CN110327328A - Application of 25 β--23 β of the secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative in antitumor - Google Patents
Application of 25 β--23 β of the secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative in antitumor Download PDFInfo
- Publication number
- CN110327328A CN110327328A CN201910641382.9A CN201910641382A CN110327328A CN 110327328 A CN110327328 A CN 110327328A CN 201910641382 A CN201910641382 A CN 201910641382A CN 110327328 A CN110327328 A CN 110327328A
- Authority
- CN
- China
- Prior art keywords
- mibemycin
- derivative
- isobutyl acyloxy
- cell
- secondary cyclobutenyl
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
Abstract
This programme discloses the application of 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative of mibemycin derivative technical field in antitumor, and especially 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative is used to prepare the purposes of anti-liver cancer and anti-, lung cancer, colon cancer, cervical carcinoma and medicine for nasopharyngeal.It is proved through active testing, 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative has good antitumous effect.
Description
Technical field
The invention belongs to mibemycin derivative technical field, in particular to 25 β--23 β of secondary cyclobutenyl-isobutyl acyl-oxygen
Application of the base mibemycin derivative in antitumor.
Background technique
Mibemycin (Milbemycins) is that a series of very similar 16-membered ring macrolides in structure are anti-
Raw element, can be generated by several streptomycetes, have the biological activities such as strong desinsection and mite killing.First rice was found from 1967
Since your the mould chlorins compound of shellfish, other similar compounds of a large amount of structures are reported.Have multiple mibemycins in the world
Similar drug realizes commercialization, is widely used as veterinary medicine or crop protection pesticide.
According in mibemycin class compound whether have hydrogenation benzo furan structure, it can simply be divided into α-type with
Two class formation of β-type, wherein being significantly larger than β-type with the activity of α-type structure, therefore people are main to the research of such compound
α-type is concentrated on, and the application study of β-type mibemycin class compound is extremely rare.In addition, due to mibemycin class
There is stronger desinsection, mite killing, the bioactivity of expelling parasite in compound, cause currently to grind mibemycin class compound
Study carefully to apply and be concentrated mainly on the pesticide or veterinary drug of Resistant.For mibemycin class compound answering in anti-tumor aspect
With being rarely reported.
Summary of the invention
The invention is intended to provide 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative in antitumor
Using to expand 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative purposes.
Active testing proves that 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative has anti-well
Tumor effect.Therefore, the present invention provides 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivatives to prepare
Application in anti-tumor drug or composition.
Further, 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative preparation anti-liver cancer and anti-, lung cancer,
Colon cancer, the drug of cervical carcinoma and nasopharyngeal carcinoma or the application in composition.
It is good anti-to prove that 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative has through active testing
Tumor effect is especially high to liver cancer, lung cancer, colon cancer, cervical carcinoma and nasopharyngeal carcinoma activity.
Further, 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative structural formula are as follows:
Detailed description of the invention
Fig. 1 is cancer cell line primary dcreening operation figure in the embodiment of the present invention；
Fig. 2 is 503nhibiting concentration figure of 25 β--23 β of the secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative to Hep1；
Fig. 3 is 503nhibiting concentration of 25 β--23 β of the secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative to MHCC97H
Figure；
Fig. 4 is 503nhibiting concentration figure of 25 β--23 β of the secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative to A549；
Fig. 5 is 503nhibiting concentration figure of 25 β--23 β of the secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative to LOVO；
Fig. 6 is 503nhibiting concentration figure of 25 β--23 β of the secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative to CNE1；
Fig. 7 is that 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative acts on the effect after A549 cell
Fruit figure；
Fig. 8 is that 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative acts on the effect after LOVO cell
Fruit figure；
Fig. 9 is after 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative acts on MHCC97H cell
Effect picture；
Figure 10 is that 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative acts on the effect after Hep1 cell
Fruit figure；
Figure 11 is that 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative acts on the effect after CNE1 cell
Fruit figure.
Specific embodiment
It is further described below by specific embodiment:
25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative separation method, comprising the following steps:
Step 1: extracting deposit number using ethyl acetate are as follows: the secondary generation of the streptomycete FJS 31-2 of CGMCC 4.7321
It thanks to product, obtains ethyl acetate extract medicinal extract；
Depositary institution's title of streptomycete FJS 31-2: China Committee for Culture Collection of Microorganisms's common micro-organisms center,
Preservation date: on June 2nd, 2016, classification naming: streptomycete Streptomyces sp, depositary institution address: Beijing's southern exposure
The institute 3 of area North Star West Road 1.
Step 2: carrying out mixing sample to ethyl acetate extract medicinal extract using the silica gel of 80~100 mesh, then with 200~300
Purpose silica gel obtains opposed polarity section with petroleum ether-acetone (15:1~3:1) gradient elution with petroleum ether wet method dress post
Component Fr.1~Fr.6；Fr.4 chromatographs (methanol-water 70:30~90:10) gradient elution through ODS-A column, obtains 3 components
Fr.4-1, Fr.4-2 and Fr.4-3；Component Fr.4-2 is chromatographed into (chloroform-by sephadex Sephadex LH -20 column again
Methanol 1:1) separation, further by semi-preparative HPLC, with methanol-water (72:28, v/v) for mobile phase, collection retains
Time is the component of 17.990min, and recycling design obtains white amorphous powder compound: 25 β--23 β of secondary cyclobutenyl-isobutyryl
Oxygroup mibemycin derivative, 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative structural formula are as follows:With TLC detection 254nm it is ultraviolet under visible and 10% sulfuric acid ethyl alcohol
In pink after chromogenic reagent, after being unfolded using chloroform-acetone 10:1 or petroleum ether-acetone 3:1 as solvent, Rf value is
0.5。
25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative anti-tumor test
Experimental procedure:
(1) cell recovery: -80 DEG C of cancer cell line frozen are taken out, is put into rapidly in 37 DEG C of water-baths and thaws；By cell suspension
It is transferred in the 15mL centrifuge tube of the culture medium containing 2mL, 800rpm, is centrifuged 3min.Take 1mL culture medium suspend again precipitating
Cell, and be transferred in the Tissue Culture Flask of the culture medium containing 3mL and dispel uniformly, it is placed in 37 DEG C of CO2Incubator culture.
(2) cell passes on: discarding the culture medium in culture bottle, PBS liquid cleans 2 times, and 0.25% trypsase of 1mL is added
Cellular morphology is observed under digestion inverted microscope, is terminated and is digested with 3mL complete medium when contraction is rounded.Gently dispel cell
Cell suspension is transferred to 15mL centrifuge tube by the attached cell in culture bottle, and 800rpm is centrifuged 3min.1mL is taken to cultivate base weight
The cell of the new precipitating that suspends, and be transferred in the Tissue Culture Flask of the culture medium containing 3mL and dispel uniformly, it is placed in 37 DEG C, 5%CO2Training
It supports and is cultivated in case.
(3) cell administration: taking out the cell of passage, discards the culture medium in culture bottle, and PBS is cleaned 2 times, and 1mL is added
0.25% trypsin digestion, when cellular contraction is rounded, 3mL culture medium terminates digestion.Cell is gently blown and beaten, cell is trained
The attached cell supported in bottle sufficiently falls off, and collects cell suspension and is transferred to 15mL centrifuge tube, 800rpm is centrifuged 3min.Cell is heavy
It is 5 × 10 that shallow lake culture medium, which adjusts cell concentration,4A/mL takes 100 μ L cell suspensions, is separately added into 96 orifice plate culture holes, sets
In 37 DEG C of CO2Incubator culture is for 24 hours.When cell grows to the 80% of culture hole, the drug hole is successively diluted to 2 μM, 4
μM, 8 μM, 16 μM, 32 μM, 64 μM, 128 μM, 8 concentration such as 256 μM as test concentrations, each medicine group it is parallel 6 it is multiple
Hole, 37 DEG C, 5%CO2Incubator effect terminates to cultivate for 24 hours afterwards.
(4) absorbance measurement: discarding the liquid in 96 orifice plates, and PBS cleans each orifice plate to no compound and remains, and is added 90
μ L culture medium and 10 μ L cck8 reagents, 37 DEG C are continued to terminate culture after being incubated for 4h, and microplate reader measures OD value at 490nm.
(5) experimental analysis:
According to each group OD490The average and standard deviation of value can calculate place's inhibitory rate of cell growth as follows:
IC is calculated using SPSS18.0 and GraphPadPrism6.0 statistical software50。
IC50(half maximal inhibitory concentration) refers to the 503nhibiting concentration of drug.It can refer to
Show that a certain drug or substance (inhibitor) are inhibiting certain biological process, concentration or dosage when such as cell death half,
IC50Value can be used to measure the ability of drug-induced apoptosis, i.e. inducibility is stronger, and the numerical value is lower, also can be reversed explanation
Tolerance degree of certain cell to drug.
Above-mentioned cancer cell line is thin for liver cancer cells (MHCC97H, height transfer), liver cancer cells (HEP1, low transfer), lung cancer
Born of the same parents (A549), colon cancer cell (LOVO), cervical cancer cell (Hela) or nasopharyngeal carcinoma cell (CNE1).Liver cancer cells, lung cancer are thin
Born of the same parents, colon cancer cell, cervical cancer cell are identical with test procedure with the incubation of nasopharyngeal carcinoma cell, and difference is only that culture
In the process: the DMEM basal medium of MHCC97H, the MEM basal medium of Hep1, remaining 1640 basis used
Culture medium；Above-mentioned culture medium is that 10% serum and 1% dual anti-is added in basal medium.25 β--23 β of secondary cyclobutenyl -
Isobutyl acyloxy mibemycin derivative to the 503nhibiting concentration figure of each cancer cell as figures 2-6.
The following are 25 β of various concentration--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivatives to cancer cell
Inhibiting rate situation.
25 β of various concentration--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative
To liver cancer (HEP1) cell inhibitory rate such as following table
25 β of various concentration--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative
To liver cancer (MHCC97H) cell inhibitory rate such as following table
25 β of various concentration--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative
To lung cancer (A549) cell inhibitory rate such as following table
25 β of various concentration--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative is to colon cancer (LOVO)
Cell inhibitory rate such as following table
25 β of various concentration--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative is to nasopharyngeal carcinoma
(CNE1) cell inhibitory rate such as following table
This experiment takes 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative of various concentration to cancer
Cell strain is tested through the above steps, and 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative is to difference
For the function and effect figure of cancer cell as shown in Fig. 7~11,50 μM in Fig. 7~11 in white portion are scale, indicate cell amplification
Effect picture after 200 times；In Fig. 7~8, from top to bottom, first row from left to right indicates 25 β used-secondary butylene to nine grids
- 23 β of base-isobutyl acyloxy mibemycin derivatives concentration is respectively as follows: 0 μM, 2 μM, 4 μM；Second row from left to right indicates
25 β used--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivatives concentration is respectively as follows: 8 μM, 16 μM, 32 μM；The
Three rows from left to right indicate that 25 β used--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivatives concentration is respectively as follows:
64 μM, 128 μM and 256 μM.In Fig. 9~11,25 β used--23 β of secondary cyclobutenyl-isobutyl acyloxy rice is indicated from left to right
You are respectively as follows: 0 μM, 40 μM at shellfish adm derivative concentration.
From Fig. 7~8 as can be seen that when drug concentration is at 2 μM, 4 μM, in cellular morphology and quantity simultaneously with control group
No significant difference；But when drug concentration is at 8 μM, cell is not only quantitatively reduced, but also form changes, portion
Divide cell disruption；When drug concentration increases to 16 μM, cellular morphology contraction, which is rounded, to be increased, and cell disruption is more obvious, cell
Quantity significantly reduces；With the increasing of dosage, cell death number increases, most of thin when drug concentration increases to 32 μM
Born of the same parents' floating, number cell death big absolutely.
From Fig. 9~11, it is apparent that 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivatives concentration
When reaching 40 μM, most cells floating, number cell death big absolutely.
From function and effect, it is apparent that 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative pair
Liver cancer cells, lung carcinoma cell, colon cancer cell and nasopharyngeal carcinoma cell have significant inhibiting effect；It can be widely used for preparing
In anti-tumor drug.
Claims (3)
- Application of 1.25 β--23 β of the secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative in antitumor, it is characterised in that: 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative is preparing answering in anti-tumor drug or composition With.
- 2. 25 β according to claim 1-- 23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative is antitumor In application, it is characterised in that: the tumour be liver cancer, lung cancer, colon cancer, cervical carcinoma or nasopharyngeal carcinoma.
- 3. 25 β according to claim 1 or 2-- 23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative is anti-swollen Application in tumor, it is characterised in that: 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative structural formula Are as follows:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910641382.9A CN110327328B (en) | 2019-07-16 | 2019-07-16 | Application of 25 beta-sec butenyl-23 beta-isobutyryloxymilbemycin derivative in resisting tumors |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910641382.9A CN110327328B (en) | 2019-07-16 | 2019-07-16 | Application of 25 beta-sec butenyl-23 beta-isobutyryloxymilbemycin derivative in resisting tumors |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110327328A true CN110327328A (en) | 2019-10-15 |
| CN110327328B CN110327328B (en) | 2022-07-12 |
Family
ID=68145356
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910641382.9A Active CN110327328B (en) | 2019-07-16 | 2019-07-16 | Application of 25 beta-sec butenyl-23 beta-isobutyryloxymilbemycin derivative in resisting tumors |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110327328B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110721180A (en) * | 2019-11-13 | 2020-01-24 | 遵义市第一人民医院 | Application of milbemycins compound in preparation of drugs or compositions for reversing drug resistance of cancer cells |
| CN116589606A (en) * | 2022-12-01 | 2023-08-15 | 中国药科大学 | Butyrylated yeast glucan as well as preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999017760A2 (en) * | 1997-10-02 | 1999-04-15 | Microcide Pharmaceuticals, Inc. | Fungal or mammalian cell efflux pump inhibitors for enhancing susceptibility of the cell to a drug |
| WO2016076359A1 (en) * | 2014-11-11 | 2016-05-19 | 国立研究開発法人産業技術総合研究所 | Anticancer agent containing ivermectin or milbemycin d as active ingredient |
-
2019
- 2019-07-16 CN CN201910641382.9A patent/CN110327328B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999017760A2 (en) * | 1997-10-02 | 1999-04-15 | Microcide Pharmaceuticals, Inc. | Fungal or mammalian cell efflux pump inhibitors for enhancing susceptibility of the cell to a drug |
| WO2016076359A1 (en) * | 2014-11-11 | 2016-05-19 | 国立研究開発法人産業技術総合研究所 | Anticancer agent containing ivermectin or milbemycin d as active ingredient |
Non-Patent Citations (2)
| Title |
|---|
| WENSHENG XIANG等: "Reversal of P-glycoprotein-mediated multidrug resistance in vitro by milbemycin compounds in adriamycin-resistant human breast carcinoma (MCF-7/adr) cells", 《TOXICOLOGY IN VITRO》 * |
| XIAO-QIAN LI等: "A milbemycin compound isolated from Streptomyces Sp. FJS31-2 with cytotoxicity and reversal of cisplatin resistance activity in A549/DDP cells", 《BIOMEDICINE & PHARMACOTHERAPY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110721180A (en) * | 2019-11-13 | 2020-01-24 | 遵义市第一人民医院 | Application of milbemycins compound in preparation of drugs or compositions for reversing drug resistance of cancer cells |
| CN116589606A (en) * | 2022-12-01 | 2023-08-15 | 中国药科大学 | Butyrylated yeast glucan as well as preparation method and application thereof |
| CN116589606B (en) * | 2022-12-01 | 2024-05-28 | 中国药科大学 | Butyrylated yeast glucan as well as preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110327328B (en) | 2022-07-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107298671B (en) | Selenolonic acid H from penicillium oxalicum and application thereof in preparing medicine for resisting human colon cancer | |
| CN101760496A (en) | Biological dehydrogenation preparation method of steroid drug intermediate | |
| CN107353274B (en) | Selenolonic acid I from penicillium oxalicum and application thereof in preparation of human esophageal cancer resistant medicine | |
| CN107298672B (en) | Application of seclenic acid I derived from penicillium oxalicum in preparation of anti-human colon cancer drugs | |
| CN106085868B (en) | One plant of Aspergillus and its application | |
| CN110327328A (en) | Application of 25 β--23 β of the secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative in antitumor | |
| CN103254259A (en) | Iridoid glycoside compound as well as preparation method and application thereof | |
| CN107298670B (en) | Application of medicine derived from penicillium oxalicum seclenum ketonic acid H in preparation of anti-human oral epidermoid carcinoma medicines | |
| CN107485607B (en) | Application of seclenic acid H derived from penicillium oxalicum in preparation of human esophageal cancer resistant medicine | |
| CN112300243B (en) | Cyclopeptide compound and preparation method and application thereof | |
| CN107298669B (en) | Selenolonic acid I from penicillium oxalicum and application of medicine for resisting human oral epidermoid carcinoma | |
| CN112358516A (en) | Application of diosmetin (4-O-methyl) glucoside compound in preparation of lipid-lowering drugs | |
| CN109134416B (en) | Application of seclenic acid H derived from penicillium oxalicum in preparation of human cervical cancer drugs | |
| CN109678917A (en) | Amy gram strangles mycin and its preparation method and application | |
| CN110526925A (en) | A kind of separation method of 25 β--23 β of secondary cyclobutenyl-isobutyl acyloxy mibemycin derivative and its application | |
| CN103641791B (en) | Cyclopeptide compound clavatustide B, and preparation method and application thereof | |
| CN103275139B (en) | 16 carbon diacetyl one double bond lactone type sophorolipid and application thereof | |
| CN107513072B (en) | Polyketone is inhibiting the application in HIV | |
| CN105112322A (en) | Grisic quinone A, grisic quinone B, and preparation method and medical application of grisic quinone A and grisic quinone B | |
| CN109776478B (en) | iso-Penicillium xanthone A from penicillium oxalicum and application in cervical cancer | |
| CN110934877A (en) | Perergosterol and EGFR target antibody composition and application thereof in head and neck squamous cell carcinoma | |
| CN110923278A (en) | iso-Penicillium xanthone A from penicillium oxalicum and application in lung cancer | |
| CN109369529B (en) | Method for preparing high-purity magnoflorine by taking coptis chinensis extraction mother liquor as raw material, high-purity magnoflorine prepared by method and application | |
| CN109652468B (en) | Preparation method and application of anti-tumor compound | |
| CN106399182B (en) | Algeria AUH-JLD49s and application thereof in 3' -demethyl-arctigenin conversion |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |