CN109651199A - A kind of histon deacetylase (HDAC) and the double target spot inhibitor of micro-pipe and preparation method thereof - Google Patents
A kind of histon deacetylase (HDAC) and the double target spot inhibitor of micro-pipe and preparation method thereof Download PDFInfo
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- CN109651199A CN109651199A CN201910016443.2A CN201910016443A CN109651199A CN 109651199 A CN109651199 A CN 109651199A CN 201910016443 A CN201910016443 A CN 201910016443A CN 109651199 A CN109651199 A CN 109651199A
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- silica gel
- gel column
- column chromatography
- ethyl acetate
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- 238000002360 preparation method Methods 0.000 title claims abstract description 23
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- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 197
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 120
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- YOHYSYJDKVYCJI-UHFFFAOYSA-N n-[3-[[6-[3-(trifluoromethyl)anilino]pyrimidin-4-yl]amino]phenyl]cyclopropanecarboxamide Chemical compound FC(F)(F)C1=CC=CC(NC=2N=CN=C(NC=3C=C(NC(=O)C4CC4)C=CC=3)C=2)=C1 YOHYSYJDKVYCJI-UHFFFAOYSA-N 0.000 description 1
- 210000001623 nucleosome Anatomy 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000031877 prophase Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- CHQMHPLRPQMAMX-UHFFFAOYSA-L sodium persulfate Substances [Na+].[Na+].[O-]S(=O)(=O)OOS([O-])(=O)=O CHQMHPLRPQMAMX-UHFFFAOYSA-L 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 230000003319 supportive effect Effects 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 229930185603 trichostatin Natural products 0.000 description 1
- RTKIYFITIVXBLE-QEQCGCAPSA-N trichostatin A Chemical compound ONC(=O)/C=C/C(/C)=C/[C@@H](C)C(=O)C1=CC=C(N(C)C)C=C1 RTKIYFITIVXBLE-QEQCGCAPSA-N 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C259/00—Compounds containing carboxyl groups, an oxygen atom of a carboxyl group being replaced by a nitrogen atom, this nitrogen atom being further bound to an oxygen atom and not being part of nitro or nitroso groups
- C07C259/04—Compounds containing carboxyl groups, an oxygen atom of a carboxyl group being replaced by a nitrogen atom, this nitrogen atom being further bound to an oxygen atom and not being part of nitro or nitroso groups without replacement of the other oxygen atom of the carboxyl group, e.g. hydroxamic acids
- C07C259/06—Compounds containing carboxyl groups, an oxygen atom of a carboxyl group being replaced by a nitrogen atom, this nitrogen atom being further bound to an oxygen atom and not being part of nitro or nitroso groups without replacement of the other oxygen atom of the carboxyl group, e.g. hydroxamic acids having carbon atoms of hydroxamic groups bound to hydrogen atoms or to acyclic carbon atoms
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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Abstract
本发明公开了一种组蛋白去乙酰化酶和微管双靶点抑制剂及其制备方法，该抑制剂如I)或II)所示：该抑制剂同时具有微管和组蛋白去乙酰化酶的双重抑制活性。
The invention discloses a dual-target inhibitor of histone deacetylase and microtubule and a preparation method thereof. The inhibitor is shown in I) or II): The inhibitor possesses dual inhibitory activity of microtubule and histone deacetylase at the same time.
Description
Technical field
The invention belongs to histon deacetylase (HDAC) inhibitor synthesis technical fields, specifically, being related to a kind of histone
Deacetylase and the double target spot inhibitor of micro-pipe and preparation method thereof.
Background technique
Histon deacetylase (HDAC) (histone deacetylase, HDAC) is the basic composition unit for maintaining chromosome
One of the crucial enzyme of acetylation of histone state balance in nucleosome, the deacetylation effect of catalysis histone, with gene
Transcription inhibition is closely related, involve promote gene silencing many processes, histone and nonhistones Acetylation status it is unbalance with
Generation, the development of tumour are closely related, therefore histon deacetylase (HDAC) is also the popular target in anti-tumor drug design.
Histon deacetylase (HDAC) inhibitor (histone deacetylase inhibitor, HDACi) is a kind of chemical combination
Object has the function of interference histon deacetylase (HDAC).HDACis is improved by the degree of acetylation of the intracellular histone of increase
The approach such as the isogenic expression of p21 inhibit the proliferation of tumour cell, induce tumor cell differentiation and (or) apoptosis.To swollen
Oncocyte migration, invasion, the inhibiting effect of transfer and Antineoplastic angiogenesis effect are also proved, therefore the exploitation of HDACi is
Research hotspot as neoplasm targeted therapy.
The pharmacophore of HDACi consists of three parts, cap structure, for identification the inlet of HDAC activity pocket；Zinc ion
Chelation group (ZBG), for chelating the zinc ion of HDAC catalytic pocket bottom；Connection chain, for connecting cap and ZBG, and with work
Property pocket in hydrophobic pathway interaction.And according to design feature, reported HDACi can be roughly divided into 4 major class: different
Hydroximic acid, benzamides, cyclic peptide and short-chain fat acids.Wherein hydroxamic acid include trichostatin and SAHA and
Its derivative etc..The research of this kind of hdac inhibitors attracts tremendous attention, it is proved to cause in low dose, low concentration
Tumor Differentiation, and selectively inhibit tumour growth and have no toxic side effect to normal cell, it is more suitable for clinic.Have 5 at present
Kind HDACis goes through listing for clinical (as shown in Figure 1).
The HDACs listed is undesirable to the therapeutic effect of solid tumor and with a variety of side effects etc., this serious limit
The application range of HDACi clinically is made.In order to solve this problem, the hdac inhibitor and design of double target spots are mentioned with synthesis
A kind of feasible scheme is supplied.Since the occurrence and development of tumour are related to multiple signal transduction pathway.Only inhibit an access
It is difficult to completely inhibit the proliferation of tumour.And studies have shown that the treatment effect of single target-point anti-cancer drug can be improved in multiple target-point anti-cancer drug
Fruit, and drug resistance is reduced, it is the important research direction of cancer drug development.The generation of histon deacetylase (HDAC) and tumour is close
Correlation, inhibitor can reduce the threshold value of apoptosis of tumor cells, have an extensive anti-tumor activity, and can with it is a variety of anti-swollen
Performance synergistic effect is used in combination in tumor medicine.This provides theoretical basis for the design and development of double target spot hdac inhibitors.Mesh
It is preceding to be in clinical investigation phase there are two multiple target point hdac inhibitor.And others HDAC multiple target point inhibitor such as IMPDH-
The bis- target spot inhibitor of HDAC, the double target spot inhibitor of II-HDAC of Topo, the bis- target spot inhibitor of EGFR/HER2-HDAC and HMGA-
Bis- target spot inhibitor of HDAC etc. also all show preferable anti-tumor activity.
Micro-pipe plays a supportive role in cell as important cytoskeleton, in prophase of cell division, microtubule depolymerization recombination
At spindle, while micro-pipe also participates in the signal transduction and migration of cell.Inhibit micro-pipe be proved to can blocks tumor cells have silk
Division and cycle progression, inducing apoptosis of tumour cell destroy tumor vessel and inhibit angiogenesis.Therefore micro-pipe is considered anti-
The important target spot of tumour medicine research and development.
Studies have shown that colchicin has the function of inhibiting tubulin polymerization.Colchicin and hdac inhibitor are joined
It closes in use, the two has synergistic antitumor effect.It is inspired by this, Zhang etc. is by introducing SAHA in colchicin molecule
Zinc Ions Chelated group of the tail groups as hdac inhibitor, synthesized the compound of a series of novel.Result of study table
These bright compounds not only have HDAC inhibiting effect, meanwhile, these compounds have the G2/M phase of BEL-7401 cell cycle
It is obviously prolonged effect, showing such compound also plays the role of inhibiting tubulin polymerization.This kind of chemical combination is found in MTT experiment
Object is to people epidermis A431 cell, human pulmonary epithelial cells, colon cancer HCT-116 cell, MCF-7 Breast Cancer Cell, prostate
Cancer PC-3 cell has inhibitory activity.
Chalcone SD400 can effectively inhibit the assembling of tubulin, have antitumor activity.Researcher is
A series of chalcone analog has been synthesized, the experimental results showed that these analogs have strong cytotoxicity, it can be effective
Ground inhibits the proliferation of tumour, and wherein compound 2a and 2b is the microtubule polymerization inhibitor (as shown in Figure 2) registered, two chemical combination
IC of the object to kinds of tumors50Value has reached the rank of nanomole.Although micro-pipe is a kind of important anti-tumor drug research and development target spot,
But micro-pipe and the report of the bis- target spot inhibitor of HDAC are few.
Summary of the invention
In view of this, the present invention provides a kind of histon deacetylase (HDAC) and the double target spot inhibitor of micro-pipe and its preparation sides
Method and application.
In order to solve the above-mentioned technical problem, the invention discloses a kind of histon deacetylase (HDAC)s and the double target spots of micro-pipe to inhibit
Agent, such as I) or II) shown:
Optionally, the specific structure and title of described (I) are as follows:
1) entitled N- hydroxyl -7- (2- methoxyl group -5- (3- ketone group -3- (3,4,5- trimethoxyphenyl) -1- (E)-the third
Alkenyl) phenoxy group-heptamide, abbreviation WBL-1, chemical structure is as follows:
2) entitled N- hydroxyl -8- (2- methoxyl group -5- (3- ketone group -3- (3,4,5- trimethoxyphenyl) -1- (E)-the third
Alkenyl) phenoxy group-caprylamide, abbreviation WBL-2, chemical structure is as follows:
3) entitled N- hydroxyl -7- (2- methoxyl group -5- (2- methyl -3- ketone group -3- (3,4,5- trimethoxyphenyl) -1-
(E)-acrylic) phenoxy group-heptamide, abbreviation WBL-3, chemical structure is as follows:
4) entitled N- hydroxyl -8- (2- methoxyl group -5- (2- methyl -3- ketone group -3- (3,4,5- trimethoxyphenyl) -1-
(E)-acrylic) phenoxy group-caprylamide, abbreviation WBL-4, chemical structure is as follows:
The specific structure and title of (II) are as follows:
1) entitled N1Hydroxy-n7(2- methoxyl group -5- (3- ketone group -3- (3,4,5- trimethoxyphenyl) -1- (E)-the third
Alkenyl) phenyl-heptanedioyl amine, abbreviation WBL-5, chemical structure is as follows:
2) entitled N1Hydroxy-n8(2- methoxyl group -5- (3- ketone group -3- (3,4,5- trimethoxyphenyl) -1- (E)-the third
Alkenyl) phenyl-suberamide, abbreviation WBL-6, chemical structure is as follows:
3) entitled N1Hydroxy-n7(2- methoxyl group -5- (2- methyl -3- ketone group -3- (3,4,5- trimethoxyphenyl) -
1- (E)-acrylic) phenyl-heptanedioyl amine, abbreviation WBL-7, chemical structure is as follows:
4) entitled N1Hydroxy-n8(2- methoxyl group -5- (2- methyl -3- ketone group -3- (3,4,5- trimethoxyphenyl) -
1- (E)-acrylic) phenyl-suberamide, abbreviation WBL-8, chemical structure is as follows:
The invention also discloses the preparation methods of a kind of histon deacetylase (HDAC) and the double target spot inhibitor of micro-pipe, including with
Lower step:
Step 1, synthesis A or B: weighing 3 ', 4 ', 5 '-trimethoxy acetophenones or 3 ', 4 ', 5 '-trimethoxy propiophenones with
3- hydroxyl -4-methoxybenzaldehyde is dissolved in methanol solvate, and the sodium hydroxide solution of 50% (W/V) is added to said mixture
In, mixture is stayed overnight under nitrogen protection；Hydrochloric acid is added to adjust the PH to 1 of mixture, methylene chloride is extracted twice, and TLC checks anti-
Answer situation；Anhydrous sodium sulfate dried filtrate, silica gel column chromatography purifies to obtain yellow solid A or B after concentration；
Step 2, synthesis AmES or BmES: A or B and m- bromine X acid methyl esters are dissolved in DMF, the Carbon Dioxide of doubling dose is added
Potassium, 60 DEG C of oil bath overnights, TLC are checked；Water and ethyl acetate is added to extract, anhydrous sodium sulfate dried filtrate, silicagel column color after concentration
Spectrometry purifies to obtain yellow solid AmES or BmES；
Step 3, synthesis AmAC or BmAC: AmES or BmES is mixed with 1N lithium hydroxide the water for being dissolved in volume ratio 1:1 and
In THF；25 DEG C of oil bath overnights, TLC check response situation, and adjusting PH is acidity, add EA to extract, anhydrous sodium sulfate dried filtrate is dense
Silica gel column chromatography purifies to obtain yellow solid AmAC or BmAC after contracting；
Step 4, synthesis AmTHP or BmTHP: by AmAC or BmAC, DCC and NH2OTHP is dissolved in DCM, 25 DEG C of oil bath overnights,
TLC checks response situation；Silica gel column chromatography purifies to obtain yellow solid AmTHP or BmTHP after filtering away DCC concentration；
Step 5, synthesis (I): being dissolved in methanol for AmTHP or BmTHP and CSA, and 68 DEG C of oil baths 3h, TLC check response situation,
Wherein, DCM:MEOH=20:1；HCl is adjusted to acidity, adds water, and EA extracts anhydrous sodium sulfate dried filtrate, silicagel column color after concentration
Spectrometry purifies to obtain yellow solid (I).
Optionally, 3 ', 4 ', the 5 '-trimethoxy acetophenones or 3 ' in the step 1,4 ', 5 '-trimethoxy propiophenones
It is 1:1 with 3- hydroxyl -4-methoxybenzaldehyde molar ratio, the PE:EA=2:1 in TLC inspection；In silica gel column chromatography purifying
PE:EA=4:1；The mole ratio of A or B and m- bromine X acid methyl esters in the step 2 are 1:1.1, and m is 7 or 8；X is heptan
Or it is pungent；PE:EA=2:1 in TLC inspection；PE:EA=6:1 in silica gel column chromatography purifying.
Optionally, the molar ratio of the AmES in the step 3 or BmES and 1N lithium hydroxide is 1:1.5；During TLC is checked
PE:EA=1:1；PE:EA=1:1 in silica gel column chromatography purifying；AmAC or BmAC:DCC in the step 4:
NH2OTHP=1mol:1.2mol:1.2mol；DCM:EA=3:1 in TLC inspection；DCM:EA in silica gel column chromatography purifying
=8:1.
Optionally, the AmTHP or BmTHP:CSA=4.8mol:1mol in the step 5；In silica gel column chromatography purifying
DCM:MEOH=100:1.
The invention also discloses the preparation methods of a kind of histon deacetylase (HDAC) and the double target spot inhibitor of micro-pipe, including with
Lower step:
Step 1, synthesis C or E: weighing 3 ', 4 ', 5 '-trimethoxy acetophenones or 3 ', 4 ', 5 '-trimethoxy propiophenones and
3- hydroxyl -4- nitrobenzaldehyde is dissolved in methanol solvate, and the potassium hydroxide solution of 50% (W/V) is added to said mixture
In, mixture is stayed overnight under nitrogen protection.TCL checks response situation, after the reaction was completed, hydrochloric acid is added to adjust the PH to 1 of mixture,
Methylene chloride is extracted twice；Anhydrous sodium sulfate dried filtrate, silica gel column chromatography purifies to obtain yellow solid C or E after concentration；
Step 2, synthesis D or F: nitro compound C or E are suspended in 100 milliliters of ethyl alcohol, are added 10 milliliters of distilled water, then plus
Enter reduced iron powder and ammonium chloride；The mixture is again return stirring 4 hours at 90 DEG C；It is down to room temperature, removes ethyl alcohol under vacuum condition,
100 milliliters of distilled water are added, pH value is adjusted to alkalescent (8-9) with 0.1M sodium hydrate aqueous solution；It is extracted with ethyl acetate
Three times, ethyl acetate layer is dry with anhydrous sodium sulfate；Crude product crosses silica gel column purification；Obtain yellow sterling D or F；
Step 3, synthesis CmES or DmES: amino-compound D or F are dissolved in 10 milliliters of anhydrous DMFs, and the X bis- of one times of amount is added
Sour mono-methyl is added with stirring the DIPEA of doubling dose HATU and four times of amounts；Mixture is stirred overnight at room temperature, and saturation is added
Saline solution is extracted with ethyl acetate three times, and ethyl acetate layer is dry with anhydrous sodium sulfate, and crude product uses column chromatographic purifying, obtains
Sterling CmES or DmES；
Step 4, synthesis CmAC or DmAC:CmES or DmES and 1N lithium hydroxide are dissolved in the water and THF of volume ratio 1:1；
25 DEG C of oil bath overnights, TCL check reaction process, and adjusting PH is acidity, and EA is added to extract, anhydrous sodium sulfate dried filtrate, silicon after concentration
Glue is column chromatography eluting to obtain yellow solid CmAC or DmAC:；
Step 5, synthesis CmTHP or DmTHP: by CmAC or DmAC, DCC and NH2OTHP is dissolved in DCM, 25 DEG C of oil bath overnights,
TCL checks response situation；Silica gel column chromatography purifies to obtain yellow solid CmTHP or DmTHP after filtering away DCC concentration；
Step 6, synthesis (II): CmTHP or DmTHP (0.9g, 1.6mmol): CSA=4.8:1mol, are dissolved in methanol, and 68 DEG C
Oil bath 3H, TCL check response situation；HCL is adjusted to acidity, adds water, and EA extracts anhydrous sodium sulfate dried filtrate, silicagel column after concentration
Chromatography purifies to obtain brown solid.
Optionally, 3 ', 4 ', 5 '-trimethoxy acetophenones in the step 1 and 3- hydroxyl -4-methoxybenzaldehyde
Molar ratio is 1:1, the PE:EA=2:1 in TLC inspection；PE:EA=4:1 in silica gel column chromatography purifying；In the step 2
Nitro compound, reduced iron powder and ammonium chloride molar ratio be 1:1:5；The mobile phase of silica gel column purification is ethyl acetate: stone
Oily ether=1:2.
Optionally, the m in the step 3 is 7 or 8, and X is heptan or pungent；The mobile phase of silica gel column purification is ethyl acetate: stone
Oily ether=1:3；The molar ratio of CmES or DmES and 1N lithium hydroxide in step 4 are 1:1.5；PE:EA=in TLC inspection
1:1；PE:EA=1:1 in silica gel column chromatography purifying.
Optionally, the CmAC or DmAC:DCC:NH in the step 52OTHP=1mol:1.2mol:1.2mol；TLC inspection
DCM:EA=3:1 in looking into；DCM:EA=8:1 in silica gel column chromatography purifying；CmTHP or DmTHP in the step 6:
CSA=4.8mol:1mol；DCM:MEOH=20:1 in TLC inspection；DCM:MEOH=100 in silica gel column chromatography purifying:
1。
Compared with prior art, the present invention can be obtained including following technical effect:
1) present invention has synthesized the Novel histone deacetylase inhibitor with chalcone skeleton structure for the first time.
2) compound that the present invention designs and synthesizes also has microtubule polymerization inhibitory activity.
3) wherein compound WBL-1, WBL-4, WBL-8 have strong suppression to the kinds of tumor cells including solid tumor
Production is used.
Certainly, it implements any of the products of the present invention it is not absolutely required to while reaching all the above technical effect.
Detailed description of the invention
The drawings described herein are used to provide a further understanding of the present invention, constitutes a part of the invention, this hair
Bright illustrative embodiments and their description are used to explain the present invention, and are not constituted improper limitations of the present invention.In the accompanying drawings:
Fig. 1 is 5 listed the hdac inhibitor in background of invention；
Fig. 2 is the structure of the compound 2a and 2b in background of invention；
Fig. 3 is that single concentration of the compounds of this invention inhibits the result of mixing HDAC enzyme.
Specific embodiment
Carry out the embodiment that the present invention will be described in detail below in conjunction with embodiment, whereby to the present invention how application technology hand
Section solves technical problem and reaches the realization process of technical effect to fully understand and implement.
The preparation of embodiment 1WBL-1
The structural formula of WBL-1 are as follows:
Its chemical name are as follows: N- hydroxyl -7- (2- methoxyl group -5- (3- ketone group -3- (3,4,5- trimethoxyphenyl) -1- (E) -
Acrylic) phenoxy group-heptamide.
Its synthetic route are as follows:
Specific step is as follows for its synthetic method:
Step 1, synthesis A: 3 ', 4 ', 5 are weighed,-trimethoxy acetophenone (21.02g, 1mmol) and 3- hydroxyl -4- methoxy
Benzaldehyde (15.22g, 1mmol) is dissolved in 200mL methanol, and the sodium hydroxide solution 160mL of 50% (W/V) is added to
It states in mixture, mixture is stirred overnight at room temperature under nitrogen protection.Methanol is evaporated off, and adds the pH value of 1M hydrochloric acid adjusting mixture
Three times with 20 milliliters of methylene chloride extractions methylene chloride Xiang Bingyong anhydrous sodium sulfate dried filtrate, silicon after concentration are collected to 1-2
It is yellow solid that glue column chromatography eluting (petroleum ether: ethyl acetate=4:1), which obtains sterling A, and yield 52% surveys 122 DEG C of fusing point；
The mole ratio of step 2, synthesis A7ES:A (3.4g, 10mmol) and 7- bromine methyl heptanoate is 1:1.1, and the two is molten
In the anhydrous N ' of 20mL, the Anhydrous potassium carbonate of doubling dose is added in N ' dimethylformamide, and 60 DEG C of oil bath heatings are stirred overnight, TLC inspection
(PE:EA=2:1 (v/v)) is surveyed to fully reacting.It is extracted 3 times after adding 200mL water with 20 milliliters of ethyl acetate, anhydrous sodium sulfate is dry
Dry ethyl acetate phase, it is solid for yellow to obtain A7ES sterling for silica gel column chromatography purifying (petroleum ether: ethyl acetate=6:1) after concentration
Body, yield 67%；
The molar ratio of step 3, synthesis A7AC:A7ES (3.2g, 6.7mmol) and 1M lithium hydroxide aqueous solution is 1:1.5,
Mixture is dissolved in the water of 30 milliliters of volume ratio 1:2 and the in the mixed solvent of tetrahydrofuran.25 DEG C are stirred overnight at room temperature, and TLC is checked
Tetrahydrofuran is evaporated off in response situation (PE:EA=1:1 (v/v)), and tune pH value is 1-2, adds ethyl acetate to extract, anhydrous sodium sulfate
Dry ethyl acetate phase, it is yellow solid, yield that silica gel column chromatography purifying (PE:EA=1:1), which obtains A7AC sterling, after concentration
It is 68%.
Step 4, synthesis A7THP:A7AC (2.2g, 4.5mmol) are dissolved in anhydrous methylene chloride, are successively added the two of 9mmol
The NH of carbodicyclo hexylimide and 9mmol2OTHP, 25 DEG C are stirred overnight at room temperature, and TLC checks response situation (methylene chloride: acetic acid
Ethyl ester=3:1 (v/v)).After being filtered to remove reaction residues, concentration of reaction solution directly crosses silica gel column purification (methylene chloride: acetic acid
Ethyl ester=8:1) obtain A7THP sterling be yellow solid, yield 59%.
Step 5, synthesis WBL-1:A7THP (1.5g, 2.7mmol): camphoric acid=4.8:1 (mol/mol) is dissolved in 20ml first
In alcohol, 68 DEG C of oil baths are stirred at reflux 3 hours, and TLC checks response situation (methylene chloride: methanol=20:1 (v/v)).It is down to room
Wen Hou is evaporated off methanol and adds water, and ethyl acetate extraction, ethyl acetate phase is dry with anhydrous sodium sulfate, silica gel column chromatography after concentration
It is yellow solid, yield 22% that method purifying (methylene chloride: methanol=100:1), which obtains WBL-1 sterling,.
Product WBL-1 is yellow solid,1H NMR(500MHz,DMSO)δ10.33(s,1H),8.65(s,1H),7.73
(q, J=15.5Hz, 2H), 7.48 (d, J=1.8Hz, 1H), 7.45 (dd, J=8.4,1.8Hz, 1H), 7.39 (s, 2H), 7.02
(d, J=8.4Hz, 1H), 4.03 (t, J=6.5Hz, 2H), 3.89 (s, 6H), 3.82 (s, 3H), 3.76 (s, 3H), 1.95 (t, J
=7.3Hz, 2H), 1.77-1.65 (m, 2H), 1.56-1.47 (m, 2H), 1.47-1.37 (m, 2H), 1.31 (m, 2H)13C NMR
(126MHz,DMSO)δ188.42,169.56,153.32,151.92,148.77,144.85,142.28,133.79,128.00,
123.81,120.13,113.38,112.27,106.66,68.85,60.64,56.69,56.12,32.70,29.12,28.82,
25.78,25.56.[M+H]+=487.2208.
The preparation of embodiment 2WBL-2
The chemical formula of WBL-2 are as follows:
Its chemical name is: N- hydroxyl -8- (2- methoxyl group -5- (3- ketone group -3- (3,4,5- trimethoxyphenyl) -1-
(E)-acrylic) phenoxy group-caprylamide.
Its synthetic route are as follows:
Specific step is as follows for its synthetic method:
Step 1, synthesis A: with 1 step 1 of embodiment；
The mole ratio of step 2, synthesis A8ES:A (3.44g, 10mmol) and 8- bromine methyl caprylate is 1:1.1, the two
It is dissolved in the anhydrous N ' of 20mL, the Anhydrous potassium carbonate of doubling dose, 60 DEG C of oil bath overnights is added in N ' dimethylformamide, and TLC checks reflection
Situation (petroleum ether: ethyl acetate=2:1 (v/v)).Water and ethyl acetate is added to extract, the dry ethyl acetate phase of anhydrous sodium sulfate,
Silica gel column chromatography purifying (petroleum ether: ethyl acetate=6:1) obtains yellow solid, yield 73% after concentration.
The molar ratio of step 3, synthesis A8AC:A8ES (3.6g, 7.3mmol) and 1M lithium hydroxide is 1:1.5, mixture
It is dissolved in the water and tetrahydrofuran mixed solution of 30mL volume ratio 1:2.25 DEG C are stirred overnight at room temperature, and TLC checks response situation (stone
Oily ether: ethyl acetate=1:1), tune PH is 1-2, add 20mL ethyl acetate to extract 3 times, the dry ethyl acetate phase of anhydrous sodium sulfate,
It is yellow solid that silica gel column chromatography purifying (petroleum ether: ethyl acetate=1:1), which obtains A8AC sterling, after concentration, and yield is
63%.
Step 4, synthesis A8THP:A8AC (2.3g, 4.9mmol): dicyclohexylcarbodiimide: NH2OTHP=1:1.2:
1.2 (mol/mol), are dissolved in anhydrous methylene chloride, and 25 DEG C are stirred overnight at room temperature, and TLC checks response situation DCM:EA=3:1
(v/v)；Dereaction residue is filtered out, silica gel column chromatography purifying (methylene chloride: ethyl acetate=8:1) obtains after concentration
A8THP sterling is yellow solid, yield 54%；
Step 5, synthesis WBL-2:A8THP (1.5g, 2.6mmol): camphoric acid=4.8:1 (mol/mol) is dissolved in 15 milliliters
Methanol, 68 DEG C of oil baths are stirred 3 hours, and TLC checks response situation (methylene chloride: methanol=20:1 (v/v)).Add water, acetic acid second
Ester extracts 3 times, the dry ethyl acetate phase of anhydrous sodium sulfate, silica gel column chromatography purifying after concentration (methylene chloride: methanol=100:
1) obtaining WBL-2 sterling is yellow solid, yield 25%.
Product WBL-2 is yellow solid,1H NMR(500MHz,DMSO)δ10.32(s,1H),8.64(s,1H),7.73
(q, J=15.5Hz, 2H), 7.49 (d, J=2Hz, 1H), 7.45 (dd, J=10.5,2Hz, 1H), 7.40 (s, 2H), 7.03 (d,
J=8.4Hz, 1H), 4.04 (t, J=8.1Hz, 2H), 3.90 (s, 6H), 3.82 (s, 3H), 3.76 (s, 3H), 1.94 (t, J=
9.2Hz,2H),1.72(m,2H),1.50(m,2H),1.41(m,2H),1.33(m,2H),1.26(m,2H).13C NMR
(126MHz,DMSO)δ188.42,169.58,153.32,151.91,148.78,144.85,142.28,133.79,128.00,
123.80,120.11,113.31,112.25,106.65,68.86,60.64,56.68,56.11,32.71,29.16,29.01,
28.93,25.94,25.54.[M+H]+=501.2366.
The preparation of embodiment 3WBL-3
The chemical formula of WBL-3 are as follows:
Its chemical name is: N- hydroxyl -7- (2- methoxyl group -5- (2- methyl -3- ketone group -3- (3,4,5- trimethoxy-benzene
Base) -1- (E)-acrylic) phenoxy group-heptamide.
Its synthetic route are as follows:
Specific step is as follows for its synthetic method:
Step 1, synthesis B: comparable 3 ', 4 ', the 5 '-trimethoxy propiophenone (15g, 66mmol) of mole and 3- hydroxyl are weighed
Base -4-methoxybenzaldehyde (33g, 66mmol) is dissolved in methanol solvate, and the sodium hydroxide solution of 11mL 50% (W/V) is added
Enter into said mixture, mixture is stirred to react overnight under nitrogen protection, and TLC checks response situation (petroleum ether: acetic acid second
Ester=2:1 (v/v)).1M hydrochloric acid is added to adjust the PH to 1-2 of mixture, 100mL methylene chloride is extracted twice.Anhydrous sodium sulfate is dry
Dry methylene chloride phase, silica gel column chromatography purifying (petroleum ether: ethyl acetate=4:1) obtains brown color sterling solid after concentration,
Yield 64%；
The mole ratio of step 2, synthesis B7ES:B (3.5g, 10mmol) and 7- bromine methyl heptanoate is 1:1.1, and the two is molten
In anhydrous n,N-Dimethylformamide, the Anhydrous potassium carbonate of twice of mole is added, 60 DEG C of oil baths are stirred overnight, and TLC checks (stone
Oily ether: ethyl acetate=2:1 (v/v)).Water and ethyl acetate is added to extract, the dry ethyl acetate phase of anhydrous sodium sulfate, silicon after concentration
It is yellow solid, yield 76% that glue column chromatography eluting (petroleum ether: ethyl acetate=6:1), which obtains B7ES sterling,；
The molar ratio of step 3, synthesis B7AC:B7ES (3.7g, 7.5mmol) and 1N lithium hydroxide is 1:1.5, mixture
It is dissolved in the water and THF of 30ml volume ratio 1:2；25 DEG C are stirred overnight at room temperature, and TLC checks response situation (PE:EA=1:1), adjust
PH value adds the extraction of 30ml ethyl acetate three times, anhydrous sodium sulfate dries ethyl acetate phase, silica gel column chromatography after concentration to 1-2
It is yellow solid, yield 54% that purifying (P petroleum ether: ethyl acetate=1:1), which obtains B7AC sterling,；
Step 4, synthesis B7THP:B7AC (2.0g, 4mmol): dicyclohexylcarbodiimide: NH2OTHP=1:1.2:1.2
(mol/mol), it is dissolved in anhydrous methylene chloride, 25 DEG C are stirred overnight at room temperature, and TLC checks response situation (methylene chloride: ethyl acetate
=3:1 (v/v)).Dereaction residue is filtered out, silica gel column chromatography purifying (methylene chloride: ethyl acetate=8:1) obtains after concentration
It is yellow solid, yield 51% to B7THP sterling；
Step 5, synthesis WBL-3:B7THP (1.2g, 2mmol): camphoric acid=4.8:1 (mol/mol) is dissolved in methanol, and 68
DEG C oil bath is stirred to react 3 hours, and TLC checks response situation (methylene chloride: methanol=20:1 (v/v)).Add water, ethyl acetate extraction
It takes three times, the dry ethyl acetate phase of anhydrous sodium sulfate, silica gel column chromatography purifying (methylene chloride: methanol=100:1) after concentration
Obtaining WBL-3 sterling is brown solid, yield 9%.
Product WBL-3 is yellow solid,1H NMR(500MHz,DMSO)δ10.32(s,1H),8.64(s,1H),7.19–
7.07 (m, 3H), 7.03 (d, J=8.3Hz, 1H), 6.97 (s, 1H), 3.96 (t, J=6.4Hz, 2H), 3.81 (s, 6H), 3.80
(s, 3H), 3.76 (s, 3H), 2.19 (s, 3H), 1.94 (t, J=7.3Hz, 2H), 1.70 (m, 2H), 1.50 (m, 2H), 1.40
(m,2H),1.29(m,2H).13C NMR(126MHz,DMSO)δ197.89,169.54,152.94,150.13,148.30,
141.89,140.98,134.21,133.89,128.46,123.74,115.25,112.24,107.27,68.63,60.58,
56.48,56.05,32.68,29.09,28.81,25.73,25.54,15.04.[M+H]+=501.2364.
The preparation of embodiment 4WBL-4
Its chemical formula are as follows:
Its chemical name is: N- hydroxyl -8- (2- methoxyl group -5- (2- methyl -3- ketone group -3- (3,4,5- trimethoxy-benzene
Base) -1- (E)-acrylic) phenoxy group-caprylamide；
Its synthetic route are as follows:
Specific step is as follows for its synthetic method:
Step 1, synthesis B: with embodiment 3；
The mole ratio of step 2, synthesis B8ES:B (3.5g, 10mmol) and 8- bromine methyl caprylate is 1:1.1, and the two is molten
In anhydrous n,N-Dimethylformamide, the Anhydrous potassium carbonate of twice of mole is added, 60 DEG C of oil baths are stirred overnight, and TLC checks (stone
Oily ether: ethyl acetate=2:1 (v/v))；Water and ethyl acetate is added to extract, ethyl acetate phase is dry with anhydrous sodium sulfate, after concentration
It is yellow solid, yield 82% that silica gel column chromatography purifying (petroleum ether: ethyl acetate=6:1), which obtains B8ES sterling,；
The molar ratio of step 3, synthesis B8AC:B8ES (4g, 8mmol) and 1M lithium hydroxide is 1:1.5, and mixture is dissolved in
In the water and tetrahydrofuran of 30ml volume ratio 1:2；25 DEG C are stirred overnight at room temperature, and TLC checks response situation (petroleum ether: acetic acid second
Ester=1:1 (v/v)), it adjusts pH value to 1-2, adds ethyl acetate extraction 3 times, the dry ethyl acetate phase of anhydrous sodium sulfate, silicon after concentration
It is yellow solid, yield 45% that glue column chromatography eluting (petroleum ether: ethyl acetate=1:1), which obtains B8ES sterling,；
Step 4, synthesis B8THP:B8AC (1.7g, 3.5mmol): dicyclohexylcarbodiimide: NH2OTHP=1:1.2:
1.2 (mol/mol) are dissolved in anhydrous methylene chloride, and 25 DEG C are stirred overnight at room temperature, and TLC checks response situation (methylene chloride: acetic acid
Ethyl ester=3:1 (v/v)).DCC reaction residues are filtered away, silica gel column chromatography purifies (methylene chloride: ethyl acetate after concentration
=8:1) obtain B8THP sterling be yellow solid, yield 88%；
Step 5, synthesis WBL-4:B8THP (1.8g, 3mmol): camphoric acid=4.8:1 (mol/mol) is dissolved in methanol, and 68
DEG C oil bath is stirred 3 hours, and TLC checks response situation (methylene chloride: methanol=20:1).PH value is adjusted to 1-2 using 1M HCl,
It is added water, ethyl acetate extraction, the dry ethyl acetate phase of anhydrous sodium sulfate, silica gel column chromatography purifying after concentration (methylene chloride:
Methanol=100:1) obtain WBL-10 sterling be brown solid, yield 12%.
Product WBL-4 is yellow solid,1H NMR(500MHz,DMSO)δ10.31(s,1H),8.64(s,1H),7.15-
7.11 (m, 3H), 7.03 (d, J=8.3Hz, 1H), 6.97 (s, 1H), 3.96 (t, J=6.4Hz, 2H), 3.82 (s, 6H), 3.80
(s, 3H), 3.76 (s, 3H), 2.19 (s, 3H), 1.94 (t, J=7.3Hz, 2H), 1.69 (m, 2H), 1.49 (m, 2H), 1.39
(m,2H),1.42–1.21(m,6H).13C NMR(126MHz,DMSO)δ197.89,169.55,152.94,150.13,
148.31,141.89,140.98,134.21,133.89,128.46,123.72,115.25,112.24,107.27,68.66,
60.58,56.49,56.05,32.70,29.13,28.99,28.92,25.88,25.52,15.04.[M+H]+=515.2520.
The preparation of embodiment 5WBL-5
The chemical formula of WBL-5 are as follows:
Its chemical name is: N1Hydroxy-n7(2- methoxyl group -5- (3- ketone group -3- (3,4,5- trimethoxyphenyl) -1-
(E)-acrylic) phenyl-heptanedioyl amine；
Its synthetic route are as follows:
Specific step is as follows for its synthetic method:
Step 1, synthesis C: comparable 3 ', 4 ', the 5 '-trimethoxy acetophenone (21g, 100mmol) of mole and 3- are weighed
Hydroxyl -4- nitrobenzaldehyde (15g, 100mmol) is dissolved in 200ml methanol, and the sodium hydroxide solution of 8ml 50% (W/V) is added
Enter into said mixture, mixture is stayed overnight under nitrogen protection.TCL checks response situation (petroleum ether: ethyl acetate=2:1
(v/v)) 1M HCl, after the reaction was completed, is added to adjust the PH to 1-2 of mixture, 100ml methylene chloride is extracted twice.Anhydrous slufuric acid
Sodium dry methylene chloride phase, silica gel column chromatography purifying (petroleum ether: ethyl acetate=4:1) obtains the sterling of C as Huang after concentration
Color solid, yield 62%；
Step 2, D: nitro compound C (18g, 100mmol) of synthesis are suspended in 200ml ethyl alcohol, and 20ml distilled water is added, then
Reduced iron powder (28g, 500mmol) and ammonium chloride (26g, 500mmol) is added；The mixture is again return stirring 4 hours at 90 DEG C；
Room temperature to be down to is evaporated off ethyl alcohol under vacuum condition, 200ml distilled water is added, by pH value 0.1M sodium hydrate aqueous solution tune
To alkalescent (8-9)；It is extracted with ethyl acetate three times, ethyl acetate layer is dry with anhydrous sodium sulfate.Crude product crosses silica gel column purification,
Mobile phase is ethyl acetate: petroleum ether=1:2.Obtain yellow sterling D, yield 78%；
Step 3, synthesis C7ES: amino-compound (2.5g, 7.5mmol) is dissolved in 10 milliliters of anhydrous N, N- dimethyl formyl
The pimelic acid mono-methyl of 1.2 equivalents is added in amine, is added with stirring the n,N-diisopropylethylamine of doubling dose HATU and four times of amounts；
Mixture is stirred overnight at room temperature, and saturated salt solution is added, is extracted with ethyl acetate three times, ethyl acetate layer anhydrous slufuric acid
Sodium is dry, and crude product uses column chromatographic purifying, and mobile phase is ethyl acetate: petroleum ether=1:3, obtains sterling, and yield is respectively
73%；
The molar ratio of step 4, synthesis C7AC:C7ES (2.7g, 5.4mmol) and 1M lithium hydroxide is 1:1.5, mixture
It is dissolved in the water and tetrahydrofuran of 30ml volume ratio 1:2；25 DEG C are stirred overnight at room temperature, and TCL checks reaction process (petroleum ether: second
Acetoacetic ester=1:1 (v/v)), adjusting PH is acidity, add ethyl acetate to extract, the dry ethyl acetate phase of anhydrous sodium sulfate, silicon after concentration
It is yellow solid, yield 61% that glue column chromatography eluting (petroleum ether: ethyl acetate=1:1), which obtains C7AC sterling,；
Step 5, synthesis C7THP:C7AC (1.5g, 3.2mmol): dicyclohexylcarbodiimide: NH2OTHP=1:1.2:
1.2 (mol/mol) is dissolved in anhydrous methylene chloride, 25 DEG C are stirred overnight at room temperature, and TCL checks response situation (methylene chloride: acetic acid
Ethyl ester=3:1 (v/v)).Filter out dereaction residue, after concentration silica gel column chromatography purifying (methylene chloride: ethyl acetate=8:
1) obtaining C7THP sterling is yellow solid, yield 50%；
Step 6, synthesis WBL-5:C7THP (0.9g, 1.6mmol): CSA=4.8:1mol, are dissolved in methanol, 68 DEG C of oil baths
3H, TCL check response situation (DCM:MEOH=20:1).Solution is adjusted to acidity with 1M HCl, adds water, ethyl acetate extracts,
Anhydrous sodium sulfate dries ethyl acetate phase, and silica gel column chromatography purifying (methylene chloride: methanol=100:1) obtains WBL- after concentration
5 sterlings are brown solid, yield 17%.
Product WBL-5 is yellow-brown solid,1H NMR(500MHz,DMSO)δ10.35(s,1H),8.66(s,1H),7.72
(q, J=15.5Hz, 2H), 7.52 (d, J=2.0Hz, 1H), 7.46 (dd, J=8.4,2.0Hz, 1H), 7.06 (s, 2H), 6.98
(d, J=8.4Hz, 1H), 3.89 (s, 6H), 3.82 (s, 3H), 3.76 (s, 3H), 2.36 (t, J=7.3Hz, 2H), 1.96 (t, J
=7.3Hz, 2H), 1.62 (m, 2H), 1.49 (m, 2H), 1.29 (m, 2H)13C NMR(126MHz,DMSO)δ188.42,
179.81,169.56,153.32,153.92,138.77,134.85,130.28,128.79,125.80,121.81,119.13,
112.38,112.27,100.66,60.82,56.71,55.80,38.30,29.12,28.82,26.01,24.95.[M+H]+=
501.2235.
The preparation of embodiment 6WBL-6
Its chemical formula are as follows:
Its chemical name is: N1Hydroxy-n8(2- methoxyl group -5- (3- ketone group -3- (3,4,5- trimethoxyphenyl) -1-
(E)-acrylic) phenyl-suberamide；
Its synthetic route are as follows:
Specific step is as follows for its synthetic method:
Step 1, synthesis C: with embodiment 5；
Step 2, synthesis D: with embodiment 5；
Step 3, synthesis C8ES: amino-compound (2.5g, 7.5mmol) is dissolved in the anhydrous n,N-Dimethylformamide of 10ml,
The suberic acid mono-methyl of one times of amount is added, is added with stirring the n,N-diisopropylethylamine of doubling dose HATU and four times of amounts.Mixing
Object is stirred overnight at room temperature, and saturated salt solution is added, is extracted with ethyl acetate three times, and ethyl acetate layer is dry with anhydrous sodium sulfate
Dry, crude product uses column chromatographic purifying, and mobile phase is ethyl acetate: petroleum ether=1:3, obtains sterling C8ES, and yield is respectively
78%；
The molar ratio of step 4, synthesis C8AC:C8ES (2.9g, 5.8mmol) and 1M lithium hydroxide is 1:1.5, mixture
It is dissolved in the water and tetrahydrofuran of 30ml volume ratio 1:2.25 DEG C are stirred overnight at room temperature, and TCL checks reaction process (petroleum ether: second
Acetoacetic ester=1:1 (v/v)), after tetrahydrofuran is evaporated off, adjusting solution PH is acidity, and ethyl acetate is added to extract, and anhydrous sodium sulfate is dry
Ethyl acetate phase, it is yellow solid that silica gel column chromatography purifying (petroleum ether: ethyl acetate=1:1), which obtains C8AC, after concentration, is produced
Rate is 66%；
Step 5, synthesis C8THP:C8AC (1.7g, 3.8mmol): dicyclohexylcarbodiimide: NH2OTHP=1:1.2:
1.2 (mol/mol) are dissolved in 50ml anhydrous methylene chloride, and 25 DEG C are stirred overnight at room temperature, TCL inspection response situation (methylene chloride:
Ethyl acetate=3:1 (v/v)).Filtering removal reaction residues, silica gel column chromatography purifies (methylene chloride: ethyl acetate after concentration
=8:1) obtain C8THP sterling be yellow solid, yield 56%；
Step 6, synthesis WBL-6:C8THP (1.2g, 2.1mmol): camphoric acid=4.8:1 (mol/mol) is dissolved in methanol,
68 DEG C of oil baths are stirred 3 hours, and TCL checks response situation (methylene chloride: methanol=20:1 (v/v)).Add water and methanol is evaporated off, second
Acetoacetic ester extracts three times, and the dry ethyl acetate phase of anhydrous sodium sulfate, silica gel column chromatography purifies (methylene chloride: methanol after concentration
=100:1), obtaining WBL-6 is brown solid, yield 19%.
Product WBL-6 is brown solid,1H NMR(500MHz,DMSO)δ10.36(s,1H),8.67(s,1H),7.73
(q, J=15.5Hz, 2H), 7.52 (d, J=2.0Hz, 1H), 7.49 (dd, J=8.4,2.0Hz, 1H), 7.05 (s, 2H), 6.97
(d, J=8.4Hz, 1H), 3.89 (s, 6H), 3.82 (s, 3H), 3.76 (s, 3H), 2.37 (t, J=7.3Hz, 2H), 1.97 (t, J
=7.3Hz, 2H), 1.63 (m, 2H), 1.50 (m, 2H), 1.31 (m, 2H), 1.29 (m, 2H)13C NMR(126MHz,DMSO)δ
188.60,179.82,169.66,153.62,153.97,138.87,134.90,130.29,128.81,125.82,121.83,
119.16,112.40,112.31,100.67,60.81,56.72,55.81,38.32,29.11,28.83,27.99,26.01,
24.96.[M+H]+=515.2391.
The preparation of embodiment 7WBL-7
Its chemical formula are as follows:
Its chemical name is: N1Hydroxy-n7(2- methoxyl group -5- (2- methyl -3- ketone group -3- (3,4,5- trimethoxy-benzene
Base) -1- (E)-acrylic) phenyl-heptanedioyl amine；
Its synthetic route are as follows:
Specific step is as follows for its synthetic method:
Step 1, synthesis E: comparable 3 ', 4 ', the 5 '-trimethoxy propiophenone (22g, 100mmol) of mole and 3- are weighed
Hydroxyl -4- nitrobenzaldehyde (15g, 100mmol) is dissolved in methanol solvate, and the sodium hydroxide solution of 8g 50% (W/V) is added
Into said mixture, mixture is stirred overnight at room temperature under nitrogen protection.TCL checks response situation (methylene chloride: acetic acid second
Ester=2:1 (v/v)), after the reaction was completed, 1M HCl is added to adjust the PH to 1-2 of mixture, 200ml methylene chloride is extracted twice.Nothing
Aqueous sodium persulfate dry methylene chloride phase, silica gel column chromatography purifying (petroleum ether: ethyl acetate=4:1) obtains E sterling after concentration
For yellow solid, yield 65%；
Step 2, F: nitro compound E (18g, 100mmol) of synthesis are suspended in 300 milliliters of ethyl alcohol, and 20 milliliters of steamings are added
Distilled water adds reduced iron powder (28g, 500mmol) and ammonium chloride (26g, 500mmol)；The mixture flows back at 90 DEG C and stirs
It mixes 4 hours；It is cooled to room temperature, ethyl alcohol is evaporated off under vacuum condition, add 500 milliliters of distilled water, by pH value 0.1M hydroxide
Sodium water solution is adjusted to alkalescent (8-9)；It is extracted three times with 200ml ethyl acetate, ethyl acetate layer is dry with anhydrous sodium sulfate.Slightly
Product cross silica gel column purification, and mobile phase is ethyl acetate: petroleum ether=1:2.Obtaining F is yellow solid, yield 58%；
Step 3, D7ES: amino-compound F (2.5g, 7.5mmol) of synthesis are dissolved in 20 milliliters of anhydrous N, N- dimethyl formyl
The single-acid ethyl ester of one times of amount is added in amine, is added with stirring the n,N-diisopropylethylamine of doubling dose HATU and four times of amounts.It is mixed
It closes object to be stirred overnight at room temperature, saturated salt solution is added, extracted three times with 20ml ethyl acetate, the anhydrous sulphur of ethyl acetate layer
Sour sodium is dry, and crude product uses column chromatographic purifying, and mobile phase is ethyl acetate: petroleum ether=1:3, obtains sterling, and yield is respectively
67%.
The molar ratio of step 4, synthesis D7AC:D7ES (2.5g, 5mmol) and 1M lithium hydroxide is 1:1.5, and mixture is molten
In the water and tetrahydrofuran of 30ml volume ratio 1:2.25 DEG C are stirred overnight at room temperature, and TCL checks reaction process (petroleum ether: acetic acid
Ethyl ester=1:1 (v/v)), tetrahydrofuran is evaporated off and adjusts the PH of solution to acidity, adds ethyl acetate extraction three times, anhydrous sodium sulfate
Dry ethyl acetate phase, it is yellow that silica gel column chromatography purifying (petroleum ether: ethyl acetate=1:1), which obtains D7AC sterling, after concentration
Solid, yield 57%；
Step 5, synthesis D7THP:D7AC (1.3g, 2.8mmol): dicyclohexylcarbodiimide: NH2OTHP=1:1.2:
1.2 (mol/mol) are dissolved in anhydrous methylene chloride, and 25 DEG C are stirred overnight at room temperature, and TCL checks response situation (methylene chloride: acetic acid
Ethyl ester=3:1 (v/v)).Filter out dereaction residue, after concentration silica gel column chromatography purifying (methylene chloride: ethyl acetate=8:
1) yellow solid, yield 58% are obtained；
Step 6, synthesis WBL-7:D7THP (0.9g, 1.6mmol): camphoric acid=4.8:1 (mol/mol) is dissolved in methanol,
68 DEG C of oil baths are stirred 3 hours, and TCL checks response situation (methylene chloride: methanol=20:1).Add water and methanol is evaporated off, acetic acid second
Ester extraction, the dry ethyl acetate phase of anhydrous sodium sulfate, silica gel column chromatography purifying (methylene chloride: methanol=100:1) after concentration
Obtaining WBL-7 sterling is brown solid, yield 13%.
Product WBL-7 is brown solid,1H NMR(500MHz,DMSO)δ10.39(s,1H),8.66(s,1H),7.61–
7.49 (m, 3H), 7.04 (s, 2H), 6.97 (d, J=8.4Hz, 1H), 3.81 (s, 6H), 3.80 (s, 3H), 3.76 (s, 3H),
2.35 (t, J=7.3Hz, 2H), 2.19 (s, 3H), 1.96 (t, J=7.3Hz, 2H), 1.65 (m, 2H), 1.51 (m, 2H), 1.29
(m,2H).13C NMR(126MHz,DMSO)δ190.89,179.82,169.90,153.66,149.13,146.31,140.89,
134.21,133.89,127.46,126.70,125.72,117.25,114.24,100.27,60.86,56.51,55.89,
38.33,32.56,27.76,25.11,24.91,15.01.[M+H]+=515.2392.
The preparation of embodiment 8WBL-8
The chemical formula of WBL-8 are as follows:
Its chemical name is: N1Hydroxy-n8(2- methoxyl group -5- (2- methyl -3- ketone group -3- (3,4,5- trimethoxy-benzene
Base) -1- (E)-acrylic) phenyl-suberamide；
Its synthetic route are as follows:
Specific step is as follows for its synthetic method:
Step 1, synthesis E: with embodiment 7；
Step 2, synthesis F: with embodiment 7；
Step 3, synthesis D8ES: amino-compound (2.5g, 7.5mmol) is dissolved in 10 milliliters of anhydrous DMFs, and one times of amount is added
Monomethyl succinate is added with stirring the DIPEA of doubling dose HATU and four times of amounts.Mixture is stirred overnight at room temperature, is added
Saturated salt solution is extracted with ethyl acetate three times, and ethyl acetate layer is dry with anhydrous sodium sulfate, and crude product uses column chromatographic purifying,
Mobile phase is ethyl acetate: petroleum ether=1:3, obtains sterling, and yield is respectively 78%；
The molar ratio of step 4, synthesis D8AC:D8ES (3.8g, 7.2mmol) and 1N lithium hydroxide is 1:1.5, mixture
It is dissolved in the water and THF of volume ratio 1:1.25 DEG C of oil bath overnights, TCL check reaction process (PE:EA=1:1), and adjusting PH is acidity,
EA is added to extract, anhydrous sodium sulfate dried filtrate, silica gel column chromatography purifying (PE:EA=1:1) obtains yellow solid after concentration, produces
Rate is 50%；
Step 5, synthesis D8THP:D8AC (1.8g, 3.6mmol): dicyclohexylcarbodiimide: NH2OTHP=1:1.2:
1.2 (mol/mol) are dissolved in anhydrous methylene chloride, and 25 DEG C are stirred overnight at room temperature, and TLC checks response situation (methylene chloride: acetic acid
Ethyl ester=3:1 (v/v)).Filtering removal reaction residues, after concentration silica gel column chromatography purifying (methylene chloride: ethyl acetate=8:
1) obtaining D8THP sterling is yellow solid, yield 60%；
Step 6, synthesis WBL-8:D8THP (1.3g, 2.1mmol): camphorsulfonic acid=4.8:1 (mol/mol) is dissolved in first
Alcohol, 68 DEG C of oil baths are stirred to react 3 hours, and TLC checks response situation (methylene chloride: methanol=20:1).Add water and methanol be evaporated off,
Ethyl acetate extraction, the dry ethyl acetate phase of anhydrous sodium sulfate, silica gel column chromatography purifies (methylene chloride: acetic acid second after concentration
Ester=100:1) obtain WBL-8 sterling be brown solid, yield 17%.
Product WBL-8 is brown solid,1H NMR(500MHz,DMSO)δ10.39(s,1H),8.66(s,1H),7.61–
7.49 (m, 3H), 7.04 (s, 2H), 6.97 (d, J=8.4Hz, 1H), 3.81 (s, 6H), 3.80 (s, 3H), 3.76 (s, 3H),
2.35 (t, J=7.3Hz, 2H), 2.19 (s, 3H), 1.96 (t, J=7.3Hz, 2H), 1.65 (m, 2H), 1.51 (m, 2H), 1.29
(m,2H).13C NMR(126MHz,DMSO)δ190.90,179.83,169.88,153.67,149.12,146.32,140.90,
134.22,133.90,127.45,126.71,125.72,117.26,114.25,100.30,60.85,56.52,55.88,
38.34,32.58,27.86,27.76,25.12,24.92,15.02. [M+H]+=529.2550.
Illustrate technical effect of the invention below with reference to specific experimental data:
Fig. 3 is single concentration inhibition mixing HDAC enzyme of 8 compounds as a result, compound concentration is all 1 μM.SAHA is sun
Property comparison medicine, DMSO is solvent.The result proves that compound WBL-1 to WBL-8 is tentatively shown to HDAC mixed enzyme
Potent inhibitory activity, wherein compound WBL-4 to single agriculture of HDAC mixed enzyme crosses inhibiting rate also slightly better than control drug SAHA,
The activity of WBL-8 and SAHA are close.
Inhibitory activity (nM) of the 1:8 compound of table to HDAC1,6,8
| Compound name | HDAC1 | HDAC6 | HDAC8 |
| WBL-1 | 25.8 | 11.2 | >1000 |
| WBL-2 | 50.7 | 17.1 | >1000 |
| WBL-3 | 377.0 | 130.7 | >1000 |
| WBL-4 | 36.1 | 14.2 | >1000 |
| WBL-5 | 36.8 | 22.6 | >1000 |
| WBL-6 | 56.9 | 27.2 | >1000 |
| WBL-7 | 211.2 | 97.8 | >1000 |
| WBL-8 | 46.6 | 19.3 | >1000 |
| SAHA | 44.6 | 21.6 | 540.6 |
| ACY1215 | 73.0 | 8.0 | 126.0 |
Wherein, SAHA and ACY1215 is positive control drug, unit nM.
It is listed the result shows that 8 selected compounds have HDAC1 and 6 two kinds of hypotypes, especially HDAC6 hypotype in table 1
There is good selectivity, and activity there is no under 1 μM of concentration to HDAC8.It is therefore intended that 8 compounds to HDAC1 and
6 two kinds of hypotypes have preferable selectivity.
Inhibitory activity (μM) of the 2:8 compound of table to tubulin polymerization
Wherein, positive control is compound 2a, and unit is μM.
It is listed the result shows that equally also being shown to micro-pipe in preferable 8 compounds of HDAC enzyme inhibition activity in table 2
The inhibitory activity of polymerization.
Inhibitory activity (μM) of 38 compounds of table to 3 kinds of tumor cell proliferations
| Compound | HCT-116(IC50) | MDA-MB-231(IC50) | HL-60(IC50) |
| SAHA | 5.06 | 5.15 | 2.88 |
| 2a | 3.63 | 7.56 | 1.91 |
| WBL-1 | 7.73 | 3.98 | 1.33 |
| WBL-2 | >10 | >10 | 5.63 |
| WBL-3 | 8.92 | 6.68 | 3.66 |
| WBL-4 | 7.44 | 3.83 | 1.13 |
| WBL-5 | 8.22 | 7.72 | 4.36 |
| WBL-6 | 9.06 | 5.56 | 4.89 |
| WBL-7 | >10 | >10 | 5.17 |
| WBL-8 | 7.76 | 4.16 | 1.68 |
Wherein, positive control drug is SAHA and 2a, and unit is μM.
As can be seen from Table 3, tri- compounds of compound WBL-1, WBL-4 and WBL-8 are to MDA-MB-231 and HL-60
The inhibitory activity of two kinds of cancer cells is preferable, and than two kinds positive drugs SAHA and 2a of activity are much better, IC50Below 5 μM of value,
So these three compounds can cook the drug candidate of potential treatment cancer.
Claims (10)
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