CN105439859A - New monoterpene compound and preparation method and application thereof - Google Patents
New monoterpene compound and preparation method and application thereof Download PDFInfo
- Publication number
- CN105439859A CN105439859A CN201510875223.7A CN201510875223A CN105439859A CN 105439859 A CN105439859 A CN 105439859A CN 201510875223 A CN201510875223 A CN 201510875223A CN 105439859 A CN105439859 A CN 105439859A
- Authority
- CN
- China
- Prior art keywords
- compound
- component
- formula
- preparation
- column chromatography
- 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.)
- Granted
Links
Classifications
-
- 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
- C07C69/16—Acetic acid esters of dihydroxylic compounds
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
- C07C67/56—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a monoterpene compound (please see the structural formula in the specification). The molecular formula of the compound is C12H18O4. The unsaturation degree of the compound is 4. The ultraviolet spectrum shows that the maximum ultraviolet absorption of the compound is 203 nm. The invention further relates to an extraction and separation method of the compound. Extraction is conducted through ethyl acetate, extraction liquid is separated and purified, and then the monoterpene compound is obtained. Due to the activity effect of the compound on resisting fungi, the lead compound is provided for development of medicine for resisting fungi, and important significance in development of new application of natural products and plant medicine is achieved.
Description
Technical field
The invention belongs to organic compound technical field, be specifically related to a kind of new monoterpenes compound and its preparation method and application.
Background technology
Tibet Paeonia ludlowii (Paeonialudlowii) is Ranunculaceae (Ranunculacea), Paeonia (Paeonia), Section Moutan (Sect.MoutanDC.) plant.In the open river valley, the Yarlung Zangbo River only growing the height above sea level 2500 ~ 3500m in the county such as Milin, Linzhi, Bomi, Chayu, Longzi in China Tibet and hillside border shrubbery; quantity is few; and in reducing trend year by year, belong to the distinctive endangered protective plant in Tibet.Record according to local local chronicle, Tibetan just used Tibet Paeonia ludlowii root skin treatment gynaecopathia, cardiovascular and cerebrovascular diseases, tinea etc. before the centuries.
At present, correlative study is had to the medicinal effect of Paeonia ludlowii root skin, but because in Paeonia ludlowii root skin, chemical composition is varied, research for its activeconstituents yet there are no relevant report, applicant utilizes normal phase silica gel column chromatography, anti-phase (MCI-Gel) column chromatography, gel (SephadexLH-20) column chromatography, the ethyl acetate extraction components of the technology such as high performance liquid chromatography to Paeonia ludlowii root skin is separated, purifying, obtain monomeric compound 23, pass through UV spectrum, infrared spectra, mass spectrum, nuclear magnetic resonance spectrum also determines its structure in conjunction with bibliographical information, comprising monoterpenes compound 7, monoterpene glycosides compound 3, phenolic compound 8, phenolic glycoside compounds 3, alkaloid compound 1, triterpene compound 1, wherein new compound 4, known compound 19, be separated from the Paeonia ludlowii of Tibet all first and obtain.
Summary of the invention
The present invention, by the further investigation to Paeonia ludlowii root skin, provides a kind of new compound and its preparation method of monoterpenes.
The present invention realizes especially by following technical scheme:
A kind of monoterpenes compound, its structural formula is such as formula shown in I:
Compound described in formula I is colorless oil, by ESI-MS spectrum ([M+Na]
+m/z249) and HR-ESI-MS spectrum ([M+Na]
+m/z249.1101, calcdforC
12h
18o
4na, 249.1097), determine the molecular formula C of this compound
12h
18o
4, its degree of unsaturation is 4.Infrared spectra (KBr compressing tablet) gives hydroxyl (3435cm
-1) and carbonyl (1720cm
-1) signal.UV spectrum shows this compound ultraviolet maximum absorption at 203nm.
The preparation method of described formula I, specifically comprises the following steps:
1), after getting dry Tibet Paeonia ludlowii root skin pulverizing, add 95% ethanol, room temperature lixiviate 24h, repeat suction filtration, 50 DEG C of concentrating under reduced pressure, utilize ethyl acetate to enter extraction, by extraction liquid concentrating under reduced pressure under 50 DEG C of conditions, obtain dry powder A;
2) ratio of dry powder A and polymeric amide 1:4 ~ 5 is in mass ratio mixed, utilize volumetric concentration be 40% methanol aqueous solution wash-out obtain B component;
3) B component is used 120mL dissolve with methanol, silica gel mixed sample, treat that solvent volatilizees completely, carry out silica gel column chromatography, be the mixed solution wash-out of 10 ︰ 1 by sherwood oil/Virahol volume ratio, obtain component C;
4) component C is repeated silica gel column chromatography, be the mixed solution wash-out of 9 ︰ 1 by sherwood oil and Virahol volume ratio, obtain component D;
5) component D is continued through gel column chromatography, methyl alcohol, as elutriant, obtains formula I monomeric compound.
In preparation method's step (1) of the present invention, the mass volume ratio of root skin and 95% ethanol is 1:10.
In preparation method's step (2) of the present invention, the ratio of dry powder A and the preferred 1:4.2 in mass ratio of polymeric amide mixes.
Present invention also offers the application of formula I as antifungal drug.
Described medicine can be processed into any pharmaceutically acceptable formulation on demand, and wherein more preferably formulation is tablet, capsule, granule, syrup, lyophilized injectable powder or injection liquid.Its preparation can be prepared by common working method known in those skilled in the art, after mixing with liquid solvent or solid carrier by formula I, then add one or more in weighting agent, tackiness agent, wetting agent, disintegrating agent, absorption enhancer, solvent, surfactant, flavouring agent, sanitas, lubricant, sweeting agent or pigment.
Medicine of the present invention can by user before use through diluting or directly using.
Beneficial effect of the present invention is: the present invention's extraction and isolation from the Paeonia ludlowii root skin of Tibet goes out a kind of new compound, and this Extraction and isolation method is easy, quick, and efficiency is high.This compound has antimycotic active function simultaneously, compares Tibet Paeonia ludlowii root skin pharmacological action more definite, solves complex chemical composition in the Paeonia ludlowii root skin of Tibet, the problem that difficult quality controls.
Embodiment
Below in conjunction with embodiment, the present invention is described further, the following stated, only to preferred embodiment of the present invention, not do other forms of restriction to the present invention, any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed to the Equivalent embodiments of equal change.Everyly do not depart from the present invention program's content, any simple modification done following examples according to technical spirit of the present invention or equivalent variations, all drop in protection scope of the present invention.
Embodiment 1
Get dry Tibet Paeonia ludlowii root skin, accurately 10kg is taken after pulverizing, 95% ethanol is added according to the ratio of Gen Pi and solvent 1:10 (W/V), room temperature lixiviate 24h, suction filtration, in triplicate, 50 DEG C of concentrating under reduced pressure, be extracted with ethyl acetate by total extract subsequently, extraction liquid is concentrating under reduced pressure under 50 DEG C of conditions, obtains the component dry powder A of 356g.
Dry powder A is taken 356g and 1.5kg polymeric amide and mixes sample, carry out medium pressure liquid chromatography, reversed-phase column packing material is MCI-Gel, utilize volumetric concentration be 40% methanol aqueous solution wash-out wash-out obtain 57g B component, use 120mL dissolve with methanol, silica gel mixed sample, treat that solvent volatilizees completely, carry out silica gel column chromatography, with the mixed solution wash-out that sherwood oil/Virahol volume ratio is 10 ︰ 1, obtain component C, component C is repeated silica gel column chromatography, with the mixed solution wash-out that sherwood oil and Virahol volume ratio are 9 ︰ 1, obtain component D, component D is continued through gel column chromatography, methyl alcohol is as elutriant, obtain formula I monomeric compound 7mg.
Embodiment 2 monomeric compound Structural Identification
1) wave spectrum test
Be separated the compound obtained, liquid chromatography carries out purity detecting by analysis, if without uv-absorbing, analyzing liquid chromatography cannot detect, and carries out TLC detection.And choose the dissolving of corresponding deuterated solvent according to monomeric compound polarity, utilize NMR to detect, tetramethylsilane (TMS) is as interior mark.New compound needs KBr compressing tablet to carry out functional group's kind contained in IR authenticating compound structure; Meanwhile, get appropriate compound, dissolve with corresponding polar solvent and carry out UV detection, there is in deterministic laminate structures functional group's kind of uv-absorbing.Compound molecular weight is inferred according to ESI-MS, more further according to the molecular weight of HR-ESI-MS deterministic compound, molecular formula and degree of unsaturation.
2) determination of sugared configuration
With reference to acid hydrolysis thin layer chromatography, take appropriate compound, dissolve with methanol, be mixed with the solution of 1mg/mL, with kapillary point sample on silica-gel plate, and be invested in and be equipped with in the airtight separation chamber of the vitriol oil, 60 DEG C of water-bath 30min, after terminating, taking-up blower blows to solvent and waves to the greatest extent, D-Glucose on silica-gel plate blank space point, D-celery sugar, D-xylose standard product, by chloroform/methanol (7:3, v/v) mixing solutions water saturation, get the chloroform/methanol solution after 9mL water saturation, the Glacial acetic acid adding 1mL again mixes and to launch as developping agent afterwards, spray with the butanol solution of phthalic acid aniline, in 105 DEG C of heating 10min colour developing.
The configuration of aniline-phthalic acid developer: accurately take aniline 0.93g and phthalic acid 1.66g and be dissolved in the water saturated propyl carbinol of 100mL.
3) results and analysis
Colorless oil.By ESI-MS spectrum ([M+Na]
+m/z249) and HR-ESI-MS spectrum ([M+Na]
+m/z249.1101, calcdforC
12h
18o
4na, 249.1097), determine the molecular formula C of this compound
12h
18o
4, its degree of unsaturation is 4.IR spectrum (KBr compressing tablet) gives hydroxyl (3435cm
-1) and carbonyl (1720cm
-1) signal.UV spectrum shows this compound ultraviolet maximum absorption at 203nm.
In 1H-NMR spectrum, give 3 methyl hydrogen signal δ H1.23 (s, 3H, H-8), δ H1.15 (d, J=6.6Hz, 3H, H-10) and 2.05 (s, 3H, H-12); 3 methylene radical hydrogen signal δ H1.87 (dd, J=9.0,18.6Hz, 1H, H-3a), δ H2.24 (dd, J=9.1,18.6Hz, 1H, H-3b), δ H1.16 (dd, J=3.6,4.8, Hz, 1H, H-7a), δ H1.49 (dd, J=4.8,9.6Hz, 1H, H-7b) and 4.05 (d, J=11.4Hz, 1H, H-9a), 4.29 (d, J=11.4Hz, 1H, H-9b); 2 methyne hydrogen signal δ H2.51-2.84 (m, 1H, H-2) and δ H1.94 (dd, J=3.0,9.6Hz, 1H, H-5).In 13C-NMR spectrum, give 12 carbon signals altogether, obtain in conjunction with DEPT spectrum and 1H-NMR spectrum: 1 ketone carbonyl, 1 ester carbonyl group, the quaternary carbon of 2 sp3 hydridization, the methyne of 2 sp3 hydridization, the methylene radical of 3 sp3 hydridization, and 3 methyl.The prompting of 1H-1HCOSY spectrogram has the existence of 2 fragments: a:-CH2CH-; B:-CH2CHCH3-.
Analyze HMBC compose and compose in conjunction with its hydrogen, carbon modal data and compound
relatively, have identical carbon skeleton, difference is that on C-9, hydroxyl is acetylation.Composed by HMBC, δ H4.05 (d, J=11.4Hz; 1H, H-9a), δ H4.29 (d, J=11.4Hz; 1H; H-9b) with δ C172.71 (s, C-11), δ H2.05 (s; 3H; H-12) be obviously correlated with δ C172.71 (s, C-11), show that on C-9, hydroxyl is acetylation.Therefore,
Finally determine that this compound structure is
nuclear magnetic data is in table 1.
The hydrogen spectrum of table 3-2 compound F 17-hydroxy-corticosterone rp34 and carbon modal data (MeOD)
Embodiment 3 anti-mycotic activity measures
Microsporon gypseum is the filamentous fungus that a class can cause shallow layer skin dermatophytosis, and can cause ringworm of the body, jock itch, onychomycosis, favus of the scalp etc., be clinical common superficial part pathomycete.The effect of medicinal plant in treatment tetter in the last few years received much concern, seek and utilize the activeconstituents in natural medicinal plant to design Novel non-toxic to treat dermopathic medicine be an important research direction.Minimal inhibitory concentration (Minimuminhibitoryconcentration, MIC) weighs the important indicator of the active size of antibacterials, adopts test tube medicine base method, not only can interpretation MIC value easily, and easy economy, easily operates.
" filamentous fungi trace broth dilution method drug sensitivity test scheme " (M38-A2) research that this laboratory reference Clinical Laboratory Standard association (CLSI) is recommended is separated the activity of the monomeric compound obtained.
1. test monomeric compound
Embodiment 1 is separated the formula I obtained.
2. test strain
Microsporon gypseum bacterial strain is purchased from Chinese Academy of Medical Sciences's Nanjing Hospital of Skin Diseases.
3. test apparatus
4. test reagent
5. the preparation of substratum
Aseptically, get fresh modified form 1640 (containing L-glutaminate not carbonate containing) substratum 1000mL, with 3-(N-morpholinyl)-propanesulfonic acid damping fluid that concentration is 0.165mol/L, Medium's PH Value is adjusted to 7.0, then use the filtering with microporous membrane of 0.22 μm degerming and packing, 4 DEG C save backup.
6. the preparation of bacteria suspension
Aseptically, get eugonic microsporon gypseum test tube slant and add stroke-physiological saline solution, with the thread culture of transfering loop scraping media surface gently, render in the mortar doing aseptically process and grind, with blood cell counting plate counting, concentration improvement 1640 liquid nutrient medium of bacteria suspension is adjusted to 0.5 × 10
3-2 × 10
3cFU/mL is for subsequent use.
7. test method
The preparation of drug sensitive plate
Accurately take monomeric compound and the griseofulvin tablet powder of certain mass respectively, add the DMSO of certain volume and content is less than 1%, be mixed with the storing solution of 0.8mg/mL, then storing solution be diluted to 2 times of final concentrations by 1:25, i.e. 32 μ g/mL with 1640 liquid nutrient mediums after improvement.The 1st hole of the every a line of 96 orifice plate is added with the liquid that pipettor gets the 32g/mL that 200 μ L have prepared respectively; 2-11 hole adds 1640 liquid nutrient mediums after 100 μ L improvement respectively; Get 100 μ L liquids with pipettor from the 1st hole and inject the 2nd hole, 2 times are diluted to the 10th hole successively, and the 100 μ L liquid rejects will got from the 10th hole, finally obtain 1-10 hole liquor strength and be respectively 32 μ g/mL, 16 μ g/mL, 8 μ g/mL, 4 μ g/mL, 2 μ g/mL, 1 μ g/mL, 0.5 μ g/mL, 0.25 μ g/mL, 0.125 μ g/mL, 0.0625 μ g/mL, ten gradient concentrations; Then 1-11 hole adds 100 μ L bacteria suspensions respectively, and the 11st hole is as growth control, and the 12nd hole adds improvement 1640 liquid nutrient medium of 200 μ L as blank; Three repetitions established by each sample, and each sample establishes growth control and blank.
Antibacterial result judges
The drug sensitive plate inoculating bacteria suspension is placed in 28 (± 1) DEG C constant incubator to cultivate, takes out after 7 days, the MIC value of monomeric compound to this bacterial classification is the drug level of 100% Developing restraint, and namely visual inspection is clarified or had no growth.
8. result
In order to find monomeric compound microsporon gypseum in the Paeonia ludlowii root skin ethyl acetate layer of Tibet to resistance, we utilize the new monomeric compound obtained from ethyl acetate layer Component seperation to carry out microsporon gypseum resistant proof, test-results measures and learns that embodiment 1 is separated the MIC value of formula I to microsporon gypseum obtained is 4 μ g/mL, compared with reference substance grisovin MIC0.5 μ g/mL, illustrate that this compound has certain restraining effect to microsporon gypseum.
Claims (6)
1. a new monoterpenes compound, is characterized in that: described structural formula of compound is such as formula shown in I:
2. monoterpenes compound according to claim 1, is characterized in that: described compound is colorless oil, and molecular formula is C
12h
18o
4, degree of unsaturation is 4.
3. the preparation method of compound according to claim 1, is characterized in that comprising the following steps:
1), after getting dry Tibet Paeonia ludlowii root skin pulverizing, add 95% ethanol, room temperature lixiviate 24h, repeat suction filtration, 50 DEG C of concentrating under reduced pressure, utilize ethyl acetate to enter extraction, by extraction liquid concentrating under reduced pressure under 50 DEG C of conditions, obtain dry powder A;
2) ratio of dry powder A and polymeric amide 1:4 ~ 5 is in mass ratio mixed, utilize volumetric concentration be 40% methanol aqueous solution wash-out obtain B component;
3) B component is used 120mL dissolve with methanol, silica gel mixed sample, treat that solvent volatilizees completely, carry out silica gel column chromatography, be the mixed solution wash-out of 10 ︰ 1 by sherwood oil/Virahol volume ratio, obtain component C;
4) component C is repeated silica gel column chromatography, be the mixed solution wash-out of 9 ︰ 1 by sherwood oil and Virahol volume ratio, obtain component D;
5) component D is continued through gel column chromatography, methyl alcohol, as elutriant, obtains formula I monomeric compound.
4. preparation method according to claim 3, is characterized in that: in step (1), the mass volume ratio of root skin and 95% ethanol is 1:10.
5. preparation method according to claim 3, is characterized in that: the ratio of dry powder A and polymeric amide 1:4.2 in mass ratio mixes in step (2).
6. formula I described in claim 1 is as the application of antifungal drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510875223.7A CN105439859B (en) | 2015-07-21 | 2015-12-02 | A kind of new monoterpenes compound and its preparation method and application |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510428171 | 2015-07-21 | ||
| CN2015104281719 | 2015-07-21 | ||
| CN201510875223.7A CN105439859B (en) | 2015-07-21 | 2015-12-02 | A kind of new monoterpenes compound and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105439859A true CN105439859A (en) | 2016-03-30 |
| CN105439859B CN105439859B (en) | 2018-05-18 |
Family
ID=55550563
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510875223.7A Expired - Fee Related CN105439859B (en) | 2015-07-21 | 2015-12-02 | A kind of new monoterpenes compound and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105439859B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948376A (en) * | 2010-09-19 | 2011-01-19 | 兰州佛慈制药股份有限公司 | Rapid extraction and separation method of paeonolum |
| CN103274919A (en) * | 2013-05-14 | 2013-09-04 | 亳州千草药业有限公司 | Extraction method of peony overground part |
| CN103524322A (en) * | 2013-10-24 | 2014-01-22 | 洛阳祥和牡丹科技有限公司 | Method for extracting paeonol from paeonia suffruticosa roots |
-
2015
- 2015-12-02 CN CN201510875223.7A patent/CN105439859B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948376A (en) * | 2010-09-19 | 2011-01-19 | 兰州佛慈制药股份有限公司 | Rapid extraction and separation method of paeonolum |
| CN103274919A (en) * | 2013-05-14 | 2013-09-04 | 亳州千草药业有限公司 | Extraction method of peony overground part |
| CN103524322A (en) * | 2013-10-24 | 2014-01-22 | 洛阳祥和牡丹科技有限公司 | Method for extracting paeonol from paeonia suffruticosa roots |
Non-Patent Citations (1)
| Title |
|---|
| XIA LI,ET AL.: "Two New Monoterpenes from Tithonia diversifolia and Their Anti-Hyperglycemic Activity", 《REC. NAT. PROD.》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105439859B (en) | 2018-05-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111139188B (en) | Novel skeleton heteroterpene derivative derived from marine fungi and application of novel skeleton heteroterpene derivative in preparation of anti-inflammatory drugs | |
| CN106946766B (en) | Alkaloid compound and its extraction separation method in purslane | |
| CN108277164B (en) | A kind of indene derivatives derived from endophytic fungi in Excoecaria agallocha and its application in preparing anti-inflammatory drugs | |
| CN101735236B (en) | Dimer citrinin compounds and preparation and use thereof | |
| CN103880808A (en) | Xanthone compounds and applications thereof | |
| CN102311981A (en) | Method for preparing and purifying prodigiosin | |
| CN108315264A (en) | A kind of polyketide in sea paint endogenetic fungus source and its application in preparing anti-inflammatory drug | |
| CN108017685A (en) | Novel triterpene saponin class compound and its preparation method and application | |
| CN106967024A (en) | A kind of α pyrone derivatives and its preparation method and application | |
| CN107973769B (en) | A kind of benzodihydropyrone class compound and its preparation method and application | |
| CN109134574A (en) | Steroidal compounds and the preparation method and application thereof and anti-tumor drug | |
| CN103880826B (en) | A kind of isobenzofuran ketonic compound and its preparation method and application | |
| CN104292237B (en) | A kind of six ring alkaloid compounds and preparation method and application | |
| CN105439859A (en) | New monoterpene compound and preparation method and application thereof | |
| CN114085272B (en) | Isaridin cyclic depsipeptide derivative and preparation method and application thereof | |
| CN105503602A (en) | Compound extracted from paeonia ludlowii and preparation method and application of compound | |
| CN109942658A (en) | A kind of miscellaneous terpene compound and the preparation method and application thereof and anti-tumor drug | |
| CN105481670A (en) | Pinane monoterpene compound, and preparation method and application thereof | |
| CN106478399B (en) | Derivative in hydroxy anthraquinones category and its application | |
| CN104804020A (en) | Sulfodionepiperazine compound, and preparation method and use thereof | |
| CN105439843A (en) | New compound and preparation method and application thereof | |
| CN110368389A (en) | The preparation method and applications and Claritin of phenolic compound | |
| CN1288163C (en) | Preparation process for compound of proto steroid soap oside and application thereof | |
| CN102659734B (en) | Triene antibiotic, preparation method thereof and application thereof | |
| CN114409625B (en) | Keratinone with neuroprotective activity and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180518 Termination date: 20201202 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |