WO2023082217A1 - Utilisation d'un extrait aqueux de cordyceps militaris avec sélénium nanométrique pour réduire une lésion de radiothérapie et d'un agent protecteur - Google Patents

Utilisation d'un extrait aqueux de cordyceps militaris avec sélénium nanométrique pour réduire une lésion de radiothérapie et d'un agent protecteur Download PDF

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WO2023082217A1
WO2023082217A1 PCT/CN2021/130490 CN2021130490W WO2023082217A1 WO 2023082217 A1 WO2023082217 A1 WO 2023082217A1 CN 2021130490 W CN2021130490 W CN 2021130490W WO 2023082217 A1 WO2023082217 A1 WO 2023082217A1
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selenium
nano
cordyceps militaris
radiotherapy
water extract
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PCT/CN2021/130490
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English (en)
Chinese (zh)
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陈填烽
刘畅
李海伟
陈义康
李绿漪
陈雯婷
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广东暨创硒源纳米研究院有限公司
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Priority to PCT/CN2021/130490 priority Critical patent/WO2023082217A1/fr
Publication of WO2023082217A1 publication Critical patent/WO2023082217A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/04—Sulfur, selenium or tellurium; Compounds thereof
    • 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
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/062—Ascomycota
    • A61K36/066—Clavicipitaceae
    • A61K36/068—Cordyceps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Definitions

  • the patent relates to the technical field of compound medicine, in particular to the application of nano-selenium Cordyceps militaris water extract in reducing radiotherapy damage and its protective agent.
  • inorganic selenium In terms of acute toxicity (LD 50 ), inorganic selenium is 15 mg/kg, organic selenium is 35 mg/kg, and nano-selenium is 113 mg/kg. Therefore, using biotransformation to convert inorganic selenium into highly active organic selenium is one of the safest and most effective means of selenium products. With the development of nanotechnology, nano-selenium has emerged. Compared with inorganic selenium and organic selenium, elemental selenium nano-selenium (SeNPs) has the characteristics of low toxicity, high bioavailability, high reaction yield and easy surface modification, and has attracted widespread attention. .
  • SeNPs elemental selenium nano-selenium
  • nano-selenium Due to the low toxicity of nano-selenium, its absorption and tolerance concentration by organisms is also significantly higher than that of inorganic selenium, so nano-selenium is safer and more efficient in the biotransformation process. Our previous studies have confirmed that nano-selenium can reverse the kidney damage caused by cisplatin and the damage of islet cells caused by fatty acids.
  • Cordyceps militaris belongs to the phylum Fungi, Ascomycetes, Sclerotinia, Spheroides, Ergotaceae, and Cordyceps.
  • Cordycepin, adenosine, and cordycepic acid are the main components of Cordyceps militaris to play its role. Studies have shown that cordycepin is an important active ingredient of Cordyceps militaris, and the content of cordycepin in Cordyceps militaris is dozens of times that of Cordyceps sinensis, and it is the main source of cordycepin.
  • Cordyceps militaris has a variety of physiological activities, including anti-virus, anti-inflammation, anti-oxidation, anti-aging, anti-tumor, anti-proliferation, anti-radiation, nerve protection, kidney protection, liver protection, and immunity enhancement.
  • Cordyceps militaris As a substitute for wild Cordyceps, Cordyceps militaris has a large market space. The artificial cultivation of Cordyceps militaris has low investment, high benefit, short production cycle and can be cultivated throughout the year. Cordycepin has been confirmed to have biological activities such as anti-inflammation, anti-tumor, anti-oxidation and inhibition of cell differentiation.
  • Cordycepin can play an anti-inflammatory role in lipopolysaccharide-induced macrophage injury in part by inhibiting the inflammatory response mediated by NLRP3 inflammasome and cyclooxygenase 2.
  • Cordycepin plays a protective role in myocardial injury by regulating NF- ⁇ B signaling pathway.
  • Cordyceps militaris has obvious effects in inhibiting the proliferation of cancer cells and inducing the apoptosis of cancer cells.
  • the selenium content in the common Cordyceps militaris fruit body is very low, and adding nano-selenium in the medium, through the biotransformation of Cordyceps militaris, the selenium content in Cordyceps militaris will be greatly increased, and basically exist in the fruit body in the form of organic selenium , which is easily absorbed and utilized by the human body.
  • the current measures for radiotherapy protection mainly include pre-radiation protective agents and post-radiation therapeutic agents, including chemical radioprotectants such as chemical compounds, plant and herbal protective agents, cytokines, and mesenchymal stem cell radiation therapy agents.
  • Amifostine and Palifermin are the only drugs approved by the U.S. Food and Drug Administration (FDA) for clinical radiotherapy protection, but these two drugs can cause hypotension, hypocalcemia, and nausea Adverse reactions such as vomiting have multiple toxic and side effects. Due to the short half-life of amifostine in the body, the protective effect can only be achieved when it is used immediately before chemotherapy, and the drug response is closely related to the time of administration. Therefore, amifostine must be administered 30 minutes before chemotherapy, the effective time window is short, and the price is expensive.
  • FDA Food and Drug Administration
  • nano-selenium Cordyceps militaris has applied for a patent before, and the patent number is ZL201811313225.7. Based on the cultivation method of this patent, the inventor found through further research that nano-selenium Cordyceps militaris has the excellent characteristics of both Cordyceps militaris and selenium , not only can be used as food, but also has value in medical applications, which needs to be proved by experiments.
  • the primary purpose of the present invention is to find the application of nano-selenium cordyceps militaris water extract in reducing radiation damage, which has good free radical scavenging and anti-oxidation effects, has good protective effect on radiation damage to the body, and has a long effective protection time and no adverse reactions. very small.
  • Another object of the present invention is to provide a protective agent of nano-selenium Cordyceps militaris water extract, which can reduce the damage of human cells in radiotherapy, and especially protect human myocardial damage caused by radiotherapy.
  • nano-selenium Cordyceps militaris has the excellent properties of Cordyceps militaris and selenium, and is a natural medicine with good pharmacological activity. More importantly, it has a good effect on antagonizing radiotherapy damage. Compared with general protective agents, the toxicity is basically It can be ignored, and it is a new type of protective agent worthy of research and development. Therefore, nano-selenium Cordyceps militaris has protective activity on myocardial injury caused by radiotherapy, which provides a new scientific basis for the development of efficient and safe radiotherapy protective agents.
  • the invention provides the application of nano-selenium Cordyceps militaris water extract in reducing radiotherapy damage.
  • the nano-selenium Cordyceps militaris water extract is a water extract obtained by crushing the nano-selenium Cordyceps militaris fruiting body, adding water and ultrasonically extracting it, and is a hyperglandular Application of aqueous extract of Cordyceps militaris with nano-selenium content in reducing radiation injury.
  • nano-selenium Cordyceps militaris water extract has free radical scavenging and anti-oxidation effects, and reduces cell damage during radiotherapy.
  • the content of adenosine in the water extract of nano-selenium Cordyceps militaris is 0.21%-0.28%
  • the content of total selenium is 80.17-198.02 ⁇ g/g
  • the proportion of selenocystine to total selenium is 17.23% ⁇ 49.07%.
  • the present invention also provides a protective agent capable of reducing radiotherapy damage, comprising the above-mentioned aqueous extract of nano-selenium Cordyceps militaris. Specifically, it is the nano-selenium Cordyceps militaris aqueous extract containing high adenosine content.
  • the content of adenosine in the water extract of nano-selenium Cordyceps militaris is 0.21%-0.28%
  • the content of total selenium is 80.17-198.02 ⁇ g/g
  • the proportion of selenocystine to total selenium is 17.23% ⁇ 49.07%.
  • the protective agent can be taken as an oral agent.
  • the present invention proves through a large number of experiments that under the same selenium content, the nano-selenium Cordyceps militaris aqueous extract has the strongest antioxidant property. Specifically, at the in vitro level, the inventors compared sodium selenite, sodium selenate, nano-selenium, selenocystine, selenomethionine, methylselenocysteine and nano-selenium Cordyceps militaris water extract, etc. Different forms of selenium have the ability to scavenge ROS caused by radiotherapy.
  • the aqueous extract of nano-selenium Cordyceps militaris has the strongest scavenging ability, and the scavenging ability is increased by 20%-90%.
  • the present inventor has also investigated the scavenging ability of the aqueous extract of Cordyceps militaris and the aqueous extract of nano-selenium Cordyceps militaris to the ROS that radiotherapy causes, the result also finds that the aqueous extract of nano-selenium Cordyceps militaris is stronger than that of aqueous extract of Cordyceps militaris.
  • the scavenging ability, the ROS scavenging ability of the nano-selenium Cordyceps militaris water extract is 1.3-2.1 times higher than that of the Cordyceps militaris water extract.
  • the present inventors found the application of nano-selenium Cordyceps militaris water extract with high adenosine content in reducing radiotherapy damage. Specifically, it is applied to human organs or muscles to reduce radiotherapy damage. It has a certain radiation protection effect, low toxicity, strong cell repair effect, and has a good protective effect on the body's radiation damage.
  • the nano-selenium cordyceps militaris water extract of the present invention has high selenium content, good free radical scavenging and anti-oxidation effects, and good protective effect on myocardial damage caused by radiotherapy. Compared with existing general drugs, the effective time window is significantly larger Growth, adverse reactions are almost negligible, and the price is lower.
  • the invention greatly expands the application field of the nano-selenium cordyceps militaris, and has broad medical drug application prospects.
  • the raw materials of the protective agent are cheap and easy to obtain, the synthesis and purification steps are highly operable, and the synthesis scale is easy to expand, so as to realize the commercialization and application of the protective agent drug.
  • Figure 1 The toxicity of different radiotherapy doses to H9C2 cells.
  • Figure 2 It is the photo of H9C2 cells and the cell survival rate under the radiation dose of 8Gy.
  • FIG. 3 Nano selenium Cordyceps militaris water extract reduces the rise of ROS caused by radiotherapy.
  • Figure 4 It is a graph showing the change of mitochondrial membrane potential in H9C2 cells.
  • Figure 5 It is a graph showing changes in body weight of mice.
  • Figure 6 Changes in the proportion of macrophages in the mouse spleen.
  • Fig. 7 It is a graph showing the change of Mn-SOD and cTn in mouse serum.
  • the invention provides the application of nano-selenium Cordyceps militaris water extract in reducing radiotherapy damage, which is an application of nano-selenium Cordyceps militaris water extract with high adenosine content in reducing radiotherapy damage. Specifically, it is applied to reducing radiotherapy damage to human organs or muscles.
  • the nano-selenium Cordyceps militaris water extract has free radical scavenging and anti-oxidation effects, and reduces cell damage during radiotherapy. For example, it can protect human heart muscle damage caused by radiotherapy.
  • the present invention also provides a protective agent capable of reducing radiotherapy damage, comprising the above-mentioned aqueous extract of nano-selenium Cordyceps militaris. Specifically, it is the nano-selenium Cordyceps militaris aqueous extract containing high adenosine content.
  • Described a kind of protective agent that can reduce radiotherapy damage is obtained by adopting the following method steps:
  • step (2) adding the polysaccharide composite nano-selenium and the Cordyceps militaris seed liquid prepared in step (1) to the fruiting body of Cordyceps militaris to continue culturing to obtain the fruiting body of the nano-selenium Cordyceps militaris;
  • the content of adenosine in the nano-selenium Cordyceps militaris water extract is 0.21%-0.28%, the content of total selenium is 80.17-198.02 ⁇ g/g, wherein, the proportion of selenocystine to total selenium is 17.23%-49.07% %.
  • the present invention provides nano-selenium Cordyceps militaris water extract in reducing radiotherapy damage. In particular, it has a protective effect on human myocardial injury caused by radiotherapy.
  • DPPH working solution (180 ⁇ L) was mixed with 20 ⁇ L of sodium selenite, sodium selenate, selenocystine, aqueous extract of Cordyceps militaris, aqueous solution of aqueous extract of nano-selenium Cordyceps militaris and positive control Trolox respectively.
  • the OD values at different times were monitored under a fluorescent microplate reader, and read continuously for 30 minutes.
  • the concentration of 0.781-25 ⁇ g/mL was selected to repeat the above experiment. It can be observed that at the drug concentration of 6.25 ⁇ g/mL, the protective effect of nano-selenium Cordyceps militaris is the best, which is higher than that of ordinary Cordyceps.
  • Nano-selenium Cordyceps militaris water extract reduces ROS caused by X-Ray.
  • H9C2 cells were inoculated in a 6 cm dish at a density of 5 ⁇ 10 4 cells/mL and incubated for 24 hours.
  • a certain amount of common Cordyceps water extract and The nano-selenium Cordyceps militaris water extract was incubated for 24 hours and given a radiation dose of 8Gy.
  • the cells were collected after 48 hours of incubation, and the fluorescence intensity was detected by flow cytometry after incubation of DCF probe for 30 minutes.
  • JC-1 detects changes in mitochondrial membrane potential.
  • H9C2 cardiomyocytes in the logarithmic growth phase were inoculated in 6 cm cell culture dishes until they adhered to the wall for 24 hours, and then added drugs to incubate the cells for 12 hours; after the cells were irradiated with X-rays (8Gy), they were placed in an incubator for 48 hours. After the cells were collected, 200 ⁇ L of 0.9% NaCl solution containing JC-1 probe (10 ⁇ g/mL) was added, and the cells were incubated at 37° C. for 30 min. Changes in mitochondrial membrane potential were detected by flow cytometry.
  • mice Normal Balb/c mice were given 0.5 mg/kg common Cordyceps militaris water extract and nano-selenium Cordyceps militaris water extract by intragastric administration every day, and a radiation dose of 5Gy was given every week for a total of 4 times of radiotherapy. 4 weeks of dosing. During this period, the body weight and other indicators of the mice were detected. After the experiment, the changes in the proportion of macrophages in the spleen of the mice were detected, and the changes of SOD and myocardial enzymes in the serum were detected.
  • the nano-selenium Cordyceps militaris water extract of the present invention has a good effect in reducing the damage of radiotherapy, specifically can reduce the damage of radiotherapy to human organs or muscles, and can significantly down-regulate the rise of ROS caused by radiotherapy, and the toxicity It has a strong cell repair effect, especially to protect human myocardial damage caused by radiotherapy.
  • the water extract of selenium-enriched Cordyceps militaris has high selenium content, has good free radical scavenging and anti-oxidation effects, and has a good protective effect on myocardial damage caused by radiotherapy. Compared with existing general drugs, the effective time window is greatly increased. Adverse reactions are almost negligible, and the price is lower.
  • the invention greatly expands the application field of the nano-selenium cordyceps militaris, and has broad medical drug application prospects.

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Abstract

Un agent protecteur contenant un extrait aqueux de Cordyceps militaris avec sélénium nanométrique, et une utilisation de l'extrait aqueux de Cordyceps militaris avec sélénium nanométrique pour réduire une lésion de radiothérapie. L'extrait aqueux de Cordyceps militaris avec sélénium nanométrique peut réduire la lésion provoquée par une radiothérapie à des organes ou des muscles humains, réduit l'augmentation des ROS causée par la radiothérapie, présente une faible toxicité, a un effet de réparation cellulaire très puissant, et joue également un rôle dans la protection contre une lésion myocardique humaine provoquée par la radiothérapie.
PCT/CN2021/130490 2021-11-13 2021-11-13 Utilisation d'un extrait aqueux de cordyceps militaris avec sélénium nanométrique pour réduire une lésion de radiothérapie et d'un agent protecteur WO2023082217A1 (fr)

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