CN102885864A - Application of Chinese asafoetida seed extract to preparation of antitumor medicament - Google Patents
Application of Chinese asafoetida seed extract to preparation of antitumor medicament Download PDFInfo
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- CN102885864A CN102885864A CN201110204152XA CN201110204152A CN102885864A CN 102885864 A CN102885864 A CN 102885864A CN 201110204152X A CN201110204152X A CN 201110204152XA CN 201110204152 A CN201110204152 A CN 201110204152A CN 102885864 A CN102885864 A CN 102885864A
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Abstract
The invention relates to an application of a Chinese asafoetida seed extract to preparation of an antitumor medicament. The invention relates to a fat-soluble extract of Chinese asafoetida seeds and an application of the fat-soluble extract to product with an anti-tumor function. Chinese asafoetida seeds are extracted by using a steam distillation method, a supercritical technology or a fat-soluble solvent, and are analyzed by using a gas chromatography-mass spectrometry technology. The Chinese asafoetida seed extract has strong restraining effects on colon cancer, cervical cancer and hepatoma cells. The anti-tumor medicament provided by the invention can be an injection, a capsule, a tablet, a particle or in other dosage forms. The asafoetida seeds can be seeds of ferula sinkiangensis, fukangensis or ferula ovina. An anti-tumor preparation prepared from the Chinese asafoetida seed extract is mainly applied to treatment of tumor patients.
Description
Technical field
The invention belongs to medical technical field, be specifically related to Resina Ferulae seed extract anti-tumor activity and preparation method thereof and in the purposes of health food, pharmaceutical preparations.
Technical background
According to 1997 years of World Health Organization (WHO), in the whole world 5,800,000,000 populations in 1996 because of 6,300,000 people that have of cancer mortality, account for 12% of total death toll, wherein nearly 60% is pulmonary carcinoma, gastric cancer, breast carcinoma, colorectal carcinoma, oral cancer, hepatocarcinoma, cervical cancer and the esophageal carcinoma, is the 2nd large cause of the death that is only second to cardiovascular disease.The annual tumor patient of newly making a definite diagnosis in the whole world is all more than 1,030 ten thousand since 1996, to the end of the year 1999 whole world tumor patient sum exceeded 4,000 ten thousand people.World Health Organization's calendar year 2001 report, world's cancer morbidity and mortality rate have risen 22% than the nineteen ninety, also will rise about 50% in 20 years from now on.Seeking antitumor medicine from natural product has realistic meaning.
Resina Ferulae is Umbelliferae (Umbelliferae) Ferula plant, is one of long Chinese medicine of China's people's applicating history.Pharmacopoeia of People's Republic of China carries the resin [1] that the Resina Ferulae that records is Ferula sinkiangensisK.M.Shen Ferula sinkiangensis K.M.Shen or Ferula fukanensis K.M.Shen Ferula fukangensis K.M.Shen.The oleo-gum-resin with special Herba Alii fistulosi Bulbus Allii sample stink of Ferula plants secretion, have preventing the attack (or recurrence) of malaria, dysentery relieving, detoxifcation, deodorization, parasite killing, removing food stagnancy, eliminate the phlegm, the effect such as dispelling wind, among the peoplely in Uygur, Kazak etc. be usually used in treating the diseases such as meat stagnation, blood stasis mass in the abdomen, abdominal pain due to worm stagnation, have high medical value.To the research many foreign of Resina Ferulae, in recent years owing to find wherein to have phytoestrogen active component and cancer-resisting substance extremely people pay close attention to [2].Seed is as the position that contains Chuck Steak in the plant, and the research of its pharmacological action be there is not yet report.
The present invention relates to have the Resina Ferulae seed extract of function of tumor and had application in the antitumor product in preparation, filled up the blank that Resina Ferulae is used in this field.
Summary of the invention
1. the preparation method of Resina Ferulae seed extract of the present invention
(1) steam distillation: take by weighing Resina Ferulae seed g, pulverize, join in the alembic, add entry 8-12 and doubly measure, volatile oil extractor is installed, vapor distillation 3-5 hour, from volatile oil extractor is heavy, with in the moisture volatile oil separatory funnel, tells and reply a part ofly, put into-10～-15 ℃ of refrigerator and cooled and freeze and spend the night, the water intaking deicing obtains light yellow volatile oil, yield %.
(2) take by weighing the Resina Ferulae seed, pulverize, place the 5L supercritical extraction reactor.Open extraction kettle and separating still heating apparatus, open compressor pump after reaching design temperature, make extraction kettle pressure remain 20.0MPa, temperature is 50 ℃, with the CO2 flow velocity cycling extraction of 40kg/h.The separating still pressure and temperature is respectively 10.0MPa and 30 ℃, and cycling extraction 3 hours obtains yellow oil, and peat-reek is arranged, and extraction ratio is %.
(3) get Ferula sinkiangensisK.M.Shen seed 50g with petroleum ether (30-60 ℃) lixiviate of 8 times of amounts three times, each 12 hours, concentrating under reduced pressure got yellow volatile oil.
2. the chemical constituent of Resina Ferulae seed extract of the present invention
Gained Resina Ferulae extract is analyzed through GC-MS, and gas chromatography-mass spectrography technology gained mass spectrum as calculated machine mass spectrometric data library searching has been determined some chemical properties in the Resina Ferulae seed extract.GC conditions: quartz capillary column Varian VF-5MSCP8944,30m * 0.25mm, thickness 0.25 μ m.Heating schedule: since 60 ℃, be raised to 250 ℃ with 5 ℃/min, be raised to 280 ℃ with 15 ℃/min again, carrier gas is He, post flow 1.0mL/min, 270 ℃ of injector temperatures.Mass spectrum condition: EI power supply; Ionization voltage 70eV; 230 ℃ of ion source temperatures; Sweep limits 30～550aum; Sample size 1 μ l, split ratio: 22: 1.The Analysis and Identification structure shows, Resina Ferulae seed extract main chemical compositions is etc., and total content accounts for full oil.
The experiment proved that the Ferula sinkiangensisK.M.Shen seed extract has the broad-spectrum anti-tumor effect, colon adenocarcinoma cell CACO2, human cervical carcinoma cell HeLa and hepatoma carcinoma cell are all had certain inhibition growth.。Therefore, the Ferula sinkiangensisK.M.Shen seed extract can be applicable to the industries such as food, medicine, health product, cosmetics, prepares the various forms of products with antitumor action.
The specific embodiment:
The anti-human cervical cancer cell HeLa effect of embodiment 1 Ferula sinkiangensisK.M.Shen seed extract
1.1 cell strain: human cervical carcinoma cell HeLa is preserved by the conventional liquid nitrogen of this institute.
1.2 sample is volatile oil, No. 1: Ferula sinkiangensisK.M.Shen Volatile Oil from Seed (extraction by steam distillation), No. 2: Ferula fukanensis K.M.Shen Volatile Oil from Seed (extraction by steam distillation), No. 3: sheep food Resina Ferulae Volatile Oil from Seed (extraction by steam distillation), No. 4: Ferula sinkiangensisK.M.Shen seed extract (petroleum ether (30-60) extraction).
1.3 external antitumor activity is measured
Collect well-grown tumor cell, be made into 6 * 10 with the RPMI-1640 culture medium that contains 10% hyclone
4/ ml cell suspension is inoculated in 96 orifice plates, every hole 100 μ l, 37 ℃, 5%CO
2Incubator adds medicinal liquid to be measured (medicine final concentration 1.875,3.75,7.5,15,30 μ g/ml) after cultivating 24h, and every concentration is established 3 parallel holes, establishes simultaneously positive drug contrast (cisplatin 10,1,0.1 μ g/ml) and blank.Abandon supernatant after cultivating 48h, every hole adds MTT liquid 10 μ l (5mg/ml), the preparation of RPMI-1640 culture medium) continues to cultivate 4h after, every hole adds 100 μ l acidify isopropyl alcohol cell pyrolysis liquids, incubated overnight, survey absorbance (A) value with Bio-Tek MQX200 type microplate reader under detection wavelength 540nm, reference wavelength 450nm, suppression ratio calculates: (A
Blank-A
Sample)/A
Blank* 100, adopt Graphpad Prism5 statistical software to calculate IC
50
2. result
See Table 1.
Table 1 volatile oil is to the effect of tumor cell
Embodiment 2 Ferula sinkiangensisK.M.Shen seed extract resistive connection enteraden cancerous cell CACO2 effects
2.1 cell strain: colon adenocarcinoma cell CACO2, available from Chinese Academy of Sciences's Shanghai cell bank
2.2 sample is the Resina Ferulae seed extract, No. 1: Ferula sinkiangensisK.M.Shen Volatile Oil from Seed (extraction by steam distillation), No. 2: Ferula fukanensis K.M.Shen Volatile Oil from Seed (extraction by steam distillation), No. 3: sheep food Resina Ferulae Volatile Oil from Seed (extraction by steam distillation), No. 4: Ferula sinkiangensisK.M.Shen seed extract (petroleum ether (30-60) extraction).
2.3 external antitumor activity is measured
The take the logarithm CACO2 cell of trophophase is counted after the trypsinization, and adjusting cell density is 8 * 104/mL, is inoculated in the 96 porocyte culture plates every hole 100uL, 5%CO., 95% humidity, 37 ℃ cultivates 24h.The initial concentration of getting concentration and be 250ug/mL is subjected to test solution an amount of, in 1: 10 ratio gradient dilution, respectively compound concentration be 25,2.5,0.25, the tested medicine of 0.025ug/mL.Set up 5 dosage groups (by tested drug level from high to low), establish 3 parallel holes for every group, and establish matched group and background group.After cultivating 48h, add the MTT solution 50uL of 1mg/mL in every hole, continue to hatch 4h.Suck supernatant in each hole, add DMSO 150uL, dull and stereotyped shaking table jolting 10min.The 570nm place measures each hole OD value.By the suppression ratio of following formula calculating variable concentrations CACO2 cell proliferation, and take one [logrM] (the negative logarithm of drug level) as abscissa, suppression ratio as vertical coordinate, Origin v 8.0 computed in software IC
50Value.
Suppression ratio (%)=[1-(A experimental group-A background)/(A matched group-A background)] * 100%.
2.4 experimental result
2.41 the impact that the volatile oil that adopts conventional extraction by steam distillation Ferula sinkiangensisK.M.Shen (Ferula sinkiangensis K.M.Shen) seed to obtain is bred the CACO2 growth of tumour cell of In vitro culture the results are shown in Table 2, calculates IC
5016.82 ± 1.17ug/mL.
Table 2 Ferula sinkiangensisK.M.Shen Volatile Oil from Seed is on the impact of the CACO2 growth of tumour cell propagation of In vitro culture
2.42 the impact that the volatile oil that adopts conventional extraction by steam distillation Ferula fukanensis K.M.Shen seed (Ferula fukangensis K.M.Shen) seed to obtain is bred the CACO2 growth of tumour cell of In vitro culture the results are shown in Table 3, calculates IC
5012.28 ± 1.41ug/mL.
Table 3 Ferula fukanensis K.M.Shen Volatile Oil from Seed is on the impact of the CACO2 growth of tumour cell propagation of In vitro culture
2.43 the impact that the volatile oil that adopts conventional extraction by steam distillation sheep food Resina Ferulae seed (Ferula ovina) seed to obtain is bred the CACO2 growth of tumour cell of In vitro culture the results are shown in Table 4, calculates IC
5010.75 ± 1.02ug/mL.
Table 4 sheep food Resina Ferulae Volatile Oil from Seed is on the impact of the CACO2 growth of tumour cell propagation of In vitro culture
2.44 the impact that the volatile oil that adopts petroleum ether (30-60 ℃) extraction Ferula sinkiangensisK.M.Shen (Ferula sinkiangensis K.M.Shen) seed to obtain is bred the CACO2 growth of tumour cell of In vitro culture the results are shown in Table 5, calculates IC
502.41 ± 1.62ug/mL.
Table 5 Ferula sinkiangensisK.M.Shen Volatile Oil from Seed is on the impact of the CACO2 growth of tumour cell propagation of In vitro culture
Adopt three kinds of Resina Ferulaes of extraction by steam distillation to have the inhibition tumor cell effect, the size of its antitumor intensity be followed successively by sheep food Resina Ferulae greater than Ferula fukanensis K.M.Shen greater than Ferula sinkiangensisK.M.Shen.Adopting the successful of extract by solvents to be higher than steam distillation, may be owing to adopt composition kind that solvent method proposes more than steam distillation.
By above implementation example 1-2, the broad-spectrum tumor effect that the Resina Ferulae seed extract has is described, can in food, health product, cosmetics and the medicine of preparation antitumor action, use, have important practice significance and application prospect.
Claims (5)
1. the Resina Ferulae seed extract is characterized in that this extract has antitumor action.
2. Resina Ferulae seed extract according to claim 1 is characterized in that this extract is to extract from Ferula sinkiangensisK.M.Shen, Ferula fukanensis K.M.Shen or sheep food Resina Ferulae seed.
According to claim 1 with 2 described Resina Ferulae seed extract, it is characterized in that it can being vapor distillation, also can be C0
2Supercritical extraction or fat-soluble solvent extraction obtain.
4. the antitumor action of Resina Ferulae seed extract according to claim 1 is characterized in that the tumor cell that suppresses is colon cancer cell, cervical cancer cell, hepatoma carcinoma cell.
5. according to claim 1 to 4 described Resina Ferulae seed extract, it is characterized in that this extract can use in the health product that are prepared with antitumor action and medicine.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104688790A (en) * | 2014-11-15 | 2015-06-10 | 新疆医科大学附属中医医院 | Application of polar separation parts of ferula asafetida in preparation of anti-tumor drugs |
| CN105079011A (en) * | 2014-05-21 | 2015-11-25 | 中国医学科学院药用植物研究所 | Preparation and application of anti-tumor medicament |
| CN111096988A (en) * | 2020-02-20 | 2020-05-05 | 新疆医科大学第四附属医院 | Construction method and application of GC-MC (gas chromatography-monomer casting) fingerprint spectrum of asafetida volatile oil |
-
2011
- 2011-07-21 CN CN201110204152XA patent/CN102885864A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 王月娥,等: ""新疆阿魏种子化学成分的系统预实验"", 《第六届肉苁蓉暨沙生药用植物学术研讨会论文集》 * |
| 王月娥，等: "新疆阿魏种子化学成分的研究", 《中国现代中药》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105079011A (en) * | 2014-05-21 | 2015-11-25 | 中国医学科学院药用植物研究所 | Preparation and application of anti-tumor medicament |
| CN105079011B (en) * | 2014-05-21 | 2019-07-23 | 中国医学科学院药用植物研究所 | A kind of preparation and application of the drug with antitumor action |
| CN104688790A (en) * | 2014-11-15 | 2015-06-10 | 新疆医科大学附属中医医院 | Application of polar separation parts of ferula asafetida in preparation of anti-tumor drugs |
| CN104688790B (en) * | 2014-11-15 | 2019-02-01 | 新疆医科大学附属中医医院 | Asafoetide polarity separated part application in preparation of anti-tumor drugs |
| CN111096988A (en) * | 2020-02-20 | 2020-05-05 | 新疆医科大学第四附属医院 | Construction method and application of GC-MC (gas chromatography-monomer casting) fingerprint spectrum of asafetida volatile oil |
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