CN104548067A - Application of recombinant human endostatin in preparing drugs for treating ocular neovascular diseases - Google Patents
Application of recombinant human endostatin in preparing drugs for treating ocular neovascular diseases Download PDFInfo
- Publication number
- CN104548067A CN104548067A CN201410814133.2A CN201410814133A CN104548067A CN 104548067 A CN104548067 A CN 104548067A CN 201410814133 A CN201410814133 A CN 201410814133A CN 104548067 A CN104548067 A CN 104548067A
- Authority
- CN
- China
- Prior art keywords
- recombinant human
- vascular endothelial
- group
- corneal
- neovascular diseases
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses the application of recombinant human endostatin in preparing drugs for treating ocular neovascular diseases. Researches show that the recombinant human endostatin can remarkably inhibit the formation of ocular neovasculars, is safe, non-toxic and free of drug resistance, and has wide application prospects in preparing drugs for treating ocular neovascular diseases.
Description
Technical field:
The invention belongs to biological pharmacy technical field, be specifically related to the application of recombinant human vascular endothelial inhibin in preparation treatment eye table neovascular diseases medicine.
Technical background:
The common name of rhEndostatin (Endostar) is recombinant human vascular endothelial inhibin injection, it is a kind of novel recombinant human blood vessel endothelium suppression medicine of Chinese scholar independent research, Endostar produces with recombinant DNA technology, adopts the recombinant human vascular endothelial inhibin that escherichia coli are produced as protein expression system.The mechanism of action of rhEndostatin is that Tumor suppression new vessels is formed, and blocks the nutrition supply of tumor cell, finally reaches the object of " dying of hunger " tumor cell.RhEndostatin is first class national new drug, does not belong to chemotherapeutics, and tumor grows in vivo and reaches 2mm
3time above, the blood vessel network conveying nutrient substance of self must be formed, otherwise can not continued growth.Tumor suppression vascularization, cuts off its nutrition supply, and after a period of time, tumor will be in resting state, and the result of long term causes death of neoplastic cells, and this Therapeutic Method is called tumor starvation.The medicine of all Antineoplastic angiogenesis all belongs to this therapy.
Ocular angiogenesis is eye due to tissue ischemia, anoxia and cause new vessels to be formed as the disease of pathological change.Due to the exception of new vessels morphology and function, its with occur ooze out, breed, the pathological changes such as hemorrhage, the destruction of normal eye 26S Proteasome Structure and Function can be caused, cause serious visual function damage.Wherein eye table neovascular diseases can betide corneal chemical burn, lacerated wound, infective inflammation, immunity oculopathy, exposure keratitis, pterygium, trichiasis, corneal degeneration, malnutrition and under previously the operation of eye table comprises the multiple pathologic conditions such as keratoplasty, is one of common diseases causing blindness.Its concrete pathogenesis is still not clear, chief reason is that between ophthalmic angiogenic factors (VEGF, fibroblast growth factor, transforminggrowthfactor-β1, interleukin 1 etc.) and angiostatin (matrix metalloprotease, nitric oxide synthase, endogenous endothelial inhibin, heparin sulfate etc.), regulatory mechanism is unbalance, relevant with corneal edema, inflammation, anoxia etc.
The treatment of eye table new vessels is a great problem, does not also have the Therapeutic Method of specific medicament and specially good effect so far.The main method applied clinically at present has laser photocoagulation, photodynamic therapy, hands art; Conventional medicine is mainly hormone, vegf antagonist, Chinese patent medicine etc.Though these treatments have certain curative effect, also there is the drawback of damaging normal tissue, offer limited effectiveness.Laser, operation all can only be carried out at subrange, and the prolonged application of hormone can bring a series of side reaction, and photodynamic therapy, Angiogenesis antagonist is expensive and often need repeatedly treat, and cannot reverse the pathological changes produced, and can not produce eyesight improving at a specified future date.Eye table new vessels constitutes a serious threat to patient's vision, needs more effective Therapeutic Method and medicine.
Summary of the invention:
The object of the present invention is to provide the application of recombinant human vascular endothelial inhibin in preparation treatment eye table neovascular diseases medicine.
Object of the present invention is achieved through the following technical solutions:
The application of recombinant human vascular endothelial inhibin in preparation treatment eye table neovascular diseases medicine.
Described eye table neovascular diseases is that eye table a variety of causes causes new vessels to invade transparency cornea, comprises eye table chemical injury, corneal penetrating injury, infected keratitis, thermal burn, trachoma, epidemic keratoconjunctivitis, exposure keratitis, immunity oculopathy, Terrien marginal degeneration, corneal rupture wound, corneal degeneration, cerneal dystrophy, long periods of wear contact lens, the cornea rebirth blood vessel of corneal graft and previously other operation on cornea generation, pterygium, keratoconus, cornea Fructus Vitis viniferae kind, trichiasis and Stevens-Johnson syndrome; Or be retinal neovascularization etc.Especially eye table chemical injury, corneal penetrating injury, infected keratitis, operation on cornea formed new vessels, pterygium and Stevens-Johnson syndrome.
Described medicine comprises the adjuvant of recombinant human vascular endothelial inhibin and pharmaceutically acceptable carrier or conventional use.
The dosage form of described medicine is ejection preparation.
Beneficial effect of the present invention:
Research shows, recombinant human vascular endothelial inhibin can suppress the formation of eye table new vessels significantly, and safety non-toxic, have no drug resistance, show anti-neovascularization medicaments at treatment eye and have broad application prospects.
Detailed description of the invention:
In order to better the present invention is described, below in conjunction with drug study, zoopery and clinical experiment, its purposes prepared eye and show in anti-neovascularization medicaments is described, if no special instructions, hereinafter involved experiment all obtains by commercial sources with material.
1, recombinant human vascular endothelial inhibin injection (rhEndostatin) is on the impact of vascular endothelial cell proliferation
1.1 experiment material
Cell: human umbilical vein endothelial ECV304 is provided by Chinese Academy of Sciences's Shanghai cell bank.
Main biochemical reagent: recombinant human vascular endothelial inhibin injection (rhEndostatin), Shandong Xiansheng Maidejin Biological Pharmaceutical Co., Ltd. produces
1.2 experimental technique
Carried out recovering, cultivate, go down to posterity and identifying by human umbilical vein endothelial ECV304, trophophase cell of taking the logarithm, is diluted to 5 × 10 with PBS buffer
3/ mL density is inoculated in 96 well culture plates, every hole 100 μ L, respectively to the recombinant human vascular endothelial inhibin solution 100 μ L/ hole adding the different dilution factors (5000 μ g/mL, 2500 μ g/mL, 1250 μ g/mL, 625 μ g/m, 313 μ g/mL, 125 μ g/mL) that PBS dilutes in hole, often organize and all establish 6 multiple holes, matched group does not add recombinant human vascular endothelial inhibin, and adding equal volume has serum free culture system liquid.In 37 DEG C, 5%CO
2cultivating in incubator after 24h, 48h, 72h days uses MTT (tetramethyl azo azoles salt) method to dye, and at microplate reader 490nm place determination experiment group and matched group absorbance, and calculates cell proliferation inhibition rate, the results are shown in Table 1.
Table 1 recombinant human vascular endothelial inhibin is to the cell proliferation inhibition rate of ECV304
Result shows, observe under difference inverted microscope recombinant human vascular endothelial inhibin effect 24,48, the form of ECV304 living cells, without significant change, has no cell rounding and cell and takes off wall and suspend after 72h.Cell number comparatively matched group minimizing in rarely seen visual field, propagation slows down.To the Inhibit proliferaton effect of the ECV304 cell cultivated as can be seen from Table 1, suppression ratio is concentration dependent to different concentration, the highest can nearly 54%.
2, retinal neovascularization mouse model experiment
2.1 materials and methods
Laboratory animal: the healthy C57BL/6J neonatal rat 80 (80 eyes) of Mus 7d in age, weight is about 30g, and male and female are not limit, and jointly raises with the female Mus of suckling.Mice and female Mus are purchased from Beijing HFK Bio-Technology Co., Ltd..
Main agents: rhEndostatin (Shandong Xiansheng Maidejin Biological Pharmaceutical Co., Ltd., 3mL:15mg).
80 mices are divided into Normal group, hyperoxia matched group, saline control group and rhEndostatin intervention group at random, often organize 20.Normal group is raised under being placed in normal air environment, and excess-three group is placed in airtight in oxygen case after 5d, and Normal group and hyperoxia matched group do not deal with, saline control group intraperitoneal injection of saline, rhEndostatin intervention group lumbar injection rhEndostatin.
The induced retinal neovascularization models in mice of mice is set up in 2.2 hyperoxia inductions
Be positioned in airtight plexiglass box together with female to Mus 7d in age mice and suckling Mus, add a cover with bigger glass above glass box, access volume fraction 100% medical pure oxygen in container, oxygen flux control is 0.50 ~ 0.75L/min, oxygen concentration in container is made to remain (75 ± 2) %, first oxygen flow is heightened to 5L/min and be about 3min, after making it permit rapidly full oxygen case, turn down again to partial pressure of oxygen and balance, about 0.7L/min, and with the oxygen concentration in oxygen analyser monitored over time container, guarantee that oxygen concentration is constant.Open oxygen case every day to change bedding and padding, add food, change water, replace female Mus 1 time.Raise 5d under oxygen environment, then place in Normal Environmental Temperature and raise.Ambient temperature controls as (23 ± 2) DEG C.
After modeling success (Mus 12d in age), according to Mice Body quality, draw the rhEndostatin diluent or normal saline that prepare according to 25 μ L/g, along hypogastric region inserting needle in mice, inject after pumpback depletion of blood, rhEndostatin diluent concentration is 30 μ L stock solution/1mL normal saline.Injection 1 time/d, injects 5 times altogether, and 5d post processing is drawn materials observation.
After administration, the 5th day (Mus 17d in age) by the intraperitoneal anesthesia of each group of mice row till death, then wins eyeball, is placed in formalin and fixes 24h, dehydration, paraffin embedding.Avoid when getting section around optic disc, the sagittal plain along eyeball does retina serial section, and slice thickness is 5 μm, 30 μm, interval, and every eyeball chooses 4 sections, and conventional dewaxing is dyeed through HE.Often open section random selecting 3 high power fields (× 400), count the nucleus of vascular endothelial cells number of the breakthrough inner limiting membrane contained by each visual field with double-blind method, in vitreous chamber except other and the unrelated nucleus of vascular endothelial cells of internal limiting membrane.To break through the index of nucleus of vascular endothelial cells as counting retinal neovascularization of inner limiting membrane.
Four groups of retinal neovascularization counting comparative results display: Normal group retinal neovascularization is less, the only visible nucleus of vascular endothelial cells breaking through inner limiting membrane in minority section.Hyperoxia matched group nucleus of vascular endothelial cells number mouthful increases obviously.The visible more nucleus of vascular endothelial cells of saline control group.RhEndostatin intervention group nucleus of vascular endothelial cells number reduces.Normal group, hyperoxia matched group, saline control group and rhEndostatin intervention group nucleus of vascular endothelial cells count be respectively (0.85 ± 0.12), (25.65 ± 1.06), (23.32 ± 1.32), (2.95 ± 0.35) individual/HP; Normal group and hyperoxia matched group comparing difference have statistical significance (P<0.01), induced retinal neovascularization models in mice success.Hyperoxia matched group and saline control group break through the neovascular endothelium cell check figure mouth of internal limiting membrane obviously more than Normal group and rhEndostatin intervention group (P<0.01), and rhEndostatin intervention group is more than Normal group (P<0.01).
Conclusion shows: recombinant human vascular endothelial inhibin can suppress the formation of retinal neovascularization significantly, can reach the effect of effectively treatment retina neovascular diseases.
3, recombinant human vascular endothelial inhibin suppresses the test of cornea rebirth blood vessel after rabbit suture
3.1 materials and methods
Laboratory animal: new zealand white rabbit 24, body weight 2.0-2.5kg, male and female have concurrently, and by Nanchang, Long Pingtu industry company limited provides.
Main agents: rhEndostatin (recombinant human vascular endothelial inhibin injection), Yantai Mai get Jin biological engineering limited company produces, and 15mg/ props up (3mL).Dexamethasone 5mg/ props up, and Feng Yuan pharmaceutical factory of Ma'an Mountain produces.
Prepared by rabbit corneal neovascularization model: rabbit gives ketalar (50mg/kg) and chlorpromazine (10mg/kg) intramuscular injection anesthesia.All getting right eye is art eye, routine disinfection, rinses conjunctival sac, Oxybuprocaine hydrochloride eye drops topical anesthesia 3 times.Eyelid left by eye speculum, adopts cutting needle (3/8) to wear 5-0 silk thread, does 3 pin sutures above cornea, suture upper end distance limbus of corneae 2.5mm, s degree of depth 1/2-1/3 corneal thickness, suture is imbedded corneal stroma and is about 3mm, stay the end of a thread to be about 1mm at anterior corneal surface, pin is avoided entering anterior chamber as far as possible.
Grouping and process: experimental rabbit is divided into 3 groups by random digits table, often organize 8.A group is blank group, and suture plays sub-conjunctival injection normal saline 0.1mL in postoperative 1st day.The next day 1 time.B group is experimental group, within postoperative 1st day, plays sub-conjunctival injection rhEndostatin 0.1mL, the next day 1 time.C group is matched group, within postoperative 1st day, plays sub-conjunctival injection dexamethasone 0.1ml, the next day 1 time.
3.2 result
There is ciliary congestion in the 2nd day after suture in each group of cornea, the 3rd day visible limbus of corneae is that hairbrush shape new vessels grows to suture place, and within the 7th day, can reach suture place, within the 13rd day, new vessels area reaches peak, and the rear speed of growth is slack-off.Cornea rebirth blood vessel growing state: A group neovascularization growth is the most vigorous, blood vessel is intensive, thick, and slowly, blood vessel is tiny, sparse for B group and C group neovascularization growth.B group, C group each time point new vessels length, area after corneal suture are all less than A group, and difference has statistical significance (P<0.001).B group is slightly strong compared with the inhibitory action of C group corneal new vessels, has statistical significance (P<0.05) the 13rd day and the 18th day difference between the two.In table 2.
Table 2 cornea rebirth blood vessel length and Area comparison
4. recombinant human vascular endothelial inhibin hinders the impact of the cornea rebirth blood vessel caused on rat chemical
SD rat 20, body weight 160 ~ 200g, after adopting silver stick to cause cornea of rats chemical burn, divide two groups, wherein one group according to Mice Body quality, draws the rhEndostatin diluent or normal saline that prepare, along hypogastric region inserting needle in mice according to 25 μ L/g, inject after pumpback depletion of blood, rhEndostatin diluent concentration is 30 μ L stock solution/1mL normal saline.Injection 1 time/d, injects 5 times altogether, and 5d post processing is drawn materials observation.Another group according to 25 μ L/g injecting normal salines in contrast.10 rat corneal neovascularization of observed result display injecting normal saline all grow vigorous, and blood vessel is intensive, thick; 10 rat freshman angiogenic growths of injection rhEndostatin diluent are slow, and blood vessel is tiny, sparse.Result display rhEndostatin can significantly suppress rat chemical to hinder the cornea rebirth blood vessel caused.
5. the clinical therapeutic effect to recurrent pterygium of recombinant human vascular endothelial inhibin
For 15 routine recurrent pterygiums, simple except after art excision, positive common medicine antiinflammatory, 5 days after operation is taken out stitches; Material is controlled: to the left eye intravitreal injection rhEndostatin injection 0.2ml of patient after taking out stitches, injection in every 2 weeks once, it within 2 months, is a course for the treatment of, all disappear from the crawl toward shallow-layer new vessels of limbus of corneae of conjunctiva in 15 routine patients after medication, follow up a case by regular visits to and have no recurrence in 9 months, after medication, have no irritation and systemic side effects.
6. conclusion
The cornea rebirth blood vessel propagation that rhEndostatin can effectively suppress chemical injury, operation on cornea causes, and recurrent pterygium curative effect is better than the method for current clinical conventional hormone therapy eye table new vessels.
Claims (4)
1. the application of recombinant human vascular endothelial inhibin in preparation treatment eye table neovascular diseases medicine.
2. application according to claim 1, it is characterized in that described eye table neovascular diseases for shown chemical injury, corneal penetrating injury, infected keratitis, thermal burn, trachoma, epidemic keratoconjunctivitis, exposure keratitis, immunity oculopathy, Terrien marginal degeneration, corneal rupture wound, corneal degeneration, cerneal dystrophy, long periods of wear contact lens and operation on cornea by eye and formed cornea rebirth blood vessel, pterygium, keratoconus, cornea Fructus Vitis viniferae kind, trichiasis and Stevens-Johnson syndrome; Or be retinal neovascularization.
3. application according to claim 1, is characterized in that described medicine comprises the adjuvant of recombinant human vascular endothelial inhibin and pharmaceutically acceptable carrier or conventional use.
4. application according to claim 1, is characterized in that the dosage form of described medicine is ejection preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410814133.2A CN104548067A (en) | 2014-12-23 | 2014-12-23 | Application of recombinant human endostatin in preparing drugs for treating ocular neovascular diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410814133.2A CN104548067A (en) | 2014-12-23 | 2014-12-23 | Application of recombinant human endostatin in preparing drugs for treating ocular neovascular diseases |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104548067A true CN104548067A (en) | 2015-04-29 |
Family
ID=53065836
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410814133.2A Pending CN104548067A (en) | 2014-12-23 | 2014-12-23 | Application of recombinant human endostatin in preparing drugs for treating ocular neovascular diseases |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104548067A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112121149A (en) * | 2020-09-30 | 2020-12-25 | 中南大学湘雅三医院 | Medicine for treating postoperative abdominal adhesion and new application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002067971A2 (en) * | 2001-02-22 | 2002-09-06 | Novartis Ag | Use of endostatin in the treatment of ocular neovascularization |
| WO2006119128A2 (en) * | 2005-04-29 | 2006-11-09 | Research Development Foundation | Vascular targeting of ocular neovascularization |
| CN101219206A (en) * | 2008-02-01 | 2008-07-16 | 山东先声麦得津生物制药有限公司 | Application of recombinant human vascular endothelial inhibin in pharmacy |
| CN101224296A (en) * | 2008-02-01 | 2008-07-23 | 山东先声麦得津生物制药有限公司 | Stable recombinant human endostatin preparation and preparation process thereof |
| CN102233132A (en) * | 2010-04-28 | 2011-11-09 | 成都康弘生物科技有限公司 | Application of VEGF acceptor fusion proteins in preparation of drugs for inhibiting growth of ocular surface neovascularization |
-
2014
- 2014-12-23 CN CN201410814133.2A patent/CN104548067A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002067971A2 (en) * | 2001-02-22 | 2002-09-06 | Novartis Ag | Use of endostatin in the treatment of ocular neovascularization |
| WO2006119128A2 (en) * | 2005-04-29 | 2006-11-09 | Research Development Foundation | Vascular targeting of ocular neovascularization |
| CN101219206A (en) * | 2008-02-01 | 2008-07-16 | 山东先声麦得津生物制药有限公司 | Application of recombinant human vascular endothelial inhibin in pharmacy |
| CN101224296A (en) * | 2008-02-01 | 2008-07-23 | 山东先声麦得津生物制药有限公司 | Stable recombinant human endostatin preparation and preparation process thereof |
| CN102233132A (en) * | 2010-04-28 | 2011-11-09 | 成都康弘生物科技有限公司 | Application of VEGF acceptor fusion proteins in preparation of drugs for inhibiting growth of ocular surface neovascularization |
Non-Patent Citations (5)
| Title |
|---|
| 宋修军等: "《临床骨科药物学》", 31 May 2010, 科学技术文献出版社 * |
| 张平等: "转内皮抑制素基因治疗大鼠角膜新生血管", 《国际眼科杂志》 * |
| 李彩云等: "重组人血管内皮抑素对视网膜新生血管的抑制作用", 《中华实用诊断与治疗杂志》 * |
| 李维义等: "重组人内皮抑素两种给药途径抑制角膜新生血管的实验研究", 《农垦医学》 * |
| 钟敏等: "重组人血管内皮抑制素抑制角膜新生血管的实验研究", 《南昌大学学报（医学版）》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112121149A (en) * | 2020-09-30 | 2020-12-25 | 中南大学湘雅三医院 | Medicine for treating postoperative abdominal adhesion and new application thereof |
| CN112121149B (en) * | 2020-09-30 | 2023-10-13 | 中南大学湘雅三医院 | Medicine for treating postoperative abdominal adhesion and new application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104490861A (en) | Sustained-release nepafenac eye-drops preparation | |
| WO2015135306A1 (en) | Uses of artemisinin and derivatives thereof in manufacture of medicaments for prevention and treatment of vascular diseases in ophthalmology and pharmaceutical compositions | |
| CN102058581B (en) | Ophthalmic gel | |
| JP2018527147A (en) | Method for producing a chronic ocular hypertension animal model | |
| Stanzel et al. | Surgical approaches for cell therapeutics delivery to the retinal pigment epithelium and retina | |
| CN102166205B (en) | The medical usage of paeonol and derivant thereof | |
| CN104548067A (en) | Application of recombinant human endostatin in preparing drugs for treating ocular neovascular diseases | |
| CN102258518A (en) | Application of huperzine A to preparation of medicament for treating glaucoma and/or ischemia-induced optic nerve damage | |
| CN101797223A (en) | Huperzine A preparations for eyes and application thereof | |
| FINE et al. | Therapy of experimental intraocular Aspergillus infection | |
| CN105214071A (en) | A kind of size variance eye drop and preparation method thereof | |
| Liou et al. | Effects of intraocular epinephrine on the corneal endothelium of rabbits | |
| WO2015081854A1 (en) | Tribulus terrestris saponin intravitreal injection administration system and uses thereof | |
| CN113577066B (en) | Use of arylguanidine compounds or pharmaceutically acceptable salts thereof | |
| WO2015081850A1 (en) | Novel intravitreal injection drug delivery system for mouse nerve growth factor, and application thereof | |
| Shields et al. | Neovascular glaucoma associated with an iris melanoma: a clinicopathologic report | |
| CN105012305A (en) | Application of lamivudine and pharmaceutical salt thereof in treating age-related macular degeneration | |
| CN110200904A (en) | A kind of drop intraocular pressure sustained release eye drop composition and preparation method thereof | |
| RU2558991C1 (en) | Method for simulating proliferative retinopathy in rats | |
| CN103169717A (en) | Application of anthracene nucleus antibiotic and its pharmaceutical salt for treating ocular inflammation | |
| CN102349901A (en) | Application of pirfenidone in preparation of medicaments for controlling proliferative diseases after ophthalmologic operation and eye drops thereof | |
| CN109776656B (en) | Polypeptide TIN7N for inhibiting angiogenesis and application thereof | |
| CN118726349A (en) | RNA molecule for treating fundus vascular diseases and application thereof | |
| RU2303457C1 (en) | Method for applying enzymotherapy | |
| Gawshinde et al. | A REVIEW ON OCUULAR DRUG DELIVERY |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150429 |