyl-4-ketobutyric acid with (2,4 dichloro benzene base) methyl isocyanide.(0.6mmol), and mixture heated 30 minutes in 100 ℃ in sealed tube in microwave reactor for the 2M aqueous solution, 0.3ml to add ethylamine solution.Mixture was placed weekend, vacuum removes solvent then, and the gained orange obtains clarifying the oily jelly by the automatic HPLC purifying of mass spectrometric detection, it grinds with ether and obtains N-[(2, the 4-dichlorophenyl) methyl]-1-ethyl-4,4-dimethyl-5-oxo prolineamide is white solid (0.024g).LC/MS[M+H]
+=343, retention time=2.57 minute.
Be used as 2 of initial substance in the aforesaid method, 2-dimethyl-4-ketobutyric acid is prepared as follows:
(i) at CO
2With 2,2-dimethyl-4-pentenoic acid is dissolved in methylene dichloride (25ml) in/acetone bath, and is cooled to-78 ℃, and blasts oxygen in mixture, keeps 5 minutes.Open ozonizer, in mixture, blast ozone, kept 15 minutes.Stop flow of ozone then, and mixture was used argon purge 2 minutes then with oxygen blow 5 minutes.TLC shows that reaction obviously do not carry out, and therefore blasts ozone again 15 minutes in mixture, keeps light blue afterwards and forms suspension.Turn off flow of ozone, and mixture was used argon purge 10 minutes (becoming negative for wet starch/iodine test paper up to waste gas) then with oxygen blow 5 minutes.(1.72ml 23.41mmol) joins in the mixture, then mixture is warmed to room temperature with dimethylsulphide then.After 2 hours, enriched mixture obtains colorless oil (1.5g) in stirring at room.1.4g this material is by quick silica gel column chromatography, comes purifying with the dichloromethane solution wash-out of gradient 0-50% ethyl acetate, obtains 2,2-dimethyl-4-ketobutyric acid is colorless oil (0.649g).
Embodiment 43N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-(1-methylethyl)-5-oxo prolineamide (E43)
With 1-(1-methylethyl)-5-oxo proline(Pro) (0.100g, 0.58mmol) be dissolved in the methylene dichloride (20ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.111g, 0.58mmol), I-hydroxybenzotriazole (0.078g, 0.58mmol) and N-ethylmorpholine (0.223ml, 1.75mmol).{ [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine is added to mixture the most at last, and continues to stir～48 hours.Mixture is handled with saturated sodium bicarbonate aqueous solution (20ml) then, and vigorous stirring.Water layer uses phase splitter to remove, and uses the argon purge element to remove solvent from organic layer then.Gained resistates water and ethyl acetate (25ml, 1: 1) are handled, and discard water layer subsequently.Organic layer filters by phase splitter, and evaporation obtains oily matter.It grinds with ether and obtains solid, and its automatic HPLC purifying with mass spectrometric detection obtains N-{[2-chloro-3-(trifluoromethyl) phenyl subsequently] methyl }-1-(1-methylethyl)-5-oxo prolineamide (0.097g), be white solid.LC/MS[M+H]
+=363, retention time=2.48 minute.
The 1-that uses in the aforesaid method (1-methylethyl)-5-oxo proline(Pro) is to prepare with similar methods described in synthetic 1-ethyl-5-oxo-proline methyl ester (seeing embodiment 3); except using acetone to replace acetaldehyde; and add ester subsequently and carry out deprotection steps (use standard conditions, the i.e. methanol solution of sodium hydroxide) (opposite with the acid amides coupling) with the deprotection of the associating described in the embodiment 3.
Embodiment 44-49
With with embodiment 43 described similar approach, by the compound in { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine preparation table (table 3) that replaces with suitable amine (or its salt) using in the aforesaid method.Unless otherwise indicated, all used amine of the compound shown in the preparation table 3 maybe can use the method described in the chemical literature to prepare from commercially available.
Table 3
Synthetic N-[(2,3-two chloro-4-fluorophenyls) methyl]-required [(2, the 3-two chloro-4-fluorophenyls) methyl] amine hydrochlorate of 1-(1-methylethyl)-5-oxo-L-prolineamide (embodiment 48) prepares with following method:
(i) (0.172g, (0.400g is in (20ml) solution of water 2.5mmol) and 37% aqueous hydrochloric acid (5ml) 2.5mmol) to be added to the 2-chloro-6-fluoro-3-methyl-phenyl amine of stirring with Sodium Nitrite in-5 ℃.Mixture was in-5 ℃ of stirrings 5 minutes, and (0.742g, 7.5mmol) in the solution in 37% aqueous hydrochloric acid (5ml), keeping temperature simultaneously is-5 to 0 ℃ once to join cuprous chloride (I) then.Reaction mixture is heated to 38 ℃ and stirred 1 hour,, and adds ether (20ml) then with the mixture cooling.Separate organic phase, and, wash with water then with the washing of 1N aqueous hydrochloric acid.Organic layer is through dried over sodium sulfate then, and vacuum concentration.Thick resistates with sherwood oil wash-out purifying, obtains 2 by quick silica gel column chromatography, and (0.090g 0.5mmol), is white solid to 3-two chloro-1-fluoro-4-methylbenzene.
(ii) with 2, (0.090g, (0.284g is 1mmol) in the mixture in acetate (1ml) 0.5mmol) to be added to the potassium bichromate of stirring for 3-two chloro-1-fluoro-4-methylbenzene.Then 97% sulfuric acid (0.5ml) slowly is added to mixture, subsequently in 100 ℃ of heating 2 hours.After being cooled to room temperature, add entry and ice, leach the gained green solid, and use cold water washing, obtain 2, (0.056g 0.27mmol), is white solid to 3-two chloro-4-fluorobenzoic acids.
(iii) under the argon gas room temperature, with 2,3-two chloro-4-fluorobenzoic acid (0.200g, 0.92mmol) methylene dichloride (～4ml) solution is with I-hydroxybenzotriazole (0.162g, 1.2mmol), N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.230g, 1.2mmol) and triethylamine (0.56ml, 4.0mmol) processing.Mixture uses 32% ammonium hydroxide aqueous solution (0.088ml) to handle in stirring at room 40 minutes then, and in stirred overnight at room temperature.Mixture dilutes with methylene dichloride, and water and saturated sodium bicarbonate aqueous solution washing successively.Separate organic layer and through dried over sodium sulfate, concentrate then and obtain 2,3-two chloro-4-fluorobenzamides (0.156g) are white solid, and it is not further purified promptly and uses.
(iv) with 2, (0.750g, 3.62mmol) solution in anhydrous tetrahydro furan (2ml) is heated to 90 ℃ to 3-two chloro-4-fluorobenzamides under nitrogen.(1.05ml, 5.43mmol) solution is added in the hot solution, and continues to stir 4 hours with the tetrahydrofuran (THF) of 10M hydroborate dimethylsulphide mixture.Mixture is handled with the 6N aqueous hydrochloric acid then, and continues heating 30 minutes.Evaporating solvent, and thick resistates is by SCX post (cartridge) purifying, then by quick silica gel column chromatography, with the dichloromethane solution wash-out purifying of 5% methyl alcohol.Gained amine is handled with the ethereal solution of hydrogenchloride and is obtained [(2,3-two chloro-4-fluorophenyls) methyl] amine hydrochlorate (0.360g), is white solid.
Embodiment 50N-[(2, the 3-3,5-dimethylphenyl) methyl]-1-ethyl-5-oxo prolineamide (E50)
With 1-ethyl-5-oxo proline(Pro) (0.080g, 0.51mmol, with the described similar methods preparation of embodiment 12 method A) be dissolved in the methylene dichloride (5ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.117g, 0.61mmol), N-ethylmorpholine (0.195ml, 1.53mmol) and 2, the 3-dimethyl benzyl amine (0.082g, 0.61mmol).Stir the mixture～17 hours, placed weekend then.Then mixture with saturated sodium bicarbonate aqueous solution (～3ml) handle and vigorous stirring～10 minute.Use the hydrophobic glass material to separate organic layer, and water layer is used methylene dichloride (～2ml) extraction again.The organic layer that concentrate to merge obtain yellow oil (～0.2g).Further the automatic HPLC purifying by mass spectrometric detection obtains pure N-[(2, the 3-3,5-dimethylphenyl) methyl]-1-ethyl-5-oxo prolineamide (0.072g), be white solid.LC/MS[M+H]
+=275, retention time=2.12 minute.
Embodiment 51N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo prolineamide (E51)
With 1-methyl-5-oxo proline(Pro) (2.27g, 15.88mmol, be prepared as follows) be dissolved in the methylene dichloride (150ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (3.35g, 17.47mmol) and I-hydroxybenzotriazole (2.36g, 17.47mmol).Stir the mixture～10 minutes, (2.21ml, 15.88mmol) (3.66ml, 17.47mmol), and mixture is in stirred overnight at room temperature (～17 hours) with { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine to add triethylamine then.Form white precipitate during this.Then mixture with saturated sodium bicarbonate aqueous solution (～100ml) handle, and stirred 10 minutes.Use the hydrophobic glass material to separate organic layer, add the 2N aqueous hydrochloric acid then, and mix and separate once more.Concentrated organic layer obtain white solid (～2.5g).With this solid be dissolved in ethyl acetate (～200ml) and water (salt solution (50ml) washing is used in 4 * 50ml) washings then.Organic layer passes through the phase splitter drying then, and concentrated pure N-{[2-chloro-3-(trifluoromethyl) phenyl that obtains] methyl }-1-methyl-5-oxo-L-prolineamide, be tenderly white look solid (2.48g).
LC/MS[M+H]
+=335, retention time=2.24 minute.
1H NMR (CDCl
3, 500MHz) δ 2.02 (m, 1H), 2.35 (m, 1H), 2.39 (m, 1H), 2.47 (m, 1H), 2.81 (s, 3H), 4.00 (dd, 1H, J=8.9,4.2Hz), 4.60 (dd, 1H, J=15.1,6.2Hz), 4.65 (dd, 1H, J=15.1,6.2Hz), 6.56 (wide t, 1H, J=5.8Hz), 7.38 (t, 1H, J=7.7Hz), 7.60 (dd, 1H, J=7.6,1.0Hz), 7.68 (dd, 1H, J=7.9,1.2Hz); 13C NMR δ 176.0,171.5,137.5,133.9,131.7,129.3,127.4,127.0,122.8,63.8,41.8,29.4,29.2,23.4.
Prepare as the following method of the 1-methyl-5-oxo proline(Pro) of initial substance:
(i) with N-methyl-L-L-glutamic acid (9.81g 60.87mmol) is divided into 2 equal portions, and every part be suspended in the water (15ml), and in microwave reactor in sealed tube in 140 ℃ of heating 30 minutes, obtain settled solution.Merge two parts then, vaporize water and vacuum-drying obtain white solid.This solid grinds with ether and filters then and wash with ether, obtains 1-methyl-5-oxo-proline(Pro) (7.47g) after the drying, is white solid.
N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo prolineamide also can be prepared as follows:
1-methyl-5-oxo proline(Pro) (49.0g, 0.342mol, as above preparation) is suspended in DCM (600ml) (internal temperature is reduced to 13.7 ℃ from 20 ℃).Disposable adding EEDQ (2-oxyethyl group-1-ethoxy carbonyl-1,2-dihydroquinoline, 75.26g, 0.359mol, 1.05 equivalents), and mixture was in stirring at room 15 minutes.Then through 20 minutes with 1-[2-chloro-3-(trifluoromethyl) phenyl] methylamine (88.77g, 0.359mol, 1.05 DCM equivalent) (250ml) drips of solution adds to mixture (slight exotherm to 19 ℃), and uses DCM (150ml) that any residual solid is washed in the mixture again.Mixture is in stirred overnight at room temperature then.
Add saturated sodium bicarbonate aqueous solution (300ml), and in stirring at room mixture 5 minutes.Separate organic layer, and water (300ml) successively, (3 * 300ml), water (300ml) and saturated sodium-chloride water solution (300ml) wash the 2N aqueous hydrochloric acid.Organic solution is filtered through anhydrous sodium sulfate drying, and vacuum-evaporation.(～500ml) grinding, and collection solid obtain colorless solid (91.1g, 80%) with ether washing and dry (30 ℃, it's weekend is past vacuum drying oven) to the gained solid with ether then.This material and with similar batch of merging of similar approach preparation, and the material (178g altogether) that merges is dissolved in ethyl acetate (2.751), heating simultaneously (gentle reflux, stir on top).The hot settled solution of gentle agitation gained also is cooled to ambient temperature overnight.Collect solid, with cold ethyl acetate (500ml) washing, and dry (in 50 ℃ of vacuum drying ovens ,～3 days) obtain N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo prolineamide, be colourless needle-like (148.4g).
LC/MS[M+H]
+=335/337, retention time=2.26 minute.
1H NMR (CDCl
3, 500MHz) δ 1.86 (m, 1H), 2.21 (m, 1H), 2.24 (m, 1H), 2.28 (m, 1H), 2.64 (s, 3H), 4.12 (dd, 1H, J=8.3,3.5Hz), 4.47 (d, 2H, J=5.8Hz),, 7.58 (t, 1H, J=7.8Hz), 7.65 (dd, 1H, J=7.8,1.0Hz), 7.80 (dd, 1H, J=7.8,1.2Hz), 8.81 (wide t, 1H, J=5.7Hz);
13C NMR δ 174.4,171.4,138.8,133.1,129.8,127.5,127.1,126.6,122.9,61.6,40.2,29.1,28.0,22.5.
Enantiomer is excessive=and 99.1%, detect by chiral chromatography A, be indicated as N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo-L-prolineamide
Retention time=6.99 minute
[α]
D=-0.8 ° (c=1, MeOH), temperature=29.3 ℃, wavelength=589nm
Fusing point=173 ℃
Embodiment 52N-[(2,3-two chloro-4-fluorophenyls) methyl]-1-methyl-5-oxo prolineamide (E52)
1-methyl-5-oxo proline(Pro) (0.060g, 0.42mmol, as above embodiment 51 preparations) be dissolved in methylene dichloride (5ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.096g, 0.5mmol), I-hydroxybenzotriazole (0.068g, 0.5mmol) and N-ethylmorpholine (0.160ml, 1.26mmol).Stir the mixture～10 minutes, add [(2,3-two chloro-4-fluorophenyls) methyl] amine hydrochlorate (0.081g, 0.42mmol is as above-mentioned embodiment 48 preparations) then, and spend the night (～17 hours) that stir the mixture, make it spend weekend then.Mixture saturated sodium bicarbonate aqueous solution (～3ml) processing, and vigorous stirring 10 minutes then.Use the hydrophobic glass material to separate organic layer, use methylene dichloride (～2ml) washing water layer again.Concentrate the organic fraction that merges, obtain beige solid.This solids constituent fit over ethyl acetate (～20ml) and water (～10ml) between, separate organic layer by phase splitter then, and concentrate and obtain pure-[(2,3-two chloro-4-fluorophenyls) methyl]-1-methyl-5-oxo prolineamide, be pale solid.
LC/MS[M+H]
+=319, retention time=2.2 minute.
Embodiment 53-64
With with embodiment 52 similar methods, [(2, the 3-two chloro-4-fluorophenyls) methyl] amine hydrochlorate that replaces using in the aforesaid method with suitable amine (or its salt) prepares the compound in the following table (table 4).All used amine of compound shown in the preparation table 4 maybe can use the approach preparation described in the chemical literature or use similar method preparation from commercially available.
Table 4
The required amine of synthetic embodiment 62-64 prepares according to following method respectively:
1) { [2-methyl-3-(trifluoromethyl) phenyl] methyl } amine hydrochlorate (being used to prepare the amine of embodiment 62)
((2g in tetrahydrofuran (THF) 9.85mmol) (75ml) solution, and stirred 5 hours in 70 ℃ 39.4mmol) to be added to 2-methyl-3-trifluoromethyl benzamide for 1M, 39.4ml with the borine tetrahydrofuran (THF).LCMS shows that reaction is incomplete, therefore continues at heated overnight under 70 ℃ of argon gas, and then continues 5 hours.Reaction mixture is handled with the 2N aqueous hydrochloric acid, and in 100 ℃ of stirrings 4 hours, cools off weekend then.Vacuum concentrated mixture is allocated between methylene dichloride and the 2N aqueous sodium hydroxide solution then to doing.Use the hydrophobic glass material to separate organic layer, and the concentrated resistates that obtains, it is by quick silica gel column chromatography (with the dichloromethane solution wash-out of 0-5%2N ammonia/methyl alcohol) purifying.Evaporating solvent, and resistates is dissolved in ether, and handle with the ethereal solution of 1M hydrogenchloride.Filter the solid of collecting precipitation, grind with methylene dichloride then, obtain 2-methyl-3-(trifluoromethyl) phenyl after the filtration] methyl } amine hydrochlorate (1.4g), be white solid.
LC/MS[M+H]
+=173, retention time=1.30 minute.
2) [(2-bromo-4-fluorophenyl) methyl] amine hydrochlorate (amine that preparation embodiment 63 is used)
(i) (4g 21.6mmol) is incorporated in the dimethyl formamide (200ml), and stirs in 80 ℃ and to spend the night in 18 hours with 2-bromo-4-fluoro benzyl bromide (5g 18.8mmol) and phthalimide potassium.Vacuum concentrated mixture, and resistates is allocated between ether and the water.Leach solid, and water layer is used ether (2 * 50ml) washings again.Combined ether layer, and by dried over sodium sulfate filters then and evaporates and obtains pale solid (3.36g).Solid grinds with methyl alcohol, and filtration obtains 2-[(2-bromo-4-fluorophenyl) methyl]-1H-isoindole-1,3 (2H)-diketone, be solid (2.06g) that it is not further purified and promptly is used for next step.
LC/MS[M+H]
+=334, retention time=3.30 minute.
(ii) with hydrazine hydrate (0.655ml 21mmol) is added to 2-[(2-bromo-4-fluorophenyl) methyl]-1H-isoindole-1,3 (2H)-diketone (2g, 6mmol) in the suspension in ethanol (60ml), and in stirred overnight at room temperature.At this moment reaction is incomplete, so mixture is in 100 ℃ of heating 2 hours altogether (mixture becomes white and muddy during this period).Filtering mixt removes solid, cooling and filtration then once more.The cold washing with alcohol of solid, the ethanol fraction of vacuum-evaporation merging is to doing then.The gained resistates is dispensed between 2N aqueous hydrochloric acid and the methylene dichloride.Use the hydrophobic glass material to separate organic phase.Water layer is used washed with dichloromethane again, and separates once more.The vacuum concentration water layer produces faint yellow solid (0.876g) then.This solid is dissolved in saturated sodium bicarbonate aqueous solution, and uses dichloromethane extraction.Obtain resistates by separation of hydrophobic glass material and evaporation, it is dissolved in ether and uses the ethereal solution of hydrogenchloride to handle.From mixture, be settled out faint yellow solid.Evaporation and drying obtain [(2-bromo-4-fluorophenyl) methyl] amine hydrochlorate (0.789g).
LC/MS[M+H]
+=203, retention time=1.08 minute.
3) { [3-fluoro-2-(trifluoromethyl) phenyl] methyl } amine hydrochlorate (amine that preparation embodiment 64 is used)
Under the argon gas room temperature, ((1g is in tetrahydrofuran (THF) 4.8mmol) (40ml) solution 19.2mmol) to drop to 3-fluoro-2-(trifluoromethyl) benzamide for 1M, 19.2ml with the borine tetrahydrofuran (THF).Mixture is in 70 ℃ of heating, and then (10ml 10mmol), and heated weekend in 70 ℃ of continuation to add the borine tetrahydrofuran (THF) of another equal portions.Reaction mixture is cooled to room temperature, uses 2M aqueous hydrochloric acid (15ml) to handle then, in stirring at room 15 minutes.Adding aqueous sodium hydroxide solution is 8-9 up to the pH of mixture, and (3 * 30ml) extract mixture with ethyl acetate then.Filter the organic layer that merges, vacuum-evaporation then by the hydrophobic glass material.Resistates is dissolved in the methylene dichloride again, filters, and evaporation obtains yellow oil by the hydrophobic glass material.This oily matter is dissolved in the 2M aqueous hydrochloric acid.Form white precipitate, and collect, then its equivalent is loaded on 4 * 10g SCX post by vacuum filtration.This post uses ammoniacal liquor to wash out product with the flushing of first alcohol and water then.The fraction vacuum concentration of back is obtained yellow oil (0.4g).This oily matter is dissolved in the ether, and handles up to not precipitation generation again with the ethereal solution of 1M hydrogenchloride.The mixture vacuum concentration being obtained { [3-fluoro-2-(trifluoromethyl) phenyl] methyl } amine hydrochlorate is white solid.
LC/MS[M+H]
+=193, retention time=1.15 minute.
Embodiment 65-69
With with embodiment 12 similar methods, prepare the embodiment in the following table (table 5) by replace [(2,3, the 4-trifluorophenyl) methyl] amine that method is used described in the embodiment 12 with suitable amine (or its salt).Unless otherwise indicated, all used amine of compound shown in the preparation table 5 maybe can use the approach described in the chemical literature to prepare from commercially available.Prepare the used 1-ethyl of these embodiment (except the embodiment 65)-5-oxo-proline(Pro) and use the method C described in the embodiment 12, and embodiment 65 using method A.
Table 5
Embodiment 70N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-5-oxo prolineamide (E70)
With 1-ethyl-5-oxo proline(Pro) (0.100g, 0.64mmol) be dissolved in the mixture of methylene dichloride (3ml) and dimethyl formamide (0.5ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.147g, 0.77mmol), I-hydroxybenzotriazole (0.104g, 0.77mmol) and N-ethylmorpholine (0.244ml, 1.92mmol).Stirred the mixture 10 minutes, and then 2-chloro-4-luorobenzyl amine was added to mixture, and continue to stir spend the night (～16 hours) in room temperature.Then mixture with saturated sodium bicarbonate aqueous solution (～2ml) handle and vigorous stirring～10 minute.Use phase splitter to remove water layer, and (2 * 1ml) extract to use methylene dichloride again.Concentrates the organic layer that merges and obtain yellow oil, and it obtains N-[(2-chloro-4-fluorophenyl with the automatic HPLC purifying of mass spectrometric detection subsequently) methyl]-1-ethyl-5-oxo-D-prolineamide (0.065g), be white solid.LC/MS[M+H]
+=299, retention time=2.16 minute.
Enantiomer is excessive=and 80.9%, detect by chiral chromatography B, be indicated as N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-5-oxo-D-prolineamide
Retention time=5.91 minute
1-ethyl-5-oxo proline(Pro) that aforesaid method uses is prepared as follows:
(i) (4.17g 26.53mmol) is dissolved in tetrahydrofuran (THF) (30ml), and (2.23ml 27.86mmol) obtains yellow solution to add iodoethane with D-Pyrrolidonecarboxylic acid ethyl ester.It is cooled to 0 ℃, and the dropping sodium hydride (60% in oil, 1.11g, 27.86mmol).After adding all sodium hydrides, mixture is in 0 ℃ of restir 20 minutes, up to almost no longer producing bubble.Then mixture is warmed to room temperature and under argon gas, stirs and spend the night.Mixture saturated aqueous ammonium chloride (～5ml) processing then.Separate organic layer, and water layer is used methylene dichloride (3 * 20ml) extractions again.The organic layer that merges concentrates then and obtains green/brown oil (3.2g) by the phase splitter drying.It is by automatically quick silica gel column chromatography (Biotage SP4), hexane solution wash-out purifying with 0-100% gradient ethyl acetate, obtain 1-ethyl-5-oxo ethyl prolinate, be yellow oil (1.33g), it is not further purified and just is used for next step.
(ii) (1.33g 7.18mmol) is dissolved in the ethanol (10ml), and is cooled to 0 ℃ in ice bath with 1-ethyl-5-oxo ethyl prolinate.To wherein add the 12.5M aqueous sodium hydroxide solution (1.72ml, 21.53mmol), and in 0 ℃ stir the mixture～4 hours.Vacuum-evaporation ethanol, and aqueous residue is acidified to pH1 with the 2N aqueous hydrochloric acid.The volume of vacuum concentration water is to～3ml, uses the mixture of 3: 1 chloroforms and Virahol then, uses the phase splitter extraction.Concentrate the organic layer that merges and obtain faint yellow oily thing, its vacuum-drying crystallization is obtained 1-ethyl-5-oxo proline(Pro), be white solid (1.12g).
Embodiment 71-82
With with embodiment 70 similar methods, prepare compound in the following table (table 6) by the 2-chloro-4-luorobenzyl amine that replaces with suitable amine (or its salt) using in the aforesaid method.Unless otherwise indicated, all used amine of compound shown in the preparation table 5 maybe can use the approach described in the chemical literature to prepare from commercially available.During detection (by chirality HPLC), the enantiomer of isomer shown in also having listed excessive (e.e.), together with its stereospecificity title, corresponding retention time (r.t.) in used chiral separation method and this method.
Table 6
Embodiment 83N-{[4-fluoro-2-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo prolineamide (E83)
N-{[4-fluoro-2-(trifluoromethyl) phenyl] methyl-1-methyl-5-oxo prolineamide with above-mentioned synthetic N-[(2-chloro-4-fluorophenyl) methyl]-similarly method is synthetic for 1-ethyl-5-oxo prolineamide (embodiment 70), except replacing 1-ethyl-5-oxo proline(Pro) with 1-methyl-5-oxo proline(Pro) (being prepared as follows), and with { [4-fluoro-2-(trifluoromethyl) phenyl] methyl } amine replacement 2-chloro-4-luorobenzyl amine.
LC/MS[M+H]
+=319, retention time=2.14 minute.
1-methyl-5-oxo proline(Pro) that aforesaid method uses is prepared as follows:
(i) (4.0g 25.5mmol) is dissolved in tetrahydrofuran (THF) (25ml), and is cooled to 0 ℃ with D-Pyrrolidonecarboxylic acid ethyl ester.The adding methyl iodide (1.66ml, 26.7mmol), and in 0 ℃ of following continuation stirring of argon gas 10 minutes.(60% in oil, and 1.6g 26.7mmol) (makes every part of reaction) for portioning adding sodium hydride then.After adding all sodium hydrides, under argon gas, make mixture be warmed to room temperature and stir and spend the night.Then mixture with saturated aqueous ammonium chloride (～15ml) handle, and stirred 4 hours.Separate organic layer, and water layer is used dichloromethane extraction again.The organic layer that merges concentrates then and obtains dark oily matter through dried over mgso.By quick silica gel column chromatography, the hexane solution wash-out purifying with 0-75% gradient ethyl acetate obtains 1-methyl-5-oxo ethyl prolinate, is colorless oil (0.27g) that it is not further purified and just is used for next step with it.
(ii) (0.27g 1.58mmol) is dissolved in ethanol (5ml), and is cooled to 0 ℃ in ice bath with 1-methyl-5-oxo ethyl prolinate.To wherein adding 2M aqueous sodium hydroxide solution (3ml), and in 0 ℃ stir the mixture～4 hours.Vacuum-evaporation ethanol, and aqueous residue is acidified to pH1 with the 2N aqueous hydrochloric acid.Volume with water under the vacuum is concentrated into～3ml, uses the mixture of 3: 1 chloroforms and Virahol to use the phase splitter extraction then.Concentrate the organic layer that merges and obtain 1-methyl-5-oxo proline(Pro), it is not further purified promptly and uses.
Embodiment 84-90
And, as similar methods as described in the above-mentioned embodiment 70, prepare the compound of following table (table 7) by replace used 2-chloro-4-luorobenzyl amine among the embodiment 70 with suitable amine (or its salt).Unless otherwise indicated, all used amine of compound shown in the preparation table 7 maybe can use the approach described in the chemical literature to prepare from commercially available.Replace used 1-ethyl-5-oxo proline(Pro) among the embodiment 70 with 1-methyl-5-oxo proline(Pro) (as above-mentioned embodiment 81 preparations).When detecting (by chirality HPLC), the enantiomer of isomer shown in having listed excessive (e.e.), with and the stereospecificity title, corresponding retention time (r.t.) in used chiral separation method and this method.
Table 7
Embodiment 91N-[(2-chloro-4-fluorophenyl) methyl]-5-oxo-1-phenyl-prolineamide (E91)
With 5-oxo-1-phenyl-proline(Pro) (0.072g, 0.35mmol, be prepared as follows) be dissolved in methylene dichloride (～2ml) and in the dimethyl formamide (0.5ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.081g, 0.42mmol), I-hydroxybenzotriazole (0.057g, 0.42mmol) and N-ethylmorpholine (0.134ml, 1.05mmol).Mixture is in stirring at room 30 minutes, add then [(2-chloro-4-fluorophenyl) methyl] amine (0.067g, 0.42mmol).Continue to stir in room temperature, mixture is again with methylene dichloride and saturated sodium bicarbonate aqueous solution dilution then.Separate water layer, and use methylene dichloride (3 equal portions) extraction again.The organic layer of He Binging salt water washing then is then through dried over mgso.Evaporating solvent obtains yellow oil then, and it is by the automatic HPLC purifying of mass spectrometric detection.The material that will obtain thus grinds with the mixture of 1: 1 methylene dichloride and ether at last, filter and drying after obtain pure N-[(2-chloro-4-fluorophenyl) methyl]-5-oxo-1-phenyl-prolineamide (0.031g), be white solid.LC/MS[M+H]
+=347, retention time=2.46 minute.
5-oxo-1-phenyl-the proline(Pro) that uses in the aforesaid method is prepared as follows:
(i) with D-Pyrrolidonecarboxylic acid ethyl ester (0.200g, 1.27mmol) be dissolved in the diox (5ml), and with three (dibenzalacetones) close two palladiums (0) (0.058g, 0.06mmol), bromobenzene (0.351ml, 1.53mmol), cesium carbonate (0.621g, 1.91mmol) and Xantphos
TM(0.110g 0.19mmol) handles.The gained mixture spends the night in reflux, is cooled to room temperature then.Mixture dilutes with methyl alcohol, and filters.Vacuum-evaporation filtrate is allocated between methylene dichloride and the saturated sodium bicarbonate aqueous solution then.Water layer is used methylene dichloride (3 equal portions) extraction again, and the organic layer of He Binging salt water washing then is then through dried over mgso.Evaporating solvent obtains the glassy yellow resistates, and it is by quick silica gel column chromatography, and the hexane solution wash-out purifying with 0-50% gradient ethyl acetate obtains 5-oxo-1-phenyl proline methyl ester (0.078g), is yellow oil.It is not further purified and just is used for next step.
(ii) (0.078g 0.36mmol) merges in ethanol (2ml) with 2N aqueous sodium hydroxide solution (2ml) with 5-oxo-1-phenyl proline methyl ester in 0 ℃.Stirred the mixture 5 hours in-10 ℃ to 0 ℃.Vacuum evaporating solvent then, resistates is acidified to pH1 by adding the 2M aqueous hydrochloric acid.To wherein adding methylene dichloride, and make mixture pass through phase splitter.Water layer is used washed with dichloromethane again, and the dichloromethane layer of evaporation merging obtains 5-oxo-1-phenyl-proline(Pro) (0.072g) then, is yellow jelly, and it is not further purified and promptly is used for next step.
Embodiment 92N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-5-oxo-1-(phenyl methyl)-prolineamide (E92)
With 5-oxo-1-(phenyl methyl) proline(Pro) (0.100g, 0.46mmol, be prepared as follows) be dissolved in the mixture of methylene dichloride (2.5ml) and dimethyl formamide (0.5ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.105g, 0.55mmol), I-hydroxybenzotriazole (0.074g, 0.55mmol) and N-ethylmorpholine (0.143ml, 1.37mmol).Mixture was stirred 10 minutes, and (0.115g's amine 0.55mmol), and stirred the mixture 1 hour to add { [2-chloro-3-(trifluoromethyl) phenyl] methyl } then.Add saturated sodium bicarbonate aqueous solution (10ml), and vigorous stirring mixture 15 minutes.Organic phase is separated with phase splitter, and water is used methylene dichloride (3 * 10ml) washings of several equal portions again.Merge organic fraction, and through dried over mgso.Evaporating solvent, and resistates then obtains pure N-{[2-chloro-3-(trifluoromethyl) phenyl by the automatic HPLC purifying of mass spectrometric detection] methyl }-5-oxo-1-(phenyl methyl)-D-prolineamide.
LC/MS[M+H]
+=411, retention time=2.77 minute.
Enantiomer is excessive=and 100.0%, detect by chiral chromatography D, be indicated as N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-5-oxo-1-(phenyl methyl)-D-prolineamide.
Retention time=10.58 minute
5-oxo-1-(phenyl methyl) proline(Pro) that uses in the aforesaid method is prepared as follows:
With D-L-glutamic acid (1.47g, 10mmol) be dissolved in the 2N aqueous sodium hydroxide solution (10ml, 20mmol) in, and stirred 15 minutes.Mixture is with phenyl aldehyde (1.1ml, ethanol 10mmol) (3ml) solution-treated, and in stirring at room 30 minutes then.Mixture is cooled to 0 ℃, and handles with sodium borohydride (0.030g).Make mixture be warmed to room temperature through 4 hours and stir simultaneously, use ether (3 times) washing then, be acidified to pH2 with concentrated hydrochloric acid then.Leach the gained precipitation, and wash with ether, pulp in ethanol then, and use the ethanol azeotropic again 3 times.Make residual substance pulp in ethanol (50ml) at last, and reflux 16 hours.Then mixture is cooled to room temperature, and vacuum-evaporation.Drying obtains pure 5-oxo-1-(phenyl methyl) proline(Pro).
Embodiment 93N-[(2-chloro-4-fluorophenyl) methyl]-5-oxo-1-(phenyl methyl) prolineamide (E93)
With with above-mentioned synthetic N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl-5-oxo-1-(phenyl methyl) prolineamide (embodiment 92) similar methods prepares N-[(2-chloro-4-fluorophenyl) methyl]-5-oxo-1-(phenyl methyl) prolineamide, replace { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine except using [(2-chloro-4-fluorophenyl) methyl] amine.
LC/MS[M+H]
+=361, retention time=2.54 minute.
Embodiment 94N-[(2-chloro-4-fluorophenyl) methyl]-1-cyclopentyl-5-oxo prolineamide (E94)
With 1-cyclopentyl-5-oxo proline(Pro) (0.100g, 0.51mmol, be prepared as follows) be dissolved in the mixture of methylene dichloride (2.5ml) and dimethyl formamide (0.5ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.117g, 0.61mmol), I-hydroxybenzotriazole (0.082g, 0.61mmol) and N-ethylmorpholine (0.2ml, 1.52mmol).Stirred the mixture 10 minutes, (0.097g 0.61mmol), and stirs the mixture and spends the night to add [(2-chloro-4-fluorophenyl) methyl] amine then.Add saturated sodium bicarbonate aqueous solution (10ml), and vigorous stirring mixture 15 minutes.Organic phase is separated with phase splitter, and water is used methylene dichloride (3 * 10ml) washings of several equal portions again.Merge organic fraction, and through dried over mgso.Evaporating solvent, and resistates then obtains pure N-[(2-chloro-4-fluorophenyl by the automatic HPLC purifying of mass spectrometric detection) methyl]-1-cyclopentyl-5-oxo prolineamide.
LC/MS[M+H]
+=339, retention time=2.4 minute.
The 1-cyclopentyl of using in the aforesaid method-5-oxo proline(Pro) is prepared as follows:
(i) with D-glutamic acid dimethyl ester hydrochloride (2.1g 10.00mmol) is dissolved in methyl alcohol (7.5ml) and the tetrahydrofuran (THF) (15ml), and under the argon gas mixture (0.402g 10.05mmol) handled 20 minutes with the sodium hydroxide of pulverizing.At this moment, with acetate (0.575ml, 10.05mmol) and cyclopentanone (0.889ml 10.05mmol) is added in the mixture.Stir after 10-15 minute, mixture is cooled to 0 ℃ in ice bath, and (0.380g 10.05mmol) handles with the sodium borohydride sheet.Stirred the mixture under the argon gas 3 hours, and be warmed to room temperature.When mixture reaches room temperature, evaporate methyl alcohol, and resistates is with methylene dichloride (20ml) dilution, and with saturated sodium bicarbonate aqueous solution (～25ml) wash.Separate organic layer, and water layer is used methylene dichloride again, and (2 * 20ml) strip.Concentrate the organic layer that merges in the vacuum and obtain oily matter.Oily matter is dissolved in the toluene (10ml), and reflux spends the night.The resistates of evaporating solvent, and gained then is by quick silica gel column chromatography, obtains thick 1-cyclopentyl-5-oxo proline methyl ester with the dichloromethane solution wash-out purifying of 0-10% gradient methyl alcohol, and it is not further purified and promptly is used for next step.
(ii) (0.560g 2.65mmol) is dissolved in the ethanol (10ml), and is cooled to 0 ℃ in ice bath with 1-cyclopentyl-5-oxo proline methyl ester.Add 2M aqueous sodium hydroxide solution (5ml), and stirred the mixture 4 hours in ice temperature.Vacuum-evaporation ethanol, and aqueous residue then is acidified to pH1 by adding the 2N aqueous hydrochloric acid.The volume of vacuum concentration gained aqueous mixture is to～3ml, uses the mixture of 3: 1 chloroforms and Virahol to use the phase splitter extraction then.Water layer is used washed with dichloromethane again, and organic fraction of evaporation merging obtains thick 1-cyclopentyl-5-oxo proline(Pro) then, and it is not further purified and just is used for subsequent reaction.
Embodiment 95-99
With with similar methods described in the embodiment 94, by with [(the 2-chloro-4-fluorophenyl) methyl] amine that uses in suitable amine (or its salt) the replacement aforesaid method and/or with the cyclopentanone that uses in the suitable aldehydes or ketones replacement aforesaid method, prepare the compound in the following table (table 8).Unless otherwise indicated, all amine of the use of compound shown in the preparation table 8 prepare from the commercially available described approach of chemical literature that maybe can use.
Table 8
Embodiment 100N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-(2, the 2-dimethyl propyl)-5-oxo prolineamide (E100)
With 1-(2, the 2-dimethyl propyl)-5-oxo proline(Pro) (0.100g, 0.5mmol, be prepared as follows) be dissolved in the methylene dichloride (5ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.191g, 1mmol) and I-hydroxybenzotriazole (0.135g, 1mmol).In stirring at room mixture 30 minutes, (0.209g, 1mmol), and mixture at room temperature stirred and spends the night to add { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine then.Mixture water successively then, the washing of 3N aqueous citric acid solution, and wash with water again 3 times, then by hydromatrix post (Varian 5g) filtration drying.Evaporating solvent, and resistates then obtains pure N-{[2-chloro-3-(trifluoromethyl) phenyl by the automatic HPLC purifying of mass spectrometric detection] methyl }-1-(2, the 2-dimethyl propyl)-5-oxo prolineamide.
LC/MS[M+H]
+=391/393, retention time=2.78 minute.
The 1-that uses in the aforesaid method (2, the 2-dimethyl propyl)-5-oxo proline(Pro) is prepared as follows:
With L-L-glutamic acid (1.47g, 10mmol) be dissolved in the 2N aqueous sodium hydroxide solution (10ml, 20mmol) in, and with trimethyl-acetaldehyde (1.09ml, ethanol 10mmol) (5ml) solution-treated is then in stirring at room 30 minutes.Mixture is cooled to 0 ℃, and handles with sodium borohydride (0.130g).Through 4 hours mixture is warmed to room temperature, stirs simultaneously, be acidified to neutral pH then.Pulp in ethanol behind the vacuum concentration, and use the ethanol azeotropic again 3 times.At last, residual substance is suspended in the ethanol (50ml), and in reflux 48 hours.Then mixture is cooled to room temperature, leaches salt, and vacuum evaporating solvent obtains jelly.Grind dry then pure solid 1-(2, the 2-the dimethyl propyl)-5-oxo proline(Pro) (1.1g) that obtains with ether.
Embodiment 101N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-5-oxo-1-(phenyl methyl)-prolineamide (E101)
With above-mentioned synthetic N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-(2, the 2-dimethyl propyl)-5-oxo prolineamide (embodiment 100) similar methods prepares N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-5-oxo-1-(phenyl methyl)-D-prolineamide, except using 5-oxo-1-(phenyl methyl) proline(Pro) to replace 1-(2, the 2-dimethyl propyl)-5-oxo proline(Pro).
LC/MS[M+H]
+=411/413, retention time=2.77 minute.
Enantiomer is excessive=and 100.0%, detect by chiral chromatography D, be indicated as N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-5-oxo-1-(phenyl methyl)-D-prolineamide.
Retention time=8.09 minute.
Prepare 5-oxo-1-(phenyl methyl) proline(Pro) with above-mentioned synthetic 1-(2, the 2-dimethyl propyl)-described similar method of 5-oxo proline(Pro) (embodiment 100), replace trimethyl-acetaldehyde except using phenyl aldehyde.
Embodiment 102N-[(2, the 4-dichlorophenyl) methyl]-1-methyl-5-oxo prolineamide (E102)
With above-mentioned synthetic N-{[4-fluoro-2-(trifluoromethyl) phenyl] methyl }-1-methyl-described similar methods of 5-oxo prolineamide (embodiment 83) prepares N-[(2, the 4-dichlorophenyl) methyl]-1-methyl-5-oxo prolineamide, except replacing { [4-fluoro-2-(trifluoromethyl) phenyl] methyl } amine with [(2,4 dichloro benzene base) methyl] amine.
LC/MS[M+H]
+=300.9, retention time=2.13 minute.
Enantiomer is excessive=and 97.8%, detect by chiral chromatography A, be indicated as N-[(2, the 4-dichlorophenyl) methyl]-1-methyl-5-oxo-D-prolineamide.
Retention time=6.25 minute.
Embodiment 1031-ethyl-N-{[2-fluoro-3-(trifluoromethyl) phenyl] methyl }-5-oxo prolineamide (E103)
With above-mentioned synthetic N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-described similar methods of 5-oxo-D-prolineamide (seeing embodiment 70) prepares 1-ethyl-N-{[2-fluoro-3-(trifluoromethyl) phenyl] methyl }-5-oxo prolineamide, replace 2-chloro-4-luorobenzyl amine except using 2-fluoro-3-trifluoromethyl benzyl amine.
LC/MS[M+H]
+=333, retention time=2.24 minute.
Embodiment 104-109
With with embodiment 12 described similar methods, by replace used [(2,3, the 4-trifluorophenyl) methyl] amine in the foregoing description 12, the embodiment of preparation following table (table 9) with suitable amine (or its salt).Unless otherwise indicated, all amine of the use of compound shown in the preparation table 9 maybe can use the approach described in the chemical literature to prepare from commercially available.Prepare the used 1-ethyl of these embodiment-5-oxo-proline(Pro) and use for example preparation of the method C described in 12.
Table 9
2-(amino methyl)-6-(trifluoromethyl) cyanobenzene trifluoroacetate that synthetic embodiment 105 needs is prepared as follows:
(i) (1.93g 10mmol) is dissolved in methylene dichloride (40ml), and with two (1, the 1-dimethyl ethyl) two carbonic ethers (2.18g, methylene dichloride 10mmol) (10ml) solution-treated with { [2-fluoro-3-(trifluoromethyl) phenyl] methyl } amine.In stirring at room after 2 hours, evaporating solvent obtains faint yellow solid, and it is by silica gel column chromatography, with 1: 10-1: the hexane solution wash-out purifying of 5 gradient ethyl acetate, obtain pure { [2-fluoro-3-(trifluoromethyl) phenyl] methyl } carboxylamine 1,1-dimethyl ethyl ester (2g).
(ii) with { [2-fluoro-3-(trifluoromethyl) phenyl] methyl } carboxylamine 1, (1.17g 4mmol) is dissolved in the methyl-sulphoxide (5ml) 1-dimethyl ethyl ester, and (0.260g 4mmol) handles with potassium cyanide.Mixture heated 1.5 hours down in 80 ℃ of argon gas then, then in 120 ℃ of heated overnight (16 hours).And then (0.260g 4mmol), and continues heating 24 hours again in 120 ℃ to add potassium cyanide.Then mixture is cooled to room temperature, the water cancellation, and dilute with ethyl acetate.Separate organic extract liquid, and wash with water 3 times, wash with saturated sodium-chloride water solution then.Dry and evaporation obtains the brown jelly, it passes through silica gel column chromatography, with 1: 10-1: the hexane solution wash-out purifying of 5 gradient ethyl acetate, obtain partial-purified { [2-cyano group-3-(trifluoromethyl) phenyl] methyl } carboxylamine 1,1-dimethyl ethyl ester, be dark solid/semisolid, it is not further purified and just is used for next step.
LC/MS[M-BOC+H]
+=201, retention time=1.19 minute.
(iii) with { [2-cyano group-3-(trifluoromethyl) phenyl] methyl } carboxylamine 1, (0.190g 0.63mmol) is dissolved in methylene dichloride (4ml) to 1-dimethyl ethyl ester, and handles with trifluoroacetic acid (4ml).Mixture is in stirring at room 1 hour, evaporation then.Resistates is dissolved in methylene dichloride 2 times, and evaporation obtains thick 2-(amino methyl)-6-(trifluoromethyl) cyanobenzene trifluoroacetate once more, and it is not further purified promptly and uses.
Embodiment 1101-methyl-N-(1-naphthyl methyl)-5-oxo prolineamide (E110)
With 1-methyl-5-oxo proline(Pro) (0.050g, 0.35mmol, with the described similar methods preparation of the foregoing description 51), N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.081g, 0.42mmol), I-hydroxybenzotriazole (0.057g, 0.42mmol), N-ethylmorpholine (0.166ml, 1.05mmol) and (1-naphthyl methyl) amine be incorporated in methylene dichloride (～8ml) in, and mixture is in stirring at room～20 hour.Mixture washs with 2M aqueous hydrochloric acid (5ml) then, and uses phase splitter to separate organic layer.Organic layer washs with saturated sodium bicarbonate aqueous solution, as preceding separation, and evaporation then.Resistates obtains pure 1-methyl-N-(1-naphthyl methyl)-5-oxo prolineamide by the automatic HPLC purifying of mass spectrometric detection, is white solid (0.062g).
LC/MS[M+H]
+=283, retention time=2.1 minute.
Embodiment 111N-{[2-chloro-4-fluoro-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo prolineamide (E111)
(0.057g, 0.4mmol is with the described similar methods preparation of the foregoing description 51) is dissolved in the methylene dichloride (4ml) with 1-methyl-5-oxo proline(Pro), and with 2-oxyethyl group-1-ethoxy carbonyl-1,2-dihydroquinoline (0.104g, 0.42mmol) processing.Add { [2-chloro-4-fluoro-3-(trifluoromethyl) phenyl] methyl } amine hydrochlorate (0.105g, 0.4mmol are prepared as follows) then, and mixture was in stirring at room 4 hours.Mixture is handled with saturated sodium bicarbonate aqueous solution (10ml), and stirs 5 minutes.Use the hydrophobic glass material to separate organic phase, use (2 * 10ml) washings of 2N aqueous hydrochloric acid then.Evaporate organic phase then and obtain pure N-{[2-chloro-4-fluoro-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo prolineamide (0.106g).
LC/MS[M+H]
+=353, retention time=2.49 minute.
{ [2-chloro-4-fluoro-3-(trifluoromethyl) phenyl] methyl } amine hydrochlorate that aforesaid method uses is prepared as follows:
(i) (10g 50mmol) is dissolved in the tetrahydrofuran (THF) (100ml), is cooled to-70 ℃ under the argon gas, and with hexanaphthene (37.5ml, 52.5mmol) solution-treated of 1.4M s-butyl lithium with 1-chloro-3-fluoro-2-(trifluoromethyl) benzene.Continue to stir 2 hours, (6.7ml 52.5mmol), and still continues to stir 1 hour in-70 ℃ again to add trimethylsilyl chloride then.Mixture is warmed to room temperature, and vacuum removes tetrahydrofuran (THF) then.Resistates is allocated between ether and the water, separates organic layer then, and wash with the 2N aqueous hydrochloric acid.Separate organic phase, and the concentrated crude product that obtains, it obtains pure [4-chloro-2-fluoro-3-(trifluoromethyl) phenyl] (trimethylammonium) silane by quick silica gel column chromatography with hexane wash-out purifying, is clarification oily matter (10.35g).
(ii) under-75 ℃ of argon gas, with 2,2,6, (3.3ml, (2.5M is in toluene, and 7.7ml in tetrahydrofuran (THF) 19.44mmol) (75ml) solution, and stirred 15 minutes 19.44mmol) slowly to be added to n-Butyl Lithium for the 6-tetramethyl piperidine.(5g, tetrahydrofuran (THF) 18.5mmol) (10ml) drips of solution adds in the mixture, guarantees that mixture temperature keeps below-65 ℃, and continues to stir 2 hours with [4-chloro-2-fluoro-3-(trifluoromethyl) phenyl] (trimethylammonium) silane then.Add block excessive solidified carbon dioxide (washing with tetrahydrofuran (THF) prior to-65 ℃ in advance), and mixture was warmed to room temperature through 2 hours.Mixture obtains faint yellow solid in vacuum concentration.With this dispensed materials between water (it is acidified to pH1 (200ml)) and ether (200ml).Separate organic layer, and through anhydrous sodium sulfate drying.Evaporation obtains filbert solid, and it obtains pure 2-chloro-4-fluoro-3-(trifluoromethyl)-5-(trimethyl silyl) phenylformic acid (3.85g divides 3 batches) by the toluene recrystallization, is white needles.
LC/MS[M-H]
-=312, retention time=3.29 minute.
(iii) with Potassium monofluoride (0.367g, 9.55mmol) water (15ml) solution be added to 2-chloro-4-fluoro-3-(trifluoromethyl)-5-(trimethyl silyl) phenylformic acid (1g, 3.18mmol) tetrahydrofuran (THF) (50ml) solution in, and mixture spends the night in 100 ℃ of stirrings.(0.370g 9.62mmol), and continues heating 4 hours in 100 ℃ to add another part water (15ml) and Potassium monofluoride.The vacuum-evaporation tetrahydrofuran (THF), and replace with the dissolving all solids with the capacity dimethyl formamide.Mixture is in 100 ℃ of heated overnight, but still has starting material, and (0.367g 9.55mmol), and continues heating 7 days in 100 ℃ therefore to add Potassium monofluoride again.At this moment, nearly all initial substance disappears, and evaporation reaction is extremely done under the vacuum, and is dissolved in 2N aqueous hydrochloric acid (75ml) and the ether (50ml).Separate water layer, and (2 * 50ml) extractions, the organic grade of lease making dried over sodium sulfate that will merge then, and evaporate and obtain crude product are white solid to use ether again.It is white solid by obtained pure 2-chloro-4-fluoro-3-(trifluoromethyl) phenylformic acid (0.566g) by recrystallization purifying in the toluene.
(iv) with 2-chloro-4-fluoro-3-(trifluoromethyl) phenylformic acid (0.560g, 2.31mmol), 1H-1,2,3-benzotriazole-1-acid ammonium (ammonium 1H-1,2,3-benzotriazol-1-olate) (0.534g, 3.47mmol, be prepared as follows), N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.643g, 3.47mmol) and N-ethylmorpholine (0.594ml 4.62mmol) stirred in methylene dichloride (30ml) 3 hours together totally.Add saturated sodium bicarbonate aqueous solution (30ml), and stirred the mixture 15 minutes.Use the hydrophobic glass material to separate organic layer, use (2 * 50ml) washings of 2N aqueous hydrochloric acid then.Reuse the hydrophobic glass material and separate organic layer, and vacuum-evaporation obtains 2-chloro-4-fluoro-3-(trifluoromethyl) benzamide (0.493g), be pale solid, it is not further purified and promptly is used for next step.
The 1H-1 that uses in the above-mentioned steps, 2,3-benzotriazole-1-acid ammonium is prepared as follows:
In 0 ℃ (ice bath), (4.15ml, (10g in tetrahydrofuran (THF) 74mmol) (100ml) solution, and stirred 2 hours 75mmol) slowly to be added to I-hydroxybenzotriazole with ammonium hydroxide.Filter and obtain 1H-1 with the tetrahydrofuran (THF) washing, 2,3-benzotriazole-1-acid ammonium (10.57g) is white solid.
(v) (0.490g is 2.03mmol) with the tetrahydrofuran (THF) of 1M borine (20.33ml, 20.33mmol) solution-treated, and spend the night in 60 ℃ of stirrings for 2-chloro-4-fluoro-3-(trifluoromethyl) benzamide.Mixture is handled up to the gas generation with the 2N aqueous hydrochloric acid and is stopped then, stirs 2 hours in 100 ℃ then.Vacuum concentrated mixture, and resistates is dissolved in the less water, and with washed with dichloromethane (30ml).Regulate the pH to pH 11 of water layer by adding the 2N aqueous sodium hydroxide solution, use methylene dichloride (2 * 25ml) extractions then.Use hydrophobic glass material separate dichloromethane layer, merging also, vacuum-evaporation obtains faint yellow oily thing.(it is not further purified promptly and uses for 3ml, 3mmol) solution, and leach the gained white solid and obtain pure { [2-chloro-4-fluoro-3-(trifluoromethyl) phenyl] methyl } amine hydrochlorate (0.210g) to add the ether of 1M hydrogenchloride.
Embodiment 112N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-cyclobutyl-5-oxo prolineamide (E112)
With 1-cyclobutyl-5-oxo proline(Pro) (0.238g, 0.82mmol) be suspended in the methylene dichloride (3ml), and to wherein adding N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.188g, 0.98mmol), I-hydroxybenzotriazole (0.132g, 0.98mmol) and N-ethylmorpholine (0.313ml, 2.46mmol).Mixture was in stirring at room 30 minutes, and (0.205g 0.98mmol) is added in the mixture, and continues to stir～20 hours in room temperature with { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine then.Mixture dilutes with methylene dichloride more then, and handles with saturated sodium bicarbonate aqueous solution.The separate dichloromethane layer, and water layer is used 3 equal portions dichloromethane extractions again.The organic extract liquid that merges washes with water, uses the salt water washing then, and through anhydrous magnesium sulfate drying, and vacuum-evaporation obtains crude product.It further obtains pure N-{[2-chloro-3-(trifluoromethyl) phenyl by the automatic HPLC purifying of mass spectrometric detection] methyl }-1-cyclobutyl-5-oxo prolineamide (0.105g), be white solid.
LC/MS[M+H]
+=375, retention time=2.53 minute.
The 1-cyclobutyl of using in the aforesaid method-5-oxo proline(Pro) is to prepare with the described similar methods of synthetic 1-ethyl-5-oxo-proline methyl ester (referring to embodiment 3); except using cyclobutanone to replace acetaldehyde; and carry out subsequently ester deprotection steps (use standard conditions, the i.e. methanol solution of sodium hydroxide) (opposite with the acid amides coupling) with the deprotection of the merging described in the embodiment 3.
Embodiment 113-117
With with similar methods described in the foregoing description 112, used { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine prepares the compound in the following table (table 10) in the aforesaid method by replacing with suitable amine (or its salt).Unless otherwise indicated, all used amine of compound shown in the preparation table 10 maybe can use the approach described in the chemical literature to prepare from commercially available.
Table 10
Synthetic N-[(2-chloro-3, the 4-difluorophenyl) methyl]-required as above embodiment 36 preparations of [(2-chloro-3,4-difluorophenyl) methyl] amine hydrochlorate of 1-cyclobutyl-5-oxo prolineamide (embodiment 115).
Embodiment 118N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-(2, the 2-dimethyl propyl)-5-oxo prolineamide (E118)
To prepare N-{[2-chloro-3-(trifluoromethyl) phenyl with similar methods described in the embodiment 100] methyl }-1-(2, the 2-dimethyl propyl)-5-oxo prolineamide, except using 1-(2, the 2-dimethyl propyl)-5-oxo proline(Pro) (being prepared as follows).
LC/MS[M+H]
+=391/393, retention time=2.76 minute.
1-(2, the 2-dimethyl propyl)-5-oxo proline(Pro) that aforesaid method uses is prepared as follows:
With D-L-glutamic acid (2.21g, 15mmol) be dissolved in the 2N aqueous sodium hydroxide solution (15ml, 30mmol) in, be cooled to 0 ℃, and (1.63ml, ethanol 15mmol) (3ml) solution-treated is then in stirring at room 45 minutes with trimethyl-acetaldehyde.Mixture is cooled to 0 ℃ once more, and use sodium borohydride (0.189g 5mmol) handles in batches.Make mixture be warmed to room temperature through 4 hours, stir simultaneously, with after the ether washing, use concentrated hydrochloric acid to be acidified to～pH4 then.Collect the gained precipitation by filtering, with ether washing, dried overnight in vacuum drying oven then.Solid is suspended in the ethanol (50ml) then, and mixture is in the heating down 24 hours that refluxes.Concentrate then and obtain 1-(2, the 2-dimethyl propyl)-5-oxo proline(Pro) (1.51g) with the hexane grinding, be solid, it is not further purified promptly and uses.
Embodiment 119N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-5-oxo-1-(2-pyridylmethyl) prolineamide (E119)
In room temperature, with 5-oxo-1-(2-pyridylmethyl) proline(Pro) (0.220g, 1mmol, be prepared as follows), N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.384g, 2mmol) and I-hydroxybenzotriazole (0.308g 2mmol) stirred in methylene dichloride (10ml) 30 minutes together.(0.314g, at room temperature stir and spend the night by 1.5mmol) processing, and mixture with { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine for mixture then.Enriched mixture, and be allocated between ethyl acetate and the saturated sodium bicarbonate aqueous solution.Separate water layer, and use ethyl acetate extraction, the ethyl acetate fraction of He Binging is washed with saturated sodium-chloride water solution then with 3 parts of water washings then.Through dried over sodium sulfate, and concentrate and to obtain solid residue, its automatic HPLC purifying by mass spectrometric detection obtains N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-5-oxo-1-(2-pyridylmethyl) prolineamide (0.263g), be beige solid.
LC/MS[M+H]
+=412/414, retention time=2.15 minute.
5-oxo-the 1-that uses in the aforesaid method (2-pyridylmethyl) proline(Pro) is prepared as follows:
In 0 ℃ with D-L-glutamic acid (2.21g, 15mmol) be dissolved in the 2N aqueous sodium hydroxide solution (15ml, 30mmol) in, (1.43ml 15mmol) handles to use pyridine-2-formaldehyde then.Mixture is cooled to 0 ℃ then in stirring at room 45 minutes, and (0.189g 5mmol) handles with sodium borohydride.Made mixture be warmed to room temperature through 4 hours, stir simultaneously, use ether washing 2 times then after, be acidified to pH5-6.Concentrate water layer, use methylbenzene azeotropic then 3 times, use 1: 1 ethanol then: the toluene mixture azeotropic, use the ethanol azeotropic at last.Resistates is dissolved in ethanol (50ml) then, and refluxes 8 hours.Concentrate and obtain oily matter, vacuum-drying obtains 5-oxo-1-(2-pyridylmethyl) proline(Pro) (2.60g) then, is spumescence, and it promptly uses without any further purifying.
Embodiment 120N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-5-oxo-1-(3-pyridylmethyl) prolineamide (E120)
In room temperature with 5-oxo-1-(3-pyridylmethyl) proline(Pro) (0.210g, 1mmol, be prepared as follows), N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.383g, 2mmol) and I-hydroxybenzotriazole (0.306g 2mmol) stirred in methylene dichloride (10ml) 30 minutes together.(0.314g, at room temperature stir and spend the night by 1.5mmol) processing, and mixture with { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine for mixture then.Enriched mixture, and be allocated between ethyl acetate and the saturated sodium bicarbonate aqueous solution.Separate water layer, and use ethyl acetate extraction again, the ethyl acetate fraction of He Binging is by washing with saturated sodium-chloride water solution then with 3 parts of water successively then.Through dried over mgso, and concentrate and to obtain solid residue, its automatic HPLC purifying by mass spectrometric detection obtains pure N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-5-oxo-1-(3-pyridylmethyl) prolineamide (0.031g).
LC/MS[M+H]
+=412/414, retention time=1.83 minute.
5-oxo-the 1-that uses in the aforesaid method (3-pyridylmethyl) proline(Pro) is prepared as follows:
In 0 ℃ with D-L-glutamic acid (2.21g, 15mmol) be dissolved in the 2N aqueous sodium hydroxide solution (15ml, 30mmol) in, use pyridine-3-formaldehyde (1.41ml, ethanol 15mmol) (3ml) solution-treated then.Mixture is cooled to 0 ℃ then in stirring at room 30 minutes, and with sodium borohydride (0.189g, 5mmol) batch treatment.Make mixture be warmed to room temperature through 4 hours, stir simultaneously, with after the ether washing, use concentrated hydrochloric acid to be acidified to pH5-6 then.Collect the gained precipitation by filtering, with the ether washing, and vacuum-drying.Then the gained material is dissolved in ethanol (50ml), and refluxes and spend the night.Remove fine-grained solids by filtration, concentrate then and obtain 5-oxo-1-(3-pyridylmethyl) proline(Pro) (2.04g), be white solid, it promptly uses without any further purifying.
Embodiment 121N-[(2, the 4-dichlorophenyl) methyl]-5-oxo-1-(3-pyridylmethyl) prolineamide (E121)
With with above-mentioned synthetic N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl-the described similar methods of 5-oxo-1-(3-pyridylmethyl) prolineamide (E120) prepares N-[(2, the 4-dichlorophenyl) methyl]-5-oxo-1-(3-pyridylmethyl) prolineamide, except using [(2,4 dichloro benzene base) methyl] amine to replace { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine.
LC/MS[M+H]
+=378/380/382, retention time=1.70 minute.
Embodiment 1221-cyclopropyl-N-[(2, the 4-dichlorophenyl) methyl]-2-methyl-5-oxo prolineamide (E122)
To (2,4 dichloro benzene base) methyl isocyanide (0.047g, 0.25mmol) and levulinic acid (0.041ml, add in methyl alcohol 0.4mmol) (2ml) solution cyclopropylamine (0.042ml, 0.6mmol).Mixture is heated to 100 ℃ in microwave reactor, kept 30 minutes.Vacuum removes solvent, and the automatic HPLC purifying of resistates by mass spectrometric detection obtains 1-cyclopropyl-N-[(2, the 4-dichlorophenyl) methyl]-2-methyl-5-oxo prolineamide (0.054g), be white solid.
LC/MS[M+H]
+=341/343, retention time=2.57 minute.
Embodiment 123-126
With with the foregoing description 122 described similar methods, prepare compound in the following table (table 11) by the cyclopropylamine that replaces with suitable amine using in the aforesaid method.All used amine of compound shown in the preparation table 11 maybe can use the approach described in the chemical literature to prepare from commercially available.
Table 11
Embodiment 127N-[(2, the 4-dichlorophenyl) methyl]-1,3,3-trimethylammonium-5-oxo prolineamide (E127)
(0.094g 0.5mmol) with 3, adds methylamine (0.080ml, 33% ethanolic soln) in methyl alcohol (2ml) solution of 3-dimethyl-4-ketobutyric acid (0.065mg, 0.5mmol are prepared as follows) to (2,4 dichloro benzene base) methyl isocyanide.Mixture is heated to 100 ℃ in microwave reactor, kept 30 minutes.Vacuum removes solvent, and the automatic HPLC purifying of resistates by mass spectrometric detection obtain colourless jelly, and it grinds with ether and obtains N-[(2,4-dichlorophenyl) methyl]-1,3,3-trimethylammonium-5-oxo prolineamide (0.043g) is the thickness white solid.
LC/MS[M+H]
+=329/331, retention time=2.42 minute.
Used 3 in the aforesaid method, 3-dimethyl-4-ketobutyric acid is prepared as follows:
With 3, (1.3g 10.14mmol) is dissolved in methylene dichloride (25ml) to 3-dimethyl-4-pentenoic acid, and is cooled to-78 ℃ in the CO2/ acetone bath.Blast oxygen and passed through mixture 5 minutes, blast ozone 25 minutes (obtaining blue solution) then.Blast oxygen again and passed through mixture 5 minutes, blasted argon gas then 10 minutes.(2.23ml 30.4mmol) is added in the mixture, and shifts out mixture from cooling bath, and stirs 2.5 hours with dimethyl sulfoxide (DMSO) then.Vacuum concentration gained colourless solution obtains 3, and 3-dimethyl-4-ketobutyric acid is colorless oil, and it is not further purified promptly and uses.
Embodiment 128N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1,3,3-trimethylammonium-5-oxo prolineamide (E128)
With with above-mentioned synthetic N-[(2, the 4-dichlorophenyl) methyl]-1,3,3-trimethylammonium-5-oxo prolineamide (E127) similar methods prepares N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1,3,3-trimethylammonium-5-oxo prolineamide replaces (2,4 dichloro benzene base) methyl isocyanide except using [2-chloro-3-(trifluoromethyl) phenyl] methyl isocyanide (as described in example 40 above).
LC/MS[M+H]
+=363/365, retention time=2.49 minute.
Embodiment 129N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1,3-dimethyl-5-oxo prolineamide (E129)
With 1,3-dimethyl-5-oxo proline(Pro) (0.620g, 3.6mmol, be prepared as follows), N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.822g, 4.3mmol), I-hydroxybenzotriazole (0.581g, 4.3mmol), N-ethylmorpholine (1.4ml, 10.8mmol) and { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine (0.828g, 3.96mmol) merge in the mixture of methylene dichloride (10ml) and dimethyl formamide (5ml), and under argon gas, stir and spend the night.Mixture is by water (50ml), 0.5N aqueous hydrochloric acid (50ml), water (50ml), saturated sodium bicarbonate aqueous solution (50ml) and water (50ml) washing successively then.Make dichloromethane layer by the hydrophobic glass material, and vacuum-evaporation obtain crude product.It further obtains pure N-{[2-chloro-3-(trifluoromethyl) phenyl by automatic HPLC (10 * 0.100g injection) purifying of mass spectrometric detection] methyl }-1,3-dimethyl-5-oxo prolineamide (0.613g).
LC/MS[M+H]
+=349, retention time=2.31,2.38 minute (two kinds of diastereomers).
Use in the aforesaid method 1,3-dimethyl-5-oxo proline(Pro) is prepared as follows:
(i) (R, R, R)-2-hydroxyl firpene-3-ketone (10.9g, 64.8mmol) and glycine tertiary butyl ester (13g, 97.2mmol) dry toluene (200ml) solution (0.460g 3.24mmol) handles, then reflux 6 hours under the argon gas with boron trifluoride-ether compound.Then mixture is cooled to room temperature and stirs and spend the night.Filter by sinter (sinter), evaporation obtains yellow jelly then, it is by automatically quick silica gel column chromatography (using Biotage SP4), with the mixture wash-out purifying of 25% ethyl acetate in hexane, obtain some pure N-[(1R, 2R, 5R)-2-hydroxyl-2,6,6-trimethylammonium dicyclo [3.1.1] heptan-3-subunit] glycine 1,1-dimethyl ethyl ester (3.68g) and some blended fractions.Impure material reuses automatically silica gel column chromatography (Biotage SP4) purifying fast, but be to use the hexane solution (0-15% of 0-25% gradient ethyl acetate, surpass 10 times of column volumes, and 15-25%, surpass 5 times of column volumes) wash-out, obtain the pure N-[(1R of another batch, 2R, 5R)-2-hydroxyl-2,6,6-trimethylammonium dicyclo [3.1.1] heptan-3-subunit] glycine 1,1-dimethyl ethyl ester (1.73g).Merge two crowdes of pure N-[(1R, 2R, 5R)-2-hydroxyl-2,6, and 6-trimethylammonium dicyclo [3.1.1] heptan-3-subunit] glycine 1,1-dimethyl ethyl ester (5.41g), this material is used for next step.
(annotate: the salt of wormwood of also available glycine tert-butyl hydrochloride of the tert-butyl glycinate of above-mentioned use and molar equivalent replaces)
(ii) with N-[(1R, 2R, 5R)-2-hydroxyl-2,6,6-trimethylammonium dicyclo [3.1.1] heptan-3-subunit] glycine 1,1-dimethyl ethyl ester (11.05g, anhydrous tetrahydro furan 39.3mmol) (100ml) solution is cooled to-30 ℃, and with ether (17.1ml, 51.1mmol) solution-treated of 3M methyl-magnesium-bromide.Add 1 then, (7.78g, 51.1mmol), and mixture stirred 20 minutes in-30 ℃ 8-diazabicyclo [5.4.0] 11 carbon-7-alkene again.Mixture is handled with ethyl crotonate then, and continues to stir 1 hour.By adding saturated aqueous ammonium chloride (35ml) cancellation mixture, use ethyl acetate (3 * 100ml) extractions then.The organic extract liquid that merges is through dried over sodium sulfate, and filtration and evaporation obtain yellow oil.This material is by automatically quick silica gel column chromatography (using Biotage SP4), with gradient 0-20% (surpass 5 times of column volumes) then the hexane wash-out purifying of 20-35% (surpassing 14 times of column volumes) ethyl acetate obtain N-[(1R, 2R, 5R)-2-hydroxyl-2,6,6-trimethylammonium dicyclo [3.1.1] heptan-3-subunit]-3-methyl L-glutamic acid 1-(1, the 1-dimethyl ethyl) 5-ethyl ester (4.2g), it is used for next step.
(iii) with 10% citric acid (11ml, 5.6mmol) aqueous solution is added to N-[(1R, 2R, 5R)-2-hydroxyl-2,6,6-trimethylammonium dicyclo [3.1.1] heptan-3-subunit]-tetrahydrofuran (THF) (10ml) solution of 3-methyl L-glutamic acid 1-(1, the 1-dimethyl ethyl) 5-ethyl ester (2g) in, and mixture was in stirring at room 4 days.Evaporating mixture, and resistates is suspended in the water (50ml), and wash with ether (100ml).Use sodium bicarbonate aqueous solution to regulate water then, use ether (3 * 100ml) extractions then to pH～7.Merge organic fraction, through dried over sodium sulfate, filtration and evaporation obtain 3-methyl L-glutamic acid 1-(1, the 1-dimethyl ethyl) 5-ethyl ester (1.1g), are yellow oil, and it is not further purified and just is used for next step.
(iv) (1.1g 4.5mmol) places high vacuum to spend the night, and spends weekend then with 3-methyl L-glutamic acid 1-(1, the 1-dimethyl ethyl) 5-ethyl ester.Therefore at this moment initial substance still clearly adds toluene (30ml), and with the gained mixture in 110 ℃ of heated overnight.Evaporation obtains 3-methyl-5-oxo proline(Pro) 1,1-dimethyl ethyl ester (0.79g), and it is not further purified and just is used for next step.
(v) with 3-methyl-5-oxo proline(Pro) 1, (0.79g 3.96mmol) is dissolved in the tetrahydrofuran (THF) (8ml) 1-dimethyl ethyl ester, and (0.27ml 4.36mmol) handles with methyl iodide.Mixture is cooled to 0 ℃ then, and (60% in oil, 0.170g, 4.36mmol) batch treatment with sodium hydride.Stop to produce bubble in 0 ℃ of mixture after 30 minutes, be warmed to room temperature then and stir and spend the night.By adding saturated aqueous ammonium chloride (10ml) cancellation mixture, and separate organic layer and set aside.(3 * 20ml) extract water layer, and the extraction liquid that merges uses hydrophobic glass material drying with methylene dichloride.Merge all organic fractions (comprise before set aside that part), and evaporation obtain thick 1,3-dimethyl-5-oxo proline(Pro) 1,1-dimethyl ethyl ester (0.770g) is yellow jelly, it is not further purified promptly and uses.
(vi) with 1,3-dimethyl-5-oxo proline(Pro) 1, (0.770g 3.62mmol) is suspended in methylene dichloride (5ml) to 1-dimethyl ethyl ester, and (0.4ml 5.4mmol) handles with trifluoroacetic acid.Stirred the mixture 5 hours, then evaporation.With gained resistates and methylbenzene azeotropic, obtain unreacted initial substance (0.600g) then.It is dissolved in the methylene dichloride (2ml) once more, and uses trifluoroacetic acid (2ml) to handle once more.Stir after 2 hours, evaporating mixture, and resistates once more with toluene (10ml) azeotropic obtain thick 1,3-dimethyl-5-oxo proline(Pro) (0.760g), it uses without any being further purified promptly.
Embodiment 130N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-4,4-dimethyl-5-oxo prolineamide (E130)
In room temperature with 1-ethyl-4,4-dimethyl-5-oxo proline(Pro) (0.130g, 0.702mmol, be prepared as follows), I-hydroxybenzotriazole (0.161g, 1.053mmol) and N-(3-dimethylaminopropyl)-N '-ethyl-carbodiimide hydrochloride (0.202g, 1.053mmol) be dissolved in the methylene dichloride (5ml), and stirred 15 minutes.Then with [(2-chloro-4-fluorophenyl) methyl] amine (0.134g, 0.842mmol) and diisopropylethylamine (0.184ml 1.053mmol) is added in the mixture, and continue to stir in room temperature and to spend the night.Vacuum concentrated mixture then, and resistates is allocated between ethyl acetate and the water, and use ethyl acetate extraction.The organic layer that merges is used 3N citric acid, water, saturated aqueous sodium carbonate, water (x3) and salt water washing successively, then through anhydrous sodium sulfate drying.Concentrate and obtain thick solid, it by the automatic HPLC purifying of mass spectrometric detection, obtains N-[(2-chloro-4-fluorophenyl subsequently) methyl]-1-ethyl-4,4-dimethyl-5-oxo prolineamide (0.146g) is solid.LC/MS [M+H]
+=327/329, retention time=2.35 minute.
The 1-ethyl-4 that uses in the aforesaid method, 4-dimethyl-5-oxo proline(Pro) is prepared as follows:
(i) (7.51g 20mmol) is dissolved in the dimethyl formamide (25ml), and with sodium hydride (60% suspension in oil, 0.880g, 22mmol) batch treatment with (S)-(+)-L-5-trityl oxygen ylmethyl-2-Pyrrolidone in 0 ℃.Mixture was in 0 ℃ of stirring 1 hour, and (1.78ml 22mmol) handles to use iodoethane then.Mixture is warmed to room temperature, stirs then and spend the night.Then mixture is poured onto on ice, and extracts with ethyl acetate (x3).The organic extract liquid that merges is water, 50% sodium chloride aqueous solution (3x) and saturated sodium-chloride water solution washing successively, then through dried over sodium sulfate.Concentrate and obtain thick solid, it is by automatically quick silica gel column chromatography (Biotage SP4), and the hexane solution wash-out purifying with 0-100% gradient ethyl acetate obtains pure 1-ethyl-5-{[(trityl group) the oxygen base] methyl }-2-Pyrrolidone (7.09g).
(ii) in-78 ℃ of tetrahydrofuran (THF) (1.912ml to 2M diisopropylaminoethyl lithium, 3.82mmol) drip 1-ethyl-5-{[(trityl group in the solution) the oxygen base] methyl-2-Pyrrolidone (1.34g, 3.48mmol) tetrahydrofuran (THF) (10ml) solution, and the gained mixture stirred 1 hour in-78 ℃.(0.239ml 3.82mmol), and after 1 hour, was warmed to room temperature through 3 hours with mixture in-78 ℃ of restir to add methyl iodide then.Then mixture is cooled to-78 ℃ again, and drips tetrahydrofuran (THF) (1.912ml, 3.82mmol) solution-treated of another part 2M diisopropylaminoethyl lithium.After 1 hour, mixture is used methyl iodide once more, and (0.239ml, 3.82mmol) processing is warmed to mixture room temperature then and stirring is spent the night in-78 ℃ of restir.Ethyl acetate (2x) extraction is used in mixture saturated aqueous ammonium chloride cancellation then.Organic extract liquid water (3x) washing that merges then, then with the saturated sodium-chloride water solution washing, through anhydrous sodium sulfate drying, and the concentrated thick oily solid (1.7g) that obtains.This thick solid is by automatically quick silica gel column chromatography (Biotage SP4), hexane solution wash-out purifying with 0-100% gradient ethyl acetate, obtain the pure products fraction (promptly, 1-ethyl-3,3-dimethyl-5-{[(trityl group) oxygen base] methyl }-2-Pyrrolidone (0.468g)), and the material of pure monoalkylation (that is 1-ethyl-3-methyl-5-{[(trityl group) oxygen base] methyl }-2-Pyrrolidone (0.524g)) and the two mixture (0.240g).Required product is set aside, and merges 1-ethyl-3-methyl-5-{[(trityl group) the oxygen base] methyl }-2-Pyrrolidone and blended material, and be dissolved in the tetrahydrofuran (THF) (20ml).(1.912ml 3.82mmol) in the solution, and continues to stir 1 hour in this temperature in-78 ℃ this drips of solution to be added to the tetrahydrofuran (THF) of 2M diisopropylaminoethyl lithium then.(0.239ml 3.82mmol) is added in the mixture, and makes mixture be warmed to room temperature through 4 hours with methyl iodide then.Obtain another batch 1-ethyl-3 as above-mentioned aftertreatment, 3-dimethyl-5-{[(trityl group) oxygen base] methyl }-2-Pyrrolidone (0.846g), be oily matter, (total mass=1.08g), it is not further purified and just is used for next step with the material merging that had before set aside with it.
(iii) with Amberlyst
(5.56g 26.1mmol) uses methanol wash 3 times, adds 1-ethyl-3 then, 3-dimethyl-5-{[(trityl group) oxygen base] methyl }-2-Pyrrolidone (1.08g, methyl alcohol 2.61mmol) (50ml) solution.Mixture was placed 4 days in room temperature, removed resin (using methanol wash) by filtration then.Concentrate the methyl alcohol fraction that merges, obtain thick oily matter (1.62g), it is by automatically quick silica gel column chromatography (Biotage SP4), hexane solution wash-out purifying with 0-100% gradient ethyl acetate, obtain pure 1-ethyl-5-(hydroxymethyl)-3,3-dimethyl-2-Pyrrolidone (0.376g) is oily matter, places to solidify.
(iv) with 1-ethyl-5-(hydroxymethyl)-3, and 3-dimethyl-2-Pyrrolidone (0.366g, 2.1mmol), Textone (0.387g, 4.3mmol) and 1M SODIUM PHOSPHATE, MONOBASIC aqueous buffer solution (2.46ml, 2.46mmol) be incorporated in the acetonitrile (3ml), and be heated to 40 ℃.TEMPO (2,2,6,6-tetramethyl--piperidino oxygen base free radical) and about 1 SYNTHETIC OPTICAL WHITNER (chlorine bleach liquor, available chlorine＞12%) with some crystalline state is added to mixture then, and continues to stir 4 hours in 40 ℃.Then mixture is poured onto and contains the 1%w/w S-WAT on ice, and the gained mixture extracts with ethyl acetate (x3).The organic extract liquid that merges washs with saturated sodium-chloride water solution, then through dried over mgso, and concentrates and obtains 1-ethyl-4, and 4-dimethyl-5-oxo proline(Pro) (0.392g) is white solid, and it is not further purified promptly and uses.LC/MS[M+H]
+＝186。
Embodiment 131N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-ethyl-4,4-dimethyl-5-oxo prolineamide (E131)
With with above-mentioned synthetic N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-4,4-dimethyl-5-oxo prolineamide (E130) similar methods prepares N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-ethyl-4,4-dimethyl-5-oxo prolineamide replaces [(2-chloro-4-fluorophenyl) methyl] amine except using { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine.
LC/MS[M+H]
+=377/379, retention time=2.63 minute.
Embodiment 132N-[(2-chloro-3, the 4-difluorophenyl) methyl]-1-ethyl-4,4-dimethyl-5-oxo prolineamide (E132)
With with synthetic above-mentioned N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-4,4-dimethyl-similar mode of 5-oxo prolineamide (E130) prepares N-[(2-chloro-3, the 4-difluorophenyl) methyl]-1-ethyl-4,4-dimethyl-5-oxo prolineamide, except using [(2-chloro-3,4-difluorophenyl) methyl] amine hydrochlorate (as above embodiment 36 preparations) to replace [(2-chloro-4-fluorophenyl) methyl] amine.
LC/MS[M+H]
+=345/347, retention time=2.43 minute.
Embodiment 133N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-5-oxo-4,4-two (phenyl methyl) prolineamide (E133)
With with above-mentioned synthetic N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-4,4-dimethyl-5-oxo prolineamide (E130) similar methods prepares N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-5-oxo-4, two (phenyl methyl) prolineamides of 4-, except using 1-ethyl-5-oxo-4,4-two (phenyl methyl) proline(Pro) replaces 1-ethyl-4,4-dimethyl-5-oxo proline(Pro).With with above-mentioned synthetic embodiment 130 in 1-ethyl-4,4-dimethyl-5-oxo proline(Pro) similar methods prepares 1-ethyl-5-oxo-4,4-two (phenyl methyl) proline(Pro), except using 1-ethyl-3,3-two (phenyl methyl)-5-{[(trityl group) oxygen base] methyl }-2-Pyrrolidone is (as the separation of by-products among the method B, embodiment 37) replacement 1-ethyl-3,3-dimethyl-5-{[(trityl group) the oxygen base] methyl }-2-Pyrrolidone.
LC/MS[M+H]
+=479/481, retention time=3.32 minute.
Embodiment 134N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-ethyl-5-oxo-4-(phenyl methyl) prolineamide (E134)
With with above-mentioned synthetic N-[(2-chloro-4-fluorophenyl) methyl]-1-ethyl-5-oxo-4-(phenyl methyl)-prolineamide (E37) similar methods prepares N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl-1-ethyl-5-oxo-4-(phenyl methyl) prolineamide, replace [(2-chloro-4-fluorophenyl) methyl] amine except using { [2-chloro-3-(trifluoromethyl) phenyl] methyl } amine.Using method B (as described in example 37 above) preparation 1-ethyl-5-oxo-4-(phenyl methyl)-proline(Pro).
LC/MS[M+H]
+=439/441, retention time=2.99 minute.
Embodiment 135N-{[2-cyano group-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo prolineamide (E135)
With with above-mentioned synthetic N-{[2-cyano group-3-(trifluoromethyl) phenyl] methyl-1-ethyl-5-oxo prolineamide (E105) similar methods prepares N-{[2-cyano group-3-(trifluoromethyl) phenyl] methyl-1-methyl-5-oxo prolineamide, replace 1-ethyl-5-oxo-proline(Pro) except using 1-methyl-5-oxo-proline(Pro).
LC/MS[M+H]
+=326, retention time=2.02 minute.
Embodiment 136N-(2-xenyl methyl)-1-ethyl-5-oxo prolineamide (E136)
With with above-mentioned synthetic N-[(2, the 3-3,5-dimethylphenyl) methyl]-1-ethyl-5-oxo prolineamide (E50) similar methods prepares N-(2-xenyl methyl)-1-ethyl-5-oxo prolineamide, except using (2-xenyl methyl) amine to replace 2, the 3-dimethyl benzyl amine.
LC/MS[M+H]
+=323, retention time=2.38 minute.
Microwave reactor
When having in the foregoing description when indicated, used microwave reactor is Biotage Initiator
TMUnless otherwise indicated, use normal power output to react.
The automatic HPLC of mass spectrometric detection
When having in the foregoing description when indicated, the purifying of the automatic HPLC by mass spectrometric detection uses following equipment and condition to carry out:
Hardware
Waters?2525Binary?Gradient?Module
Waters?515Makeup?Pump
Waters?Pump?Control?Module
Waters?2767Inject?Collect
Waters?Column?Fluidics?Manager
Waters?2996Photodiode?Array?Detector
Waters?ZQ?Mass?Spectrometer
Gilson?202fraction?collector
Gilson?Aspec?waste?collector
Software
Waters?MassLynx?version?4SP2
Pillar
Used pillar is Waters Atlantis, and it is of a size of 19mm * 100mm (on a small scale) and 30mm * 100mm (on a large scale).The stationary phase particle diameter is 5 μ m.
Solvent
A: aqueous solvent=water+0.1% formic acid
B: organic solvent=acetonitrile+0.1% formic acid
Aftertreatment (make up) solvent=methyl alcohol: water 80: 20
Pin cleaning solvent=methyl alcohol
Method
According to the analysis retention time of concern compound, use 5 kinds of methods.They have 13.5-minute working time, comprise 10-minute gradient, then the step of post flushing in 3.5 minutes and reequilibrate.
Greatly/small-scale 1.0-1.5=5-30%B
Greatly/small-scale 1.5-2.2=15-55%B
Greatly/small-scale 2.2-2.9=30-85%B
Greatly/small-scale 2.9-3.6=50-99%B
Greatly/small-scale 3.6-5.0=80-99%B (6 minutes, washed and reequilibrate in 7.5 minutes then)
Flow velocity
The flow velocity of all aforesaid methods is 20mls/ minute (on a small scale) or 40mls/ minute (on a large scale).
Chirality HPLC
The equipment and the condition of enantiomeric purity that is used to characterize selected sample is as follows:
Method (A)
Instrument: Agilent 1100Series Liquid Chromatogram
Pillar: Chiralpak AD (250mm * 4.6mm; The 10um particle diameter)
Moving phase: heptane: dehydrated alcohol (70: 30) v/v pump mixes
Flow velocity: 1ml/ minute
Temperature: envrionment temperature
U.V. wavelength: 215nm
Method (B)
Instrument: Agilent 1100Series Liquid Chromatogram
Pillar: Chiralpak AD (250mm * 4.6mm; The 10um particle diameter)
Moving phase: heptane: dehydrated alcohol (50: 50) v/v pump mixes
Flow velocity: 1ml/ minute
Temperature: envrionment temperature
U.V. wavelength: 215nm
Method (C)
Instrument: Agilent 1100Series Liquid Chromatogram
Pillar: Chiralpak AD (250mm * 4.6mm; The 10um particle diameter)
Moving phase: heptane: dehydrated alcohol (80: 20) v/v pump mixes
Flow velocity: 1ml/ minute
Temperature: envrionment temperature
U.V. wavelength: 215nm
Method (D)
Instrument: Agilent 1100Series Liquid Chromatogram
Pillar: Chiralpak AS (250mm * 4.6mm; The 10um particle diameter)
Moving phase: heptane: dehydrated alcohol (80: 20) v/v pump mixes
Flow velocity: 1ml/min
Temperature: envrionment temperature
U.V. wavelength: 215nm
Liquid chromatography/mass spectrometry
The above-mentioned embodiment that analyzes by liquid chromatography/mass spectrometry (LC/MS) uses following equipment and condition to carry out:
Hardware
Agilent 1100 Gradient pumps
Agilent 1100 automatic samplers
Agilent 1100DAD detector
Agilent 1100 degassers
Agilent 1100 baking ovens
Agilent 1100 controllers
Waters ZQ mass spectrograph
Sedere?Sedex?85
Software
Waters?MassLynx?version?4.0SP2
Pillar
Used pillar is Waters Atlantis, and it is of a size of 4.6mm * 50mm.The stationary phase particle diameter is 3 μ m.
Solvent
A: water-containing solvent=water+0.05% formic acid
B: organic solvent=acetonitrile+0.05% formic acid
Method
Used general method has 5 minute working time.
| Time/minute | ％B |
| 0 | 3 |
| 0.1 | 3 |
| 4 | 97 |
| 4.8 | 97 |
| 4.9 | 3 |
| 5.0 | 3 |
The flow velocity of aforesaid method is 3ml/ minute.
The sampling volume of general method is 5ul.
Column temperature is 30 degree.
The UV sensing range is 220 to 330nm.
Pharmacology data
Can detect the external biological activity of The compounds of this invention according to following research to the P2X7 acceptor:
Ethidium accumulation experiment
Use the NaCl experiment damping fluid of following composition (mM of unit) to study: 140mM NaCl, HEPES 10, N-methyl D-glucosamine 5, KCl 5.6, D-glucose 10, CaCl
2(0.5 pH 7.4).The HEK293 cell of expressing human reorganization P2X7 acceptor was grown 18-24 hour in 96 orifice plates of handling with poly-L-Lysine in advance.(clone of people P2X7 acceptor is described in US 6,133,434 in).This cell adds 50 μ l testing compounds then with 350 μ l experiment damping fluid washing 2 times.Cell was hatched 30 minutes in room temperature (19-21 ℃) then, added ATP and ethidium (the final experimental concentrations of 100 μ M) then.Select the EC of ATP concentration near this acceptor type
80, and be 1mM for the people P2X7 acceptor of being studied.Continued to hatch 8 or 16 minutes, and stop by the 1.3M sucrose (the P2X7 receptor antagonist that contains 5mM, reactive black (reactive black) 5 (Aldrich)) that adds 25 μ l.By using CanberraPackard Fluorocount (Pangbourne, UK) or Flexstation.II (Molecular Devices) detect cell accumulation (excitation wavelength: 530nm, and emission wavelength: 620nm) from the fluorometric assay ethidium at the bottom of the plate.Use the iteration curve fitting technique, detect antagonist pIC50 value for blocking-up ATP response.
The fluorescence imaging plate reads (FLIPR) Ca experiment
Use the NaCl experiment damping fluid of following composition (mM of unit) that people P2X7 is studied: 137NaCl; 20HEPES; 5.37KCl; 4.17NaHCO
31CaCl
20.5MgSO
4D-glucose (pH 7.4) with 1g/L.
Expressing human is recombinated the HEK293 cell of P2X7 acceptor with growth in poly-L-Lysine pretreated 384 orifice plates 42-48 hour.(clone of people P2X7 acceptor is described in US 6,133,434 in).This cell adds 2 μ M Fluo4 (Teflabs) in 37 ℃ and kept 1 hour with 80 μ l experiment damping fluid washing 3 times, washs 3 times once more, and keeps 30 μ l damping fluids, adds the testing compound of the 4x concentration of 10 μ l then.Then with cell in incubated at room 30 minutes, add Benzoylbenzoyl-ATP (BzATP) (online) the final experimental concentration of 60 μ M then by FLIPR384 or FLIPR3 instrument (molecular device).Select BzATP concentration with approaching EC for this receptor type
80Continue to hatch and reading 90 seconds, detect from the fluorescence at the bottom of the plate (excitation wavelength: 488nm, and emission wavelength: 516nm) measure the increase of intracellular Ca2+ by using FLIPR CCD photographic camera.Use the iteration curve fitting technique, detect antagonist pIC for blocking-up BzATP response
50Value.
In FLIPR Ca experiment and/or ethidium accumulation experiment, detect the antagonistic activity of the compound of embodiment 1-136, and find pIC50 value＞4.7 in FLIPR Ca experiment people P2X7 acceptor, and/or pIC50 value＞5.5 in ethidium accumulation experiment.
Experiment in the body
The rat model of neuropathic pain
By in the total neural loose narrow ligation on every side of ischium, produce mononeuropathy on every side, produce the rat model of neuropathic pain thus, people such as Bennet, Pain, Vol.33, pp87-107 (1988).From Charles River, the bull Random Hooded rat (180-200g) of UK is anaesthetized with isoflurane (3%).Big midleg at left leg exposes sciatic nerve, and as people such as Bennet, Pain, Vol.33, described in the pp87-107 (1988) with 4 loose ligature Chromic 4.0 gutstring ligation nerves.Sew up a wound and tight with linear system.The identical process of rat experience of sham operated rats is not except using loose ligature.Use the generation of handbook applicating evaluating machinery (sense of touch) allodynia of Von Frey hair monofilament.Up direction uses the plantar region (scope: 1.4g-26g) of monofilament to back leg.Every hair uses about 3-5 second up to observing the claw withdrawing reaction.After reusing lower and/or higher hair affirmation, record produces the minimum hair of claw withdrawal as threshold response (g).When setting up stable allodynia, gave totally 8 days every day 2 times of drug compound to rat in postoperative 26-33 days, recording exceptional pain detects 3 times at least during the administration.Compare N-[(2,4-dichlorophenyl with solvent response) methyl]-1-methyl-5-oxo-prolineamide (E10) and N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo prolineamide (E51) mechanical allodynia of CCI-inductive that obviously reversed.
The rat model of arthralgia
By detecting, can in the arthralgia model of chronic inflammatory pain, estimate potential analgesic effect in the allergy of inversed F CA-inductive with the allergy of FCA intra-articular injection behind the knee.Will be from Charles River, the bull Random Hooded rat (150-180g) of UK is used isoflurane (3%) anesthesia simply.Inject the freund complete adjuvant (FCA) of 150 μ l to left knee joint (intraarticular i.art) to rat then.Before using Dual Channel Weight Averager (Linton Instruments) operation and measure afterwards the weighing load of each hind leg ability (weight loading, g).When injection and the offside pawl between when setting up the stable difference of weight loading, generally, totally 5 days, write down the weight loading measuring result every day to rat oral administration compound (being generally operation back 13-17 days) every day 2 times.Compare N-[(2 with the solvent response, the 4-dichlorophenyl) methyl]-1-methyl-5-oxo-prolineamide (E10) and N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-the obvious inversed F CA (i.art) of 1-methyl-5-oxo prolineamide (E51)-inductive weight loading difference, and produce the ED50 ' s＜20mg/kg that calculates from area under curve (AUC).
The rat model of acute inflammation pain
The useful animal model that is used for acute inflammation pain is freund complete adjuvant (FCA)-inductive inflammatory model.Use the similar model of carrageenan rather than FCA to be described in people .Br.J.Pharmacol.1997 such as Clayton; 120, among the 219P.From Charles River, the bull RandomHooded rat (180-220g) of UK is accepted the plantar surface of the FCA of foot pad (i.pl) injection 100 μ l to the left side rear solid end.Using the ability of Dual Channel Weight Averager (Linton Instruments) measurement before the FCA injection and after injecting 24 hours to the weighing load of each hind leg.Behind the reading, generally, write down the weight loading measuring result afterwards behind the FCA to rat oral administration compound.Compare with the solvent response, N-[(2, the 4-dichlorophenyl) methyl]-1-methyl-5-oxo-prolineamide (E10) and N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-the obvious inversed F CA (i.pl) of 1-methyl-5-oxo prolineamide (E51)-inductive weight loading difference, and produce the ED50 ' s＜20mg/kg that calculates from dose response curve.
Claims (25)
1. formula (I) compound or its pharmacy acceptable salt:
Wherein:
R
1Represent unsubstituted methyl, ethyl, C
3-5Cycloalkyl, pyridylmethyl-, phenyl or benzyl;
R
2And R
3Represent hydrogen, halogen, C independently
1-6Alkyl or benzyl; And any described C
1-6Alkyl or benzyl are optional to be replaced by 1,2 or 3 halogen atom;
R
4, R
5And R
6Represent hydrogen, fluorine or methyl independently; And
R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, C independently
1-6Alkyl or phenyl, and any described C
1-6Alkyl or phenyl is optional to be replaced by 1,2 or 3 halogen atom; Or R
10And R
11Form the optional phenyl ring that is replaced by 1,2 or 3 halogen atom with the carbon atom that links to each other with them;
Condition is to work as R
7And R
11When all being selected from hydrogen or fluorine, R
8, R
9And R
10In at least one is a halogen atom, or R
8, R
9And R
10Be selected from hydrogen and CF
3, and R
8, R
9And R
10In one but at the most one be CF
3
2. formula (I) compound or its pharmacy acceptable salt:
Wherein:
R
1Represent unsubstituted methyl, ethyl, C
3-5Cycloalkyl, phenyl or benzyl;
R
2And R
3Represent hydrogen, halogen, C independently
1-6Alkyl or benzyl; And any described C
1-6Alkyl or benzyl can be chosen wantonly by 1,2 or 3 halogen atom and replace;
R
4, R
5And R
6Represent hydrogen or fluorine independently; And
R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, C independently
1-6Alkyl or phenyl; And any described C
1-6Alkyl or phenyl can be chosen wantonly by 1,2 or 3 halogen atom and replace;
Condition is to work as R
7And R
11When representing hydrogen or fluorine independently, R
8, R
9And R
10In at least one is a halogen atom.
3. formula (I) compound or its salt of claim 1 definition, wherein R
1Expression methyl or ethyl.
4. formula (I) compound or its salt of claim 1 definition, wherein R
2And R
3Represent hydrogen, fluorine or methyl independently.
5. formula (I) compound or its salt of claim 3 definition, wherein R
2And R
3Represent hydrogen, fluorine or methyl independently.
6. formula (I) compound or its salt of claim 1,3 or 4 each definition, wherein R
4, R
5And R
6Represent hydrogen or methyl independently.
7. formula (I) compound or its salt of claim 1,3,4 or 5 each definition, wherein R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, trifluoromethyl or unsubstituted C independently
1-6Alkyl; Or R
10And R
11Form unsubstituted phenyl ring with the carbon atom that links to each other with them.
8. formula (I) compound or its salt of claim 6 definition, wherein R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, trifluoromethyl or unsubstituted C independently
1-6Alkyl; Or R
10And R
11Form unsubstituted phenyl ring with the carbon atom that links to each other with them.
9. formula (I) compound or its salt of claim 1 definition, wherein R
7, R
8, R
9, R
10And R
11Represent hydrogen, chlorine, fluorine, bromine, methyl or trifluoromethyl independently.
10. formula (I) compound or its pharmacy acceptable salt of claim 1 definition:
Wherein, R
1Represent unsubstituted methyl, ethyl, C
3-5Cycloalkyl, pyridylmethyl-, phenyl or benzyl;
R
2And R
3All represent hydrogen;
R
4, R
5And R
6Represent hydrogen or methyl independently; And
R
7, R
8, R
9, R
10And R
11Represent hydrogen, chlorine, fluorine, bromine, methyl or trifluoromethyl independently;
Condition is to work as R
7And R
11When all being selected from hydrogen or fluorine, R
8, R
9And R
10In at least one is a halogen atom, or R
8, R
9And R
10Be selected from hydrogen and CF
3, and R
8, R
9And R
10In one but at the most one be CF
3
11. the compound or its salt of claim 10 definition, wherein R
1Expression methyl or ethyl.
12. formula (I) compound or its salt of claim 1 definition, it is compound or its pharmacy acceptable salt as described below:
13. formula (I) compound or its salt of claim 1 definition, it is N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo prolineamide or its pharmacy acceptable salt.
14. formula (I) compound or its salt of claim 13 definition, it is N-{[2-chloro-3-(trifluoromethyl) phenyl] methyl }-1-methyl-5-oxo-L-prolineamide or its pharmacy acceptable salt.
15. formula (I) compound or its salt of claim 1 definition, it is N-[(2, the 4-dichlorophenyl) methyl]-1-methyl-5-oxo-prolineamide or its pharmacy acceptable salt.
16. the compound or its salt of claim 15 definition, it is N-[(2, the 4-dichlorophenyl) methyl]-1-methyl-5-oxo-L-prolineamide or its pharmacy acceptable salt.
17. formula (I) compound or its pharmacy acceptable salt are used for the treatment of purposes in the medicine of the receptor-mediated pain of P2X7, inflammation or neurodegenerative disease in preparation:
Wherein:
R
1Represent unsubstituted C
1-6Alkyl, C
2-6Alkenyl, C
3-6Cycloalkyl or pyridylmethyl, phenyl or benzyl;
R
2And R
3Represent hydrogen, halogen, C independently
1-6Alkyl or benzyl; And any described C
1-6Alkyl or benzyl are optional to be replaced by 1,2 or 3 halogen atom;
R
4, R
5And R
6Represent hydrogen, fluorine or methyl independently; And
R
7, R
8, R
9, R
10And R
11Represent hydrogen, halogen, cyano group, C independently
1-6Alkyl or phenyl, and any described C
1-6Alkyl or phenyl is optional to be replaced by 1,2 or 3 halogen atom; Or R
10And R
11Form the optional phenyl ring that is replaced by 1,2 or 3 halogen atom with the carbon atom that links to each other with them;
Condition is to work as R
7And R
11When all being selected from hydrogen or fluorine, R
8, R
9And R
10In at least one is a halogen atom, or R
8, R
9And R
10Be selected from hydrogen and CF
3, and R
8, R
9And R
10In one but at the most one be CF
3
18. the purposes of formula (I) compound or its salt of claim 17 definition, wherein R
1Represent unsubstituted C
1-4Alkyl, C
3-5Cycloalkyl, pyridylmethyl-, phenyl or benzyl.
19. the purposes of formula (I) compound or its salt of claim 17 definition:
Wherein, R
1Represent unsubstituted C
1-4Alkyl, C
2-4Alkenyl, C
3-5Cycloalkyl, pyridylmethyl-, phenyl or benzyl;
R
2And R
3All represent hydrogen;
R
4, R
5And R
6Represent hydrogen or methyl independently; And
R
7, R
8, R
9, R
10And R
11Represent hydrogen, chlorine, fluorine, bromine, methyl or trifluoromethyl independently;
Condition is to work as R
7And R
11When all being selected from hydrogen or fluorine, R
8, R
9And R
10In at least one is a halogen atom, or R
8, R
9And R
10Be selected from hydrogen and CF
3, and R
8, R
9And R
10In one but at the most one be CF
3
20. the purposes of the formula of each definition of claim 17-19 (I) compound or its salt, wherein this medicine is to be used for the treatment of or to prevent the receptor-mediated inflammatory pain of P2X7, neuropathic pain, chronic arthralgia or visceral pain.
21. the purposes of the formula of each definition of claim 17-19 (I) compound or its salt, wherein this medicine is to be used for the treatment of or to prevent the receptor-mediated degenerative brain disorder of P2X7.
22. pharmaceutical composition, it comprises formula (I) compound or its pharmacy acceptable salt and the pharmaceutically acceptable carrier or the vehicle of each definition of claim 1-16.
23. the formula of each definition (I) compound or its pharmacy acceptable salt are used for the treatment of purposes in the medicine of the receptor-mediated pain of P2X7, inflammation or neurodegenerative disease in preparation among the claim 1-16.
24. (I) compound of each definition or its pharmacy acceptable salt are used for the treatment of or prevent purposes in the medicine of the receptor-mediated inflammatory pain of P2X7, neuropathic pain, chronic arthralgia or visceral pain in preparation among the claim 1-16.
25. (I) compound of each definition or its pharmacy acceptable salt are used for the treatment of or prevent purposes in the medicine of the receptor-mediated degenerative brain disorder of P2X7 in preparation among the claim 1-16.
Applications Claiming Priority (9)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB0613473.8 | 2006-07-06 | ||
| GB0613473A GB0613473D0 (en) | 2006-07-06 | 2006-07-06 | Novel compounds |
| GB0622825A GB0622825D0 (en) | 2006-11-15 | 2006-11-15 | Novel compounds |
| GB0622825.8 | 2006-11-15 | ||
| GB0705263.2 | 2007-03-19 | ||
| GB0705263A GB0705263D0 (en) | 2007-03-19 | 2007-03-19 | Novel compounds |
| GB0711439.0 | 2007-06-13 | ||
| GB0711439A GB0711439D0 (en) | 2007-06-13 | 2007-06-13 | Novel receptor antagonists and their methods of use |
| PCT/EP2007/056675 WO2008003697A1 (en) | 2006-07-06 | 2007-07-03 | Substituted n-phenylmethyl -5-oxo-proline-2-amides as p2x7-receptor antagonists and their methods of use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101511787A CN101511787A (en) | 2009-08-19 |
| CN101511787B true CN101511787B (en) | 2011-11-30 |
Family
ID=36926597
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007800330531A Expired - Fee Related CN101511787B (en) | 2006-07-06 | 2007-07-03 | Substituted N-phenylmethyl -5-oxo-proline-2-amides as P2X7-receptor antagonists and their methods of use |
Country Status (4)
| Country | Link |
|---|---|
| CN (1) | CN101511787B (en) |
| AP (1) | AP2009004734A0 (en) |
| GB (1) | GB0613473D0 (en) |
| ZA (1) | ZA200810726B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109180552B (en) * | 2018-09-26 | 2021-10-26 | 沈阳药科大学 | Substituted 5-oxo pyrrolidine derivative and preparation method and application thereof |
| WO2022161423A1 (en) * | 2021-01-28 | 2022-08-04 | 深圳晶泰科技有限公司 | Cyclic amine derivative, preparation method therefor and use thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1284074A (en) * | 1997-12-05 | 2001-02-14 | 阿斯特拉曾尼卡英国有限公司 | Novel compounds |
-
2006
- 2006-07-06 GB GB0613473A patent/GB0613473D0/en not_active Ceased
-
2007
- 2007-07-03 CN CN2007800330531A patent/CN101511787B/en not_active Expired - Fee Related
- 2007-07-03 AP AP2009004734A patent/AP2009004734A0/en unknown
-
2008
- 2008-12-19 ZA ZA200810726A patent/ZA200810726B/en unknown
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1284074A (en) * | 1997-12-05 | 2001-02-14 | 阿斯特拉曾尼卡英国有限公司 | Novel compounds |
Also Published As
| Publication number | Publication date |
|---|---|
| GB0613473D0 (en) | 2006-08-16 |
| CN101511787A (en) | 2009-08-19 |
| AP2009004734A0 (en) | 2009-02-28 |
| ZA200810726B (en) | 2009-12-30 |
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