CN102690262B - Lead compound of targeting human La protein and purpose thereof in preparation of medicament for resisting hepatitis B virus - Google Patents
Lead compound of targeting human La protein and purpose thereof in preparation of medicament for resisting hepatitis B virus Download PDFInfo
- Publication number
- CN102690262B CN102690262B CN201110071212.5A CN201110071212A CN102690262B CN 102690262 B CN102690262 B CN 102690262B CN 201110071212 A CN201110071212 A CN 201110071212A CN 102690262 B CN102690262 B CN 102690262B
- Authority
- CN
- China
- Prior art keywords
- protein
- lead compound
- virus
- preparation
- medicament
- 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention belongs to the field of pharmacy and relates to a lead compound of a targeting human La protein and purpose thereof in preparation of a medicament for resisting hepatitis B virus. According to the invention, research on enhancement of HBVRNA stability by the targeting La protein is employed to screen out the lead compound of the targeting human La protein shown as a formula (I), a formula (II) or a formula (III). Tests show that the lead compound has strong antiviral activity and effect of targeting the La protein. The lead compound of the present invention can be further optimized in structure and has good application prospect; and the lead compound and derivatives thereof can be used for preparation of medicament for resisting hepatitis B virus, as well as preparation of medicament targeting the La protein.
Description
Technical field
The invention belongs to pharmacy field, relate to the new pharmaceutical usage of the lead compound of target human La protein, the purposes of the lead compound that is specifically related to target human La protein in the medicine of preparing anti-hepatitis B virus.
Background technology
Known hepatitis B is one of transmissible disease of serious threat human health.According to U.S. Hollinger statistical report, hepatitis B virus is global distribution, approximately has in the world at present the chronic carrier of 3.5 hundred million HBV, approximately has 1,000,000 people to die from the disease that HBV causes every year.China belongs to the high epidemic regions that HBV infects.2007 " China Health statistical yearbook " showed, the sickness rate of China's viral hepatitis has been risen to 102.09/10 ten thousand people of 2006 by 64.91/10 ten thousand people of 2000, and Chronic Hepatitis B reaches more than 2,000 ten thousand people, accounts for 1.6% left and right of population; Wherein, hepatitis B virus surface antigen carrying rate is up to 9.8%, and total number of persons is up to people more than 100,000,000.In " Chinese the guideline ", adefovir ester, Entecavir and lamivudine are recommended as to a line medication of national hepatitis B virus resisting, these nucleosides and nucleotide analog, by suppressing copying of HBV DNA, reach the effect of alleviating or treating hepatitis B.
Along with widening and medication crowd's increase of medicinal application face, in succession report after the nucleoside medicine long-term prescriptions such as lamivudine both at home and abroad, exist patient compliance problem and many untoward reactions, its resistant rate has reached 69%; Due to the decline of described drug susceptibility, in minority immunodeficiency patient, there is the virus strain that HBV DNA polymerase gene is undergone mutation, cause body inner virus variation and sb.'s illness took a turn for the worse phenomenon, thereby limited its prolonged application.
In developing new drug, commonly use the method for virtual screening (virtual screening), the method filters out the chemical molecular that may have treatment effectiveness by computing method.At present widely used is that target (target) to medicine utilizes the experiment of X-ray diffraction or nucleus magnetic resonance (NMR), or uses the method for bioinformation structure prediction, obtains after the three-dimension high-resolution structure of target, then carries out virtual screening.The successful prerequisite of medicine virtual screening has three: (1) enough large chemical spaces (chemical space), (2) score function (scoring function) accurately, (3) high efficiency search algorithm method (searching algorithm), described chemical space, i.e. chemline used, broad sense, chemical space comprises the configuration (conformation) that part (ligand) and acceptor may exist, described score function, its design is mainly from protein structure database (Protein Databank, http://www.pdb.org) the middle combination (complex) of finding the good part of resolving power and acceptor, use the calculating of some energy, add the statistical method such as multiple linear regression (multivariate linear regression), set up a mathematical function accurately, and can fast and effeciently infer that whether stably unknown part to a given acceptor combination, even predict the free energy (binding free energy) of bonding strength (binding affinity) or combination.Above-mentioned score function is energy variation situation very complicated while doing translation (translation), rotation (rotation) or topographical variations (conformational changes) according to ligand molecular with respect to acceptor, finds the binding pattern with best score value.
Have research to point out, phosphorylation is the activity form of La albumen, and La albumen contributes to increase the stability of HBV RNA.Up to now, there is not yet about the biology for La protein binding Mechanism Design or the report of chemotherapeutic agent.
Summary of the invention
The object of this invention is to provide the new pharmaceutical usage of the lead compound of target human La protein, the purposes of the lead compound that is specifically related to target human La protein in the medicine of preparing anti-hepatitis B virus.
The present invention is increasing target La albumen on the Research foundation of HBV rna stability, screening obtains the lead compound of the target human La protein with new texture entity, and the lead compound of described target human La protein is the compound for the small molecule structure of La and RNA binding site.
In the present invention, described lead compound is HBSC11 or 15 or 34, has the structure of formula I or formula II or formula III,
HBSC11 HBSC15 HBSC34
（Ⅰ） （Ⅱ） （ Ⅲ ） 。
In the present invention, La albumen (gene numbering NM003142, inquiry derives from http://www.ncbi.nlm.nih.gov/nuccore/NM_003142) closely related with HBV DNA replication dna, experiment confirms sudden change or the gene silencing in La protein part site, the combination of La albumen and HBV RNA is produced to great effect, HBV is more easily degraded.
The present invention is according to people's hepatic pathology histogenic immunity group the selection result, it is phosphorylation La high expression level in hepatitis B tissue, the result of the low expression of phosphorylation La in normal liver tissue, the screening of employing computer virtual, by the SARS drug design based on La protein structure (http://www.pdb.org/pdb/explore/explore.do structureId=2VOD) and the method for virtual screening, carry out virtual screening through Glide5.5 molecular docking software, by multifactor scoring, filtering out 26 lead compounds has and the lead compound of the potential bonding force of people La, carry out again scoring functions scoring, flexibility analysis, transform possibility the future of structural formula, to HBV DNA, HBeAg, the inhibiting rate of HBsAg, and comprehensively analyze with the binding ability of La albumen, acquisition has the lead compound HBSC11 of pharmacologically active, 15, 34.
In the present invention, described lead compound HBSC11,15,34 can make by following synthesis path:
(1) HBSC11(chemical name: pyrazoles [1,5-a] pyridine-2-methyl-formiate, methyl pyrazolo[1,5-a] pyridine-2-carboxylate):
(2) HBSC15(chemical name: 1-cyclopropyl-1H-imidazo[4,5-c, 1-cyclopropyl-1H-imidazo[4,5-c] pyridin-2 (3H)-one):
(3) HBSC34(chemical name: (S)-2-carbobenzoxy-(Cbz)-2-pentamethylene ammonia-1-butyric acid-2-(1-methoxyl group (methyl)) acid amides, (S)-benzyl cyclopentyl (1-(methoxy (methyl) amino)-1-oxobutan-2-yl) carbamate):
Lead compound HBSC11 of the present invention, 15,34 confirms through experiment, there is in vitro more significant anti-hepatitis B virus activities, especially pyrido pyrazoles lead compound HBSC11, HBV DNA and HBeAg that people HepG2.2.15 cell strain is expressed have stronger restraining effect; Western blotting experimental result shows, compound H BSC-11 can suppress La protein expression, result shows, lead compound HBSC11 of the present invention, 15,34 is lead compound HBSC11(pyrido pyrazole compound especially) further composition optimizes, prepare preparation or the medicine of anti-hepatitis B virus activities.
The advantage of the lead compound of target human La protein of the present invention has:
1, there is the effect of stronger antiviral activity and target La albumen;
2, described lead compound and derivative thereof can be used for preparing preparation or the medicine of anti-hepatitis B virus activities or treatment hepatitis B;
3, and for the preparation of the medicine take La albumen as target.
For the ease of understanding, below the drawings and Examples by concrete are described in detail the lead compound of target human La protein of the present invention.It needs to be noted, specific examples and accompanying drawing are only in order to illustrate, obviously those of ordinary skill in the art can, according to explanation herein, make various corrections and change to the present invention within the scope of the invention, and these corrections and change are also included in scope of the present invention.
Accompanying drawing explanation
Fig. 1 has shown 26 lead compounds that virtual screening of the present invention draws restraining effect to HBV DNA in HepG2.2.15 cell, wherein, HBSC-4,11,13,14,15,16,17,19,21,22,25,12 compounds virus inhibitions of 34 are better.
Fig. 2 has shown 26 lead compounds that virtual screening of the present invention draws restraining effect to HBeAg in HepG2.2.15 cell, and wherein, HBSC-1,5,11,15,27, six compounds of 34 are better to HBeAg inhibition.
Fig. 3 has shown the impact on HepG2.2.15 emiocytosis HBsAg of 26 candidate compounds that virtual screening of the present invention draws, wherein, HBSC-1,4,11,13,15,19,20,22,25,26,28,31,13 compounds of 34 are better to HBeAg inhibition.
Fig. 4 has shown the restraining effect of lead compound of the present invention to La albumen in HepG2.2.15 cell, and wherein, HBSC11 obviously lowers La expressing quantity in cell.
Fig. 5 is the chemical structural formula of lead compound of the present invention, and wherein, carbon atom is light grey, and hydrogen atom is white, and Sauerstoffatom is grey, and nitrogen-atoms is dark, and compound is shown as club form.
Fig. 6 has shown that La protein binding site comprises Gln20, Tyr23, Arg57, Leu124, Asn139 and Ile140, wherein, two hydrogen atoms of HBSC11 are combined with the main chain of Asn139 and Ile140, two hydrogen atoms of HBSC15 are combined with the nitrogen-atoms of Gln20 and Arg57 side chain, and a hydrogen atom of HBSC34 is combined with a nitrogen-atoms of Arg57 side chain.
Embodiment
1, screening process
(1) albumen is prepared
According to La albumen and RNA compound crystal structure (PDB numbers 2VOD, downloads and obtains from albumen database Protein Data Bank), use Pymol software to delete the water molecules in crystalline structure.Delete one of them monomer and the RNA with its combination, retain the RNA of another monomer and combination thereof, and only left the innermost uridylate of RNA binding pocket (U), delete other Nucleotide.Then apply Maestro v7.5 software (Schrodinger, Inc.) " albumen preparation " module in completes a series of albumen preparation work, comprises hydrogenation, repairs the residue and the construction minimizes (minimizing the structure) etc. that lack.Finally choose the uridylate staying as " with reference to part " (reference ligand), utilize " the receptor grid generation " program in Maestro v7.5 software to generate docking pocket file, lattice (grid) size is 15.
(2) part is prepared
From SPECS chemline (network address: http://www.specs.net/), download the three-dimensional structure that obtains about 280000 compounds, about 5000 in-house compounds and SPECS chemline are integrated together to about 330000 compound structures after finally utilizing " part preparation " module in Maestro v7.5 software to be optimized.
(3) molecular docking
Utilize the Glide module of Maestro v7.5 software to carry out molecular docking.First select SP(Standard-Precision) precision, go to dock the La albumen that defines docking pocket with 330000 compound structures after optimization, according to the marking mechanism of Glide SP, obtain the compound of marking front 10000.And then select XP(Extra-Precision) precision docks this 10000 compounds, obtains approximately 9900 compounds.Afterwards, the compound that SP and XP are obtained carries out cluster, by naked eyes select (visual inspection) 26 compounds for next step active testing, wherein 10 is to buy and obtain from SPECS company, another 16 compounds come from our in-house compound library.Finally obtain 3 activated compounds, wherein HBSC-11 activity is best.
2. 26 of virtual screening the impacts of lead compound on HBV DNA replication dna
26 lead compounds are intervened to HepG2.2.15 cell 24h with 50 μ M concentration, adopt fluorescence quantitative PCR method to detect HBV DNA level in cell conditioned medium liquid.
As shown in Figure 1, the restraining effect of 26 lead compounds that virtual screening draws to HBV DNA in HepG2.2.15 cell, wherein, HBSC-4,11,13,14,15,16,17,19,21,22,25,12 compounds virus inhibitions of 34 are better, have significant difference compared with blank group.
3. 26 of virtual screening impacts that lead compound is expressed HBeAg
26 lead compounds are intervened to HepG2.2.15 cell 24h with 50 μ M concentration, adopt HBeAg level in electrochemiluminescence technology for detection cell conditioned medium liquid.
As shown in Figure 2, the restraining effect of 26 lead compounds that virtual screening draws to HBeAg in HepG2.2.15 cell, wherein, HBSC-1,5,11,15,27, six compounds of 34 are better to HBeAg inhibition, have significant difference compared with blank group.
4. 26 of virtual screening impacts that lead compound is expressed HBsAg
As shown in Figure 3, the impact of 26 candidate compounds that virtual screening draws on HepG2.2.15 emiocytosis HBsAg, wherein, HBSC-1,4,11,13,15,19,20,22,25,26,28,31,13 compounds of 34 are better to HBeAg inhibition, have significant difference compared with blank group.
5. the impact that 3 lead compounds of biological activity checking are expressed human La protein
3 lead compounds are intervened to HepG2.2.15 cell 24h with 50 μ M concentration, extract nucleoprotein, adopt La expressing quantity in immune marking technology for detection nucleus.
As shown in Figure 4, the result of comprehensive above-mentioned experiment shows, the restraining effect of 3 described compounds to La albumen in HepG2.2.15 cell, and wherein, HBSC11 obviously lowers La expressing quantity in cell, has significant difference compared with blank group.
6. application Glide5.5 molecular docking software is marked
Table 1
| Compound title | Score value docking scoring |
| HBSC11 | -5.8761 |
| HBSC15 | -5.649695 |
| HBSC34 | -3.739039 |
Result shows, Docking Score is less, and lead compound and La protein binding ability are stronger.
Result confirms, lead compound HBSC11 of the present invention, 15,34 has more significant anti-hepatitis b activity in vitro, especially pyrido pyrazoles lead compound HBSC11, HBV DNA and HBeAg that people HepG2.2.15 cell strain is expressed have stronger restraining effect, in western blotting experiment, HBSC-11 can suppress La protein expression, binding compounds molecular structural formula is combined scoring and the structural analysis of situation with La, show that lead compound of the present invention can carry out further composition optimizes; Described lead compound and derivative thereof can be used for preparing the medicine of resistance of hepatitis B, also can be used for the medicine of preparation take La albumen as target.
Lead compound HBSC11 of the present invention, 15,34 synthesis path
(1) chemical name of HBSC11 is pyrazoles [1,5-a] pyridine-2-methyl-formiate, and its synthesis path is:
(2) chemical name of HBSC15 is 1-cyclopropyl-1H-imidazo[4,5-c, and its synthesis path is:
(3) chemical name of HBSC34 is (S)-2-carbobenzoxy-(Cbz)-2-pentamethylene ammonia-1-butyric acid-2-(1-methoxyl group (methyl)) acid amides, and its synthesis path is:
Claims (1)
1. the compound H BSC11 of formula I structure is in the purposes of preparing in anti-hepatic-B virus medicine,
Described compound H BSC11 chemistry is by name: pyrazolo [1,5-a] pyridine-2-methyl-formiate.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410066584.2A CN103860538B (en) | 2011-03-23 | 2011-03-23 | The lead compound of targeting human La protein and the purposes in the medicine preparing anti-hepatitis B virus thereof |
| CN201410065464.0A CN103893173B (en) | 2011-03-23 | 2011-03-23 | The lead compound of targeting human La protein and the purposes in the medicine preparing anti-hepatitis B virus thereof |
| CN201110071212.5A CN102690262B (en) | 2011-03-23 | 2011-03-23 | Lead compound of targeting human La protein and purpose thereof in preparation of medicament for resisting hepatitis B virus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110071212.5A CN102690262B (en) | 2011-03-23 | 2011-03-23 | Lead compound of targeting human La protein and purpose thereof in preparation of medicament for resisting hepatitis B virus |
Related Child Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410066584.2A Division CN103860538B (en) | 2011-03-23 | 2011-03-23 | The lead compound of targeting human La protein and the purposes in the medicine preparing anti-hepatitis B virus thereof |
| CN201410065464.0A Division CN103893173B (en) | 2011-03-23 | 2011-03-23 | The lead compound of targeting human La protein and the purposes in the medicine preparing anti-hepatitis B virus thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102690262A CN102690262A (en) | 2012-09-26 |
| CN102690262B true CN102690262B (en) | 2014-05-28 |
Family
ID=46856014
Family Applications (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110071212.5A Expired - Fee Related CN102690262B (en) | 2011-03-23 | 2011-03-23 | Lead compound of targeting human La protein and purpose thereof in preparation of medicament for resisting hepatitis B virus |
| CN201410066584.2A Expired - Fee Related CN103860538B (en) | 2011-03-23 | 2011-03-23 | The lead compound of targeting human La protein and the purposes in the medicine preparing anti-hepatitis B virus thereof |
| CN201410065464.0A Expired - Fee Related CN103893173B (en) | 2011-03-23 | 2011-03-23 | The lead compound of targeting human La protein and the purposes in the medicine preparing anti-hepatitis B virus thereof |
Family Applications After (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410066584.2A Expired - Fee Related CN103860538B (en) | 2011-03-23 | 2011-03-23 | The lead compound of targeting human La protein and the purposes in the medicine preparing anti-hepatitis B virus thereof |
| CN201410065464.0A Expired - Fee Related CN103893173B (en) | 2011-03-23 | 2011-03-23 | The lead compound of targeting human La protein and the purposes in the medicine preparing anti-hepatitis B virus thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (3) | CN102690262B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107778310A (en) * | 2016-08-31 | 2018-03-09 | 上海市第人民医院 | Hepatitis B targets the inhibitor HBSC11 and its derivative and purposes of human La protein |
| CN113223607B (en) * | 2021-05-28 | 2023-10-20 | 北京化工大学 | Method for randomly generating heparin analogue structural coordinates in batch by adopting smiles algorithm |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1436079A (en) * | 2000-06-13 | 2003-08-13 | 布里斯托尔-迈尔斯斯奎布公司 | Imidazopyridine and imidazopyrimidine antiviral agent |
-
2011
- 2011-03-23 CN CN201110071212.5A patent/CN102690262B/en not_active Expired - Fee Related
- 2011-03-23 CN CN201410066584.2A patent/CN103860538B/en not_active Expired - Fee Related
- 2011-03-23 CN CN201410065464.0A patent/CN103893173B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1436079A (en) * | 2000-06-13 | 2003-08-13 | 布里斯托尔-迈尔斯斯奎布公司 | Imidazopyridine and imidazopyrimidine antiviral agent |
Non-Patent Citations (3)
| Title |
|---|
| La蛋白干预HBVRNA代谢的研究进展;汤静等;《中国医院药学杂志》;20090330;第29卷(第6期);第490-492页 * |
| Tian Guanghui et al.,.Synthesis and Evaluation of PDE5 Inhibitory Activity of Novel Pyrido [2＇, 1＇∶5, 1] pyrazolo [4, 3-d] pyrimidin-4-one Derivatives.《Chinese Journal of Chemistry》.2007,第25卷(第2期),第242页Scheme 1. * |
| 汤静等.La蛋白干预HBVRNA代谢的研究进展.《中国医院药学杂志》.2009,第29卷(第6期),第490-492页. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102690262A (en) | 2012-09-26 |
| CN103893173B (en) | 2016-02-24 |
| CN103893173A (en) | 2014-07-02 |
| CN103860538A (en) | 2014-06-18 |
| CN103860538B (en) | 2016-03-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Saenger | Principles of nucleic acid structure | |
| Lech et al. | Guanine base stacking in G-quadruplex nucleic acids | |
| Paulsen et al. | Inhibitor-induced structural change in the HCV IRES domain IIa RNA | |
| Virtanen et al. | Case-specific performance of MM-PBSA, MM-GBSA, and SIE in virtual screening | |
| Rutenber et al. | Binding of the anticancer drug ZD1694 to E. coli thymidylate synthase: assessing specificity and affinity | |
| Wang et al. | The systematic modeling studies and free energy calculations of the phenazine compounds as anti-tuberculosis agents | |
| Meraj et al. | Molecular modeling, docking and ADMET studies towards development of novel Disopyramide analogs for potential inhibition of human voltage gated sodium channel proteins | |
| Li et al. | Drug discovery of nucleos (t) ide antiviral agents: dedicated to Prof. Dr. Erik De Clercq on occasion of his 80th birthday | |
| Kolb et al. | Discovery of kinase inhibitors by high-throughput docking and scoring based on a transferable linear interaction energy model | |
| Elkaeed et al. | New anticancer theobromine derivative targeting egfrwt and egfrt790m: Design, semi-synthesis, in silico, and in vitro anticancer studies | |
| Wang et al. | Probing the acting mode and advantages of RMC-4550 as an Src-homology 2 domain-containing protein tyrosine phosphatase (SHP2) inhibitor at molecular level through molecular docking and molecular dynamics | |
| Ponomareva et al. | Structural and energetic properties of the potential HIV-1 reverse transcriptase inhibitors d4A and d4G: A comprehensive theoretical investigation | |
| CN102690262B (en) | Lead compound of targeting human La protein and purpose thereof in preparation of medicament for resisting hepatitis B virus | |
| El-Kalyoubi et al. | Synthesis, in silico prediction and in vitro evaluation of antitumor activities of novel pyrido [2, 3-d] pyrimidine, xanthine and lumazine derivatives | |
| Manasaryan et al. | Bioinformatic analysis of the nicotinamide binding site in poly (ADP-ribose) polymerase family proteins | |
| Radhika et al. | Role of 6-Mercaptopurine in the potential therapeutic targets DNA base pairs and G-quadruplex DNA: insights from quantum chemical and molecular dynamics simulations | |
| Mohi El-Deen et al. | Novel pyridothienopyrimidine derivatives: Design, synthesis and biological evaluation as antimicrobial and anticancer agents | |
| Rocca et al. | Chromene derivatives as selective TERRA G-quadruplex RNA binders with antiproliferative properties | |
| Liu et al. | Studies of H4R antagonists using 3D-QSAR, molecular docking and molecular dynamics | |
| Maksudov et al. | Therapeutic phosphorodiamidate morpholino oligonucleotides: Physical properties, solution structures, and folding thermodynamics | |
| Aparna et al. | 3D-QSAR studies on antitubercular thymidine monophosphate kinase inhibitors based on different alignment methods | |
| Luise et al. | Structure-based design, docking and binding free energy calculations of A366 derivatives as Spindlin1 inhibitors | |
| Spunde et al. | Design and synthesis of hepatitis B virus (HBV) capsid assembly modulators and evaluation of their activity in mammalian cell model | |
| Yamashita et al. | Synthesis of Oligonucleotides Containing 2′-N-alkylaminocarbonyl-2′-amino-LNA (2′-urea-LNA) Moieties Using Post-Synthetic Modification Strategy | |
| Wei et al. | Discovery and optimizing polycyclic pyridone compounds as anti-HBV agents |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140528 Termination date: 20180323 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |