CN106692152A - Application of GSK503 in preparing drug for treating RB (Retinoblastoma) - Google Patents
Application of GSK503 in preparing drug for treating RB (Retinoblastoma) Download PDFInfo
- Publication number
- CN106692152A CN106692152A CN201710031936.4A CN201710031936A CN106692152A CN 106692152 A CN106692152 A CN 106692152A CN 201710031936 A CN201710031936 A CN 201710031936A CN 106692152 A CN106692152 A CN 106692152A
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- China
- Prior art keywords
- gsk503
- retinoblastoma
- treating
- drug
- treatment
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- XOZKQIPQTXLJIH-UHFFFAOYSA-N CCC(CC)C(NC)=C Chemical compound CCC(CC)C(NC)=C XOZKQIPQTXLJIH-UHFFFAOYSA-N 0.000 description 1
- 0 C[C@@](CC(C)=C(C)CC*)N Chemical compound C[C@@](CC(C)=C(C)CC*)N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene
Abstract
The invention belongs to the field of treating RB (Retinoblastoma), and particularly relates to application of GSK503 in preparing a drug for treating the RB. In the drug, the final concentration of the GSK503 is greater than or equal to 1 mu M. The GSK503 is a novel EZH2 (Enhancer of Zeste Homolog 2) small-molecule inhibitor, and an applicant experimentally finds that the GSK503 is capable of remarkably inhibiting the growth of the RB, and the methylation degree of H3K27me3 in cells is reduced through inhibiting the EZH2, so that cancer suppressor genes can be in re-expression, and the effect of inhibiting the tumor growth is reached. Effective treatment concentration can be reached when the final concentration of the GSK503 reaches 1 mu M, the use effect of the drug is increased while the concentration is increased, and the growth of the RB can be remarkably inhibited.
Description
Technical field
The invention belongs to the therapy field of retinoblastoma, more particularly to a kind of GSK503 is preparing treatment retina
Application in blastoma medicine.
Background technology
Retinoblastoma (Retinoblastoma, RB) is the most common intraocular malignant of children, often in development
During develop, be a kind of embryonic malignant tumor for originating from retina stratum nucleare.The tumour has Family inherited inclination,
It is property most serious, a kind of malignant tumour of harmfulness maximum in infant's illness in eye, its Vision prognosis is poor, and is susceptible to cranium
Interior and DISTANT METASTASES IN, often jeopardizes patient vitals.
At present, the treatment for the tumour is mainly the non-etiological treatments such as operation and chemicotherapy, lacks for its cause of disease
Treatment means.Zeste genetic enhancers homologue 2 (EZH2) is one of members for combing protein complexes (PcG) families more, is many
Comb albumen suppresses the catalysis activity subunit of complex 2 (PRC2).EZH2 is a kind of intracellular ZNFN3A1, it
Methylating for trimethyl (H3K27me3) on the 27th amino acid of intracellular histone H 3 can be promoted, and be proved to be a kind of former
Oncogene, the growth and transfer to kinds of tumors is related.EZH2 can be suppressed by improving H3K27me3 methylation levels in cell
Expression of tumor suppressor gene, so as to cause tumour to occur.GSK503 is a kind of new EZH2 micromolecular inhibitors, and it is proved in skin
The growth and transfer of tumour can be significantly inhibited in skin malignant mela noma, and Normal melanocytes are not made significant difference, thus
It is considered as having preferable prospect to oncotherapy.But so far, still no GSK503 is applied in retinoblastoma
Report, its curative effect and mechanism of action to retinoblastoma is unclear.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of GSK503 and are preparing treatment retinoblastoma medicine
In application, the invention improves treatment it is intended that the clinical treatment of retinoblastoma provides new target spot and medicine
Validity, reduces eyeball excise rate, improves patient visual's prognosis.
Applications of the GSK503 of the invention in treatment retinoblastoma medicine is prepared, chemical structural formula such as Formulas I institute
Show,
Preferably, GSK503 final concentrations are more than or equal to 1 μM in described medicine.
Preferably, final concentration of 1-10 μM of GSK503 in described medicine.
Preferably, final concentration of 1,5 or 10 μM of GSK503 in described medicine.
GSK503 is a kind of new EZH2 micromolecular inhibitors, and molecular formula is C31H38N6O2, molecular weight 526.67g/
Mol, molecular structural formula (shown in formula I).Applicant's experiment finds that GSK503 can significantly inhibit the life of retinoblastoma
It is long, by suppressing EZH2, make intracellular H3K27me3 methylations reduction, so that tumor suppressor gene is expressed again, reach suppression
The effect of tumour growth processed.
Beneficial effect：
GSK503 final concentrations reach 1 μM and can reach effective treatment concentration, and drug effectiveness with concentration rising
And increase, retinoblastoma growth can be significantly inhibited.
Brief description of the drawings
Fig. 1 is the experimental result picture that RB tissues are carried out immunofluorescence with emmetropia tissue.
Fig. 2 is chromatin imrnunoprecipitation experimental result picture, and wherein A is RB44 compared with control cells and the RB44 with GS503 treatment
The chromatin imrnunoprecipitation PCR result figures of versus, B is the quantitative analysis results figure of A figures.
Fig. 3 is proliferation experiment result figure of the RB44 cells after different final concentration GS503 treatment.
Specific embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than limitation the scope of the present invention.In addition, it is to be understood that after the content for having read instruction of the present invention, people in the art
Member can make various changes or modifications to the present invention, and these equivalent form of values equally fall within the application appended claims and limited
Scope.
Embodiment 1
Immunofluorescence experiment
Experiment material：Retinoblastoma by tissue microarray (contains 28 Tissues of Retinoblastoma, 12 emmetropia groups
Knit), purchased from Xi'an Ai Li bio tech ltd；H3K27me3 antibody and SA, purchased from Abcam (Ai Bokang (on
Sea) trade Co., Ltd).
Organization chip is placed in wet box, paving plus PBS are on organization chip.The H3K27me3 antibody of dilution in 4 DEG C,
13500g is centrifuged 2min.The PBS on slide is sucked in one end of organization chip, and antibody is added from the other end, covered
Humidification box, incubation at room temperature 1h.With Xian's PBS slide 3 times (5min/ times), new PBS is added from section one end, by another
End sucks old buffer solution.The SA of dilution is in 4 DEG C, 13500g centrifugations 2min.SA is added on organization chip,
Incubation at room temperature 1h in humidification box is put, slide 3 times (5min/ times) is washed with PBS.Lid cover glass, basis of microscopic observation result is (see figure
1).As can be drawn from Figure 1, H3K27me3 immunofluorescence dyeings are carried out by the organization chip to retinoblastoma, is demonstrate,proved
H3K27me3 methylations are compared normal structure and are significantly raised in real Tissues of Retinoblastoma.
Embodiment 2
Chromatin immunoprecipitation assay (ChIP) is tested
Experiment material：Human retinoblastoma cell line RB44 cells, 37 DEG C, 5%CO2Cellar culture, culture medium is
The RPMI-1640 (Gibco) of 10% hyclone (FBS)；GSK503 is purchased from MedChemExpress (MCE China)；ChIP is tried
Agent box is purchased from millipore (U.S. Mi Libo)；PCR kit is purchased from Takara (precious bioengineering (Dalian) Co., Ltd)
RB44 compared with control cells and cell (10 μM of GSK503,37 DEG C, 5%CO of GS503 treatment2Cellar culture 7 days) 800g
Centrifugation 5min, abandons supernatant, and 10ml is resuspended for the training liquid of 10%FBS 1640, adds the μ l of 37% formalin 270, is incubated at room temperature 10min
After add 600 μ l 2M glycine terminating reactions, in 4 DEG C, 2500g centrifugation 15min are cracked after abandoning supernatant with cell pyrolysis liquid.It is super
Sound 5min smashes DNA, and in 4 DEG C, 13500g is centrifuged 15min, and take supernatant carries out ChIP realities according to ChIP kits operating procedure
Test, albumen and its DNA fragmentation of combination are left behind with H3K27me3 and EZH2 antibody, after removing removing protein, add two volumes
Minus 20 degree of the sodium acetate of absolute ethyl alcohol and 10% cumulative volume is precipitated DNA2 hours, in 4 DEG C, 13500g centrifugation 20min, 75% second
Product is obtained after alcohol washing, the combination situation (see Fig. 2) of H3K27me3, EZH2 and target dna is detected by PCR.Can from Fig. 2
Combined with the promoter region of tumor suppressor gene RB1 genes with learning that GSK503 can suppress EZH2, made its promoter region H3K27me3 first
The reduction of base degree.
Embodiment 3
Cell proliferation experiment (CCK8)
Experiment material：CCK8 is purchased from Japanese colleague's chemistry
RB44 cells train liquid, 37 DEG C, 5%CO with the 10%FBS 1640 containing final concentration of 10 μM of GSK5032Cellar culture 7
My god, the cell after treatment and control group RB4 cells are inoculated in 96 orifice plates, per 5000, hole cell, 100ul training liquid, 37 DEG C,
5%CO2Culture, respectively at 0h, 24h, 48h, 72h adds 10ul CCK8, continues 37 DEG C of incubation 4h, upper machine survey absorbance (see
Fig. 3).As can be known from Fig. 3, GSK503 can significantly inhibit the propagation of retinoblastoma, and inhibition with concentration rising
Enhancing.
Claims (4)
- Applications of the 1.GSK503 in treatment retinoblastoma medicine is prepared, such as chemical structural formula of the GSK503, Formulas I It is shown,
- 2. applications of the GSK503 according to claim 1 in treatment retinoblastoma medicine is prepared, its feature exists In GSK503 final concentrations are more than or equal to 1 μM in described medicine.
- 3. applications of the GSK503 according to claim 2 in treatment retinoblastoma medicine is prepared, its feature exists In final concentration of 1-10 μM of GSK503 in described medicine.
- 4. applications of the GSK503 according to claim 2 in treatment retinoblastoma medicine is prepared, its feature exists In final concentration of 1,5 or 10 μM of GSK503 in described medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710031936.4A CN106692152A (en) | 2017-01-17 | 2017-01-17 | Application of GSK503 in preparing drug for treating RB (Retinoblastoma) |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710031936.4A CN106692152A (en) | 2017-01-17 | 2017-01-17 | Application of GSK503 in preparing drug for treating RB (Retinoblastoma) |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106692152A true CN106692152A (en) | 2017-05-24 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710031936.4A Pending CN106692152A (en) | 2017-01-17 | 2017-01-17 | Application of GSK503 in preparing drug for treating RB (Retinoblastoma) |
Country Status (1)
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| CN (1) | CN106692152A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020011607A1 (en) * | 2018-07-09 | 2020-01-16 | Fondation Asile Des Aveugles | Inhibition of prc2 subunits to treat eye disorders |
| CN110755437A (en) * | 2019-09-16 | 2020-02-07 | 温州医科大学 | Application of SYK inhibitor R406 and BAY-61-3606 in preparation of medicine for treating retinoblastoma |
| CN117296799A (en) * | 2023-11-28 | 2023-12-29 | 四川省医学科学院·四川省人民医院 | Construction method and application of retinal pigment degeneration disease model |
-
2017
- 2017-01-17 CN CN201710031936.4A patent/CN106692152A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| CHANGPING WU ET AL.: "Inhibition of EZH2 by chemo- and radiotherapy agents and small molecule inhibitors induces cell death in castration-resistant prostate cancer.", 《ONCOTARGET》 * |
| MEHNAZ KHAN ET AL.: "Characterization and pharmacologic targeting of EZH2,a fetal retinal protein and epigenetic regulator, inhuman retinoblastoma.", 《LABORATORY INVESTIGATION》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020011607A1 (en) * | 2018-07-09 | 2020-01-16 | Fondation Asile Des Aveugles | Inhibition of prc2 subunits to treat eye disorders |
| CN112399857A (en) * | 2018-07-09 | 2021-02-23 | 盲人庇护基金会 | Inhibition of PRC2 subunit for the treatment of ocular disorders |
| US20210213013A1 (en) * | 2018-07-09 | 2021-07-15 | Fondation Asile Des Aveugles | Inhibition of prc2 subunits to treat eye disorders |
| CN110755437A (en) * | 2019-09-16 | 2020-02-07 | 温州医科大学 | Application of SYK inhibitor R406 and BAY-61-3606 in preparation of medicine for treating retinoblastoma |
| CN117296799A (en) * | 2023-11-28 | 2023-12-29 | 四川省医学科学院·四川省人民医院 | Construction method and application of retinal pigment degeneration disease model |
| CN117296799B (en) * | 2023-11-28 | 2024-02-02 | 四川省医学科学院·四川省人民医院 | Construction method and application of retinal pigment degeneration disease model |
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| PB01 | Publication | ||
| PB01 | Publication | ||
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| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170524 |