CN102702120A - New compound o-hydroxyphenyl tetrazine dicarbonamide, preparation and application thereof - Google Patents
New compound o-hydroxyphenyl tetrazine dicarbonamide, preparation and application thereof Download PDFInfo
- Publication number
- CN102702120A CN102702120A CN2012101003648A CN201210100364A CN102702120A CN 102702120 A CN102702120 A CN 102702120A CN 2012101003648 A CN2012101003648 A CN 2012101003648A CN 201210100364 A CN201210100364 A CN 201210100364A CN 102702120 A CN102702120 A CN 102702120A
- Authority
- CN
- China
- Prior art keywords
- tetrazine
- dimethyl
- organic solvent
- diformamide
- formula
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a new compound, N1,N4-di(2-hydroxyphenyl)-3,6-dimethyl-1,2,4,5-tetrazine-1,4-dicarbonamide shown in formula (I), and preparation and application thereof. The preparation of the compound comprises the steps of using 3,6-dimethyl-1,6-dihydro-1,2,4,5-tetrazine and bis(trichloromethyl) carbonate as raw materials to react with 2-hydroxyaniline in an organic solvent in the presence of alkaline catalyst. The compound provided by the invention is applied for preparing medicines for treatment and prevention of human lung cancer. According to the invention, the compound is rational in design, easy in operation, low in production cost and easy to obtain raw materials, and is suitable for industrialized application, and a new choice for screening anti-tumor medicines is provided.
Description
(1) technical field
The present invention relates to a kind of new compound o-hydroxy-phenyl tetrazine diformamide (is N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide) and preparation method thereof, and the application of said compound in the medicine of preparation prevention or treatment tumor disease.
(2) background technology
Tetrazine kind compound has many better physical character, spectral quality and high reaction activity and high, and especially the tetrazine derivatives of some special constructions has tangible antiviral activity, anti-tumor activity, and can be used as agricultural chemicals and sterilant.For example at present existing two kinds (four mite piperazines and the fluorine mite piperazine) listing of agricultural chemicals, an existing kind (antitumour drug TM) listing of medicine.
1978, bibliographical information 3,6-hexichol alkynyl-six hydrogen-1,2,4, the 5-tetrazine has anti-tumor activity and (consults Eremeev, A.V.; Tikhomirova, D.A.; Tyusheva, V.A.; Liepins, F.Khim.Geterotsikl.Soedin, 1978,753.), this is 1,2,4, the 5-tetrazine kind compound is in the news first and has the potential anti-tumor activity.Afterwards, reported 1,2,4 of some ad hoc structures successively, the 5-tetrazine kind compound has anti-tumor activity, for example has 3 of anti-tumor activity, two (the 2-hydroxyls-5-chloro-phenyl-)-1,2,4 of 6-, and the 5-tetrazine (is consulted Rao, G.-W.; Hu, W.-X.Bioorg.Med.Chem.Lett.2006,16 (14), 3702.), 1-isobutyryl-3,6-phenylbenzene-1,4-dihydro-1,2,4, the 5-tetrazine (is consulted Rao, G.-W.; Hu, W.-X.Bioorg.Med.Chem.Lett.2005,15 (12), 3714.) and N
1, N
4-two (aminomethyl phenyl)-3,6-dimethyl--1,2,4,5-tetrazine-1, the 4-diformamide (is consulted Hu, W.-X.; Rao, G.-W.; Sun, Y.-Q.Bioorg.Med.Chem.Lett.2004,14 (5), 1177.) etc., but great majority 1,2,4,5-tetrazine compound does not have anti-tumor activity.
(3) summary of the invention
The object of the present invention is to provide a kind of new compound o-hydroxy-phenyl tetrazine diformamide and preparation method thereof, i.e. N with antitumour activity
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide and preparation method thereof is for the screening of antitumor drug provides new kind.This compound is under doses, and human lung carcinoma cell line A-549 is had certain inhibiting rate, and (it is to the IC of A-549 JEG-3
50Be 21.9 μ g/mL).The present invention is reasonable in design, and the preparation method is easy, easy handling, raw material be easy to get and production cost lower, be suitable for industrial applications.
The technical scheme that the present invention adopts is following:
A kind of new compound o-hydroxy-phenyl tetrazine diformamide provided by the invention, i.e. N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide compound, it has following structural (I):
N of the present invention
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1; The preparation method of 4-diformamide (I) is following: under-10～0 ℃ of condition, with 3 shown in the formula (II), 6-dimethyl--1,6-dihydro-1,2; 4, and 5-tetrazine, two (trichloromethyl) carbonic ether (TRIPHOSGENE 99.5, BTC), basic catalyst adds in the organic solvent A, after adding, heating reflux reaction 4～12 hours; Reaction cools to reaction solution about 5 ℃ after finishing, and normally cools to 4～6 ℃, leads to nitrogen, drips the solution of the organic solvent B of 2-hydroxyanilines again, after dripping off; Reflux 12～48 hours, reaction finishes, and after steaming desolventizes, residue recrystallization or column chromatography is obtained the N shown in the formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2; 4,5-tetrazine-1,4-diformamide compound; Described basic catalyst is one of following: pyridine, triethylamine, quinoline, Pottasium Hydroxide, sodium hydroxide, N, N-xylidene(s), 4-Dimethylamino pyridine or 4-pyrrolidyl pyridine;
Preparation N according to the invention
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the reaction of 4-diformamide (I) is shown in following reaction formula:
Of the present invention 3, the feed intake ratio of amount of substance of 6-dimethyl--1,6-dihydro-1,2,4,5-tetrazine (II) and two (trichloromethyl) carbonic ethers, basic catalyst, 2-hydroxyanilines is 1: 0.67～2: 0.1～1: 2～6.The TV consumption of said organic solvent A and organic solvent B is with shown in the formula (II) 3,6-dimethyl--1, and 6-dihydro-1,2,4,5-tetrazine quality is counted 13～75ml/g.
The present invention reacts used organic solvent A and organic solvent B and independently is selected from one of following separately: methylene dichloride, chloroform, toluene, methyl alcohol, ethanol or THF.
Concrete, the present invention prepares N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the method for 4-diformamide (I) recommends to carry out as follows: with shown in organic solvent A 1 dissolution type (II) 3; 6-dimethyl--1,6-dihydro-1,2,4,5-tetrazine and basic catalyst obtain containing 3; 6-dimethyl--1,6-dihydro-1,2,4, the solution of 5-tetrazine and basic catalyst places reactor drum with two (trichloromethyl) carbonic ethers and organic solvent A 2; Stir down at-10～0 ℃, slowly drip and describedly contain 3,6-dimethyl--1,6-dihydro-1,2; 4, the solution of 5-tetrazine and basic catalyst, 10～80min is reinforced to finish, reflux 4～12 hours, TLC follows the tracks of detection; Reaction cools to reaction solution about 5 ℃ after finishing, and normally drops to 4～6 ℃, and logical nitrogen drips the solution that the 2-hydroxyanilines is dissolved in organic solvent B, after dripping off again; Reflux 12～48 hours, TLC follows the tracks of detection, and reaction finishes, and after steaming desolventizes, residue recrystallization or column chromatography is obtained the N shown in the formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2; 4; 5-tetrazine-1,4-diformamide compound, described basic catalyst are one of following: pyridine, triethylamine, quinoline, Pottasium Hydroxide, sodium hydroxide, N; N-xylidene(s), 4-Dimethylamino pyridine or 4-pyrrolidyl pyridine; Described organic solvent A 1 is identical organic solvent with organic solvent A 2, and organic solvent B and organic solvent A 1 are identical or different organic solvent, organic solvent A 1, organic solvent A 2 be unified be selected from one of following: methylene dichloride, chloroform, toluene, methyl alcohol, ethanol or THF; It is one of following that described organic solvent B is selected from: methylene dichloride, chloroform, toluene, methyl alcohol, ethanol or THF; The TV consumption of said organic solvent A 1 and organic solvent A 2 is with shown in the formula (II) 3,6-dimethyl--1, and 6-dihydro-1,2,4,5-tetrazine quality is counted 10～50ml/g; The volumetric usage of said organic solvent B is with shown in the formula (II) 3,6-dimethyl--1,6-dihydro-1,2,4; 5-tetrazine quality is counted 3～25ml/g, and said organic solvent A 1 consumption is can dissolve 3,6-dimethyl--1,6-dihydro-1,2; 4,5-tetrazine and basic catalyst get final product, and be described 3,6-dimethyl--1; The feed intake ratio of amount of substance of 6-dihydro-1,2,4,5-tetrazine (II) and two (trichloromethyl) carbonic ethers, basic catalyst, 2-hydroxyanilines is 1: 0.67～2: 0.1～1: 2～6.
Residue recrystallization described in the method for the present invention is to carry out as follows: get the residue that steams after desolventizing in reaction flask, add recrystallization solvent, stirring heating; Refluxed 5～10 minutes; Filtered while hot is removed insolubles, and solid is separated out in the filtrating cooling; Refilter, the filter cake oven dry obtains the N shown in the formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide compound; Said recrystallization solvent is one of following: methyl alcohol, ethanol, sherwood oil, ETHYLE ACETATE, methylene dichloride, chloroform or THF are preferably methyl alcohol, ethanol, ETHYLE ACETATE, methylene dichloride, chloroform or THF.
Residue column chromatography described in the method for the present invention carries out as follows: get the residue that steams after desolventizing in the single port bottle, add organic solvent C it is dissolved, obtain lysate; In lysate, add the column chromatography silica gel of 1.5～2 times of amounts of residue quality then, behind the mixing, steaming desolventizes; The mixture of exsiccant residue and silica gel, mixture is adorned post, be that 0.5～10: 1 sherwood oil and ethyl acetate mixture are eluent with volume ratio then; Directly carry out wash-out, TLC follows the tracks of detection, detects according to TLC and collects the elutriant that contains the compound shown in the formula (I); Collection liquid is dry, the N shown in the acquisition formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide compound; Said organic solvent C is one of following: methyl alcohol, ethanol, ETHYLE ACETATE, methylene dichloride, chloroform, THF or DMSO 99.8MIN..
Organic solvent A of the present invention (organic solvent A 1 and organic solvent A 2), organic solvent B; The represented organic solvent that is used to react that all is meant; The letter does not here refer in particular to the implication of certain some organic solvent; Letter is just answered clear for the ease of table, be used for distinguishing these organic solvents and appear in the different reactions step.
N of the present invention
1, N
4-two (2-hydroxy phenyls)-3; 6-dimethyl--1,2,4; 5-tetrazine-1; 4-diformamide (I) can be applicable to prepare the medicine of prevention or treatment tumor disease, the particularly application in the medicine of preparation prevention or treatment people lung cancer disease, and described human lung carcinoma cell line is human lung carcinoma cell line A-549.
Compound provided by the invention is (The compounds of this invention concentration is 100 μ g/mL, 10 μ g/mL, 1 μ g/mL) under doses, and human lung carcinoma cell line A-549 is had certain inhibiting rate, and it is to the IC of A-549 JEG-3
50Be 21.9 μ g/mL; (The compounds of this invention concentration is 100 μ g/mL to compound according to the invention under doses; 10 μ g/mL; 1 μ g/mL) human breast cancer cell strain MCF-7 is also had certain inhibiting rate, and with the The compounds of this invention similar be that substrate synthetic compound not necessarily has antitumour activity or antitumour activity is lower with other amine.
Beneficial effect of the present invention is mainly reflected in: (1) the invention provides a kind of tetrazine compound novel, that have good antitumour activity, i.e. N shown in the formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide compound; For the screening of cancer drug provides new variety; (2) the invention provides the preparation method of this tetrazine compound, this preparation method is simple, easy handling, raw material be easy to get and production cost lower, be suitable for practicality, be expected to be applied to prepare in the medicine of prevention or treatment tumor disease.
(4) description of drawings
Compound N among Fig. 1 embodiment 9
1, N
4-two (4-ethoxyl phenenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (III), N
1, N
4-dimethyl--3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (IV) and N
1, N
4-two (3-nitrophenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the reaction formula schema of 4-diformamide (V).
(5) embodiment
The present invention combines specific embodiment to be further described, and following embodiment is that explanation is of the present invention, rather than limits the present invention by any way.
3,6-dimethyl--1,6-dihydro-1,2,4, preparation reference literature (Sun, the Y.Q. of 5-tetrazine (II); Hu, W.X.; Yuan, Q.Synth.Commun.2003,33,2769.) method prepare.
Embodiment 1:N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the preparation of 4-diformamide (I)
With 2.00 gram (17.8mmol) 3,6-dimethyl--1,6-dihydro-1,2; 4,5-tetrazine (II) and 2.16 gram (17.8mmol) N, the N-xylidene(s) is dissolved in 50 milliliters of dichloromethane solutions processes preparation solution, (promptly contains 3; 6-dimethyl--1,6-dihydro-1,2,4; The solution of 5-tetrazine and basic catalyst), add 20 milliliters of two (trichloromethyl) carbonic ether 10.60 grams (35.7mmol) and methylene dichloride in 250 milliliters of there-necked flasks successively, stir down, slowly drip preparation solution at 0 ℃.After adding in the 10min, TLC follows the tracks of detection, back flow reaction 4 hours.Be cooled to 5 ℃ then, logical nitrogen drips 3.90 gram (35.7mmol) 2-hydroxyanilines again and is dissolved in the solution that 20 milliliters of methylene dichloride obtain.After dripping off, TLC follows the tracks of detection, and reflux 48 hours is steamed and removed methylene dichloride; The residue column chromatography is promptly got the residue that steams after desolventizing and is added 20 ml methanol solvents it is dissolved, and obtains lysate, in lysate, adds 2.2 gram silica gel (300～400 order gross porosity (zcx.II) type column chromatography silica gel) then; Behind the mixing, steaming desolventizes, and gets the mixture of exsiccant residue and silica gel, and mixture is adorned post; Be eluent with 1: 2 sherwood oil of volume ratio and ethyl acetate mixture then, wash-out, TLC follows the tracks of detection, detects according to TLC and collects the elutriant that contains the compound shown in the formula (I); Collection liquid is dry, obtains white solid 0.34 gram, i.e. N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (I), yield 5.0% (with formula (II) 3,6-dimethyl--1,6-dihydro-1,2,4,5-tetrazine amount of substance meter, down together), 246～248 ℃ of fusing points.
1H?NMR(500MHz，CDCl
3)δ：2.42(s，6H，CH
3)，6.80-6.83(m，2H，C
6H
4)，6.89-6.95(m，4H，C
6H
4)，7.98(dd，2H，J＝1.2，6.8Hz，C
6H
4)，9.02(s，2H)，10.16(s，2H)。
Embodiment 2:N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the preparation of 4-diformamide (I)
With 2.00 gram (17.8mmol) 3,6-dimethyl--1,6-dihydro-1; 2; 4,5-tetrazine (II) and 0.98 gram (12.4mmol) pyridine are dissolved in processes preparation solution in 10 ml methanol, add 10 milliliters of two (trichloromethyl) carbonic ether 3.54 grams (11.9mmol) and methyl alcohol in 50 milliliters of there-necked flasks successively; Stir down at-5 ℃, slowly drip preparation solution.After adding in the 10min, TLC follows the tracks of detection, back flow reaction 10 hours.Be cooled to 6 ℃ then, logical nitrogen, Dropwise 5 .84 gram (53.5mmol) 2-hydroxyanilines is dissolved in the solution that 6 milliliters of ethanol obtain again.After dripping off, TLC follows the tracks of detection, and reflux 27 hours is steamed and removed methyl alcohol and ethanol, and residue adds 20 milliliters of chloroforms; Stirring heating, the 5～10min that refluxes, filtered while hot is removed insolubles, and solid is separated out in the filtrating cooling; Filter, 50 ℃ of oven dry of filter cake obtain white solid 0.78 gram, i.e. N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (I), yield 11.4%, 246～248 ℃ of fusing points.
1H NMR is with embodiment 1.
Embodiment 3:N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the preparation of 4-diformamide (I)
With 2.00 gram (17.8mmol) 3,6-dimethyl--1,6-dihydro-1; 2; 4,5-tetrazine (II) and 0.91 gram (7.4mmol) 4-Dimethylamino pyridine are dissolved in 40 milliliters of chloroformic solutions processes preparation solution, adds 20 milliliters of two (trichloromethyl) carbonic ether 4.30 grams (14.5mmol) and chloroforms in 100 milliliters of there-necked flasks successively; Stir down at-10 ℃, slowly drip preparation solution.After adding in the 40min, TLC follows the tracks of detection, back flow reaction 8 hours.Be cooled to 6 ℃ then, logical nitrogen drips 7.77 gram (71.2mmol) 2-hydroxyanilines again and is dissolved in the solution that 10 milliliters of chloroforms obtain.After dripping off, TLC follows the tracks of detection, and reflux 12 hours is steamed and removed chloroform, and residue is with 20 milliliters of ethyl alcohol recrystallizations, and other are operated with embodiment 2, obtains white solid 0.49 gram, i.e. N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (I), yield 7.2%, 246～248 ℃ of fusing points.
1H NMR is with embodiment 1.
Embodiment 4:N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the preparation of 4-diformamide (I)
With 2.00 gram (17.8mmol) 3,6-dimethyl--1,6-dihydro-1; 2; 4,5-tetrazine (II) and 1.75 gram (13.5mmol) quinoline are dissolved in 40 milliliters of tetrahydrofuran solutions processes preparation solution, adds 30 milliliters of two (trichloromethyl) carbonic ether 9.06 grams (30.5mmol) and THFs in 250 milliliters of there-necked flasks successively; Stir down at-8 ℃, slowly drip preparation solution.After adding in the 30min, TLC follows the tracks of detection, back flow reaction 12 hours.Be cooled to 4 ℃ then, logical nitrogen drips 9.72 gram (89.1mmol) 2-hydroxyanilines again and is dissolved in the solution that 30 milliliters of chloroforms obtain.After dripping off, TLC follows the tracks of detection, and reflux 28 hours is steamed and removed THF and chloroform, and residue is with 20 milliliters of re-crystallizing in ethyl acetate, and other are operated with embodiment 2, obtains white solid 1.39 gram, i.e. N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (I), yield 20.4%, 246～248 ℃ of fusing points.
1H NMR is with embodiment 1.
Embodiment 5:N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the preparation of 4-diformamide (I)
With 2.00 gram (17.8mmol) 3,6-dimethyl--1,6-dihydro-1; 2; 4,5-tetrazine (II) and 1.60 gram (10.8mmol) 4-pyrrolidyl pyridines are dissolved in 80 milliliters of ethanolic solns processes preparation solution, adds 20 milliliters of two (trichloromethyl) carbonic ether 8.00 grams (27.0mmol) and ethanol in 250 milliliters of there-necked flasks successively; Stir down at 0 ℃, slowly drip preparation solution.After adding in the 80min, TLC follows the tracks of detection, back flow reaction 6 hours.Be cooled to 5 ℃ then, logical nitrogen drips 11.67 gram (106.9mmol) 2-hydroxyanilines again and is dissolved in the solution that 40 milliliters of ethanol obtain.After dripping off, TLC follows the tracks of detection, and reflux 36 hours is steamed and removed ethanol, and residue is with 20 milliliters of THF recrystallizations, and other are operated with embodiment 2, obtains white solid 1.03 gram, i.e. N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (I).Yield 15.1%, 246～248 ℃ of fusing points.
1H NMR is with embodiment 1.
Embodiment 6:N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the preparation of 4-diformamide (I)
With 2.00 gram (17.8mmol) 3,6-dimethyl--1,6-dihydro-1; 2; 4,5-tetrazine (II) and 0.18 gram (1.78mmol) triethylamine are dissolved in 50 milliliters of toluene solutions processes preparation solution, adds 50 milliliters of two (trichloromethyl) carbonic ether 10.60 grams (35.7mmol) and toluene in 250 milliliters of there-necked flasks successively; Stir down at-10 ℃, slowly drip preparation solution.After adding in the 70min, TLC follows the tracks of detection, back flow reaction 12 hours.Be cooled to 5 ℃ then, logical nitrogen, Dropwise 5 .84 gram (53.5mmol) 2-hydroxyanilines is dissolved in the solution that 50 milliliters of toluene obtain again.After dripping off, TLC follows the tracks of detection, and reflux 18 hours is steamed and removed toluene, and residue is with 20 milliliters of methylene dichloride recrystallizations, and other are operated with embodiment 2, obtains white solid 1.22 gram, the i.e. N shown in the formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (I), yield 17.9%, 246～248 ℃ of fusing points.
1H NMR is with embodiment 1.
Embodiment 7:N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the preparation of 4-diformamide (I)
With 2.00 gram (17.8mmol) 3,6-dimethyl--1,6-dihydro-1; 2; 4,5-tetrazine (II) and 0.36 gram (9.0mmol) sodium hydroxide are dissolved in 50 milliliters of toluene solutions processes preparation solution, adds 50 milliliters of two (trichloromethyl) carbonic ether 10.60 grams (35.7mmol) and toluene in 250 milliliters of there-necked flasks successively; Stir down at-10 ℃, slowly drip preparation solution.After adding in the 70min, TLC follows the tracks of detection, back flow reaction 12 hours.Be cooled to 5 ℃ then, logical nitrogen, Dropwise 5 .84 gram (53.5mmol) 2-hydroxyanilines is dissolved in the solution that 50 milliliters of toluene obtain again.After dripping off, TLC follows the tracks of detection, and reflux 18 hours is steamed and removed toluene, and residue is with 20 ml methanol recrystallizations, and other are operated with embodiment 2, obtains white solid 0.22 gram, the i.e. N shown in the formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (I), yield 3.2%, 246～248 ℃ of fusing points.
1H NMR is with embodiment 1.
Embodiment 8:N
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the preparation of 4-diformamide (I)
With 2.00 gram (17.8mmol) 3,6-dimethyl--1,6-dihydro-1; 2; 4,5-tetrazine (II) and 0.20 gram (3.6mmol) Pottasium Hydroxide are dissolved in 40 milliliters of tetrahydrofuran solutions processes preparation solution, adds 30 milliliters of two (trichloromethyl) carbonic ether 9.06 grams (30.5mmol) and THFs in 250 milliliters of there-necked flasks successively; Stir down at-8 ℃, slowly drip preparation solution.After adding in the 30min, TLC follows the tracks of detection, back flow reaction 12 hours.Be cooled to 4 ℃ then, logical nitrogen drips 9.72 gram (89.1mmol) 2-hydroxyanilines again and is dissolved in the solution that 30 milliliters of chloroforms obtain.After dripping off, TLC follows the tracks of detection, reflux 29 hours; Steam and remove THF and chloroform, (eluent is a sherwood oil: ETHYLE ACETATE=10: 1 (volume ratio)), residue dissolves with 20 milliliters of methylene dichloride the residue column chromatography; Other operations obtain white solid 0.35 gram, i.e. N with embodiment 1
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (I), yield 5.1%, 246～248 ℃ of fusing points.
1H NMR is with embodiment 1.
Embodiment 9: the antitumour activity vitro test
1) compound that embodiment 1 is made (I) has carried out human breast carcinoma and human lung carcinoma cell line biological activity test.The result shows that compound (I) has the good anticancer activity to human lung carcinoma cell line A-549.
Testing method: tetrazolium (Methyl-Thiazol-Tetrozolium, MTT) reduction method
Cell strain: human breast cancer cell strain MCF-7 and human lung carcinoma cell line A-549
Action time: 72 hours
The compound (I) that embodiment 1 is made is with DMSO (DMSO 99.8MIN.) dissolving (consumption of DMSO with can dissolved compound get final product) configuration liquid concentrator, again with liquid concentrator with substratum (the DMEM substratum of Gibco company production; Culture medium preparation: contain 800,000 unit penicillium mould in every 1000mL substratum; 1.0g Streptomycin sulphate; 10% inactivated fetal bovine serum) is diluted to working concentration (100 μ g/mL; 10 μ g/mL and 1 μ g/mL), then human breast cancer cell strain MCF-7 and human lung carcinoma cell line A-549 are carried out the antitumour activity test respectively.The result of test is as shown in table 1 below:
Table 1 compound (I) is to the restraining effect of MCF-7 and A-549 growth of cancer cells
2) according to embodiment 1, the 2-hydroxyanilines is substituted with 4-phenetidine, methylamine or 3-N-methyl-p-nitroaniline respectively, other are operated with embodiment 1, have synthesized compound N respectively
1, N
4-two (4-ethoxyl phenenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (III), N
1, N
4-dimethyl--3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (IV) and N
1, N
4-two (3-nitrophenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide (V), the reaction formula flow process is as shown in Figure 1.
According to aforesaid method; With the compound that makes (III) and (IV) carried out human breast cancer cell strain biological activity test; Test result shows compound (III) and (IV) human breast cancer cell strain MCF-7 is suppressed effect all relatively poor; With the compound that makes (III), (IV) and (V) carried out the human lung carcinoma cell line biological activity test; Test result shows compound (III), (IV) and (V) human lung carcinoma cell line A-549 is suppressed effect all relatively poor, compound (III), (IV) and (V) antitumour activity of human lung carcinoma cell line A-549 can not show a candle to compound (I).Concrete outcome is as shown in table 2 below:
Table 2 compound is to the restraining effect of MCF-7 and A-549 growth of cancer cells
Above-mentioned experiment shows: compound (I) is remarkable to the restraining effect of human lung carcinoma cell line A-549 growth, and is not obvious to the restraining effect of human breast cancer cell strain MCF-7 growth.And the compound of other 2 similar (III) and (IV) all not obvious to the restraining effect of human breast cancer cell strain MCF-7 growth, the compound of other 3 similar (III), (IV) are all not obvious with the restraining effect of (V) human lung carcinoma cell line A-549 being grown.
Claims (9)
1. one kind suc as formula the N shown in (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide compound:
2. one kind prepares N as claimed in claim 1
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1; The method of 4-diformamide (I) is characterized in that described method is: under-10～0 ℃ of condition, with 3 shown in the formula (II), 6-dimethyl--1,6-dihydro-1; 2,4,5-tetrazine, two (trichloromethyl) carbonic ether, basic catalyst add in the organic solvent A, after adding, and heating reflux reaction 4～12 hours; Then reaction solution is cooled to 4～6 ℃, lead to nitrogen, drip the solution of the organic solvent B of 2-hydroxyanilines again, after dripping off; Reflux 12～48 hours, reaction finishes, and after steaming desolventizes, residue recrystallization or column chromatography is obtained the N shown in the formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2; 4,5-tetrazine-1,4-diformamide compound; Described basic catalyst is one of following: pyridine, triethylamine, quinoline, Pottasium Hydroxide, sodium hydroxide, N, N-xylidene(s), 4-Dimethylamino pyridine or 4-pyrrolidyl pyridine;
3. method as claimed in claim 2, it is characterized in that described 3,6-dimethyl--1,6-dihydro-1; 2,4, the feed intake ratio of amount of substance of 5-tetrazine (II) and two (trichloromethyl) carbonic ethers, basic catalyst, 2-hydroxyanilines is 1: 0.67～2: 0.1～1: 2～6; The TV consumption of said organic solvent A and organic solvent B is with shown in the formula (II) 3,6-dimethyl--1,6-dihydro-1; 2,4,5-tetrazine quality is counted 13～75ml/g.
4. method as claimed in claim 2, it is one of following to it is characterized in that described organic solvent A and organic solvent B independently are selected from separately: methylene dichloride, chloroform, toluene, methyl alcohol, ethanol or THF.
5. method as claimed in claim 2 is characterized in that described method carries out as follows: with shown in organic solvent A 1 dissolution type (II) 3, and 6-dimethyl--1,6-dihydro-1,2; 4,5-tetrazine and basic catalyst obtain containing 3,6-dimethyl--1,6-dihydro-1,2; 4, the solution of 5-tetrazine and basic catalyst places reactor drum with two (trichloromethyl) carbonic ethers with organic solvent A 2 ,-10～0 ℃ of stirring down, slowly drips and describedly contains 3; 6-dimethyl--1,6-dihydro-1,2,4, the solution of 5-tetrazine and basic catalyst; Reinforced finishing, reflux 4～12 hours, TLC follows the tracks of detection, and reaction cools to 4～6 ℃ with reaction solution after finishing; Logical nitrogen drips the solution that the 2-hydroxyanilines is dissolved in organic solvent B again, after dripping off, and reflux 12～48 hours; TLC follows the tracks of detection, and reaction finishes, and after steaming desolventizes, residue recrystallization or column chromatography is obtained the N shown in the formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2; 4; 5-tetrazine-1,4-diformamide compound, described organic solvent A 1 is identical organic solvent with organic solvent A 2; Organic solvent B and organic solvent A 1 are identical or different organic solvent, and it is one of following that described organic solvent A 1, organic solvent B independently are selected from separately: methylene dichloride, chloroform, toluene, methyl alcohol, ethanol or THF; The TV consumption of said organic solvent A 1 and A2 is with shown in the formula (II) 3,6-dimethyl--1, and 6-dihydro-1,2,4,5-tetrazine quality is counted 10～50ml/g; The volumetric usage of said organic solvent B is with shown in the formula (II) 3,6-dimethyl--1, and 6-dihydro-1,2,4,5-tetrazine quality is counted 3～25ml/g.
6. like claim 2 or 5 described methods, it is characterized in that the residue recrystallization described in the described method is to carry out as follows: get the residue that steams after desolventizing and add recrystallization solvent, stirring heating; Refluxed 5～10 minutes, filtered while hot is removed insolubles, the filtrating cooling; Separate out solid; Filter, the filter cake oven dry obtains the N shown in the formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide compound; Said recrystallization solvent is one of following: methyl alcohol, ethanol, sherwood oil, ETHYLE ACETATE, methylene dichloride, chloroform or THF.
7. like claim 2 or 5 described methods, it is characterized in that the residue column chromatography described in the described method carries out as follows: get the residue adding organic solvent C that steams after desolventizing it is dissolved, obtain lysate; In lysate, add the column chromatography silica gel of 1.5～2 times of amounts of residue quality then, behind the mixing, steaming desolventizes; Get the mixture of exsiccant residue and silica gel; Mixture is adorned post, and then with volume ratio 0.5～10: 1 sherwood oil and ethyl acetate mixture are eluent, directly carry out wash-out; TLC follows the tracks of detection; Detect the elutriant that collection contains the compound shown in the formula (I) according to TLC, it is dry to collect liquid, the N shown in the acquisition formula (I)
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1,4-diformamide compound; Said organic solvent C is one of following: methyl alcohol, ethanol, ETHYLE ACETATE, methylene dichloride, chloroform, THF or DMSO 99.8MIN..
8. N as claimed in claim 1
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the application of 4-diformamide (I) in the medicine of preparation prevention or treatment tumor disease is characterized in that said tumour behaviour lung cancer.
9. N as claimed in claim 1
1, N
4-two (2-hydroxy phenyls)-3,6-dimethyl--1,2,4,5-tetrazine-1, the application of 4-diformamide (I) in the medicine of preparation prevention or treatment tumor disease is characterized in that described human lung carcinoma cell line is A-549.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012101003648A CN102702120A (en) | 2012-04-06 | 2012-04-06 | New compound o-hydroxyphenyl tetrazine dicarbonamide, preparation and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012101003648A CN102702120A (en) | 2012-04-06 | 2012-04-06 | New compound o-hydroxyphenyl tetrazine dicarbonamide, preparation and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102702120A true CN102702120A (en) | 2012-10-03 |
Family
ID=46895228
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012101003648A Pending CN102702120A (en) | 2012-04-06 | 2012-04-06 | New compound o-hydroxyphenyl tetrazine dicarbonamide, preparation and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102702120A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103145636A (en) * | 2013-03-25 | 2013-06-12 | 台州职业技术学院 | 1,4-diacyl-3,6-diphenyl-1,4-dihydrotetrazine compound as well as preparation method and application thereof |
| CN105949140A (en) * | 2016-05-06 | 2016-09-21 | 浙江工业大学 | Diphenyl tetrazine dicarboamide compound, preparation and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1223259A (en) * | 1998-09-26 | 1999-07-21 | 杭州民生凯普医药化工有限公司 | Anti-cancer medicine 3,6-dimethyl 1,4-dihydro 1,2,4,5-tetrazine-1,4-dimethyl amides compound and preparation process thereof |
| WO2010042758A2 (en) * | 2008-10-08 | 2010-04-15 | The Scripps Research Institute | Oxazole-pyridazine-oxazole alpha-helix mimetic |
| WO2011137342A1 (en) * | 2010-04-29 | 2011-11-03 | Deciphera Pharmaceuticals, Llc | Cyclopropyl dicarboxamides and analogs exhibiting anti-cancer and anti-proliferative activites |
-
2012
- 2012-04-06 CN CN2012101003648A patent/CN102702120A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1223259A (en) * | 1998-09-26 | 1999-07-21 | 杭州民生凯普医药化工有限公司 | Anti-cancer medicine 3,6-dimethyl 1,4-dihydro 1,2,4,5-tetrazine-1,4-dimethyl amides compound and preparation process thereof |
| WO2010042758A2 (en) * | 2008-10-08 | 2010-04-15 | The Scripps Research Institute | Oxazole-pyridazine-oxazole alpha-helix mimetic |
| WO2011137342A1 (en) * | 2010-04-29 | 2011-11-03 | Deciphera Pharmaceuticals, Llc | Cyclopropyl dicarboxamides and analogs exhibiting anti-cancer and anti-proliferative activites |
Non-Patent Citations (6)
| Title |
|---|
| 《中国医药工业杂志》 20001231 蔡志彬，等 N,N'-二（间二甲胺基苯基）-3,6-二甲基-1,4-二氢-1,2,4,5-四嗪-1,4-二甲酰胺的合成及其抗肿瘤活性 第492页实验部分 1-9 第31卷, 第11期 * |
| WEI-XIAO HU，等: "Synthesis and antitumor activity of s-tetrazine derivatives", 《BIOORGANIC & MEDICINAL CHEMISTRY LETTERS》 * |
| 孙雅泉，等: "1, 6-二氢-S-四嗪催化重排及其反应的研究与氯甲酸三氯甲酯反应", 《合成化学》 * |
| 胡惟孝，等: "1, 6-二氢-S-四嗪与异氰酸酯催化重排反应的研究", 《高等学校化学学报》 * |
| 蔡志彬，等: "N,N’-二（间二甲胺基苯基）-3,6-二甲基-1,4-二氢-1,2,4,5-四嗪-1,4-二甲酰胺的合成及其抗肿瘤活性", 《中国医药工业杂志》 * |
| 饶国武，等，: "Synthesis and structure analysis of N1,N4,3,6-tetramethyl- N1,N4-diphenyl-1,4-dihydro-1,2,4,5-tetrazine-1,4-dicarboxamide", 《JOURNAL OF CHEMICAL RESEARCH》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103145636A (en) * | 2013-03-25 | 2013-06-12 | 台州职业技术学院 | 1,4-diacyl-3,6-diphenyl-1,4-dihydrotetrazine compound as well as preparation method and application thereof |
| CN105949140A (en) * | 2016-05-06 | 2016-09-21 | 浙江工业大学 | Diphenyl tetrazine dicarboamide compound, preparation and application |
| CN105949140B (en) * | 2016-05-06 | 2019-05-31 | 浙江工业大学 | A kind of diphenyl tetrazine diformamide compound and preparation and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102702121B (en) | Hydroxy phenyl tetrazine diformamide and Synthesis and applications between a kind of compound | |
| CN108530343B (en) | Rhein specific group modified organic compound, aryl metal complex thereof, preparation method and application thereof | |
| CN103274987A (en) | 3,3-disubstituted oxoindole derivative, and synthetic method and application thereof | |
| CN104402934A (en) | Preparation method and application of 2-(diphenylphosphineethyl)-(5,6,7,8-tetrahydroquinolinyl)amine ruthenium complexes | |
| CN108570076A (en) | A kind of ruthenium complex and its synthetic method and application containing alkynyl | |
| CN106810560B (en) | A kind of synthetic method of 8- azepine cumarin and its application in anti-tumor drug | |
| CN102702120A (en) | New compound o-hydroxyphenyl tetrazine dicarbonamide, preparation and application thereof | |
| CN105968064B (en) | Two tolyl tetrazine diformamide compounds of one kind and preparation and application | |
| CN102584734B (en) | 3,6-dimethyl-1,2,4,5-tetrazine-1,4-dimethyl amides ramification, preparation method and application | |
| CN102659703A (en) | New compound N<1>,N<4>-di(3,4-dimethoxyphenyl)-3,6-dimethyl-1,2,4,5-tetrazine-1,4-diformamide, preparation thereof and application thereof | |
| CN101007754A (en) | Homotriptycene analog compound and its preparing process | |
| Chou et al. | Steric-influenced fluxionality in monomeric two and three coordinate copper (I) complexes with a trans-chelating bis-pyrazole ligand | |
| CN116253763A (en) | N-heterocyclic carbene metal organic palladium complex of oxidized iso-aporphine alkaloid and synthetic method and application thereof | |
| CN106565755B (en) | Using 1- pyridine -6- methoxy-p-carbolines as copper nitrate (II) chelate and its synthetic method of ligand and application | |
| CN102442964B (en) | N1, N4-di-n-butyl-3, 6-dimethyl-1, 2, 4, 5-tetrazine-1, 4-dicarboximide as well as preparation method and application thereof | |
| CN102558081A (en) | N1, N4-disubstituted phenyl-3, 6-dimethyl-1, 2,4, 5-tetrazine-1, 4-dimethyl amide preparation method | |
| CN111646966B (en) | Fluorescent type ring iridium (III) complex modified by coumarin as well as preparation method and application thereof | |
| CN104098524B (en) | 1-meta-methoxy benzoyl-3-phenyl-Isosorbide-5-Nitrae-dihydro-1,2,4,5-tetrazine and Synthesis and applications | |
| CN113321673A (en) | Preparation method and application of neobynine boric acid compound | |
| CN102268003B (en) | Unsymmetrical poly-substituted porphyrin gold (III) type anticancer compound and preparation method thereof | |
| Sheen et al. | Synthesis of titanium-TADDOLate complexes containing bidentate nitrogen donors and the asymmetric ethylation of benzaldehyde | |
| CN102516313B (en) | Quasi-cis-platinum complex with anticancer activity and two-photon biological development function and preparation method thereof | |
| McGinnis et al. | Synthesis, properties and complexation of (pS)-1-isocyano-2-methylferrocene, the first planar-chiral isocyanide ligand | |
| CN112745356B (en) | Bridged aromatic ruthenium dimer compound and preparation method thereof, aromatic ruthenium compound and preparation method and application thereof | |
| CN105949140B (en) | A kind of diphenyl tetrazine diformamide compound and preparation and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20121003 |