CN104606207A - Application of sulfamethoxazole in preparing medicament for inhibiting tumor cell metastasis and diffusion - Google Patents
Application of sulfamethoxazole in preparing medicament for inhibiting tumor cell metastasis and diffusion Download PDFInfo
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- CN104606207A CN104606207A CN201510068148.3A CN201510068148A CN104606207A CN 104606207 A CN104606207 A CN 104606207A CN 201510068148 A CN201510068148 A CN 201510068148A CN 104606207 A CN104606207 A CN 104606207A
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- 229960005404 sulfamethoxazole Drugs 0.000 title claims abstract description 41
- JLKIGFTWXXRPMT-UHFFFAOYSA-N sulphamethoxazole Chemical compound O1C(C)=CC(NS(=O)(=O)C=2C=CC(N)=CC=2)=N1 JLKIGFTWXXRPMT-UHFFFAOYSA-N 0.000 title claims abstract description 40
- 239000003814 drug Substances 0.000 title claims abstract description 27
- 210000004881 tumor cell Anatomy 0.000 title claims abstract description 20
- 206010027476 Metastases Diseases 0.000 title claims abstract description 10
- 230000009401 metastasis Effects 0.000 title claims abstract description 10
- 230000002401 inhibitory effect Effects 0.000 title abstract description 5
- 238000009792 diffusion process Methods 0.000 title abstract description 4
- 210000004027 cell Anatomy 0.000 claims abstract description 73
- 206010009944 Colon cancer Diseases 0.000 claims description 22
- 208000029742 colonic neoplasm Diseases 0.000 claims description 21
- 230000005764 inhibitory process Effects 0.000 claims description 6
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- 238000002360 preparation method Methods 0.000 description 3
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- 230000001629 suppression Effects 0.000 description 3
- 102000000905 Cadherin Human genes 0.000 description 2
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- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 102000016359 Fibronectins Human genes 0.000 description 2
- 108010067306 Fibronectins Proteins 0.000 description 2
- KYRVNWMVYQXFEU-UHFFFAOYSA-N Nocodazole Chemical compound C1=C2NC(NC(=O)OC)=NC2=CC=C1C(=O)C1=CC=CS1 KYRVNWMVYQXFEU-UHFFFAOYSA-N 0.000 description 2
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- 239000002609 medium Substances 0.000 description 2
- 229950006344 nocodazole Drugs 0.000 description 2
- -1 sulfamethoxazole compound Chemical class 0.000 description 2
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses application of sulfamethoxazole in preparing medicament for inhibiting tumor cell metastasis and diffusion. According to the application disclosed by the invention, the medicament prepared by utilizing the sulfamethoxazole is capable of inhibiting the metastasis ability of tumor cells; as being a low toxic medicament, the sulfamethoxazole can have large effect of blocking the process of tumor diffusion; meanwhile, other relative tumor medicaments are also adopted to carry out treatment, and the mortality of normal cell can be greatly reduced; while the traditional tumor medicaments are mostly cytotoxic medicaments, a great amount of the normal cells die together while the tumor cells are killed.
Description
Technical field
The present invention relates to sulfamethoxazole in the new purposes of pharmaceutical field, be specifically related to the application of sulfamethoxazole in the medicine preparing inhibition tumor cell metastasis and extension.
Background technology
Sulfamethoxazole, sulfonamides is class antibacterials of synthetic.Cure mainly escherichia coli and microbial acute, the Chronic Urinary Tract Infection of deformed rod; The prevention of epidemic cerebrospinal meningitis caused by meningococcus; Otitis media etc. caused by hemophilus influenza.Sulfonamides all has inhibitory action to many gram positive bacterias and some gram negative bacterias, Nocardia, chlamydiaceae and some protozoon (as plasmodium and ameba).
Other pharmacological actions of sulfamethoxazole are not yet found in existing open source literature.
Summary of the invention
Goal of the invention: the object of the invention is to provide the application of a kind of sulfamethoxazole in the medicine preparing inhibition tumor cell metastasis and extension.
Technical scheme: the application of sulfamethoxazole in the medicine preparing inhibition tumor cell metastasis and extension.
Preferably, described tumor cell is colon cancer cell.
Preferably, wherein the concentration of sulfamethoxazole is 50 ~ 60uM..
Inventor be experimental studies have found that by cell aspect, and the transfer ability of sulfamethoxazole to tumor cell has certain inhibitory action.Current is oral for the general instructions of taking of existing indication: be divided into fugitive, middle effect and long-acting class according to the length of drug treating time.Oral sulphonamides is mainly at intestinal absorption, and blood drug level peaked in 4 ~ 6 hours.
It is reference frame that the present invention needs according to existing preparation instructions of taking, and existing cell aspect experiment basis determines new clinical administration and dosage according to new indication curative effect and the design of follow-up related experiment.
The present invention by after tumor cell collection is cultivated, recycling medicine irritation cell, observation of cell form the area of quantitative analysis cell layer, namely cell can with the area of other cells contacting, thus judge the diffusivity of tumor cell.Cellular layer area is more conducive to being formed with other cells being connected more greatly.Cell migration needs the cooperation of internal and external factor.Outside factor refers to extracellular signaling molecule.Internal factor is the signal transducting system of phalangeal cell and the cytoskeleton of execution motion and molecular motor then, also has the various molecules participating in talin formation.Extracellular signal needs endocellular signal molecule relay after completing its mission in conjunction with after birth receptor, movable information is passed to further the executable unit of cell migration---cytoskeleton and molecular motor.Miscellaneous endocellular signal molecule can interact, and affects the distribution of these two kinds of molecules aftermentioned, structure and activity, reaches the object of intense adjustment cell movement.As can be seen here, the transfer of tumor cell, and the close relation between cell itself and cell.
Cancerous cell is compared with its homology normal structure, and intercellular cohesiveness reduces, therefore cancerous cell easily disperses in vivo and shifts.Normal cell outer by fibronectin be a kind of extracellular adhesive glycoprotein, it enhance the adhesion between cell and extracellular matrix.The fibronectin of cancerous cell significantly reduces or lacks, and obstacle occurs in cadherin synthesis, thus destroys the adhesion between cell and substrate and between cell and cell, and therefore cancerous cell has the attribute being easy to invade profit tissue and transfer.The increase contact area enhanced between each cell of the area of cancerous cell is more conducive to adhesion each other and attachment.
Cause although the transfer of tumor cell is formed jointly by many factors, but with the area of cellular layer, namely cell can with the foundation of the area of other cells contacting as cell transfer ability size, in this, as the method for measuring of Cell tracking intensity, there is the advantage that with low cost, easy and simple to handle, result is easily sentenced, the cell area that it shows increases the most directly observation and distinguishes, after mainly being stimulated by dosing exactly, immunostaining is carried out to Cell tracking molecule E-cadherin protein, then carry out quantitative assay.Cellular layer area is more conducive to being formed with other cells being connected more greatly.
The present invention thinks that sulfamethoxazole is experimental subject, and its merit rating shifted at inhibition tumor cell is considered in assessment.Main Basis is changed to specifically with cell area before and after medicine irritation.
Beneficial effect: 1, the transfer ability of the present invention's drug on tumor cell of utilizing sulfamethoxazole to prepare suppresses, sulfamethoxazole is low cytotoxic drug, larger blocking effect can be played in tumor diffusion process, meanwhile adopt other related neoplasms medicines to treat again, Normocellular mortality can be reduced in a large number; And existing tumour medicine mostly is cell toxicity medicament, while killing tumor cell, a large amount of normal cell is also dead together; 2, sulfamethoxazole used in the present invention is into the existing medical compounds of Drug Storage, and the R&D cycle can obviously reduce compared with the cycle of new drug development.
Accompanying drawing explanation
Fig. 1 is the colon cancer cell figure after the process of 10uM concentration sulfamethoxazole;
Fig. 2 is the colon cancer cell figure after the process of 20uM concentration sulfamethoxazole;
Fig. 3 is the colon cancer cell figure after the process of 50uM concentration sulfamethoxazole;
Fig. 4 is the colon cancer cell figure after the process of 60uM concentration sulfamethoxazole;
Fig. 5 is the colon cancer cell figure after the process of 80uM concentration sulfamethoxazole;
Fig. 6 is the colon cancer cell figure after the process of 100uM concentration sulfamethoxazole;
Fig. 7 is positive controls, by the colon cancer cell figure after 10uM concentration nocodazole process;
Fig. 8 is negative control group.
Detailed description of the invention
Below by accompanying drawing, technical solution of the present invention is described in detail, but protection scope of the present invention is not limited to described embodiment.
Embodiment 1: sulfamethoxazole suppresses the application in the medicine of colon cancer cell metastasis and extension in preparation.Specific experiment scheme is as follows:
1, experiment material
That the experimental stage of the present invention adopts is colon cancer cell (HT29), and the medicine that positive control adopts is nocodazole (experimental concentration is 10uM).Negative control group is not through any process.
Wherein experiment adopts sulfamethoxazole Compound C
10h
11n
3o
3s, CAS NO. is 723-46-6, and concentration is respectively 100uM, 80uM, 60uM, 50uM, 20uM, 10uM.
2, experimental technique
To the cultivation colon cancer cell of 24 hours (cell density: 10
5) carry out medicine irritation process in 2 hours:
(1) by centrifugal for good for state in culture dish colon cancer cell, add appropriate culture fluid and make cell suspending liquid;
(2) obtained cell suspension counting, according to 10
5density add in 24 porose discs, put into the CO of 37 DEG C
2cultivate in incubator;
After (3) 24 hours, change liquid, the sulfamethoxazole compound adding respective amount in culture fluid is configured to experiment desired concn, and positive control medicine, then continues cultivation 2 hours;
(4) cultivate after 2 hours and the PFA solution lower 37 DEG C of cell in 2% is fixed 10 minutes, then carry out fluorescence staining.
3, experimental result process
Observe the colon cancer cell after fluorescence staining, the picture Image J process of observation is calculated the data of its cell area before and after dosing, and Ratio is the ratio of experimental group and negative control group.The results are shown in Table 1.
Table 1 sulfamethoxazole is to the size of HT29 cytositimulation after 2 hours
Prove through experimental data, sulfamethoxazole makes the obvious scaling up of colon cancer cell surface area under concentration 60uM, and wherein effect is 50 ~ 60uM preferably, proves that sulfamethoxazole can suppress the application in the medicine of colon cancer cell metastasis and extension in preparation.
Embodiment 2:Transwell Cell migration assay measures sulfamethoxazole to be affected colon cancer cell transfer ability.
By 1 × 10
5hT29 cell be seeded in Transwell cell, bottom adopts 12.0 μm of films, continue cultivate.After cell attachment, upper strata cell adds the DMEM complete medium of variable concentrations sulfamethoxazole, for maintaining osmotic pressure, need add 0.05%-0.2%BSA, and bottom chamber adds the DMEM complete medium containing 15%FBS and continues to cultivate 24h.Carefully wipe with cotton swab the cell that migration does not occur film upper surface, the cell migrating to film lower surface dyes with Giemsa after 2%PFA fixes 10 minutes at 37 DEG C.Random selecting 10 visuals field under high power microscope, counting migrating cell.
Cell migration rate=[experimental group cell number meansigma methods/blank group cell number meansigma methods] × 100%.
The effect that result display sulfamethoxazole compound has the suppression of dose dependent to move to colon cancer cell HT29, compares migrating cell relative percentages and significantly reduces (P < 0.05) with matched group.In table 2.
Table 2 Transwell method detects sulfamethoxazole to the impact (n=3x ± s) of HT29 cell migration
Embodiment 3: injury repairing experiment detects sulfamethoxazole to tumor cell migration capacity.
Take the logarithm the HT29 cell of trophophase, in 24 orifice plates, every hole adds cell 1 × 10
5, grow up to after monolayer until cell, draw " one " font cut, PBS rinses, and matched group and experimental group add sulfamethoxazole variable concentrations respectively and take pictures, and after cultivating 22h, take pictures again in same position, measures migration distance.Experiment repetition 3 times, cell migration rate=[(experimental group 0h meansigma methods-experimental group 22h meansigma methods)/(blank group 0h meansigma methods-blank group 22h meansigma methods)] × 100%.
Sulfamethoxazole (10-100 μ g/ml) under variable concentrations detects the suppression migration after LTEP, A549 cytosis 22h, the effect that result display sulfamethoxazole has the suppression of dose dependent to move to colon cancer cell HT29.Compare cell mobility significantly to reduce (P < 0.05) with matched group.In table 3.
The experiment of table 3 injury repairing detects sulfamethoxazole to colon cancer cell HT29 mobility impact (n=3x ± s)
Conclusion: sulfamethoxazole significantly can suppress the migration of colon cancer cell, can be used for inhibitor against colon carcinoma cells diversion medicaments of curing the disease.
As mentioned above, although represented with reference to specific preferred embodiment and described the present invention, it shall not be construed as the restriction to the present invention self.Under the spirit and scope of the present invention prerequisite not departing from claims definition, various change can be made in the form and details to it.
Claims (3)
1. the application of sulfamethoxazole in the medicine preparing inhibition tumor cell metastasis and extension.
2. application according to claim 1, wherein tumor cell is colon cancer cell.
3. application according to claim 1, wherein the concentration of sulfamethoxazole is 50 ~ 60uM.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2019500382A (en) * | 2015-12-31 | 2019-01-10 | キョンブク ナショナル ユニバーシティ インダストリー−アカデミック コーオペレーション ファウンデーション | Pharmaceutical composition for treating cancer and inhibiting metastasis comprising a sulfonamide compound as an active ingredient |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1729012A (en) * | 2002-10-24 | 2006-02-01 | 伊利诺伊大学评议会 | Compositions and methods for preventing and treating solid tumors |
| US20100285012A1 (en) * | 2008-01-05 | 2010-11-11 | University Of Florida Research Foundation Inc. | Methods and compositions for the treatment of cancers and pathogenic infections |
| CN102460165A (en) * | 2009-05-19 | 2012-05-16 | 维维雅生物技术公司 | Methods for providing personalized medicine tests ex vivo for hematological neoplasms |
| CN103945847A (en) * | 2011-12-02 | 2014-07-23 | 信一佰奥建有限公司 | Complex antibacterial composition for animals |
-
2015
- 2015-02-09 CN CN201510068148.3A patent/CN104606207A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1729012A (en) * | 2002-10-24 | 2006-02-01 | 伊利诺伊大学评议会 | Compositions and methods for preventing and treating solid tumors |
| US20100285012A1 (en) * | 2008-01-05 | 2010-11-11 | University Of Florida Research Foundation Inc. | Methods and compositions for the treatment of cancers and pathogenic infections |
| CN102460165A (en) * | 2009-05-19 | 2012-05-16 | 维维雅生物技术公司 | Methods for providing personalized medicine tests ex vivo for hematological neoplasms |
| CN103945847A (en) * | 2011-12-02 | 2014-07-23 | 信一佰奥建有限公司 | Complex antibacterial composition for animals |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2019500382A (en) * | 2015-12-31 | 2019-01-10 | キョンブク ナショナル ユニバーシティ インダストリー−アカデミック コーオペレーション ファウンデーション | Pharmaceutical composition for treating cancer and inhibiting metastasis comprising a sulfonamide compound as an active ingredient |
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Application publication date: 20150513 |