CN108125974A - Application of the tilianin and combinations thereof in anti-angiogenic medicaments are prepared - Google Patents
Application of the tilianin and combinations thereof in anti-angiogenic medicaments are prepared Download PDFInfo
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Abstract
The invention discloses a kind of application of the tilianin and combinations thereof in anti-angiogenic medicaments are prepared, the anti-angiogenic medicaments are tilianin and combinations thereof.Tilianin of the present invention and combinations thereof is in the application in preparing anti-angiogenic medicaments, there is tilianin anti-angiogenesis activity, tilianin and combinations thereof can be used as angiogenesis inhibitors treatment and the relevant disease of angiogenesis such as tumour, arthritis, atherosclerosis etc..
Description
Technical field
The present invention relates to pharmaceutical technology field, specifically, be related to a kind of tilianin and combinations thereof prepare it is anti-angiogenic
Generate the application in drug.
Background technology
Blood vessel spreads all over human body, is first organ in embryo, they carry oxygen to providing energy for organ growth everywhere
Amount.Human body major part blood vessel keeps quiescent condition in most of time after adult, and angiogenesis is only in placenta, female pathology week
Interim generation.Many stimulating factors and inhibiting factor such as oxygen, integrin, Endostatin etc. in human body environment are kept well
This physiological equilibrium.When human body encounters wound, during anoxic, this balance is broken, and endothelial cell rapidly makees these stimulations
Go out reaction.Some reactions are beneficial, such as wound reparation and healing, but excessive vascular dysregulation will lead to disease, such as
Cancer, autoimmune disease, vascular malformation, DiGeorge syndromes, obesity, psoriasis, diabetic retinopathy, heavy breathing
Asthma, synovitis etc.；And Alzheimer disease, atherosclerosis, skin purpura, nephrosis and osteoporosis feature or the cause of disease make
Angiogenesis is insufficient or blood vessel regression.
Malignant tumour be now human death the main reason for one of.The origin of malignant tumour is not angiogenesis, still
Tumour is reaching 1mm3When, if no blood vessel provides enough nutrition tumours and will be unable to continued growth and transfer.In addition, swollen
In the presence of knurl, the secretion of various growth factors can sharply increase, and promote angiogenesis, such as VEGF, EGF, FGF, PDGF.
Wherein most representative is VEGF, it is considered as the first triggering factors of angiogenesis.Many activity dues are in VEGF energy
Enough activating phosphatase inositol 3-kinase (PI3 kinases)-Akt and MAPK signal paths, this two accesses and cell Proliferation are migrated, are invaded
It attacks etc. closely related.Scientists are dedicated to studying any anti-angiogenesis that can inhibit above-mentioned one or more approach for many years
Drug is to inhibit the growth of tumour and transfer.Unfortunately, many targeted drugs can be very good that tumor cell in vitro is inhibited to increase
It grows and is shifted with the tumour growth of tumor animal, but therapeutic effect clinically is unsatisfactory.Foremost is Avastin
(Roche), the combination of VEGF and VEGFRs is targeted, is only combined on the basis of classic chemotherapy medicine, colorectal cancer and non-small cell
Patients with lung cancer can clinical Benefit, while concurrent many adverse reactions again.
In conclusion single anti-angiogenesis strategy may cause drug resistance, drug effect insufficient, it is badly in need of finding multipath suppression
Angiogenesis processed generates smaller drug resistance, lower-cost drug.
Compared with chemical synthetic drug, the compound detached from natural plants is also the good source of new drug.Dracocephalum moldavica is
Uygur nationality's traditional herbal medicines containing tens of kinds of flavone compounds, are mainly used for treating breathing problem, neurasthenia and painstaking effort
Pipe disease.Flavonoids drug generally has the effects that anti-inflammatory, antiviral, antitumor, anti-oxidant, protection liver.Tilianin
(Tilianin, TIL) be Dracocephalum moldavica active constituent flavones in main component, molecular formula C22H22O10, molecular weight 446,
The chemical structural formula of tilianin is as follows：
Have the study found that activity of the tilianin with anti-diabetic, reducing blood lipid；TGF-β/Smad signals can be lowered to lead to
Road；Lower EGFR-MEK-Erk and Akt signal paths；Inhibit iNOS expression (Nam KH, Choi JH, Seo YJ, et
al.Inhibitory effects of tilianin on theexpression of inducible nitric oxide
synthase in low density lipoprotein receptordeficiency mice.Exp Mol
Med.2006Aug 31；38(4):445-52).But tilianin is in preparing inhibition angiogenesis, inhibiting tumour growth drug
Application have not been reported.
Invention content
The object of the present invention is to provide a kind of application of the tilianin and combinations thereof in anti-angiogenic medicaments are prepared.
To achieve these goals, the technical solution adopted by the present invention is as follows：
The present invention provides a kind of application of the tilianin and combinations thereof in anti-angiogenic medicaments are prepared.
The anti-angiogenic medicaments are tilianin and combinations thereof.
The angiogenesis is neonate tumour blood vessel, psoriatic lesions tissue blood vessel is newborn, Paget's disease blood vessels are new
Blood at life, the angiogenesis of non-malignant vascular proliferative disease, the angiogenesis of arthritis pathological tissues, atherosclerotic lesion
The angiogenesis of pipe new life or neovascular eye diseases.
The neonate tumour blood vessel is angiogenesis caused by the angiogenesis or tumour of tumor tissues.
The neovascular eye diseases are primary or secondary neovascular eye diseases.
The tumour is primary or secondary entity tumor.
The dosage form of the drug is injection, capsule, tablet, powder, granule, pill, microcapsules and microsphere preparation, bolt
Agent, ointment, spray or targeting preparation.
The administering mode of the drug is oral, injection.
The anti-angiogenic medicaments refer to tilianin single component as drug or creeping thistle for tilianin and combinations thereof
Glycosides forms composition with other pharmaceutically acceptable ingredients.
Other described pharmaceutically acceptable ingredients are that do not have the drug of antagonism with tilianin or pharmaceutically allow
One or more auxiliary materials.
The tilianin is formed with other pharmaceutically acceptable ingredients in composition, and the weight of active component tilianin contains
It measures as 0.1-99%.
The chemical structural formula of the tilianin is：
Due to the adoption of the above technical scheme, the present invention has the following advantages and beneficial effect：
In the application in preparing anti-angiogenic medicaments, tilianin has to be inhibited tilianin of the present invention and combinations thereof
The effect of tumour growth, Antineoplastic angiogenesis, tilianin and combinations thereof can be used to treat or prevent and tumour or blood vessel life
Into relevant disease.Meanwhile tilianin is flavonoids drug, the low tolerance of toxicity is good, the big portion with generating larger side effect
Antitumor drug is divided to compare, there is apparent advantage.
Tilianin of the present invention and combinations thereof finds tilianin for the first time in the application in preparing anti-angiogenic medicaments
The proliferation of huve cell (HUVECs), migration, tubule under VEGF-A inductions can be significantly inhibited to be formed, find field for the first time
Ji glycosides can induce huve cell apoptosis, finds that tilianin can significantly inhibit angiogenesis in Mice Body for the first time, sends out for the first time
Existing tilianin can inhibit the growth of nude mice by subcutaneous inoculated tumour by blood vessel formation against function, find that tilianin can block blood for the first time
Endothelial tube growth factor acceptor 2 (VEGFR2) signal path and its downstream protein kinase B (Akt) signal path, extracellular adjusting
The phosphorylation of protein kinase (Erk1/2) signal path, so as to play the effect of its anti-angiogenesis；It can be seen that tilianin has
There is anti-angiogenesis activity, tilianin and combinations thereof that can be used as angiogenesis inhibitors treatment and the relevant disease of angiogenesis
Such as tumour, arthritis, atherosclerosis.
Description of the drawings
Fig. 1 is that tilianin inhibits the HUVECs proliferation (A) of VEGF-A inductions, the result schematic diagram of HUVECs migrations (B).
Fig. 2 is the result schematic diagram that tilianin inhibits the HUVECs tubules of VEGF-A inductions to be formed；Wherein A is fluorescent staining
Figure；B is the quantitative analysis figure by ImageJ softwares.
Fig. 3 is the result schematic diagram that tilianin inhibits angiogenesis in the Mice Body of VEGF-A inductions；Wherein A is model group
(plug1), VEGF-A stimulations group (plug2) and tilianin treatment group (plug3) matrigel bolt representativeness picture；B is model group
VEGF-A stimulations group and the representative picture of tilianin treatment group matrigel bolt HE dyeing；C is model group, VEGF-A stimulations group and field
Quantitative hemoglobin in Ji Gan treatment groups matrigel；D exempts from for model group, VEGF-A stimulations group and tilianin treatment group matrigel bolt
Epidemic disease group CD31 dyes picture；E is model group, VEGF-A stimulations group and tilianin treatment group matrigel immunohistochemistry CD31 dye
Microvessel density statistical analysis afterwards.
Fig. 4 is the result schematic diagram that tilianin inhibits A549 tumor-bearing mice tumour growths by anti-angiogenesis；Wherein A is
Model group and tilianin treatment group tumor-bearing mice weight statistics；B is model group and tilianin treatment group tumors volume statistical analysis
And tumor tissues picture；C is model group and tilianin treatment group tumors histogenic immunity group CD31 dyeing and statistical analysis；D is
Model group and tilianin treatment group tumors tissue T unel dyeing (tumour cell of apoptosis) and statistical analysis.
Fig. 5 is showing for the tilianin VEGFR2 that is induced through VEGF-A of inhibition and downstream signaling pathway Akt, Erk1/2 phosphorylation
It is intended to.
Specific embodiment
In order to illustrate more clearly of the present invention, with reference to preferred embodiment, the present invention is described further.Ability
Field technique personnel should be appreciated that following specifically described content is illustrative and be not restrictive, this should not be limited with this
The protection domain of invention.
Tilianin used flies bio tech ltd, lot number E218392, specification purchased from Shanghai one in the following example
1g。
Test method without specific conditions in the following example, usually according to normal condition, reagent used from
Market is commercially available.
Embodiment 1
Tilianin inhibits HUVEC proliferation experiments
Purpose and principle：Using the proliferation of CCK-8 methods detection human umbilical vein endothelial cell (HUVEC).CCK-8 methods are
It is molten for measuring number of viable cells or a kind of highly sensitive, "dead" colorimetric detection method of cell Proliferation in toxicity test
WST-8 can be dehydrogenated enzyme reduction and generate orange-yellow formazan in the presence of electron carrier 1-Methoxy PMS in liquid
Product, the depth (orange) of solution colour is directly proportional to the proliferation of cell to be measured, the absorbance OD measured at 450nm wavelength
Value can reflect the quantity of living cells indirectly to a certain extent.
Method：Using CCK-8 kits, when HUVECs grows to 80%, with trypsin digestion, then by HUVECs
(5000 cells/well/100uL) is inoculated into 96 orifice plates, at 37 DEG C, 5%CO2Lower incubation.After 24 hours, with containing different dense
The fresh complete medium of the tilianin (0,10,20,40,80,160 μM) of degree replaces former culture medium, and continues culture 24 hours
Or 48 hours.In the experiment of VEGF-A stimulations, by cell inoculation in 96 orifice plates, after 24 hours, with or without VEGF-A
The tilianin of (15ng/ml) and various concentration (0,10,20,40,80,160 μM) continues to be incubated 24 or 48 hours, then uses PBS
Each hole is washed, adds in the 100 μ L fresh cultures containing 10uL CCK-8 reagents, after being incubated 1.5 hours, is measured using microplate reader
The absorbance of each sample at 490nm.
As a result with conclusion：The results are shown in Figure 1, and Fig. 1 is that tilianin inhibits the HUVECs proliferation of VEGF-A inductions, HUVECs
The result schematic diagram of migration；In Figure 1A, tilianin can significantly inhibit the HUVECs proliferation of VEGF-A inductions, dosage and time be according to
Lai Xing.(a concentration of 0) of tilianin, VEGF-A can significantly induce HUVECs to be proliferated (P compared with blank control group<0.01)；With
VEGF-A inductions group is compared to (tilianin of 10-160 μM of a concentration of 0) of tilianin is 4.26%- to the inhibiting rate of HUVECs for 24 hours
77.33%, it is 5.95%-81.91% (* in 48h:P<0.05；**:P<0.01).
Embodiment 2
Tilianin inhibits HUVEC migration experiments
Purpose and principle：Transwell experimental techniques, the major experimental tool of this experiment is Transwell cells.Experiment
In, cell is placed on cell, containing under the action of growth factor under cell, the polycarbonate membrane of cell permeable cell bottom into
The experimental study of row cell migration.
Method：With the HUVECs (2.1 × 10 of (0,10,20,40 μM) 6 orifice plates of processing of tilianin5/ hole) 24 hours, then
Every group with 200ul serum free mediums by processed cell (2 × 104) cell of Transwell is seeded in, lower room is
600ul is with or without the culture medium of the 1%FBS of VEGF-A (15ng/ml).It is thin with cotton swab upper chamber after migration 16 hours
Born of the same parents fix cell 20 minutes below cell film with 100% methanol at room temperature, are dyed 2 minutes with DAPI, be inverted with Olympus
Every group of random shooting 5 of microscope opens the visual field (200 times of amplifications), and cell migration ability is assessed by counting staining cell.
As a result with conclusion：The results are shown in Figure 1, and Fig. 1 is that tilianin inhibits the HUVECs proliferation of VEGF-A inductions, HUVECs
The result schematic diagram of migration；In Figure 1B, (a concentration of 0) of tilianin, VEGF-A can improve HUVECs's compared with blank control group
Transfer ability (90%, p<0.01)；But (a concentration of 0) of tilianin, tilianin can significantly inhibit compared with VEGF-A induction groups
The HUVECs migrations of VEGF-A inductions, 40 μM of tilianin can inhibit the HUVECs migrations (* that 87.15%VEGF-A is induced:P<
0.05；**:P<0.01).
Embodiment 3
Tilianin inhibits HUVECs to be tested into pipe
Purpose and principle：Artificial basement membrane Matrigel glue is the basement membrane component extracted from mouse EHS sarcomas, at 37 DEG C
Gelatinous Artificial Basement Membrane can be spontaneously formed, the biological action with natural basement membrane.HUVECs can glue on Matrigel glue
It is attached into pipe, so as to be used to study the influence of drug Human Umbilical Vein Endothelial Cells extracorporeal blood vessel nucleus formation.
Method：It will be positioned at low 4 DEG C of growth factor matrigel and stay overnight on ice, suction pipette head used and 24 orifice plates is pre-
It is cold, then 250 μ L matrigels are added in each hole of 24 porocyte culture plates on ice, are gently shaken to prevent bubble,
3 hours are subsequently placed in 37 DEG C of cell incubator to form gel.In first method, by the HUVECs of overnight starvation
(105A/hole) inoculation contain or not contain at the same time VEGF-A (15ng/ml) and various concentration tilianin (0,10,20,
40 μM) processing matrigel covering hole in.After 6-8 hours, culture medium is replaced with containing 30 μ L calceins solution (10 μ
M 300 μ L culture mediums), and continue to be incubated 20 minutes.Every group of inverted microscope (Olympus) is used under 40 times of amplification factors
5 visuals field of random shooting, and with ImageJ softwares quantitative analysis is carried out by counting into pipe branch point, into pipe number, into pipe area.
As a result with conclusion：The results are shown in Figure 2, and Fig. 2 is the HUVECs tubules formation that tilianin inhibits VEGF-A inductions
Result schematic diagram；In Fig. 2A, (a concentration of 0) of tilianin, HUVECs can be spontaneously formed on matrigel compared with blank control group
Tubular structure, and VEGF-A induction groups form tubule (P more, that structure is thicker<0.01), and tilianin is with the side of concentration dependant
The pipe that formula can block VEGF-A inductions is formed.10 μM of tilianin can inhibit tubule formed (>40% inhibits), 40 μM of tilianin
Pipe can almost be completely inhibited and form (*:P<0.05；**:P<0.01)；Fig. 2 B are that each group in Fig. 2A is regarded by ImageJ softwares
Open country carries out the result schematic diagram of quantitative analysis into pipe into pipe branch point, into pipe number, into pipe area.
Embodiment 4
Tilianin inhibits body vessel generation experiment
Purpose and principle：Artificial basement membrane Matrigel glue is the basement membrane component extracted from mouse EHS sarcomas, at 37 DEG C
Gelatinous Artificial Basement Membrane can be spontaneously formed, the biological action with natural basement membrane.Endothelial cell in Mice Body is transportable
To internal Matrigel glue has been implanted into, blood vessel is formed in Matrigel glue, it is thin to endothelium so as to be used to study drug
The influence of angiogenesis function in cell space.
Method：Matrigel and syringe are placed on ice bag before injection, it will be with 20U heparin or 200ng VEGF-A or field
The matrigel (0.5ml) of Ji glycosides (120mg/kg) mixing is subcutaneously implanted in the right armpit of 5-6 week old male nude mouse (6 mouse/groups)
Under, negative control group only contains heparin.12 days after implantation, mouse is put to death with chloraldurate, takes out the matrigel of implantation.Matrigel
Measure the content of hemoglobin in every group of matrigel with microplate reader at 530nm using hemoglobin detection kit.Part base
CD31 dyeing calculates microvessel density after matter glue bolt paraformaldehyde fixes paraffin embedding.
As a result with conclusion：The results are shown in Figure 3, and Fig. 3 is that tilianin inhibits angiogenesis in the Mice Body of VEGF-A inductions
Result schematic diagram；It is model group (plug1), VEGF-A stimulations group (plug2) and tilianin treatment group (plug3) base in Fig. 3 A
Matter glue bolt representative schematic diagram；Model group, VEGF-A stimulations group and tilianin treatment group matrigel bolt HE dyes are followed successively by Fig. 3 B
Color representative schematic diagram；It is model group in Fig. 3 C, quantitative hemoglobin in VEGF-A stimulations group and tilianin treatment group matrigel
Schematic diagram；It is model group, VEGF-A stimulations group and tilianin treatment group matrigel bolt immunohistochemistry CD31 dyeing in Fig. 3 D
Schematic diagram；It is micro- blood after model group, VEGF-A stimulations group and tilianin treatment group matrigel immunohistochemistry CD31 are dyed in Fig. 3 E
The schematic diagram of pipe Statistics of Density analysis；It is consistent with extracorporeal blood vessel generation experimental result, with negative control group (containing only 20U heparin, bolt
1) plug is compared, 200ng VEGF-A (plug2) can dramatically increase angiogenesis in internal matrigel, and tilianin group (contains 200ng
VEGF-A, 20U heparin, 120mg/kg tilianins, plug3) significantly reverse VEGF-A induction angiogenesis.VEGF-A induction groups
Hemoglobin (Hb) is higher than control group 87%, and tilianin group Hb contents reduce 81% (* compared with VEGF-A induction groups:P<
0.05；**:P<0.01).
Embodiment 5
Tilianin inhibits A549 tumor-bearing mice tumour growths by anti-angiogenesis
Purpose and principle：By observing influence of the tilianin to the growing state of tumor-bearing mice tumour, passing through immunohistochemistry
Microvessel density in CD31 staining analysis tumor tissues passes through tumour cell in Tunel experimental analysis tumor-bearing mice tumor tissues
Apoptosis situation, overall merit tilianin is by inhibiting the antitumor action of angiogenesis.
Method：1~2 week predetermined nude mice, makes nude mice fully adapt to experimental situation before experiment.Cultured and amplified in vitro A549 is thin
Born of the same parents, the A549 cells total amount according to needed for determining the number of experiment mice.It is good and in exponential phase to choose growth conditions
A549 cells, are digested with trypsase, and machinery piping and druming is washed 2 times into single cell suspension, and with PBS, and action is soft, keeps away
Exempt from clasmatosis, centrifugation cell.It is resuspended with the DMEM culture solutions of serum-free, 20 μ L cell suspensions is taken to be counted, it will be thin
Born of the same parents' concentration is adjusted to 5 × 107A/mL.1.5mL EP pipes are sub-packed under aseptic condition, often pipe dispenses 100 μ L, i.e., often pipe comprising 5 ×
106A cell.With 1mL asepsis injectors by A549 cell inoculations, every mouse inoculation 5 × 10 subcutaneous in the right oxter of nude mice6It is a thin
Born of the same parents (100 μ L of volume), then conventinal breeding nude mice.The growth conditions of observation mouse and the upgrowth situation of tumour daily, 4 days
Afterwards, for mouse subcutaneously into knurl, knurl product is about 80mm3, 30 experiment nude mices are randomly divided into 3 groups：Control group, tilianin low dosage
Group, tilianin high dose group, every group 10.Pharmaceutical intervention is given to three groups of mouse, wherein control group tumor-bearing mice is given daily
Physiological saline containing DMSO, low dose group give the tilianin of 30mg/kg daily, and high dose group gives the field of 80mg/kg daily
Ji glycosides, is administered by the way of intraperitoneal injection.Every other day with the major diameter (a) and minor axis of vernier caliper measurement subcutaneous transplantation knurl twice
(b), the volume of tumour is calculated：V(mm3)=0.5a × b2, draw growth of transplanted human curve.Successive administration 15 days, in last time
Mouse is put to death after administration 6h, bat takes animal and tumour photo, and tumor tissues are carried out paraffin embedding histochemical staining, Tunel, analysis
Apoptosis cell in microvessel density, tumor tissues in tumor tissue growth's trend, tumor tissues.
As a result with conclusion：The results are shown in Figure 4, and Fig. 4 is that tilianin inhibits A549 tumor-bearing mices to swell by anti-angiogenesis
The result schematic diagram of knurl growth；It is tumor-bearing mice weight statistical result in model group and tilianin treatment group in Fig. 4 A, in Fig. 4 B
It is model group and field for model group and tilianin treatment group tumors volume statistical analysis and tumor tissues picture schematic diagram, in Fig. 4 C
Ji glycosides treatment group tumors histogenic immunity group CD31 is dyed and statistical analysis schematic diagram, is treated in Fig. 4 D for model group and tilianin
Group tumor tissues Tunel dyeing (tumour cell of apoptosis) and statistical analysis schematic diagram；The weight of the mouse of three groups is not poor
It is different, show apparent side effect of the tilianin without result in weight loss, and mouse can be resistant to the field of low dosage and high dose
The processing of Ji glycosides.As shown in Figure 4 B, gross tumor volume continues at any time, and at the 19th day, control group gross tumor volume reached average 535mm3,
Low dose group reaches average 515mm3, and high dose group only averagely 48mm3, prompt high dose tilianin that can significantly inhibit swollen
Knurl grows (P<0.001).Tumour CD31 dyeing quantitative analyses show that tilianin high dose group is to Tumor Angiongesis inhibiting rate height
Up to 71% (P<0.01) no significant difference (Fig. 4 C), and between control group and low dose group；TUNEL detection displays, with
Control group is compared, and high dose tilianin can significantly induce the apoptosis (Fig. 4 D) of tumor tissues.
Embodiment 6
Tilianin inhibits VEGFR2 signal paths in HUVECs
Purpose and principle：Angiogenesis is carried out by multiple steps, including endothelial cell proliferation, invasion, migration, capillary
Formation, differentiation and maturation.Vascular endothelial growth factor (VEGF) plays an important role in adjusting tumor vessel and occurring, wherein
VEGF-A represents most important angiogenesis stimulant.VEGF-A mainly by combined with VEGFR2 regulation and control HUVECs, activation
VEGFR2 can promote the phosphorylation of downstream PI3K-Akt, Erk1/2MAPK signal path, and PI3K-Akt, Erk1/2MAPK phosphoric acid
Change is closely related with proliferation, migration, the permeability of HUVECs, so as to promote angiogenesis together, so inhibiting any of the above described one
A process can inhibit angiogenesis.The HUVECs for VEGF-A how being inhibited to induce further to inquire into tilianin is proliferated, moves
The mechanism so as to inhibit inside and outside angiogenesis is moved, having studied tilianin by Western blot believes VEGF-A-VEGFR2
The influence of number access.
Method：By HUVEC cell inoculations in (2.3 × 10 in 6 orifice plates5/ hole), it is placed in 37 DEG C of incubators and is incubated for 24 hours.Using
After non-serum starved culture, add in (10-40 μM) of the tilianin incubation 2h of various concentration, and using be not added with the hole of drug as pair
According to then adding in 15ng/mL VEGF-A stimulates 15min.Buffering is cracked with the RIPA for adding phosphatase and protease inhibitors
Liquid lytic cell is to extract albumen, with BCA standard measure protein.The albumen of separation equivalent in PAGE gel, and with
200mA is transferred to pvdf membrane 90 minutes, and then film is incubated overnight with primary antibody so that antibody is combined with target protein at 4 DEG C.The
Two days recycling primary antibodies, film wash three times after (10 minute/time) with TBST buffer solutions, and the secondary antibody that film and HRP are conjugated is incubated at 4 DEG C
4 hours.After TBST is washed 3 times, make target protein band visible with ECL chemical illuminating reagents.
As a result with conclusion：The results are shown in Figure 5, and Fig. 5 is the VEGFR2 that tilianin inhibition is induced through VEGF-A and downstream letter
The schematic diagram of number access Akt, Erk1/2 phosphorylation；VEGF-A can induce VEGFR2 and its downstream signaling pathway Akt and Erk1/2
Phosphorylation, but the expression of these phosphorylated proteins is significantly lowered under the pretreatment of 20-40 μM of tilianin.
Basic principle, main feature and the advantages of the present invention of the present invention has been shown and described above.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, various changes and improvements may be made to the invention without departing from the spirit and scope of the present invention, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent defines.
Claims (10)
1. a kind of application of the tilianin and combinations thereof in anti-angiogenic medicaments are prepared.
2. application of the tilianin according to claim 1 and combinations thereof in anti-angiogenic medicaments are prepared, feature
It is：The anti-angiogenic medicaments are tilianin and combinations thereof.
3. application of the tilianin according to claim 2 and combinations thereof in anti-angiogenic medicaments are prepared, feature
It is：The angiogenesis is neonate tumour blood vessel, psoriatic lesions tissue blood vessel new life, Paget's diseases angiogenesis, good
The property angiogenesis of angiogenic diseases, the angiogenesis of arthritis pathological tissues, the blood vessel at atherosclerotic lesion are new
Raw or neovascular eye diseases angiogenesis.
4. application of the tilianin according to claim 3 and combinations thereof in anti-angiogenic medicaments are prepared, feature
It is：The neonate tumour blood vessel is angiogenesis caused by the angiogenesis or tumour of tumor tissues；
The neovascular eye diseases are primary or secondary neovascular eye diseases.
5. application of the tilianin according to claim 4 and combinations thereof in anti-angiogenic medicaments are prepared, feature
It is：The tumour is primary or secondary entity tumor.
6. application of the tilianin according to claim 1 and combinations thereof in anti-angiogenic medicaments are prepared, feature
It is：The dosage form of the drug is injection, capsule, tablet, powder, granule, pill, microcapsules and microsphere preparation, suppository, soft
Paste, spray or targeting preparation.
7. application of the tilianin according to claim 1 and combinations thereof in anti-angiogenic medicaments are prepared, feature
It is：The administering mode of the drug is oral, injection.
8. application of the tilianin according to claim 2 and combinations thereof in anti-angiogenic medicaments are prepared, feature
It is：The anti-angiogenic medicaments for tilianin and combinations thereof refer to tilianin single component as drug or tilianin and
Other pharmaceutically acceptable ingredients form composition.
9. application of the tilianin according to claim 8 and combinations thereof in anti-angiogenic medicaments are prepared, feature
It is：Other described pharmaceutically acceptable ingredients are one for not having the drug of antagonism with tilianin or pharmaceutically allowing
Kind or multiple auxiliary materials；
The tilianin is formed with other pharmaceutically acceptable ingredients in composition, and the weight content of active component tilianin is
0.1-99%.
10. according to claim 1 to 9 any one of them tilianin and combinations thereof answering in anti-angiogenic medicaments are prepared
With, it is characterised in that：The chemical structural formula of the tilianin is：
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111557947A (en) * | 2020-07-03 | 2020-08-21 | 新疆维吾尔自治区药物研究所 | Application of tilianin in preparation of medicine for improving expression of LXR-alpha |
| WO2021261707A1 (en) * | 2020-06-21 | 2021-12-30 | (주)앗코스텍 | Composition for alleviating, treating, or preventing muscular diseases, or improving muscular functions, containing korean mint extract or tilianin as active ingredient |
| CN114767699A (en) * | 2022-05-11 | 2022-07-22 | 新疆维吾尔自治区药物研究所 | Application of tilianin in preventive protection medicine for abdominal aortic aneurysm |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2021261707A1 (en) * | 2020-06-21 | 2021-12-30 | (주)앗코스텍 | Composition for alleviating, treating, or preventing muscular diseases, or improving muscular functions, containing korean mint extract or tilianin as active ingredient |
| CN111557947A (en) * | 2020-07-03 | 2020-08-21 | 新疆维吾尔自治区药物研究所 | Application of tilianin in preparation of medicine for improving expression of LXR-alpha |
| CN114767699A (en) * | 2022-05-11 | 2022-07-22 | 新疆维吾尔自治区药物研究所 | Application of tilianin in preventive protection medicine for abdominal aortic aneurysm |
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Application publication date: 20180608 |