CN109925322B - Application of water-soluble fullerene structure in preparation of medicine for treating pancreatic diseases - Google Patents
Application of water-soluble fullerene structure in preparation of medicine for treating pancreatic diseases Download PDFInfo
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Abstract
The embodiment of the invention relates to application of a water-soluble fullerene structure in preparing a medicament for treating pancreatic diseases, wherein the water-soluble fullerene structure comprises at least one of the following components: a water-soluble modified hollow fullerene, a water-soluble modified endohedral metallofullerene, a combination of the water-soluble modified hollow fullerene and the water-soluble modified endohedral metallofullerene, pharmaceutically acceptable esters thereof or pharmaceutically acceptable salts thereof; the pancreatic diseases include pancreatic cancer, pancreatitis and pancreatic cysts. The water-soluble fullerene structure of the effective component can be selectively enriched at the pancreatic part and treat pancreatic diseases such as pancreatitis and pancreatic cancer in a relatively short time.
Description
Technical Field
The invention relates to the field of biological medicine, in particular to application of a water-soluble fullerene structure in preparation of a medicine for treating pancreatic diseases.
Background
The pancreas, second only to the liver, is the 2 nd largest gland in the human body, which is divided into two parts, exocrine and endocrine glands. Wherein: the exocrine gland consists of acinus and ducts, the acinus is a cell secreting pancreatic juice, and the pancreatic juice secreted by the acinus is discharged into duodenum through the duct of the pancreatic gland, so that the acinus has the functions of neutralizing gastric acid and digesting protein, fat and sugar; endocrine glands are composed of islet cell masses of varying sizes, and regulate blood glucose and participate in metabolism in the body by secreting insulin and glucagon.
The pancreas, although small, has extraordinary effects, and its physiological effects and pathological changes are closely related to life. Therefore, the pathological changes of the pancreas pose a great threat to the human life and health. Major diseases of the pancreas include, but are not limited to, pancreatic cancer, pancreatitis, and the like. Pancreatic cancer is a malignant tumor of the digestive tract with high malignancy and difficulty in diagnosis and treatment. Pancreatitis includes acute pancreatitis, chronic pancreatitis and complications such as pancreatic cyst, pancreatitis is caused by obstructed pancreatic and pancreatic outflow or unsmooth excretion, bile reflux into pancreatic tissue, biliary tract infection or bacterial action, etc. The common pancreatic diseases are serious in disease symptoms, difficult to cure radically, and harmful to human bodies to different degrees, and are the hot spots of current research.
Fullerene is another allotrope of carbon elements other than graphite, diamond, and amorphous carbon. Such substances are referred to as cage-like structures consisting of carbon atoms, the most abundant molecule of which is C60Then is C70、C84Secondly a relatively low content of C76、C78、C82And the like. In addition, because the interior of the carbon cage of the fullerene is of a cavity structure, different atoms, ions or atom clusters can be embedded into the internal cavity of the fullerene, so that the fullerene is called as the embedded fullerene, such as La @ C60Indicating that La is embedded in C60In the cage structure of (1), @ represents at, and the embedded meaning is expressed visually.
The information disclosed in this background section is only for enhancement of understanding of the general background of the invention and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.
Disclosure of Invention
Technical problem
The technical problem to be solved by the invention is how to provide the application of the water-soluble fullerene structure in preparing the medicines for treating pancreatic cancer, pancreatitis and pancreatic cyst. Another technical problem to be solved by the present invention is to provide a pharmaceutical composition and a method for treating pancreatic cancer, pancreatitis and pancreatic cyst. The invention further aims to provide a health-care product for improving pancreatic cancer, pancreatitis and pancreatic cyst. The water-soluble fullerene structure of the effective component can be selectively enriched at the pancreatic part and treat pancreatic diseases such as pancreatitis and pancreatic cancer in a relatively short time.
Solution scheme
In order to solve the above technical problems, embodiments of the present invention provide the following technical solutions:
use of a water-soluble fullerene structure for the manufacture of a medicament for the treatment of pancreatic disease, said water-soluble fullerene structure comprising at least one of: a water-soluble modified hollow fullerene, a water-soluble modified endohedral metallofullerene, a combination of the water-soluble modified hollow fullerene and the water-soluble modified endohedral metallofullerene, pharmaceutically acceptable esters thereof or pharmaceutically acceptable salts thereof; the pancreatic diseases include pancreatic cancer, pancreatitis and pancreatic cysts.
The present invention also provides a method of treating pancreatic disease comprising administering to a subject having pancreatic disease an effective amount of at least one water-soluble fullerene structure selected from the group consisting of: a water-soluble modified hollow fullerene, a water-soluble modified endohedral metallofullerene, a composition of the water-soluble modified hollow fullerene and the water-soluble modified endohedral metallofullerene, pharmaceutically acceptable esters thereof or pharmaceutically acceptable salts thereof; the pancreatic diseases include pancreatic cancer, pancreatitis and pancreatic cysts.
The present invention also provides a pharmaceutical composition for treating pancreatic diseases, comprising at least one water-soluble fullerene structure selected from the group consisting of: a water-soluble modified hollow fullerene, a water-soluble modified endohedral metallofullerene, a combination of the water-soluble modified hollow fullerene and the water-soluble modified endohedral metallofullerene, pharmaceutically acceptable esters thereof or pharmaceutically acceptable salts thereof; the pharmaceutical composition further comprises at least one of a pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent, or a pharmaceutically acceptable excipient; the pancreatic diseases include pancreatic cancer, pancreatitis and pancreatic cysts.
The present invention also provides a health product for improving pancreatic diseases, which comprises at least one water-soluble fullerene structure selected from the group consisting of: a water-soluble modified hollow fullerene, a water-soluble modified endohedral metallofullerene, a composition of the water-soluble modified hollow fullerene and the water-soluble modified endohedral metallofullerene, pharmaceutically acceptable esters thereof or pharmaceutically acceptable salts thereof, the health care product further comprising at least one of a carrier suitable for a health care product, a diluent suitable for a health care product or an excipient suitable for a health care product; the pancreatic diseases include pancreatic cancer, pancreatitis and pancreatic cysts.
In another embodiment of the above use, method, pharmaceutical composition or health product, the water-soluble modified hollow fullerene comprises one or more selected from the group consisting of: (1) modifying the modified hollow fullerene with hydrophilic groups on the outer surface of the carbon cage of the hollow fullerene body; (2) the outer surface of a carbon cage of the hollow fullerene body is coated with hydrophilic biological micromolecules; (3) modified hollow fullerene formed by loading a hollow fullerene body with a biocompatible carrier material; (4) self-assembled to form the hollow fullerene of a water-soluble supermolecular system.
In another embodiment of the above use, method, pharmaceutical composition or health product, the water-soluble modified endohedral metallofullerene comprises one or more selected from the group consisting of: (1) the modified embedded metal fullerene with hydrophilic groups is modified on the outer surface of the carbon cage of the embedded metal fullerene body; (2) the outer surface of the carbon cage of the embedded metal fullerene body is coated with hydrophilic biological micromolecules to obtain modified embedded metal fullerene; (3) the embedded metal fullerene body is loaded by a carrier material with biocompatibility to form modified embedded metal fullerene; (4) the embedded metal fullerene of the water-soluble supermolecule system is formed by self-assembly.
In another embodiment of the above application, method, pharmaceutical composition or health product, the hollow fullerene body comprises one or more hollow fullerene compounds having a general formula C2mM is more than or equal to 20 and less than or equal to 60, m is more than or equal to 30 and less than or equal to 60, and m is more than or equal to 30 or 35 or 42.
In another embodiment of the above application, method, pharmaceutical composition or health product, the endohedral metallofullerene body comprises M @ C2n、M2@C2n、MA@C2n、M3N@C2n、M2C2@C2n、M2S@C2n、M2O@C2nAnd MxA3-xN@C2nOne or more ofWherein: m, A each represents a metal element and M, A each is selected from any of Sc, Y and lanthanide metal elements, 20. ltoreq. n.ltoreq.60, optionally 30. ltoreq. n.ltoreq.60, n being 41 or 30 or 35; x is more than or equal to 0 and less than or equal to 3. N represents nitrogen, C represents carbon, S represents sulfur, O represents oxygen, and the lanthanoid 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 use, method, pharmaceutical composition or nutraceutical, the hydrophilic group comprises one or more of hydroxyl, carboxyl, sulfhydryl, amino and hydrophilic amino acid residues.
In another embodiment of the above application, method, pharmaceutical composition or health product, the amino acid residue refers to an incomplete amino acid remaining after a part of an amino acid molecule is lost when the amino acid is modified in an empty fullerene body or an endohedral metallofullerene body, that is: an amino acid residue is a portion of an amino acid molecule that is an incomplete amino acid. Any portion of the molecule lacking amino acids is counted as an amino acid residue, such as: the amino acid loses hydrogen on the amino group to be bonded with the hollow fullerene body or the embedded metal fullerene body, and the amino acid loses hydrogen or hydroxyl on the carboxyl group to be bonded with the hollow fullerene body or the embedded metal fullerene body. Alternatively, the amino acid residue is a water-soluble amino acid residue, such as: at least one of an alanine residue, a glycine residue, a serine residue, an arginine residue, a lysine residue, and a aspartic acid residue.
In another embodiment of the above application, method, pharmaceutical composition or health product, the water-soluble modified hollow fullerene has a general formula of C2a(OH)b(ii) a A is more than or equal to 20 and less than or equal to 60, optionally more than or equal to 30 and less than or equal to 60, and further optionally a is 30 or 35 or 42; 0<b is less than or equal to 50, and 0 is optional<b is less than or equal to 30, b is less than or equal to 10 and less than or equal to 30, b is less than or equal to 20 and less than or equal to 30, and further optional b is 13, 20, 22, 24 or 26.
In another embodiment of the above application, method, pharmaceutical composition or health product, the water-soluble modified endohedral metallofullerene has the general formula metaloflulerene- (OH)c(ii) a methallovelene stands forBulk of the embedded metal fullerene, 0<c is less than or equal to 50, and optionally 0<c is less than or equal to 30, c is less than or equal to 10 and less than or equal to 30, c is less than or equal to 20 and less than or equal to 30, and further optional c is 13, 20, 22, 24 or 26.
In another embodiment of the above application, method, pharmaceutical composition or health product, the water-soluble modified endohedral metallofullerene has a general formula of M @ C2d(OH)eM is selected from rare earth metals, and optional rare earth metals are Gd or La; d is more than or equal to 20 and less than or equal to 60, further optional d is more than or equal to 30 and less than or equal to 60, and d is 41 or 30 or 35; 0<e is less than or equal to 50, and optional 0<e.ltoreq.30, e.ltoreq.20, e.ltoreq.30, and optionally e 13, 20, 22, 24 or 26.
In another embodiment of the above application, method, pharmaceutical composition or health product, the water-soluble modified hollow fullerene is C70(OH)24(ii) a The water-soluble modified metal-embedded fullerene is Gd @ C82(OH)26。
In the above general formula C2a(OH)b、metallofullerene-(OH)cAnd the general formula M @ C2d(OH)eRepresenting the attachment of a hydroxyl group to the hollow fullerene body or to the endohedral metallic fullerene body. In the general formula, b, c and e are all statistical average values calculated by detection.
In another embodiment of the above use, method, pharmaceutical composition or nutraceutical, the hydrophilic small biological molecule comprises at least one of an amino acid and a peptide chain.
In another embodiment of the above use, method, pharmaceutical composition or nutraceutical, said biocompatible carrier material comprises at least one of a liposome and a cell membrane carrier.
In another embodiment of the above application, method, pharmaceutical composition or health product, the water-soluble modified hollow fullerene is obtained by water-soluble modification of a hollow fullerene body, and the water-soluble modified endohedral metallofullerene is obtained by water-soluble modification of an endohedral metallofullerene body.
In another embodiment of the above use, method, pharmaceutical composition or health product, the method of water solubility modification is any one of the following methods:
(1) the method for modifying the hydroxyl on the surface comprises the following steps: mixing the hollow fullerene body and/or the embedded metal fullerene body, hydrogen peroxide and an alkali solution, reacting until the hollow fullerene body and/or the embedded metal fullerene body are completely dissolved, filtering the obtained reaction solution, washing the filtrate, collecting the precipitate, and dialyzing to obtain the water-soluble hydroxyl derivative corresponding to the body.
Optionally, the method for modifying the surface with hydroxyl groups comprises: (a) mixing 1-30 wt% hydrogen peroxide and 10-80 wt% sodium hydroxide aqueous solution and/or potassium hydroxide aqueous solution at a volume ratio of 1-10:1 to obtain a mixed solution, and adding 20-500mg hollow fullerene body and/or embedded metal fullerene body (such as C) into 10-200ml mixed solution60Solids or C70Solid or Gd @ C82Solid), stirring at 50-80 deg.C (optionally stirring for 4-24h at 1000r/min) until the solid is completely dissolved, filtering, and retaining the filtrate; (b) adding excessive ethanol with the concentration of 85% -100% into the filtrate for washing, collecting precipitate, and dissolving the precipitate in water to obtain a solution; (c) subjecting the solution obtained in step (b) to a dialysis treatment (optionally dialysis to a conductivity of the solution at room temperature of less than 1 μ s/cm). A further optional dialysis treatment is followed by a step of freeze-drying in order to obtain the corresponding solid.
Further optionally, the method for modifying the surface of the hydroxyl group comprises the following steps: (a) mixing hydrogen peroxide with the mass percentage of 20-30% and sodium hydroxide aqueous solution and/or potassium hydroxide aqueous solution with the mass percentage of 10-20% according to the volume ratio of 1-4:1 to obtain mixed solution, adding 20-150mg of hollow fullerene body and/or embedded metal fullerene body (such as C60 solid or C70 solid or Gd @ C82 solid) into every 10-20ml of the mixed solution, stirring at 50-60 ℃ until the solids are completely dissolved, filtering, and retaining filtrate; (b) adding excessive ethanol with the concentration of 85% -100% into the filtrate for washing, collecting precipitate, and dissolving the precipitate in water to obtain a solution; (c) dialyzing the solution obtained in step (b).
(2) The method for modifying the amino group on the surface comprises the following steps: and replacing the aqueous solution of sodium hydroxide and/or the aqueous solution of potassium hydroxide in the step with ammonia water.
(3) The physical coating method comprises the following steps: the hollow fullerene body and/or the embedded metal fullerene body is mixed with at least one of polyethylene glycol, polyvinylpyrrolidone and cyclodextrin, and the mixture is subjected to ball milling or ultrasonic treatment, so that a coated water-soluble fullerene structure corresponding to the body can be obtained, such as the hollow fullerene coated by polyethylene glycol and/or the embedded metal fullerene coated by polyethylene glycol, the hollow fullerene coated by polyvinylpyrrolidone and/or the embedded metal fullerene coated by polyvinylpyrrolidone.
In another embodiment, the above use, method or pharmaceutical composition for treating pancreatic disorders comprises (1) treating pancreatic gland swelling and necrosis, increased weight of the pancreas, increased water content of the pancreas resulting from pancreatitis; (2) reducing bloody ascites caused by pancreatitis; (3) reducing the content of serum amylase and lipase; (4) improving damage of other tissues and organs caused by pancreatic diseases; (5) inhibiting pancreatic tumor growth; (6) extending survival in patients with pancreatic cancer; (7) reduce the emaciation, weakness, abdominal pain or digestive tract problems caused by pancreatic diseases.
The drug or pharmaceutical composition of the above uses in another embodiment, the drug or pharmaceutical composition is in a pharmaceutically acceptable dosage form, which may be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions or sterile packaged powder injections. In the invention, the water-soluble fullerene structure of the active ingredient is prepared into a medicament or a medicinal composition, so that the active ingredient can be released quickly, slowly or slowly after being applied to a subject. During the preparation process, the active ingredient may be mixed with the carrier, or diluted with the carrier, or encapsulated in the carrier.
The medicament in the above application or the above pharmaceutical composition in another embodiment, when the medicament or the pharmaceutical composition is in a liquid form, the concentration of the water-soluble fullerene structure in the medicament or the pharmaceutical composition is 0.01-50mg/mL, optionally 0.01-5mg/mL, 0.01-10mg/mL, 5-10 mg/mL; when the drug or the pharmaceutical composition is in solid form, the concentration of the water-soluble fullerene structure in the drug or the pharmaceutical composition is 0.01-100mg/g, optionally 0.01-5mg/g, 5-10 mg/g.
The above-mentioned medicament or the above-mentioned pharmaceutical composition for use in another embodiment, preferably, the medicament or the pharmaceutical composition is an intraperitoneal injection.
In another embodiment, the concentration of the active ingredient in the intraperitoneal injection is 0.01-50mg/ml, optionally 0.5-5mg/ml and 0.5-10 mg/ml; the intraperitoneal injection comprises a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises at least one of water, physiological saline, PBS buffer solution and Tris-HCl solution; optionally, the concentration of the physiological saline is 0.85-0.90%; optionally, the concentration of PBS buffer solution is 0.01-0.1mol/L, and the component is specifically Na2HPO4、KH2PO4NaCl and KCl; optionally, the concentration of the Tris-HCl solution is 0.05 mol/L.
In another embodiment of the medicament for the above use or the above pharmaceutical composition, some examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, starch, resins, acacia gum, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup (water syrup), methyl cellulose, methyl and propyl parabens, talc, magnesium stearate and liquid paraffin. The formulations may additionally include lubricating agents, wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavoring agents.
In another embodiment of the above method, the subject is a mammal, such as: mouse, guinea pig, rat, dog, rabbit, monkey, human, and the like.
In another embodiment of the above method, the effective ingredient is administered in an amount of 1mg/kg/d to 100mg/kg/d, optionally 1 to 10mg/kg/d, 1 to 20mg/kg/d, 10 to 50mg/kg/d, and the total dose of the subject is converted to the body weight of the subject. The administration course can be 5-30 days, and can be short-term administration or long-term administration according to the disease condition. The administration mode of the effective components is preferably abdominal cavity administration, and compared with the abdominal cavity injection, other injection modes such as: when the medicine is administrated by intravenous injection and oral administration, the enrichment amount of pancreas is greatly reduced, and the curative effect on pancreatic diseases is poor.
The term "treating" as used herein includes its generally accepted meaning which includes preventing, inhibiting, and slowing, stopping or reversing the development of the symptoms or pathology desired. As such, the invention encompasses both therapeutic and prophylactic administration.
The term "effective amount" as used herein refers to an amount or dose of an active ingredient which, upon single or multiple administration to a patient, provides the desired effect in the patient diagnosed or treated. An effective amount can be determined by the attending diagnostician as one skilled in the art by known techniques and by observations made under similar circumstances. In determining the effective amount or dosage of the administered active ingredient, the attending diagnostician will consider a variety of factors including, but not limited to: mammalian species; volume, age, and general health; the particular disease involved; the degree or severity of involvement of the disease; the response of the individual patient; the specific compound administered; a mode of administration; the bioavailability properties of the administered formulation; the selected dosing regimen; use with drug therapy; and other related situations.
The term "hollow fullerene bulk" used in the present invention refers to hollow fullerene which is not water-soluble modified, i.e., a hollow fullerene raw material.
The term "bulk endohedral metallofullerene" used in the present invention refers to the endohedral metallofullerene without water-soluble modification, i.e., the endohedral metallofullerene raw material.
The disclosure of all ranges herein should be considered as disclosing all sub-ranges and all point values within the range. For example: the disclosure of 1-1000 should be considered to disclose the range of 1-200,200-300, etc., as well as 200, 300, 400, 500, 600, 700, 800, 900, and 100, etc.
Advantageous effects
The inventor creatively discovers that the water-soluble fullerene structure can be selectively and massively enriched at the pancreas when the water-soluble fullerene structure enters a living body in the form of intraperitoneal injection through intraperitoneal injection. The structure of the water-soluble fullerene enriched at the pancreas when the fullerene is injected into the abdominal cavity is 5 times that of the fullerene when the fullerene is injected into the vein and 100 times that of the fullerene when the fullerene is taken orally. And the intraperitoneal injection enters into organisms in an intraperitoneal injection mode, so that the water-soluble fullerene structure can directly interact with pancreas in an abdominal cavity, and the effect is obvious.
After entering the body, the water-soluble fullerene structure can treat pancreatic diseases, such as: inhibiting pancreatic tumor growth, treating pancreatic swelling caused by pancreatitis, and reducing serum amylase and lipase increase caused by pancreatitis.
The water-soluble fullerene structure as the effective component can be rapidly metabolized after entering the body, and has no toxicity to the organs in the body.
Other features and aspects of the present invention will become apparent from the following detailed description of exemplary embodiments, which proceeds with reference to the accompanying drawings.
Drawings
One or more embodiments are illustrated by the corresponding figures in the drawings, which are not meant to be limiting.
FIG. 1A shows C prepared in example 170Thermogravimetric analysis and differential quotient thermogravimetric curves of corresponding hydroxylated derivatives; FIG. 1B is Gd @ C prepared in example 182Thermogravimetric analysis and differential quotient thermogravimetric curves of the corresponding hydroxylated derivatives.
FIG. 2 is Gd @ C obtained in example 182(OH)nParticle size distribution of the material in aqueous solution.
FIG. 3 is Gd @ C of example 282(OH)nDistribution of the material in different tissues and organs 12h after entering the body of the mouse.
FIG. 4 is a plot of pancreatic tumor growth in two groups of tumor-bearing mice from example 3. The squares represent physiologySaline control group, circle represents Gd @ C82(OH)nAnd (4) treatment groups.
FIG. 5 is a graph showing pancreatic coefficients of the groups of mice in example 4.
FIG. 6 shows the intraperitoneal injection of Gd @ C82(OH)nAfter 24h, its metabolic distribution in the organism.
FIG. 7 is an intravenous Gd @ C injection82(OH)nAfter 24h, its metabolic distribution in the organism.
FIG. 8 is oral administration of Gd @ C82(OH)nAfter 24h, its metabolic distribution in the organism.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. Throughout the specification and claims, unless explicitly stated otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or component but not the exclusion of any other element or component.
The word "exemplary" is used exclusively in this application to mean "serving as an example, embodiment, or illustration. Any embodiment described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments.
Furthermore, in the following detailed description, numerous specific details are set forth in order to provide a better understanding of the present invention. It will be understood by those skilled in the art that the present invention may be practiced without some of these specific details. In some instances, methods, means, elements well known to those skilled in the art have not been described in detail so as not to obscure the present invention.
Materials, reagents, etc. used in the examples described below, e.g. WuteIt is well described and can be obtained from commercial sources. The experimental methods used in the following examples are all conventional experimental methods unless otherwise specified. Gd @ C as a starting material for the following examples82The solid powder was purchased from Xiamen New materials science and technology Inc., molecular weight 1141, purity 99.1%. The starting material C70 solid powder used in the following examples was purchased from Xiamen New materials science and technology, Inc., molecular weight 840, purity 99%.
Example 1 Water-soluble hollow Fullerene C70(OH)nWater soluble embedded metal fullerene Gd @ C82(OH)nPreparation and particle size testing of
Weighing 100mgC70Mixing the solid powder, 7ml of 30% hydrogen peroxide and 3ml of 10% sodium hydroxide aqueous solution, and reacting at 50 ℃ to obtain the product C70Filtering after the solid powder is completely dissolved; the resulting filtrate was washed with excess ethanol to remove excess H2O2And NaOH, centrifuging (rotation speed: 10000r/min, time: 4min), removing upper colorless liquid, dissolving the collected precipitate in ultrapure water to obtain a clear solution, dialyzing the clear solution in ultrapure water by using a M.W. ═ 3500 dialysis bag until the conductivity of the ultrapure water is less than 1 mu s/cm, and freeze-drying to obtain the hollow fullerene hydroxylated derivative C70(OH)nAnd (3) a solid.
Weighing 100mgGd @ C82Mixing the solid powder, 7ml of 30% hydrogen peroxide and 3ml of 10% sodium hydroxide aqueous solution, and reacting at 50 ℃ until Gd @ C82Filtering after the solid powder is completely dissolved; the resulting filtrate was washed with excess ethanol to remove excess H2O2And NaOH, removing upper colorless liquid after centrifugation (rotation speed: 10000r/min, time: 4min), dissolving the collected precipitate in ultrapure water to obtain a clear solution, dialyzing the clear solution in ultrapure water by using a M.W. ═ 3500 dialysis bag until the conductivity of the ultrapure water is less than 1 mu s/cm, and freeze-drying to obtain the embedded metal fullerene hydroxylated derivative Gd @ C82(OH)n。
For C prepared above70(OH)nGo on yuanAnd (4) analyzing element (Flash EA 1112) and analyzing the number of the connected hydroxyl groups by combining thermogravimetric and differential quotient thermogravimetric results. In the results of the elemental analysis, C70(OH)nIn the formula, the content of C is 37.85%, the content of H is 1.51%, and the content of N is<0.3 percent. From the thermogravimetric analysis of FIG. 1A, C70(OH)nThe solid powder contains 3.7 percent of water, and the ratio of the H content to the C content in element analysis can be used for calculating that 24 hydroxyl groups are modified on the surface of the carbon cage. So C70(OH)nHas an average structural formula of C70(OH)24。
For Gd @ C obtained by the preparation82(OH)nAnd (4) performing element analysis, and analyzing the number of connected hydroxyl groups by combining thermogravimetric and differential quotient thermogravimetric results. In the results of elemental analysis, the Gd @ C82(OH)nIn the formula, the content of C is 36.95%, the content of H is 2.36% and the content of N is 0%. From the thermogravimetric analysis of FIG. 1B, Gd @ C82(OH)nThe solid powder contains 12.6 percent of water, and 26 hydroxyl groups modified on the surface of the carbon cage can be calculated by combining the ratio of the H content to the C content in the element analysis. Thus Gd @ C82(OH)nHas an average structural formula of Gd @ C82(OH)26. Particle size of the water-soluble derivative was determined by Dynamic Light Scattering (DLS), as shown in FIG. 2, Gd @ C82(OH)nThe particle size of the material is about 145.2nm, and the material can be injected into organisms.
Example 2 Water-soluble Metal-Embedded Fullerene Gd @ C82(OH)nIn vivo metabolism of
1mM of water-soluble metal-embedded fullerene Gd @ C82(OH)nThe injection was made into C57 mice via the abdominal cavity at a dose of 10 ml/kg. Dissecting and taking materials after 12h, weighing heart, liver, spleen, lung, kidney, pancreas, muscle, subcutaneous fat, brown fat, stomach and small intestine, digesting with 65% nitric acid at 100 deg.C for 24h, diluting the digestion solution by 50 times, and measuring gadolinium ion concentration in the digestion solution by ICP test, the result is shown in figure 3, and water-soluble embedded metal fullerene Gd @ C can be obtained82(OH)nThe most abundant in pancreas shows that the water-soluble fullerene structure of the invention can be abundantly enriched in pancreas, and provides treatment for pancreatic diseasesAnd (4) a foundation.
Example 3 Water-soluble Metal-Embedded Fullerene Gd @ C82(OH)nApplication in treating pancreatic cancer
(1) Establishment of pancreatic cancer model
Pancreatic cancer cell JF305 is cultured to a certain number, digested into cell suspension, washed for 3 times by PBS, adjusted to cell suspension with cell density of 1 x 10^ 7/ml, 150 microliters of cell suspension is inoculated to the subcutaneous part of balb/c nude mice, and fed until tumors grow to the diameter of about 4-5 mm.
(2) Water-soluble metal-embedded fullerene Gd @ C82(OH)nExperiment for treating pancreatic cancer
Randomly dividing balb/C tumor-bearing mice obtained in the step (1) into a normal saline control group and Gd @ C82(OH)nTreatment groups, 6 per group. Saline control group and Gd @ C82(OH)nThe treatment groups all adopt an abdominal cavity administration mode. Gd @ C82(OH)nGd @ C administration in treatment groups82(OH)nIntraperitoneal injection (namely Gd @ C prepared in example 1)82(OH)nGd @ C formed by dissolving in 0.9% physiological saline82(OH)nIntraperitoneal injection solution with concentration of 10 mg/ml), wherein Gd @ C82(OH)nThe administration dose of (2) was 10mg/kg/d, the saline control group was administered with the same volume of saline as the treatment group as a control, both groups were continuously administered for 2 weeks, and the tumor size was observed and measured with a vernier caliper every other day. FIG. 4 is a plot of pancreatic tumor growth in two groups of tumor-bearing mice. Gd @ C is obtained from FIG. 482(OH)nThe treated pancreatic tumor grows obviously slowly, and the tumor size is obviously reduced, which shows that the water-soluble fullerene structure can inhibit the growth of the pancreatic tumor and can effectively treat pancreatic cancer.
Example 4 Water-soluble hollow Fullerene C70(OH)nApplication in treating pancreatitis
(1) Establishment of pancreatitis model and treatment scheme
Female C57BL/6 mice 18 were evenly divided into three groups: (1) normal mouse control group; (2) a pancreatitis model group; (3) and (4) treatment groups. Model of pancreatitis inMice in the groups and the treatment groups need to establish a mouse pancreatitis model in a mode of combining ranophanin with lipopolysaccharide modeling. The preparation method comprises the steps of carrying out intraperitoneal injection on 50 microgram/kg of ranulin to a molded animal after fasting for 18 hours before an experiment, continuously injecting for six times at an interval of 1 hour each time, and injecting 10mg/kg of lipopolysaccharide to the abdominal cavity after each injection of the ranulin. Treatment group injected with C1, 5, 9, 12h after the first injection of ranolanin70(OH)nIntraperitoneal injection (i.e. C prepared in example 1)70(OH)nC formed by dissolving in 0.9% physiological saline70(OH)nIntraperitoneal injection solution with concentration of 10 mg/ml), wherein C is70(OH)nThe administration dose of (a) is 10mg/kg/d, i.e. the water-soluble hollow fullerene C is injected both during and after the modeling70(OH)nNormal mouse control group was injected with physiological saline according to the same procedure. The materials are taken 18 hours after treatment, the mice are weighed and then killed, whole blood is taken, the whole blood is stood still and then centrifuged, 2500 rpm is carried out, 15 minutes are carried out, the contents of serum amylase and lipase are measured, and the weight of pancreas is weighed.
(2) Therapeutic results
As shown in fig. 5, the pancreas coefficient (pancreas coefficient ═ pancreas weight/mouse weight) increased in the model group, C70(OH)nThe pancreas coefficient of the mice is obviously reduced and tends to be normal after treatment, which shows that the water-soluble fullerene structure can obviously improve pancreas swelling and mass increase caused by pancreatitis.
In addition, the levels of serum amylase and lipase were also examined. Serum amylase is the main type of amylase in serum, belongs to glycoside chain hydrolase, is mainly derived from pancreas and the like, and is mainly used for diagnosing acute pancreatitis in activity measurement. The lipase is an enzyme for hydrolyzing long-chain fatty glyceride, and the lipase in serum mainly comes from pancreas, and the increase of the lipase is parallel to amylase when in pancreatitis, so the lipase can be used for diagnosing pancreatitis. Serum amylase and lipase, both significantly elevated in the model group, as shown in Table 1, via water-soluble hollow fullerene C70(OH)nThe obvious reduction after treatment also indicates that the water-soluble fullerene can treat pancreatitis.
Table 1 serum amylase and lipase levels for each group of mice.
Example 5 Water-soluble Embedded Metal Fullerene Gd @ C under different injection modes82(OH)nIn vivo metabolism of
1mM of the metallofullerene Gd @ C prepared according to the method in example 1 was taken82(OH)nAnd the medicine is injected into C57 mice by intravenous injection, oral administration and intraperitoneal injection respectively, and the dosage is 10 ml/kg. Materials were dissected 24h later, heart, liver, spleen, lung, kidney, pancreas were weighed and digested with 65% nitric acid at 100 ℃ for 24h, diluted 50-fold and then determined for gadolinium ion concentration by ICP test.
As shown in FIG. 6, water-soluble endohedral metallofullerene Gd @ C was administered by means of intraperitoneal injection82(OH)nThereafter, it was most enriched in the pancreas, where it reached 4.5. mu.g/g. As shown in FIG. 7, water-soluble endohedral metallofullerene Gd @ C was injected intravenously82(OH)nLater, it was mainly enriched in the liver, and very little in the pancreas, less than 1.0. mu.g/g. As shown in FIG. 8, after intragastric administration, the water-soluble endohedral metallofullerene Gd @ C82(OH)nEnrichment was less, even though it was most abundant in the pancreas, but only about 0.035. mu.g/g. This demonstrates that the water-soluble fullerene structure of the present invention can be greatly enriched at the pancreas by intraperitoneal injection, and is five times as large as that of the vein and 100 times as large as that of oral administration.
The invention uses water-soluble hollow fullerene C70(OH)nAnd water-soluble embedded metal fullerene Gd @ C82(OH)nFor example, it was found that such materials can be selectively enriched at the pancreas and can be effective in treating pancreas-related diseases such as pancreatic cancer, pancreatitis, pancreatic cysts, and the like.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (11)
1. The application of a water-soluble fullerene structure in preparing an intraperitoneal injection for treating pancreatic diseases is characterized in that the water-soluble fullerene structure is water-soluble modified hollow fullerene; the water-soluble modified hollow fullerene is C70(OH)24(ii) a The pancreatic disease is pancreatitis.
2. The use of claim 1, wherein the intraperitoneal injection solution further comprises at least one of a pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent, or a pharmaceutically acceptable excipient.
3. The use of claim 2, wherein the concentration of the active ingredient of the intraperitoneal injection solution is 0.01mg/ml to 50 mg/ml.
4. The use of claim 3, wherein the concentration of the active ingredient of the intraperitoneal injection solution is optionally 0.01mg/ml to 10 mg/ml.
5. The use of claim 2, wherein the pharmaceutically acceptable carrier comprises at least one of water, physiological saline, PBS buffer, and Tris-HCl solution.
6. The use according to claim 5, wherein the physiological saline is in a concentration of 0.85-0.90%.
7. The use of claim 5, wherein the concentration of the PBS buffer is 0.01-0.1 mol/L.
8. The use according to claim 5, wherein the concentration of the Tris-HCl solution is 0.05 mol/L.
9. Use according to any one of claims 1 to 8, wherein the hollow fullerene body is C70。
10. The use according to any one of claims 1-8, wherein the water-soluble modified hollow fullerene is obtained by water-soluble modification of a bulk hollow fullerene by: weighing 100mgC70Mixing the solid powder, 7ml of 30% hydrogen peroxide and 3ml of 10% sodium hydroxide aqueous solution, and reacting at 50 ℃ to obtain the product C70Filtering after the solid powder is completely dissolved; the resulting filtrate was washed with excess ethanol to remove excess H2O2And NaOH and centrifugation, speed: 10000r/min, time: removing the upper layer colorless liquid after 4min, dissolving the collected precipitate in ultrapure water to obtain a clear solution, dialyzing the clear solution in ultrapure water by using a M.W. ═ 3500 dialysis bag, dialyzing at room temperature until the conductivity of the ultrapure water is less than 1 mus/cm, and freeze-drying.
11. The use of any one of claims 1-8, wherein treating pancreatic disease comprises (1) treating pancreatic gland swelling and necrosis, increased pancreatic weight, increased pancreatic water content resulting from pancreatitis; (2) reduce the content of serum amylase and lipase.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104127873A (en) * | 2014-07-29 | 2014-11-05 | 中国科学院化学研究所 | Nano material and external radiation source-containing tumor treatment medicine for realizing tumor vascular occlusion |
| CN104127872A (en) * | 2014-07-29 | 2014-11-05 | 中国科学院化学研究所 | Application of metal fullerene monocrystal nanoparticles in preparation of specific tumor vascular disrupting agent |
| CN105903021A (en) * | 2016-03-25 | 2016-08-31 | 北京福纳康生物技术有限公司 | Fullerene nano-material for preventing and/or treating aplastic anemia and use thereof |
| CN106267238A (en) * | 2015-05-14 | 2017-01-04 | 中国科学院化学研究所 | A kind of metal fullerene complex and preparation method and application |
| CN106491644A (en) * | 2016-11-25 | 2017-03-15 | 北京福纳康生物技术有限公司 | A kind of fullerene soft capsule and preparation method |
| CN106491547A (en) * | 2016-11-25 | 2017-03-15 | 北京福纳康生物技术有限公司 | A kind of endohedral tablet and preparation method |
| CN106620727A (en) * | 2016-10-08 | 2017-05-10 | 北京福纳康生物技术有限公司 | Amino acid modified metallofullerene water-soluble nanoparticles as well as preparation method and application thereof |
| CN106619715A (en) * | 2016-10-08 | 2017-05-10 | 北京福纳康生物技术有限公司 | Application of amino-acid-modified metallofullerene water-soluble nanoparticles in preparation of tumor vascular disrupting agents |
| CN107137423A (en) * | 2016-10-08 | 2017-09-08 | 北京福纳康生物技术有限公司 | A kind of water-soluble fullerene nano material and preparation method and application |
-
2017
- 2017-12-19 CN CN201711376934.5A patent/CN109925322B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104127873A (en) * | 2014-07-29 | 2014-11-05 | 中国科学院化学研究所 | Nano material and external radiation source-containing tumor treatment medicine for realizing tumor vascular occlusion |
| CN104127872A (en) * | 2014-07-29 | 2014-11-05 | 中国科学院化学研究所 | Application of metal fullerene monocrystal nanoparticles in preparation of specific tumor vascular disrupting agent |
| CN106267238A (en) * | 2015-05-14 | 2017-01-04 | 中国科学院化学研究所 | A kind of metal fullerene complex and preparation method and application |
| CN105903021A (en) * | 2016-03-25 | 2016-08-31 | 北京福纳康生物技术有限公司 | Fullerene nano-material for preventing and/or treating aplastic anemia and use thereof |
| CN106620727A (en) * | 2016-10-08 | 2017-05-10 | 北京福纳康生物技术有限公司 | Amino acid modified metallofullerene water-soluble nanoparticles as well as preparation method and application thereof |
| CN106619715A (en) * | 2016-10-08 | 2017-05-10 | 北京福纳康生物技术有限公司 | Application of amino-acid-modified metallofullerene water-soluble nanoparticles in preparation of tumor vascular disrupting agents |
| CN107137423A (en) * | 2016-10-08 | 2017-09-08 | 北京福纳康生物技术有限公司 | A kind of water-soluble fullerene nano material and preparation method and application |
| CN106491644A (en) * | 2016-11-25 | 2017-03-15 | 北京福纳康生物技术有限公司 | A kind of fullerene soft capsule and preparation method |
| CN106491547A (en) * | 2016-11-25 | 2017-03-15 | 北京福纳康生物技术有限公司 | A kind of endohedral tablet and preparation method |
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