CN107496547A - One kind suppresses the garden burnet crude extract and preparation method of classical Wnt/β catenin paths - Google Patents
One kind suppresses the garden burnet crude extract and preparation method of classical Wnt/β catenin paths Download PDFInfo
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- CN107496547A CN107496547A CN201710873122.5A CN201710873122A CN107496547A CN 107496547 A CN107496547 A CN 107496547A CN 201710873122 A CN201710873122 A CN 201710873122A CN 107496547 A CN107496547 A CN 107496547A
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- 235000008291 Poterium sanguisorba Nutrition 0.000 title claims abstract description 42
- 244000173853 Sanguisorba officinalis Species 0.000 title claims abstract description 42
- 235000008282 Sanguisorba officinalis Nutrition 0.000 title claims abstract description 42
- 239000000287 crude extract Substances 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 108060000903 Beta-catenin Proteins 0.000 title claims abstract description 15
- 102000015735 Beta-catenin Human genes 0.000 title claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
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- 238000010828 elution Methods 0.000 claims abstract description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000002250 absorbent Substances 0.000 claims abstract description 5
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- 229940079593 drug Drugs 0.000 claims description 4
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- 230000002401 inhibitory effect Effects 0.000 abstract description 8
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- 238000003153 stable transfection Methods 0.000 abstract 1
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- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
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- 206010028980 Neoplasm Diseases 0.000 description 2
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- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- HQWTUOLCGKIECB-XZWHSSHBSA-N (6S,9aS)-6-[(4-hydroxyphenyl)methyl]-8-(1-naphthalenylmethyl)-4,7-dioxo-N-(phenylmethyl)-3,6,9,9a-tetrahydro-2H-pyrazino[1,2-a]pyrimidine-1-carboxamide Chemical compound C1=CC(O)=CC=C1C[C@H]1C(=O)N(CC=2C3=CC=CC=C3C=CC=2)C[C@H]2N1C(=O)CCN2C(=O)NCC1=CC=CC=C1 HQWTUOLCGKIECB-XZWHSSHBSA-N 0.000 description 1
- 102000016362 Catenins Human genes 0.000 description 1
- 108010067316 Catenins Proteins 0.000 description 1
- 241000050051 Chelone glabra Species 0.000 description 1
- 241001529246 Platymiscium Species 0.000 description 1
- 102100037664 Poly [ADP-ribose] polymerase tankyrase-1 Human genes 0.000 description 1
- 101710129670 Poly [ADP-ribose] polymerase tankyrase-1 Proteins 0.000 description 1
- 108700008625 Reporter Genes Proteins 0.000 description 1
- 240000006001 Thymus serpyllum Species 0.000 description 1
- KLGQSVMIPOVQAX-UHFFFAOYSA-N XAV939 Chemical compound N=1C=2CCSCC=2C(O)=NC=1C1=CC=C(C(F)(F)F)C=C1 KLGQSVMIPOVQAX-UHFFFAOYSA-N 0.000 description 1
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- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
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- HQVFCQRVQFYGRJ-UHFFFAOYSA-N formic acid;hydrate Chemical compound O.OC=O HQVFCQRVQFYGRJ-UHFFFAOYSA-N 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
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- 238000005259 measurement Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
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- 239000003960 organic solvent Substances 0.000 description 1
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- 239000000047 product Substances 0.000 description 1
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- 238000000926 separation method Methods 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 235000018553 tannin Nutrition 0.000 description 1
- 229920001864 tannin Polymers 0.000 description 1
- 239000001648 tannin Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000030968 tissue homeostasis Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/739—Sanguisorba (burnet)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of garden burnet crude extract and preparation method for suppressing classical Wnt/β catenin paths.The present invention relates to the preparation method for being used to suppress the garden burnet crude extract of Wnt paths, including pulverizing medicinal materials sieving, water to extract, the operations such as macroporous absorbent resin isolates and purifies.The garden burnet crude extract is through isolating and purifying, corresponding solution is made after freeze-drying, using construct stable transfection TCF response elements Wnt/ β catenin channel fluorescence element enzyme Reporter Systems 3T3 cells as experimental subjects, screening is tracked to the activity of crude extract.Crude extract of the present invention is 40% alcohol elution obtained through macroporous resin purification, and relative to water extract, active component is enriched with, notable to classical Wnt path inhibitory action, contributes to the quality control and subsequent product exploitation of garden burnet.
Description
Technical field
The present invention relates to the active component field of Chinese medicine, and in particular to Chinese medicine garden burnet crude extract suppress classical Wnt/ β-
The active component and preparation method of catenin paths.
Background technology
Garden burnet is one of China's traditional Chinese medicine, is used as medicine for rose family garden burnet platymiscium with dry root, rich in tannin, soap
The active skull cap components such as glycosides, flavonoids, in antimicrobial antiphlogistic, hemostasis antidiarrheal and anticancer etc. suffer from clear and definite drug effect and made
With, and there is good preventive and therapeutic effect for body leucocyte decline caused by carcinosis radiotherapy and chemotherapy, clinically obtain wide
General application.
Classical Wnt/ β-catenin paths are that biology is maintaining to grow, and tissue homeostasis etc. is vital
Signal path, the generation of its unconventionality expression and kinds cancer, development and invasion and attack transfer are closely related, therefore find the path
Effective inhibitor turn into one of effective means for the treatment of kinds cancer at present.It is from many document reports it can be found that existing more
Individual single compound plays inhibitory action in different phase to Wnt paths, as XAV939 is selected by suppressing tankyrase1/2
Property suppress the transcription of Wnt paths mediation, ICG-001 can antagonism Wnt/ β-catenin/TCF mediations transcription.And
Relative to micromolecular compound, Chinese medical extract has multicomponent feature, theoretically can be from different links, different targets
Point separately or concurrently acts on Wnt/ β-catenin paths, and dosage is less, and toxicity is lower, more advantageous.
The content of the invention
In view of the above-mentioned problems, the invention provides a kind of garden burnet crude extract for suppressing classical Wnt/β-catenin paths and
Preparation method.Extraction separation is carried out to garden burnet powder, these crude extracts carried out further according to luciferase reporter gene experiment
Screening active ingredients, the inactive material in crude extract is reduced in the case of retaining active ingredient as far as possible.
The technical solution adopted in the present invention is：
1. the preparation of garden burnet entirety crude extract：First medicine materical crude slice is dried to garden burnet to pulverize and sieve, entered with 8-12 times of solvent
Row refluxing extraction, reflux temperature is between 70 DEG C -90 DEG C, reflow's cycle 1-3 times, filters merging filtrate, and revolving recycling design is
.
2. garden burnet crosses the preparation at macroporous resin column difference graded ethanol elution position：Garden burnet dried powder is dissolved with suitable quantity of water
Afterwards, it is splined on D101 macroporous absorbent resins, and different proportion alcohol-water volume ratio (0: 100, (40-50): (60-50), 90: 10)
3-4 column volume is eluted successively, is collected eluent recovery organic solvent, is freeze-dried and produces.
3. each crude extract of garden burnet suppresses the Activity determination of Wnt/ β-catenin paths：Each crude extract is made into one with ultra-pure water
Determine the solution of concentration, the filtering with microporous membrane with 0.22 μm is degerming in an aseptic environment, will be thin using 3T3 cells as experimental subjects
Born of the same parents are inoculated in the adherent 24h of 96 orifice plates, and different pharmaceutical is handled into cell 24h with series concentration, and medicine is detected to cell using mtt assay
Toxic action.
In the range of 20% cytotoxic concentration, drug-treated cell 24h, examined using Steady-Glo@Assay reagents
Survey the intensity of variation of Wnt paths.
The beneficial effects of the invention are as follows：Searched out in the inventive method has in strong inhibition effect to Wnt paths
Medicine Radix Sangusorbae extract, and more than 50% active substance is eliminated using macroporous absorbent resin during isolating and purifying, have
Beneficial to the quality control of Chinese medical extract.
Brief description of the drawings
Fig. 1 is：Garden burnet entirety water extract to 3T3 cytotoxic effects (Figure 1A), and wnt3a stimulants effect under
Inhibitory activity (Figure 1B) of the elm entirety water extract to Wnt paths.
Fig. 2 is：Garden burnet crosses macroporous resin column alcohol-water volume ratio 0/100 and elutes position to 3T3 cytotoxic effects (figure
2C), to the inhibitory activity (Fig. 2 D) of Wnt paths and under the effect of wnt3a stimulants.
Fig. 3 is：Garden burnet crosses macroporous resin column alcohol-water volume ratio 40/60 and elutes position to 3T3 cytotoxic effects (figure
3E), to the inhibitory activity (Fig. 3 F) of Wnt paths and under the effect of wnt3a stimulants.
Fig. 4 is：Garden burnet crosses macroporous resin column alcohol-water volume ratio 90/10 and elutes position to 3T3 cytotoxic effects (figure
4G), to the inhibitory activity (Fig. 4 H) of Wnt paths and under the effect of wnt3a stimulants.
Fig. 5 is：Garden burnet crosses the elution of macroporous resin column alcohol-water volume ratio 40/60 position and prepares gained sample in triplicate
HPLC chromatogram.
Embodiment
Embodiment 1
1. prepared by garden burnet entirety water extract：
Garden burnet dried powder (crossing 60 mesh sieves) 30g is taken to add 300ml 80 DEG C of distilled water water-bath to be heated to reflux 3 times, each 1h,
Merging filtrate is filtered, rotates to paste and calculating yield 41.4% is collected after being freeze-dried.
2. garden burnet crosses the preparation at macroporous resin column ethanol gradient elution position：
Garden burnet entirety water extract 2.0g is taken to be used with D101 macroporous absorbent resins are splined on after distilled water ultrasonic dissolution 30min
The ratio that alcohol-water volume ratio is 0/100,40/60,90/10 elutes 4 column volumes successively, after gained eluent reclaims ethanol
Freeze-drying, volume ratio is 40% alcohol elution yield 48.46%.
3. garden burnet crosses the HPLC analyses that macroporous resin column alcohol-water volume ratio 40/60 elutes position：
It is 40% ethanol elution to prepare garden burnet to cross macropore resin column product ratio according to the 2nd step experimental method 3 repeated experiments
Position yield is respectively 45.56%, 48.46%, 45.17%.It is made into 10mg/ml solution with ultra-pure water, 10ul sampling volumes,
HPLC is detected.
Liquid-phase condition：A：0.2% formic acid water B：Methanol
Elution program：0-2min (2%B)；2-5min (5%B)；5-10min (12%B)；10-15min (20%B)；15-
35min (30%B)；35-50min (45%B)；50-60min (60%B)；60-70min (70%B)；70-75min (80%
B)；75-80min (40%B)；80-85min (5%B)
4. Drug inhibition Wnt pathway activities detect：
A. cell culture：3T3 cells, cultivated in containing 10% serum DMEM culture mediums, and it is dense to be positioned over carbonated
Degree 5%, temperature are in 37 DEG C of incubator.
B. passage：When cell density length to 70%-80%, culture medium is sucked, is washed 2 times with sterile PBS, is used
0.25% pancreatin digestion 1min or so, adds 2ml serum-containing media and terminates digestion, centrifuge 3min under 1000rpm, discard
Culture medium, continue to cultivate in 10% serum DMEM culture mediums.
C. plating cells：Cell is resuspended after pancreatin digests with culture medium, with cell counting count board meter after dilution certain multiple
Number, by 2x104Individual/hole density is inoculated in 96 orifice plates, and 100ul cell suspension is added per hole.
D. cell administration：Adherent 24h after cell inoculation, the garden burnet crude extract decoction for adding setting concentration continue to cultivate 24h.
E. cytotoxicity detects：Culture medium is discarded after cell administration 24h, adds 20ul 5mg/ per hole under the conditions of lucifuge
Ml MTT decoctions 4h in incubator, the DMSO for sucking decoction addition 150ul per hole is taken out after 10min is shaken on shaking table,
Absorbance is detected under 570nm wavelength, calculates cell survival rate.
F. the detection of Wnt pathway activities is suppressed：Abandoned after cell administration 24h under the conditions of the wnt3a stimulants containing 100ng/ml
Remove culture medium, washed 1 time with PBS, the Steady-Glo@Assay reagents that mix in advance of 30ul are added per hole, at room temperature in
10min is shaken on shaking table, by Steady-Glo@Assay agent transfers into blank, in multi-function microplate reader
The activity of luciferase is determined under luminescence measurement functions.
5. data analysis：
The data obtained imports GraphPad Prism5 and analyzed.
6. experimental result：
A. garden burnet entirety water extract has obvious inhibition (figure in the range of 20% cytotoxicity to Wnt paths
1)。
B. garden burnet separates through large pore resin absorption column, and pure water elution position does not have inhibitory activity (Fig. 2) to Wnt paths.
C. garden burnet separates through large pore resin absorption column, the suppression of the elution position of alcohol-water volume ratio 40/60 to Wnt paths
Make active (Fig. 3) suitable with overall water extract.
D. garden burnet separates through large pore resin absorption column, and the elution position of alcohol-water volume ratio 90/10 does not have to Wnt paths
Obvious inhibitory activity (Fig. 4).
E. garden burnet separates through large pore resin absorption column, the chromatograms explanation at the elution position of alcohol-water volume ratio 40/60
The site component obtained by the preparation method that the present invention uses is more or less the same, and has repeated (Fig. 5).
Claims (6)
1. one kind suppresses the garden burnet crude extract and preparation method of classical Wnt/β-catenin paths, it is characterised in that：Garden burnet slightly carries
Thing includes the garden burnet prepared slices of Chinese crude drugs after pulverizing and sieving, and is extracted with the solvent of certain volume, and isolated and purified through macroreticular resin
Sample afterwards.
2. a kind of garden burnet crude extract and preparation method for suppressing classical Wnt/β-catenin paths according to claim 1,
It is characterized in that：After garden burnet medicine materical crude slice pulverizes and sieves, to carry out refluxing extraction relative to the solvent of 8-12 times of the multiple of quality grams.
3. a kind of garden burnet crude extract and preparation method for suppressing classical Wnt/β-catenin paths according to claim 2,
It is characterized in that：The solvent of refluxing extraction is distilled water, and for refluxing extraction temperature at 70 DEG C -90 DEG C, reflow's cycle is 1-3 times, should
Step extraction yield control is in 40%-50%.
4. a kind of garden burnet crude extract and preparation method for suppressing classical Wnt/β-catenin paths according to claim 1,
It is characterized in that：When macroporous absorbent resin further isolates and purifies to garden burnet crude extract, macroreticular resin model D101, elute molten
Agent is the alcohol-water of different proportion.
5. a kind of garden burnet crude extract and preparation method for suppressing classical Wnt/β-catenin paths according to claim 4,
It is characterized in that：The volume ratio of the ratio ethanol/water of eluting solvent is 0/100 during macroreticular resin elution samples, (40-50)/
(60-50), 90/10, gradient elution, the elution volume of each gradient is 3-5 column volume.
6. a kind of garden burnet crude extract and preparation method for suppressing classical Wnt/β-catenin paths according to claim 5,
It is characterized in that：Each position yield obtained during macroreticular resin gradient elution is controlled in 40%- for the yield of 0/100 gradient
The yield control of 45%, (40-50)/(60-50) gradient is controlled in 2%-5% in 45%-49%, the yield of 90/10 gradient.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110882315A (en) * | 2019-12-18 | 2020-03-17 | 哈尔滨医科大学 | Sanguisorba officinalis extract for preventing colon cancer and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030044478A1 (en) * | 2001-08-15 | 2003-03-06 | Epstein Howard A. | Burnet extract |
| CN101862385A (en) * | 2009-04-20 | 2010-10-20 | 成都地奥制药集团有限公司 | Sanguisorba saponins and preparation method of sanguisorbin I |
-
2017
- 2017-09-15 CN CN201710873122.5A patent/CN107496547A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030044478A1 (en) * | 2001-08-15 | 2003-03-06 | Epstein Howard A. | Burnet extract |
| CN101862385A (en) * | 2009-04-20 | 2010-10-20 | 成都地奥制药集团有限公司 | Sanguisorba saponins and preparation method of sanguisorbin I |
Non-Patent Citations (1)
| Title |
|---|
| 王振飞等: "地榆水提液对四种癌细胞生长抑制作用的研究", 《时珍国医国药》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110882315A (en) * | 2019-12-18 | 2020-03-17 | 哈尔滨医科大学 | Sanguisorba officinalis extract for preventing colon cancer and preparation method and application thereof |
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Application publication date: 20171222 |