CN103058949A - Thiazole derivative acting as DHODH inhibitor and its application - Google Patents

Thiazole derivative acting as DHODH inhibitor and its application Download PDF

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CN103058949A
CN103058949A CN2011103171249A CN201110317124A CN103058949A CN 103058949 A CN103058949 A CN 103058949A CN 2011103171249 A CN2011103171249 A CN 2011103171249A CN 201110317124 A CN201110317124 A CN 201110317124A CN 103058949 A CN103058949 A CN 103058949A
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赵振江
韩乐
许鸣豪
高瑞
黄瑾
刁妍妍
曹贤文
崔坤强
李洪林
徐玉芳
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East China University of Science and Technology
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Priority to PCT/CN2012/086816 priority patent/WO2013056684A2/en
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Abstract

The invention relates to a thiazole derivative which is shown as general formula I and acts as a DHODH (dihydroorotate dehydrogenase) inhibitor, and its application. The compound provided in the invention can be used for treatment or prevention of various DHODH-mediated diseases, including but not limited to rheumatoid arthritis, tumors, organ transplantation rejection, psoriasis and other autoimmune diseases.

Description

Thiazole derivative and application thereof as the DHODH inhibitor
Technical field
The invention belongs to pharmaceutical chemistry and pharmacotherapeutics field, be specifically related to thiazole derivative and application thereof as the DHODH inhibitor.
Background technology
Dihydroorate dehydrogenase (Dihydroorotate dehydrogenase, DHODH) be that catalysis dihydroorotate dehydrogenation makes it be converted into a kind of enzyme of vitamin B13, this process belongs to the four-step reaction of pyrimidine de novo synthesis, thereby DHODH is the synthetic key enzyme of nucleic acid pyrimidine.Suppress DHODH, can block the synthetic of de novo pyrimidine, cause DNA and RNA dyssynthesis.For most of biological, pyrimidine bases can obtain by de novo synthesis and salvage pathway, but the human cell who breaks up fast, the T-lymphocyte, B-lymphocyte and the tumour cell that for example activate, the de novo synthesis that need to depend on pyrimidine satisfies its growth demand.This is so that the DHODH inhibitor can be used as the treatment that cellular antiproliferative agent is used for tumour and some immunosuppression reaction.Because the general mechanism of action that DHODH is synthetic to DNA and RNA, also so that its inhibition many other the signal paths in downstream are exerted an influence.Research (Proceedings of the National Academy of Sciences 2010,107 (29), 12828) shows that the inhibition of DHODH can cause the p53 stress reaction in the plastosome, can be used for the treated tissue damage.Other has document (Annals of the Rheumatic Diseases 2006,65 (6), 728-735) report, the transcription factor NF-KB phosphorylation that the inhibition of DHODH can stop TNF to induce, suppress AP-1 and the kinase whose activation of c-iun N-terminal protein, finally suppress the apoptosis reaction that TNF induces.
Human dihydroorate dehydrogenase is positioned on the mitochondrial inner membrane, and its catalytic process needs the participation of other cofactor.On structure, DHODH mainly is divided into two portions, i.e. the α of the α-helixstructure of N-end zone and C-end/β barrel-like structure.DHODH is attached on the mitochondrial inner membrane by the unique texture of N-end, and the C-stub area then is the main site of katalysis.The binding site that on DHODH, has simultaneously substrate and cofactor vitamin B2 phosphate (Flavin mononucleotide, FMN) and ubiquinone (Coenzyme, CoQ).Its catalytic process is mainly finished by two-step reaction: at first dihydroorotate is reduced to vitamin B13, and simultaneously FMN accepts the hydrogen atom that this process takes off and is oxidized to FMNH 2Afterwards under the effect of CoQ, FMNH 2Dehydrogenation occurs again be reduced to FMN.
Theoretically, any effect that can block with the compound of substrate or cofactor competitive binding DHODH, thus blocking dna or RNA's is synthetic.(Biochemical pharmacology 1988,37 (20), and 3807-3816), such inhibitor competition is combined in the redox site of C-end as the research of DHODH inhibitor report to be arranged once in early days with the dihydroorotate analogue.Now the research of DHODH inhibitor is mainly carried out for the CoQ binding site, wherein Bu Kuina (Brequinar) and leflunomide (Leflunomide) have been applied to clinical.Brequinar is used for host immune response antitumor and that organ transplantation causes, but Brequinar treatment window is narrow, and when combination with cisplatin or cyclosporin A oral administration, cause easily the side effect of thrombopenia, mucositis, limited its widespread use in clinical.Leflunomide was in listing in 1998, acute and chronic reaction and Xeno-rejection reaction that various autoimmune disease, organ transplantation are caused all have very strong restraining effect, be used for the treatment of clinically lupus erythematosus, rheumatoid arthritis, and can be used for preventing and treating the graft rejection reaction.(Nature 2011 for current research, 471 (7339), show that 518-522) Leflunomide is as potent DHODH inhibitor, use separately or all can produce restraining effect to the melanocyte oncocyte of external and experimental mouse with the other medicines coupling.Because side effects such as that the life-time service of Leflunomide also can produce is unusual such as the liver enzyme, hypertension or fash, therefore seek the little efficient DHODH inhibitor of toxic side effect and remain study hotspot in immune correlated disease and the oncotherapy.
Other can use disease that the DHODH inhibitor treats and pertinent literature also can referring to, comprise rheumatoid arthritis (Herrmann, M.L., Schleyerbach, R. and Kirschbaum, B.J., Leflunomide:an immunomodulatory drug for the treatment of rheumatoid arthritis and other autoimmune diseases.Immunopharmacology 2000,47 (2-3), 273-289), colitis (Fitzpatrick, L.R., Deml, L., Hofmann, C.; Small, J.S., Groeppel, M., Hamm, S., Lemstra, S., Leban, J. and Ammendola, A., 4SC-101, a novel immunosuppressive drug, inhibits IL-17 and attenuates colitis in two murine models of inflammatory bowel disease.Inflammatory bowel diseases 2010,16 (10), 1763-1777), systemic lupus erythematous (Kulkarni, O.P., Sayyed, S.G., Kantner, C., Ryu, M., Schnurr, M., Sardy, M., Leban, J., Jankowsky, R., Ammendola, A. and Doblhofer, R., 4SC-101, A Novel Small Molecule Dihydroorotate Dehydrogenase Inhibitor, Suppresses Systemic Lupus Erythematosus in MRL-(Fas) lpr Mice.Am.J.Pathol.2010,176 (6), 2840-2847), psoriatic arthritis (Kaltwasser, J.P., Nash, P., Gladman, D., Rosen, C.F., Behrens, F., Jones, P., Wollenhaupt, J., Falk, F.G. and Mease, P., Efficacy and safety of leflunomide in the treatment of psoriatic arthritis and psoriasis:A multinational, double-blind, randomized, placebo-controlled clinical trial.Arthritis ﹠amp; Rheumatism 2004,50 (6), 1939-1950), psoriatic (White, R.M., Cech, J., Ratanasirintrawoot, S., Lin, C.Y., Rahl, P.B., Burke, C.J., Langdon, E., Tomlinson, M.L., Mosher, J. and Kaufman, C., DHODH modulates transcriptional elongation in the neural crest and melanoma, Nature 2011,471 (7339), 518-522), transplant rejection (Makowka, L.; Sher, L.; Cramer, D.The development of Brequinar as an immunosuppressive drug for transplantation.Immunological reviews 1993,136,51), renal glomerular disease (Ceng Jianying, Zhang Jianlin, leflunomide is used for the clinical observation of renal glomerular disease, Chinese journals of practical medicine, 2006 (15)); Deng.
Summary of the invention
According to the principle that the DHODH inhibitor is combined with the ubiquinone binding pocket that is positioned at DHODH enzyme N-terminal, inhibitor all can have head and hydrophobic afterbody of a polarity, and this structure can be combined in the ubiquinone binding pocket them effectively.Accordingly, medicinal design, pharmaceutical chemistry and molecular biology method and technology are calculated in inventor's integrated use in previous work, a series of thiazole derivatives of said structure requirement have been found to meet, structural framework is different from the high reactivity DHODH inhibitor (leflunomide, Teriflunomide (teriflunomide, A771726), Bu Kuina) that bibliographical information is crossed fully.Some compounds wherein have significant immunosuppressive activity at cell and animal level, and toxicity is very low simultaneously, and security is good, has good patent medicine prospect.For this series lead compound, the inventor designs and has synthesized a series of thiazole derivatives, and general structure is as follows:
Figure BDA0000099751710000031
In the formula,
R 1Be selected from H, C 1-C 6Alkyl, C 2-C 6Alkenyl or alkynyl, the optional aryl that replaces, nitro, amino, NR 4R 5, halogen;
R 2Be selected from H, C 1-C 6Alkyl, the C that halogen replaces 1-C 6Alkyl, C 1-C 6Unsaturated alkyl, C 1-C 3Alkyl-carbonyl, the optional benzoyl that replaces, carboxyl, aminocarboxyl, C 1-C 6Alkoxy carbonyl, hydroxyl, C 1-C 6Alkoxyl group, the optional aryl that replaces, the optional heteroaryl that replaces, amino, NHR 6
R 3Be selected from H, C 1-C 6Alkyl ,-C (O) NHR 7, C 1-C 6Alkoxy carbonyl, the alkyl that halogen replaces, C 2-C 6Alkenyl or alkynyl, the optional phenyl that replaces, C 1-C 4Alkyl-carbonyl, the optional benzoyl that replaces, the optional pyridyl carbonyl that replaces, C 1-C 3Alkyl carboxyl, amide group, amino, C 1-C 10The amino that alkyl replaces, halogen;
R 4And R 5Independently be selected from separately H, the optional aryl that replaces, the optional heterocyclic radical that replaces, the optional aryl carbonyl that replaces, the optional heterocyclic radical carbonyl that replaces, and the optional aryloxy alkyl carbonyl that replaces;
R 6Be selected from C 1-C 6Alkyl, the optional phenyl that replaces, or form 6 Yuans rings with the N that is connected, such as piperidine ring, or 6 Yuans contain oxygen or nitrogen heterocyclic, such as piperazine ring and morpholine ring; With
R 7Be selected from the optional aryl that replaces and the optional heterocyclic radical that replaces.
In a preferred embodiment, described compound is selected from the compound of Formula Il:
Ar is selected from the optional aryl that replaces, the optional heterocyclic radical that replaces, the optional aryl carbonyl that replaces, the optional heterocyclic radical carbonyl that replaces, and the optional aryloxy alkyl carbonyl that replaces;
R 2Be selected from C 1-C 6Alkyl, the C that halogen replaces 1-C 6Alkyl, C 1-C 3Alkyl-carbonyl, the optional benzoyl that replaces, carboxyl, aminocarboxyl, C 1-C 6Alkoxy carbonyl, hydroxyl, C 1-C 6Alkoxyl group, optional aryl or the heteroaryl that replaces, amino, NHR 6
R 3Be selected from C 1-C 6Alkyl, C 1-C 6Alkoxy carbonyl, the optional phenyl that replaces, C 1-C 4Alkyl-carbonyl, the optional benzoyl that replaces, C 1-C 3Alkyl carboxyl, amide group, the optional aniline formyl radical that replaces.
In a preferred embodiment, the substituting group on the Ar comprises C 1-C 10Alkyl, C 3-C 8Cycloalkyl, C 1-C 4Alkoxyl group, the optional phenyl that replaces, the optional phenoxy group that replaces, benzyloxy, CF 3, and halogen, substituent quantity is 1,2,3,4 or 5.Ar can be benzo thick (mixing) cyclic cpds, and heteroatoms comprises N, O, S, Se, such as anthryl, carbazyl, indanyl, tetrahydro naphthyl etc.
In more preferred embodiment, R 2Be selected from C 1-C 6Alkyl, CF 3, phenyl, ethanoyl, benzoyl, carboxyl, formamyl, C 1-C 6Alkoxy carbonyl, amino.Methyl more preferably, CF 3, phenyl.
In more preferred embodiment, R 3Be selected from C 1-C 6Alkyl, phenyl, C 1-C 3Alkyl-carbonyl, the optional benzoyl that replaces, carboxyl, C 1-C 6Alkoxy carbonyl, amide group, the optional aniline formyl radical that replaces; Preferred C 1-C 3Alkyl-carbonyl, C 1-C 6Alkoxy carbonyl.
In a preferred embodiment, described compound is selected from the compound of Formula Il I:
Figure BDA0000099751710000042
In the formula, R 2Be selected from C 1-C 6Alkyl, the C that halogen replaces 1-C 6Alkyl, the optional phenyl that replaces, C 1-C 3Alkyl-carbonyl, the optional benzoyl that replaces, carboxyl, formamyl, C 1-C 6Alkoxy carbonyl, hydroxyl, C 1-C 6Alkoxyl group, optional aryl or the heteroaryl that replaces, amino, the optional amino that replaces;
R 3Be selected from C 1-C 6Alkyl, the optional phenyl that replaces, C 1-C 3Alkyl-carbonyl, C 1-C 3Alkoxy carbonyl, the optional benzoyl that replaces, carboxyl, amide group, the optional aniline formyl radical that replaces;
R 4Be selected from C 5Above alkyl or cycloalkyl, the optional phenyl that replaces, 5 or 6 Yuans fragrant heterocyclic radicals.
In more preferred embodiment, R 2Be preferable over CH 3, phenyl; R 3Be preferable over C 1-C 3Alkoxyl formyl;
Herein, " alkyl " is often referred to saturated side chain and the straight chained alkyl that carbon chain lengths is 1-10 carbon atom, preferably a long 1-6 carbon atom, the more preferably alkyl of long 1-4 or 1-3 carbon atom." cycloalkyl " finger ring shape alkyl, it becomes the ring carbon atom number to be generally 3-8.Exemplary cycloalkyl comprises cyclopropyl, cyclobutyl, cyclopentyl, suberyl and cyclohexyl etc.
Herein, " alkenyl " refers to that straight or branched contains 2-10 carbon atom, wherein is the group that contains two keys between two carbon atoms in the chain at least.Preferred thiazolinyl is the thiazolinyl that contains 2-4 carbon atom.Typical alkenyl comprises vinyl, 1-propenyl, 2-methyl-1-propylene base, 1-butylene base and crotyl.
" alkynyl " used herein refers to that straight or branched contains 2-10 carbon atom, wherein is the group that contains three key between two carbon atoms in the chain at least.Preferred alkynyl is the alkynyl that contains 2-4 carbon atom.Typical alkynyl comprises ethynyl, 1-proyl, 1-methyl-2-propynyl, 2-propynyl, ethyl acetylene base and 2-butyne base.
Herein, " aryl " refers to contain monocycle, dicyclo or the three cyclophane family groups of 6 to 14 carbon atoms, comprises phenyl, naphthyl, phenanthryl, anthryl, indenyl, Fluorene base, tetrahydro naphthyl, indanyl etc.Aryl optionally is selected from following substituting group by 1-5 (for example, 1,2,3,4 or 5) and replaces: halogen, C 1-C 4Aldehyde radical, C 1-C 6The alkyl (for example trifluoromethyl) that replaces of straight or branched alkyl, cyano group, nitro, amino, hydroxyl, methylol, halogen, alkoxyl group (for example trifluoromethoxy), carboxyl, the C that halogen replaces 1-C 4Alkoxyl group, sulfydryl, C 1-C 10Alkylthio and C 1-C 4The benzyloxy of acyl group, morpholinyl, the optional aryl that replaces (for example optional phenyl that replaces), the optional aryloxy that replaces (for example optional phenoxy group that replaces) and optional replacement.For example, aryl can be selected from following group replacement by 1-3: fluorine, chlorine, bromine, C 1-C 4The phenyl of alkyl, trifluoromethyl, morpholinyl, methoxyl group, phenyl, methoxy substitution, phenoxy group, benzyloxy, benzyloxy, oxyethyl group and the nitro etc. that are replaced by halogen.
Term as used herein " heterocyclic radical " refers to ring structure single or that condense, can be aromatics or non-aromatic in nature, and it preferably contains 3-20 and becomes annular atoms, more preferably contain 5-14 annular atoms, wherein at least 1 and preferred at most can to 4 be the heteroatoms that is selected from O, S and N.Herein, the example of heterocyclic radical comprises furyl, thienyl, pyrryl, pyrrolidyl, imidazolyl, triazolyl, thiazolyl, tetrazyl, oxazolyl, isoxazolyl, pyrazolyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazinyl, quinolyl, isoquinolyl, quinoxalinyl, benzothiazolyl, benzoxazolyl, benzothienyl, benzofuryl, morpholinyl, carbazyl, dibenzothiophene and 1,2-methylenedioxyphenyl.Herein, heterocyclic radical is optionally replaced by 1-3 substituting group as herein described.
Term as used herein " heteroatoms " comprises O, S and N.When heteroatoms was N, this N atom can be further by for example hydrogen or C 1-C 10The group of alkyl replaces.
Term as used herein " heteroaryl " or " fragrant heterocyclic radical " refer to have as mentioned above those heterocyclic radicals of aromatic character, include but not limited to furyl, thienyl, pyrryl, pyridyl, oxazolyl, pyrazinyl, pyridazinyl, pyrimidyl etc.
Term as used herein " halogen " comprises fluorine, chlorine, bromine and iodine.
Except as otherwise noted, term as used herein " optional replacement " refers to that its group of modifying optionally is selected from following substituting group by 1-5 (being generally 1,2 or 3) and replaces: C 1-C 4Alkyl, carboxyl, halogen, C 1-C 4Alkoxyl group, cyano group, nitro, amino, hydroxyl, aldehyde radical, C 1-C 6The C that acyl group, methylol, halogen replace 1-C 4The C that alkyl (for example trifluoromethyl), halogen replace 1-C 4Alkoxyl group (for example trifluoromethoxy), sulfydryl and C 1-C 4Acyl group.
Herein, amide group self or as the part of other group refers to " C 1-C 6Alkyl-CO-NH-" group.Exemplary amide group includes but not limited to formamido-, acetamido etc.
Herein, acyl group self or as the part of other group can contain 1-6 carbon atom, preferred 1-3 carbon atom.Exemplary acyl group includes but not limited to formyl radical, ethanoyl etc.
The preferred hereinafter compound of the numbering 1-77 shown in the table 1 of the present invention, especially preferred inhibiting rate be at those compounds more than 50%, includes but not limited to compound 1,3,5-8,10,12-49,52-65 and 67 etc.The present invention is compound 20,21,34-36 and 44 particularly preferably.
Compound of the present invention can adopt following methods to prepare:
Figure BDA0000099751710000061
Compound 1-18
Figure BDA0000099751710000062
Compound 19-45
Figure BDA0000099751710000063
Compound 46-63
In the above-mentioned preparation flow, R 1Civilian described as defined above, R is the substituting group on the optional aryl that replaces, R 6Satisfy previously described R with its carbonyl that connects 3Definition.Other compound of the present invention can be according to above-mentioned flow process preparation.Those skilled in the art can prepare needs according to reality, and adopting the conventional various initial compounds that obtain in this area is raw material, prepares compound of the present invention.
Second aspect present invention comprises a kind of pharmaceutical composition, and said composition contains the formula I of the present invention, the II that treat significant quantity and/or compound or its pharmacy acceptable salt of III, and pharmaceutically acceptable carrier or vehicle.
The example of the pharmacy acceptable salt of the compounds of this invention includes but not limited to inorganic and organic acid salt, for example hydrochloride, hydrobromate, vitriol, Citrate trianion, lactic acid salt, tartrate, maleate, fumarate, mandelate and oxalate; And with alkali inorganic and organic alkali salt of forming of sodium hydroxyl, three (hydroxymethyl) amido methane (TRIS, amine trihydroxybutane) and N-METHYL-ALPHA-L-GLUCOSAMINE for example.
Although everyone demand is different, those skilled in the art can determine the optimal dose of every kind of activeconstituents in the pharmaceutical composition of the present invention.Generally speaking, compound of the present invention or its pharmacy acceptable salt, to Mammals oral administration every day, dose is according to about 0.0025 to 50 mg/kg of body weight.But preferably the per kilogram oral administration is about 0.01 to 10 milligram.For example, the unit oral dosage can comprise about 0.01 to 50 milligram, preferably about 0.1 to 10 milligram the compounds of this invention.Unitary dose can give one or many, and be one or more pieces every day, and every contains and has an appointment 0.1 to 50 milligram, eligibly about 0.25 to 10 milligram the compounds of this invention or its solvate.
Pharmaceutical composition of the present invention can be formulated into the dosage form that is fit to various route of administration, includes but not limited to be formulated into for outside the intestines, and is subcutaneous, vein, muscle, intraperitoneal, transdermal, oral cavity, in the sheath, encephalic, the form of nasal cavity or external application administration is used for the treatment of tumour and other diseases.Dosage is effectively to improve or eliminate the dose of one or more illnesss.For the treatment of specified disease, significant quantity is the dose that is enough to improve or alleviate in some mode the symptom relevant with disease.Such dose can be used as single dose and uses, perhaps can be according to effective treatment plan administration.Dosage is also permitted cure diseases, but administration is normally in order to improve the symptom of disease.Generally need repetitively administered to realize required doing well,improving.The dosage of medicine will be according to patient's age, health and body weight, and the kind of concurrent treatment, the frequency for the treatment of, and required treatment benefit decides.
Pharmaceutical preparation of the present invention can give any Mammals, as long as they can obtain the result for the treatment of of the compounds of this invention.Of paramount importance in these Mammalss is human.
Compound of the present invention or its drug regimen can be used for treating and the disease of preventing various DHODH mediations, especially relevant with the inhibition of DHODH disease.Herein, the disease of DHODH mediation mainly is various because the disease that certain class cell fast breeding causes, such as cancer, and inflammatory reaction and by host's rejection of the same race or that xenotransplant causes.The disease of DHODH mediation also comprises various autoimmune diseases.Specifically, the disease of DHODH mediation includes but not limited to rheumatoid arthritis, colitis, lupus erythematosus (comprising systemic lupus erythematous), renal glomerular disease (comprising multiple Secondary cases and primary glomerulopathy), anti-organ graft rejection, melanoma, psoriatic arthritis, psoriatic, mammary cancer, prostate cancer, squamous cell carcinoma of the head and neck and multiple marrow cancer etc.
Pharmaceutical preparation of the present invention can be made in a known manner.For example, by traditional mixing, granulate ingot processed, dissolving, or freezing dry process manufacturing.When making oral preparations, can be in conjunction with solid adjuvant material and active compound, the selectivity grinding mixture.If after need to or adding appropriate amount of addition agent in case of necessity, the processing granular mixture obtains tablet or lozenge core.
Suitable auxiliary material is filler particularly, for example carbohydrate such as lactose or sucrose, N.F,USP MANNITOL or sorbyl alcohol; Cellulose preparation or calcium phosphate, for example tricalcium phosphate or secondary calcium phosphate; And binding agent, for example starch paste comprises W-Gum, wheat starch, Starch rice, yam starch, gelatin, tragacanth, methylcellulose gum, Vltra tears, Xylo-Mucine, or polyvinylpyrrolidone.If necessary, can increase disintegrating agent, such as starch above-mentioned, and carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, or alginic acid or its salt, such as sodium alginate. auxiliary is flowing regulator and lubricant particularly, for example, silica, talcum, stearates, such as the magnesium calcium stearate, stearic acid or polyoxyethylene glycol.If necessary, Ke Yi Give lozenge nuclear core provides the suitable dressing that can resist gastric juice.For this reason, can use concentrated saccharide solution.This solution can contain Sudan Gum-arabic, talcum, polyvinylpyrrolidone, polyoxyethylene glycol and/or titanium dioxide, lacquer solution and suitable organic solvent or solvent mixture.In order to prepare the dressing of resistant to gastric juice, can use suitable cellulose solution, for example cellulose acetate phthalic acid or Vltra tears phthalic acid.Can add dyestuff or pigment to the dressing of tablet or lozenge nuclear core.For example, for identification or in order to characterize the combination of activeconstituents dosage.
Therefore, third aspect present invention provides a kind of method of disease of the DHODH for the treatment of mediation, and the method comprises that the object that needs is with compound of the present invention or pharmaceutical composition.
Medication includes but not limited to the various medications that this area is known, can be determined according to patient's practical situation.These methods include but not limited to outside the intestines, and are subcutaneous, vein, and muscle, intraperitoneal, transdermal, the oral cavity, in the sheath, encephalic, nasal cavity or external application administration.
The present invention also comprises the purposes of the compounds of this invention in the medicine that the disease of preparation treatment or prevention DHODH mediation is used.The present invention also comprises the purposes of the compounds of this invention in the medicine of the active usefulness of preparation inhibition DHODH.
Embodiment
In following embodiment, will further illustrate the present invention.These embodiment only are used for explanation the present invention, but do not limit the present invention in any way.
Composite part
Embodiment 1: the synthetic logical method of compound 1-18:
With chloro ethyl acetoacetate (9.52g, 57.84mmol) be dissolved in the 100mL ethanol, add thioacetamide (4.8g, 63.9mmol) under the agitation condition, be warming up to reflux temperature, stirring and refluxing 2h, question response finishes, and the reaction solution decompression is desolventized, and gets tangerine pink solid 9.80g, yield 91.1% is directly used in next step reaction.
Take by weighing LiOHH 2O (1.25g, 29.7mmol) is dissolved in the 50mL water, adds the ester (2.5g that single step reaction is synthetic under the agitation condition, 13.5mmol), be warming up to 90 ℃, along with reaction is carried out, solid dissolves gradually, and stopped reaction behind the 1h uses dilute hydrochloric acid conditioned reaction liquid pH to neutral slant acidity, there are a large amount of solids to separate out, suction filtration, after filter cake washes with water, drying, get faint yellow solid 2.05g, yield 96.6%.
Figure BDA0000099751710000091
Take by weighing thiazol formic-acid (1.1eq) and HATU (1.2eq) is dissolved among the 7mL DMF, after stirring, drip 8 DIPEA, stir 5min after, add again aromatic amine compounds (1eq), stir under the room temperature condition and spend the night, TLC follows the tracks of reaction.
Post-treating method one: in reaction solution, drip saturated ammonium chloride solution, solution becomes muddy a large amount of solids that occur gradually, drip saturated ammonium chloride solution to solid no longer occurring, the suction filtration reaction solution, filter cake is washed with saturated aqueous ammonium chloride first, wash with saturated sodium bicarbonate aqueous solution again, dry cake, namely get target compound, with ethyl acetate/saturated ammonium chloride it is extracted, the organic phase that collection obtains reduces pressure after with anhydrous sodium sulfate drying and desolventizes, and the solid that obtains is further purification by silica gel column chromatography (PE: EA=5: 1) again.
The target compound spectral data
N-(4-chloro-phenyl-)-2,4-dimethylthiazole-5-methane amide
Figure BDA0000099751710000092
1H?NMR(400MHz，DMSO)δ(ppm)：10.24(s，1H)，7.70(d，J＝8.8Hz，2H)，7.40(d，J＝8.8Hz，2H)，2.66(s，3H)，2.54(s，3H)。
HRMS(ESI)calcd?for?C 12H 11ClN 2OS(M+H)267.0359，found?267.0355。
N-[4-(trifluoromethyl) phenyl]-2,4-dimethyl-thiazole-5-methane amide
Figure BDA0000099751710000093
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.69(d，J＝8.4Hz，2H)，7.62(d，J＝8.8Hz，2H)，7.51(s，1H)，2.73(s，3H)，2.72(s，3H)。
HRMS(ESI)calcd?for：C 13H 13FN 2OS(M+H)265.0811，found?265.0805。
N-(3-fluoro-4-aminomethyl phenyl)-2,4-dimethylthiazole-5-methane amide
Figure BDA0000099751710000101
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.46(dd，J 1＝2.0Hz，J 2＝11.2Hz，1H)，7.41(s，1H)，7.15(t，J＝8.0Hz，1H)，7.10(dd，J 1＝2.0Hz，J 2＝8.4Hz，1H)，2.72(s，6H)，2.26(d，J＝1.6Hz，3H)。
HRMS(ESI)calcd?for：C 13H 13FN 2OS(M+H)265.0811，found?265.0805。
N-(4-bromo-2-aminomethyl phenyl)-2,4-dimethylthiazole-5-methane amide
Figure BDA0000099751710000102
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.80(d，J＝9.2Hz，1H)，7.37-7.35(m，2H)，7.21(s，1H)，2.73(s，3H)，2.71(s，3H)，2.28(s，3H)。
HRMS(ESI)calcd?for：C 13H 13BrN 2OS(M+H)325.0010，found?325.0009。
N-(9-ethyl-9H-carbazole-3-yl)-2,4-dimethylthiazole-5-methane amide
Figure BDA0000099751710000103
1H?NMR(400MHz，CDCl 3)δ(ppm)：8.36(s，1H)，8.10(d，J＝7.6Hz，1H)，7.57-7.55(m，2H)，7.52-7.49(m，1H)，7.44-7.39(m，2H)，7.25(t，J＝7.2Hz，1H)，4.41-4.36(m，2H)，2.78(s，3H)，2.74(s，3H)，1.45(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 20H 19N 3OS(M+H)350.1327，found?350.1332。
N-(anthracene-2-yl)-2,4-dimethylthiazole-5-methane amide
Figure BDA0000099751710000104
1H?NMR(400MHz，DMSO)δ(ppm)：8.52(s，2H)，8.48(s，1H)，8.08-8.04(m，3H)，7.70(dd，J 1＝1.6Hz，J 2＝9.2Hz，1H)，7.50-7.47(m，2H)，2.68(s，3H)，2.59(s，3H)。
HRMS(ESI)calcd?for：C 20H 16N 2OS(M-H)331.0905，found?331.0907。
N-(4-bromo-3-(trifluoromethyl) phenyl)-2,4-dimethylthiazole-5-methane amide
Figure BDA0000099751710000111
1H?NMR(400MHz，CDCl 3)δ(ppm)：8.76(d，J＝4.4Hz，1H)，8.47(d，J＝8.4Hz，1H)，7.50-7.47(m，1H)，2.82(s，3H)，2.80(s，3H)。
HRMS(ESI)calcd?for：C 13H 10BrF 3N 2OS(M-H)376.9571，found?376.9572。
N-(3,5-dichlorophenyl)-2,4-dimethylthiazole-5-methane amide
Figure BDA0000099751710000112
1H?NMR(400MHz，CDCl 3)δ(ppm)：8.77(dd，J 1＝1.2Hz，J 2＝4.4Hz，1H)，8.48(dd，J 1＝1.2Hz，J 2＝8Hz，1H)，7.51-7.48(m，1H)，2.83(s，3H)，2.81(s，3H)。
HRMS(ESI)calcd?for：C 12H 10C 12N 2OS(M-H)298.9813，found?298.9806。
2,4-dimethyl-N-(4-morpholinyl phenyl) thiazole-5-methane amide
Figure BDA0000099751710000113
1H?NMR(400MHz，DMSO)δ(ppm)：9.86(s，1H)，7.50(d，J＝8.4Hz，2H)，6.92(d，J＝8.8Hz，2H)，3.73(t，J＝4.4Hz，4H)，3.06(t，J＝4.4Hz，4H)，2.65(s，3H)，2.52(s，3H)。
HRMS(ESI)calcd?for：C 16H 19N 3O 2S(M+H)318.1276，found?318.1276。
2,4-dimethyl-N-phenyl thiazole-5-methane amide
Figure BDA0000099751710000114
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.57(d，J＝7.6Hz，2H)，7.43(s，1H)，7.39(t，J＝7.6Hz，2H)，7.18(t，J＝7.2Hz，1H)，2.75(s，3H)，2.74(s，3H)。
HRMS(ESI)calcd?for：C 12H 12N 2OS(M+H)233.0749，found?233.0745。
2,4-dimethyl-N-(naphthalene-2-yl) thiazole-5-methane amide
Figure BDA0000099751710000121
1H?NMR(400MHz，DMSO)δ(ppm)：10.28(s，1H)，8.33(d，J＝2.4Hz，1H)，7.90-7.84(m，3H)，7.72(dd，J 1＝2.0Hz，J 2＝10.0Hz，1H)，7.49-7.42(m，2H)，2.68(s，3H)，2.59(s，3H)。
HRMS(ESI)calcd?for：C 16H 14N 2OS(M+H)283.0905，found?283.0900。
N-(4-chloro-phenyl-)-4-methylthiazol-5-methane amide
Figure BDA0000099751710000122
1H?NMR(400MHz，CDCl 3)δ(ppm)：8.81(s，1H)，7.55(d，J＝8.8Hz，2H)，7.52(s，1H)，7.36(d，J＝8.8Hz，2H)，2.83(s，3H)。
HRMS(ESI)calcd?for：C 11H 9ClN 2OS(M+H)253.0202，found?253.0206。
N-(anthracene-2-yl)-4-methylthiazol-5-methane amide
Figure BDA0000099751710000123
1H?NMR(400MHz，DMSO)δ(ppm)：9.17(s，1H)，8.53(d，J＝4.4Hz，2H)，8.50(s，1H)，8.09-8.05(m，3H)，7.72(dd，J 1＝2.0Hz，J 2＝10.0Hz，1H)，7.51-7.46(m，2H)，2.68(s，3H)。
HRMS(ESI)calcd?for：C 19H 14N 2OS(M-H)317.0749，found?317.0750。
N-(9-ethyl-9H-carbazole-3-yl)-4-methylthiazol-5-methane amide
Figure BDA0000099751710000124
1H?NMR(400MHz，DMSO)δ(ppm)：10.25(s，1H)，9.14(s，1H)，8.45(s，1H)，8.10(d，J＝8.0Hz，1H)，7.68-7.66(m，1H)，7.61(s，1H)，7.61-7.59(m，1H)，7.46(t，J＝7.6Hz，1H)，7.19(t，J＝7.6Hz，1H)，4.47-4.42(m，2H)，2.66(s，3H)，1.31(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 19H 17N 3OS(M+H)336.1171，found?336.1176。
4-methyl-N-(naphthalene-2-yl) thiazole-5-methane amide
Figure BDA0000099751710000131
1H?NMR(400MHz，DMSO)δ(ppm)：10.43(s，1H)，9.15(s，1H)，8.35(s，1H)，7.91-7.85(m，3H)，7.73(d，J＝8.8Hz，1H)，7.52-7.42(m，2H)，2.66(s，3H)。
HRMS(ESI)calcd?for：C 15H 12N 2OS(M-H)267.0592，found?267.0590。
4-methyl-N-phenyl thiazole-5-methane amide
Figure BDA0000099751710000132
1H?NMR(400MHz，CDCl 3)δ(ppm)：8.77(s，1H)，7.63(s，1H)，7.58(d，J＝7.6Hz，2H)，7.38(t，J＝8.0Hz，2H)，7.19(t，J＝8.4Hz，1H)，2.80(s，3H)。
HRMS(ESI)calcd?for：C 11H 10N 2OS(M-H)217.0436，found?217.0435。
N-(dibenzo (b, d) thiophene-2-yl)-2,4-dimethylthiazole-5-methane amide
Figure BDA0000099751710000133
HRMS(ESI)calcd?for：C 18H 14N 2OS 2(M+H)339.0626，found?339.0620。
N-(dibenzo (b, d) thiophene-2-yl)-2-methylthiazol-5-methane amide
Figure BDA0000099751710000134
HRMS(ESI)calcd?for：C 17H 12N 2OS 2(M+H)325.0469，found?325.0468。
Embodiment 2: the synthetic logical method of compound 19-45:
Synthesizing of thiocarbamide
Arylamine (8mmol, 1eq) is dissolved in the 24mL acetone, under agitation condition, adds the triethylene diamine (24mmol that weighs up, 3eq), drip again dithiocarbonic anhydride 20mL, a large amount of solids occur, continue stirring at room 24h, the suction filtration reaction solution, filter cake is washed with sherwood oil, after the drying, filter cake is dissolved in the 50mL chloroform, takes by weighing BTC (2.7mmol, 0.33eq) and be dissolved in the 30mL chloroform, drop in the 1h in the reaction solution, stir this solution under the room temperature and spend the night.After reaction finishes, the suction filtration reaction solution, filter cake is washed with methylene dichloride, and the filtrate that obtains directly adds silica gel and is spin-dried for rear dry method loading (PE wash-out).
NCS is dissolved in a small amount of methylene dichloride, strengthens large excessive ammoniacal liquor, 0 ℃ of lower stirring 3h, suction filtration reaction solution, filter cake wash with water, and dry cake namely gets thiocarbamide, is directly used in next step reaction.
Synthesizing of target compound
Figure BDA0000099751710000142
Compound 19-45
Beta-cyclodextrin (590mg, 0.52mmol) is dissolved in 10mL H 2Among the O, this solution is warming up to 50 ℃ all to be dissolved to beta-cyclodextrin and to form a water white solution, ethyl benzoylacetate is dropped in the beta-cyclodextrin aqueous solution after with the 0.5mL acetone diluted, take by weighing NBS (138.9mg after stirring, 0.780mmol) add this mixing solutions, after stirring 1h, add again fragrant thiocarbamide (0.52mmol), TLC follows the tracks of reaction, question response carries out fully, with ethyl acetate/saturated aqueous common salt extractive reaction liquid, to collect the organic phase obtain and reduce pressure after with anhydrous sodium sulfate drying and desolventize, the crude product that obtains is further purified (PE: EA=10: 1) with silica gel column chromatography.
The target compound spectral data
2-anilino-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000143
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.74-7.72(m，2H)，7.38(d，J＝3.6Hz，3H)，7.38(t，J＝8.0Hz，2H)，7.14(d，J＝6.8Hz，3H)，4.23(q，J＝7.2Hz，2H)，1.26(t，J＝6.8Hz，3H)。
HRMS(ESI)calcd?for：C 18H 16N 2O 2S(M+H)325.1011，found?325.1009。
2-(3,4-dimethyl) anilino-4-phenyl-5-thiazole ethyl formate
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.74-7.72(m，2H)，7.40-7.39(m，3H)，7.12(d，J＝8.4Hz，1H)，7.02(dd，J 1＝2.4Hz，J 2＝8.0Hz，1H)，6.97(d，J＝2.0Hz，1H)，4.23(q，J＝7.2Hz，2H)，2.27(s，6H)，1.26(t，J＝6.8Hz，3H)。
HRMS(ESI)calcd?for：C 20H 20N 2O 2S(M+H)353.1324，found?353.1324。
2-(3-chloro-4-methyl)-anilino-4-phenyl-5-thiazole ethyl formate
1H?NMR(400MHz，DMSO)δ(ppm)：7.82(d，J＝2.0Hz，1H)，7.74-7.72(m，2H)，7.45-7.42(m，4H)，7.32(d，J＝8.4Hz，1H)，4.16(q，J＝7.2Hz，2H)，2.28(s，3H)，1.18(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 19H 17ClN 2O 2S(M+H)373.0778，found?373.0778。
2-(4-methoxyl group)-anilino-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000153
1H?NMR(400MHz，DMSO)δ(ppm)：10.59(s，1H)，7.73-7.70(m，2H)，7.52(d，J＝9.2Hz，2H)，7.42(t，J＝3.2Hz，3H)，6.96(dd，J 1＝2.0Hz，J 2＝6.8Hz，2H)，4.13(q，J＝7.2Hz，2H)，3.74(s，3H)，1.16(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 19H 18N 2O 3S(M+H)355.1116，found?355.1115。
2-(3-trifluoromethyl-4-bromine)-anilino-4-phenyl-5-thiazole ethyl formate
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.69-7.67(m，2H)，7.53(d，J＝8.8Hz，1H)，7.47(d，J＝2.8Hz，1H)，7.33-7.29(m，3H)，7.10(dd，J 1＝2.8Hz，J 2＝8.6Hz，1H)，4.26(q，J＝7.2Hz，2H)，1.28(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 19H 14BrF 3N 2O 2S(M+H)470.9990，found?470.9986。
2-(3,4-dichloro)-anilino-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000161
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.70-7.67(m，2H)，7.35(dd，J 1＝2.0Hz，J 2＝5.4Hz，3H)，7.26(d，J＝8.8Hz，1H)，7.14(d，J＝2.4Hz，1H)，6.88(dd，J 1＝2.8Hz，J 2＝8.6Hz，1H)，4.24(q，J＝7.2Hz，2H)，1.27(t，J＝7.2Hz，3H)
2-(4-chlorine)-anilino-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000162
1H?NMR(400MHz，CDCl 3)δ(ppm)：10.59(s，1H)，7.67(d，J＝6.4Hz，2H)，7.36-7.28(m，3H)，7.13(d，J＝8.4Hz，2H)，6.85(d，J＝8.8Hz，2H)，4.23(q，J＝7.2Hz，2H)，1.25(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 18H 15ClN 2O 2S(M+H)359.0621，found?359.0615。
2-(2-methyl-4-bromine)-anilino-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000163
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.52-7.50(m，2H)，7.36(dd，J 1＝2.4Hz，J 2＝8.4Hz，1H)，7.29(d，J＝2.0Hz，1H)，7.27-7.23(m，2H)，7.15(t，J＝3.6Hz，2H)，4.17(q，J＝7.2Hz，2H)，2.08(s，3H)，1.21(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 19H 17BrN 2O 2S(M+H)417.0272，found?417.0271。
2-anthracene amino-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000164
1H?NMR(400MHz，CDCl 3)δ(ppm)：8.43(s，2H)，8.03(d，J＝8.8Hz，3H)，7.99(d，J＝2.0Hz，1H)，7.83-7.81(m，2H)，7.54-7.46(m，5H)，7.27(d，J＝2.0Hz，1H)，4.30(q，J＝7.2Hz，2H)，1.31(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 26H 20N 2O 2S(M+H)425.1324，found?425.1324。
2-(3-trifluoromethyl-4-chlorine)-anilino-4-phenyl-5-thiazole ethyl formate
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.70-7.67(m，2H)，7.50(d，J＝2.8Hz，1H)，7.38-7.32(m，4H)，7.22(dd，J 1＝2.8Hz，J 2＝8.8Hz，1H)，4.26(q，J＝7.2Hz，2H)，1.27(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 19H 14ClF 3N 2O 2S(M+H)427.0495，found?427.0493。
2-(4-trifluoromethyl)-anilino-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000172
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.78-7.75(m，2H)，7.59(d，J＝8.4Hz，2H)，7.43-7.41(m，3H)，7.34(d，J＝8.4Hz，2H)，4.27(q，J＝7.2Hz，2H)，1.29(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 19H 15F 3N 2O 2S(M+H)393.0885，found?393.0884。
2-((3-fluoro-[1,1 '-biphenyl]-4-yl) amino)-4-phenyl-5-thiazole ethyl formate
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.98(t，J＝8.4Hz，1H)，7.81-7.79(m，2H)，7.61-7.59(m，2H)，7.50-7.40(m，8H)，4.21(q，J＝7.2Hz，2H)，1.30(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 24H 19FN 2O 2S(M+H)419.1230，found?419.1232。
2-(benzo [d] [1,3] dioxolane-5-base-amino)-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000174
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.68-7.66(m，2H)，7.37-7.34(m，3H)，6.75(d，J＝8.0Hz，1H)，6.65-6.68(m，2H)，6.01(s，2H)，4.20(q，J＝7.2Hz，2H)，1.24(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 19H 16N 2O 4S(M+H)369.0909，found?369.0902。
2-((9-ethyl-9H-carbazole-3-yl) amino)-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000181
1H?NMR(400MHz，DMSO)δ(ppm)：10.71(s，1H)，8.36(d，J＝1.2Hz，1H)，8.12(d，J＝7.6Hz，1H)，7.79-7.76(m，2H)，7.67-7.61(m，3H)，7.49-7.44(m，4H)，7.21(t，J＝7.2Hz，1H)，4.48-4.43(m，2H)，4.13(q，J＝7.2Hz，2H)，1.33(t，J＝7.2Hz，3H)，1.12(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 26H 23N 3O 2S(M+H)442.1589，found?442.1582。
2-(3,5 dichloro)-anilino-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000182
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.74-7.72(m，2H)，7.39-7.37(m，3H)，7.06(d，J＝1.6Hz，1H)，7.00(d，J＝1.6Hz，2H)，4.24(q，J＝7.2Hz，2H)，，1.29(t，J＝7.2Hz，3H)。
2-((5,6,7,8-naphthane-2-yl) amino)-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000183
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.76-7.73(m，2H)，7.42-7.41(m，3H)，7.72(d，J＝8.4Hz，1H)，7.02(dd，J 1＝2.4Hz，J 2＝7.4Hz，1H)，6.94(d，J＝2.0Hz，1H)，4.24(q，J＝7.2Hz，2H)，2.78(s，4H)，1.85-1.81(m，4H)，1.26(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 22H 22N 2O 2S(M+H)379.1480，found?379.1481
2-(the 4-tertiary butyl)-phenylamino-4-phenyl-5-thiazole ethyl formate
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.75-7.73(m，2H)，7.40-7.38(m，5H)，7.19(d，J＝8.4Hz，2H)，4.24(q，J＝7.2Hz，2H)，1.36(s，9H)，1.27(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 22H 24N 2O 2S(M+H)381.1637，found?381.1635
2-(3-fluoro-4-methyl)-anilino-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000192
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.72-7.70(m，2H)，7.38-7.37(m，3H)，7.08(t，J＝7.6Hz，1H)，6.84-6.78(m，2H)，4.23(q，J＝7.2Hz，2H)，2.25(s，3H)，1.26(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 19H 17FN 2O 2S(M+H)357.1073，found?357.1068
2-([1,1 '-biphenyl]-4-base-amino)-4-phenyl-5-thiazole ethyl formate
1H?NMR(400MHz，CDCl 3)δ(ppm)：8.11-8.09(m，1H)，7.75(dd，J 1＝2.4Hz，J 2＝6.6Hz，2H)，7.64-7.61(m，4H)，7.48-7.44(m，5H)，7.38(d，J＝8.0Hz，2H)，4.26(q，J＝7.2Hz，2H)，1.29(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 24H 20N 2O 2S(M+H)401.1324，found?401.1318。
2-((3 '-methoxyl group-[1,1 '-biphenyl]-4-yl) amino)-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000194
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.76-7.74(m，2H)，7.51(d，J＝8.4Hz，2H)，7.41-7.37(m，4H)，7.17(d，J＝8.4Hz，3H)，7.12(d，J＝2.0Hz，1H)，6.93(dd，J 1＝2.4Hz，J 2＝8.0Hz，1H)，4.25(q，J＝7.2Hz，2H)，(m，2H)，3.91(s，3H)，1.28(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 25H 22N 2O 3S(M+H)431.1429，found?431.1432
2-((3-fluoro-3 '-methoxyl group-[1,1 '-biphenyl]-4-yl) amino)-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000201
1H?NMR(400MHz，CDCl 3)δ(ppm)：8.00(t，J＝8.4Hz，1H)，7.82-7.79(m，2H)，7.48-7.38(m，6H)，7.18(d，J＝7.6Hz，1H)，7.12(s，1H)，6.94(dd，J 1＝2.4Hz，J 2＝8.4Hz，1H)，4.28(q，J＝7.2Hz，2H)，3.90(s，3H)，1.30(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 25H 21FN 2O 3S(M+H)449.1335，found?449.1331
2-((2,3-dihydro-1H-indenes-5-yl) amino)-4-phenyl-5-thiazole ethyl formate
Figure BDA0000099751710000202
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.74-7.73(m，2H)，7.39-7.38(m，3H)，7.19(d，J＝8.0Hz，1H)，7.07-6.99(m，2H)，4.22(q，J＝7.2Hz，2H)，2.91(t，J＝7.6Hz，4H)，2.14-2.07(m，2H)，1.26(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 21H 20N 2O 2S(M+H)365.1324，found?365.1326。
2-((3,5-, two fluoro-3 '-methoxyl group-[1,1 '-biphenyl]-4-yl) amino)-4-phenyl-5-thiazole ethyl formate
1H?NMR(400MHz，CDCl 3)δ(ppm)：7.74-7.73(m，2H)，7.39-7.38(m，3H)，7.19(d，J＝8.0Hz，1H)，7.07-6.99(m，2H)，4.19(q，J＝7.2Hz，2H)，2.91(t，J＝7.6Hz，4H)，2.14-2.07(m，2H)，1.26(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for：C 25H 20F 2N 2O 3S(M+H)467.1241，found?467.1235
Embodiment 3: the synthetic logical method of compound 46-63:
Figure BDA0000099751710000204
Compound 46-63
Synthetic method: get the substituted-phenyl thiocarbamide of 1mmol and the 2-chloracetyl acetone (2-chloroacetyl acetacetic ester) of 1mmol and be dissolved in 20mL methyl alcohol, backflow is spent the night, be spin-dried for methyl alcohol, add a small amount of rare wet chemical neutralization, the saturated common salt water washing, ethyl acetate extraction, organic layer concentrate drying column chromatography gets product.
The target compound spectral data
2-(4-methyl-3-chloroanilino)-4-methyl-5-acetylthiazole
Figure BDA0000099751710000211
1H?NMR(400MHz，DMSO)δ(ppm)：7.79(d，J＝2.0Hz，1H)，7.48-7.38(m，1H)，7.30(d，J＝8.0Hz，1H)，2.55(s，3H)，2.42(s，3H)，2.27(s，3H)。
HRMS(ESI)calcd?for?C 13H 13ClN 2OS(M+H +)281.0515，found?281.0515；
2-(3,4-toluidine)-4-methyl-5-acetylthiazole
Figure BDA0000099751710000212
1H?NMR(400MHz，DMSO)δ(ppm)：7.32(d，J＝8.0Hz，1H)，7.28(s，1H)，7.11(d，J＝8.4Hz，1H)，2.53(s，3H)，2.41(s，3H)，2.21(s，3H)，2.18(s，3H)。
HRMS(ESI)calcd?for?C 14H 16N 2OS(M+H +)261.1062，found?261.1065；
2-(4-bromo-3-trifluoromethylbenzene amido)-4-methyl-5-acetylthiazole
Figure BDA0000099751710000213
1H?NMR(400MHz，DMSO)δ(ppm)：8.25(s，1H)，7.82(s，3H)，2.57(s，3H)，2.45(s，3H)。
HRMS(ESI)calcd?for?C 13H 10BrF 3N 2OS(M+H +)378.9728，found?378.9724；
2-(2-naphthylamine base)-4-methyl-5-acetylthiazole)
Figure BDA0000099751710000214
1H?NMR(400MHz，DMSO)δ(ppm)：8.25(d，J＝2.0Hz，1H)，7.91(d，J＝8.8Hz，1H)，7.86(d，J＝8.8Hz，2H)，7.61(dd，J 1＝2.4Hz，J 2＝8.8Hz，1H)，7.51-7.47(m，1H)，7.43-7.39(m，1H)，2.62(s，3H)，2.46(s，3H)。
HRMS(ESI)calcd?for?C 16H 14N 2OS(M+H +)283.0905，found?283.0903；
2-(6-anthranylamine base)-4-methyl-5-acetylthiazole
Figure BDA0000099751710000215
1H?NMR(400MHz，DMSO)δ(ppm)：8.51-8.47(m，3H)，8.10-8.04(m，3H)，7.57-7.45(m，3H)，2.65(s，3H)，2.48(s，3H)。
HRMS(ESI)calcd?for?C 20H 16N 2OS(M+H +)333.1062，found?333.1057；
2-(4-chloro-3-trifluoromethylbenzene amido)-4-methyl-5-acetylthiazole
Figure BDA0000099751710000221
1H?NMR(400MHz，DMSO)δ(ppm)：11.13(s，1H)，8.26(d，J＝2.4Hz，1H)，7.91(dd，J 1＝2.4Hz，J 2＝8.8Hz，1H)，7.69(d，J＝8.8Hz，1H)，2.58(s，3H)，2.46(s，3H)。
HRMS (ESI) m/z: vacant (M+H +).
2-(4-phenetole amido)-4-methyl-5-acetylthiazole
Figure BDA0000099751710000222
1H?NMR(400MHz，DMSO)δ(ppm)：7.40(d，J＝8.4Hz，2H)，7.91(d，J＝8.8Hz，2H)，4.02-3.97(m，2H)，2.50(s，3H)，2.36(s，3H)，1.32(t，J＝6.8Hz，3H)。
HRMS(ESI)calcd?for?C 14H 16N 2O 2S(M+H +)277.1011，found?277.1009；
2-(9-ethyl-9H-carbazole-3-is amino)-4-methyl-5-acetylthiazole
Figure BDA0000099751710000223
1H?NMR(400MHz，DMSO)δ(ppm)：10.68(s，1H)，8.33(s，1H)，8.13(d，J＝8.0Hz，1H)，7.65-7.55(m，3H)，7.47(t，J＝7.2Hz，1H)，7.20(t，J＝7.2Hz，1H)，4.47-4.42(m，2H)，2.57(s，3H)，2.40(s，3H)，1.32(t，J＝6.0Hz，3H)。
HRMS(ESI)calcd?for?C 20H 19N 3OS(M+H +)350.1327，found?350.1319；
2-(anilino)-4-methyl-5-acetylthiazole
1H?NMR(400MHz，DMSO)δ(ppm)：7.60(d，J＝8.0Hz，2H)，7.37(t，J＝8.0Hz，2H)，7.05(d，J＝7.2Hz，1H)，2.56(s，3H)，2.43(s，3H)。
HRMS(ESI)calcd?for?C 12H 12N 2OS(M+H +)233.0749，found?233.0746；
2-(4-bromobenzene amido)-4-methyl-5-acetylthiazole
Figure BDA0000099751710000231
1H?NMR(400MHz，DMSO)δ(ppm)：7.61(d，J＝8.8Hz，2H)，7.53(d，J＝8.8Hz，2H)，2.56(s，3H)，2.44(s，3H)。
HRMS(ESI)calcd?for?C 12H 11BrN 2OS(M+H +)310.9854，found?310.9854；
2-(4-tert.-butylbenzene amido)-4-methyl-5-acetylthiazole
Figure BDA0000099751710000232
1H?NMR(400MHz，DMSO)δ(ppm)：7.49(d，J＝8.4Hz，2H)，7.38(d，J＝8.8Hz，2H)，2.54(s，3H)，2.42(s，3H)，1.28(s，9H)。
HRMS(ESI)calcd?for?C 16H 20N 2OS(M+H +)289.1375，found?289.1374；
2-(4-phenetole amido)-4-methyl-5-ethyl formate thiazole
1H?NMR(400MHz，DMSO)δ(ppm)：7.45(d，J＝9.2Hz，2H)，6.92(d，J＝8.8Hz，2H)，4.21-4.16(m，2H)，4.02-3.97(m，2H)，2.46(s，3H)，1.32(t，J＝6.8Hz，3H)，1.25(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for?C 15H 18N 2O 3S(M+H +)307.1116，found?307.1114；
2-(3,4-toluidine)-4-methyl-5-ethyl formate thiazole
1H?NMR(400MHz，DMSO)δ(ppm)：7.33(dd，J 1＝2.0Hz，J 2＝8.0Hz，1H)，7.28(s，1H)，7.11(d，J＝8.0Hz，1H)，4.23-4.17(m，2H)，2.22(s，3H)，2.18(s，3H)，1.26(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for?C 15H 18N 2O 2S(M+H +)291.1167，found?291.1169；
2-((9-ethyl-9H-carbazole-3-is amino)-4-methyl-5-ethyl formate thiazole
Figure BDA0000099751710000235
1H?NMR(400MHz，DMSO)δ(ppm)：10.57(s，1H)，8.32(d，J＝1.6Hz，1H)，8.13(d，J＝7.6Hz，1H)，7.65-7.55(m，3H)，7.47(d，J＝7.6Hz，1H)，7.20(d，J＝7.6Hz，1H)，4.47-4.42(m，2H)，4.22-4.16(m，2H)，2.54(s，3H)，1.33(t，J＝7.2Hz，3H)，1.25(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for?C 21H 21N 3O 2S(M+H +)380.1433，found?380.1436；
2-(2-anthranylamine base)-4-methyl-5-ethyl formate thiazole
Figure BDA0000099751710000241
1H?NMR(400MHz，DMSO)δ(ppm)：8.51-8.46(m，3H)，8.10-8.05(m，3H)，7.56-7.46(m，3H)，4.28-4.23(m，2H)，2.62(s，3H)，1.91(s，3H)，1.30(t，J＝7.2Hz，3H)。
2-(4-methyl 3-chloroanilino)-4-methyl-5-ethyl formate thiazole
Figure BDA0000099751710000242
1H?NMR(400MHz，DMSO)δ(ppm)：7.82(d，J＝2.0Hz，2H)，7.42-7.39(m，2H)，7.31(d，J＝8.4Hz，1H)，4.24-4.19(m，2H)，2.53(s，3H)，2.28(s，3H)，1.27(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for?C 14H 15ClN 2O 2S(M+H +)311.0621，found?311.0620；
2-(4-bromobenzene amido)-4-methyl-5-ethyl formate thiazole
Figure BDA0000099751710000243
1H?NMR(400MHz，DMSO)δ(ppm)：7.61(d，J＝8.8Hz，2H)，7.52(d，J＝8.8Hz，2H)，4.24-4.19(m，2H)，2.53(s，3H)，1.27(t，J＝7.2Hz，3H)。
HRMS(ESI)calcd?for?C 13H 13BrN 2O 2S(M+H +)340.9959，found?340.9951；
DHODH active testing part
Embodiment 1
Compound provided by the invention is to the active extracorporeal extracorporeal suppression of dihydroorate dehydrogenase (DHODH)
Material: the plasmid of people DHODH full length gene by Prof. Jon Clardy (Harvard Medical School) be so kind as to give (J.Bio.Chem.2008,283 (50), 35078-35085) or can buy from Aureal Dongyuan County bio tech ltd.The pET-19b carrier, E.coli DH5 α and E.coli BL21 (DE3) bacterial strain are all available from Novagen company.Restriction enzyme NdeI and Bam HI are available from NEB company.Primer is synthetic by Shanghai Ying Jun Bioisystech Co., Ltd.Other reagent is all available from Sigma.
According to people DHODH gene order design primer among the GenBank, forward primer is Fw:5 '-TGAACTACATATGGCCACGGGAGATGAG-3 '; Reverse primer Rv:5 '-ATATGGATCCTCACCTCCGATGATCTGC-3 '.Take the plasmid that contains the DHODH gene as template amplification, amplification condition: 95 ℃ of denaturation 2min; 94 ℃ of sex change 30s; 60 ℃ of annealing 45s; 72 ℃ are extended 1min 50s; 29 circulations, 72 ℃ are extended 10min fully.Reaction is carried out 1% agarose gel electrophoresis after finishing, and take 2000bp DNA standard as the relative molecular weight reference, checking amplified production relative molecular weight size is cut glue, reclaims amplified production with test kit.
Respectively PCR product and carrier pET-19b are carried out double digestion with Nde I and Bam H1, reclaim the goal gene and the carrier segments that digested, spend the night with 16 ℃ of connections of T4DNA ligase enzyme, make up recombinant expression vector pET-19b-DHODH.Recombinant plasmid transformed E.coli DH5a competence is inoculated on the LB flat board that contains ammonia benzyl mould and cultivates, and the positive bacterium colony of picking at random is inoculated in respectively in the little centrifuge tube that 2mL contains ammonia benzyl mould and spreads cultivation.Get bacterium liquid after spreading cultivation and extract in a small amount plasmid and carry out enzyme and cut and identify and bacterium liquid PCR evaluation, and carry out determined dna sequence by Shanghai Ying Jun Bioisystech Co., Ltd.
Recombinant plasmid pET-19b-DHODH Transformed E .coli BL21 (DE3) competence that order-checking is correct, coat on the LB flat board that contains penbritin and cultivate, the picking mono-clonal is inoculated in 37 ℃, 230rpm shaking table overnight incubation in the LB substratum that contains 100 μ M penbritins.Be inoculated in 37 ℃, 230rpm enlarged culturing in the LB substratum that 500mL contains 100 μ M penbritins in 1: 200 ratio.When treating that thalline OD value reaches 0.8-1, add IPTG in substratum, making the IPTG final concentration is 0.5mM.25 ℃ of abduction deliverings that spend the night.4 ℃, the centrifugal collection of 4000rpm are induced rear thalline, collect bacterial sediment with recentrifuge behind the deionized water wash, are put in-80 ℃ of preservations.
Use the resuspended thalline of lysate during protein purification.Lysate contains 50mM HEPES (pH 8.0), 0.15MNaCl, and the 10mM imidazoles, 10% glycerine, 0.1%Triton X-100 adds a little STI, ultrasonication thalline behind the resuspended mixing.4 ℃ of broken liquid, the centrifugal 30min of 10000rpm.Getting cleer and peaceful precipitation carries out protein electrophoresis and determines the albumen existence form.Supernatant is added combination in the ready Ni-NTA chromatography column, collect and pass liquid.Wash resin 3-5 time with the lysate that contains the 20mM imidazoles again, at last with the lysate eluted protein that contains the 300mM imidazoles, protein liquid behind the collection wash-out.Get the above 10 μ L albumen samples that respectively go on foot and carry out the SDS-PAGE electrophoresis, the testing goal protein content.Protein liquid is dialysed in dialyzate and is gone imidazoles, dialyzate to contain 50mM HEPES (pH 8.0), 0.15MNaCl, 10% glycerine, 0.1%Triton X-100 behind the wash-out.
DHODH is active to be determined by the method for measuring the DCIP minimizing, under the condition that coenzyme Q-10 exists, and DHODH catalytic substrate DHO, and two H are transferred on the prothetic group FMN of DHODH, and then pass to coenzyme Q-10, and pass to DCIP by coenzyme Q-10 at last, DCIP is reduced.Determine that by the amount of measuring the DCIP that per minute reduces enzyme lives.Measuring method adopts 96 orifice plates to be read by the BioTek microplate reader.Every hole contains surveys the liquid 199 μ L (50mMHEPES (pH 8.0), 0.15M KCl, 100 μ M coenzyme Q-10s, 100 μ M DCIP) that live.Add 0.2 μ M inhibitor (being dissolved in DMSO), making the inhibitor final concentration is 10 μ M, and incubated at room 10min adds 1 μ L substrate DHO at last, and making the DHO final concentration is 500 μ M.Read 6min with microplate reader under the 600nm wavelength behind the mixing, interval 30s reads a secondary data.Each the experiment establish at least 3 parallel.Inhibiting rate by the compound different concns calculates half effective inhibition concentration (IC at last 50).
Prepare as stated above other compound in the table, and tested its activity.The result is presented at following table 1:
Table 1: compound structure and hDHODH inhibiting rate
Figure BDA0000099751710000261
Figure BDA0000099751710000291
Figure BDA0000099751710000301
Figure BDA0000099751710000311
Figure BDA0000099751710000321
Figure BDA0000099751710000331
Figure BDA0000099751710000341

Claims (10)

1. following formula I compound:
Figure FDA0000099751700000011
In the formula,
R 1Be selected from H, C 1-C 6Alkyl, C 2-C 6Alkenyl or alkynyl, the optional aryl that replaces, nitro, amino, NR 4R 5, halogen;
R 2Be selected from H, C 1-C 6Alkyl, the C that halogen replaces 1-C 6Alkyl, C 1-C 6Unsaturated alkyl, C 1-C 3Alkyl-carbonyl, the optional benzoyl that replaces, carboxyl, aminocarboxyl, C 1-C 6Alkoxy carbonyl, hydroxyl, C 1-C 6Alkoxyl group, the optional aryl that replaces, the optional heteroaryl that replaces, amino, NHR 6
R 3Be selected from H, C 1-C 6Alkyl ,-C (O) NHR 7, C 1-C 6Alkoxy carbonyl, the alkyl that halogen replaces, C 2-C 6Alkenyl or alkynyl, the optional phenyl that replaces, C 1-C 4Alkyl-carbonyl, the optional benzoyl that replaces, the optional pyridyl carbonyl that replaces, C 1-C 3Alkyl carboxyl, amide group, amino, C 1-C 10Amino and halogen that alkyl replaces;
R 4And R 5Independently be selected from separately H, the optional aryl that replaces, the optional heterocyclic radical that replaces, the optional aryl carbonyl that replaces, the optional heterocyclic radical carbonyl that replaces, and the optional aryloxy alkyl carbonyl that replaces;
R 6Be selected from C 1-C 6Alkyl, the optional phenyl that replaces, or form 6 Yuans rings with the N that is connected, such as piperidine ring, or 6 Yuans contain oxygen or nitrogen heterocyclic, such as piperazine ring and morpholine ring; With
R 7Be selected from the optional aryl that replaces and the optional heterocyclic radical that replaces.
2. compound as claimed in claim 1 is characterized in that, described compound is selected from the compound of Formula Il:
Ar is selected from the optional aryl that replaces, the optional heterocyclic radical that replaces, the optional aryl carbonyl that replaces, the optional heterocyclic radical carbonyl that replaces, and the optional aryloxy alkyl carbonyl that replaces;
R 2Be selected from C 1-C 6Alkyl, the C that halogen replaces 1-C 6Alkyl, C 1-C 3Alkyl-carbonyl, the optional benzoyl that replaces, carboxyl, aminocarboxyl, C 1-C 6Alkoxy carbonyl, hydroxyl, C 1-C 6Alkoxyl group, optional aryl or the heteroaryl that replaces, amino and NHR 6
R 3Be selected from C 1-C 6Alkyl, C 1-C 6Alkoxy carbonyl, the optional phenyl that replaces, C 1-C 4Alkyl-carbonyl, the optional benzoyl that replaces, C 1-C 3Alkyl carboxyl, amide group and the optional aniline formyl radical that replaces.
3. compound as claimed in claim 2, the substituting group on the Ar is selected from C 1-C 10Alkyl, C 3-C 8Cycloalkyl, C 1-C 4Alkoxyl group, the optional phenyl that replaces, the optional phenoxy group that replaces, benzyloxy, CF 3, and halogen, substituent quantity is 1,2,3,4 or 5.
4. compound as claimed in claim 2 is characterized in that, R 2Be selected from C 1-C 6Alkyl, CF 3, phenyl, ethanoyl, benzoyl, carboxyl, formamyl, C 1-C 6Alkoxy carbonyl, amino.Methyl more preferably, CF 3, phenyl.
5. compound as claimed in claim 2 is characterized in that, R 3Be selected from C 1-C 6Alkyl, phenyl, C 1-C 3Alkyl-carbonyl, the optional benzoyl that replaces, carboxyl, C 1-C 6Alkoxy carbonyl, amide group, the optional aniline formyl radical that replaces; Preferred C 1-C 3Alkyl-carbonyl, C 1-C 6Alkoxy carbonyl.
6. compound as claimed in claim 1 is characterized in that, described compound is selected from the compound of Formula Il I:
Figure FDA0000099751700000021
R 2Be selected from C 1-C 6Alkyl, the C that halogen replaces 1-C 6Alkyl, the optional phenyl that replaces, C 1-C 3Alkyl-carbonyl, the optional benzoyl that replaces, carboxyl, formamyl, C 1-C 6Alkoxy carbonyl, hydroxyl, C 1-C 6Alkoxyl group, optional aryl or the heteroaryl that replaces, amino, the optional amino that replaces;
R 3Be selected from C 1-C 6Alkyl, the optional phenyl that replaces, C 1-C 3Alkyl-carbonyl, C 1-C 3Alkoxy carbonyl, the optional benzoyl that replaces, carboxyl, amide group, the optional aniline formyl radical that replaces;
R 4Be selected from C 5Above alkyl or cycloalkyl, the optional phenyl that replaces, 5 or 6 Yuans fragrant heterocyclic radicals.
7. compound as claimed in claim 1 is characterized in that, described compound is selected from:
Figure FDA0000099751700000022
Figure FDA0000099751700000031
Figure FDA0000099751700000041
Figure FDA0000099751700000051
Figure FDA0000099751700000071
8. a pharmaceutical composition is characterized in that, described pharmaceutical composition contains each described compound or its pharmacy acceptable salt among the claim 1-7, and pharmaceutically acceptable carrier or vehicle.
9. the purposes of each described compound in the medicine that the disease of preparation treatment or the two clear vitamin B13 desaturases mediations of prevention is used among the claim 1-7.
10. each described compound is preparing the purposes that suppresses in pair medicine that clear vitamin B13 dehydrogenase activities are used among the claim 1-7.
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