CN104529972A - Protein kinase inhibitor namely coumarin as well as preparation method and medical application thereof - Google Patents

Protein kinase inhibitor namely coumarin as well as preparation method and medical application thereof Download PDF

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Publication number
CN104529972A
CN104529972A CN201510026529.5A CN201510026529A CN104529972A CN 104529972 A CN104529972 A CN 104529972A CN 201510026529 A CN201510026529 A CN 201510026529A CN 104529972 A CN104529972 A CN 104529972A
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Prior art keywords
trifluoromethyl
compound
hydroxyl
clocoumarol
protein kinase
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CN104529972B (en
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张娜
周玥
陈文娟
钟儒刚
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Beijing University of Technology
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Beijing University of Technology
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/06—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
    • C07D311/08—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring
    • C07D311/16—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted in position 7

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

The invention discloses a protein kinase inhibitor namely coumarin as well as a preparation method and a medical application thereof, and belongs to the technical field of medicinal chemistry. The structural formula of 4-trifluoromethyl-7-hydroxy-8-chlorocoumarin is shown in the specification, and by virtue of in vitro protein kinase activity and antitumor cell proliferation activity detections, the compound can be used for inhibiting the activity of protein kinase CK2, has a certain inhibiting effect on proliferation of lung cancer A549, cervical cancer Hela and mammary cancer MCF-7 cells, and can be used as a protein kinase inhibitor and an antitumor drug.

Description

Kinases inhibitor tonka bean camphor and preparation method thereof and medical applications
Technical field
The invention belongs to field of pharmaceutical chemistry technology, be specifically related to the target design of kinases inhibitor tonka bean camphor, and preparation method thereof and its application in anti-tumour cell proliferative.
Background technology
Protein kinase is one of protein family maximum in cell, altogether containing more than 500 protein kinase gene in human genome.It wants function to be transferred on the specific amino acid of substrate protein by the γ phosphate of ATP, and then regulates the activity of substrate protein, plays a crucial role in adjustment metabolism, genetic expression, Growth of Cells, division and differentiation etc.Closely related with tumorigenesis, become popular anticancer target spot, and existing more than ten plant kinases inhibitor and successfully go on the market as cancer therapy drug.Protein kinase C K2 is a kind of multi-functional serine/threonine proteinoid kinases, pass through phosphorylates various substrates, participate in the physiological processs such as the activity of oncogene and cancer suppressor gene in induced cell growth, inhibited apoptosis and conditioning signal path, there is short growth and the characteristic of anti-apoptotic and become important anticancer target.People source CK2 holoenzyme structure is the heterogeneity tetramer regulating subunit β to form by two catalytic subunits (α and/or α ') and two.Wherein CK2 α holds, is rich in the C of α-helixstructure end and the catalytic activity pocket between two structural domains to form by the N being rich in beta sheet.
Coumarin kind compound and derivative thereof are extremely extensive in occurring in nature distribution, have the biological activitys such as antibacterial, anti-inflammatory, antitumor, resist coagulation, have potential pharmaceutical use, be identified as important lead compound.Prior art shows, umbelliferone compounds only forms single polarity effect with active pocket, therefore show moderate inhibit activities to protein kinase C K2.For umbelliferone compound scaffold, appropriate the position of substitution is selected to carry out modified with functional group, to obtaining the compound with high CK2 inhibit activities and anti-tumour cell proliferative activity.
Summary of the invention
The present invention relates to 4-trifluoromethyl-7-hydroxyl-8-Clocoumarol and preparation method thereof, and the application in kinases inhibitor and anti-tumour cell proliferative field of medicaments.
ATP competitive inhibitor plays inhibit feature by being attached to catalytic activity pocket.Wherein the catalytic activity pocket of this target enzyme is primarily of hinge area (Glu114, Val116), positive polarity district (Lys68, Glu81 and water molecules 1) and hydrophobic region (Val66, F113, Met163 and Ile174) three sublocus compositions.For only forming single polarity effect umbelliferone compounds with CK2 catalytic activity pocket positive polarity district, the present inventor proposes to be ensure that compound has the important feature basis of high inhibit activities with binding pocket each sublocus formation effect (especially can form polarity effect with hinge area and positive polarity district) simultaneously.Therefore, the present inventor, based on umbelliferone skeleton structure, for increasing the polarity effect of compound and hinge region amino acid Glu114 and Val116, selecting appropriate the position of substitution to carry out suitable modified with functional group, proposing at R 4cF is introduced in position 3substituting group, to obtain the compound with high protein kinase c K2 inhibit activities and anti-tumour cell proliferative activity.
The invention provides-the 7-of 4-trifluoromethyl shown in structural formula 1 hydroxyl-8-Clocoumarol or its pharmacologically acceptable salt (4-trifluoromethyl-7-hydroxyl-8-Clocoumarol of the present invention, when its hydroxyl exists with negative oxygen ion form, can be combined with positively charged ion and form compound salt, as sodium salt, sylvite, ammonium salt etc.).
The structural formula of 4-trifluoromethyl-7-hydroxyl-8-Clocoumarol is as follows:
Coumarin compound preparation method shown in structural formula 1: by compound 2 (2-chloro-1,3-dihydroxy-benzene) be scattered in inorganic acid aqueous solution, add compound 3 (trifluoroacetic ethyl acetoacetate) subsequently, react under the condition that this reaction solution under agitation heats, after reacting completely, return to room temperature, separatory.Washing, dry, be separated, obtain obtained structural formula 1.
The mass concentration 60%-80% of preferred inorganic acid aqueous solution.The temperature to 60 of heating DEG C-90 DEG C; React 1 hour-3 hours.
Preferred compound 2: inorganic acid aqueous solution: compound 3 is (0.2-0.6) g:(1-3) mL:(0.20-0.40) mL.
Chemical equation is as follows:
Further, described mineral acid is any one in hydrochloric acid, sulfuric acid and perchloric acid, with saturated sodium bicarbonate and ethyl acetate separatory.Organic phase with after saturated common salt water washing, with anhydrous magnesium sulfate drying, column chromatography for separation (normal hexane: ethyl acetate=2:1).
Coumarin compound shown in structural formula 1 or its pharmacologically acceptable salt are as the application of kinases inhibitor.
Further, the coumarin compound shown in structural formula 1 or its pharmacologically acceptable salt as protein kinase C K2 inhibitor, its IC 50value is 0.4 μM, is better than not by coumarin kind compound that trifluoromethyl replaces.
Coumarin compound shown in structural formula 1 or its pharmacologically acceptable salt are as the application in antitumor drug.
Further, the coumarin compound shown in structural formula 1 has the activity of certain anti-tumour cell proliferative.Tumour cell comprises one or more in lung cell A549, cervical cancer cell Hela and breast cancer cell MCF-7.
Embodiment
Below in conjunction with embodiment the present invention will be further described book, but the present invention is not limited to following examples.
Embodiment 1
The preparation method of compound shown in structural formula 1
Compound 2 (the chloro-Resorcinol of 2-) (0.400g) is scattered in the perchloric acid of 1.5mL 70%, adds subsequently compound 3 (trifluoroacetic ethyl acetoacetate) (0.36mL).This reaction solution is under agitation heated to 80 DEG C, returns to room temperature after 2 hours, with saturated sodium bicarbonate and ethyl acetate separatory.Organic phase is with after saturated common salt water washing, and with anhydrous magnesium sulfate drying, column chromatography for separation (normal hexane: ethyl acetate=2:1), obtaining (0.39g) shown in obtained structural formula 1 is light yellow solid, and productive rate is 72%. 1HNMR(400MHz,DMSO-d 6):11.74(br s,1H),7.52(dd,1H,J＝1.9Hz,J＝9.0Hz),7.10(d,1H,J＝9.0Hz),6.86(s,1H).HRMS(ESI)m/z calculated for C 10H 4O 3F 3Cl[M+H] +:263.9801,found:263.9733。
Perchloric acid is replaced with other the mineral acid such as hydrochloric acid, sulfuric acid, same compound can be obtained equally.
Embodiment 2
Compound arrestin kinase c K2 determination of activity shown in structural formula 1.
Adopt the Non-radioactive methods detection compound of Kinase-Glo kinases luminescence detection kit (production of Promega company) to the inhibit activities of CK2.By kinase buffer liquid (0.1 μ g/L protein kinase C K2,1mM peptide substrate HRRRDDD-SDDD-NH 2, 2mM MnCl 2, 2mM DTT and 100 μMs of NaVO 3) be set to negative control, in 96 hole microtest plates, add compound solution shown in the diluted structural formula of the present invention 1 of kinase buffer liquid and positive inhibitor (compound 4) contrast solution respectively.Add 10 μMs of ATP and start reaction, hatch 30min in 30 DEG C.Every hole adds 25 μ LADP-Glo reagent termination reactions, adds 50 μ L kinase assay reagent and ADP is changed into ATP and hatches 1h, uses microwell plate luminometer measure and record the luminous value in every hole.With the logarithm of concentration for X-coordinate, respective concentration inhibiting rate is ordinate zou, and the activity of protein kinase CK 2 drawn with drug level change suppresses curve, tries to achieve IC 50value, the results are shown in Table 1.
Embodiment 3
Compound antitumor cell-proliferation activity shown in structural formula 1 measures.
CCK-8 reagent method is adopted to measure its restraining effect to lung cancer A549, cervical cancer cell lines Hela and breast cancer cell line MCF-7.The tumour cell being in cell log vegetative period is made into cell suspension, is inoculated in 96 orifice plates with 7500, every hole cell concentration.Hatch and cultivate 24h, arrange negative control 3 hole, every hole, other holes adds the testing compound (being respectively compound 1 and compound 4) of different concns, cultivates 48h.Every hole adds CCK-8 reagent, continues to cultivate 4h, stops cultivating.Add DMSO to every hole dissolving crystallized, in 490-570nm wavelength region, measure its absorbance value and curve plotting by microplate reader, indirect reaction cell survival quantity.
Table 1 coumarin compound arrestin kinase c K2 activity and anti-tumour cell proliferative activity (μM)
Can be found by the compound 1 and 4 in contrast table 1 and inhibit activities data thereof, R in the present invention 4for CF 3the R only forming single polarity effect with positive polarity district that obtains with prior art of compound 1 4for CH 3compound 4 compare, all have stronger arrestin kinase c K2 and suppressing lung cancer A 549, cervical cancer Hela cells and proliferation of MCF-7 cells are active.Its reasons in structure is mainly due to R 4cH 3be transformed to CF 3after, add the polarity effect with CK2 catalytic activity pocket hinge area, thus improve compound 1 inhibit activities.

Claims (10)

1.4-trifluoromethyl-7-hydroxyl-8-Clocoumarol or its pharmacologically acceptable salt, wherein the structural formula of 4-trifluoromethyl-7-hydroxyl-8-Clocoumarol is as follows:
The preparation method of 2.4-trifluoromethyl-7-hydroxyl-8-Clocoumarol, it is characterized in that: by chloro-for compound 2-1,3-dihydroxy-benzene 2 is scattered in inorganic acid aqueous solution, add compound trifluoroacetic ethyl acetoacetate 3 subsequently, react under the condition that this reaction solution under agitation heats, after reacting completely, return to room temperature, separatory.Washing, dry, be separated, obtain final product; Reaction formula is as follows:
3. according to the method for claim 2, it is characterized in that, the mass concentration 60%-80% of inorganic acid aqueous solution; The temperature to 60 of heating DEG C-90 DEG C; React 1 hour-3 hours.
4. according to the method for claim 2, it is characterized in that, mineral acid is any one in hydrochloric acid, sulfuric acid and perchloric acid.
5. according to the method for claim 2, it is characterized in that, compound 2: inorganic acid aqueous solution: compound 3 is (0.2-0.6) g:(1-3) mL:(0.20-0.40) mL.
6., according to the method for claim 2, it is characterized in that, with saturated sodium bicarbonate and ethyl acetate separatory, organic phase with after saturated common salt water washing, with anhydrous magnesium sulfate drying, normal hexane: the volume ratio=2:1 of ethyl acetate carries out column chromatography for separation.
7.4-trifluoromethyl-7-hydroxyl-8-Clocoumarol or its pharmacologically acceptable salt are as the application of kinases inhibitor.
8. according to the 4-trifluoromethyl-7-hydroxyl-8-Clocoumarol of claim 7 or its pharmacologically acceptable salt as the application of kinases inhibitor, it is characterized in that, protein kinase is CK2.
9.4-trifluoromethyl-7-hydroxyl-8-Clocoumarol or its pharmacologically acceptable salt are as the application in antitumor drug.
10. according to the 4-trifluoromethyl-7-hydroxyl-8-Clocoumarol of claim 9 or its pharmacologically acceptable salt as the application in antitumor drug, it is characterized in that, comprise one or more in lung cell A549, cervical cancer cell Hela and breast cancer cell MCF-7.
CN201510026529.5A 2014-10-10 2015-01-20 Kinases inhibitor cumarin and preparation method thereof and medical applications Expired - Fee Related CN104529972B (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104606186A (en) * 2014-11-19 2015-05-13 北京工业大学 Application of fluoro-7-hydroxycoumarin compound in protein kinase inhibitor or/and antitumor drug
CN112300106A (en) * 2020-10-30 2021-02-02 徐州医科大学 Novel protein kinase inhibitor, preparation method and application

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4737517A (en) * 1983-07-29 1988-04-12 Fidia, S.P.A. Coumarin derivatives, pharmaceutical compositions containing the same, and the use thereof in the treatment of cancer

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4737517A (en) * 1983-07-29 1988-04-12 Fidia, S.P.A. Coumarin derivatives, pharmaceutical compositions containing the same, and the use thereof in the treatment of cancer

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
ADRIANA CHILIN 等: "Coumarin as Attractive Casein Kinase 2 (CK2) Inhibitor Scaffold: An Integrate Approach To Elucidate the Putative Binding Motif and Explain Structure-Activity Relationships", 《J. MED. CHEM.》, vol. 51, no. 4, 6 February 2008 (2008-02-06), XP002656336, DOI: doi:10.1021/jm070909t *
ADRIANA CHILIN 等: "Coumarin as Attractive Casein Kinase 2 (CK2) Inhibitor Scaffold: An Integrate Approach To Elucidate the Putative Binding Motif and Explain Structure–Activity Relationships", 《J. MED. CHEM.》, vol. 51, no. 4, 6 February 2008 (2008-02-06), XP002656336, DOI: doi:10.1021/jm070909t *
NA ZHANG 等: "Docking and 3D-QSAR studies of 7-hydroxycoumarin derivatives as CK2 inhibitors", 《EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY》, vol. 45, no. 1, 13 October 2009 (2009-10-13) *
XIAO Y. HUANG 等: "Study on the Anticancer Activity of Coumarin Derivatives by Molecular Modeling", 《CHEM BIOL DRUG DES》, vol. 78, no. 4, 6 September 2011 (2011-09-06) *
谈云龙 等: "8(或6)-氯-7-羟基-4-甲基香豆素及其衍生物的合成", 《合成化学》, vol. 17, no. 3, 31 March 2009 (2009-03-31) *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104606186A (en) * 2014-11-19 2015-05-13 北京工业大学 Application of fluoro-7-hydroxycoumarin compound in protein kinase inhibitor or/and antitumor drug
CN112300106A (en) * 2020-10-30 2021-02-02 徐州医科大学 Novel protein kinase inhibitor, preparation method and application
CN112300106B (en) * 2020-10-30 2023-03-03 徐州医科大学 Protein kinase inhibitor, preparation method and application

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