WO2018033033A1

WO2018033033A1 - Use of fullerene structure in preparation of medicament for treating anemia and complication thereof

Info

Publication number: WO2018033033A1
Application number: PCT/CN2017/097305
Authority: WO
Inventors: 王春儒; 甄明明; 贾旺; 白春礼
Original assignee: 北京福纳康生物技术有限公司; 中国科学院化学研究所
Priority date: 2016-08-15
Filing date: 2017-08-14
Publication date: 2018-02-22
Also published as: CN107753510A

Abstract

Use of a fullerene structure in preparation of a medicament for treating anemia and complications thereof. The fullerene structure comprises at least one of: fullerene, a water-soluble modified form of fullerene, a fullerene composition prepared by dispersing fullerene in an oil solution, and a derivative of fullerene.

Description

Application of fullerene structure in the preparation of drugs for treating anemia and its complications

cross reference

The invention requires the application of the full application number CN201610670474.6 submitted by the Institute of Chemistry of the Chinese Academy of Sciences to the Chinese Patent Office on August 15, 2016, the invention name is "the use of fullerenes in the preparation of medicines and the prevention or treatment of iron deficiency anemia. The priority of the Chinese Patent Application, the entire disclosure of which is hereby incorporated by reference.

Technical field

The invention belongs to the field of biomedicine, relates to the application of fullerene structure in preparing medicine for treating anemia and its complications, and particularly relates to the preparation of a fullerene structure for treating nutritional anemia, hemolytic anemia, hemorrhagic anemia and the above three The application of complications in drugs.

Background technique

According to the World Health Organization, there are about 3 billion people with different degrees of anemia in the world, and the number of people who die from various diseases caused by anemia every year is tens of millions. Among them, nutritional anemia and hemolytic anemia are the most anemia diseases with the highest degree of malignancy.

Nutritional anemia refers to a disease in which the body's hematopoietic nutrients such as iron, folic acid, vitamin B12, etc. are relatively or absolutely reduced, resulting in the formation of hemoglobin or insufficient red blood cell production, resulting in low hematopoietic function, including lack of Iron anemia and megaloblastic anemia.

(1) Iron deficiency anemia refers to the lack of stored iron that can be used to make hemoglobin in the body, and the anemia that occurs when erythropoiesis is impaired. At present, the treatment methods for iron deficiency anemia mainly include traditional Chinese medicine therapy, iron supplementation, and red blood cell transfusion. Although Chinese medicine therapy has certain effects, it involves many formulas, the active ingredients are difficult to determine, and the cycle is long; iron supplements, such as ferrous sulfate tablets, iron fumarate, etc. Although it has the effect of improving anemia, it generally cannot solve the problem of absorption of the gastrointestinal tract. The patients often have adverse symptoms such as diarrhea and vomiting; blood transfusion can quickly reduce or correct anemia, but blood transfusion is more likely to cause severe transfusion reaction and increase hepatitis. , syphilis, HIV infection opportunities, and long-term multiple blood transfusions can also cause excessive iron overload, secondary hemochromatosis.

(2) megaloblastic anemia, which is caused by abnormalities in nuclear DNA synthesis caused by folic acid or vitamin B12 deficiency or drug effects. Treatment of megaloblastic anemia requires treatment of underlying diseases, removal of causes, correction of partial eclipse and poor cooking habits, supplementation with folic acid or vitamin B12.

Another type of anemia, hemolytic anemia is the acceleration of red blood cell destruction, and the anemia that occurs when the bone marrow hematopoietic function is insufficiently compensated, generally due to erythrocyte intrinsic defects or extracellular factors, among which the former is mainly globin chain-forming anemia. The latter is mainly immune hemolytic anemia caused by immune, physical, chemical, biological and other factors.

(1) globin chain-forming anemia (also known as thalassemia or marine anemia) is a group of anemia or pathology caused by the lack or deficiency of synthesis of one or more globin chains in hemoglobin due to genetic defects. status. At present, blood transfusion is the most important measure for treating thalassemia, and the blood transfusion is used to treat thalassemia.

(2) Immunological hemolytic anemia, as the most common type of acquired hemolysis, the current treatment of this type of anemia is mainly to clear the cause, remove the cause, blood transfusion, splenectomy, immunosuppressive agents, integrated Chinese and Western medicine treatment, etc. Various deficiencies mentioned.

Another type of anemia, hemorrhagic anemia, is also common. Hemorrhagic anemia is caused by a decrease in blood volume and subsequent blood thinning, which reduces blood oxygen carrying capacity. Blood transfusion is the only reliable and rapid method to restore blood volume, but The treatment methods also have the various deficiencies mentioned above.

And anemia may affect various organs in the human body, such as the liver, kidneys and spleen. The liver is an important organ in the human body. It shoulders functions such as metabolism (including glucose metabolism, protein metabolism, fat metabolism, vitamin metabolism, hormone metabolism, etc.), bile production and excretion, detoxification, immune function, and blood coagulation. The kidney is an important organ in the human body. Its main functions are: excretion of metabolites in the body and harmful substances entering the body, maintaining water balance, maintaining electrolyte and acid-base balance in the body, regulating blood pressure, promoting erythropoiesis, and promoting vitamin D activation. The spleen is important for the human body The immune organ has multiple functions: the spleen is a blood bank, which can store blood and make blood. The spleen is like a "lymph node" of blood circulation, which can function as a blood filter. The spleen is filled with a large amount of lymphoid tissue, which can produce antibodies and opsonins. Can regulate endocrine; regulate bone marrow activity; anti-tumor.

Therefore, it is important to find a product and method for treating nutritional anemia, hemolytic anemia, blood loss anemia and three complications.

Fullerenes are another allotrope of carbon other than graphite, diamond and amorphous carbon. This type of substance refers to a cage structure composed of carbon atoms. The most abundant molecules are C ₆₀ , then C ₇₀ and C ₈₄ , followed by C ₇₆ , C ₇₈ , C _{82 ,} etc. with relatively small contents. In addition, since the inside of the carbon cage of fullerenes is a cavity structure, the internal cavity can embed different atoms, ions or clusters of atoms, which is called embedded fullerene, such as La@C ₆₀ , indicating La embedded. In the cage structure of C ₆₀ , @ denotes at, the image expresses the meaning of embedded.

The information disclosed in this Background section is only intended to provide an understanding of the general background of the invention, and should not be construed as an admission

Summary of the invention

One of the objects of the present invention is to provide a fullerene structure for the preparation of a medicament for treating nutritional anemia, hemolytic anemia, hemorrhagic anemia, and the above three complications, health foods or health care products. Another object of the present invention is to provide a medicament and method for using the above fullerene structure and treating nutritional anemia, hemolytic anemia, hemorrhagic anemia, and the above three complications. A third object of the present invention is to provide a health food or health care product which uses the above fullerene structure and improves nutritional anemia, hemolytic anemia, hemorrhagic anemia, and the above three complications.

In order to achieve the object, the present invention provides the following technical solutions:

In one aspect of the invention, the invention provides the use of a fullerene structure comprising a fullerene, a water soluble modification of the fullerene, the Fullerene structure, the Fullerene structure At least one of a fullerene composition prepared by dispersing an olefin in an oil solution and a derivative of the fullerene The drug is used for treating nutritional anemia, hemolytic anemia, hemorrhagic anemia, and complications of the above three.

In another aspect of the present invention, the present invention provides a method for treating nutritional anemia, hemolytic anemia, hemorrhagic anemia, and the above three complications, comprising the steps of: treating atrophic anemia, hemolytic anemia, An effective dose of a fullerene structure is administered to a subject of hemorrhagic anemia or the above three complications; the fullerene structure comprises fullerenes, a water-soluble modification of the fullerene, the fullerene At least one of a fullerene composition prepared by dispersing in an oil solution and a derivative of the fullerene.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for treating nutritional anemia, hemolytic anemia, hemorrhagic anemia, and the above three complications, comprising a fullerene structure as an active ingredient, and further comprising At least one of a pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent, and a pharmaceutically acceptable excipient; the fullerene structure comprising a fullerene, a water soluble modification of the fullerene, the The fullerene is dispersed in at least one of a fullerene composition prepared in an oil solution and a derivative of the fullerene.

In another aspect of the present invention, the present invention provides a health food or health care product for improving nutritional anemia, hemolytic anemia, hemorrhagic anemia, and the above three complications, comprising a fullerene structure as an active ingredient, Also included is at least one of a carrier, a diluent, and an excipient that are acceptable in a health food or health care product; the fullerene structure includes a fullerene, a water-soluble modification of the fullerene, the rich At least one of a fullerene composition prepared by dispersing an oil in an oil solution and a derivative of the fullerene.

In another embodiment, the fullerene comprises at least one of a hollow fullerene and an inlaid metal fullerene, wherein:

The hollow fullerene is one or more cage structures composed of carbon atoms of the formula C _2m , 20 ≤ m ≤ ₆₀ , for example; C ₆₀ , C ₇₀ , C _{84 ,} etc.;

The inlaid metal fullerene includes M@C _2n , M ₂ @C _2n , MA@C _2n , M ₃ N@C _2n , M ₂ C ₂ @C _2n , M ₂ S@C _2n , M ₂ O One or more of @C _2n and M _x A _3-x N@C _2n , wherein: M and A each represent a metal element and M and A are each selected from any one of Sc, Y and a lanthanide metal element. Species, 20≤n≤60; 0≤x≤3. For example: Gd@C ₈₂ . N represents a nitrogen element, C represents a carbon element, S represents a sulfur element, and lanthanide metal elements include La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu.

In another embodiment of the above application, method, pharmaceutical composition, health food or health care product, the water soluble modification of the fullerene comprises one or more fullerenes selected from the group consisting of: (1) a fullerene having a hydrophilic group modified on the surface; (2) fullerenes encapsulated by hydrophilic small biomolecules; (3) fullerenes supported by a biocompatible carrier material; (4) The formed water-soluble supramolecular system fullerene is assembled.

In another embodiment of the above application, method, pharmaceutical composition, health food or health care product, the hydrophilic group comprises one or more of a hydroxyl group, a carboxyl group, a thiol group, an amino group, and a water-soluble amino acid residue.

In another embodiment, the water-soluble amino acid residue refers to a water-soluble amino acid remaining in a part of the amino acid molecule when the fullerene is modified. The complete amino acid, ie, the amino acid residue is part of the amino acid molecule, which is an incomplete amino acid. Any part of the missing amino acid molecule is considered to be an amino acid residue, such as: loss of hydrogen on the amino group in the amino acid, loss of hydrogen or hydroxyl groups on the carboxyl group in the amino acid, and the like. Optionally, the water-soluble amino acid residue is at least one of an alanine residue, a glycine residue, a serine residue, an arginine residue, a lysine residue, and a tyrosine residue.

In another embodiment, the water-soluble modification of the fullerene is a water-soluble hydroxylated ruthenium metal fullerene, a water-soluble hydroxylated C ₆₀ , or the like. Water-soluble hydroxylated C ₇₀ , water-soluble aminated ruthenium metal fullerene, such as Gd@C ₈₂ (NH ₂ ) _n , water-soluble aminated C ₆₀ , water-soluble aminated C ₇₀ , water-soluble carboxylated ruthenium metal rich The olefin is, for example, Gd@C ₈₂ (COOH) _n , a water-soluble carboxylated C ₆₀ , a water-soluble carboxylated C ₇₀ or the like.

In another embodiment of the above application, method, pharmaceutical composition, health food or health care product, the water-soluble modification of the fullerene is of the formula C _2a (OH) _b or M@C _2d (OH) _e Where: 20 ≤ a ≤ 60, optional a is 30 or 35; 0 < b < 50, optional 0 < b < 30, and optionally b = 13, 20, 24, etc.; M is selected from the group consisting of rare earths Metal, optional rare earth metals are Gd, La, etc.; 20 ≤ d ≤ 60, optional d is 41 or 30 or 35; 0 < e < 50, optional 0 < e < 30, and optionally e = 13, 20, 24, etc.

In another embodiment of the above application, method, pharmaceutical composition, health food or health care product, the hydrophilic biological small molecule comprises at least one of an amino acid and a peptide chain.

In another embodiment, the biocompatible carrier material comprises at least at least one of a liposome, a protein, a polymer micelle, and a cell membrane carrier. One of: wherein the protein comprises albumin or transferrin; and the polymer micelle comprises at least one of polyethylene glycol lactide, polylysine, and chitosan.

In another embodiment, the above-described application, method, pharmaceutical composition, health food or health care product, the fullerene structure has an average particle diameter of from 1 to 1000 nm, and optionally has an average particle diameter of from 1 to 200 nm. The fullerene active material having a particle diameter within the above range has good biocompatibility, is easily dispersed uniformly in common excipients, is simple to prepare, has certain rigidity, is not easily deformed, and can be well circulated into hematopoietic tissue and liver tissue through blood circulation. In kidney tissue and spleen tissue.

The above application, method, pharmaceutical composition, health food or health care product. In another embodiment, the method for modifying the water-soluble modification of fullerene, the method of encapsulation, the method of loading, and the method of self-assembly are not particularly limited. The skilled person can select an appropriate method to synthesize a water-soluble modification of fullerene according to the specific composition of the fullerene and the specific requirements of the drug. E.g:

(1) Method of Physical Coating The at least one of the raw material fullerenes may be mixed with at least one of polyethylene glycol, polyvinylpyrrolidone and cyclodextrin and subjected to ball milling or ultrasonication to obtain a corresponding material. Water-soluble modifications of coated fullerenes, such as polyethylene glycol-coated fullerenes and/or polyethylene glycol-coated inlaid metal fullerenes, polyvinylpyrrolidone-coated fullerenes And/or polyvinylpyrrolidone coated inlaid metal fullerenes.

(2) Preparation of water-soluble hydroxyfullerene Liquid-liquid reaction involving phase transfer catalyst (XingG et al, J. Phys. Chem. B., 2004 (108): 11473-11479) or fullerene body Directly reacted with H ₂ O ₂ under basic conditions. The preparation of the water-soluble amino fullerene can be obtained by reacting a fullerene solid powder with H ₂ O ₂ and ammonia. Thereby, the hydrophobic and hydrophobic properties of the fullerene surface can be improved to achieve the purpose of water-soluble modification. Optionally, the method for preparing the water-soluble hydroxyfullerene comprises: (a) using an aqueous hydrogen peroxide solution (optionally, the aqueous hydrogen peroxide solution is 1-30% by mass) and sodium hydroxide solution/hydrogen peroxide Mix the potassium solution (optional, sodium hydroxide solution / potassium hydroxide solution in a mass percentage of 10-80%) (optional, according to the volume of aqueous hydrogen peroxide solution and sodium hydroxide solution / potassium hydroxide solution) The ratio is 1-10:1 mixed), fullerene is added to the mixture (optionally, 20-500 mg of fullerene is added per 10-200 ml of the mixture), and the reaction is carried out at a temperature of 50-80 ° C. Alternatively, the reaction is a stirred reaction for 4-24 h; optionally, stirring at a rate of 1000 r/min), filtered, and the filtrate is retained. (b) adding the filtrate to an excess of ethanol (optionally, the ethanol concentration is 85% - 100%), after centrifugation (optional, centrifugation speed 10000 r / min, centrifugation time 1-10 min) The precipitate was collected and the precipitate was dissolved in water to give a solution. (c) subjecting the solution obtained in the step (b) to dialysis treatment, optionally, dialysis to a conductivity of the solution at room temperature of less than 1 μs/cm; optionally, lyophilizing the solution after the dialysis In order to obtain a hydroxylated fullerene solid.

(3) Water-soluble carrier-loaded fullerene The non-covalent bond modification method can be used to add fullerenes and water-soluble carriers, such as liposomes, polymer micelles, proteins, etc., to an oily solvent through hydrophobic-hydrophobic In interaction, the fullerene active is combined with the water soluble carrier described above.

In another embodiment, the oil solution may be a single component oil or a mixed oil formed of different oil solutions. Usually vegetable oils, such as olive oil, linseed oil, sunflower oil, corn germ oil, soybean oil, sesame oil, rice bran oil, flax oil, safflower oil, walnut oil, etc., also include animal fats such as squalane.

In another embodiment, the method, the method, the pharmaceutical composition, the health food or the health care product, wherein the fullerene is dispersed in the oil solution, after the mixture of the fullerene and the oil solution is subjected to ball milling or ultrasonication The precipitate is removed by centrifugation in sequence, and then the resulting supernatant liquid is filtered to remove particles, which is obtained.

In another embodiment, the above-described application, method, pharmaceutical composition, health food or health care product, the concentration of fullerene in the fullerene composition is 0.01-100 mg/mL, and the disclosure of the range should be regarded as being within the range. All values are disclosed, optionally 0.01-0.8 mg/mL, 0.01-1 mg/mL, 0.01-10 mg/mL, 10-20 mg/mL, 20-30 mg/mL, 30-40 mg/mL, and the like.

In another embodiment, the above-mentioned application, method, pharmaceutical composition, health food or health care product, in the process of mixing fullerenes with an oil solution, mixing 0.05-1000 mg of fullerene per 1 ml of the oil solution, the range of Disclosure should be considered as disclosure of all values in the range, optionally 0.05-1 mg, 0.05-10 mg, 0.05-100 mg, and the like.

In another embodiment, the above-described application, method, pharmaceutical composition, health food or health care product is subjected to ball milling or ultrasonication for 30 min to 15 h.

In another embodiment, the above-mentioned application, method, pharmaceutical composition, health food or health care product, after the mixture is subjected to ball milling or ultrasonication, before centrifugation, further comprises placing the mixed solution in a cool, dry and protected from light, and allowing to stand. The steps of a certain time. Optional, a certain time refers to 2h-24h.

In another embodiment, the treatment of nutritional anemia, hemolytic anemia, and hemorrhagic anemia comprises at least one of the following:

1) The number of abnormal red blood cells caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, such as: increasing the amount of red blood cells produced;

2) The amount of abnormal hemoglobin caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, such as: increasing the amount of hemoglobin produced;

3) The abnormal hematocrit caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, such as: increasing hematocrit;

4) The number of abnormal platelets caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, such as: increasing the amount of platelets produced;

5) The abnormal average red blood cell hemoglobin concentration caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, such as: increasing the average red blood cell hemoglobin concentration;

6) The average volume of abnormal red blood cells caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, such as: increasing the average volume of red blood cells;

7) The number of abnormal white blood cells caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, such as: increasing the production of white blood cells.

In another embodiment, the above-mentioned application, method, pharmaceutical composition, health food or health care product, the complications of nutritional anemia, hemolytic anemia, and hemorrhagic anemia include at least one of the following:

(1) Tissue caused by renal ischemia caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia Damage or impaired function;

(2) tissue damage or impaired function caused by hepatic ischemia caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia;

(3) tissue damage or impaired function caused by spleen ischemia caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia;

(4) tissue damage or impaired function caused by bone marrow ischemia caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia;

(5) Tissue damage or impaired function caused by lung ischemia caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia.

In another embodiment, the above-mentioned application, method, pharmaceutical composition, health food or health care product, the complications of treating nutritional anemia, hemolytic anemia, and hemorrhagic anemia include at least one of the following:

(1) making the number and quality of blood cells in liver tissue normal;

(2) making the number and quality of blood cells in the spleen tissue normal;

(3) making the number and quality of blood cells in the bone marrow normal;

(4) making the number and quality of blood cells in the kidney tissue normal;

(5) The number and quality of blood cells in the lung tissue tend to be normal.

In another embodiment of the above application, method, pharmaceutical composition, health food or health care product, the nutritional anemia may be at least one of iron deficiency anemia and megaloblastic anemia.

In another embodiment, the hemolytic anemia may be at least one of globin chain-forming anemia and immunological hemolytic anemia.

Among them, the specific cause of iron deficiency anemia may be at least one of the following factors: insufficient iron supply in the diet or unreasonable dietary structure, insufficient iron absorption, increased iron demand and insufficient intake. Among them, the iron supply in the diet is not enough or the diet structure is unreasonable, and the iron absorption is insufficient, mainly because the iron content of the food intake is insufficient, the iron supply in the diet is mainly non-heme iron, and the iron provided may not be absorbed. Wait. The demand for iron is increased and the intake is insufficient, mainly due to the decrease of iron content in the blood caused by blood loss, such as gastrointestinal bleeding, excessive menstrual flow in women, hemorrhage, hemoptysis, hematuria, nose bleeding, hookworm disease and chronic digestive tract. Hemorrhage, chronic renal insufficiency, hemodialysis, etc. can lead to iron deficiency anemia. In the above application, the iron deficiency anemia is specifically caused by at least one of the following factors: insufficient iron supply in the diet or an unreasonable dietary structure, insufficient iron absorption, and insufficient intake of iron;

The specific cause of megaloblastic anemia can be at least one of the following factors: vitamin B12 and/or folate deficiency. Among them, the most common cause of vitamin B12 deficiency in foreign countries is the pernicious anemia caused by the decrease of internal factors; the common cause of folic acid deficiency is dietary factors such as chronic alcoholism, drug reaction or malabsorption; other causes of vitamin B12 and folate deficiency include Defects in operation, increased demand (such as pregnancy and milk secretion), malignant tumors, chronic hemolysis, metabolic diseases, and premature babies.

The hemolytic anemia may be at least one of a globin chain-forming anemia (also called thalassemia or marine anemia) and immune hemolytic anemia.

Among them, the specific cause of globin chain-forming anemia can be at least one of the following factors: genetic or genetic mutation. The inheritance may be an abnormal globin synthesis gene inherited by the parent; the mutation may be a physical, chemical, biological or the like induced mutation, specifically X-ray, bacteria, virus, etc. induction.

The specific cause of immune hemolytic anemia can be at least one of the following: chemical substances, physical factors, and biotoxins. Chemical substances mainly include lead poisoning, copper poisoning, hydrogen arsenide, hydrogen telluride, elemental aluminum, aromatic nitro or amino compounds, benzoquinone, drugs, etc. Among them, the drugs mainly include furopyridine, dapson and amino groups. Sulfamides such as salicylic acid, furans, salicylic acids, and aromatic compounds are susceptible to oxidative hemolysis. Physical factors mainly include high temperature, radiation, and hypotonic blood. Biological toxins mainly include snake venom, bee venom, spider toxins and other toxins.

In another embodiment, the medicament or the pharmaceutical composition may be a tablet, a pill, a powder, a lozenge, a sachet, a cachet, an elixir, a suspension, an emulsion, a solution. Formulations of granules, syrups, aerosols, ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions or aseptically packaged powders. In the present invention, the active ingredient is prepared into a pharmaceutical or pharmaceutical composition for immediate release, sustained release or delayed release of the active ingredient after administration to a subject, for example, the active ingredient can be combined with The carrier is mixed, diluted with a carrier or encapsulated in a carrier. The carrier, excipient, and diluent may be either a carrier, an excipient, and a diluent serving as an active ingredient, or a carrier, an excipient, and a diluent serving as a medium.

The medicament or the above pharmaceutical composition in the above application. In another embodiment, suitable carriers, excipients and diluents may be solid, semi-solid or liquid materials such as lactose, dextrose, sucrose, sorbitol , mannitol, starch, resin, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, methyl Cellulose, methyl and propylparaben, talc, magnesium stearate or liquid paraffin.

The medicament or the above pharmaceutical composition in the above application. In another embodiment, the medicament or pharmaceutical composition may additionally comprise a lubricant, a wetting agent, an emulsifying and suspending agent, a preservative, a sweetener or a flavoring agent. .

In another embodiment, when the drug or the pharmaceutical composition is present in a liquid form, the concentration of the active ingredient in the drug or the pharmaceutical composition is 0.01-100 mg/mL, Optionally, it is 0.01-3 mg/mL, 0.01-5 mg/mL, 0.01-10 mg/mL, 0.01-20 mg/mL, 0.01-30 mg/mL, 0.01-40 mg/mL, 0.01-50 mg/mL; when the drug is Or when the pharmaceutical composition is present in a solid form, the concentration of the active ingredient in the drug or the pharmaceutical composition is from 0.01 to 100 mg/g, alternatively from 0.01 to 3 mg/g, from 0.01 to 5 mg/g, 0.01-10 mg/g, 0.01-20 mg/g, 0.01-30 mg/g, 0.01-40 mg/g.

In another embodiment, the subject is a mammal, such as a mouse, a guinea pig, a rat, a dog, a rabbit, a monkey, a human, and the like.

In another embodiment, the active ingredient is administered at a dose of 0.1 mg/kg/d to 100 mg/kg/d, optionally 1-20 mg/kg/d, 1-10 mg/kg/d. Etc., the application course can be 5 days to 30 days, depending on the condition, short-term or long-term administration; the active ingredient can be administered orally, intravenously or intraperitoneally.

In another embodiment, the fullerene structure is used in vivo in a concentration ranging from 0.1 mM to 100 mM. Thereby, the use effect of the drug can be further improved.

As used herein, the term "treatment" includes its generally accepted meaning, which includes preventing, preventing, inhibiting, ameliorating, and slowing, halting, or reversing the development of a symptom or a desired condition. As such, the invention encompasses both therapeutic and prophylactic administration.

The term "effective amount" as used herein, refers to an amount or dose of the active ingredient that is administered to a patient in a single or multiple administrations to provide a desired effect to the patient being diagnosed or treated. The effective amount can be determined by the participating diagnostician as a result of known techniques by those skilled in the art and in similar circumstances. In determining the effective amount or dose of the active ingredient administered, the participating diagnostician should consider a variety of factors including, but not limited to, the mammalian species; volume, age, and general health; the particular disease involved Degree or severity of involvement of the disease; response of the individual patient; specific compound administered; mode of administration; bioavailability properties of the administered formulation; selected dosing regimen; use with drug therapy; Related situation.

As used herein, the terms "raw material fullerene", "fullerene", "fullerene bulk" or other fullerene without modifiers refer to fullerene which has not been modified.

The disclosure of all ranges in the invention should be regarded as a disclosure of all sub-ranges and all point values within the scope. For example, the disclosure of 1-1000 should be considered to also disclose the range of 1-200, 200-300, etc., while also exposing point values of 200, 300, 400, 500, 600, 700, 800, 900, and 100.

Compared with the prior art, the beneficial effects of the present invention are:

(1) The inventors have found through extensive experiments that the fullerene structure not only has a strong ability to scavenge free radicals, but also can be rapidly metabolized in the body, has good biocompatibility, and has little toxic side effects on organisms.

(2) Due to the micro-nano-scale particle size of the fullerene structure and the aggregate size in the aqueous solution, it can be efficiently and rapidly enriched in hematopoietic tissues (liver, spleen, bone marrow, etc.) by blood circulation and osmosis, which is highly efficient. Free radicals, protect cells and hematopoietic tissue, treat nutritional anemia, hemolytic anemia, hemorrhagic anemia, and make the indicators of nutritional anemia, hemolytic anemia, and hemorrhagic anemia normal; the fullerene structure is also nutritious Complications of anemia, hemolytic anemia, and hemorrhagic anemia, such as liver damage in the liver, kidney, and spleen, can improve the number and quality of damaged tissues.

(3) The preparation method of fullerene structure is simple and rapid.

DRAWINGS

1 is a graph showing the results of a test for scavenging free radicals of a water-soluble hydroxylated metal fullerene according to Example 1 of the present invention;

2 shows the effect of water-soluble hydroxylated metal fullerene on the count of red blood cells in the treatment of iron deficiency anemia according to Example 1 of the present invention; the abscissa indicates the number of days of blood taken, and the blood was taken 4 times in the experiment, respectively 5 days (before bleeding, once after drug administration), 8th day and 10th day; the ordinate indicates the amount of change in the red blood cell count of each group relative to the first day of the experiment, and the blood test results for each of the 3 groups were columnar The figure represents the Control group, IDA+GFNCs group and IDA group from left to right;

Figure 3 is a graph showing the effect of water-soluble hydroxylated metal fullerene on hemoglobin content in the treatment of iron deficiency anemia according to Example 1 of the present invention; the abscissa indicates the number of days of blood taken, and the blood was taken 4 times in the experiment, respectively 5 days (before bleeding, once after drug administration), 8th day and 10th day; the ordinate indicates the amount of change of hemoglobin content of each group relative to the first day of the experiment; the blood test results for each of the 3 groups were columnar The figure represents the Control group, IDA+GFNCs group and IDA group from left to right;

Figure 4 is a graph showing the effect of water-soluble hydroxylated metal fullerene on hematocrit in the treatment of iron deficiency anemia according to Example 1 of the present invention; the abscissa indicates the number of days of blood taken, and the blood was taken 4 times in the experiment, respectively Day 5 (before bleeding, once after drug administration), Day 8 and Day 10; ordinate indicates the amount of change in hematocrit of each group relative to day 1 of the experiment; blood test for each group for 3 groups Results The histograms represent the Control group, IDA+GFNCs group and IDA group from left to right;

Figure 5 is a graph showing the effect of water-soluble hydroxylated metal fullerene on platelet in the treatment of iron deficiency anemia according to Example 1 of the present invention; the abscissa indicates the number of days of blood taken, and the blood was taken 4 times in the experiment, respectively. Days (before bloodletting, each time after drug administration), Day 8 and Day 10; ordinate indicates the amount of change of platelets relative to the first day of the experiment; the histogram of blood test results for each of the three groups was from Left to right represent Control group, IDA+GFNCs group and IDA group respectively;

Figure 6 shows a water-soluble hydroxylated metal fullerene according to Example 1 of the present invention in the treatment of iron deficiency The effect of anemia on the mean erythrocyte hemoglobin concentration; the abscissa indicates the number of days of blood collection, and the blood was taken 4 times in the experiment, respectively on the 5th day (before bleeding, once after the fight), 8th and 10th; The coordinates indicate the amount of change in the mean erythrocyte hemoglobin concentration of each group relative to the first day of the experiment; the histogram of the blood sampling results for each of the three groups represents the Control group, the IDA+GFNCs group, and the IDA group from left to right;

7 shows an environmental scanning electron micrograph of a water-soluble hydroxylated metal fullerene according to Embodiment 1 of the present invention for improving liver tissue in the treatment of iron deficiency anemia;

8 shows an environmental scanning electron micrograph of a water-soluble hydroxylated metal fullerene according to Example 1 of the present invention for improving spleen tissue in the treatment of iron deficiency anemia;

9 shows an environmental scanning electron micrograph of a water-soluble hydroxylated metal fullerene according to Embodiment 1 of the present invention for improving bone marrow in the treatment of iron deficiency anemia;

Figure 10 is a diagram showing an environmental scanning electron micrograph of a water-soluble hydroxylated metal fullerene according to Example 1 of the present invention for improving renal tissue in the treatment of iron deficiency anemia;

Figure 11 is a view showing an optical micrograph (400X) of a pathological section of a liver tissue improved by a water-soluble hydroxylated metal fullerene according to Example 1 of the present invention in the treatment of iron deficiency anemia;

Figure 12 shows an optical micrograph (400X) of a pathological section of a water-soluble hydroxylated metal fullerene for improving renal tissue in the treatment of iron deficiency anemia according to Example 1 of the present invention;

Figure 13 shows an optical micrograph (200X) of a pathological section of a water-soluble hydroxylated metal fullerene for improving spleen tissue in the treatment of iron deficiency anemia according to Example 1 of the present invention.

Figure 14 is a graph showing the effect of water-soluble hydroxylated metal fullerenes on the count of red blood cells in the treatment of hemolytic anemia according to Example 1 of the present invention; the abscissa indicates the number of days of blood taken, and the blood was taken 5 times in the experiment, respectively being 0. , 4, 8, 13, 18 days; the ordinate indicates the amount of change in the red blood cell count of each group relative to the 0th day of the experiment, and the histogram of the blood test results for each of the 3 groups represents the Control group from left to right. HA+GFNCs group and HA group;

Figure 15 is a graph showing the effect of water-soluble hydroxylated metal fullerene on hemoglobin content in treating hemolytic anemia according to Example 1 of the present invention; the abscissa indicates the number of days of blood taken, and blood was taken from the experiment. Times, 0, 4, 8, 13, 18 days respectively; the ordinate indicates the amount of change in the red blood cell count of each group relative to the 0th day of the experiment, and the histogram of the blood test results for each of the 3 groups from left to left The right represents the Control group, the HA+GFNCs group, and the HA group, respectively;

Figure 16 is a graph showing the effect of water-soluble hydroxylated metal fullerene on the count of red blood cells in the treatment of hemolytic anemia according to Example 1 of the present invention; the abscissa indicates the number of days of blood taken, and the blood was taken 5 times in the experiment, respectively being 0. , 4, 8, 13, 18 days; the ordinate indicates the amount of change in the hematocrit of each group relative to the 0th day of the experiment, and the histogram of the blood test results for each of the 3 groups represents the Control group from left to right. , HA+GFNCs group and HA group;

Figure 17 is a graph showing the effect of water-soluble hydroxylated metal fullerene on the average volume of red blood cells in treating hemolytic anemia according to Example 1 of the present invention; the abscissa indicates the number of days of blood taken, and the blood was taken 5 times in the experiment, respectively. 0, 4, 8, 13, 18 days; the ordinate indicates the amount of change in the red blood cell count of each group relative to the 0th day of the experiment, and the histogram of the blood sampling results for each of the 3 groups represents the Control group from left to right. , HA+GFNCs group and HA group;

Figure 18 is a diagram showing an environmental scanning electron micrograph (3000X) of water-soluble hydroxylated metal fullerenes in the treatment of hemolytic anemia according to Example 1 of the present invention;

Figure 19 shows an environmental scanning electron micrograph (3000X) of a water-soluble hydroxylated metal fullerene according to Example 1 of the present invention for improving spleen tissue in the treatment of hemolytic anemia;

Figure 20 is a diagram showing an environmental scanning electron micrograph (3000X) of a water-soluble hydroxylated metal fullerene according to Embodiment 1 of the present invention for improving liver in the course of treating hemolytic anemia;

21 shows an environmental scanning electron micrograph (3000X) of a water-soluble hydroxylated metal fullerene according to Embodiment 1 of the present invention for improving renal tissue during treatment of hemolytic anemia;

Figure 22 shows the effect of fullerene olive oil treatment on white blood cell counts in each group. Four histograms per day corresponded to blank control group, fullerene group, anemia group and fullerene treatment from left to right. group;

Figure 23 shows the effect of fullerene olive oil treatment on the red blood cell count of each group of mice. The four histograms per day correspond to the blank control group, the fullerene group, the anemia group and the fullerene from left to right. therapy group;

Figure 24 shows the effect of fullerene olive oil treatment on the hemoglobin content of each group of mice. The four histograms per day corresponded to the blank control group, the fullerene group, the anemia group and the fullerene treatment from left to right. group;

Figure 25 shows the effect of fullerene olive oil treatment on hematocrit in each group of mice. The four histograms per day corresponded to the blank control group, fullerene group, anemia group and Fuller from left to right. Ane treatment group;

Figure 26 shows the effect of fullerene olive oil treatment on the body weight of each group of mice;

Figure 27 shows the effect of fullerene olive oil treatment on the organ coefficient of each group of mice. The four histograms of each organ corresponded to the blank control group, fullerene group, anemia group and Fuller from left to right. Ane treatment group;

Figure 28 shows the effect of fullerene olive oil treatment on the two organ coefficients of each group of mice. The four histograms of each organ corresponded to the blank control group, fullerene group, anemia group and Fuller from left to right. Ane treatment group;

Figure 29 is a 200X, 400X observation of a course of bone marrow pathology in mice treated with fullerene olive oil. From left to right, the blank control group, the fullerene group, the anemia group and the fullerene treatment group;

Figure 30 is a 200X, 400X observation of the treatment of two bone marrow pathology in mice by fullerene olive oil treatment, from left to right, blank control group, fullerene group, anemia group and fullerene treatment group;

Fig. 31 shows the treatment of fullerene olive oil on the course of a bone marrow environment scanning electron microscope 3000X, 10000X in mice, from left to right, blank control group, fullerene group, anemia group and fullerene treatment group;

Figure 32 shows the observation of fullerene olive oil treatment in mice treated with two bone marrow environment scanning electron microscope 3000X, 10000X, from left to right as blank control group, fullerene group, anemia group and fullerene treatment group;

Figure 33 is a 200X observation of a full-range olive oil treatment on a pathological section of a spleen in mice, from left to right, a blank control group, a fullerene group, an anemia group, and a fullerene treatment group;

Figure 34 is a 200X observation of fullerene olive oil treatment on the pathological section of two spleen in mice, from left To the right are the blank control group, the fullerene group, the anemia group and the fullerene treatment group;

Fig. 35 shows the observation of fullerene olive oil treatment on the spleen environment of mice by scanning electron microscopy 3000X, 10000X, from left to right, blank control group, fullerene group, anemia group and fullerene treatment group;

Fig. 36 shows the observation of fullerene olive oil treatment on the spleen environment of mice by scanning electron microscopy 3000X, 10000X, from left to right, blank control group, fullerene group, anemia group and fullerene treatment group;

Figure 37 is a 200X observation of a kidney pathological section of a fullerene olive oil treatment in mice, from left to right, a blank control group, a fullerene group, an anemia group, and a fullerene treatment group;

Fig. 38 is a 200X observation of the pathological section of the two kidneys treated with fullerene olive oil treatment, from left to right, the blank control group, the fullerene group, the anemia group and the fullerene treatment group;

Figure 39 shows the treatment of fullerene olive oil in mice treated with a renal environment scanning electron microscope 3000X, 10000X, from left to right, blank control group, fullerene group, anemia group and fullerene treatment group;

Figure 40 shows the treatment of fullerene olive oil in mice treated with two kidneys scanning electron microscopy 3000X, 10000X, from left to right, blank control group, fullerene group, anemia group and fullerene treatment group;

Figure 41 is a 200X observation of liver pathological section of mice treated with fullerene olive oil treatment, from left to right, blank control group, fullerene group, anemia group and fullerene treatment group;

Figure 42 is a 200X observation of fullerene olive oil treatment on the liver pathological section of mice. From left to right, the blank control group, the fullerene group, the anemia group and the fullerene treatment group;

Fig. 43 shows the treatment of fullerene olive oil on the course of treatment of mice by a liver environment scanning electron microscope 3000X, 10000X, from left to right, respectively, blank control group, fullerene group, anemia group and fullerene treatment group;

Figure 44 shows the observation of fullerene olive oil treatment in mice treated with liver microscopy 3000X, 10000X, left to right, blank control group, fullerene group, anemia group and fullerene treatment group;

Figure 45 is a 200X observation of a pathological section of a lung in mice treated with fullerene olive oil treatment, from left to right, blank control group, fullerene group, anemia group and fullerene treatment group;

Figure 46 is a 200X observation of the pathological section of the lungs treated with fullerene olive oil in the course of treatment. From left to right, the blank control group, the fullerene group, the anemia group and the fullerene treatment group;

Figure 47 shows the treatment of fullerene olive oil on the lungs of the lungs in the lungs of 3000X and 10000X. From left to right, the blank control group, the fullerene group, the anemia group and the fullerene treatment group;

Fig. 48 shows the observation of fullerene olive oil treatment on the lungs of the lungs in the lungs of 3000X and 10000X. From left to right, the blank control group, the fullerene group, the anemia group and the fullerene treatment group.

detailed description

The specific embodiments of the present invention are described in detail below with reference to the accompanying drawings, but it is understood that the scope of the present invention is not limited by the specific embodiments.

The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

The raw material Gd@C ₈₂ solid powder used in the following examples was purchased from Xiamen Funa New Material Technology Co., Ltd., with a molecular weight of 1141 and a purity of 99.1%. The raw material C ₆₀ solid powder used in the following examples was purchased from Xiamen Funa New Material Technology Co., Ltd., with a molecular weight of 720 and a purity of 99%. The low-iron feed and common feed used in the following examples were purchased from Beijing Huakang Biotechnology Co., Ltd., and the iron content in low-iron feed and ordinary feed was determined by ICP-AES (inductively coupled plasma emission spectrometry). It is 20 mg/kg and 200 mg/kg.

Example 1: Preparation of water-soluble hydroxylated metal fullerenes:

1) 100 mg of Gd@C ₈₂ solid powder was added to a 100 ml single-mouth bottle, and 7 ml of a 30% aqueous solution of hydrogen peroxide and 3 ml of a 2 mol/L aqueous solution of sodium hydroxide were added, and the mixture was heated to 70 ° C in an oil bath. 2-5h.

2) After the reaction, the small molecule was removed using a MW=3500 dialysis bag, and monitored by a conductivity meter until the completion of dialysis. The concentrated product was a water-soluble hydroxylated metal fullerene, which was determined by dynamic light scattering (DLS) in an aqueous solution. The average particle diameter was 140 nm, and the particle size distribution was uniform. In the following examples, they are referred to as GFNCs.

Example 2: Preparation of water-soluble hydroxylated hollow fullerenes:

1) Add 100 mg of C ₆₀ solid powder to a 100 ml single-mouth bottle, add 7 ml of a 30% volume aqueous hydrogen peroxide solution and 3 ml of a 2 mol/L sodium hydroxide aqueous solution, and heat to 70 ° C in an oil bath. 5h.

2) After the reaction, the small molecule is removed by using MW=3500 dialysis bag, and monitored by conductivity meter until the completion of dialysis, and the obtained product is concentrated to obtain water-soluble hydroxylated hollow fullerene, and the average particle diameter in the aqueous solution is determined by DLS. It is 143.4 nm and the particle size distribution is uniform.

Example 3: Detection of free radical scavenging ability of water-soluble hydroxylated metal fullerene

The present invention detects the ability of water-soluble hydroxylated ruthenium metal fullerene to scavenge free radicals by electron spin resonance spectroscopy (ESR).

The hydroxyl radical has a short lifetime and cannot be directly measured, so a hydroxyl radical trapping reagent lutidine N-oxide (DMPO) is used. The hydroxyl radical is generated by the ultraviolet-induced method, and then the hydroxy radical generated by DMPO capture generates DMPO radical, while the DMPO radical is relatively stable, and its intensity can be measured by ESR. In the reaction system, as the concentration of hydroxyl radicals increases, the signal intensity of DMPO radicals becomes higher and higher. If hydroxyl radicals are quenched, the formation of DMPO radicals is inhibited. Using this principle, the effect of GFNCs scavenging hydroxyl radicals is indirectly obtained by inhibiting DMPO radicals.

In the test, the control group was: 50 μL of 37% hydrogen peroxide solution, 50 μL PBS buffer (pH=7.4) solution was mixed, and then mixed with 100 μL (0.133 mM) (DMPO, free radical trap) solution, using 280 nm ultraviolet light. Irradiation for 8 min; experimental group: 50 μL mass concentration of 37% Hydrogen peroxide was mixed with 100 μL (0.133 mM) lutidine N-oxide (DMPO, radical scavenger) solution, and immediately added 5 μM of 50 μL of water-soluble hydroxylated ruthenium metal fullerene solution prepared in Example 1 for use. The signal intensity of free radicals was detected by 280 nm ultraviolet light irradiation for 8 min.

As shown in Fig. 1, in the experimental group, when the concentration of the GFNCs in the example 1 was 5 μM, the radical signal was significantly weakened relative to the control group, that is, the free radical generated by the ultraviolet light irradiation of the hydrogen peroxide was effectively quenched.

Example 4, animal experiment for treating iron deficiency anemia

1. Animal models and grouping

Animal model: ICR 9-week-old female mice were randomly divided into 3 groups, 6 in each group, corresponding to Control group, IDA group and IDA+GFNCs group. Among them: Control group was blank control group, IDA group was lacking. In the iron anemia group, the IDA+GFNCs group was a water-soluble hydroxylated ruthenium metal fullerene group administered to a group of subjects with iron deficiency anemia.

Control group: feeding ordinary feed; no bloodletting; replacing the drugs injected by IDA+GFNCs group with the same volume of normal saline, and injecting them into mice by tail vein; other treatments were consistent with IDA+GFNCs group.

IDA group: feeding low-iron diet; bloodletting (0.2-0.3ml/time); replacing the drug injected with IDA+GFNCs group with the same volume of normal saline, and injecting the tail vein into mice; other treatment methods and IDA+GFNCs group Consistent.

IDA+GFNCs group: fed low-iron diet; bloodletting (0.2-0.3 ml/time); the drug was injected intravenously into mice at a dose of 1.005 mM GFNCs per injection of 160 μl.

2, experimental methods

(1) Blood image detection

The mice were cultured for 2-3 days, the state of the mice was observed, and relevant indicators of the mice, such as red blood cells (RBC), hemoglobin (HGB), platelet (PLT) hematocrit (HCT), and mean erythrocyte hemoglobin concentration (MCHC) were measured. ), wherein: RBC is a red blood cell count, red blood cells are also called red blood cells, which is the most abundant blood cell in the blood, mainly produced in the bone marrow, and functions as transport oxygen. Qi, participate in immunity, etc.; red blood cell count is lower than normal, common in: blood loss, anemia, hemorrhage, bone marrow hematopoietic disorders, leukemia, malnutrition, erythropoietin deficiency. HGB is the hemoglobin content. Hemoglobin is the main component of red blood cells. It is a protein that makes blood red, and a protein that transports oxygen in higher organisms. It is also the main reference standard for diagnosis and treatment of anemia. PLT is a platelet derived from cytoplasmic granules of bone marrow megakaryocytes. Its main function is to participate in hemostasis and coagulation, promote endothelial cell proliferation, repair blood vessels, etc.; causes thrombocytopenia: bone marrow hematopoietic dysfunction, aplastic anemia, etc. In addition, poisoning or allergies caused by taking some drugs will also reduce thrombocytopenia. HCT is hematocrit, which refers to the percentage of red blood cells in the volume of whole blood, reflecting the ratio of red blood cells to plasma. MCHC is the average erythrocyte hemoglobin concentration, MCHC=HGB/HCT, which is generally used as an auxiliary standard for the diagnosis and treatment of anemia and related diseases, and is helpful for analyzing the causes and treatment of anemia.

After the above data was stabilized, it was the first day of the experiment. Then, on the first day and the fifth day (AM), blood was taken from the eyelids of the mice, and GFNCs were injected into the mice on the 5th (afternoon), 6th, and 7th days, respectively, on the 5th day. (Before bleeding, after each fight), on the 8th day and the 10th day, take 20μl of blood from the eye of the mouse. As for the 3ml centrifuge tube, use the pre-dilution mode of the automatic analyzer to detect anemia-related indicators: RBC, HGB, PLT, HCT and MCHC, the test results are shown in Figure 2-6, respectively (the difference between each indicator and the starting value of each group is shown in the figure).

(2) Improve liver, spleen and kidney during the treatment of iron deficiency anemia

On the 11th day, 2 mice were randomly selected in the control group, IDA group, IDA+GFNCs group, and sacrificed by cervical dislocation. The liver, spleen, bone and kidney were taken and labeled.

Environmental scanning electron microscope sample preparation: (1) fixation: 2.5% glutaraldehyde fixed liver, spleen, bone, kidney samples for more than 4h, then washed with phosphate buffer 3 times, each time 15 ~ 20min (2) dehydration: 30%, 50%, 70%, 85%, 95% ethanol, each time 15 to 20 minutes. 100% ethanol 2 times, each time 15 ~ 20min (3) replacement: isoamyl acetate 2 times, each time 15min or more (4) CO2 critical point drying. The FEG field emission environment scanning electron microscope (using a low vacuum mode) was used to observe, select a suitable area, and photograph. Select at least 4 areas for each sample to observe and contrast.

Pathological section observation sample preparation: liver, spleen and kidney samples were fixed with 4% formaldehyde and sent to Peking University Medicine The samples were prepared and then observed under a 400X microscope.

3. Experimental results

(1) Blood test results

As can be seen from Fig. 2, Fig. 3 and Fig. 4, the RBC, HGB and HCT indexes of the IDA+GFNCs group and the IDA group were significantly reduced after the bloodletting, and the anemia standard was reached; the IDA+GFNCs group obtained the blood obtained after the drug was beaten on the fifth day. No change or slight decrease, the increase in blood collection index on the 10th day is very obvious, which is closer to the Control group; while the blood collection index on the 8th day of the IDA group is still significantly lower, the blood gain index on the 10th day is slightly Recovery, but the magnitude is small, far less than the results of the IDA+GFNCs group.

This may be due to the lack of blood cells in the blood of the IDA group, and insufficient iron supplementation, the hematopoietic tissue can not play a role in time, resulting in iron deficiency anemia; stop bleeding, the body still shows anemia symptoms for a period of time, although after It can be restored by its own adjustment, but the period is longer and the amplitude is smaller. The injection of GFNCs into the IDA+GFNCs group can promote the related role of hematopoietic tissue and improve the utilization of iron. In the short term, it can properly compensate for iron deficiency caused by blood loss. After a period of time, the effect is more obvious, and it is related to anemia. The indicators have basically returned to normal. This may be due to the fact that GFNCs can improve the utilization of iron elements to a certain extent, improve the role of hematopoietic tissue, effectively remove hydrogen peroxide from the body, and moderately compensate for the loss of iron.

As can be seen from Figure 5, after bloodletting, PLT showed a downward trend, and PLT increased after injection of GFNCs.

It can be seen from Fig. 6 that the MCHC index is improved after the injection of GFNCs, indicating that the drug can improve the related effects of hematopoietic tissue in mice.

(2) Environmental scanning electron microscopy and pathological section results

The results obtained by environmental scanning electron microscopy are shown in Figures 7-10. As can be seen from the figure, in the main hematopoietic tissues and / or organs: liver (Figure 7), spleen (Figure 8), bone marrow (Figure 9), kidney (Figure 10) samples, the control group of mice with normal blood cells and blood cells Fullness; the number of blood cells in the IDA group (pointed by the arrow) was significantly reduced, and non-blood cells such as fat were increased, while the blood cells in the IDA+GFNCs group (pointed by the arrow) were improved compared with the IDA group, indicating that GFNCs can be somewhat Improve iron deficiency anemia Caused by damage in the liver, spleen, bone marrow, and kidney.

The liver, spleen and kidney samples were observed under a 400X microscope. The results are shown in Figures 11 to 13. The number and quality of blood cells in the liver, spleen and kidney of the IDA+GFNCs group were significantly better than those of the IDA group and were closer to the Control group.

In summary, after injection of water-soluble hydroxylated ruthenium metal fullerene, the number of blood cells in the hematopoietic tissue of mice increased, and the quality improved. Compared with the control group, the water-soluble hydroxylated ruthenium metal fullerene showed iron deficiency. Anemia has a good therapeutic and recovery effect. At the same time, after the blood was released, the liver of the mice was damaged to some extent. After the injection of the water-soluble hydroxylated ruthenium metal fullerene, the liver of the mice was obviously improved. Compared with the blank group, the water-soluble hydroxylated ruthenium metal fuller was observed. The ene has a good protective effect on the liver.

Example 5, animal experiment for treating hemolytic anemia

1. Animal models and grouping

Animal model: ICR 6-8 week old female mice were randomly divided into 3 groups, 6 in each group, corresponding to Control group, HA group and HA+GFNCs group. Among them: Control group was blank control group, HA group In the benzoquinone-induced hemolytic anemia group, the HA+GFNCs group was a water-soluble hydroxylated ruthenium metal fullerene group administered to a group of hemolytic anemia.

Control group: The same volume of physiological saline was used instead of the drug injected in the HA group, and injected intraperitoneally into the mouse.

HA group: The total dosage of benzoquinone is 0.03-0.40g/kg*bw. A total of 3 intraperitoneal injections were performed on

days

1, 4 and 7 of the trial. The first dose was 1/2 of the total dose, and half of the dose was administered on days 4 and 7.

HA+GFNCs group: The injection method of phenylhydrazine was consistent with that of HA group. GFNCs were injected on days 9, 13, and 16. The specific dose was: 1 mM GFNCs were injected 150 μl each time.

2, experimental methods

The mice were cultured for 2-3 days, the state of the mice was observed, and relevant indicators of the mice, such as red blood cells (RBC), hemoglobin (HGB), hematocrit (HCT), and mean red blood cell volume (MCV), including: RBC, were measured. For red blood cell count, red blood cells, also known as red blood cells, are the most abundant type of blood. Blood cells, mainly produced in the bone marrow, function to transport oxygen, participate in immunity, etc.; red blood cell count is lower than normal, common in: blood loss, anemia, hemorrhage, bone marrow hematopoietic disorders, leukemia, malnutrition, erythropoietin deficiency. HGB is the hemoglobin content. Hemoglobin is the main component of red blood cells. It is a protein that makes blood red, and a protein that transports oxygen in higher organisms. It is also the main reference standard for diagnosis and treatment of anemia. HCT is hematocrit, which refers to the percentage of red blood cells in the volume of whole blood, reflecting the ratio of red blood cells to plasma. MCV is the mean red blood cell volume, MCV=HCT/RBC(L)*10^15(fl), which is generally used as an auxiliary standard for the diagnosis and treatment of anemia and related diseases, and is helpful for analyzing the causes and treatment of anemia.

After the above data was stabilized, it was the 0th day of the experiment. Then each group of processing methods are as described above. And on the 0th, 4th, 8th, 13th and 18th day, 20μl of mouse eye blood was taken separately. As for the 3ml centrifuge tube, the anemia-related indicators were detected by the pre-dilution mode of blood cell automatic analyzer: RBC, HGB, HCT and MCV. The results are shown in Figures 14-17 (the differences between the indicators and the starting values of each group are shown in the figure).

On the 19th day, 3 mice were randomly selected in the control group, HA group and HA+GFNCs group, and sacrificed by cervical dislocation. The liver, spleen, bone and kidney were taken and labeled.

The environmental scanning electron microscope sample preparation method is the same as the iron deficiency anemia model.

3. Experimental results

(1) Blood test results

As can be seen from Fig. 14, Fig. 15, Fig. 16 and Fig. 17, respectively, the RBC, HGB, HCT and MCV indexes of HA+GFNCs group and HA group were significantly decreased after injection of phenylhydrazine, and the anemia standard was reached; HA+GFNCs group was injected with GFNCs. The increase in blood collection index was very obvious, which was closer to the Control group. However, the index obtained by the HA group after stopping the injection of phenylhydrazine slightly recovered, but the extent was small, far less than the results of the HA+GFNCs group.

It has been shown that benzoquinone can act on the body's red blood cells, causing hemolysis in a variety of animals. The hemolytic anemia was induced in mice by using phenylhydrazine to establish an acute hemolytic anemia model. As a hemolytic agent, benzoquinone has a fast onset, strong effect, good repeatability, and the intensity of action increases with the injection dose. specialty. After entering the mouse, phenylhydrazine acts on the erythrocyte membrane, accelerating the hydrolysis of leucine, lysine and histidine on the membrane surface, causing a large number of red blood cells to be rapidly destroyed. At the same time, benzoquinone selectively oxidizes the membrane skeleton and globin, and translocates phosphatidylserine to the surface of red blood cells, resulting in reduced red blood cell deformability and enhanced ability of red blood cells to adhere to the extracellular matrix. The above effects are superimposed on each other, so that the destruction rate of red blood cells is much greater than the ability of the body to regenerate red blood cells, resulting in hemolytic anemia in the body.

Therefore, after injection of phenylhydrazine, the blood indexes of HA+GFNCs group and HA group decreased, and after injection of GFNCs, the indexes of HA+GFNCs group were basically consistent with that of Control group, indicating that GFNCs can improve red blood cells and other items. Blood indicators. At the same time, in order to further observe the effects of GFNCs on blood cells and various tissues and organs, an environmental scanning electron microscope was used for observation.

(2) Environmental scanning electron microscope results

The results obtained by environmental scanning electron microscopy are shown in Figures 18-21. As can be seen from the figure, in the main hematopoietic tissues and / or organs: bone marrow (Figure 18), spleen (Figure 19), liver (Figure 20), kidney (Figure 21) samples, the control group of mice with normal blood cells and blood cells Fullness; the number of blood cells in the HA group (pointed by the arrow) was significantly reduced, and non-blood cells such as fat increased, while the blood cells in the HA+GFNCs group (pointed by the arrow) were improved compared with the HA group, indicating that GFNCs can be somewhat Improve the shape and quantity of blood cells in hemolytic anemia, and thus improve the damage of bone marrow, spleen, liver and kidney caused by chemical drugs.

In summary, after injection of water-soluble hydroxylated ruthenium metal fullerene, the blood indexes of the mice were improved, the number of blood cells in the hematopoietic tissue was increased, and the quality was improved. Compared with the control group, water-soluble hydroxylation was observed. Base metal fullerenes have a good therapeutic and recovery effect on hemolytic anemia.

Example 6 Preparation of Fullerene Olive Oil Composition

Take 20ml of olive oil, weigh 20mg C ₆₀ , mix and mix evenly, then put the mixture into the ball mill for ball milling, 10h, after the ball mill is finished, take the mixture out, store it in cool and dry, avoid it, let it stand for a certain period of time, then centrifuge and filter. A fullerene-olive oil solution, which is hereinafter referred to as fullerene olive oil, in which the fullerene content is 0.8 mg/mL, is obtained.

Example 7 Effect of Fullerene Olive Oil Composition in Treating Hemorrhagic Anemia in Animal Experiments

1. Animal models and grouping

Animal models: Female mice with ICR 7-8 weeks old were randomly divided into 4 groups, 10 in each group, corresponding to the blank control group, fullerene group, anemia group and fullerene treatment group.

The blank control group: feeding normal feed; no bloodletting; replacing the drug injected with the fullerene treatment group with the same volume of physiological saline, and intragastrically injecting into the mouse; other treatments were consistent with the fullerene treatment group.

Fullerene group: feeding normal feed; no bloodletting; the fullerene olive oil obtained in Example 6 was intragastrically injected into mice, and the specific dose was 100 μl/time, and the injection time was consistent with the fullerene treatment group.

Anemia group: feeding ordinary feed; bleeding (0.2 ~ 0.3ml / time); using the same volume of normal saline instead of fullerene treatment group of drugs, intragastric injection into mice; other treatment with fullerene treatment The group is consistent.

Fullerene treatment group: feeding normal feed; bleeding (0.2-0.3 ml/time); and fullerene olive oil obtained in Example 6 by intragastric administration in mice, the specific dose was 100 μl/time.

2, experimental methods

(1) Blood image detection

The mice were cultured for 2 to 3 days, and the state of the mice was observed. The relevant indicators of the mice, such as white blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), etc., and WBCs were white blood cells, also called white blood cells, derived from bone marrow. The hematopoietic stem cells in the bone marrow develop into the blood and lymph circulation in the bone marrow, and also exist in the tissues outside the blood vessels and lymphatic vessels. The function is mainly to protect the body from pathogens (ie bacteria and viruses), cancer cells, foreign bodies, etc. . The white blood cell count is lower than normal, and is common in: bone marrow hematopoietic disorders, cytotoxic reactions, autoimmune diseases, liver and spleen diseases, and the like. The role of other indicators has been mentioned in Example 4.

2, experimental methods

(1) Blood image detection

After the indicators were stabilized, as the 0th day of the experiment, the first course of treatment was started after the initial value was collected.

The first course of treatment: bleeding on the eyelids of the mice on the first day, the fourth day, and the seventh day, respectively, and the fullerene olives prepared in Example 6 were intragastrically injected on the 8th, 9th, and 10th days. The oil was in the fullerene-treated group, and 20 μl of the mouse eyelid blood was taken on the 0th, 7th, 11th, and 14th day respectively. As for the 3ml centrifuge tube, the pre-dilution with the blood cell automatic analyzer was performed. Model detection of anemia related indicators: WBC, RBC, HGB, HCT.

The mice were continued to be cultured, and after the indicators were gradually restored and/or stabilized, the second course of the experiment was started.

The second course of treatment: on the 15th, 17th, and 19th day, the blood was bled by the eyelids of the mice, and the fullerene olive oil was administered to the fullerene treatment group on the 20th, 21st, and 22nd days. In mice, 20 μl of mouse eye blood was taken on the 17th, 19th, 22nd, and 26th day respectively. As for the 3ml centrifuge tube, the anemia-related indicator was detected by the pre-dilution mode of the blood cell automatic analyzer: WBC , RBC, HGB, HCT.

(2) Weight detection

The body weight of the mice was weighed on

days

0, 4, 7, 11, 14, 22, and 25.

3. Experimental results

The blood test results of the first course and the second course are shown in Fig. 22 to Fig. 25, respectively, and the body weight test results are shown in Fig. 26.

As can be seen from Figures 22, 23, 24, and 25, after the blood was released, the mice all reached the anemia standard on the 7th day of the first course of treatment, while the mice injected with fullerene olive oil showed various indexes. The recovery is faster, indicating that the fullerene olive oil has a certain therapeutic effect; on the 17th day and the 19th day of the second course of treatment, the anemia group mice are in an anemia standard state, each index is lower, and the fullerene treatment group mice All indicators were slightly higher, probably because of the presence of a certain amount of fullerene olive oil in the mice after the first course of treatment. At the same time, after the injection of fullerene olive oil, the increase was more obvious, basically close to the control group, indicating that the rich Alcene olive oil has a good therapeutic effect on anemia.

As can be seen from Fig. 26, the body weight fluctuation range of each group was basically the same, indicating that the drug has no toxic side effects on mice.

Example 8 Effect of the fullerene olive oil composition on the treatment of anemia and its complications at the tissue level

From Example 7, on the 15th day and the 26th day, 3 mice were randomly selected in the blank control group, the fullerene group, the anemia group and the fullerene treatment group, and the cervical vertebrae were removed by dislocation, and the femur and spleen were taken respectively. Kidney, liver, heart, lung, part of it was fixed with 2.5% glutaraldehyde, and some were fixed with 4% formaldehyde and labeled.

Among them, the organ coefficient of the heart, liver, spleen, lung and kidney of each group were calculated. The organ coefficient is the ratio of an organ of the experimental animal to its body weight. This index is a commonly used index in toxicological experiments. The results are shown in Figure 27. As shown in Fig. 28, the organ coefficients of the two courses of treatment did not change much, indicating that the drug had no obvious side effects. At the same time, the organ coefficient of the anemia group was slightly lower than that of the other groups, and the anemia may have certain damage to the organs.

Pathological section observation sample preparation: fixed with 4% formaldehyde, sent to biophysical preparation, and then observed under a 200X, 400X microscope. As shown in Figures 29, 30, 37, 38, 41, 42, 45, 46.

Environmental scanning electron microscope sample preparation: (1) fixation: 2.5% glutaraldehyde fixed sample for more than 4h, then washed with phosphate buffer 3 times, each time 15 ~ 20min (2) dehydration: 30%, 50%, 70%, 85 %, 95% ethanol each time, each time 15 ~ 20min. 100% ethanol 2 times, each time 15 ~ 20min (3) replacement: isoamyl acetate 2 times, each time 15min or more (4) CO2 critical point drying.

After the sample is prepared, the FEG field emission environment scanning electron microscope (using the low vacuum mode) is used to observe, select a suitable area, and photograph. Select at least 4 areas for each sample to observe and contrast. As shown in Figures 31, 32, 35, 36, 39, 40, 43, 44, 47, 48.

As can be seen from Figures 29, 30, 31, and 32, in the two courses of treatment, as indicated by the arrows, the red pulp of the anemia group was replaced by non-hematopoietic tissues such as white pulp at different degrees, blood cells were significantly reduced, and non-blood cells such as fat were increased. Both the fullerene treatment group improved, indicating that fullerene olive oil can improve the related role of hematopoietic tissue to a certain extent, protect and / or restore the adverse effects of blood loss, and have obvious therapeutic effect on anemia. At the same time, the fullerene group in the two courses was basically consistent with the blank control group, indicating that the fullerene olive oil had no toxic side effects on the bone marrow of mice.

As can be seen from Figures 33 and 34, the degree of damage to the spleen of the mice was gradually increased by two courses of treatment, and the spleen of the mice was significantly improved after the injection of fullerene olive oil, which was closer to the control group, especially in the anemia group. The spleen sinus (as indicated in the figure) was clearly dispersed, and there were more white pulp, while the fullerene treatment group improved. It can be seen that fullerene olive oil has a good protective effect on the spleen. It can be seen from Fig. 35 and 36 that the blood cells in the spleen of the anemia group are reduced or damaged, and the red blood cells are flat, indicating that the bloodletting has a great influence on the blood cell number and quality of the mouse spleen, and the spleen of the fullerene treatment group is spleen. The number and quality of blood cells were significantly better than those of the anemia group. The fullerene group was basically consistent with the blank control group, indicating that the fullerene olive oil had no toxic side effects on the spleen of mice.

As can be seen from Figures 37 and 38, the two courses of treatment, the degree of damage to the kidneys of the mice increased gradually, and the glomeruli were severely damaged. After the injection of fullerenes, the kidneys of the mice were significantly improved. The results of glomerular glomeruli were closer to the blank control group, and it was found that fullerene olive oil had a better protective effect on the kidney. Because the microvascular plexus of the kidney is very rich, the energy demand is particularly high, so bloodletting will cause a certain degree of tissue damage, and ischemia may also have some stimulating effects on physiological responses, and also activate some nerve triggering factors, hormones or drugs. In this experiment, it may be that the fullerene is promoted to scavenge free radicals, thus protecting the kidneys. As shown in Figures 39 and 40, the number of blood cells in the kidney of the anemia group was significantly reduced, and the red blood cells were flat, indicating that the bloodletting had a great influence on the blood cell number and quality of the mouse kidney, while the fullerene treatment group mice. The number and quality of blood cells in the kidneys were significantly better than those in the anemia group. The fullerene group and the blank control group showed basically the same performance, indicating that the fullerene olive oil has no toxic side effects on the kidney of mice.

It can be seen from Fig. 41, 42, 43, and 44 that the degree of damage to the liver of the mice is gradually increased in two courses of treatment, and the anemia group is more severely damaged, and the hepatocytes are atrophied and deformed, and after the fullerene olive oil is injected, There is a modest improvement in the liver of mice, and it can be seen that fullerene olive oil has a good protective effect on the liver. The fullerene group showed little difference from the blank control group, indicating that the fullerene olive oil had no toxic side effects on the liver of mice.

As can be seen from Figures 45, 46, 47, and 48, after two courses of treatment, bloodletting has different degrees of damage to the lungs of the mice, the alveoli are large or small, and there is a certain degree of congestion, and after the injection of fullerene olive oil, There is a modest improvement in the lungs of mice, and it can be seen that fullerene olive oil may have a better protective effect on the lungs. Rich The lenidine group showed little difference from the blank control group, and all of them were composed of relatively homogeneous alveoli, indicating that fullerene olive oil had no toxic side effects on the lungs of mice.

In summary, fullerene olive oil has a significant therapeutic effect on anemia, and has good protection and/or therapeutic effects on bone marrow, spleen, kidney, liver and lung. In addition, this material has essentially no toxic side effects.

The foregoing description of the specific exemplary embodiments of the present invention has The description is not intended to limit the invention to the precise forms disclosed. The embodiments were chosen and described in order to explain the particular embodiments of the invention Choose and change. The scope of the invention is intended to be defined by the claims and their equivalents.

Industrial applicability

The present invention provides a use of a fullerene structure for preparing a medicament for treating anemia and a complication thereof, the fullerene structure comprising a fullerene, a water-soluble modification of the fullerene, the Fuller The olefin is dispersed in at least one of a fullerene composition prepared in an oil solution and a derivative of the fullerene. The fullerene structure can be efficiently and rapidly enriched in hematopoietic tissues (liver, spleen, bone marrow, etc.) through blood circulation and osmosis, which can efficiently scavenge free radicals, protect cells and hematopoietic tissues, and treat nutritional anemia. Hemolytic anemia and hemorrhagic anemia, the indicators of patients with nutritional anemia, hemolytic anemia and hemorrhagic anemia tend to be normal; fullerene structure also complications of nutritional anemia, hemolytic anemia and hemorrhagic anemia, such as The tissue damage of the liver, lung, kidney and spleen caused by the two has a therapeutic effect and can improve the number and quality of the damaged tissue.

Claims

Use of a fullerene structure in the preparation of a pharmaceutical composition, a health care product or a health food, the fullerene structure comprising a fullerene, a water-soluble modification of the fullerene, the fullerene dispersed in an oil solution At least one of a fullerene composition prepared in the form and a derivative of the fullerene for treating nutritional anemia, hemolytic anemia, hemorrhagic anemia, and complications of the above three.
A pharmaceutical composition, health care product or health food for treating nutritional anemia, hemolytic anemia, blood loss anemia and the above three complications, the pharmaceutical composition, the health care product or the health food product including fullerene a structure as an active component, the fullerene structure comprising a fullerene, a water-soluble modification of the fullerene, a fullerene composition prepared by dispersing the fullerene in an oil solution, and the rich At least one of derivatives of olefin.
The pharmaceutical composition according to claim 1, the health care product or the health food according to claim 2, wherein the fullerene comprises at least one of hollow fullerenes and embedded metal fullerenes Kind, of which:

The hollow fullerene is one or more cage structures composed of carbon atoms of the formula C 2m , 20 ≤ m ≤ 60 , optionally C 60 , C 70 , C 84 ;

The inlaid metal fullerene includes M@C 2n , M 2 @C 2n , MA@C 2n , M 3 N@C 2n , M 2 C 2 @C 2n , M 2 S@C 2n , M 2 O One or more of @C 2n and M x A 3-x N@C 2n , wherein: M and A each represent a metal element and M and A are each selected from any one of Sc, Y and a lanthanide metal element. Species, 20≤n≤60; 0≤x≤3; optional Gd@C 82 , Gd@C 60 .
The pharmaceutical composition according to claim 3, the health care product or the health food according to claim 3, wherein the water-soluble modification of the fullerene comprises one or more selected from the group consisting of Fullerenes: (1) fullerenes having a hydrophilic group modified on the surface; (2) fullerenes encapsulated by hydrophilic biological small molecules; (3) rich by biocompatible carrier materials (4) a water-soluble supramolecular system fullerene formed by self-assembly.
The use according to claim 4, the pharmaceutical composition according to claim 4, and a health supplement Or a health food product, wherein the hydrophilic group comprises one or more of a hydroxyl group, a carboxyl group, a thiol group, an amino group, and a water-soluble amino acid residue; optionally, the water-soluble amino acid residue is alanine At least one of a residue, a glycine residue, a serine residue, an arginine residue, a lysine residue, and a tyrosine residue.
The pharmaceutical composition according to claim 5, the health care product or the health food according to claim 5, wherein the water-soluble modification of the fullerene is a water-soluble hydroxylated ruthenium metal fullerene, Water-soluble hydroxylated C 60 , water-soluble hydroxylated C 70 , water-soluble aminated ruthenium metal fullerene, optionally Gd@C 82 (NH 2 ) n , water-soluble aminated C 60 , water-soluble aminated C 70 The water-soluble carboxylated ruthenium metal fullerene may be selected from Gd@C 82 (COOH) n , water-soluble carboxylated C 60 , water-soluble carboxylated C 70 , etc., and further optional water solubility of the fullerene The modification is of the formula C 2a (OH) b or M@C 2d (OH) e , wherein: 20 ≤ a ≤ 60, optionally a is 30 or 35; 0 < b < 50, optional 0 < b <30, optionally also b=13, 20, 24; M is selected from the group consisting of rare earth metals, the optional rare earth metals are Gd, La; 20 ≤ d ≤ 60, and the optional d is 41 or 30 or 35; e<50, optional 0<e<30, and optional e=13, 20, 24.
The use according to claim 5, the pharmaceutical composition, the health care product or the health food according to claim 5, wherein the method for preparing fullerene having a surface modified with a hydroxyl group comprises directly contacting the fullerene body with H 2 The O 2 is obtained by reacting under alkaline conditions. The preparation method further comprises: (a) adding an aqueous hydrogen peroxide solution (optionally, the aqueous hydrogen peroxide solution is 1-30% by mass) and a sodium hydroxide solution. / Potassium hydroxide solution (optional, sodium hydroxide solution / potassium hydroxide solution in a mass percentage of 10-80%) mixed (optional, according to aqueous hydrogen peroxide solution and sodium hydroxide solution / potassium hydroxide The volume ratio of the solution is 1-10:1), and the fullerene is added to the mixture (optionally, 20-500 mg of fullerene is added per 10-200 ml of the mixture) at a temperature of 50-80 ° C. Reaction (optionally, the reaction is stirred for 4-24 h; optionally, stirring speed is 1000 r/min), filtered, and the filtrate is retained; (b) the filtrate is added to excess ethanol (optional, The ethanol concentration is 85%-100%), after centrifugation (optional, the centrifugal speed is 10000r/min, the centrifugation time is The precipitate was collected after 1-10 min), the precipitate was dissolved in water to obtain a solution; (c) the solution obtained in the step (b) was subjected to dialysis treatment.
The use according to claim 1, the pharmaceutical composition according to claim 2, and a health supplement Or a health food product, characterized in that the fullerene structure has an average hydrated particle diameter of from 1 to 1000 nm, and optionally, such as an average hydrated particle diameter of from 1 to 200 nm.
The pharmaceutical composition, health food or health food according to claim 1, wherein the fullerene is dispersed in an oil solution by using fullerene and an oil solution. After the mixture is subjected to ball milling or ultrasonication, the precipitate is removed by centrifugation in sequence, and then the obtained supernatant liquid is filtered to remove particles, that is, alternatively, the oil solution may be a single component oil or a mixture of different oils. Oil; usually vegetable oil, such as olive oil, linseed oil, sunflower oil, corn germ oil, soybean oil, etc., also includes animal fats such as squalane.
The pharmaceutical composition, health food or health food according to claim 1, wherein the fullerene composition has a concentration of fullerene of 0.01 to 100 mg/mL, optionally 0.01-0.8 mg/mL, 0.01-1 mg/mL, 0.01-10 mg/mL, 10-20 mg/mL, 20-30 mg/mL, 30-40 mg/mL, and the like.
The use according to claim 9, wherein the pharmaceutical composition, the health care product or the health food according to claim 9, wherein 0.05-1000 mg is mixed per 1 ml of the oil solution during mixing of the fullerene and the oil solution. Fullerenes, optionally 0.05-1 mg, 0.05-10 mg, 0.05-100 mg, and the like.
The pharmaceutical composition according to claim 9, the health care product or the health food according to claim 9, wherein the mixed solution is ball milled or ultrasonic for 30 min to 15 h, optionally 8-12 h, 10 h. .
The pharmaceutical composition, health care product or health food according to claim 1, wherein the therapeutic nutritional anemia, hemolytic anemia, and hemorrhagic anemia comprise at least one of the following:

1) The number of abnormal red blood cells caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, and the amount of red blood cells produced is optional;

2) The amount of abnormal hemoglobin caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, and the amount of hemoglobin produced may be increased;

3) The abnormal hematocrit caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to Normal, optional to increase hematocrit;

4) The number of abnormal platelets caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, and the amount of platelet production can be increased;

5) The abnormal average erythrocyte hemoglobin concentration caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, and the average erythrocyte hemoglobin concentration may be increased;

6) The average volume of abnormal red blood cells caused by nutritional anemia, hemolytic anemia and hemorrhagic anemia tends to be normal, and the average volume of red blood cells can be increased;

7) The number of abnormal white blood cells caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia tends to be normal, such as: increasing the production of white blood cells;

Optionally, the nutritional anemia is at least one of iron deficiency anemia and megaloblastic anemia, and the hemolytic anemia is at least one of globin chain-forming anemia and immune hemolytic anemia. .
The pharmaceutical composition, health care product or health food according to claim 1, wherein the complications of nutritional anemia, hemolytic anemia, and hemorrhagic anemia include at least one of the following:

(1) tissue damage or impaired function caused by renal ischemia caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia, optionally, the number of blood cells in the renal tissue is reduced, and the blood cells are flattened;

(2) tissue damage or impaired function caused by hepatic ischemia caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia, optionally, the number of blood cells in the liver tissue is reduced, and the blood cells are flattened;

(3) tissue damage or impaired function caused by spleen ischemia caused by nutritional anemia, hemolytic anemia and hemorrhagic anemia, optionally, the number of blood cells in the spleen tissue is reduced, and the blood cells are flattened;

(4) tissue damage or impaired function caused by bone marrow ischemia caused by nutritional anemia, hemolytic anemia and hemorrhagic anemia, optionally, the number of blood cells in the bone marrow is reduced, and the blood cells are flattened;

(5) Tissue damage or impaired function caused by lung ischemia caused by nutritional anemia, hemolytic anemia, and hemorrhagic anemia.
The pharmaceutical composition, health care product or health food according to claim 1, wherein the complication of treating nutritional anemia, hemolytic anemia, and hemorrhagic anemia comprises at least one of the following :

(1) making the number and quality of blood cells in liver tissue normal;

(2) making the number and quality of blood cells in the spleen tissue normal;

(3) making the number and quality of blood cells in the bone marrow normal;

(4) making the number and quality of blood cells in the kidney tissue normal;

(5) The number and quality of blood cells in the lung tissue tend to be normal.