CN106749016A - A kind of ethidium bromide derivative and its preparation and the application in antitumor - Google Patents
A kind of ethidium bromide derivative and its preparation and the application in antitumor Download PDFInfo
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- CN106749016A CN106749016A CN201710011071.5A CN201710011071A CN106749016A CN 106749016 A CN106749016 A CN 106749016A CN 201710011071 A CN201710011071 A CN 201710011071A CN 106749016 A CN106749016 A CN 106749016A
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- C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
- C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
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Abstract
The invention discloses a kind of ethidium bromide derivative and its preparation and the application in antitumor, belong to pharmaceutical technology field.The general structure of ethidium bromide derivative of the present invention is as follows, and R is H or halogen in formula.Ethidium bromide derivative of the invention can realize the targeting enrichment to tumour cell, and possess Mitochondrially targeted effect, have the effect of good Selective depression to tumour cell, can be used to prepare antineoplastic.Ethidium bromide derivative of the invention is prepared simply efficiently, with great medical applications value.
Description
Technical field
The invention belongs to pharmaceutical technology field, and in particular to a kind of ethidium bromide derivative and its prepare with antitumor
Application.
Background technology
Tumour is the major disease for threatening human health, and neoplasm targeted therapy is clinical medicine, biological medicine and life section
Learn the key subjects of research field.Drug therapy is to realize Tumor growth inhibition and improve patients ' life quality have an efficacious prescriptions
Method, is widely used in clinical therapy of tumor.Traditional antineoplastic includes adriamycin, taxol, cis-platinum, camptothecine
Deng.But, these antineoplastics are poor to the selectivity of oncotherapy, so as to cause the secondary work of larger poison over the course for the treatment of
With.Also, these medicines can not effectively reach action target spot, cause relatively low therapeutic effect.In order to improve to tumour
Therapeutic effect, be usually taken improve dosage method, and further result in tumour cell produce MDR, ultimately result in
The failure of oncotherapy.Therefore, antineoplastic is improved to tumour cell and the ability to the targeting enrichment of its action site, is opened
Efficient antineoplastic is sent by the process of greatly propulsion oncotherapy.
The content of the invention
It is an object of the invention to provide a kind of ethidium bromide derivative and preparation method thereof and preparing antineoplastic
Application in thing.
The purpose of the present invention is achieved through the following technical solutions：
A kind of ethidium bromide derivative, its general structure is as follows：
Wherein, R is H or halogen, and described halogen is F, Cl, Br.Preferably, R is H or Cl.
The preparation method of described ethidium bromide derivative, comprises the following steps：
(1) ethidium bromide (EB) is dissolved in volume ratio for 0.005-0.05:0.5-2:Trifluoroacetic acid/acetonitrile/the dichloro of 3-5
In methane blended solvent, lead to nitrogen protection, stirred 5-10 minutes under the conditions of 0-4 DEG C.
(2) natrium nitrosum is added in step (1) gained mixture, is kept for 0 DEG C -4 DEG C react 5-10 minutes.
(3) sulfamic acid is added in step (2) gained reaction solution, is kept for 0-4 DEG C react 5-10 minutes.
(4) by N, N- diethylanilines or double (2- chloroethyls) aniline of N, N- are dissolved in volume ratio for 0.005-0.05:
0.5-2:In the trifluoroacetic acid/acetonitrile/dichloromethane mixed solvent of 3-5, it is added in step (3) gained reaction solution, keeps 0-4
DEG C reaction obtains ethidium bromide derivative in 60-120 minutes.
Trifluoroacetic acid in above-mentioned trifluoroacetic acid/acetonitrile/dichloromethane mixed solvent, acetonitrile, dichloromethane volume ratio it is excellent
Elect 0.01 as:1:4.
A kind of above-mentioned ethidium bromide derivative pharmaceutically acceptable salt.Described ethidium bromide derivative is to tumour cell
And its subcellular organelle has good targeting, and can effectively suppress the growth of tumour cell.Based on this, described bromination
Second ingot derivative or its pharmaceutically acceptable salt can be used to prepare antineoplastic.
A kind of antineoplastic, comprising above-mentioned ethidium bromide derivative or its pharmaceutically acceptable salt, also comprising described
Ethidium bromide derivative pharmaceutically acceptable carrier or excipient.
The present invention has the following advantages that and effect relative to prior art：Ethidium bromide derivative of the invention can be realized
Targeting enrichment to tumour cell, and possess Mitochondrially targeted effect, there is good Selective depression to tumour cell
Effect, by changing mitochondrial membrane potential, improve intracellular oxyradical level, reduce intracellular reproducibility paddy
The sweet peptide level of Guang, reduces intracellular ATP levels, causes the release of Intramitochondrial cromoci, and final inducing cell withers
Die.In addition, this quasi-molecule is prepared simply efficiently, with great medical applications value.
Brief description of the drawings
Fig. 1 is the electrospray ionization mass spectrum figure of compound 1.
Fig. 2 is the electrospray ionization mass spectrum figure of compound 2.
Fig. 3 be breast cancer cell and African green monkey kidney cell with after the medium culture containing antitumoral compounds 2 with
The fluorescence microscopy figure of Hoechst33342 marks.
Fig. 4 is marked with MitoTracker Green after breast cancer cell uses the medium culture containing antitumoral compounds 2
Fluorescence microscopy figure.
Fig. 5 is EB, antitumoral compounds 1 and antitumoral compounds 2 respectively to (A) breast cancer cell and (B) cercopithecus aethiops
The cytoactive of nephrocyte suppresses figure.
Fig. 6 be breast cancer cell with after the medium culture different time containing antitumoral compounds 2 with JC-1 dye markers
Fluorescence microscopy figure.1-4 is respectively from left to right in figure：1 is burnt JC-1 monomer fluorescences copolymerization, and 2 is common for JC-1 polymer fluorescents
Focus on, 3 for 1 and 2 overlap, 4 is the fluorescence intensity of the JC-1 at arrow in corresponding 3.
Fig. 7 be breast cancer cell with after the medium culture different time containing antitumoral compounds 2 with JC-1 dye markers
Fluorescence statistical analysis figure.Figure A is breast cancer cell through antitumoral compounds 1 or the treatment different time of antitumoral compounds 2
The Fluorescence Ratio of intracellular JC-1 monomers and JC-1 polymer afterwards, figure B is breast cancer cell respectively through EB, antitumoral compounds 1
Or antitumoral compounds 2 process 6 hours after intracellular JC-1 monomers fluorescence intensity.
Fig. 8 is breast cancer cell with after the medium culture containing EB, antitumoral compounds 1 and antitumoral compounds 2 respectively
With the fluorescence microscopy figure that DCFH-DA dyestuffs are detected.
Fig. 9 is breast cancer cell with after the medium culture containing EB, antitumoral compounds 1 and antitumoral compounds 2 respectively
With the fluorescence statistical analysis figure that DCFH-DA dyestuffs are detected.
Figure 10 is breast cancer cell with after the medium culture containing EB, antitumoral compounds 1 and antitumoral compounds 2 respectively
The content analysis figure of intracellular reductive glutathione.
Figure 11 is breast cancer cell with after the medium culture containing EB, antitumoral compounds 1 and antitumoral compounds 2 respectively
Intracellular ATP content analysis figures.
Figure 12 is cromoci of the breast cancer cell in born of the same parents' cytoplasm after the medium culture containing antitumoral compounds 1
Ratio analysis figure.
Figure 13 is cromoci phase of the breast cancer cell in mitochondria after the medium culture containing antitumoral compounds 1
To content analysis figure.
Figure 14 is breast cancer cell with after the medium culture containing EB, antitumoral compounds 1 and antitumoral compounds 2 respectively
The ratio analysis figure of intracellular apoptosis enzyme -3.
Figure 15 is that mouse is cultivated 13 days after making knurl mouse with the treatment of PBS, EB, antitumoral compounds 1 or antitumoral compounds 2
Interior gross tumor volume changes with time figure.
Figure 16 is that mouse is cultivated 13 days after making knurl mouse with the treatment of PBS, EB, antitumoral compounds 1 or antitumoral compounds 2
Tumour figure afterwards.
Figure 17 is that mouse is cultivated 13 days after making knurl mouse with the treatment of PBS, EB, antitumoral compounds 1 or antitumoral compounds 2
Tumor weight figure afterwards.
Figure 18 is that mouse is cultivated 13 days after making knurl mouse with the treatment of PBS, EB, antitumoral compounds 1 or antitumoral compounds 2
Interior Mouse Weight changes with time figure.
Specific embodiment
Further detailed description, but embodiments of the present invention not limited to this are done to the present invention with reference to embodiment.
Embodiment 1
The synthesis of antineoplastic compounds 1：
(1) it is 0.01 that EB (230mg) is dissolved in 50mL volume ratios:1:4 trifluoroacetic acid/acetonitrile/dichloromethane mixing is molten
In agent, the logical nitrogen protection of the mixture is stirred 10 minutes under the conditions of 0 DEG C.
(2) natrium nitrosum (150mg) is added in step (1) gained mixture, stirring reaction 5 under the conditions of being kept for 0 DEG C
Minute.
(3) sulfamic acid (200mg) is added in step (2) gained reaction solution, stirring reaction 10 under the conditions of being kept for 0 DEG C
Minute.
(4) by N, it is 0.01 that N- diethylanilines (200 μ L) are dissolved in 2mL volume ratios:1:4 trifluoroacetic acid/acetonitrile/bis-
In chloromethanes mixed solvent, it is added in step (3) gained reaction solution, 0 DEG C of holding stirring reaction 60 minutes.
(5) after reaction terminates, step (4) gained reaction solution dchloromethane is washed with water three times, organic phase is received
Collection gets up, and is dried overnight with dichloromethane.
(6) product that will be obtained is purified with column chromatography method, and eluent is that volume ratio is 1:9 ethanol/methylene is mixed
Bonding solvent.
(7) electrospray ionization mass spectrum characterization adduct molecule structure is used, and the product for obtaining is protected under the conditions of -20 DEG C of lucifuges
Deposit.
Electrospray ionization mass spectrum result is as shown in Figure 1, it was demonstrated that the successful synthesis of compound 1.
Embodiment 2
The synthesis of antineoplastic compounds 2：
(1) it is 0.01 that EB (230mg) is dissolved in 50mL volume ratios:1:4 trifluoroacetic acid/acetonitrile/dichloromethane mixing is molten
In agent, the logical nitrogen protection of the mixture is stirred 5 minutes under the conditions of 0 DEG C.
(2) natrium nitrosum (150mg) is added in step (1) gained mixture, 0 DEG C of holding stirring reaction 10 minutes.
(3) sulfamic acid (200mg) is added in step (2) gained reaction solution, 0 DEG C of holding stirring reaction 5 minutes.
(4) by N, it is 0.01 that double (2- chloroethyls) aniline (200 μ L) of N- are dissolved in 2mL volume ratios:1:4 trifluoroacetic acid/
In acetonitrile/dichloromethane mixed solvent, it is added in step (3) gained reaction solution, 0 DEG C of holding stirring reaction 60 minutes.
(5) after reaction terminates, step (4) gained reaction solution dchloromethane is washed with water three times, organic phase is received
Collection gets up, and is dried overnight with dichloromethane.
(6) product that will be obtained is purified with column chromatography method, and eluent is that volume ratio is 1:9 ethanol/methylene is mixed
Bonding solvent.
(7) electrospray ionization mass spectrum characterization adduct molecule structure is used, and the product for obtaining is protected under the conditions of -20 DEG C of lucifuges
Deposit.
Electrospray ionization mass spectrum result is as shown in Figure 2, it was demonstrated that the successful synthesis of compound 2.
Embodiment 3
By breast cancer cell (4T1 cells) and African green monkey kidney cell (COS7 cells) respectively with 1 × 105Individual cells/well
Density be inoculated into copolymerization Jiao capsule in, under the conditions of 37 DEG C in 1mL RPMI-1640 culture mediums cultivate.After 24 hours, will be anti-
Neoplastic compound 2 is dissolved in the medium, and antitumor chemical combination is contained to 1mL is added in breast cancer cell and African green monkey kidney cell
The culture medium of thing 2 (20 μm of ol/L).After culture 2 hours, the culture medium containing antitumoral compounds 2 is suctioned out, PBS cushioning liquid
Cell washing is marked into nucleus three times with Hoechst 33342 afterwards, it is then intracellular anti-with confocal laser scanning microscope
The fluorescence intensity of neoplastic compound 2.
Result is as shown in figure 3, relative to African green monkey kidney cell, antitumoral compounds 2 have more preferable to breast cancer cell
Enrichment, illustrate that antitumoral compounds 2 have good targeting enrichment to tumour cell.
Embodiment 4
By breast cancer cell with 1 × 105In density inoculation copolymerization Jiao's capsule of individual cells/well, trained in 1mL under the conditions of 37 DEG C
Support culture in base.After 24 hours, antitumoral compounds 2 are dissolved in the medium, it is anti-to adding 1mL to contain in breast cancer cell
The culture medium of neoplastic compound 2 (20 μm of ol/L).After culture 2 hours, the culture medium containing antitumoral compounds 2 is suctioned out, PBS
Cell is washed three times and uses mitochondrial dye (MitoTracker Green) labeled mitochondria afterwards by cushioning liquid, then uses laser
The distribution of the intracellular antitumoral compounds 2 of confocal microscopy.
Result as shown in figure 4, the fluorescence of antitumoral compounds 2 overlaps well with mitochondrial dye fluorescence in tumour cell,
Prove that antitumoral compounds 2 have the Mitochondrially targeted effect of tumour cell.
Embodiment 5
Breast cancer cell and African green monkey kidney cell are seeded in 96 orifice plates with the density of 6000 cells/wells respectively, are used
100 μ L medium cultures 24 hours.Then, the various concentrations that will be prepared with culture medium respectively containing EB, antitumoral compounds 2,
The μ L of solution 100 of antitumoral compounds 1 are added separately in each hole.All cells are cultivated 48 hours for 37 DEG C under the conditions of lucifuge.
The MTT (MTT is dissolved in PBS) of 20 μ L 5mg/mL is then added in each hole.After co-culturing 4 hours, training is suctioned out
Base is supported, 150 μ L dimethyl sulfoxides are added.ELIASA measures the light absorption value of 570 nanometers in each hole, calculates cell survival rate, enters
And EB, antitumoral compounds 2, antitumoral compounds 1 are obtained to breast cancer cell and the toxicity of African green monkey kidney cell.
Result is as shown in figure 5, antitumoral compounds 2, antitumoral compounds 1 are to breast cancer cell and African green monkey kidney cell
With great toxicity.And relative to African green monkey kidney cell, antitumoral compounds 2, antitumoral compounds 1 are thin to breast cancer
Born of the same parents have bigger toxicity；Antitumoral compounds 1 possess bigger cytotoxicity relative to antitumoral compounds 2.So as to prove
Antitumoral compounds 2 and antitumoral compounds 1 have toxicity higher to tumour cell.
Embodiment 6
By breast cancer cell with 1 × 105The density of individual cells/well is seeded in copolymerization Jiao's capsule, in 1mL under the conditions of 37 DEG C
Cultivated in culture medium.After 24 hours, by antitumoral compounds (2 or 1) dissolving in the medium, 1mL is added to breast cancer cell
Culture medium containing antitumoral compounds (20 μm of ol/L).After culture 1 hour, the culture medium containing antitumoral compounds is inhaled
Go out, PBS cushioning liquid washs cell three times, cell is further cultured for respectively 1 hour, 2 hours, 6 hours, 12 hours and 23 hours
Afterwards, half an hour is dyeed with mitochondrial dye (JC-1), then with the intracellular JC-1 fluorescence of confocal laser scanning microscope
Distribution.At the same time, intracellular JC-1 fluorescence intensities are analyzed with ImageJ to change with time.
Result as shown in fig. 6, when antitumoral compounds 2 and cytosis short period, tumour cell mitochondrial membrane potential
It is not changed in, and the destruction for increasing the mitochondrial membrane of tumour cell with incubation time strengthens.At the same time, as shown in Figure 7 A, resist
Neoplastic compound 1 has stronger destruction to cancer cell mitochondrial membrane compared with antitumoral compounds 2.Therefore, antitumoral compounds 2
With the effect that antitumoral compounds 1 all possess destruction mitochondrial membrane potential.
Embodiment 7
By breast cancer cell with 1 × 105The density of individual cells/well is seeded in six orifice plates, is cultivated in 1mL under the conditions of 37 DEG C
Cultivated in base.After 24 hours, EB, antitumoral compounds 2 and antitumoral compounds 1 are dissolved in the medium respectively, to mammary gland
1mL is separately added into cancer cell containing EB, the culture medium of antitumoral compounds 2 or antitumoral compounds 1 (20 μm of ol/L) or only
Blank cultures are added as blank.After culture 1 hour, EB, antitumoral compounds 2 and antitumoral compounds 1 will be contained
Culture medium, blank cultures suction out, PBS cushioning liquid by cell wash three times, after cell is further cultured for into 6 hours respectively, use
Mitochondrial dye (JC-1) dyes half an hour, then with the distribution of the intracellular JC-1 monomer fluorescences of flow cytometry analysis.
As shown in Figure 7 B, antitumoral compounds 2 and antitumoral compounds 1 have a JC-1 monomer fluorescences higher to result, and EB
It is almost identical with blank control group, so as to demonstrate again that antitumoral compounds 2 and antitumoral compounds 1 have mitochondrial membrane potential
The ability of destruction.
Embodiment 8
By breast cancer cell with 1 × 105The density of individual cells/well is seeded in copolymerization Jiao's capsule, in 1mL under the conditions of 37 DEG C
Cultivated in culture medium.After 24 hours, respectively by EB, antitumoral compounds 2 and antitumoral compounds 1 dissolve in the medium, to
Be separately added into breast cancer cell 1mL containing EB, the culture medium of antitumoral compounds 2 or antitumoral compounds 1 (20 μm of ol/L) or
Person only adds blank cultures as blank.After culture 1 hour, EB, antitumoral compounds 2 and antitumor chemical combination will be contained
The culture medium of thing 1, blank cultures are suctioned out, and PBS cushioning liquid washs cell three times, after cell is further cultured for into 6 hours respectively
Half an hour is co-cultured with living radical probe DCFH-DA, then the intracellular fluorescence intensity of confocal laser scanning microscope.
At the same time, with intensity of cellular fluorescence under ImageJ analysis different conditions.
Result as shown in figure 8, blanc cell and with EB co-culture after cell in fluorescence intensity it is relatively low, and with it is antitumor
The fluorescence intensity in cell after compound 2 and the co-cultivation of antitumoral compounds 1 is higher, so as to prove the He of antitumoral compounds 2
Antitumoral compounds 1 have the ability of enhancing intracellular reactive free radical.At the same time, as shown in figure 9, and antitumoral compounds
1 effect after intracellular living radical be higher than and antitumoral compounds 2 act on after living radical, it was demonstrated that with it is antitumor
Compound 1 possesses stronger antitumor action.
Embodiment 9
By breast cancer cell with 1 × 105The density of individual cells/well is seeded in six orifice plates, is cultivated in 1mL under the conditions of 37 DEG C
Cultivated in base.After 24 hours, EB, antitumoral compounds 2 and antitumoral compounds 1 are dissolved in the medium respectively, to mammary gland
1mL is added containing EB, the culture medium of antitumoral compounds 2 or antitumoral compounds 1 (20 μm of ol/L) in cancer cell or is only added
Blank cultures are used as blank.After culture 1 hour, by the training containing EB, antitumoral compounds 2 and antitumoral compounds 1
Support base, blank cultures to suction out, PBS cushioning liquid washs cell three times, by cell after cell is further cultured for respectively 24 hours
Cracking, after 6000 revs/min are centrifuged 5 minutes, take the μ L of supernatant 50 and is added in 200 μ L Ellman detection reagents, uses ELIASA
Detect the absorbance in 406 nanometers after different sample treatments.
Result is as shown in Figure 10, going back in the tumor cell after antitumoral compounds 2 and the effect of antitumoral compounds 1
Originality glutathione level is substantially reduced, and demonstrates again that antitumoral compounds 2 and antitumoral compounds 1 possess the intracellular oxygen of regulation
Change the ability of reduction internal pressure.
Embodiment 10
By breast cancer cell with 1 × 105The density of individual cells/well is seeded in six orifice plates, is cultivated in 1mL under the conditions of 37 DEG C
Cultivated in base.After 24 hours, EB, antitumoral compounds 2 and antitumoral compounds 1 are dissolved in the medium respectively, to mammary gland
1mL is added containing EB, the culture medium of antitumoral compounds 2 or antitumoral compounds 1 (20 μm of ol/L) in cancer cell or is only added
Blank cultures are used as blank.After culture 1 hour, EB, the training of antitumoral compounds 2 and antitumoral compounds 1 will be contained
Support base, blank cultures to suction out, PBS cushioning liquid washs cell three times, by cell after cell is further cultured for respectively 24 hours
Cracking, the ATP contents in different samples are detected with ATP detection kits.
Result is as shown in figure 11, in the tumor cell after antitumoral compounds 2 and the effect of antitumoral compounds 1
ATP levels are substantially reduced, it was demonstrated that antitumoral compounds 2 and antitumoral compounds 1 possess the energy for suppressing ATP synthesis in tumour cell
Power.
Embodiment 11
By breast cancer cell with 1 × 105The density of individual cells/well is seeded in six orifice plates, is cultivated in 1mL under the conditions of 37 DEG C
Cultivated in base.After 24 hours, antitumoral compounds 1 are dissolved in the medium, contain antitumor to 1mL is added in breast cancer cell
The culture medium of compound 1 (20 μm of ol/L) only adds blank cultures as blank.After culture 6 hours, will contain
The culture medium of antitumoral compounds 1, blank cultures are suctioned out, and PBS cushioning liquid washs cell three times, and cell is trained again respectively
Albumen is extracted after supporting 24 hours, the relative of cromoci contains in western blot analysis tumor cell matter and mitochondria
Amount.
Result is as shown in figure 12, the cromoci level in tumor cell matter after being acted on antitumoral compounds 1
Higher than control blanc cell.At the same time, as shown in figure 13, in the tumour cell mitochondria after being acted on antitumoral compounds 1
Cromoci level less than control blanc cell.These results suggest that, antitumoral compounds can with induced cytochrome C from
Discharged in mitochondria.
Embodiment 12
By breast cancer cell with 1 × 105The density of individual cells/well is seeded in six orifice plates, is cultivated in 1mL under the conditions of 37 DEG C
Cultivated in base.After 24 hours, EB, antitumoral compounds 2 and antitumoral compounds 1 are dissolved in the medium respectively, to mammary gland
1mL is added containing EB, the culture medium of antitumoral compounds 2 or antitumoral compounds 1 (20 μm of ol/L) in cancer cell or is only added
Blank cultures are used as blank.After culture 6 hours, by the training containing EB, antitumoral compounds 2 and antitumoral compounds 1
Support base, blank cultures to suction out, PBS cushioning liquid washs cell three times, and egg is extracted after cell is further cultured for into 24 hours respectively
In vain, the relative amount of apoptosis enzyme -3 is activated in western blot analyses tumour cell.
Result as shown in figure 14, is activated in the tumour cell after being acted on antitumoral compounds 2 and antitumoral compounds 1 and withered
Die enzyme -3 expression improve, it was demonstrated that antitumoral compounds 2 and antitumoral compounds 1 can be with induced tumor programmed cell apoptosis.
Embodiment 13
5~6 weeks small white mouse (BALB/c) the μ L of big leg outer side hypodermic injection 100 containing number of cells be 1 × 106Breast cancer
Cell PBS suspensions make knurl, when gross tumor volume grows to about 200mm3When.Mouse is randomly divided into 4 groups.PBS and EB, antitumor chemical combination
Thing 2 and antitumoral compounds 1.Respectively in the material that injection in first day and the 5th day is relative.Gross tumor volume is every other day measured, is claimed
Amount Mouse Weight change.Gross tumor volume is according to V=W2× L/2 formula are calculated, and wherein W is shorter tumor width, and L is more long
Tumor width, by relative tumour volume (V/V0, V is real-time gross tumor volume, V0It is treatment pre-neoplastic volume) reflect tumour body
Long-pending situation of change.After mouse was cultivated by the 13rd day, mouse tumor is peeled off, mouse tumor weight is measured respectively.
Result is shown in Figure 15, Figure 16 and Figure 17, the mouse tumor after being treated through antitumoral compounds 2 and antitumoral compounds 1
Growth is suppressed, and antitumoral compounds 1 have more superior tumor inhibition effect relative to neoplastic compound 2.It is same with this
When, as shown in figure 18, mice weights do not have in a substantial change with therapeutic effect, illustrate antitumoral compounds 2 and antitumoral compounds 1
There is no strong toxic and side effect to mouse.
Above-described embodiment is the present invention preferably implementation method, but embodiments of the present invention are not by above-described embodiment
Limitation, it is other it is any without departing from Spirit Essence of the invention and the change, modification, replacement made under principle, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (8)
1. a kind of ethidium bromide derivative, it is characterised in that：General structure is as follows：
Wherein, R is H or halogen.
2. ethidium bromide derivative according to claim 1, it is characterised in that：R is H or Cl.
3. the preparation method of the ethidium bromide derivative described in claim 1, it is characterised in that：Comprise the following steps：
(1) ethidium bromide is dissolved in volume ratio for 0.005-0.05:0.5-2:Trifluoroacetic acid/the acetonitrile of 3-5/dichloromethane mixing
In solvent, lead to nitrogen protection, stirred 5-10 minutes under the conditions of 0-4 DEG C；
(2) natrium nitrosum is added in step (1) gained mixture, is kept for 0-4 DEG C react 5-10 minutes；
(3) sulfamic acid is added in step (2) gained reaction solution, is kept for 0-4 DEG C react 5-10 minutes；
(4) by N, N- diethylanilines or double (2- chloroethyls) aniline of N, N- are dissolved in volume ratio for 0.005-0.05:0.5-
2:In the trifluoroacetic acid/acetonitrile/dichloromethane mixed solvent of 3-5, it is added in step (3) gained reaction solution, is kept for 0-4 DEG C instead
Answer 60-120 minutes and obtain ethidium bromide derivative.
4. the preparation method of ethidium bromide derivative according to claim 3, it is characterised in that：Described trifluoroacetic acid/
Trifluoroacetic acid, acetonitrile, the volume ratio of dichloromethane are 0.01 in acetonitrile/dichloromethane mixed solvent:1:4.
5. the ethidium bromide derivative pharmaceutically acceptable salt described in a kind of claim 1.
6. the salt described in the ethidium bromide derivative or claim 5 described in claim 1 in antineoplastic is prepared should
With.
7. a kind of antineoplastic, it is characterised in that：Comprising ethidium bromide derivative or claim 5 described in claim 1
Described salt.
8. antineoplastic according to claim 7, it is characterised in that：Spread out comprising the ethidium bromide described in claim 1
Acceptable carrier or excipient on biopharmacy.
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| CN108395460A (en) * | 2018-01-31 | 2018-08-14 | 广州医科大学 | A kind of weary oxygen activation adriamycin prodrug and preparation method thereof |
| CN108395460B (en) * | 2018-01-31 | 2020-05-26 | 广州医科大学 | Hypoxia activated adriamycin prodrug and preparation method thereof |
| WO2023207982A1 (en) * | 2022-04-29 | 2023-11-02 | 上海交通大学医学院附属瑞金医院 | Pharmaceutical composition of ethidium bromide, and use thereof in treating cancer |
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