CN115444943A - Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis - Google Patents

Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis Download PDF

Info

Publication number
CN115444943A
CN115444943A CN202211148299.6A CN202211148299A CN115444943A CN 115444943 A CN115444943 A CN 115444943A CN 202211148299 A CN202211148299 A CN 202211148299A CN 115444943 A CN115444943 A CN 115444943A
Authority
CN
China
Prior art keywords
cysteine
ligand
copper
arginine
liver
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202211148299.6A
Other languages
Chinese (zh)
Inventor
孙涛垒
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Wuhan Guanghang Scientific Research Co ltd
Original Assignee
Wuhan Guanghang Scientific Research Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Wuhan Guanghang Scientific Research Co ltd filed Critical Wuhan Guanghang Scientific Research Co ltd
Publication of CN115444943A publication Critical patent/CN115444943A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6941—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a granulate or an agglomerate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/34—Copper; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/542—Carboxylic acids, e.g. a fatty acid or an amino acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Inorganic Chemistry (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The present invention provides ligand-bound copper clusters (CuCs), formulations containing ligand-bound copper clusters, and their use in treating patients with liver cirrhosis.

Description

Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis
The invention is a divisional application of invention patents based on application numbers of 2020113065781, application date of 11/19/2020, entitled "compatibility of ligand-bound copper clusters, ligand-bound copper cluster-containing copper clusters and application thereof in treating liver cirrhosis".
Technical Field
The present invention relates generally to the field of liver cirrhosis treatment technology, and more particularly to the formulation of ligand-bound copper clusters (CuCs), ligand-bound copper cluster-containing formulations, and their use in the treatment of liver cirrhosis.
Background
The liver is the largest solid organ of the human body and has many important functions, including: manufacturing blood proteins that aid in clotting, oxygen delivery, and the immune system; and storing excess nutrients and returning a portion of the nutrients to the blood; making bile to aid in the digestion of food; help the body store sugar (glucose) in the form of glycogen; eliminating harmful substances in vivo, including drugs and alcohol; breakdown saturated fats and produce cholesterol.
Cirrhosis is a slowly progressive disease that develops over many years due to long-term, persistent liver damage. As cirrhosis progresses, healthy liver tissue is gradually destroyed and replaced by scar tissue. These scar tissues can prevent blood flow through the liver and slow the liver's ability to process nutrients, hormones, drugs and natural toxins. It also reduces the production of proteins and other substances produced by the liver. Cirrhosis may ultimately lead to liver failure and/or liver cancer that may require liver transplantation.
In the early stage of cirrhosis, the liver compensation function is strong, and no obvious symptoms exist. The late stage symptoms comprise complications such as liver function damage, portal hypertension, upper gastrointestinal hemorrhage, hepatic encephalopathy, secondary infection, spleen hyperfunction, ascites, canceration, etc. The liver gradually becomes deformed and hardened, and cirrhosis progresses. Histopathologically, cirrhosis manifests itself as extensive hepatocyte necrosis, nodular regeneration of residual hepatocytes, connective tissue proliferation and fibroseptal formation, leading to destruction of hepatic lobular structures and formation of false lobules.
Cirrhosis has different causes. Some patients with cirrhosis have liver damage due to a variety of causes. Common causes of cirrhosis include long-term alcohol abuse, chronic hepatitis b and c infection, fatty liver disease, toxic metals, genetic disorders, nutritional disorders, industrial poisons, pharmaceuticals, blood circulation disorders, metabolic disorders, cholestasis, schistosomiasis, and the like.
Cirrhosis can be diagnosed by a number of tests/techniques. For example, if liver enzymes including alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) and bilirubin levels are elevated and blood protein levels are reduced, a blood test may indicate cirrhosis.
Currently, although treatment can delay the progression of cirrhosis by eliminating the cause of cirrhosis, there is no specific treatment for cirrhosis.
Disclosure of Invention
The present invention provides for treating a subject having cirrhosis with ligand-bound copper clusters (CuCs) comprising a copper core, and a ligand that binds to the copper core to form ligand-bound copper clusters.
In some embodiments for this therapeutic application, the diameter of the copper core is 0.5-5nm. In some embodiments, the copper core has a diameter of 0.5 to 3nm.
In some embodiments of this therapeutic application, the ligand is one selected from the group consisting of thymine, thymine Modified Hyaluronic Acid (TMHA), L-cysteine and its derivatives, D-cysteine and its derivatives, cysteine-containing oligopeptides and derivatives, and other thiol-containing compounds.
In some embodiments of this therapeutic application, L-cysteine and its derivatives are selected from L-cysteine, N-isobutyryl-L-cysteine (L-NIBC), and N-acetyl-L-cysteine (L-NAC), and D-cysteine and its derivatives are selected from D-cysteine, N-isobutyryl-D-cysteine (D-NIBC), and N-acetyl-D-cysteine (D-NAC).
In some embodiments of the therapeutic application, the cysteine-containing oligopeptide and derivative thereof is a cysteine-containing dipeptide, a cysteine-containing tripeptide, or a cysteine-containing tetrapeptide.
In some embodiments of this therapeutic application, the cysteine-containing dipeptide is selected from the group consisting of L (D) -cysteine-L (D) -arginine dipeptide (CR), L (D) -arginine-L (D) -cysteine dipeptide (RC), L (D) -histidine-L (D) -cysteine dipeptide (HC), and L (D) -cysteine-L (D) -histidine dipeptide (CH).
In some embodiments for this therapeutic use, the cysteine-containing tripeptide is selected from the group consisting of glycine-L (D) -cysteine-L (D) -arginine tripeptide (GCR), L (D) -proline-L (D) -cysteine-L (D) -arginine tripeptide (PCR), L (D) -lysine-L (D) -cysteine-L (D) -proline tripeptide (KCP), and L (D) -Glutathione (GSH).
In some embodiments of this therapeutic use, the cysteine-containing tetrapeptide is selected from the group consisting of glycine-L (D) -serine-L (D) -cysteine-L (D) -arginine tetrapeptide (GSCR) and glycine-L (D) -cysteine-L (D) -serine-L (D) -arginine tetrapeptide (GCSR).
In some embodiments of this therapeutic application, the other thiol-containing compound is selected from the group consisting of 1- [ (2S) -2-methyl-3-thiol-1-oxopropyl ] -L (D) -proline, thioglycolic acid, mercaptoethanol, thiophenol, D-3-mercaptovaline, N- (2-mercaptopropionyl) -glycine, and dodecyl mercaptan.
The invention also applies ligand-bound copper clusters (CuC) comprising a copper core, and a ligand that binds to the copper core to form ligand-bound copper clusters for the preparation of a medicament for treating cirrhosis of the liver in a subject.
In some embodiments for use in the manufacture of a medicament, the copper core has a diameter of 0.5 to 5nm. In some embodiments, the copper core has a diameter of 0.5 to 3nm.
In some embodiments for use in the manufacture of a medicament, the ligand is one selected from thymine, thymine Modified Hyaluronic Acid (TMHA), L-cysteine and its derivatives, D-cysteine and its derivatives, cysteine-containing oligopeptides and their derivatives, and other thiol-containing compounds.
In some embodiments for the use of the medicament, L-cysteine and derivatives thereof are selected from L-cysteine, N-isobutyryl-L-cysteine (L-NIBC), and N-acetyl-L-cysteine (L-NAC), and D-cysteine and derivatives thereof are selected from D-cysteine, N-isobutyryl-D-cysteine (D-NIBC), and N-acetyl-D-cysteine (D-NAC).
In some embodiments for the pharmaceutical preparation use, the cysteine-containing oligopeptide and derivative thereof is a cysteine-containing dipeptide, a cysteine-containing tripeptide, or a cysteine-containing tetrapeptide.
In some embodiments of the pharmaceutical preparation use, the cysteine-containing dipeptide is selected from the group consisting of L (D) -cysteine-L (D) -arginine dipeptide (CR), L (D) -arginine-L (D) -cysteine dipeptide (RC), L (D) -histidine-L (D) -cysteine dipeptide (HC), and L (D) -cysteine-L (D) -histidine dipeptide (CH).
In some embodiments for use in the manufacture of a medicament, the cysteine-containing tripeptide is selected from the group consisting of glycine-L (D) -cysteine-L (D) -arginine tripeptide (GCR), L (D) -proline-L (D) -cysteine-L (D) -arginine tripeptide (PCR), L (D) -lysine-L (D) -cysteine-L (D) -proline tripeptide (KCP), and L-Glutathione (GSH).
In some embodiments of the pharmaceutical use of the invention, the cysteine-containing tetrapeptide is selected from the group consisting of glycine-L (D) -serine-L (D) -cysteine-L (D) -arginine tetrapeptide (GSCR) and glycine-L (D) -cysteine- (D) L-serine-L (D) -arginine tetrapeptide (GCSR).
In some embodiments for use in the manufacture of a medicament, the other thiol-containing compound is selected from the group consisting of 1- [ (2S) -2-methyl-3-thiol-1-oxopropyl ] -L (D) -proline, thioglycolic acid, mercaptoethanol, thiophenol, D-3-mercaptovaline, N- (2-mercaptopropionyl) -glycine, and dodecyl mercaptan.
The objects and advantages of the present invention will be apparent from the detailed description of the preferred embodiments taken in conjunction with the accompanying drawings.
Drawings
Preferred embodiments of the present invention will now be described with reference to the drawings, wherein like reference numerals represent like elements.
FIG. 1 shows characteristic data of L-glutathione-bound copper clusters (GSH-CuCs). (A) Typical Transmission Electron Microscope (TEM) images of GSH-CuCs. (B) GSH-CuCs size distribution calculated from TEM images. (C) X-ray photoelectron Spectroscopy (XPS) of 2p3/2 and 2p1/2 electrons of copper (0) in GSH-CuCs. (D) Fourier transform infrared spectroscopy (FT-IR) comparison of GSH-CuCs (top) and GSH (bottom). (E) Fluorescence excitation (left) and emission spectra (right) of GSH-CuCs.
FIG. 2 shows the effect of different doses of Cu-1 and Cu-2 administration on serum (A) ALT, (B) AST, (C) TBIL, (D) MAO and (E) ALB levels in cirrhosis model mice, with sorafenib treatment as the positive control group.
Fig. 3 shows HE staining pathology detection results: (a) a blank control group; (B) model control group; (C) a positive control group; (D) Cu-1 copper cluster low dose administration group; (E) Cu-1 copper cluster high dose administration group.
Detailed Description
The present invention may be understood more readily by reference to the following detailed description of certain embodiments of the invention.
Throughout this application, the disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.
The ligand-bound copper cluster comprises a copper core consisting of 2 to several hundred copper atoms, and a ligand; the ligand is bound to the copper core as a component of its molecule, forming a ligand-bound copper cluster that is stable in solution. Due to the low copper atom contrast, it is difficult to give a very accurate copper nucleus size by transmission electron microscopy, generally speaking, the copper nucleus size is 0.5-5nm by transmission electron microscopy.
The present invention provides copper clusters (CuCs) bound by one or more ligands, wherein the copper core is bound to the one or more ligands. The binding of the ligand to the copper core means that the ligand forms a complex stable in solution together with the copper core through a covalent bond, a hydrogen bond, an electrostatic force, a hydrophobic force or a van der waals force, or the like. In some embodiments, the diameter of the copper core is in the range of 0.5-5nm, preferably 0.5-3nm, more preferably 0.5-2.5 nm.
In some embodiments, ligands include, but are not limited to, thymine Modified Hyaluronic Acid (TMHA), L-cysteine, D-cysteine, and other cysteine derivatives, such as N-isobutyryl-L-cysteine (L-NIBC), N-isobutyryl-D-cysteine (D-NIBC), N-acetyl-L-cysteine, N-acetyl-D-cysteine, and the like; cysteine-containing oligopeptides and derivatives thereof, including but not limited to cysteine-containing dipeptides, tripeptides, tetrapeptides, and other peptides, such as: l (D) -cysteine-L (D) -arginine dipeptide (CR), L (D) -arginine-L (D) -cysteine dipeptide (RC), L (D) -cysteine L (D) -histidine (CH), glycine-L (D) -cysteine-L (D) -arginine tripeptide (GCR), L (D) -proline-L (D) -cysteine-L (D) -arginine tripeptide (PCR), L (D) -Glutathione (GSH), glycine-L (D) -serine-L (D) -cysteine-L (D) -arginine tetrapeptide (GSCR), glycine-L (D) -cysteine-L (D) -serine-L (D) -arginine tetrapeptide (GCSR), etc.; and other mercapto group-containing compounds such as one or more of 1- [ (2S) -2-methyl-3-mercapto-1-oxopropyl ] -L (D) -proline, thioglycolic acid, mercaptoethanol, thiophenol, D-3-mercaptovaline, dodecanethiol, and the like.
Copper clusters bound by different ligands were synthesized using methods described in the relevant literature (Deng 2018.
The present invention provides a pharmaceutical composition for treating cirrhosis of the liver in a subject. In some embodiments, the pharmaceutical composition comprises a ligand-bound copper cluster as disclosed above and a pharmaceutically acceptable excipient. In some embodiments, the excipient is a phosphate buffered solution or physiological saline. In some embodiments, the subject is a human. In some embodiments, the subject is a pet animal, e.g., a dog.
The present invention provides the use of the above disclosed ligand-bound copper clusters for the manufacture of a medicament for the treatment of liver cirrhosis in a subject.
The present invention provides the use of the above disclosed ligand-bound copper clusters for treating liver cirrhosis in a subject or a method of treating liver cirrhosis in a subject using the above disclosed ligand-bound copper clusters. In some embodiments, the method of treatment comprises administering to the subject a pharmaceutically effective amount of ligand-bound copper clusters. The pharmaceutically effective amount can be determined by routine in vivo studies.
The following examples are provided for the sole purpose of illustrating the principles of the present invention; they are in no way intended to limit or otherwise narrow the scope of the present invention.
EXAMPLE one Synthesis of TMHA-bound copper clusters (TMHA-CuCs)
10mL of a TMHA (DS 10.5%) solution (0.1 mM, pH 7.0) was gradually heated to 37 ℃ to dissolve the TMHA. 2mL of a copper sulfate (20mM, pH 7.0) solution was added dropwise thereto, and the reaction was further carried out at 37 ℃ in the dark for 20 minutes. Under UV light (365 nm), a bright orange-red emission was clearly visible, indicating successful synthesis of TMHA-bound copper clusters. Finally, the resulting solution was stored in the dark at 4 ℃ for future use. The diameter of the copper core of the spherical TMHA-CuCs is 1.64 +/-0.48 nm.
EXAMPLE two Synthesis and characterization of ligand-bound copper clusters with different ligands
2.1 Synthesis of L-Glutathione (GSH) -bound copper clusters
To 50mL of water was added 500mg of Glutathione (GSH), and 20mL of a 5mM Cu (NO 3) 2 solution was added with slow stirring. The solution quickly produced a white suspension. The mixture was slowly heated to 50-60 ℃ and heating continued for 20min, and 1M NaOH solution was added dropwise until the solution became pale yellow, clear and transparent. The product was cooled to room temperature and precipitated by adding several volumes of ethanol, repeated three times.
2.2 Synthesis of L-cysteine-bound copper clusters
50mL of CuCl2 at a concentration of 10mM was slowly added dropwise to a freshly prepared solution of L-cysteine (50mL, 10mM) with vigorous stirring. After about 30 minutes, 0.5mL of NaOH (1M) was slowly added drop-wise to the above solution. The reaction was continued for 2h. The product was centrifuged at 8000rpm for 20min and the supernatant was stored at 4 ℃ in the dark.
2.3 Synthesis of PEG-conjugated copper clusters
2.5g of PEG-SH (molecular weight 2000 or 5000) were dissolved in 100ml of ultrapure water at room temperature, and 4ml of a 0.5M Cu (NO 3) 2 solution was added dropwise with vigorous stirring. Stirring the mixed solution at room temperature for a period of time until the color is faded and gradually becomes milky white; the hydrogel was gradually heated to 80 ℃ and held for 15 minutes. A 3M NaOH solution was added dropwise until the solution became clear and transparent. The product was centrifuged at 8000rpm for 20min and the final product was freeze dried in a freeze dryer to give a solid sample.
2.4 Synthesis of ligand-bound copper clusters with other ligands
Ligand-bound copper clusters with other ligands can also be synthesized using the methods described above; the specific synthetic method needs to be slightly modified in some solvents and operation; other ligands include thymine, L (D) -cysteine and other cysteine derivatives, such as N-isobutyryl-L-cysteine (L-NIBC), N-isobutyryl-D-cysteine (D-NIBC), N-acetyl-L-cysteine and N-acetyl-D-cysteine, cysteine-containing oligopeptides and derivatives thereof, including but not limited to dipeptides, tripeptides, tetrapeptides and other cysteine-containing peptides, such as L (D) -cysteine-L (D) -arginine dipeptide (CR), L (D) -arginine-L (D) -cysteine dipeptide (RC), L (D) -cysteine L (D) -histidine (CH), glycine-L (D) -cysteine-L (D) -arginine tripeptide (GCR), L (D) -proline-L (D) -cysteine-L (D) -arginine tripeptide (PCR), L (D) -Glutathione (GSH), glycine-L (D) -serine-L (D) -arginine-L (D) -cysteine-L (D) -tetrapeptide (GCCR), glycine-L (D) -serine-L (D) -arginine-L (D) -L (SR), and other cysteine-containing peptides, including but not limited to Examples of the compound include one or more of 1- [ (2 s) -2-methyl-3-thiol-1-oxopropyl ] -L (D) -proline, thioglycolic acid, mercaptoethanol, thiophenol, D-3-troloxivirol and dodecylmercaptan.
2.5 identification of ligand-bound copper clusters
As an example, the following is characterization data for L-GSH-bound copper clusters (GSH-CuCs).
1) Morphological observation by Transmission Electron Microscopy (TEM)
The test powder (GSH-CuCs sample) was dissolved in ultrapure water to 2mg/L as a sample, and then the test sample was prepared by the pendant-drop method. The specific method comprises the following steps: 5 mul of the sample was dropped on a copper mesh, and the sample was naturally volatilized until the water drops disappeared, and then the morphology of the sample was observed by a JEM-2100F STEM/EDS field emission high resolution transmission electron microscope.
A and B panels of FIG. 1 show typical scanning electron microscope images of GSH-CuCs, and their size distribution was calculated from different transmission electron microscope images. The result shows that the GSH-CuCs have good dispersibility, and the size of the GSH-CuCs is between 0.5 and 5.0 nm.
2) X-ray photoelectron spectroscopy
X-ray photoelectron spectroscopy (XPS) was measured on ESCALAB 250xi X-ray photoelectron spectroscopy. Double-sided conductive adhesive (3 mm. Times.3 mm) was attached to an aluminum foil, and test powder was uniformly applied to the double-sided adhesive tape and covered with a layer of aluminum foil. The sample was held at 8 mpa pressure for 1 minute. The residual powder on the surface was removed and then a central sample (1 mm. Times.1 mm) was cut out for XPS testing.
The C-frame of FIG. 1 is XPS spectrum of copper element in GSH-CuCs. The two peaks appear at 931.98 and 951.88eV, respectively, which can be attributed to the 2p of copper 3/2 And 2p 1/2 Binding energy of electrons. No Cu 2p near 942.0eV 3/2 Satellite peaks confirmed the absence of Cu (II) electrons. Since the binding energy of Cu (0) is only 0.1eV away from that of Cu (I), it is impossible to exclude the formation of Cu (I), and the valence of copper in the resulting GSH-CuCs is likely to be between 0 and + 1.
3) Fourier transform infrared spectroscopy
The FT-IR spectra were tested on a PerkinElemer LS 55 fluorescence spectrometer. The test powder was dissolved in ultrapure water and measured at room temperature. The scanning range is 200-800nm, the sample cell is a standard quartz test tube, and the light path is 1cm.
Fig. 1, panel D, shows a comparison of FT-IR spectra of GSH-CuCs (top) and GSH (bottom). GSH exhibits a number of characteristic infrared bands, namely COOH (1390 and 1500 cm) -1 ) N-H stretching (3410 cm) -1 ) And NH2 group N-H bend (1610 cm) -1 ). At 2503cm -1 The peak observed at (a) can be attributed to the S-H tensile vibration mode. Stretching vibration band (2503 cm) except S-H -1 ) In addition, GSH-CuCs have corresponding infrared characteristics. The results indicate that the S-H bond is broken and the GSH molecule is bound to the surface of the copper core through the formation of copper-sulfur bonds.
4) Fluorescence spectroscopy
The test powder was dissolved in ultrapure water and measured by fluorescence spectrometry at room temperature.
As shown in panel E of fig. 1, at an excitation peak of 365nm, the copper cluster showed red emission with a peak at 595nm and a corresponding full width at half maximum (FWHM) of about 80nm. Notably, due to the aggregation-induced emission enhancement, the FL intensity of GSH-CuCs will increase significantly when ethanol is added to the solution. In addition, large stokes shifts (230 nm) show good promise for fluorescent probes and bioimaging.
EXAMPLE III animal experiments
3.1 Experimental materials and animals
3.1.1 test specimens
Cu-1: GSH modified copper cluster (GSH-CuCs) with size of 0.5-5nm.
Cu-2: cysteine (Cys) -modified copper clusters (Cys-CuCs) with a size of 0.5-5nm.
All test samples were synthesized as described above with minor modifications and their quality was assessed as described above.
3.1.2 Positive control samples
Sorafenib (Sorafenib).
3.1.3 test animals and groups
70 SPF male C57BL/6N mice (purchased from Beijing Huafukang laboratory animal technology Co., ltd. (production license number: SCXK (Jing) 2019-0008), 6-8 weeks old and 16-20g in weight are randomly divided into 7 groups (N = 10) according to the weight, namely a blank control group, a model group, a positive control group, a Cu-1 group low dose group, a Cu-1 high dose group, a Cu-2 low dose group and a Cu-2 high dose group.
3.2 modeling method
Except for the placebo group, the mice in each group were treated with carbon tetrachloride (CCl) 4 ) Preparing a liver cirrhosis model by an induction method. The molding method comprises (1) intraperitoneal injecting 10% CCl into each mouse according to 7 μ L/g body weight 4 (olive oil dilution) 2 times per week for 8 weeks; control mice were injected intraperitoneally with an equal amount of olive oil solvent. (2) Beginning at week 6 and 48h after the last weekly injectionSelecting 2 mice to be sacrificed, observing the appearance of the liver of the mice, and after the appearance accords with the characteristics of liver cirrhosis (8 th week), carrying out formalin fixation on liver tissues, and carrying out HE staining, massson staining and the like to evaluate the modeling condition of the liver cirrhosis model.
3.3 administration of drugs
After the model building is successful, feeding sorafenib 25mg/kg to the positive control group mice by intragastric administration; corresponding test articles are respectively given to the Cu-1, cu-2 low and high dose groups according to the intraperitoneal injection of 2.5 or 10mg/kg dose; the mice in the control group and the model group were administered with saline by intraperitoneal injection at a rate of 10 mL/kg. The administration is 1 time per day for 20 days.
3.4 Biochemical assays
After the administration, the mouse was subjected to orbital blood collection, serum collection, selection of a Zhongsheng north control kit, and 5 indexes of albumin (albumin, ALB), total bilirubin (TBil), alanine Aminotransferase (ALT), aspartate Aminotransferase (AST), and monoamine oxidase (MAO) by a biochemical analyzer (siemens). The detection method is carried out strictly according to the kit instructions.
Table 1 shows product information of the kit for biochemical detection
Figure BDA0003855305230000111
3.5 pathological examination
3.5.1 HE staining
After euthanasia of the mice, liver tissue samples of the mice were fixed by 4% paraformaldehyde fixing solution for more than 48 hours, and after fixation, gradient dehydration by alcohol and transparentization by xylene and ethanol were performed. The liver tissue was then waxed and embedded. And (3) after the embedded material is subjected to block trimming, block sticking and trimming, the liver tissue is sliced by a paraffin slicer, and the slice thickness is 4 mu m. HE staining main procedure was as follows: baking the slices in an oven at 65 ℃, and then carrying out xylene treatment and ethanol gradient dehydration on the slices. And dyeing with hematoxylin dyeing liquid, promoting blue turning of blue liquid and 0.5% eosin dyeing, treating the slices with gradient ethanol and xylene, and sealing the slices with neutral gum. Fibrotic changes in liver tissue were observed using a microscope.
3.5.2 Masson staining
The liver tissue slices of the mice are baked and dewaxed and dehydrated, and are nuclear-stained by Regaud hematoxylin staining solution after chromating treatment. After washing with water, the sections were stained with Masson ponceau red acid reddening solution, and the sections were rinsed with 2% glacial acetic acid aqueous solution and then differentiated with 1% phosphomolybdic acid aqueous solution. Directly dyeing with aniline blue or light green liquid, soaking and washing with 0.2% glacial acetic acid water solution for a moment, transparentizing with 95% alcohol, anhydrous alcohol and xylene, and sealing with neutral gum. The liver tissue was observed with a microscope.
3.6 results of the experiment
3.6.1 successful mold making
The liver of the mouse in the model group is divided into round or oval masses with different sizes by the hyperplastic septa, ALT, TBil and AST indexes in serum are obviously increased relative to a blank control group, ALB is obviously reduced relative to the blank control group, and MAO indexes have no obvious difference with the blank control group, but the numerical value is also increased. All the results prompt that the mold making of the experiment is successful.
3.6.2 Effect of test drugs on alanine Aminotransferase (ALT), total bilirubin (TBil), aspartate Aminotransferase (AST), monoamine oxidase (MAO) and Albumin (ALB)
As can be seen from FIG. 2A, the ALT activity in the model group showed a very significant increase (from an average of 43.5. + -. 8.1U/L to 188.5. + -. 4.9U/L, P < 0.01) relative to that in the blank control group, suggesting that liver function in the cirrhosis model mice was diseased. Compared with the model group, the low and high dose (the lowest 37.0 +/-5.7U/L and the highest 38.6 +/-5.6U/L) of Cu-1 and Cu-2 and the administration of a positive control (42.8 +/-5.4U/L) can significantly reduce ALT activity to the level of a blank control group (P < 0.01).
As can be seen from FIG. 2B, the AST activity of the model group showed a significant increase (from 141.8. + -. 13.5U/L to 192.0. + -. 11.3U/L, P < 0.05) relative to that of the blank control group. The AST activity can be obviously reduced to 146.3 +/-8.4U/L or 144.3 +/-8.1U/L by high-dose administration of Cu-1 and Cu-2 respectively, the AST activity is at the same level with a blank control group (141.8 +/-13.5U/L), and the AST activity is obviously different from that of a model control group (P is less than 0.01). Positive controls also reduced AST activity ((165.5 + -11.6U/L, P < 0.05), but at a lower magnitude than the high dose groups of Cu-1 and Cu-2.
As can be seen from FIG. 2C, the TBil concentration of the model group showed a significant increase (from 1.02. + -. 0.20. Mu. Mol/L to 2.91. + -. 0.39. Mu. Mol/L) relative to that of the blank control group, and was significantly different from that of the blank control group (P < 0.01). Compared with the model group, the serum TBil (the maximum is 1.16 +/-0.30 mu mol/L, the minimum is 1.08 +/-0.08 mu mol/L, and the P is less than 0.01) is remarkably reduced by the low dose and the high dose of Cu-1 and Cu-2, and the level is close to that of a blank control group (1.02 +/-0.20 mu mol/L).
As shown in FIG. 2D, the MAO activity in the model group was increased compared to that in the blank control group (18.8. + -. 2.9U/L for the blank control group and 21.5. + -. 0.7U/L for the model group), but there was no statistical difference, indicating that the carbon tetrachloride-induced change in the MAO activity index was not significant in the liver cirrhosis mice. However, compared with the model group, the high doses of Cu-1 and Cu-2 can respectively significantly reduce the serum MAO level to 17.3 +/-1.5U/L (P < 0.01) or 18.3 +/-2.1U/L (P < 0.05), and the effect is better than that of the positive control.
As can be seen from fig. 2E, the ALB level of the model group showed a significant decrease (from 24.2 ± 0.6g/L to 22.1 ± 1.3 g/L) relative to the ALB level of the blank group, which was significantly different from that of the blank group (P < 0.05), indicating that carbon tetrachloride treatment could significantly reduce serum ALB level. However, cu-1 and Cu-2 did not significantly affect serum ALB levels.
The results show that the ligand-bound copper cluster reduces ALT, AST, TBIL and MAO levels in a dose-dependent manner, and prompts the recovery of the liver function of a mouse, and the effect of the ligand-bound copper cluster is superior to that of a positive control medicament in at least some indexes
3.6.3 pathological assays
Cirrhosis is a pathological feature of diffuse fibrosis of liver tissue and the formation of pseudolobules. The results of HE staining pathological examination showed that, as shown in fig. 3A, the blank control group mice had clear normal liver tissue structure, intact liver lobules, aligned hepatocyte chords, radially arranged with central vein as the center, normal liver cell nuclei, and only a small amount of fibrous tissue in the region of the confluence; as shown in fig. 3B, in the liver of the model control group mouse, the arrangement of hepatocytes was disturbed, ballooning occurred, hepatic lobules were nearly disappeared, pseudolobules (right arrow in fig. 3B) appeared in large numbers, collagen fibers were proliferated in large numbers and formed round or oval fibrous spaces (left arrow in fig. 3B). Compared with the model control group, as shown in fig. 3C, the positive control group has obviously reduced liver damage degree, obviously tends to be neat in cell arrangement, collagen fibers are proliferated but obviously reduced, fibrous intervals are not formed, and false leaflets almost disappear; however, the gap between the liver cells of the positive control group was significantly increased compared to the normal liver tissue (downward arrow in fig. 3C). Compared with a model control group, the liver cells of the two copper cluster drug (Cu-1 and Cu-2) administration groups show extremely remarkable liver injury recovery phenomena, which are shown in that the fibroplasia and false lobule in the liver are obviously reduced, and certain dose dependence is shown.
Fig. 3D and 3E show HE staining patterns of the repair effect of low and high dose dosing on liver tissue damage, represented by Cu-1 copper clusters, respectively. As shown in fig. 3D, in the Cu-1 copper cluster low dose group, the hepatocytes were aligned, the pseudolobules were almost disappeared, the collagen fibril proliferation was also significantly decreased, and the hepatic cell gap was increased to some extent with respect to the normal liver tissue (indicated by the downward arrow in fig. 3D). As shown in fig. 3E, the Cu-1 copper cluster high dose administration group had a more significant improvement effect than the low dose administration group, the pseudolobules completely disappeared, the collagen fiber proliferation was not significant, the phenomenon of increase in the liver cell gap was hardly visible, and there was no significant difference from the normal liver cells. From this, it can be seen that the Cu-1 copper cluster drug exhibited better effect of repairing liver tissue damage than the positive control drug.
The Masson staining results were identical to the HE staining results.
The Cu-2 copper cluster drug also showed similar effects to the Cu-1 copper cluster drug.
In conclusion: the tested drugs of the Cu-1 and Cu-2 copper clusters obviously reduce liver fibroplasia and false lobule of liver. The liver function index test result also shows the phenomenon of liver function recovery. Of these, alanine Aminotransferase (ALT) and total bilirubin (TBil) are the most common. Aspartate Aminotransferase (AST) and monoamine oxidase (MAO) also recovered significantly, and the change in Albumin (ALB) was not significant. The two copper cluster test substances can obviously improve the liver function and the pathological structure of the liver cirrhosis mouse in the liver cirrhosis mouse, and the overall effect is superior to that of the positive control sorafenib, thereby providing experimental basis for further application in the future.
Different sizes of GSH-CuCs and Cys-CuCs and other ligand-bound CuCs were also tested using the same procedure, with similar results, and therefore will not be described in detail here.
While the invention has been described with reference to specific embodiments, it will be understood that the examples are illustrative and that the scope of the invention is not limited thereto. Alternative embodiments of the invention will become apparent to those of ordinary skill in the art to which the invention relates. Such alternative embodiments are to be considered as included within the spirit and scope of the present invention. The scope of the invention is, therefore, indicated by the appended claims, and is supported by the foregoing description.
Reference to the literature
Deng H.H.et al.An ammonia-based etchant for attaining copper nanoclusters with green fluorescence emission.Nanoscale,2018,10,6467.
Jia X.et al.Cu Nanoclusters with Aggregation Induced Emission Enhancement.Small,2013,DOI:10.1002/smll.201300896.
Wang C.and Huang Y.GREEN ROUTE TO PREPARE BIOCOMPATIBLE AND NEAR INFRARED THIOLATE-PROTECTED COPPER NANOCLUSTERS FOR CELLULAR IMAGING.NANO:Brief Reports and Reviews.2013,8(5):1350054(10 pages).

Claims (9)

1. Use of ligand-bound copper clusters for the manufacture of a medicament for the treatment of a patient with liver cirrhosis, wherein the ligand-bound copper clusters comprise:
a copper core; and
a ligand, wherein the ligand binds to the copper core forming a ligand-bound copper cluster;
the ligand is one selected from thymine, thymine Modified Hyaluronic Acid (TMHA), L-cysteine derivatives, D-cysteine derivatives, cysteine-containing oligopeptides and derivatives thereof, and other thiol-containing compounds.
2. Use according to claim 1, characterized in that the diameter of the copper core is between 0.5 and 5nm.
3. Use according to claim 1, characterized in that the diameter of the copper core is between 0.5 and 3nm.
4. Use according to claim 1, characterized in that said derivative of L-cysteine is selected from N-isobutyryl-L-cysteine (L-NIBC) and N-acetyl-L-cysteine (L-NAC) and said derivative of D-cysteine is selected from N-isobutyryl-D-cysteine (D-NIBC) and N-acetyl-D-cysteine (D-NAC).
5. Use according to claim 1, wherein the cysteine-containing oligopeptides and derivatives thereof are cysteine-containing dipeptides, cysteine-containing tripeptides or cysteine-containing tetrapeptides.
6. Use according to claim 5, characterized in that the cysteine-containing dipeptide is selected from the group consisting of L (D) -cysteine-L (D) -arginine dipeptide (CR), L (D) -arginine-L (D) -cysteine dipeptide (RC), L (D) -histidine-L (D) -cysteine dipeptide (HC) and L (D) -cysteine-L (D) -histidine dipeptide (CH).
7. Use according to claim 5, characterized in that the cysteine-containing tripeptide is selected from the group consisting of the glycine-L (D) -cysteine-L (D) -arginine tripeptide (GCR), the L (D) -proline-L (D) -cysteine-L (D) -arginine tripeptide (PCR), the L (D) -lysine-L (D) -cysteine-L (D) -proline tripeptide (KCP) and the L (D) -Glutathione (GSH).
8. Use according to claim 5, characterized in that the cysteine-containing tetrapeptide is selected from the group consisting of glycine-L (D) -serine-L (D) -cysteine-L-arginine tetrapeptide (GSCR) and glycine-L (D) -cysteine-L (D) -serine-L (D) -arginine tetrapeptide (GCSR).
9. Use according to claim 1, characterized in that the other thiol-containing compound is selected from 1- [ (2S) -2-methyl-3-thiol-1-oxopropyl ] -L (D) -proline, thioglycolic acid, mercaptoethanol, thiophenol, D-3-mercaptovaline, N- (2-mercaptopropionyl) -glycine and dodecyl mercaptan.
CN202211148299.6A 2020-03-16 2020-11-19 Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis Pending CN115444943A (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CN2020101819472 2020-03-16
CN202010181947 2020-03-16
CN202011306578.1A CN113398280B (en) 2020-03-16 2020-11-19 Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN202011306578.1A Division CN113398280B (en) 2020-03-16 2020-11-19 Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis

Publications (1)

Publication Number Publication Date
CN115444943A true CN115444943A (en) 2022-12-09

Family

ID=77677429

Family Applications (2)

Application Number Title Priority Date Filing Date
CN202211148299.6A Pending CN115444943A (en) 2020-03-16 2020-11-19 Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis
CN202011306578.1A Active CN113398280B (en) 2020-03-16 2020-11-19 Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis

Family Applications After (1)

Application Number Title Priority Date Filing Date
CN202011306578.1A Active CN113398280B (en) 2020-03-16 2020-11-19 Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis

Country Status (1)

Country Link
CN (2) CN115444943A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116328787A (en) * 2023-03-30 2023-06-27 武汉广行科学研究有限公司 Layered hydroxide supported gold nanoparticle and/or gold cluster composite material, preparation method and application thereof

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101074202B (en) * 2006-05-16 2012-08-08 重庆药友制药有限责任公司 Production of injecting reductive glutathione
JP2019533636A (en) * 2016-08-05 2019-11-21 深▲セン▼深見医薬科技有限公司Ｓｈｅｎｚｈｅｎ Ｐｒｏｆｏｕｎｄ−Ｖｉｅｗ Ｐｈａｒｍａ Ｔｅｃｈ Ｃｏ．， Ｌｔｄ． AuC-containing material, method for producing the same, and use thereof
CN114558029A (en) * 2019-01-16 2022-05-31 武汉广行科学研究有限公司 Copper cluster, thymine modified hyaluronic acid and poly-copper cluster, and preparation method and application thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116328787A (en) * 2023-03-30 2023-06-27 武汉广行科学研究有限公司 Layered hydroxide supported gold nanoparticle and/or gold cluster composite material, preparation method and application thereof

Also Published As

Publication number Publication date
CN113398280B (en) 2022-10-14
CN113398280A (en) 2021-09-17

Similar Documents

Publication Publication Date Title
Xiao et al. Dihydrolipoic acid–gold nanoclusters regulate microglial polarization and have the potential to alter neurogenesis
RU2304433C2 (en) Compositions inhibiting the development of blood vessels and method for their applications
CN113398280B (en) Ligand-bound copper clusters, compatibility of ligand-bound copper clusters and application of ligand-bound copper clusters in treatment of liver cirrhosis
Fan et al. Decoy Exosomes Offer Protection Against Chemotherapy‐Induced Toxicity
CN113398279B (en) Ligand-bound gold clusters, compositions and methods for treating liver cirrhosis
WO2021184762A1 (en) GOLD CLUSTERS (AuCs), COMPOSITION AND METHOD FOR TREATMENT OF LIVER CIRRHOSIS
Shan et al. RETRACTED ARTICLE: In vitro and in vivo protein release and anti-ischemia/reperfusion injury properties of bone morphogenetic protein-2-loaded glycyrrhetinic acid-poly (ethylene glycol)-b-poly (l-lysine) nanoparticles
AU2020436820B2 (en) Copper clusters, composition, and method for treatment of liver cirrhosis
RU2806633C1 (en) Copper clusters, composition and method of treatment of liver cirrhosis
RU2806634C1 (en) GOLD CLUSTERS (AuCs), COMPOSITION AND A METHOD OF TREATMENT OF LIVER CIRRHOSIS
JP7462744B2 (en) Compositions and methods for treating diabetes
CN107137419A (en) A kind of morindea officinalis polysaccharide extract is by stimulating GnRH releases effectively to recover the method for varicocele serum sex hormone level
Chiu et al. Development of a decellularized liver matrix-based nanocarrier for liver regeneration after partial hepatectomy
JP2023508454A (en) Compositions and methods for treatment of multiple sclerosis
WO2018033033A1 (en) Use of fullerene structure in preparation of medicament for treating anemia and complication thereof
CN112888700A (en) Modified nucleic acid having improved therapeutic efficacy and anticancer pharmaceutical composition comprising the same
RU2790828C1 (en) Hepatoprotective effect of low molecular weight mimetics ngf
TW201607955A (en) Polypeptide used for manufacturing multi-effect pharmaceutical composition of living body
CN109364093A (en) The inhibiting effect of the inhibitor in experimental Rat Hepatocellular Carcinoma of AKT
Hu et al. Curcumin-loaded graphene oxide quantum dots enhance otoprotective effects via blocking cuproptosis
CN112336874B (en) Nanometer medicine carrier and its prepn and application
CN114470158A (en) Application of bioactive peptide in medicine field
CN117752592A (en) Double-layer hydrogel microsphere and application thereof
Wang et al. Modulating macrophage phenotype for accelerated wound healing with chlorogenic acid-loaded nanocomposite hydrogel
US20070219126A1 (en) Class of bioactive glycoprotein

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 40087637

Country of ref document: HK