CN112274539B - Use of bamboo-cypress seed oil for preventing or treating cancer - Google Patents

Use of bamboo-cypress seed oil for preventing or treating cancer Download PDF

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CN112274539B
CN112274539B CN202011199881.6A CN202011199881A CN112274539B CN 112274539 B CN112274539 B CN 112274539B CN 202011199881 A CN202011199881 A CN 202011199881A CN 112274539 B CN112274539 B CN 112274539B
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雍建平
卢灿忠
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Xiamen Institute of Rare Earth Materials
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/13—Coniferophyta (gymnosperms)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/31—Extraction of the material involving untreated material, e.g. fruit juice or sap obtained from fresh plants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction

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Abstract

The invention discloses a new application of bamboo-cypress seed oil in preventing or treating cancers, and the in-vitro anti-tumor test of the bamboo-cypress seed oil and the anti-cancer pharmaceutical composition thereof prepared by the invention shows that the bamboo-cypress seed oil has stronger inhibitory activity on lung adenocarcinoma cell strains (A549), gastric cancer cell strains, breast cancer cell strains (MCF-7) and cervical cancer cell strains (Hela). Can be used for preventing and treating tumor/cancer diseases.

Description

Use of bamboo-cypress seed oil for preventing or treating cancer
Technical Field
The invention belongs to the technical field of medicines. In particular to the application of the bamboo cypress seed oil in the aspect of preventing or treating cancer.
Background
Cancer has become the leading fatal disease worldwide. Cancer can occur in various organs and tissues at any age, and the main cancer types that lead to death are: lung cancer, gastric cancer, liver cancer, colon cancer, breast cancer, and the like. In recent years, lung cancer has become a malignant tumor with the highest incidence. According to the World Health Organization (WHO) statistics: the number of diagnosed lung Cancer in 2008 is about 160 million, about 13% of the total number of Cancer cases, about 140 million deaths, and about 18% of the total number of Cancer deaths (Jemal a, et al.c a Cancer j.clin.2011,61: 69-90); in 2010, 222,520 new lung Cancer patients in the United states, 157,330 deaths, of which non-small cell lung Cancer (NSCLC) patients died about 85% of the total lung Cancer deaths (Jemal A, et al.C A Cancer J.Clin.2010,60: 277-300). In 2020, the united states is expected to have newly diagnosed about 180 ten thousand cancer cases, which corresponds to about 4950 new cancer cases each day. It is expected that about 60 million people will die from cancer in the united states in 2020, which is equivalent to 1600 people die from cancer per day. The largest number of deaths occur in men from lung, prostate and colon and rectal cancers, while the largest number of deaths occur in women from lung, breast and colon and rectal cancers. Lung Cancer causes nearly one-fourth of all deaths and is the first Cancer to justify its mortality (Rebecca l. siegel, et al. Cancer staticiscs, 2020. CA: a Cancer Journal for Clinicians, 2020.).
According to the latest article data published in 2019 by the national cancer center in China (Zhengrongshou, etc., the epidemic situation of malignant tumors in China in 2015 is analyzed, and the Chinese tumor journal, 2019,41(1): 19-28): 392.9 ten thousand cases of new malignant tumors in China in 2015, the incidence rate of the malignant tumors is 285.83/10 ten thousand, the standardization rate of Chinese population is 190.64/10 ten thousand, which is higher than the standardization rate of world population 186.09/10 ten thousand; in 2015, the death rate of malignant tumors is about 233.8 ten thousand, 170.05/10 ten thousand, the winning rate of death is 106.72/10 ten thousand, and the winning rate of death is higher than the world standard of death rate 105.84/10 ten thousand. The mortality rate of malignant tumor in rural area is 166.79/10 ten thousand, and the mortality rate of winning a bid is 110.76/10 ten thousand. Common malignant tumors in China include lung cancer, gastric cancer, colorectal cancer, liver cancer, female breast cancer and the like. The first 10 cases accounted for approximately 76.70% of all new cases of malignancy. Lung cancer, liver cancer, stomach cancer, esophageal cancer, colorectal cancer and the like are the main causes of tumor death, and the first 10 deaths account for about 83.00% of all malignant tumor deaths.
However, most cancer patients find the disease in the middle to late stage, the overall effect of clinical treatment is poor, and especially the multi-drug resistance is continuously appeared, so that the treatment of the cancer is difficult. Therefore, the development of novel anticancer drugs having high activity and low side effects to meet clinical needs is urgent. In particular to a functional product which is found from medicinal and edible medicinal materials and has anti-tumor activity, which is more favored by patients.
Bamboo cypress (Podocarpus nagi), belonging to Podocarpaceae (Podocarpaceae), originated from about 1 hundred million 5500 ten thousand years ago of middle generation chalk era, is called as activated stone and is a national secondary protective plant. It is precious in whole body, has the reputation of 'plant diamond' and is a precious rare endangered tree species. The root, stem and leaf and seed of the bamboo cypress are rich in various chemical active ingredients, and the Yao nationality is used for treating fracture, incised wound, gunshot wound, lumbar muscle strain, mental disease, bromhidrosis, eye diseases, cold and the like. On the other hand, the bamboo cypress is rich in a large amount of flavone and beta-elemene, the flavone has more functions and has the effects of resisting cancers, controlling blood sugar, enhancing immunity and the like, and the beta-elemene is a new national class II anti-cancer medicine. In earlier studies, we extracted the bamboo cypress seed oil and studied its chemical composition. The oil meets the standard of edible oil, and has antioxidant activity and hypotensive activity (Yongjian, Lu and lau faithful, Zhang Shibi, Chinese patent application No. CN 202010203003.0). The oil can be used as functional oil for both medicine and food, and has good application prospect.
Disclosure of Invention
The invention aims to provide a new application of edible bamboo-cypress seed oil in the aspects of preventing and treating tumors/cancers, aiming at the current situation that clinically applied anti-tumor/cancer drugs have toxicity and great side effects.
The invention provides an application of bamboo cypress seed oil in preparing an anti-cancer drug.
According to an embodiment of the present invention, the cancer may be gastric cancer, breast cancer, cervical cancer and lung adenocarcinoma.
The invention provides an anticancer pharmaceutical composition, which comprises effective amount of bamboo-cypress seed oil.
According to the embodiment of the invention, the bamboo cypress seed oil is prepared by the following steps:
(1) crushing the bamboo cypress seeds to obtain bamboo cypress seed powder;
(2) steaming the bamboo cypress seed powder obtained in the step (1);
(3) extruding the steamed bamboo cypress seed powder obtained in the step (2) to obtain oil;
(4) standing and filtering the extruded oil obtained in the step (3) to obtain the bamboo-cypress seed oil.
According to an embodiment of the present invention, the raw material of the bamboo cypress seeds is bamboo cypress nuts (bamboo cypress seeds are peeled).
According to an embodiment of the present invention, the weight of the seeds of bamboo cypress in step (1) is 10 to 100kg, preferably 30 to 70kg, for example 50 kg.
According to an embodiment of the present invention, the particle size of the bamboo cypress seed powder in step (1) may be 20 to 70 mesh, preferably 30 to 60 mesh, for example 40 mesh.
According to an embodiment of the present invention, the steam of step (2) may be implemented by using water vapor, and the equipment of the steam may be any one of a water bath and a steamer; the steaming time may be 5 to 30 minutes, preferably 10 to 25 minutes, more preferably 18 to 22 minutes, for example, about 20 minutes.
According to the embodiment of the invention, the extrusion in the step (3) can be realized by an extrusion device, and the pressure of the extrusion can be 0-50 MPa, for example, the pressure is increased by adopting a 0-30 MPa procedure;
the extrusion time is 5 to 60 minutes, preferably 10 to 40 minutes, more preferably 20 to 30 minutes, for example 25 minutes.
Preferably, the fine powder of steamed bamboo and cypress seeds obtained in step (2) can be artificially pressed into a fixed shape, such as a cake shape, in a fixed die before being extruded.
According to an embodiment of the present invention, the standing time in step (4) is 6 to 48 hours, preferably 8 to 24 hours, for example 12 hours.
According to an embodiment of the present invention, any one of a buchner funnel and a sand core funnel, preferably a 4 μm sand core funnel, may be used for the filtration in step (4).
According to the embodiment of the invention, the content (mass percentage) of the total flavone (calculated by rutin) in the bamboo-cypress seed oil is more than or equal to 4 percent, such as 4.23 percent.
According to an embodiment of the invention, the content of unsaturated fatty acids in the bamboo cypress seed oil is 75% or more, for example 75.5%.
According to an embodiment of the invention, the content of linoleic acid in the bamboo cypress seed oil is more than or equal to 35%, for example 38.2%.
According to an embodiment of the invention, the content of arachidonic acid in the bamboo-cypress seed oil is more than or equal to 1%, for example 1.40%.
According to an embodiment of the invention, the content of arachidilic acid in the bamboo cypress seed oil is 8% or more, for example 8.04%.
According to an embodiment of the invention, the content of vitamin E in the bamboo-cypress seed oil is more than or equal to 2mg/100g, for example 2.11mg/100 g.
According to the embodiment of the invention, the clearance rate of the bamboo-cypress seed oil to oxygen free radicals is more than or equal to 86%, and preferably more than or equal to 88%.
According to an embodiment of the present invention, the pharmaceutical composition may be a solid oral formulation or an injectable formulation.
According to an embodiment of the present invention, the solid oral preparation may be a capsule, a tablet, a granule, a sustained-release agent, or the like.
According to the embodiment of the invention, the injection can be small needle injection and freeze-dried injection.
According to an embodiment of the invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients; the auxiliary materials comprise one or a mixture of a plurality of fillers, binders, disintegrating agents, lubricants, flavoring agents, enteric coatings or slow-release materials.
According to an embodiment of the invention, the filler comprises: one or more of lactose, sucrose, dextrin, starch, mannitol, sorbitol, calcium hydrogen phosphate, calcium carbonate and microcrystalline cellulose;
according to the embodiment of the invention, the adhesive comprises one or a mixture of sucrose, starch, sodium carboxymethyl cellulose, methyl cellulose, polyethylene glycol and water;
according to the embodiment of the invention, the disintegrating agent comprises one or a mixture of more of starch, cross-linked polyketone, cross-linked sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose and sodium carboxymethyl cellulose;
according to the embodiment of the invention, the lubricant comprises one or a mixture of more of talcum powder, magnesium stearate, stearic acid, superfine silica powder, polyethylene glycol-4000 and polyethylene glycol-6000;
according to the embodiment of the invention, the flavoring agent comprises one or a mixture of sucrose, sorbitol, saccharin sodium, maltitol, stevioside and aspartame;
according to the embodiment of the invention, the slow release material comprises one or a mixture of several of hypromellose and guar gum with different specification viscosities.
According to the embodiment of the invention, when the anticancer pharmaceutical composition is a solid oral preparation, the anticancer pharmaceutical composition comprises the following components in parts by weight: 0.01-0.5 part of bamboo-cypress seed oil, 2-20 parts of lactose, 1-10 parts of pregelatinized starch, 1-5 parts of microcrystalline cellulose, 0.1-5 parts of carboxymethyl starch sodium, 2-10 parts of 10% povidone solution, 1-5 parts of magnesium stearate and 1-5 parts of talcum powder.
According to an embodiment of the present invention, when the anticancer pharmaceutical composition is a solid oral preparation, the anticancer pharmaceutical composition comprises the following components in parts by weight: 0.078 parts of bamboo-cypress seed oil, 13 parts of lactose, 3.5 parts of pregelatinized starch, 2.5 parts of microcrystalline cellulose, 0.8 part of carboxymethyl starch sodium, 5 parts of 10% povidone solution, 1.5 parts of magnesium stearate and 1.5 parts of talcum powder.
According to an embodiment of the present invention, when the anticancer pharmaceutical composition is a solid oral preparation, the preparation method thereof comprises: mixing the auxiliary materials, sieving with a 40-100 mesh sieve, adding the bamboo and cypress seed oil into 10% povidone solution, mixing, adding into the sieved auxiliary materials, grinding, mixing, drying at 50-60 ℃ for 2-3 hours, and tabletting to obtain the anti-cancer pharmaceutical composition solid oral preparation.
Advantageous effects
When the invention is used for in vitro anti-tumor test of the bamboo-cypress seed oil, the surprisingly found bamboo-cypress seed oil has the inhibition activity on tumor cells, in particular to gastric cancer cell strains, breast cancer cell strains (MCF-7), cervical cancer cell strains (Hela) and lung adenocarcinoma cell strains (A549). Particularly, the inhibition rates of the cervical cancer cell strain (Hela) and the lung cancer cell strain (A549) can respectively reach 85.33 percent and 93.21 percent.
Detailed Description
The technical solution of the present invention will be further described in detail with reference to specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.
Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.
Instruments and reagents:
bamboo cypress seed oil (prepared according to the method described in patent document CN 202010203003.0); CCK8 (shanghai beibo biotechnology limited); DMEM high-glucose medium (semer feishale (suzhou) instruments ltd); EDTA (pancreatin) (gibco); foetal bone Serum (Biological Industries); phosphate buffered saline; 96-well cell culture plates; a multifunctional microplate reader.
Example 1: in vitro antitumor Activity test
The test process of the gastric cancer cell line is taken as an example for explanation, and the test processes of other tumor cells [ lung adenocarcinoma cell line (A549), breast cancer cell line (MCF-7) and cervical cancer cell line (Hela) ] are the test processes of the gastric cancer cell line. The specific process is as follows:
1. test sample concentration preparation
100mg of bamboo-cypress seed oil is weighed, added into a 5mL plastic centrifuge tube and diluted to 1mL by DMSO.
The initial concentration of 100mg/mL was obtained. Then the initial concentration was diluted in DMSO in two-fold order to obtain 100mg/mL, 50mg/mL, 25mg/mL, 12.5mg/mL, 6.25mg/mL, 3.125mg/mL, 1.5625mg/mL, 0.78125mg/mL of 8 different concentration gradients, and stored in a refrigerator at 4 ℃ for further use.
2. Culture and inhibition activity test of gastric cancer cell strain and breast cancer cell strain
Respectively placing gastric cancer cell strain and breast cancer cell strain at 37 deg.C and saturated humidity, and containing 5% CO2After 24 hours of incubation in the incubator, the upper layer of the culture was aspirated when the cells were in the logarithmic phase, and after digestion with 0.25% trypsin-EDTA solution, digestion was stopped using high-sugar medium. And the cells were seeded in a 96-well plate such that the cell density was 5000 cells/well. The 96-well plate was placed in an incubator for 24 hours. The cell culture medium in the 96-well plate was then aspirated. The 96-well plate was supplemented with 100. mu.L of a high-sugar medium, and then 1. mu.L of test samples (5 multiple wells per concentration) were added to each well, followed by incubation at 37 ℃ under saturated humidity and 5% CO2After further incubation for 48h, 10. mu.L of CCK8 was added to each well and further incubation was carried out for 1-4h at 37 ℃. The absorbance value of each well at a wavelength of 450nm was measured on a multifunctional microplate reader. According to the inhibition rate%]X 100. The negative control was a mixed solution of V (high sugar medium)/V (dmso) 10: 1. The test result shows that: under the concentration of 25mg/mL, the sample has better inhibition effect on gastric cancer cell lines and breast cancer cell lines (MCF-7), and the inhibition rates (%) +/-SD reach 64.298 + -2.798 and 52.868 + -2.566 respectively.
3. Culture and inhibition activity test of cervical cancer cells and lung adenocarcinoma cell strains
Respectively placing cervical cancer cell strain and lung adenocarcinoma cell strain at 37 deg.C and saturated humidity, and containing 5% CO2Culturing in incubator for 24 hr while the cells are in logarithmic growthAt this time, the upper layer of the culture was aspirated and digested with 0.25% trypsin-EDTA solution, and then the digestion was stopped with a high-sugar medium. And the cells were seeded in a 96-well plate such that the cell density was 5000 cells/well. The 96-well plate was placed in an incubator for 24 hours. The cell culture medium in the 96-well plate was then aspirated. The 96-well plate was supplemented with 100. mu.L of a high-sugar medium, and then 1. mu.L of test samples (5 multiple wells per concentration) were added to each well, followed by incubation at 37 ℃ under saturated humidity and 5% CO2After further incubation for 48h, 10. mu.L of CCK8 was added to each well and further incubation was carried out for 1-4h at 37 ℃. The absorbance value of each well at a wavelength of 450nm was measured on a multifunctional microplate reader. According to the inhibition rate%]X 100. The negative control was a mixed solution of V (high sugar medium)/V (dmso) 10: 1. The test result shows that: under the concentration of 50mg/mL, the sample has better inhibition effect on gastric cancer cell strains and breast cancer cell strains (MCF-7), and the inhibition rates (%) + -SD respectively reach 85.33 + -3.035 and 93.207 + -2.237.
Example 2: compounding material of solid oral preparation and its preparation process
Figure BDA0002754481820000081
The preparation process comprises the following steps: mixing solid adjuvants uniformly, sieving with 60 mesh sieve for 3 times, adding herba Sabinae chinensis seed oil into 10% polyvidone solution, mixing uniformly by ultrasound, adding into sieved adjuvants, grinding, mixing uniformly, drying at 50-60 deg.C for 2-3 hr, grinding, mixing uniformly, tabletting, and forming to obtain 100 tablets of herba Sabinae chinensis seed oil with specification of 0.78 mg/tablet.
Other solid oral preparations can be prepared by reference to the tablet manufacturing process.
The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (7)

1. An application of a kind of bamboo-cypress seed oil in preparing anticancer medicine;
the cancer is gastric cancer, breast cancer, cervical cancer and lung adenocarcinoma;
the bamboo-cypress seed oil is prepared by the following steps:
(1) crushing the bamboo cypress seeds to obtain bamboo cypress seed powder;
(2) steaming the bamboo cypress seed powder obtained in the step (1);
(3) extruding the steamed bamboo cypress seed powder obtained in the step (2) to obtain oil;
(4) standing and filtering the extruded oil obtained in the step (3) to obtain the bamboo-cypress seed oil;
the steam evaporation in the step (2) is realized by using water vapor, and the steam evaporation equipment is any one of a water bath kettle and a steamer; the steaming time is 5-30 minutes;
the extrusion in the step (3) is realized through an extrusion device, the extrusion pressure is 0-50 MPa, and the extrusion time is 5-60 minutes;
the content of total flavonoids in the bamboo and cypress seed oil is more than or equal to 4 percent in terms of rutin;
the content of unsaturated fatty acid in the bamboo and cypress seed oil is more than or equal to 75 percent;
the content of vitamin E in the bamboo and cypress seed oil is more than or equal to 2mg/100 g.
2. The use of claim 1, wherein the raw material of bamboo cypress seeds is bamboo cypress nuts.
3. The use of claim 1, wherein the particle size of the bamboo cypress seed powder in step (1) is 20-70 mesh.
4. The use of claim 1, wherein the steaming time in step (2) is 10 to 25 minutes.
5. The use of claim 1, wherein the steaming time in step (2) is 18 to 22 minutes.
6. The use according to claim 1, wherein the fine powder of steamed bamboo and cypress seeds obtained in step (2) is artificially pressed into a fixed shape in a fixed die before extrusion.
7. The use of claim 1, wherein the standing time in the step (4) is 6-48 hours;
and (4) filtering by using any one of a Buchner funnel and a sand core funnel.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106946903A (en) * 2017-04-06 2017-07-14 澳门大学 One class drop diterpene compound, its extraction separation method and its application in antineoplastic is prepared
CN111713687A (en) * 2019-03-21 2020-09-29 厦门稀土材料研究所 Preparation method and application of bamboo-cypress seed oil

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108192721A (en) * 2017-11-22 2018-06-22 广东海洋大学 A kind of method for extracting podocarpus nagi nut oil

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106946903A (en) * 2017-04-06 2017-07-14 澳门大学 One class drop diterpene compound, its extraction separation method and its application in antineoplastic is prepared
CN111713687A (en) * 2019-03-21 2020-09-29 厦门稀土材料研究所 Preparation method and application of bamboo-cypress seed oil

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