CN117224656A - Application of functionalized nano-selenium hydrosol in anti-tumor aspect - Google Patents

Application of functionalized nano-selenium hydrosol in anti-tumor aspect Download PDF

Info

Publication number
CN117224656A
CN117224656A CN202210637693.XA CN202210637693A CN117224656A CN 117224656 A CN117224656 A CN 117224656A CN 202210637693 A CN202210637693 A CN 202210637693A CN 117224656 A CN117224656 A CN 117224656A
Authority
CN
China
Prior art keywords
selenium
polysaccharide protein
use according
cordyceps sinensis
selenite
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.)
Pending
Application number
CN202210637693.XA
Other languages
Chinese (zh)
Inventor
黄家兴
董晓莉
陆家谦
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hong Kong Polytechnic University HKPU
Original Assignee
Hong Kong Polytechnic University HKPU
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Hong Kong Polytechnic University HKPU filed Critical Hong Kong Polytechnic University HKPU
Priority to CN202210637693.XA priority Critical patent/CN117224656A/en
Priority to PCT/CN2023/092664 priority patent/WO2023236700A1/en
Publication of CN117224656A publication Critical patent/CN117224656A/en
Pending legal-status Critical Current

Links

Classifications

    • 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
    • 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
    • A61K36/07—Basidiomycota, e.g. Cryptococcus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/02—Peptides of undefined number of amino acids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • 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)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Mycology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Inorganic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The disclosure relates to preparation and anticancer application of Cordyceps sinensis mycelium Cs4 polysaccharide protein functionalized nano-selenium hydrosol.

Description

Application of functionalized nano-selenium hydrosol in anti-tumor aspect
Technical Field
The invention relates to preparation and functionalization of cordyceps sinensis mycelium Cs4 water-soluble polysaccharide protein nano-selenium (Cs 4-SeNPs) hydrosol and application thereof in anti-tumor aspect.
Background
Selenium is one of microelements necessary for health of human and animals, and has wide physiological functions (e.g. anti-tumor, antioxidant, antiaging, immunity improving, cell repair protecting, etc.). About 40 countries are in areas with selenium deficiency, and a plurality of provinces in China are in areas with selenium deficiency or low selenium, and the areas are areas with high incidence rate of tumor, liver diseases, cardiovascular diseases and the like, and selenium supplement is needed. However, as a nutritional supplement or cancer preventive, the beneficial and toxic dosage ranges of selenium are extremely narrow, which is prone to cause selenium poisoning, severely limiting the potential of selenium for disease control. Selenium toxicity depends on its chemical form. In recent years, nano selenium obtained by utilizing a nano technology has high biocompatibility, low toxicity and remarkable biological activity, and becomes a new research hot spot and a new favor for supplementing selenium.
The preparation method of nano elemental selenium generally adopts a reduction method, and utilizes the oxysalt or oxide of selenium to obtain elemental selenium through various reducing agents, and simultaneously uses a modifier or a regulator to modify and regulate the particle size and morphology, thereby obtaining an ideal product. However, the existing preparation methods of various nano elemental selenium are all to regulate or modify the morphology through various regulating agents or modifying agents, and the application needs of the nano elemental selenium are not considered. In addition, none of the regulating agents or the modifying agents is a material with biological activity, and can not effectively improve the biological activity of nano elemental selenium, especially the anti-tumor activity, namely the obtained product is not functionalized nano elemental selenium.
The nano selenium is successfully functionalized by using the cordyceps mycelium Cs4 water-soluble polysaccharide protein, namely, the cordyceps mycelium Cs4 water-soluble polysaccharide protein with the activities of resisting oxidation, resisting tumors, promoting bone growth and the like is introduced into a liquid phase nano selenium system to prepare Cs4 polysaccharide protein functionalized nano selenium (Cs 4-Sepps), so that the bioavailability and the bioactivity of selenium are improved, the cytotoxicity of selenium is reduced, and the remarkable in-vivo and in-vitro bone growth promoting activity is verified. Therefore, the Cs4-SeNPs have great potential for promoting bone development in childhood and adolescence and preventing and treating diseases such as osteoporosis, bone loss, fracture and the like in middle-aged. See the issued patent of 'a nano selenium hydrosol and a preparation method and application thereof' (patent application number: ZL 201911215358.5).
Cancer has become one of the major diseases affecting health and longevity in humans and animals such as companion animals.
According to American Veterinary Medical Association (american veterinary association) (AVMA), cancer causes almost 50% of deaths in companion animals over 10 years of age. Common cancers for companion animals include: skin cancer, testicular cancer, abdominal cancer, and bone cancer. Cancers commonly found in companion animals are also common in humans, such as lymphomas, melanomas, and osteosarcomas.
With the acceleration of urban progress in China and the rapid increase of the population numbers of empty-nest old people, empty-nest young people and the like, the population numbers for raising companion animals are rapidly growing in recent years. Along with the improvement of living standard and consumption, people pay more attention to the health of companion animals, and the companion animal health care product market is developed at a high speed, so that the potential is huge. The chance of cancer of the companion animal is comparable with that of human beings, but most of tumor treatment of the companion animal only adopts human medicines, and the fact proves that the specificity is not high and the price is high.
There is a need in the art for inexpensive nano-selenium drugs or health care products that can be conveniently used for the prevention and treatment of tumors in humans and animals, such as companion animals.
Disclosure of Invention
In one aspect, the invention provides the use of a complex comprising a Cordyceps sinensis mycelium Cs4 polysaccharide protein functionalized nano-selenium hydrosol for the preparation of a formulation for treating a tumor in a subject in need thereof.
In one embodiment, the subject is a human.
In one embodiment, the subject is a companion animal.
In one embodiment, companion animals include cats and dogs.
In one embodiment, the tumor includes osteosarcoma and malignant melanoma.
In one embodiment, the selenium is from selenium dioxide or selenite.
In one embodiment, the selenite is sodium selenite.
In one embodiment, the formulations prepared from the complexes of the invention may be injections or sprays.
In one embodiment, the preparation method of the cordyceps mycelia Cs4 polysaccharide protein functionalized nano-selenium hydrosol comprises the following steps:
(a) Extracting Cordyceps sinensis mycelia Cs4 with water to obtain Cordyceps sinensis mycelia Cs4 polysaccharide protein solution;
(b) Adding a selenium dioxide solution or selenite solution into the cordyceps sinensis mycelium Cs4 polysaccharide protein solution obtained in the step (a), and uniformly mixing to obtain a mixed solution A;
(c) Dripping a reducing vitamin C solution into the mixed solution A, and uniformly mixing to obtain a mixed solution B;
(d) And (3) after the mixed solution B is completely reacted, obtaining the cordyceps sinensis mycelium Cs4 polysaccharide protein functionalized nano-selenium hydrosol through dialysis.
In one embodiment, the molar ratio of vitamin C to selenium dioxide or selenite in mixed solution B is 5:1.
In one embodiment, in the mixed solution B, the concentration of the cordyceps sinensis mycelium Cs4 polysaccharide protein is 60-6000 mg.L -1 The concentration of vitamin C is 1.0-100 mmol.L -1 The concentration of selenium dioxide or selenite is 0.2-20mmol.L - 1。
In one embodiment, in the mixed solution B, the concentration of the cordyceps sinensis mycelium Cs4 polysaccharide protein is 300-1200 mg.L -1 The concentration of vitamin C is 10.0-100 mmol.L -1 The concentration of selenium dioxide or selenite is 2.0-20 mmol.L -1 。
In a second aspect, the present invention provides a formulation comprising a Cordyceps sinensis mycelium Cs4 polysaccharide protein functionalized nano-selenium hydrosol.
In one embodiment, the formulation is an injection or spray.
Drawings
Fig. 1: cs4-Senps, PTR-Senps and Chitosan-Senps have antiproliferative effects on OSCA-8 and CGMD-5 cells.
Fig. 2: DNA flow cytometry analysis of the effect of Cs4-SeNPs on OSCA-8 and CGMD-5 cell cycle distribution changes.
Fig. 3: the everting capacity of Cs4-SeNPs to OSCA-8 and CGMD-5 cell membrane phosphatidylserine is discussed.
THE ADVANTAGES OF THE PRESENT INVENTION
Cs4-Senps can be used in various forms such as injection and spray.
Cordyceps sinensis mycelium Cs4 polysaccharide protein is rich in hydrophilic hydroxyl (-OH) and amino (-NH) groups, so that the water solubility of nano selenium is improved, the affinity of selenium to cancer cells is enhanced, the uptake of nano selenium by tumor cells is improved, and the overall treatment effects of reducing the dosage, improving the curative effect and reducing the toxic and side effects are achieved.
Detailed Description
Cordyceps sinensis (Cordyceps sinensis) is a precious medicinal fungus and has been used as a tonic and therapeutic Chinese medicine in China for over 700 years. Since wild Cordyceps sinensis is very rare and expensive, a Cordyceps sinensis mycelium (Cs 4) successfully isolated at the 4 th time of China academy of sciences has a very great contribution to commercial production and scientific research in the past 40 years. Pharmacological and clinical studies have shown that the polysaccharide protein is one of the main bioactive components of Cs4, and has wide health promoting and therapeutic effects, such as immunomodulation, anti-tumor and bone protection. The Cordyceps sinensis mycelium Cs4 polysaccharide protein contains at least one of water-soluble polysaccharide, polysaccharide protein and protein, for example, water-soluble polysaccharide protein with total sugar content of about 41.5% and protein content of about 23.6%, wherein polysaccharide part is mainly composed of glucose, mannose and galactose, and contains small amount of mannose, arabinose and xylose.
The existing preparation methods of various nano elemental selenium are all to regulate and control or modify the morphology through various regulating and controlling agents or modifying agents to obtain products with ideal particle size and morphology, but the obtained products are not functionalized nano elemental selenium. In order to solve the problem, the invention takes the cordyceps mycelium Cs4 polysaccharide protein with wide biological activity as the functional factor of nano-selenium to obtain the cordyceps mycelium Cs4 polysaccharide protein functional nano-selenium hydrosol with anticancer effect.
In a first aspect, the present invention provides the use of a complex comprising a nano-selenium hydrosol of cordyceps mycelium Cs4 polysaccharide protein for the preparation of a formulation for the treatment of a tumor in a subject in need thereof.
In one embodiment, a subject in need thereof may include various animals, such as mammals. Formulations comprising one or more of the presently disclosed formulations may be administered to a mammal using a variety of routes. Routes of administration suitable for methods of treating cancer as disclosed herein include both local and systemic administration.
In one embodiment, the subject is a companion animal. Companion animals have well known meanings in the art, such as common cats and dogs.
In one embodiment, the formulations prepared from the complexes of the invention are useful in the treatment of common forms of cancer, including lung cancer, brain cancer, central nervous system cancer, breast cancer, colon cancer, leukemia, myeloma, prostate cancer, and ovarian cancer. Sarcomas are malignant tumors derived from connective tissue or mesenchymal cells. Blastomas are common malignant tumors that resemble immature or embryonic tissue. Lymphomas and leukemias are malignant tumors derived from hematopoietic (blood-forming) cells. In a preferred embodiment, the pharmaceutical compositions of the invention are formulated for use in the treatment of osteosarcoma and malignant melanoma.
In one embodiment, the selenium is from selenium dioxide or selenite. Selenite includes sodium selenite, potassium selenite, barium selenite, zinc selenite, etc.
In one embodiment, the formulations prepared from the complexes of the invention may be produced using any of a variety of processes including, but not limited to, conventional mixing, dissolving, granulating, coating, emulsifying, encapsulating, and lyophilizing. The formulations may take any of a variety of forms including, but not limited to, hydrosols, sterile solutions, suspensions, emulsions, lyophilized formulations, tablets, drop pills, pellets, capsules, powders, syrups, elixirs or any other dosage form suitable for administration. The preferred dosage form is an injection or spray.
Formulations prepared from the complexes disclosed herein may optionally include, but are not limited to, other pharmaceutically acceptable components (or pharmaceutical components), including, but not limited to buffers, preservatives, tonicity adjusting agents, salts, antioxidants, tonicity adjusting agents, physiological substances, pharmacological substances, fillers, emulsifying agents, wetting agents, sweetening or flavoring agents and the like. Various buffers and methods of adjusting pH can be used to prepare the formulations disclosed herein. Such buffers include, but are not limited to, acetate buffers, borate buffers, citrate buffers, phosphate buffers, neutral buffered saline, and phosphate buffered saline.
In a second aspect, the present invention provides a formulation comprising a Cordyceps sinensis mycelium Cs4 polysaccharide protein functionalized nano-selenium hydrosol.
In one embodiment, the preparation method of the cordyceps mycelia Cs4 polysaccharide protein functionalized nano-selenium hydrosol comprises the following steps:
(a) Extracting Cordyceps sinensis mycelia Cs4 with water to obtain Cordyceps sinensis mycelia Cs4 polysaccharide protein solution;
(b) Adding a selenium dioxide solution or selenite solution into the cordyceps sinensis mycelium Cs4 polysaccharide protein solution obtained in the step (a), and uniformly mixing to obtain a mixed solution A;
(c) Dripping a reducing vitamin C solution into the mixed solution A, and uniformly mixing to obtain a mixed solution B;
(d) And (3) after the mixed solution B is completely reacted, obtaining the cordyceps sinensis mycelium Cs4 polysaccharide protein functionalized nano-selenium hydrosol through dialysis.
In the step (a), the cordyceps sinensis mycelia Cs4 polysaccharide protein can be prepared by the following method: adding water into the cordyceps sinensis mycelia Cs4 according to the weight ratio of the cordyceps sinensis mycelia Cs4 to the water of 1:20, heating to 95-100 ℃, extracting for 2 hours, and then taking filtrate; repeatedly extracting the filter residue for 2 hours according to the previous steps, combining the filtrates obtained by the two extractions, dialyzing for 24 hours, and obtaining the Cordyceps sinensis mycelium Cs4 polysaccharide protein with the molecular weight cut-off of 8,000.
In the step (C), the molar ratio of the vitamin C to the selenium dioxide or selenite in the mixed solution B is 5:1.
The cordyceps sinensis mycelium Cs4 polysaccharide protein has specificity in chemical structure, and the polysaccharide part has polyhydroxy structure, so that the polysaccharide part has strong physical adsorption effect on nano selenium, further aggregation and precipitation of nano selenium are avoided, the surface of the nano selenium is effectively modified, and good particle size regulation and stabilization effects are exerted. In addition, the cordyceps sinensis mycelium Cs4 polysaccharide protein is rich in hydrophilic hydroxyl (-OH) and amino (-NH) groups, so that the water solubility of nano-selenium is improved, the affinity between nano-selenium and tumor cells is enhanced, the uptake of nano-selenium by the tumor cells is greatly improved, and the overall treatment effects of reducing the dosage, improving the curative effect and reducing the toxic and side effects are achieved. In addition, the novel nano-selenium prepared by the method can be stored in a liquid phase in a hydrosol form for a long time, has good dispersibility and stability, and can be used for various dosage forms such as injection, spraying and the like.
In one embodiment, the cordyceps mycelia Cs4 polysaccharide protein solution, the selenium dioxide solution or the selenite solution and the vitamin C solution adopted by the invention are all prepared by taking water as a solvent. In the step (d), whether the mixed solution B is completely reacted or not can be judged according to the color of the product, for example, the reaction is complete when the red color in the product is no longer deepened. In addition, when the vitamin C solution is added to the mixed solution a, the vitamin C solution may be added dropwise to the mixed solution a. In the step (C), the vitamin C is reduced vitamin C.
As the preparation method of the nano selenium hydrosol, in the mixed solution B, the concentration of the polysaccharide protein of the Cordyceps sinensis mycelium Cs4 is 60-6000 mg.L -1 The concentration of selenium dioxide or selenite is 0.2-20mmol.L -1 The concentration of vitamin C is 1.0-100 mmol.L -1 . The concentration volume required by the raw materials can be calculated by a technician according to the novel nano selenium hydrosol with different concentration requirements.
In one embodiment, the molar ratio of vitamin C to selenium dioxide or selenite in mixed solution B is 5:1.
In a preferred embodiment, the concentration of the polysaccharide protein of the Cordyceps sinensis mycelia Cs4 in the mixed solution B is 300-1200mg.L -1 The concentration of vitamin C is 10.0-100 mmol.L -1 The concentration of selenium dioxide or selenite is 2.0-20 mmol.L -1 。
The novel nano selenium hydrosol can be stored in a sol form at the temperature of 2-10 ℃ and has the storage period of 30-60 days.
Examples
The materials and reagents used in this example are commercially available and the methods used are well known in the art.
Example 1
The present example prepared Cs4-SeNPs, pleurotus Tuber Regium polysaccharide protein functionalized nano-selenium (PTR-SeNPs) and Chitosan functionalized nano-selenium (Chitosan-SeNPs) hydrosol. The specific method comprises the following steps: at normal temperature and normal pressure (15-35 ℃,1 standard atmospheric pressure), the mass concentration is 2.5 g.L -1 6.0mL of Cordyceps sinensis mycelium Cs4 polysaccharide protein solution, 6.0mL of Pleurotus Tuber Regium polysaccharide protein solution, and 3.0mL of chitosan solution are respectively added into 25mL volumetric flask filled with 10mL of double distilled water, and then added into the volumetric flask with concentration of 0.025 mol.L -1 2.0mL of sodium selenite solution, gently shaking to mix thoroughly, and then adding dropwise the solution with the concentration of 0.1 mol.L -1 2.5mL of vitamin C solution is added dropwise while being gently and gently shaken, water is added to a volume of 25mL after the addition is completed, and 3 functionalized nano selenium hydrosol is obtained after the red color is not deepened. The selenium content (concentration about 1.0-1.5 mmol/L) of the product was measured by the nitrifying ICP method after 24 hours dialysis (molecular weight cut-off 8,000).
Example 2
The half-maximal Inhibitory Concentration (IC) of cells commonly used in the art was used 50 ) Are used to evaluate the ability of a drug to inhibit tumor proliferation. The antiproliferative effect of various nano-selenium species on canine osteosarcoma (OSCA-8) cancer cells and canine oral malignant melanoma (CMGD-5) was assessed by MTS cell proliferation assay (BMG Labtech, clariostar). MTS is an MTT (thiazole blue) analogue, and in the presence of a coupling agent, dehydrogenases in living cells are converted into a liquid soluble formazan compound, and the absorption value of the formazan compound at 490nm can be directly measured on a 96-well plate, and the color depth and the cell number are in a linear relationship, so that the proliferation condition of cells is reflected. As shown in FIGS. 1A-F, 3 functional nano-selenium has different degrees of inhibition on proliferation of OSCA-8 and CMGD-5 cells, and Cs 4-Sepps 4 can exert remarkably strongest antiproliferative effect (OSCA-8: IC), which is distinct from single Cordyceps mycelium Cs4 polysaccharide protein, tiger milk mushroom polysaccharide protein and chitosan 50 ＝3.82μM；CMGD-5：IC 50 ＝3.28μM)。
Example 3
The ability of Cs4-SeNPs (3-40. Mu.M) to induce OSCA-8 and CMGD-5 cell cycle arrest and apoptosis was further analyzed by DNA flow cytometry. As shown in FIGS. 2A-H, the percentage of cells that were the cells that caused the apoptosis peak (Sub-G1) by Cs4-SeNPs increased in a dose-dependent manner, (OSCA-8: from 7.0% to 90.7% in the control group, and CMGD-5: from 5.3% to 53.0% in the control group), indicating that Cs4-SeNPs were effective in inducing apoptosis in OSCA-8 and CMGD-5 cells.
Example 4
The cell membrane changes during apoptosis, and Phosphatidylserine (PS) of normal cells is located inside the cell membrane, and PS of the cell membrane is turned from inside to outside in early stages of apoptosis. The ability of Cs4-Senps (3-40. Mu.M) to induce apoptosis in OSCA-8 and CMGD-5 cells was studied by an Annexin-V kit, as shown in FIGS. 3A-H. The Annexin-V fluorescent probe specifically binds to PS and generates green fluorescence, and the green fluorescence intensity is proportional to the degree of everting PS. The results showed that the percentage of PS eversion in canine cancer cells caused by Cs4-SeNPs was increased in a dose-effect (OSCA-8: from 4.80% to 25.6% in the control group; CMGD-5: from 7.40% to 22.1% in the control group). The result shows that the Cs4-SeNPs can effectively induce the early apoptosis of OSCA-8 and CMGD-5 cells, and the effect is concentration dependent.
In conclusion, the inventors found in experiments that Cs4-Senps was effective in inhibiting canine osteosarcoma (OSCA-8, IC 50 =3.82 μm) and canine oral malignant melanoma (CMGD-5; IC (integrated circuit) 50 =3.28 μm). In addition, we found that Cs4-SeNPs exert significantly stronger antiproliferative effects on OSCA-8 and CMGD-5 cells than other different species of nanoselenium. Further mechanism studies indicate that Cs4-SeNPs achieve antitumor activity (including sub-G1 cell population increase and phosphatidylserine eversion) mainly by inducing apoptosis of OSCA-8 and CMGD-5 cells.
The above embodiments are only for illustrating the technical solution of the present invention and not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted without departing from the spirit and scope of the technical solution of the present invention.

Claims (14)

1. Use of a complex comprising a Cordyceps sinensis mycelium Cs4 polysaccharide protein functionalized nano-selenium hydrosol for the preparation of a formulation for treating a tumor in a subject in need thereof.
2. The use according to claim 1, wherein the subject is a human.
3. The use according to claim 1, wherein the subject is a companion animal.
4. The use according to claim 3, wherein the companion animal comprises cats and dogs.
5. The use according to any one of claims 1-4, wherein the neoplasm comprises osteosarcoma and malignant melanoma.
6. The use according to any one of claims 1-4, wherein selenium is derived from selenium dioxide or selenite.
7. The use according to claim 6, wherein the selenite is sodium selenite.
8. The use according to any one of claims 1 to 4, wherein the cordyceps mycelia Cs4 polysaccharide protein is obtained by water extraction of cordyceps mycelia Cs 4.
9. The use according to any one of claims 1-4, wherein the preparation method of the cordyceps mycelia Cs4 polysaccharide protein functionalized nano-selenium hydrosol comprises the following steps:
(a) Extracting Cordyceps sinensis mycelia Cs4 with water to obtain Cordyceps sinensis mycelia Cs4 polysaccharide protein solution;
(b) Adding a selenium dioxide solution or selenite solution into the cordyceps sinensis mycelium Cs4 polysaccharide protein solution obtained in the step (a), and uniformly mixing to obtain a mixed solution A;
(c) Dripping the solution of the reducing vitamin C into the mixed solution A, and uniformly mixing to obtain a mixed solution B;
(d) And (3) after the mixed solution B is completely reacted, obtaining the cordyceps sinensis mycelium Cs4 polysaccharide protein functionalized nano-selenium hydrosol through dialysis.
10. The use according to claim 9, wherein the concentration of the polysaccharide protein of the Cordyceps sinensis mycelia Cs4 in the mixed solution B is 60-6000 mg.L -1 The concentration of the reducing vitamin C is 1.0-100 mmol.L -1 The concentration of selenium dioxide or selenite is 0.2-20mmol.L -1 。
11. The use according to claim 9, wherein the concentration of the polysaccharide protein of the Cordyceps sinensis mycelia Cs4 in the mixed solution B is 300-1200mg.L -1 The concentration of vitamin C is 10.0-100 mmol.L -1 The concentration of selenium dioxide or selenite is 2.0-20 mmol.L -1 。
12. Use according to claim 9, in the mixed solution B, the molar ratio of the reducing vitamin C to selenium dioxide or selenite is 5:1.
13. A preparation for treating tumor comprises Cordyceps sinensis mycelia Cs4 polysaccharide protein functionalized nano selenium hydrosol.
14. The formulation according to claim 13, wherein the formulation is an injection or a spray.
CN202210637693.XA 2022-06-07 2022-06-07 Application of functionalized nano-selenium hydrosol in anti-tumor aspect Pending CN117224656A (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN202210637693.XA CN117224656A (en) 2022-06-07 2022-06-07 Application of functionalized nano-selenium hydrosol in anti-tumor aspect
PCT/CN2023/092664 WO2023236700A1 (en) 2022-06-07 2023-05-08 Use of functionalized nano-selenium hydrosol in anti-tumor aspect

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202210637693.XA CN117224656A (en) 2022-06-07 2022-06-07 Application of functionalized nano-selenium hydrosol in anti-tumor aspect

Publications (1)

Publication Number Publication Date
CN117224656A true CN117224656A (en) 2023-12-15

Family

ID=89095373

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202210637693.XA Pending CN117224656A (en) 2022-06-07 2022-06-07 Application of functionalized nano-selenium hydrosol in anti-tumor aspect

Country Status (2)

Country Link
CN (1) CN117224656A (en)
WO (1) WO2023236700A1 (en)

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102895258A (en) * 2011-07-25 2013-01-30 香港理工大学 Pleurotus tuber-regium polysaccharide functionalized nanometer selenium hydrosol having anti-tumor activity and preparation method thereof
CN102327620A (en) * 2011-07-29 2012-01-25 暨南大学 Application of nano-selenium in antineoplastic drug carrier
CN107412280B (en) * 2016-02-16 2021-05-18 香港理工大学深圳研究院 Nano-selenium hydrosol with anti-tumor activity, preparation and preservation method and application
CN106860484B (en) * 2016-12-27 2021-03-26 东莞市东阳光冬虫夏草研发有限公司 Fresh cordyceps sinensis extract with anti-tumor activity and preparation method and application thereof
KR20200048880A (en) * 2018-10-31 2020-05-08 가톨릭관동대학교산학협력단 Pharmaceutical composition for prevention or treatment of melanoma comprising extracts of cordyceps militaris or fraction thereof as an active ingredient
CN110876754B (en) * 2019-12-02 2022-04-15 香港理工大学深圳研究院 Nano-selenium hydrosol and preparation method and application thereof

Also Published As

Publication number Publication date
WO2023236700A1 (en) 2023-12-14

Similar Documents

Publication Publication Date Title
KR101514470B1 (en) - ligand modified poly oxo-hydroxy metal ion materials their uses and processes for their preparation
KR20050109269A (en) A ginseng preparation using vinegar and process for thereof
US20230132001A1 (en) Immunity-boosting agent, immuno-therapeutic anti-cancer agent, and anti-cancer therapy adverse effect mitigating agent containing anthocyanin-fucoidan complex as active ingredient
CN110339350A (en) A kind of antitumor drug combination compositions and its application
CN108404137B (en) Composite nano-drug constructed by coupling nano-particle with drug component, preparation method and application
KR20140097492A (en) Ligand modified poly oxo-hydroxy metal ion materials, their uses and processes for their preparation
EP1549329B1 (en) Extract with anti-tumor and anti-poisonous activity
US20080124318A1 (en) Algae supplement and treatment method
CN117224656A (en) Application of functionalized nano-selenium hydrosol in anti-tumor aspect
JP4896332B2 (en) Bioactive composition based on the active ingredients of basidiomycetes and araceae
AU2002315075B2 (en) Black soybean polysaccharides
CN114306340B (en) Preparation method and application of cholic acid-quaternized chitosan oligosaccharide-ES 2 peptide/camptothecin conjugate
US11065293B2 (en) Pharmaceutical composition for decreasing the side effects of cancer drug, and manufacturing method and uses thereof
CN113384698B (en) Self-assembled nano-medicament for synergetic chemotherapy/acousto-photodynamic therapy and application thereof
Upase et al. A review on Phytosome loaded with novel herbal drug and their formulation, standardization and applications
JP2004010605A (en) Antitumor agent given by using mixture of deacetylated and deoxygenated mushrooms, fermented ganoderma spawn, and other mixture, and health food product, and feed additive containing the agent
KR102085582B1 (en) Composition for antitumor or inducing antitumor immune response comprising Erysimum sp. extract as effective component
CN111763185A (en) Prostaglandin derivative with epoxidized side chain, composition and use thereof
CN101537013A (en) Application of nanometer polysaccharide for preparing antineoplastic medicament
JP4245291B2 (en) Bioactive composition and method for producing the same
KR20050097710A (en) Modified ginsenoside mixture which has selective anti-lung cancer activity
CN115919884B (en) Oleuropein-polysaccharide complex capable of masking oleuropein bitter taste
CN110366419A (en) Using ginsenoside compound K for inhibiting the composition of cytotoxic anticancer agent side effect
JP7333969B2 (en) Use of Lead Borate Nanoparticles Targeting Mutant p53 Gene in Cancer Therapy and Methods of Making These Nanoparticles
KR101257909B1 (en) Pharmaceurtical compositions for the prevention or treatment of cancer containing the mixed extract of Phellinus linteus mycelium and cultured Panax Ginseng Cameyer as an active ingredient

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
REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 40096471

Country of ref document: HK