CN106619612A - Application of Manoalide to preparation of medicament for inhibiting neovascularization - Google Patents
Application of Manoalide to preparation of medicament for inhibiting neovascularization Download PDFInfo
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- CN106619612A CN106619612A CN201610963323.XA CN201610963323A CN106619612A CN 106619612 A CN106619612 A CN 106619612A CN 201610963323 A CN201610963323 A CN 201610963323A CN 106619612 A CN106619612 A CN 106619612A
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- manoalide
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- 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
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses application of Manoalide to preparation of a medicament for inhibiting neovascularization. The fact that the Manoalide can inhibit the neovascularization is found for the first time, and the Manoalide can be applied to preparation of the medicament for inhibiting the neovascularization; the medicament can be widely applied to new vessel relevant diseases such as tumors, atherosclerosis, rheumatoid arthritis, regional ileitis, diabetic retinopathy, psoriasis, endometriosis and obesity, in particular to the neovascularization of the tumors. The Manoalide has important significance to clinical neovascularization-targeted tumor treatment.
Description
Technical field
The present invention relates to the new opplication field of known compound, in particular it relates to Manoalide is preparing the new green blood of suppression
Pipe forms the application in medicine.
Background technology
Angiogenesis refers to from the capillary for having existed the biological process for growing new blood vessel, and it is a series of
Very important link in physiology, pathologic process.Under physiological status, such as embryonic development, wound healing and in the menstrual cycle
Ovary, the cyclically-varying in uterus all rely on the blood vessel of new life and provide oxygen, nutriment and take away the waste of metabolism.Pathological state
Under Reperfu- sion after ischemic tissue, heart infarction, at this moment have obvious angiogenesis, this angiogenic process in the treatment should
It is promoted the treatment for being beneficial to disease.On the contrary, tumour, atherosclerotic, rheumatoid arthritis, segmental ileum
The diseases such as inflammation, BDR, psoriasis, endometriosis, obesity will be from angiogenesis inhibiting side
Face reaches the effect for the treatment of disease.
The medicine of current angiogenesis inhibiting has bevacizumab, rhEndostatin and reaction to stop, Sutent（Sunitinib）；Horse
Immediately take charge of him（Marimastat）；TNP-470 etc., but these medicines have respective limitation, such as hand-foot skin reaction, painstaking effort
The side effects such as pipe toxicity, renal toxicity, can increase the risk of lethal adverse events.Modal lethal adverse events are
Blood, myocardial infarction, hepatic failure, HP etc..
The content of the invention
Above-mentioned deficiency in order to overcome prior art of the invention, there is provided Manoalide is preparing suppression new vessels shape
Into the application in medicine.
To achieve these goals, the present invention is achieved by the following technical programs：
Claimed Manoalide is preparing the application in suppressing new vessels formation medicine.
Because chick chorioallantoic membrane（CAM）Model and chick embryo yolk sac film（YSM）Model is that the art is used to examine
Survey or screen and the typicalness with influence medicine and models with good availability are generated to new vessels, therefore, the present invention is only needed to
Prove that Manoalide can suppress new vessels to be formed in both representative models, it is possible to strong confirmation
Manoalide can be used for preparing the medicine for suppressing new vessels to be formed.
Preferably, claimed Manoalide is preparing the application in suppressing tumor angiogenesis medicine.
Preferably, claimed Manoalide is in suppression breast cancer tumour new vessels formation medicine is prepared
Application.
A kind of medicine for suppressing new vessels to be formed, containing Manoalide and pharmaceutically acceptable assistant agent.
Compared with prior art, the present invention has the advantages that：
Present invention firstly discovers that Manoalide can suppress in chick chorioallantoic membrane model the formation of two grades and three-level blood vessel,
And can suppress the growth of chick embryo yolk sac membrane modle medium vessels.In addition, planting breast cancer tumour on chick chorioallantoic membrane
Cell (MDA-MB-231), it is found that tumour cell grows and formed tumour on chick chorioallantoic membrane, and inside tumor occurs new
Angiogenic, after Manoalide process, this medicine can significantly inhibit the formation of inside tumor new vessels and the life of tumour cell
It is long, thus prove that Manoalide can suppress new vessels to be formed, can be used for preparing the medicine for suppressing new vessels to be formed, should
Medicine can be widely applied to new vessels relevant disease（As tumour, atherosclerotic, rheumatoid arthritis, segmental are returned
Enteritis, BDR, psoriasis, endometriosis, obesity etc.）.The especially angiogenesis of tumour,
Have great importance for the newborn treatment tumour of clinical target vascular therapy.
Description of the drawings
Fig. 1 is the observation result after drug-treated chick chorioallantoic membrane model.
Fig. 2 is the observation result after drug-treated chick embryo yolk sac membrane modle.
Fig. 3 is the observation result after drug-treated chick chorioallantoic membrane kind tumour cell.
Specific embodiment
The present invention is made with reference to Figure of description and specific embodiment further being elaborated, the embodiment
It is served only for explaining the present invention, is not intended to limit the scope of the present invention.Test method used in following embodiments is such as without spy
Different explanation, is conventional method；Material, reagent for being used etc., are the reagent for commercially obtaining if no special instructions
And material.
Embodiment 1
Chick chorioallantoic membrane（CAM）The foundation of model and drug-treated
1）Chicken embryo process：By 9 days chicken embryo surface cleaning of purchase, then wiped with 1: 1 000 bromogeramine liquid, wiped chicken after doing
Upwards, major axis and egg support are put into hatching in standard incubator to embryo air chamber in 45 °, design temperature (37.8 ± 0.5) DEG C and relatively wet
Degree 60%, daily according to egg turning egg(s) 3 times.
2）Windowing：It is incubated the 9th day, under candler air chamber is marked, it is determined that windowing position, after 75% alcohol disinfecting, with slowly
Fast dental burr bores a 1mm in air chamber mark3The aperture of size, with the curved tweezer of ophthalmology eggshell is carefully thrown off, and is formed diameter and is about
The breach of 1cm, with ophthalmic tweezers egg film exposure chick chorioallantoic membrane is gently taken off.
3）Experiment packet and dosing：Manoalide is obtained by buying, and chicken embryo is randomly divided into into two groups after windowing
（Manoalide experimental groups and DMSO control groups）.Per group of 8 chicken embryos.Experimental group adds medicine Manoalide to be measured, control group to add
Comparison liquid DMSO of same dose.Manoalide or DMSO are carefully added on chorioallantoic membrane from opening, medical proof fabric sealing,
37.8 DEG C are put into, are incubated 48 hours in the insulating box of 50%～60% humidity.
4）As a result observe：After dosing 48 hours, eggshell is cut off from center line, flow content in net egg, direct visual perception
The change of blood vessel on the chorioallantoic membrane of opening side, records phenomenon, and the photograph observation under Stereo microscope, counts vascular development feelings
Condition.
Observation index：Centered on so that point is administered, bending and attraction near referred to as blood vessel of the major blood vessel to spider；
The radial growth conditions towards center of peripheral vessels, referred to as blood vessel influx.Conversely, without this centripetal growing state, and than normal
Blood vessel reticular density under developmental state is little, and blood vessel is thin and quantity is substantially reduced, and the obvious avascular area domain of appearance then exists undetermined
Angiogenic growth development inhibitory action.
As a result as shown in figure 1, A attenuates for the blood vessel of the chick chorioallantoic membrane of Manoalide process, and two grades and three
Level vessel branch tails off.B is the statistics to vessel density growth rate, * * *P＜ 0.001.It can be seen that, Manoalide inhibits 9 days
The vascularization of chicken embryo CAM.
Embodiment 2
Chick embryo yolk sac film（YSM）The foundation of model and drug-treated
1）Hatching egg process：By hatching egg surface cleaning, then with 1:1 000 bromogeramine immersions steep 3min, wipe chicken embryo gas after doing
Upwards, major axis is put into hatching, design temperature (37.8 ± 0.5) DEG C and relative humidity in standard incubator with egg support in 45 ° for room
60%, daily according to egg, turning egg(s) 3 times.
2）Experimental technique：The hatching chicken embryo of 2.5 days is taken, ovum gallinaceum content is poured in sterilized petri dishes, will be red and black
Two an equal amount of falope rings of color marking pen mark are placed into the less diverse location of blood vessel on chick embryo yolk sac rete vasculosum,
Plate lid (being not required to closing) is covered, is put in 37～38 DEG C of incubators and is continued to be incubated 3～4h.After 3～4h, select falope ring and have no
Displacement, complete yolk cyst membrane, rete vasculosum and the well-developed chicken embryo of blastodisc, add medicine to be measured in the falope ring of red-label
Manoalide, the cushion rubber of density bullet adds comparison liquid DMSO of same dose as own control, then proceedes to incubation.
As a result observe：0h, 12h, 24h take pictures respectively under Stereo microscope after dosing, observe cushion rubber interior and surrounding
YSM vessel densities, trend.Shot in the whole silica gel ring in Experimental Area using the analysis of Image-ProPlus6.0 image analysis systems
Blood vessel, analysis indexes are turned to the change of vessel area and vessel density.
The observation result of 12h is as shown in Fig. 2 be the own control with 0.1%DMSO process on the left of A, right side is use after dosing
Result after Manoalide process；B is the statistics to vessel area growth rate.* P ＜ 0.05；* P ＜ 0.01.It can be seen that, DPPA
Inhibit the Angiogenesiss of 3 days chicken embryos YSM.
Embodiment 3
Chick chorioallantoic membrane（CAM）The foundation of inoculated tumour cell model and drug-treated：The chicken embryo hatching egg of purchase is used into 75%
Ethanol surface, upwards, major axis is put in standard incubator with egg support in 45 ° hatches chick embryo air sac, design temperature（38±
0.5）DEG C and relative humidity 60%, daily according to egg, turning egg(s), discard unfertilized or dead germ.8～10d is persistently incubated, air chamber is marked, really
Surely open a window position, with 75% alcohol disinfecting after, grind "+" vestige in mark with dental burr, then egg is carefully thrown off with the curved tweezer of ophthalmology
Shell, forms the breach of diameter about 1cm, and with cotton ball soaked in alcohol breach outer rim is wiped, and with the curved tweezer of ophthalmology egg film is gently taken off, exposes and contains
There is the chicken embryo yolk membrane tissue of abundant blood vessel, sealed in right-angled intersection with sterilizing medical proof fabric, adapt to 9～16 h.In super-clean bench
On open medical proof fabric.By aseptic cushion rubber be placed on vitellinae membrana first order vessel it is half side near.Sterilized silica gel medicine circle is put
In vitellinae membrana first order vessel and the intersection of two grades of blood vessels.Take the logarithm the MDA-MB-231 cells in growth period, with 0.25% pancreas
Enzymic digestion, PBS washings, 800rpm5min centrifugations go after supernatant to use 1mL PBS bases resuspended, and each hatching egg CAM is pressed after cell count
Silica gel medicine circle in add 50 μ L cell suspensions (containing about 3 × 106～5 × 106Individual cell), configure cell suspension and add egg
In, it is ensured that cushion rubber is close to CAM, to guarantee that the cell suspension for adding does not flow out cushion rubber.
After 30 h, adhesive plaster is opened on super-clean bench, observe tumour growth situation, eliminate no tumour growth on CAM films
Chicken embryo hatching egg.Chicken embryo hatching egg with tumour is randomly divided into into 4 groups, first 3 groups is experimental group, in cushion rubber plus 2 μM, 4 μM, 6 μ
A 50 μ L/Manoalide of M, control group gives the DMSO of same dose.Sealed with strile gauze dressing, incubator continues to be incubated.
24h dosings once, continuous dosing 3 times, it is ensured that reach the valid density of medicine.
About 16 h after 3rd dosing, along center line egg is cut off, and the content inside evacuation, PBS is stereoscopic micro-
Mirror is taken pictures and measures the volume of tumour, measures the area (V) of tumor region blood vessel, calculates MVD.Gross tumor volume=4 π/3（Major diameter ×
Minor axis2）.MVD (%)=tumor vessel area/(tumor region clear zone area+tumor region dark space area) × 100%.Will
CAM transplantable tumors take out, and put in PBS and clean.Embedding, section, H＆E dyeing.
H＆E is dyeed：H＆E dyeing is to take the paraffin section that the tumor tissues grown on chick chorioallantoic membrane are obtained, thickness
For 3～4 μm.Tissue paraffin section de-waxing, haematoxylin dyeing 4 minutes, anti-blue, eosin stains 3 seconds, 37 DEG C overnight, mounting.Just
Put each tissue blood vessel variable density of basis of microscopic observation and take pictures.
Claims (4)
1.Manoalide is preparing the application in suppressing new vessels formation medicine.
2.Manoalide is preparing the application in suppressing tumor angiogenesis medicine.
3.Manoalide is preparing the application in suppressing breast cancer tumour new vessels formation medicine.
4. a kind of medicine for suppressing new vessels to be formed, it is characterised in that containing Manoalide and pharmaceutically acceptable auxiliary
Agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610963323.XA CN106619612A (en) | 2016-10-28 | 2016-10-28 | Application of Manoalide to preparation of medicament for inhibiting neovascularization |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610963323.XA CN106619612A (en) | 2016-10-28 | 2016-10-28 | Application of Manoalide to preparation of medicament for inhibiting neovascularization |
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| CN106619612A true CN106619612A (en) | 2017-05-10 |
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| CN201610963323.XA Pending CN106619612A (en) | 2016-10-28 | 2016-10-28 | Application of Manoalide to preparation of medicament for inhibiting neovascularization |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4786651A (en) * | 1986-02-03 | 1988-11-22 | Allergan, Inc. | Treatment of cutaneous hyperproliferative dermatoses with manoalide |
| US4839385A (en) * | 1986-02-03 | 1989-06-13 | The Regents Of The University Of California | Use of manoalide and its derivatives for modifying calcium homeostasis |
| CN102532070B (en) * | 2011-12-29 | 2015-09-02 | 厦门大学 | One class is as the natural product and uses thereof of retinoid receptor (RARs) agonist |
-
2016
- 2016-10-28 CN CN201610963323.XA patent/CN106619612A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4786651A (en) * | 1986-02-03 | 1988-11-22 | Allergan, Inc. | Treatment of cutaneous hyperproliferative dermatoses with manoalide |
| US4839385A (en) * | 1986-02-03 | 1989-06-13 | The Regents Of The University Of California | Use of manoalide and its derivatives for modifying calcium homeostasis |
| CN102532070B (en) * | 2011-12-29 | 2015-09-02 | 厦门大学 | One class is as the natural product and uses thereof of retinoid receptor (RARs) agonist |
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