CN102453016A - N-(2,6-dibenzyloxy benzoyl)-N'-substituted urea compound and preparation method and application thereof - Google Patents
N-(2,6-dibenzyloxy benzoyl)-N'-substituted urea compound and preparation method and application thereof Download PDFInfo
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- VMVKBYOKFOFDOD-UHFFFAOYSA-N N#Cc(c(OCc1ccccc1)ccc1)c1OCc1ccccc1 Chemical compound N#Cc(c(OCc1ccccc1)ccc1)c1OCc1ccccc1 VMVKBYOKFOFDOD-UHFFFAOYSA-N 0.000 description 1
- IZLPPMNAXBHFTI-UHFFFAOYSA-N O=C(c(c(OCc1ccccc1)ccc1)c1OCc1ccccc1)NC(NC1c2ccccc2CC1)=O Chemical compound O=C(c(c(OCc1ccccc1)ccc1)c1OCc1ccccc1)NC(NC1c2ccccc2CC1)=O IZLPPMNAXBHFTI-UHFFFAOYSA-N 0.000 description 1
- ZQICGJOWFRJEEY-UHFFFAOYSA-N O=C(c(c(OCc1ccccc1)ccc1)c1OCc1ccccc1)NC(Nc(cc1)ccc1F)=O Chemical compound O=C(c(c(OCc1ccccc1)ccc1)c1OCc1ccccc1)NC(Nc(cc1)ccc1F)=O ZQICGJOWFRJEEY-UHFFFAOYSA-N 0.000 description 1
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Abstract
The invention relates to a novel N-(2,6-dibenzyloxy benzoyl)-N'-substituted urea compound and a preparation method and an application thereof. Structural general formula of the compound is shown as a formula (I), wherein R1 is a C5-C25 cyclic alkyl, aryl or heterocyclic group, or a substituted C1-C6 straight-chain or branch-chain alkyl (shown in the specification). The current study shows that cyclophilin A (CypA) participates in multiple physiological and pathological processes of a human body. Thus, the compound provided by the invention can be applied to preparation of a cyclophilin A (CypA) excitant or preparation of drugs used for preventing or treating diseases mediated by CypA.
Description
Technical field
The present invention relates to pharmaceutical chemistry and chemicobiology field; Be specifically related to the N-(2 of one type of non-peptide excitomotor of the small molecules as cyclophilin A; 6-benzyloxy benzoyl-)-N '-substitute urea compound and preparation method thereof and biological applications; This compounds can be used as the small molecules probe, is used for the research of cyclophilin A Mediated Signal Transduction path.
Background technology
Cyclophilin (Cyclophilin; CyP) discovery and name originate from it, and (CsA is widely used clinically a kind of immunosuppressive drug (.Science such as Handschumacher RE for Cyclosporin A, height affinity CsA) to ciclosporin; 1984,226:544-547).CyP be a conservative protein families (Gothel SF, Marahiel MA.Cell Mo.Life Sci, 1999,55:423-436), wait prokaryotic organism this proteinoid all to be arranged to Mammals from low.The common trait of this histone be have peptide cis-trans propyl isomerism enzyme (Peptidyl-prolyl cis-trans isomerase, PPIase) active (Takahashi N, Hayano T, Suzuki M.Nature, 1989,337:473-475).According to characteristics such as relative molecular weight and Subcellular Localization, there are 16 kinds of members such as CypA, CypB, CypC and CypD in known person CyP family at present, and people's cyclophilin A (hCypA) is most important; Also be to study member (Braaren D, Luban J.EMBO J, 2001 the most widely; 20:1300-1309), it is made up of 165 amino acid, relative molecular weight 18; 000, account for 0.1%～0.4% of cellular proteins total amount, mainly be positioned at kytoplasm.Except conjugated protein in the body as CsA, in vivo and in vitro, hCypA can both conjugated protein and catalysis quicken their folding, assembling and transhipment, play a part Chaperones Molecular (.Science such as Freskgard PO, 1992,258:466).
CypA comprises stereotyped writing antiparallel β lamella; These β lamellas are formed a barrel-like structure; Upper and lower opening at bucket respectively has the closed at both ends of one section α spiral with bucket, and the part binding pocket of CypA then is positioned at side (.Nature such as Kallen J, 1991 of this barrel-like structure; 353:276-279), mainly form: inferior pocket A and inferior pocket B by two big inferior pockets.Inferior pocket A mainly is made up of residues such as Thr73, Gly74, Asn102, Ala103, Thr107, Gly109, Ser110 and Gln111, and their major parts are the wetting ability residue; Inferior pocket B is a dark hydrophobicity groove, mainly is made up of residues such as Phe60, Met61, Phe113 and Leu122.
CypA and multiple disease are closely related; 1993, people such as Luban found that CypA and CypB can combine with the Gag protein-specific ground of I type HIV (HIV-1), and CsA can suppress the interaction (.Cell such as Luban J between the Gag-CypA effectively; 1993,73:1067-1078); 1994, Franke (Franke EK, Yuan HE.Nature; 1994,372:359-362) with people such as Thali (.Nature such as Thali M, 1994; 372:363-365) report that on Nature, publishes thesis respectively finds that CypA is incorporated in the HIV-1 virion specifically, and blocking-up CypA mixes, and suppresses duplicating of HIV-1; This explanation CypA and the proteic combination of Gag be function relevant (Luban J.Cell, 1996,87:1157-1159).The Gag albumen of HIV-1 is a polyprotein, it in virus assembling, the ripe and process of sprouting, play decisive role (.Cell such as Gamble TR, 1996,87:1285-1294).Capsid protein (Capsid, CA in CypA and the Gag albumen; 231residues) zone combines, and through this effect, CypA is wrapped up in the HIV-1 virion specifically.But the disappearance of CypA do not influence virus assembling, sprout and ripe, it is essential at early stage (the viral rt begin before) of HIV-1 life cycle.Therefore, CypA possibly play a role in shelling (Uncoating) process after virus gets into host cell, helps nucleoid to separate assembling (Viral CA core disassembly).
The Jiang Hua of Shanghai Pharmaceutical Inst., Chinese Academy of Sciences very waits people (.Biochem Biophys Res Commun such as Luo C; 2004; 321:557-565) find: CA albumen though SARS-CoV does not encode through the means of information biology; But the proteic Val235 of SARS-CoVN～Pro369 segment and the proteic N end structure of HIV-1CA territory (residue 1～151) have higher sequence similarity (36.7%), therefore infer that SARS-CoV N albumen possibly also interact with CypA.Afterwards, protein-protein combines activity experiment to confirm this hypothesis: SARS-CoV N albumen and CypA have very high affinity, and the KD value is about 6～160nM; Ensuing rite-directed mutagenesis experiment has confirmed that tentatively the proteic Trp302 of SARS-CoVN～Pro310 segment is for regional with the CypA bonded.Above presentation of results CypA probably participates in the vital movement process of SARS-CoV.
The mixture that hCypA and part form has provide protection to multiple neurone, for aixs cylinder, myelin after the damage all have significant short regeneration (Yoshimoto T, Siesjo BK.Brain Res, 1999,839:283-291).Confirmation CypA such as Andreeva are a kind of growth factors of oxidative stress secretion inducing; Mediation active oxygen medium pair cell external signal is regulated the activation and the vascular smooth muscle cell proliferation of kinases (ERK1/2); Show CypA in the vascular disease pathogenesis, play a significant role (Andreeva L, Heads R, Green CJ.Int J Exp Path; 1999,80:305-315).
Except having above-mentioned physiological effect, hCypA is the most deep of research about the effect of immunosuppression aspect.The immunosuppressive action mechanism of generally acknowledging at present is: hCypA combines with intracytoplasmic part, the mixture of formation then with calcineurin (Calcineurin, CaN) phosphorylation activity of combination and inhibition CaN (.Cell such as Liu J, 1991,66:807-815).CaN has important effect in lymphocytic reactivation process; It is through dephosphorylation activating T cell nf (Nuclearfactor of activated T cell; NF-AT), make it activation and in the entering nuclear that is shifted, participate in the transcription of IL-2 and some other factor then.The phosphorylation activity that has suppressed CaN has just suppressed to have the release of the cytokine of immunization, thus the performance immunosuppressive action.
In view of CypA plays a role in a plurality of physiological pathway, therefore the suppressor factor research about it is a focus always.Divide by structure, the CypA suppressor factor is divided into three major types: natural product and analogue thereof, peptide class and plan peptide compounds and organic synthesis micromolecular compound.In order to overcome the toxic side effect of natural product, and the easy metabolic of peptides, seeking high, the active strong organic synthesis small molecules CypA suppressor factor of selectivity becomes inevitable.At present existing a plurality of study group have reported the small molecules CypA suppressor factor of several kinds of structure types.
Though the discovery of CypA suppressor factor and biological applications thereof have been obtained bigger progress, CypA agonist and potential biological applications thereof are blank always, do not see any bibliographical information so far.In view of CypA extensive and important effect in multiple Human Physiology and pathologic process; Through exploitation CypA agonist; Unite existing CypA suppressor factor; Can carry out positive down regulation research, further deeply effectively explore the research of CypA Mediated Signal Transduction path the CypA catalytic activity.
In the process of system design CypA suppressor factor; The contriver has found a N-(2; 6-benzyloxy benzoyl-)-N '-replacement carbamide compound; Utilization rational drug method of design is carried out structural modification to this compound and is obtained a compounds, and has tested the activity of this compounds exciting CypA in the horizontal experiment test of vitro enzyme.The result proves that the application's part of compounds has stronger CypA agonist activity.
Summary of the invention
An object of the present invention is: a kind of novel N-(2,6-benzyloxy benzoyl-)-N '-substitute urea compound is provided, and its general structure is suc as formula shown in the I:
Among the formula I: R
1Be C
5～C
25Cyclic hydrocarbon radical, aromatic ring yl or heterocyclic radical, or substituted C
1～C
6The straight or branched alkyl;
Wherein: said substituted C
1～C
6The substituting group of alkyl is: replace or unsubstituted phenyl or naphthyl, and the substituting group of said substituted phenyl or naphthyl is selected from: halogen (F, Cl, Br or I), C
1-C
6The straight or branched alkyl, hydroxyl, C
1-C
4Alkoxyl group, C
1-C
4Unsaturated-oxyl, carboxyl, the C of carboxyl substituted
1-C
6Alkoxyl group, cyanic acid, nitro, amino, trifluoromethyl, trifluoromethoxy, sulfydryl or C
1-C
4A kind of or (contain two kinds) more than two kinds in the acyl group, substituent number is 1～4;
The heteroatoms of said heterocyclic radical is selected from: a kind of or (contain two kinds) more than two kinds among N, O or the S, the heteroatoms number is 1～3.
In optimized technical scheme of the present invention,
R
1Be selected from: a kind of in the group shown in formula II, III, IV, V, VI or the VII:
Wherein, X is N, O or S; M is 0～2 integer;
R
2And R
3Be selected from independently of one another: hydrogen (H), C
1-C
6The alkyl of straight or branched, C
1-C
4Alkoxyl group, halogen (F, Cl, Br or I), cyanic acid, nitro, amino, hydroxyl, trifluoromethyl, carboxyl, sulfydryl or C
1-C
4A kind of in the acyl group;
Ar is for replacing or unsubstituted phenyl or naphthyl, and the substituting group of said substituted phenyl or naphthyl is selected from: halogen (F, Cl, Br or I), C
1-C
6The straight or branched alkyl, hydroxyl, C
1-C
4Alkoxyl group, C
1-C
4Unsaturated-oxyl, carboxyl, the C of carboxyl substituted
1-C
6Alkoxyl group, cyanic acid, nitro, amino, trifluoromethyl, trifluoromethoxy, sulfydryl or C
1-C
4A kind of or (contain two kinds) more than two kinds in the acyl group, substituent number is 1～4.
In another optimized technical scheme of the present invention, R
1Be group shown in formula III or the VI; M is 0～2 integer;
Ar is for replacing or unsubstituted phenyl or naphthyl, and the substituting group of said substituted phenyl or naphthyl is selected from: halogen (F, Cl, Br or I), cyanic acid, nitro, trifluoromethyl, trifluoromethoxy, sulfydryl or C
1-C
4A kind of or (contain two kinds) more than two kinds in the acyl group, substituent number is 1～4;
In a preferred technical scheme, R
1Be group shown in formula III or the VI; M is 0;
Ar is for replacing or unsubstituted phenyl or naphthyl, and the substituting group of said substituted phenyl or naphthyl is selected from: halogen (F, Cl, Br or I), cyanic acid, nitro, trifluoromethyl, trifluoromethoxy, sulfydryl or C
1-C
4A kind of or (contain two kinds) more than two kinds in the acyl group, substituent number is 1～4;
Best R
1Be fluorenyl, naphthyl, phenyl or 3-chloro-4-fluorophenyl.
In another optimized technical scheme of the present invention, described N-(2,6-benzyloxy benzoyl-)-N '-substitute urea compound specifically:
N-(2,6-benzyloxy benzoyl-)-N '-(1-xanthenyl)-urea, N-(2,6-benzyloxy benzoyl-)-N '-cyclohexyl-urea, N-(2; 6-benzyloxy benzoyl-)-N '-(2; 3-dihydro-indenes-1-yl)-urea, N-(2,6-benzyloxy benzoyl-)-N '-phenmethyl-urea, N-(2,6-benzyloxy benzoyl-)-N '-styroyl-urea, N-(2; 6-benzyloxy benzoyl-)-N '-(4-fluorophenyl)-urea, N-(2; 6-benzyloxy benzoyl-)-N '-diphenyl-methyl-urea, N-(2,6-benzyloxy benzoyl-)-N '-fluorenyl-urea, N-(2,6-benzyloxy benzoyl-)-N '-naphthyl-urea, N-(2; 6-benzyloxy benzoyl-)-N '-phenyl-urea or N-(2,6-benzyloxy benzoyl-)-N '-(3-chloro-4-fluorophenyl)-urea.
Another object of the present invention is: the method for the above-mentioned N-of a kind of preparation (2,6-benzyloxy benzoyl-)-N '-substitute urea compound is provided, and said method comprises step (i), (ii) and (iii):
(i) by 2,6-difluorobenzonilyile preparation 2, the step of 6-benzyloxy BM;
(ii) by R
1-NH
2Preparation R
1The step of-NCO; With
(iii) under reflux state, by 2,6-benzyloxy BM and R
1-NCO carries out addition reaction and gets target compound;
Wherein: R
1Definition said identical with preamble, step (i) with (ii) do not have strict order requirement, its order is interchangeable, even carries out simultaneously.
Said preparing method's synthetic route is following:
Wherein, R
1Definition said identical with preamble, Bn representes benzyl.
A further object of the present invention is: disclose a kind of N-according to the invention (2; 6-benzyloxy benzoyl-)-purposes of N '-substitute urea compound (compound shown in the formula I); Be the application of compound shown in the formula I in preparation CypA agonist, or the application of compound shown in the formula I in the medicine of the disease of preparation prevention or treatment CypA mediation.
Description of drawings
Fig. 1 is that compound I-10 pair CypA enzyme is lived when exciting-the effect graphic representation.A is for adding Chymotrypsin (substrate of specificity hydrolysis trans-configuration) and reaction substrate (Suc-Ala-Ala-cis-Pro-Phe-pNA (AAPF)) among the figure; B is for adding CypA (promoting that the cis-configuration conversion in the reaction solution is the trans-configuration), Chymotrypsin and reaction substrate; C is for adding compound I-10, CypA, Chymotrypsin and substrate.
Embodiment
A kind of method for preparing compound shown in the formula I provided by the present invention, it comprises the steps:
1) with benzylalcohol sodium and 2, the 6-difluorobenzonilyile is dissolved in the methyl-sulphoxide, stirs 5 hours～10 hours after being warming up to 110 ℃～130 ℃, is chilled to room temperature; In reaction solution, add frozen water, stirred 30 minutes, separate out a large amount of solids; Suction filtration, washing, drying; Get white solid, recrystallization gets the white plates crystal 2,6-dibenzyloxy benzene nitrile (brief note: compound VIII);
2) will be dissolved in the benzylalcohol by the compound VIII that step 1) obtains, and add Pottasium Hydroxide and less water, in 120 ℃～140 ℃ stirrings 10 hours～15 hours, underpressure distillation removed and desolvates; In resistates, add frozen water, stirred 30 minutes, separate out a large amount of solids, suction filtration; Washing, drying gets white solid; Use silica gel column chromatography separating purification, obtain white powder solid 2,6-benzyloxy BM (brief note: compound I X);
3) TRIPHOSGENE 99.5 is dissolved in 1, in the 2-ethylene dichloride, ice bath is cooled to 0 ℃～5 ℃, will be dissolved in 1, the R of 2-ethylene dichloride
1-NH
2(R
1Substitutional amine-group compound) slowly be added dropwise in the reaction solution that contains TRIPHOSGENE 99.5, finish, continue the ice bath cooling and stirred 30 minutes down, after be warming up to room temperature; And, be heated to backflow then room temperature state maintenance at least 1 hour, when reaction solution is the water white transparency shape, triethylamine is added in the reaction solution; Continue to reflux 30 minutes, underpressure distillation removes and desolvates, and adds toluene; Suction filtration, the filtrating evaporate to dryness obtains faint yellow solid, is the substituted isocyanic ester of R1 (brief note: compounds X);
4) will be by step 2) and step 3) the compound I X and the X that obtain dissolve in the toluene; Be heated to backflow, and kept 10 hours～15 hours, remove reaction solvent under reduced pressure and obtain faint yellow solid at reflux state; Use silica gel column chromatography separating purification, obtain target compound (compound shown in the formula I).
Wherein said room temperature is: 20 ℃～25 ℃, down together, R
1Definition said identical with preamble.
Below through embodiment the present invention is done further elaboration, these embodiment only are used for better understanding the present invention, the protection domain that it does not limit the present invention in any way.
All parameters among the embodiment unless otherwise indicated, all are to be unit with quality (gram).
Embodiment 1
2, the preparation of 6-dibenzyloxy benzene nitrile
0.5 gram benzylalcohol sodium is dissolved in 10 milliliters of methyl-sulphoxides, stirs adding 2 down, the 6-difluorobenzonilyile.Be rapidly heated to 115 ℃ of continued stirrings 8 hours.Be chilled to room temperature, in 100 milliliters of frozen water impouring reaction solutions, stirred 30 minutes, separate out a large amount of solids, suction filtration, washing, drying gets white solid.Get white plates crystalline compounds VIII with ethyl alcohol recrystallization.
1H-NMR(400Hz，DMSO-d
6)δ：5.19(4H，s，-CH
2)，6.60(2H，d，Ar-H)，7.30-7.35(3H，m，Ar-H)，7.38(4H，t)，7.45(4H，d)。
2, the preparation of 6-benzyloxy BM
With 0.4 gram 2,6-dibenzyl benzene nitrile (VIII) is dissolved in 7 milliliters of benzylalcohols, and adds 0.6 gram KOH and 0.1 ml water, and 130 ℃ were stirred 12 hours down.Underpressure distillation removes and desolvates, and in 100 milliliters of frozen water impouring reaction solutions, stirs 30 minutes, separates out a large amount of solids, suction filtration, and washing, drying gets white solid.Silica gel column chromatography separates (petrol ether/ethyl acetate=12/1 (v/v)), obtains white powder solid chemical compound IX.
1H-NMR(400Hz，DMSO-d
6)δ：5.15(4H，s，-CH
2)，6.60(2H，d，Ar-H)，7.18-7.48(12H，m，Ar-H)；EI-MS?m/z?333.2(M
+)，91.1(100％)。
Embodiment 3
N-(2,6-benzyloxy benzoyl)-N '-(1-xanthenyl)-urea (compound (I-1))
0.5 gram TRIPHOSGENE 99.5 is dissolved in 3 milliliter 1, and in the 2-ethylene dichloride, ice bath is stirred to dissolving.0.25 gram xanthene-1-amine is dissolved in 10 milliliter 1, in the 2-ethylene dichloride, and slowly is added dropwise in the reaction solution that contains TRIPHOSGENE 99.5 the reaction solution muddiness that is white in color.Continue ice bath after 0.5 hour, be warming up to 25 ℃ of reactions 1 hour, when reflux to reaction solution is the water white transparency shape then, continuing to reflux 0.5 hour in 1 milliliter of triethylamine adding reaction solution.Underpressure distillation removes and desolvates, and adds 10 milliliters of toluene, suction filtration, and the filtrating evaporate to dryness obtains faint yellow solid, is xanthene-1-based isocyanate (X-1);
With above-mentioned xanthene-1-based isocyanate (X-1) and 0.25 gram 2,6-benzyloxy BM (IX) is dissolved in 10 milliliters of toluene, back flow reaction 10 hours.Remove solvent under reduced pressure, add elutriation and go out solid, it is used silica gel column chromatography separating purification, eluent is ethyl acetate/petroleum ether=1/4 (v/v), obtains white solid compound (I-1).Yield 44%.
1H-NMR(400Hz，DMSO-d
6)δ：5.13(4H，s，-CH)，6.39(1H，d，Ar-H)，6.56(2H，t，Ar-H)，6.81(2H，d，Ar-H)，7.03(2H，d，Ar-H)，7.19(2H，t，Ar-H)，7.29(3H，t，Ar-H)，7.39-7.50(10H，m，Ar-H)，9.13(1H，s，-NH)，10.15(1H，s，-NH)；EI-MS?m/z?556(M
+)。
N-(2,6-benzyloxy benzoyl)-N '-naphthyl-urea (compound (I-2))
Xanthene among the embodiment 3-1-amine is replaced to naphthalidine, and all the other desired raw materials, reagent and preparation method get pale solid compound (I-2) with embodiment 1～3.Yield 50%;
1H-NMR(400Hz，DMSO-d
6)δ：5.11(4H，s，-CH)，6.46(1H，d，-CH)，6.60(2H，t，Ar-H)，7.26(1H，s，Ar-H)，7.47-7.64(10H，m，Ar-H)，7.66-7.72(3H，m，Ar-H)，7.93-8.08(3H，m，Ar-H)，9.21(1H，s，Ar-H)，10.15(1H，s，-NH)；EI-MS?m/z?502(M
+)。
Embodiment 5
N-(2,6-benzyloxy benzoyl)-N '-phenmethyl-urea (compound (I-3))
Xanthene among the embodiment 3-1-amine is replaced to benzylamine, and all the other desired raw materials, reagent and preparation method get pale solid compound (I-3) with embodiment 1～3.Yield 52%.
1H-NMR(400Hz，DMSO-d
6)δ：4.42(2H，d，-CH2)，5.12(4H，s，-CH)，6.42(2H，d，Ar-H)，7.18-7.33(6H，m，Ar-H)，7.39-7.50(10H，m，Ar-H)，9.34(1H，s，-NH)，10.54(1H，s，-NH)；EI-MSm/z?466(M
+)。
N-(2,6-benzyloxy benzoyl)-N '-styroyl-urea (compound (I-4))
Xanthene among the embodiment 3-1-amine is replaced to phenylethylamine, and all the other desired raw materials, reagent and preparation method get khaki color solid chemical compound (I-4) with embodiment 1～3.Yield 67%.
1H-NMR(400Hz，DMSO-d
6)δ：2.80(2H，t，-CH2)，3.44(2H，q，-CH2)，5.12(4H，s，-CH)，6.41(2H，d，Ar-H)，7.21-7.30(6H，m，Ar-H)，7.39-7.50(10H，m，Ar-H)，8.34(1H，s，-NH)，10.54(1H，s，-NH)；EI-MS?m/z?480(M
+)。
Embodiment 7
N-(2,6-benzyloxy benzoyl)-N '-cyclohexyl-urea (compound (I-5))
Xanthene among the embodiment 3-1-amine is replaced to hexahydroaniline, and all the other desired raw materials, reagent and preparation method get pale solid compound (I-5) with embodiment 1～3.Yield 41%.
1H-NMR(400Hz，DMSO-d
6)δ：1.52-1.68(10H，m，-CH2)，3.60(1H，d，-CH)，5.12(4H，s，-CH)，6.42(2H，d，Ar-H)，7.22(1H，s，Ar-H)，7.39-7.50(10H，m，Ar-H)，8.24(1H，s，-NH)，10.52(1H，s，-NH)；EI-MS?m/z?458(M
+)。
N-(2,6-benzyloxy benzoyl)-N '-phenyl-urea (compound (I-6))
Xanthene among the embodiment 3-1-amine is replaced to aniline, and all the other desired raw materials, reagent and preparation method get white solid compound (I-6) with embodiment 1～3.Yield 40%.
1H-NMR(400Hz，DMSO-d
6)δ：5.15(4H，s，-CH)，7.02(2H，d，Ar-H)，7.10(2H，d，Ar-H)，7.30(1H，t，Ar-H)，7.34-7.53(10H，m，Ar-H)，7.62-7.78(3H，m，Ar-H)，8.69(1H，s，-NH)，11.23(1H，s，-NH)；EI-MS?m/z?452(M
+)。
Embodiment 9
N-(2,6-benzyloxy benzoyl)-N '-(4-fluorophenyl)-urea (compound (I-7))
Xanthene among the embodiment 3-1-amine is replaced to the 4-fluoroaniline, and all the other desired raw materials, reagent and preparation method get pale solid compound (I-7), yield 49% with embodiment 1～3.
1H-NMR(400Hz，DMSO-d
6)δ：5.13(4H，s，-CH)，6.74(2H，d，Ar-H)，7.10-7.21(3H，m，Ar-H)，7.35(2H，d，Ar-H)，7.44-7.63(10H，m，Ar-H)，8.80(1H，s，-NH)，11.02(1H，s，-NH)；EI-MS?m/z470(M
+)。
N-(2,6-benzyloxy benzoyl)-N '-(2,3-dihydro-indenes-1-yl)-urea (compound (I-8))
Xanthene among the embodiment 3-1-amine is replaced to 2,3-dihydro-indenes-1-amine, all the other desired raw materials, reagent and preparation method get light brown solid chemical compound (I-8) with embodiment 1～3, yield 82%,
1H-NMR(400Hz，DMSO-d
6)δ：1.65(2H，m，-CH2)，2.38(2H，m，-CH2)，5.03(1H，dd，-CH-)，5.13(4H，s，-CH)，6.78(2H，d，Ar-H)，7.17-7.30(5H，m，Ar-H)，7.37-7.55(10H，m，Ar-H)，8.54(1H，s，-NH)，11.65(1H，s，-NH)；EI-MS?m/z?492(M
+)。
Embodiment 11
N-(2,6-benzyloxy benzoyl)-N '-(diphenyl-methyl)-urea (compound (I-9))
Xanthene among the embodiment 3-1-amine is replaced to benzhydrylamine, and all the other desired raw materials, reagent and preparation method get khaki color solid chemical compound (I-9), yield 70% with embodiment 1～3.
1H-NMR(400Hz，DMSO-d
6)δ：5.14(4H，s，-CH)，6.08(1H，d，-CH-)，6.64(2H，d，Ar-H)，7.10-7.15(3H，m，Ar-H)，7.32-7.37(8H，m，Ar-H)，7.44-7.63(10H，m，Ar-H)，9.19(1H，s，-NH)，10.13(1H，s，-NH)；EI-MS?m/z?542(M
+)。
N-(2,6-benzyloxy benzoyl)-N '-fluorenyl-urea (compound (I-10))
Xanthene among the embodiment 3-1-amine is replaced to the 1-fluorenamine, and all the other desired raw materials, reagent and preparation method get khaki color solid chemical compound (I-10), yield 60% with embodiment 1～3.
1H-NMR(400Hz，DMSO-d
6)δ：5.12(4H，s，-CH)，6.43(1H，d，-CH-)，6.64(2H，d，Ar-H)，7.14-7.19(5H，m，Ar-H)，7.32-7.37(4H，m，Ar-H)，7.44-7.63(10H，m，Ar-H)，9.45(1H，s，-NH)，11.03(1H，s，-NH)；EI-MS?m/z?540(M
+)。
Embodiment 13
N-(2,6-benzyloxy benzoyl)-N '-(3-chloro-4-fluorophenyl)-urea (compound (I-11))
Xanthene among the embodiment 3-1-amine is replaced to 3-chloro-4-fluoroaniline, and all the other desired raw materials, reagent and preparation method get pale solid compound (I-11), yield 49% with embodiment 1～3.
1H-NMR(400Hz，DMSO-d
6)δ：5.20(4H，s，-CH)，6.68(2H，d，Ar-H)，7.10-7.21(2H，m，Ar-H)，7.35(2H，d，Ar-H)，7.40-7.59(10H，m，Ar-H)，9.81(1H，s，-NH)，11.12(1H，s，-NH)；EI-MSm/z?504(M
+)。
The compounds of this invention is to the percentile mensuration of CypA peptidyl cis-trans propyl isomerism enzyme agonist activity
(1) proteic expression of CypA and purifying
The plasmid PET15b-CypA that builds changed over to obtain expressing engineering bacteria in the e. coli bl21; Engineering bacteria is incubated at overnight cultures (peptone 10g/L in the 10mL LB substratum that contains 100 μ g/mL penbritins; Yeast extract 5 g/L, sodium-chlor 10g/L).Go into 1L by switching in 1: 100 then and contain in the fresh LB substratum of penbritin 37 ℃ of 220 rev/mins of cultivations.Work as OD
600Reach at about 0.8 o'clock, add IPTG (isopropyl ss-D thiogalactoside, 1M solution), reduce the temperature to 25 ℃ of cultivations simultaneously and carried out induction expression of protein in 5 hours to final concentration 0.5mM.4000 rev/mins of centrifugal 30 minutes collection thalline are put in-80 ℃ of Ultralow Temperature Freezers preservations and spend the night after collection is good.(the 5mM imidazoles pH8.0) hangs for 20mM Tris-Cl, 0.5M NaCl, with ultrasonic disruption on the suspension-s ice bath (300W worked 30 minutes, one time 5 seconds, midfeather 10 seconds) with the binding buffer liquid of 20mL with thalline.The cell homogenates that obtains after the fragmentation 4 ℃ with 10000 rev/mins centrifugal 30 minutes, keep supernatant.This protein crude extract injects and uses the good Ni-NTA resin of binding buffer liquid balance in advance afterwards, fully combines 1 hour at 4 ℃, uses the binding buffer liquid of 10 times of column volumes and cleaning buffer solution (the 20mM Tris-Cl of 10 times of column volumes more respectively; 0.5M NaCl, 60mM imidazoles, pH8.0) flushing resin; Remove the foreign protein that non-specific adsorption is arranged with resin, use elution buffer (20mM Tris-Cl, 0.5M NaCl at last; The 100mM imidazoles, pH 8.0) 10mL wash-out resin, the hCypA of collection wash-out.Detect proteic molecular weight and purity with SDS-PAGE.The albumen that dialysis is good can carry out enzyme activity determination.
(2) The compounds of this invention is to the mensuration of CypA agonist activity
(Biochemistry 1991,30 (25): 6127-6134), measure peptide prolyl cis-trans isomerase (PPIase) activity of target compound in establishing criteria Chymotrypsin coupling test.Whole test is carried out under 4 ℃ of constant temperature, and zymolyte is Suc-Ala-Ala-cis-Pro-Phe-pNA (AAPF), is dissolved into the mother liquor of 3.8mM with the THF that contains 400mM LiCl.Chymotrypsin is dissolved in the HCl solution of 1mM, is made into the mother liquor of 3mg/mL.At first to 519 μ L reaction buffer (50mM HEPES; 100mM NaCl) the middle testing compound solution (final concentration is 10 μ M) that adds 30 μ L hCyPA albumen mother liquors (final concentration 10 μ g/ml) and be dissolved in DMSO; In precooling on ice; They are added in the quartz curette, add 15 μ L Chymotrypsins again, add 12 μ L substrate initial actions at last.Whenever detected the variation of optical absorption intensity and collect data through the U-2010 spectrophotometer at 4 ℃ of following 380nM places at a distance from 0.1 second, after 1 minute, reaction finishes.Each reaction repetition 3 times.Utilize the slope (K) of the preceding 6 seconds linear growth of reaction to represent that the vigor of CypA enzyme is big or small.Draw the exciting rate of testing compound percentage under 10 μ M with formula,
Calculation formula is:
(experimental group K value-positive controls K value)/(positive controls K value-negative experimental group K value) * 100%
Wherein positive controls K value is not for adding testing compound institute measured value, and negative control group K value is not for adding testing compound and CypA institute measured value.
(3) compound agonist activity test result
Table 1N-(2,6-benzyloxy benzoyl-)-N '-substitute urea compound is to the exciting rate data of CypA
Can find out by table 1; N-(2 with general structure (I) of the present invention; 6-benzyloxy benzoyl-)-N '-substituted urea class part of compounds has tangible CypA agonist activity; Especially I-10 and the I-11 exciting percentage under 10 μ M concentration explains that near 50% compound of the present invention is the CypA agonist.
Compound I-10 pair CypA enzyme lives when exciting-and the effect graphic representation sees Fig. 1.By the time-the effect graphic representation can find out; This agonist can obviously improve the photon absorbing intensity (promptly showing the agonist activity to the CypA enzyme) at 380nM place, and the slope (K) of the linear growth in the reaction times of monitoring is the positive controls K value when not adding agonist obviously.
The possibility of utilizing on the industry
Advantage such as the preparation method of N-of the present invention (2,6-benzyloxy benzoyl-)-N '-substitute urea compound has that reaction conditions gentleness, abundant raw material are easy to get, operation and aftertreatment are simple.
N-of the present invention (2,6-benzyloxy benzoyl-)-N '-substitute urea compound has shown tangible exciting effect, has confirmed its Biological mechanism of action in CypA enzymatic activity testing experiment.Therefore, compound of the present invention can be used as the small molecules probe, is used for the research of CypA Mediated Signal Transduction path.
Claims (9)
1. a N-(2,6-benzyloxy benzoyl-)-N '-substitute urea compound, its general structure is suc as formula shown in the I:
Among the formula I: R
1Be C
5～C
25Cyclic hydrocarbon radical, aromatic ring yl or heterocyclic radical, or substituted C
1～C
6The straight or branched alkyl;
Wherein: said substituted C
1～C
6The substituting group of alkyl is: replace or unsubstituted phenyl or naphthyl, and the substituting group of said substituted phenyl or naphthyl is selected from: halogen, C
1-C
6The straight or branched alkyl, hydroxyl, C
1-C
4Alkoxyl group, C
1-C
4Unsaturated-oxyl, carboxyl, the C of carboxyl substituted
1-C
6Alkoxyl group, cyanic acid, nitro, amino, trifluoromethyl, trifluoromethoxy, sulfydryl or C
1-C
4A kind of or more than two kinds, substituent number is 1～4 in the acyl group;
The heteroatoms of said heterocyclic radical is selected from: a kind of or more than two kinds, the heteroatoms number is 1～3 among N, O or the S.
2. N-as claimed in claim 1 (2,6-benzyloxy benzoyl-)-N '-substitute urea compound is characterized in that, wherein R
1Be selected from: a kind of in the group shown in formula II, III, IV, V, VI or the VII:
Wherein, X is N, O or S; M is 0～2 integer;
R
2And R
3Be selected from independently of one another: hydrogen, C
1-C
6The alkyl of straight or branched, C
1-C
4Alkoxyl group, halogen, cyanic acid, nitro, amino, hydroxyl, trifluoromethyl, carboxyl, sulfydryl or C
1-C
4A kind of in the acyl group;
Ar is for replacing or unsubstituted phenyl or naphthyl, and the substituting group of said substituted phenyl or naphthyl is selected from: halogen, C
1-C
6The straight or branched alkyl, hydroxyl, C
1-C
4Alkoxyl group, C
1-C
4Unsaturated-oxyl, carboxyl, the C of carboxyl substituted
1-C
6Alkoxyl group, cyanic acid, nitro, amino, trifluoromethyl, trifluoromethoxy, sulfydryl or C
1-C
4A kind of or more than two kinds, substituent number is 1～4 in the acyl group.
3. N-as claimed in claim 2 (2,6-benzyloxy benzoyl-)-N '-substitute urea compound is characterized in that, wherein R
1Be group shown in formula III or the VI; M is 0～2 integer;
Ar is for replacing or unsubstituted phenyl or naphthyl, and the substituting group of said substituted phenyl or naphthyl is selected from: halogen, cyanic acid, nitro, trifluoromethyl, trifluoromethoxy, sulfydryl or C
1-C
4A kind of or more than two kinds, substituent number is 1～4 in the acyl group.
4. N-as claimed in claim 3 (2,6-benzyloxy benzoyl-)-N '-substitute urea compound is characterized in that, wherein R
1Be group shown in formula III or the VI; M is 0;
Ar is for replacing or unsubstituted phenyl or naphthyl, and the substituting group of said substituted phenyl or naphthyl is selected from: halogen, cyanic acid, nitro, trifluoromethyl, trifluoromethoxy, sulfydryl or C
1-C
4A kind of or more than two kinds, substituent number is 1～4 in the acyl group.
5. N-as claimed in claim 4 (2,6-benzyloxy benzoyl-)-N '-substitute urea compound is characterized in that, wherein R
1Be fluorenyl, naphthyl, phenyl or 3-chloro-4-fluorophenyl.
6. N-as claimed in claim 5 (2,6-benzyloxy benzoyl-)-N '-substitute urea compound is characterized in that; Described N-(2; 6-benzyloxy benzoyl-)-N '-substitute urea compound specifically: N-(2,6-benzyloxy benzoyl-)-N '-fluorenyl-urea, N-(2,6-benzyloxy benzoyl-)-N '-naphthyl-urea, N-(2; 6-benzyloxy benzoyl-)-N '-phenyl-urea or N-(2,6-benzyloxy benzoyl-)-N '-(3-chloro-4-fluorophenyl)-urea.
7. N-as claimed in claim 2 (2,6-benzyloxy benzoyl-)-N '-substitute urea compound is characterized in that; Described N-(2; 6-benzyloxy benzoyl-) N '-substitute urea compound specifically: N-(2,6-benzyloxy benzoyl-) N '-(1-xanthenyl)-urea, N-(2,6-benzyloxy benzoyl-)-N '-cyclohexyl-urea, N-(2; 6-benzyloxy benzoyl-)-N '-(2; 3-dihydro-indenes-1-yl)-urea, N-(2,6-benzyloxy benzoyl-)-N '-phenmethyl-urea, N-(2,6-benzyloxy benzoyl-) N '-styroyl-urea, N-(2; 6-benzyloxy benzoyl-)-N '-(4-fluorophenyl)-urea or N-(2,6-benzyloxy benzoyl-)-N '-diphenyl-methyl-urea.
8. a method for preparing like any described N-in the claim 1～7 (2,6-benzyloxy benzoyl-)-N '-substitute urea compound is characterized in that, the said method of said method comprises step (i), (ii) and (iii):
(i) by 2,6-difluorobenzonilyile preparation 2, the step of 6-benzyloxy BM;
(ii) by R
1-NH
2Preparation R
1The step of-NCO; With
(iii) under reflux state, by 2,6-benzyloxy BM and R
1-NCO carries out addition reaction and gets target compound.
9. like any described N-(2 in the claim 1～7; 6-benzyloxy benzoyl-)-N '-substitute urea compound is in preparation cyclophilin A (Cyclophilin A, CypA) application in the medicine of the disease of agonist or preparation prevention or treatment CypA mediation.
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| CN103992245A (en) * | 2014-05-23 | 2014-08-20 | 华东理工大学 | Substituted urea compound and use thereof |
| CN104058996A (en) * | 2014-06-23 | 2014-09-24 | 清华大学 | Anti-hepatitis C virus compound, preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101880246A (en) * | 2009-05-05 | 2010-11-10 | 中国科学院上海药物研究所 | Substituted benzoyl urea compound and preparation method and application thereof |
-
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101880246A (en) * | 2009-05-05 | 2010-11-10 | 中国科学院上海药物研究所 | Substituted benzoyl urea compound and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| SHUAISHUAI NI, ET AL.: "Discovering Potent Small Molecule Inhibitors of Cyclophilin A Using de Novo Drug Design Approach", 《JOURNAL OF MEDICINAL CHEMISTRY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103992245A (en) * | 2014-05-23 | 2014-08-20 | 华东理工大学 | Substituted urea compound and use thereof |
| CN104058996A (en) * | 2014-06-23 | 2014-09-24 | 清华大学 | Anti-hepatitis C virus compound, preparation method and application thereof |
| CN104058996B (en) * | 2014-06-23 | 2016-01-20 | 清华大学 | Anti-hepatitis c virus compound and its preparation method and application |
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