CN102134225A - Tricyclic diterpene lactone and preparation method and application thereof - Google Patents
Tricyclic diterpene lactone and preparation method and application thereof Download PDFInfo
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- CN102134225A CN102134225A CN2011100096636A CN201110009663A CN102134225A CN 102134225 A CN102134225 A CN 102134225A CN 2011100096636 A CN2011100096636 A CN 2011100096636A CN 201110009663 A CN201110009663 A CN 201110009663A CN 102134225 A CN102134225 A CN 102134225A
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- ethyl acetate
- silica gel
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- sesquiterpene lactone
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- -1 diterpene lactone Chemical class 0.000 title abstract description 8
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Abstract
The invention belongs to the field of organic chemistry, and more particularly relates to tricyclic diterpene lactone, the molecular structure of which is shown as formula (I). the compound can be prepared b the following method: (1) taking Rhizoma Atractylodis Macrocephalae decoction pieces, adding a proper amount of ethyl acetate for soaking and extraction for 1-3 times at room temperature, soaking and extraction each time lasts for ten days, an ethyl acetate extracting solution is combined, ethyl acetate is recovered by decompression to obtain an extractum; (2) dissolving the extractum in the ethyl acetate, adding silica gel GF254 with the weight 2-3.5 times of that of the extractum, mixing uniformly, thoroughly volatilizing the ethyl acetate, and obtaining a sample-injected silica gel; filling blank silica gel with the weight 1-3 times of that of the sample-injected silica gel into a chromatographic column and then filling the sample-injected silica gel; conducting continuous gradient elution to an eluent formed by petroleum ether and absolute ethyl alcohol with the flow rate of 0.3 to 10mL/min, collecting crystals separated out from fraction of absolute ethyl alcohol with the volume concentration of 0-0.5 percent in the petroleum ether, and then obtaining tricyclic diterpene lactone. The compound can be used for preparing medicaments for propelling bone mesenchymal stem cells to disperse to cartilage.
Description
Technical field
The invention belongs to organic chemistry filed, be specifically related to Oxygenic heterocyclic compounds, particularly three ring sesquiterpene lactones.
Background technology
(mesenchymal stem cells is to study one of maximum, that application potential is maximum stem cell at present MSCs) to mesenchymal stem cells MSCs, is to study the ideal model of breeding with differentiation mechanism.But MSCs content is few in the marrow, and a little less than the long-term proliferation activity, amplification rate is slow, and is aging to fat gradually, can't satisfy a large amount of clinical demands.Transplant in numerous in-vitro transfections and the body and studies show that the MSCs proliferation activity of transplanting is not enough, survival volume is less, has seriously limited the applied research of MSCs.And a little less than the MSCs osteogenic activity, ossific process is slower, therefore, how to improve the skeletonization potential of MSCs, makes it efficiently to chondrocyte's differentiation, will be the key issue in the MSCs differentiation research.Some Chinese medicines and extract thereof, flavonoid compound naringin naringin, hyaluronic acid, poly-l-lysine and antler polypeptide can break up to cartilage by induced dry-cell.Publication number is that the application for a patent for invention of CN101029303 discloses the material of a kind of induced dry-cell to the cartilage differentiation, this material is the mixture of stem cell-Biodegradable material, wherein said Biodegradable material is a polylactic acid PLA, polyglycolic acid PGA, polyhydroxybutyrate PGA, poly-acid anhydrides, poly-phosphazo, polyamino acid, false polyamino acid, poe, polycarbonate, the polyester urethane, poly-di-alcohol, polyethylene oxide, poly-P-Dioxane ketone, glue, gelatin, hyaluronic acid, the glycosyl glycan, chitosan, chitin, alginates or acellular matrix, and they divide combination.These mostly are macromolecular compound, and molecular weight is big, are unfavorable for permeate through cell membranes.
The bighead atractylodes rhizome (rhizoma atractylodis macroephalae) is the dry rhizome of composite family (Compositae) the plant bighead atractylodes rhizome, and warm in nature, it is sweet, bitter to distinguish the flavor of; Ground such as main product Zhejiang, Anhui; Has tonifying spleen, beneficial stomach, eliminating dampness and medium effect.The important chemical ingredients of the bighead atractylodes rhizome comprises volatile oil, lactone compound, polysaccharide, vitamin A and amino acid etc.Wherein volatile oil is about 1.4% in bighead atractylodes rhizome rhizome, is mainly atractylone, hinesol etc., is the main position of bighead atractylodes rhizome anti-tumor activity; Soluble polysaccharide is that the bighead atractylodes rhizome is brought into play oxidation resistant main position; Lactone compound is mainly the sesquiterpene lactones compounds, has isolated atractylenolide-IV, 8-β-oxyethyl group atractylenolide II, double blank art lactone, 8 at present, 9-epoxy atractylodes lactone, 4,15 one epoxy hydroxyl atractylodes lactones etc.Wherein, atractylenolide and atractylenolide II have an anti-inflammatory action, be widely used in clinical treatment aging, struvite osteopathia, so in many treatments and marrow, generally use the bighead atractylodes rhizome and other bulk drug to carry out compatibility in traditional prescription of MSC contents level diseases associated.Yet, mesenchymal stem cells MSCs is not appeared in the newspapers to the cartilage Differentiation from isolated monomer wherein.
Summary of the invention
For solving the problems of the technologies described above, the invention provides a kind of tricyclic sesquiterpene lactone, this compound has short mesenchymal stem cells MSCs to the cartilage Differentiation.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
A kind of tricyclic sesquiterpene lactone, its molecular structure is as shown in the formula shown in the I:
The molecular formula of compound shown in the above-mentioned formula I is for being C
15H
20O
3Molecular weight is 248.0, normal temperature and pressure is colourless acicular crystal down, called after: 3,8a-dimethyl-5-methylene radical-9a-hydroxyl-perhydronaphthalene [2,3-b]-1,4-dihydrofuran-2-ketone is [English by name: 3,8a-dimethyl-9a-hydroxyl-5-methylene-decahydronaphthalene [2,3-b]-1,4-2H-furan-2-one].
Tricyclic sesquiterpene lactone of the present invention can obtain by the method for chemosynthesis, also can separate obtaining from the Chinese medicine bighead atractylodes rhizome, and wherein, the step of described chemical synthesis process and processing condition are knowledge well-known to those having ordinary skill in the art.The above-mentioned bighead atractylodes rhizome is that the method that the dry rhizome bighead atractylodes rhizome of feverfew bighead atractylodes rhizome Atractylodes macrocephala Koidz. therefrom separates tricyclic sesquiterpene lactone of the present invention is made up of following steps:
(1) get bighead atractylodes rhizome medicine materical crude slice, add to soak under an amount of ethyl acetate room temperature and extract 1～3 time, the each immersion extracted ten days, the combined ethyl acetate extracting solution, and the reclaim under reduced pressure ethyl acetate gets medicinal extract;
(2) gained medicinal extract is dissolved in the ethyl acetate, admixes the heavy silica GF254 of 2～3.5 times of medicinal extract, mix, volatilize ethyl acetate, obtain application of sample silica gel; The blank silica gel that the gained application of sample silica gel weight of packing in the chromatography column earlier is 1～3 times, the application of sample silica gel of packing into then; The elutriant of forming with sherwood oil and dehydrated alcohol carries out the continuous gradient wash-out with the flow velocity of 0.3～10ml/min, collects the volumetric concentration of dehydrated alcohol in sherwood oil and be the crystal of separating out in 0～0.5% the flow point, obtains the tricyclic sesquiterpene lactone.
In the above-mentioned separation method, the flow velocity of described elutriant is relevant with the particle diameter of silica gel, and the big stream of grain is hurried up, otherwise flows a little slower.
In the above-mentioned separation method, described elutriant is by sherwood oil and dehydrated alcohol two phase composites, because wherein the maximum concentration of dehydrated alcohol in sherwood oil only is 0.5%, if therefore use some simple and crude equipments, drips as bottle, just is difficult to dispensing equably.In order to address the above problem, described continuous gradient elution process can adopt following simple method: with the container A sherwood oil of packing into, another container B dress dehydrated alcohol volumetric concentration is 0.5% sherwood oil and dehydrated alcohol blended solution, then with container B, container A and chromatography column upper and lower settings successively and connection, make container B and container A begin following at last simultaneously, and in following process, the solvent in the container A is stirred.
Tricyclic sesquiterpene lactone of the present invention has short mesenchymal stem cells MSCs to the cartilage Differentiation, can be used for preparing the medicine for the treatment of cartilage injury, this medicine is made up of compound and pharmaceutical excipient shown in the formula I, can be injection, oral tablet, capsule etc.
What will further prove tricyclic sesquiterpene lactone of the present invention below by experiment has short mesenchymal stem cells MSCs to the cartilage Differentiation.
1 materials and methods
1.1 reagent and laboratory animal
Cell cultures and reagent: low sugar Da Shi revises Yi Shi substratum (low glucose dulbecco ' s modified eagle ' smedium, L-DMEM), high sugared Da Shi revise the Yi Shi substratum (high glucose dulbecco ' s modified eagle ' smedium, H-DMEM), L-DMEM contains D-glucose 1g/L and L-glutaminate; H-DMEM contains 4.5g/L D-glucose and L-glutaminate, Percoll stock solution and ox tire serum, and (fetal bovine serum FBS) is Gibcol company product; CO
2Incubator, German Heraeus company product; Inverted phase contrast microscope, Japanese Olympus company product; The CD44 monoclonal mouse antibody is provided by Shenzhen brilliant U.S. company.
Laboratory animal: purebred SD rat, the cleaning level, male and female half and half, at 1～4 month monthly age, body weight 250～300g is provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.
1.2.MSCs separate, increase and identify
Rat femur, shin bone marrow are gone out with the DMEM that contains 10%FBS, and fully mixing changes centrifuge tube over to, and the centrifugal 10min of 300g removes supernatant, and L-DMEM is resuspended, centrifugal remove supernatant after, add 4ml L-DMEM mixing.The Percoll stock solution mixed by 0.56: 0.44, got 4ml and put into the 10ml centrifuge tube, inhaled bone marrow fluid and was slowly adding from liquid level 2cm place subsides tube wall.The centrifugal 30min of horizontal centrifuge 900g, the collecting monocytic cell layer is used the DMEM washed twice.Counting cells is adjusted density then, by 1 * 10
6/ cm
2Density inoculation, 37 ℃, the CO of 5% saturated humidity
2Incubator is cultivated, and nutrient solution is the L-DMEM that contains 10%FBS, changes nutrient solution behind the 5d, discards not attached cell, and 3～4d changes liquid 1 time, with containing 0.25% pancreatin room temperature digestion, 2～3min, presses 1 * 10 near the MSC that merges
4/ cm
2Go down to posterity.After cultivating for 3 generations, take out and do the Flow Cytometry detection.
1.3.MSCs induce the cartilage differentiation culture
Pass behind the MSCs in 3 generations with 1 * 10
5/ cm
2Cell density is inoculated into 24 well culture plates and 6 orifice plates, divides negative blank group and tricyclic sesquiterpene lactone to induce group; The tricyclic sesquiterpene lactone component does not add 3,30 μ g/ml tricyclic sesquiterpene lactone of the present invention, and the blank group adds solvent and handles factor, and every 3d changes liquid 1 time, induces 7 days.
1.4 Toluidine blue staining detects cell glycosaminoglycan content
To grow to the cell of logarithmic phase by 1 * 10
5/ cm
2Density is inoculated in the 24 hole versions of cover glass, above-mentioned each group is induced 7 days respectively after, PBS rinsing cover glass, 1% Toluidine blue staining is after 10 minutes, tri-distilled water embathes, the resinene sealing, light microscopic is observed down.
1.5RT-PCR detect cartilage phenotype related protein gene, SOX9 and Shh signal path
Extract the genome of blank group, tricyclic sesquiterpene lactone group, total RNA carries out according to Trizol test kit specification sheets; RT-PCR adopts Primer Premier5.0 software design primer with reference to the cDNA sequence of Genbank, and is synthetic by Shanghai Ying Jun company.OPN upstream primer: 5-AGTTTGGCAGCTCAGAGGAGAAG-3, downstream primer: 3-GATTGGAGTCAAAACGTCTGCTTG-5; CollagenIX upstream primer: 5-TACCTGGCATGAAGGGCAGTG-3, downstream primer: GTCCTGGAATTCCTGGAGA; Epiphycan upstream primer: 5-GCAAGCTTTGTGTACTGTGA-3, downstream primer: 3-GCTCTGTAGTTGGTGCAG-5; SOX9 upstream primer: 5-CGGCGGAGGAAGTCGGTGAAGA-3, downstream primer: 3-CGTTGGGCGGCAGGTATTGGTC-5; Patched upstream primer: 5-CCTCCTTTACGGTGGACAAA-3, downstream primer: 3-CAGTGGCGTTCTTGACGG-5; Gli-1 upstream primer: 5-AAGCATCAGAACCGCACCCACTC-3, downstream primer: 3-CCACCCGTGTTGCCCGTCATCTC-5; GAPDH upstream primer: 5-TAAAGGGCATCCTGGGCTACACT-3, downstream primer: 5-TTACTCCTTGGAGGCCATGTAGG-3.Reaction conditions is: 93 ℃ of 2min, 93 ℃ of 45seconds then, 63 ℃ of 30secends, totally 30 circulations.Reaction is got the PCR reaction solution and is carried out agarose gel electrophoresis after finishing.The RT-PCR product carries out absorbancy scanning through 1.5% agarose gel electrophoresis with image analyzer.
2 results
2.1 the Flow Cytometry result utilizes Flow cytometry cell-surface antigens CD44 cell sign, found that the CD44 positive cell reaches 98.14% (see figure 1), shows that cultivating resulting cell is MSC.
2.2 Toluidine blue staining found that, tricyclic sesquiterpene lactone rising cell glycosaminoglycan content (see figure 2) shows that the tricyclic sesquiterpene lactone promotes the cartilage of inducing of MSCs to break up.
2.2 the tricyclic sesquiterpene lactone promotes the cartilage phenotype Expression of Related Genes
0,3,30 μ g/ml tricyclic sesquiterpene lactones acted on MSCs respectively 3 days, 6 days, and collecting cell extracts cell RNA, and RT-PCR detects CollagenIX, the variation of Epiphycan and OPN.The result of RT-PCR shows that the tricyclic sesquiterpene lactone promotes the expression of cartilage phenotype genes involved CollagenIX (see figure 3), Epiphycan (see figure 4), OPN (see figure 5).Fig. 3 ordinate zou is represented the ratio of CollagenIX and confidential reference items GAPDH; *, compare with control group in P＜0.05.Fig. 4 ordinate zou is represented the ratio of Epiphycan and confidential reference items GAPDH; *, compare with control group in P＜0.05.Fig. 5 ordinate zou is represented the ratio of OPN and confidential reference items GAPDH; *, compare with control group in P＜0.05.
2.3 the tricyclic sesquiterpene lactone promotes cartilage phenotype associated transcription factor Sox9 expression of gene
0,3,30 μ g/ml tricyclic sesquiterpene lactones acted on MSCs respectively 3 days, 6 days, and collecting cell extracts cell RNA, and RT-PCR detects the variation of Sox9.The tricyclic sesquiterpene lactone that shows of RT-PCR promotes cartilage phenotype associated transcription factor Sox9 expression of gene (see figure 6).Fig. 6 ordinate zou is represented the ratio of Sox9 and confidential reference items GAPDH.*, compare with control group in P＜0.05.
2.4 the tricyclic sesquiterpene lactone activates the Shh signal path
0,3,30 μ g/ml tricyclic sesquiterpene lactones acted on MSCs respectively 3 days, 6 days, and collecting cell extracts cell RNA, and RT-PCR detects the variation of patched gene and Gli-1.The result of RT-PCR shows that the tricyclic sesquiterpene lactone promotes the expression (seeing Fig. 7 and 8) of Shh signal path acceptor patched gene and Shh signal path target gene Gli-1.Fig. 7 ordinate zou is represented the ratio of patched and confidential reference items GAPDH.*, compare with control group in P＜0.05.Fig. 8 ordinate zou is represented the ratio of Gli-1 and confidential reference items GAPDH.*, compare with control group in P＜0.05.
2.5 the tricyclic sesquiterpene lactone promotes the cartilage phenotype expression of gene to depend on the Shh signal path
Tricyclic sesquiterpene lactone, Cyclopamine, tricyclic sesquiterpene lactone and Cyclopamine acted on MSCs respectively 6 days, and collecting cell extracts cell RNA, and RT-PCR detects the variation (see figure 9) of Sox9 and Gli-1.The result of RT-PCR shows that Shh signal path blocker Cyclopamine suppresses the tricyclic sesquiterpene lactone and promotes cartilage phenotype expression of gene (see figure 10).Fig. 9 ordinate zou is represented the ratio of Sox9 and confidential reference items GAPDH; #, compare with control group in P＜0.05; * compare with tricyclic sesquiterpene lactone group in P＜0.05.Figure 10 ordinate zou is represented the ratio of Gli-1 and confidential reference items GAPDH.#, compare with control group in P＜0.05; * compare with tricyclic sesquiterpene lactone group in P＜0.05.Prompting tricyclic sesquiterpene lactone promotes the cartilage phenotype expression of gene to depend on the Shh signal path.
In sum, the present invention acts on the different time of MSCs with the tricyclic sesquiterpene lactone of different concns, use the method detection cartilage phenotypic differentiation expression of gene that Toluidine blue staining detects cell glycosaminoglycan content and RT-PCR, determined the effect that the tricyclic sesquiterpene lactone induces MSCs to break up to cartilage.
Description of drawings
Fig. 1 Flow cytometry is cultivated 3 generation MSCs surface antigen CD44 marking patterns.
Fig. 2 tricyclic sesquiterpene lactone acts on 6 days Toluidine blue staining figure of MSCs respectively.
Fig. 3 tricyclic sesquiterpene lactone promotes the expression of cartilage phenotype genes involved CollagenIX.
Fig. 4 tricyclic sesquiterpene lactone promotes the expression of cartilage phenotype genes involved Epiphycan.
Fig. 5 tricyclic sesquiterpene lactone promotes the expression of cartilage phenotype genes involved OPN.
Fig. 6 tricyclic sesquiterpene lactone promotes the expression of the relevant associated transcription factor Sox9 of cartilage phenotype.
Fig. 7 tricyclic sesquiterpene lactone promotes the expression of Shh signal path acceptor patched.
Fig. 8 tricyclic sesquiterpene lactone promotes the expression of Shh signal path target gene Gli-1.
Fig. 9 Shh signal path blocker Cyclopamine suppresses the expression that the tricyclic sesquiterpene lactone promotes cartilage phenotype associated transcription factor Sox9.
Figure 10 Shh signal path blocker Cyclopamine suppresses the expression that the tricyclic sesquiterpene lactone promotes Gli-1.
Embodiment
1, the preparation of compound
(1) get 1kg bighead atractylodes rhizome medicine materical crude slice and add an amount of ethyl acetate, exceed, at room temperature soak 2 times, soaked ten days at every turn to soak fully, the combined ethyl acetate extracting solution, the reclaim under reduced pressure ethyl acetate gets medicinal extract 24g.
(2) medicinal extract is dissolved in the 48mL ethyl acetate, admixes 200～300 purpose silica GF254s of 76g, mix, volatilize ethyl acetate, obtain application of sample silica gel;
(3) in chromatography column, the blank silica gel of the 76g that packs into earlier reinstalls application of sample silica gel;
(4) concentration of packing in container A is 100% sherwood oil, and the dehydrated alcohol volumetric concentration of packing in the container B is 0.5% dehydrated alcohol and sherwood oil blended solution, and container B is suspended from the top of container A, is communicated with, and outlet is joined with the top of chromatography column under the container A; Then, solvent in container B and the container A is dripped downwards simultaneously, wherein the mixing solutions in the container B is added dropwise in the sherwood oil of container A with 0.3-10ml/min speed, and stir, solvent in the container A injects the chromatography column upper end with 0.3-10ml/min speed and carries out wash-out, collect the volumetric concentration of dehydrated alcohol in sherwood oil and be the crystal of separating out in 0～0.5% the flow point, obtain tricyclic sesquiterpene lactone 4.8g, yield is 0.06%.
2, the evaluation of compound
Identify: molecular formula C
15H
22O
3, normal temperature and pressure is colourless acicular crystal down; EI-MS (M)
+: 248.0; IR v
Max KBr(cm
-1): 2933,1759,1674,1649,889.UVλ
max?
MeOH：242nm；
1H-NMR,
13C-NMR, HMBC data see Table 1.
Table 1
1H-NMR (400MHz) and
13C-NMR (100MHz)
Embodiment 2
Prescription:
With the compound that example 1 obtains, adopt the preparation method of medical injection to make injection liquid.This compound 500mg is dissolved in 1,2 propylene glycol 20ml and tween-80 2ml, adds the injection water again to 250ml.Pour into after mixing in the bottle, sealing, the injection liquid of one of 2mg/1ml is made in sterilization.This product is used for intravenous injection (iv.), is grown up 2mg/ days, is equivalent to 1; Children's's usual amounts: by body weight 0.05mg/kg/ days.
Prescription:
The compound that example 1 is obtained adopts the preparation method of capsule for medicine to make capsule.With this compound 25000mg, add lime carbonate 4500mg, add starch 500mg.Mix and make particle, can becomes that every intragranular is tolerant to be the capsule of 30mg, and every capsules contains this product 25mg.
Instructions of taking: oral, adult's usual amounts, every day 2 times, each 1-2 grain; Children's's usual amounts, by 0.5mg/kg of body weight, per six hours once.4 weeks were a course of treatment, and after two courses of treatment, dose cuts down according to the circumstance.
Embodiment 4
Prescription:
The compound that example 1 is obtained adopts the preparation method of medicinal tablets to make tablet, with this compound 40000mg, adds lime carbonate 9000mg, adds starch 1000mg.Mix, cross 90 mesh sieves, compressing tablet.Make sheet and heavily be the tablet of 50mg, every contains this product 40mg.
Instructions of taking: oral, adult's usual amounts, every day 2 times, each 1-2 sheet; Children's's usual amounts, by 0.5mg/kg of body weight, per six hours once.4 weeks were a course of treatment, and after two courses of treatment, dose cuts down according to the circumstance.
Claims (6)
1. tricyclic sesquiterpene lactone, its molecular structure is as shown in the formula shown in the I:
2. the preparation method of the described tricyclic sesquiterpene lactone of claim 1, this method is made up of following steps:
(1) get bighead atractylodes rhizome medicine materical crude slice, add to soak under an amount of ethyl acetate room temperature and extract 1～3 time, the each immersion extracted ten days, the combined ethyl acetate extracting solution, and the reclaim under reduced pressure ethyl acetate gets medicinal extract;
(2) gained medicinal extract is dissolved in the ethyl acetate, admixes the heavy silica GF254 of 2～3.5 times of medicinal extract, mix, volatilize ethyl acetate, obtain application of sample silica gel; The blank silica gel that the gained application of sample silica gel weight of packing in the chromatography column earlier is 1～3 times, the application of sample silica gel of packing into then; The elutriant of forming with sherwood oil and dehydrated alcohol carries out the continuous gradient wash-out with the flow velocity of 0.3～10ml/min, collects the volumetric concentration of dehydrated alcohol in sherwood oil and be the crystal of separating out in 0～0.5% the flow point, obtains the tricyclic sesquiterpene lactone.
3. the preparation method of tricyclic sesquiterpene lactone according to claim 2, the method that it is characterized in that described continuous gradient wash-out is: with the container A sherwood oil of packing into, another container B dress dehydrated alcohol volumetric concentration is 0.5% sherwood oil and dehydrated alcohol blended solution, then with container B, container A and chromatography column upper and lower settings successively and connection, make container B and container A begin following at last simultaneously, and in following process, the solvent in the container A is stirred.
4. the described tricyclic sesquiterpene lactone of claim 1 is in the application of the short mesenchymal stem cells MSCs of preparation in the medicine of cartilage differentiation.
5. medicine for the treatment of cartilage injury, this medicine is made up of described derivative of claim 1 and pharmaceutical excipient.
6. a kind of medicine for the treatment of cartilage injury according to claim 5 is characterized in that this medicine is injection, oral tablet or capsule.
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| CN106190965A (en) * | 2016-07-19 | 2016-12-07 | 袁肇斌 | A kind of mesenchymal stem cells MSCs In vitro culture liquid and cultivation amplification method |
| CN107574147A (en) * | 2017-10-20 | 2018-01-12 | 南京盖斯夫医药科技有限公司 | A kind of mescenchymal stem cell Proliferation, Differentiation nutrient solution using atractylenolide as trophic factors |
| CN107674053A (en) * | 2017-10-23 | 2018-02-09 | 南京盖斯夫医药科技有限公司 | A kind of atractylodes lactone V, preparation method and the application in terms of CIK cell culture |
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| 《Chemical & Pharmaceutical Bulletin》 19791231 Katsuya endo et al Antiinflammatory principles of atractylodes rhizomes 2954-2958 1,5,6 第27卷, 第12期 * |
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| CN107674053B (en) * | 2017-10-23 | 2019-08-20 | 北京恒生佳合细胞科技有限公司 | A kind of atractylodes lactone V, preparation method and the application of aspect is frozen in CIK cell |
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| CN112939909A (en) * | 2021-01-15 | 2021-06-11 | 宁波大学 | Nano-docetaxel condensation compound and preparation method and application thereof |
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