CN108047037A - A kind of correspondence-kauran diterpene derivative and its application in antitumor drug is prepared - Google Patents
A kind of correspondence-kauran diterpene derivative and its application in antitumor drug is prepared Download PDFInfo
- Publication number
- CN108047037A CN108047037A CN201711474788.XA CN201711474788A CN108047037A CN 108047037 A CN108047037 A CN 108047037A CN 201711474788 A CN201711474788 A CN 201711474788A CN 108047037 A CN108047037 A CN 108047037A
- Authority
- CN
- China
- Prior art keywords
- formula
- compound
- organic layer
- reaction
- prepared
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/14—Preparation of carboxylic acid esters from carboxylic acid halides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/02—Esters of acyclic saturated monocarboxylic acids having the carboxyl group bound to an acyclic carbon atom or to hydrogen
- C07C69/12—Acetic acid esters
- C07C69/21—Acetic acid esters of hydroxy compounds with more than three hydroxy groups
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/07—Optical isomers
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
A kind of corresponding kauran diterpene derivative and its application in antitumor drug is prepared, shown in the structure such as formula (I) of the corresponding kauran diterpene derivative；The preparation method of formula (I) described compound, comprises the following steps, 1) by Isoforretin A, DMAP is dissolved in dichloromethane, triethylamine is added in, chloroacetic chloride is added dropwise at 0 DEG C~5 DEG C of heat preservation, reaction solution is warming up to 20 DEG C~25 DEG C after finishing and is stirred to react by drop；2) to TLC detections after the reaction was complete, into reaction solution adding in saturated sodium bicarbonate aqueous solution is quenched reaction, and with dichloromethane aqueous layer extracted three times, merging organic layer, organic layer is washed three times with dilute hydrochloric acid, three layers of saturated common salt water washing.Collected organic layer is dried with anhydrous magnesium sulfate, and filtering obtains crude product after concentration.Crude product purifies to obtain sterling target product formula (I) compound through flash chromatography on silica gel.
Description
Technical field：
The present invention relates to a kind of new Diterpenes chemical combination and its applications in antitumor drug is prepared.
Background technology：
Thioredoxin -1 ((thioredoxin-1, abbreviation Trx-1), it is abnormal in many tumour cells and tumor tissues
Height expression, the Trx1 of overexpression can promote tumor cell proliferation, by helping tumour cell with protein bindings such as ASK1, PTEN
Escape, Trx1 can be by increasing HIF alpha expressions, enhancing VEGF function promotion angiogenesis.In addition, clinical research shows Trx1 tables
Higher up to measuring, tumour patient prognosis is poorer.In addition to tumour, thing of many diseases all with the mediation of above-mentioned thioredoxin system
Part number | the reaction of the abnormal cell of hair is related..Therefore, the important need that new Trx1 selective depressants are medicine research and development are screened
It asks, they may be the drug of extremely effective treatment tumour and the disease of other Trx1 mediations.Chinese patent application
CN106631804A discloses a kind of compound for extracting from Rabdosia plant such as following formula
(Isoforresin A, abbreviation Iso A), the compound are a kind of correspondence-kauran diterpene derivative, are pointed out in this application
IsoA has certain inhibition to Partial tumors cell, and can extend the life cycle of liver cancer tumor-bearing mice, however from experiment
The result shows that the anticancer spectrum of IsoA is relatively narrow, only to liver cancer etc., a few oncotherapy is with obvious effects.Therefore on this basis into
Row, which improves a kind of correspondence-kauran diterpene derivative to different tumours with inhibition of offer, to be become in the prior art urgently
Problem to be solved.
The content of the invention
To improve the active anticancer of correspondence-kauran diterpene derivative, our passing through on the basis of existing Iso A
It learns modification and obtains a kind of new compound, and have surprisingly found that and can significantly improve its anticancer effect.
A kind of correspondence-kauran diterpene is to provide for solution aforementioned technical problem the technical solution adopted by the present invention to spread out
Biology, it is characterised in that shown in the structure such as formula (I) of the correspondence-kauran diterpene derivative
The present invention also provides the preparation methods of formula (I) compound, it is characterised in that comprise the following steps,
1) by Isoforretin A, DMAP is dissolved in dichloromethane, adds in triethylamine, acetyl is added dropwise at 0 DEG C~5 DEG C of heat preservation
Reaction solution is warming up to 20 DEG C~25 DEG C after finishing and is stirred to react by chlorine, drop.
2) to TLC detections after the reaction was complete, into reaction solution adding in saturated sodium bicarbonate aqueous solution is quenched reaction, uses dichloro
Methane aqueous layer extracted three times, merges organic layer, and organic layer is washed three times with dilute hydrochloric acid, three layers of saturated common salt water washing.Collection has
Machine layer is dried with anhydrous magnesium sulfate, and filtering obtains crude product after concentration.Crude product purifies to obtain sterling target product through flash chromatography on silica gel
Formula (I) compound；
Isoforretin A, DMAP, the molar ratio of triethylamine and chloroacetic chloride are 1:0.15~0.25：2~4：2~
4。
The present invention also provides application of the compound in antitumor drug is prepared as shown in formula (I).
The present invention also provides application of the compound in resistive connection rectal neoplasm drug is prepared as shown in formula (I).
The present invention also provides a kind of pharmaceutical composition, it is characterized in that described pharmaceutical composition includes chemical combination shown in formula (I)
Object.
The present invention also provides a kind of pharmaceutical preparation, formula (I) compound comprising therapeutically effective amount and can pharmaceutically connect
The excipient received.
Under study for action it was found that by Iso A carry out it is acylated after obtained formula (I) compound, can significantly improve pair
The external inhibitory activity of Trx1 in the body outer suppressioning experiment to tumour cell, shows colorectal cancer cell etc. more preferably
Tumor killing effect, it is based on body outer suppressioning experiment as a result, in the inhibiting tumor assay to colorectal cancer tumor-bearing mice, using formula (I)
The experiment mice of compound administration shows better tumor killing effect compared with using the experiment mice of similary dosage Iso A,
Illustrate that compound shown in formula (I) after chemical modification, is significantly improving the inhibition to colorectal cancer.
Specific embodiment
The synthesis of 1 formula of embodiment (I) compound
1) by Isoforretin A, DMAP (4-dimethylaminopyridine) is dissolved in dichloromethane, adds in triethylamine, keeps the temperature 0 DEG C
Corresponding chloroacetic chloride is added dropwise at~5 DEG C, reaction solution is warming up to 20 DEG C~25 DEG C after finishing and is stirred to react by drop.
2) to TLC detections after the reaction was complete, into reaction solution adding in saturated sodium bicarbonate aqueous solution is quenched reaction, uses dichloro
Methane aqueous layer extracted three times, merges organic layer, and organic layer is washed three times with dilute hydrochloric acid, three layers of saturated common salt water washing.Collection has
Machine layer is dried with anhydrous magnesium sulfate, and filtering obtains crude product after concentration.Crude product purifies to obtain sterling target product through flash chromatography on silica gel
Formula (I) compound；
Isoforretin A, DMAP, the molar ratio of triethylamine and chloroacetic chloride are 1:0.2：2.
To product carry out elemental analysis result be：C, 62.49；H, 6.99；O, 30.52.
The chemical formula of product is C30H40O11
13C-NMR(CDCl3) result such as following table
| The position of C | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
| 13C-NMR | 77.7 | 38.4 | 48.4 | 40.1 | 40.3 | 69.4 | 68.8 | 38.4 | 49.3 |
| The position of C | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 |
| 13C-NMR | 43.3 | 30.5 | 33.7 | 79.4 | 76.7 | 152.1 | 208.8 | 116.4 | 19.7 |
| The position of C | 19 | 20 | 21 | 22 | 23 | 24 | 25 | 26 | 27 |
| 13C-NMR | 23.6 | 23.6 | 170.2 | 21.0 | 170.2 | 21.0 | 170.2 | 21.0 | 170.2 |
| The position of C | 28 | 29 | 30 | ||||||
| 13C-NMR | 21.0 | 170.2 | 21.0 |
1 formula of Pharmacological Examples (I) compound react in vitro in Trx1 activity inhibitions
Experiment material:
Trx1 albumen is purchased from Sino Biological Inc., and Trx1 activity detection kits are purchased from the U.S.
Caymen Chemical companies.
Experimental method:
1) provide experimental method according to kit and prepare the detection liquid of Trx 1；
2) it is separately added into formula (I) compound (0-100 μM of final concentration) being serially diluted；
3) 37 DEG C of incubation 30min；
4) 5 μ L NADPH, 37 DEG C of incubation 10min are added in per hole；
5) 10 μ l fluorogenic substrates, 37 DEG C of incubation 5min are added in per hole；
6) microplate reader detects
7) Trx activity test methods under enzyme activity conditions in vitro are calculated:
Trx Activity determinations use Thioredoxin Activity Fluorescent Assay Kit under conditions in vitro
(Caymen Chemica l) is carried out.According to the requirement of kit specification, add in 2 μ L's in 0.02 μM of Trx samples
β-the NADPH of formula (I) compound (0.1 μM of 100 μM of to) of various concentration, then 5 μ L of each each addition of sample, 37 DEG C of incubations
30min. is eventually adding fluorogenic substrate, 30 to 60 minutes sections is being reacted, by microplate reader (Thermo Scientific
Varioskan Flash fluorescent microplate reader) record 518nm launch wavelength (488nm excitations)
To calculate Trx1 activity.
Experimental result see the table below
The experimental results showed that formula (I) compound starts to show that the apparent of Trx1 is inhibited to live when concentration reaches 0.5 μM
Property (enzyme activity 84.3%), and with concentration increase the rejection ability of Trx1 activity is gradually increased.It is right compared with IsoA
The inhibitory activity of Trx1 significantly improves.
2 formula of Pharmacological Examples (I) compound is to the inhibition of different types of solid tumor cell
Formula (I) compound (purity ＞ 99%) is dissolved in DMSO, configures 100uM mother liquors, -20 DEG C of preservations.Ten kinds of selection is often
See malignant entity tumor (including lung cancer, stomach cancer, breast cancer, colorectal cancer, liver cancer, cervical carcinoma, the cancer of the esophagus, carcinoma of mouth, prostate
Cancer, cancer of pancreas), the correspondence tissue normal cell of the tumor cell line of above-mentioned ten kinds of tumours of in vitro culture and wherein five kinds tumours
System, it is as follows：
Lung cancer：The normal pulmonary epithelial cells HBEC of people, human lung cancer cell A549, SK-MES-1
Stomach cancer：People normal gastric mucosa epithelial cell RGM-1, gastric carcinoma cells MGC-803, SGC-7901
Breast cancer：People normal mammary epithelial MCF10A, human breast cancer cell MCF7, MDA-MB-231
Colorectal cancer：People normal Colon and rectum mucomembranous cell FHC, Human colorectal cancer cells Caco-2, LOVO2
Liver cancer：Human normal hepatocyte LO2, human liver cancer cell HepG2, Hep3B
Cervical carcinoma：Human cervical carcinoma cell Hela
The cancer of the esophagus：Human esophagus cancer cell EC109
Carcinoma of mouth：Human tongue cancer cell line SCC9
Prostate cancer：Human Prostate Cancer Cells PC3
Cancer of pancreas：Human pancreatic cancer cell PANC-1
For any of the above cell line, handled for 24 hours using formula (I) compound of various concentration, then detected by mtt assay
Its inhibition to different cells calculates IC50, and the results are shown in table below
| Cell line | HBEC | RGM-1 | MCF10A | FHC | LO2 |
| IC50(μM) | 127.8 | 158.1 | 102.5 | 149.7 | 124.6 |
| Cell line | A549 | SK-MES-1 | MGC-803 | SGC-7901 | MCF7 |
| IC50(μM) | 32.8 | 45.1 | 36.7 | 40.9 | 15.9 |
| Cell line | MDA-MB-231 | Caco-2 | LOVO2 | HepG2 | Hep3B |
| IC50(μM) | 19.4 | 4.7 | 5.2 | 17.2 | 20.3 |
| Cell line | Hela | EC109 | SCC9 | PC3 | PANC-1 |
| IC50(μM) | 46.7 | 38.5 | 50.9 | 42.8 | 18.4 |
The results show：1) formula (I) compound is smaller to the toxicity of multigroup normal cell system, IC50 100-160uM；2. formula
(I) compound is relatively common to the cell line inhibition of lung cancer, stomach cancer, cervical carcinoma, the cancer of the esophagus, carcinoma of mouth and prostate cancer,
IC50 is 20-60uM, and 3. formula (I) compound has the cells such as colorectal cancer, breast cancer, liver cancer, cancer of pancreas preferable inhibit
Act on IC50 ＜ 20uM；4. formula (I) compound is very prominent to the inhibition of colorectal cancer cell (Caco-2, LOVO2),
IC50 is down to 4-5uM.Therefore, research object of the colorectal cancer as further zoopery is selected.
Pharmacological Examples 3, to the contrast test of the life cycle of tumor-bearing mice
1. material
A) cell line:The colorectal cancer cell Caco-2 of expressing luciferase can be stablized, trained purchased from Chinese Academy of Sciences typical case
Yang Wu preservations committee cell bank.
B) animal:BALB/c nude mouses 30, weight (13 ± 2g), male.
C) drug:Iso A (drug 1), formula (I) compound (drug 2)
2. experimental method
A) making of colorectal cancer bearing mouse model:Take the logarithm growth period colorectal cancer cell Caco-2, pancreatin digestion
After be configured to 1x107Cells/mL cell suspensions, lmL syringes inoculation 200 μ L (+100 μ l matrigels of 100 μ l cells) cell
Suspension is in the right side oxter of nude mice (normal group only injects 200 μ L matrigels).
B) experimental animal grouping and administration:Raising observes lotus knurl in Mice Body by IVIS small animal imagings system after two weeks
Situation takes and builds successfully (gross tumor volume >=100mm3) tumor-bearing mice it is spare, every group 10 (every group of average external volume is consistent, swell
Knurl volume is excessive or too small person rejects).Daily intraperitoneal injection once, continuous three weeks.A tumor size is recorded every three days,
So that it is determined that therapeutic effect.Experiment terminates to put to death experimental animal, detects tumor weight and calculates tumour inhibiting rate
Tumour inhibiting rate=[1- (experimental group be averaged knurl weight/control group be averaged knurl weight)] × 100%.
Specific grouping and administration such as following table
Detect the different dosing number of days of each group experimental animal gross tumor volume, it is dead after tumor weight and life cycle
Data add following table, and carry out t inspections to result, and experimental result see the table below (means ± s, n=10)
| Group | Model | Experiment 1 | Experiment 2 |
| 0d/mm3 | 109±7.27 | 104±7.25 | 88±6.16 |
| 3d/mm3 | 124±8.27 | 117±8.55 | 93±9.01 |
| 6d/mm3 | 435±42.05 | 400±38.69 | 300±29.01 |
| 9d/mm3 | 693±46.20 | 624±45.74 | 437±36.38 |
| 12d/mm3 | 835±58.45 | 735±71.03 | 463±30.86 |
| 15d/mm3 | 1170±113.10 | 1006±100.62 | 584±52.52 |
| 18d/mm3 | 1480±118.40 | 1258±83.87 | 642±44.91 |
| 21d/mm3 | 1805±138.38 | 1534±107.40 | 690±57.53 |
| Average knurl weight/g | 1.53 | 1.30 | 0.58 |
| Tumour inhibiting rate % | 15% | 62% |
The result shows that formula (I) compound can have the tumour growth for significantly inhibiting colorectal cancer tumor-bearing mice, and same
It waits under dosage, the opposite experimental group 1 for using IsoA, experimental group 2 also can the more significant life cycle (P for significantly extending tumor-bearing mice
＜ 0.01).Example of formulations
Formula (I) compound is made by embodiment 1, then tablet is made as follows:Every composed of the following components
Formula (I) compound 100mg, appropriate starch, appropriate corn pulp, Magnesium Stearate proper quantity, by above-mentioned each component according to normal
Advising tablet production method and making becomes tablet.
Claims (6)
- A kind of 1. correspondence-kauran diterpene derivative, it is characterised in that the structure of the correspondence-kauran diterpene derivative As shown in formula (I)
- 2. the preparation method of compound as described in formula (I) in claim 1, it is characterised in that comprise the following steps,1) by Isoforretin A, DMAP is dissolved in dichloromethane, adds in triethylamine, chloroacetic chloride is added dropwise at 0 DEG C~5 DEG C of heat preservation, drip Reaction solution is warming up to 20 DEG C~25 DEG C after finishing and is stirred to react；2) to TLC detections after the reaction was complete, into reaction solution adding in saturated sodium bicarbonate aqueous solution is quenched reaction, uses dichloromethane Aqueous layer extracted three times, merges organic layer, and organic layer is washed three times with dilute hydrochloric acid, three layers of saturated common salt water washing.Collected organic layer It is dried, filtered with anhydrous magnesium sulfate, crude product is obtained after concentration.Crude product purifies to obtain sterling target product formula through flash chromatography on silica gel (I) compound；Isoforretin A, DMAP, the molar ratio of triethylamine and chloroacetic chloride are 1:0.15~0.25：2~4：2~4.
- 3. application of the compound in antitumor drug is prepared as shown in formula (I) in claim 1.
- 4. application of the compound in resistive connection rectal neoplasm drug is prepared as shown in formula (I) in claim 1.
- 5. a kind of pharmaceutical composition, it is characterized in that described pharmaceutical composition includes compound shown in formula (I).
- 6. a kind of pharmaceutical preparation, it is characterized in that formula (I) compound and pharmaceutically acceptable figuration comprising therapeutically effective amount Agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711474788.XA CN108047037A (en) | 2017-12-29 | 2017-12-29 | A kind of correspondence-kauran diterpene derivative and its application in antitumor drug is prepared |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711474788.XA CN108047037A (en) | 2017-12-29 | 2017-12-29 | A kind of correspondence-kauran diterpene derivative and its application in antitumor drug is prepared |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108047037A true CN108047037A (en) | 2018-05-18 |
Family
ID=62128959
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711474788.XA Pending CN108047037A (en) | 2017-12-29 | 2017-12-29 | A kind of correspondence-kauran diterpene derivative and its application in antitumor drug is prepared |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108047037A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108129319A (en) * | 2017-12-29 | 2018-06-08 | 天津市人民医院 | A kind of Isoforrensin A derivatives and its application in preparation of anti-tumor drugs |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106631804A (en) * | 2016-12-21 | 2017-05-10 | 江苏省中医药研究院 | Compound separated out from labiatae isodon plant, and preparation method and application thereof |
| CN108129319A (en) * | 2017-12-29 | 2018-06-08 | 天津市人民医院 | A kind of Isoforrensin A derivatives and its application in preparation of anti-tumor drugs |
-
2017
- 2017-12-29 CN CN201711474788.XA patent/CN108047037A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106631804A (en) * | 2016-12-21 | 2017-05-10 | 江苏省中医药研究院 | Compound separated out from labiatae isodon plant, and preparation method and application thereof |
| CN108129319A (en) * | 2017-12-29 | 2018-06-08 | 天津市人民医院 | A kind of Isoforrensin A derivatives and its application in preparation of anti-tumor drugs |
Non-Patent Citations (3)
| Title |
|---|
| YU ZHAO 等: "Synthesis and cytotoxicity of some new eriocalyxin B derivatives", 《EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY》 * |
| 王琳: "《有机化学反应（三）》", 31 December 1997, 化学工业出版社 * |
| 闫学斌: "贝壳杉烷型二萜类化合物的结构、衍生物设计合成及生物活性的研究", 《中国优秀博硕士学位论文全文数据库 (博士) 工程科技Ⅰ辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108129319A (en) * | 2017-12-29 | 2018-06-08 | 天津市人民医院 | A kind of Isoforrensin A derivatives and its application in preparation of anti-tumor drugs |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107721959A (en) | Myricetin derivative and preparation method, and its application in treatment colitis, the conversion of preventing and treating colitis cancer and treatment colorectal cancer | |
| CN110290794A (en) | The microRNA and its purposes in cancer treatment of 5- halo uracil modification | |
| CN104387389A (en) | 1,2,3-Triazole-flavonoid compound-sophocarpidine ternary conjugate and use | |
| CN106749089A (en) | The preparation of new fluoro thiazole hydrazone compounds and its application in antineoplastic | |
| CN106963752A (en) | A kind of antineoplastic containing Histone deacetylase inhibitor derivates | |
| CN104367584A (en) | Application of doxycycline in preparation of antitumor drugs | |
| CN108047037A (en) | A kind of correspondence-kauran diterpene derivative and its application in antitumor drug is prepared | |
| CN104557909B (en) | 3- acyloxy replaces dextrorotation deoxidation tylophorinine derivative, its preparation method and pharmaceutical composition and purposes | |
| CN108129319A (en) | A kind of Isoforrensin A derivatives and its application in preparation of anti-tumor drugs | |
| CN106974908A (en) | Pharmaceutical composition and purposes containing hdac inhibitor and IRE1 inhibitor | |
| CN102234258A (en) | Preparation method and application of sphaelactone | |
| CN106748939A (en) | The new bromine phenol thiosemicarbazide compound of one class and its preparation and medicine and purposes | |
| CN105213366B (en) | The medical usage and its pharmaceutical composition of gamboge ketone compound | |
| CN109876154A (en) | The nanoparticle preparation of a kind of polysaccharides modification and its antitumor activity | |
| CN105017352B (en) | A kind of indole alkaloid glucoside and its application | |
| CN110314149B (en) | Poly-pregnen-zinc tablet and preparation process thereof | |
| CN103316035A (en) | Use of sodium bicarbonate in preparation of medicines for treating cancer | |
| CN104447921A (en) | Arctigenin-gemcitabine conjugate as well as preparation method and medical application thereof | |
| CN1839865A (en) | Antineoplastic baicalein and baicalin synergistic pharmaceutical composition | |
| CN103351383B (en) | 5-fluorouracil nitroxyl-free-radical anti-tumor drug | |
| CN103130871B (en) | Preparation method and application of prodrug of endopeptidase activated doxorubicin | |
| CN108047043A (en) | It is isolated from compound and its application of Rabdosia plant | |
| CN105541696B (en) | A kind of antitumor compound and its preparation method and application | |
| CN104983921A (en) | Anti-cancer and anti-mutation combination component and preparation method thereof | |
| CN104725291B (en) | XPO1 protein inhibitors |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | 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: 20180518 |