Drugs contg. as active ingredient midkine or inhibitors thereof

A technology of active ingredients and inhibitors, which is applied in the field of pharmaceuticals that control the function of neutrophils, and can solve problems such as toxicity

Inactive Publication Date: 2001-08-01
MEDICAL THERAPIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the activated inflammatory system is toxic to normal tissues and cells, and the strongly expressed inflammatory response should be suppressed

Method used

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  • Drugs contg. as active ingredient midkine or inhibitors thereof
  • Drugs contg. as active ingredient midkine or inhibitors thereof
  • Drugs contg. as active ingredient midkine or inhibitors thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Detection of midkine(MK) by ELISA

[0043] Synovial fluid samples were collected by aspiration from patients with inflammatory synovitis in OA or RA (age range 26 to 72; mean age 52 years). MK was detected in synovial fluid samples by ELISA (Muramatsu, H et al., J. Biol. Chem., 119:1171-1175, 1996). MK was undetectable in any synovial fluid samples from 3 healthy subjects, but was detected in all samples from 6 RA patients ( figure 1 , MK concentration is 62 to 10,000 pg / ml). Significant amounts of MK were detectable in 4 of 6 samples each from a different OA patient ( figure 1 , below the detection limit, to 1225pg / ml). The results indicated that MK levels in synovial fluid were significantly correlated with the inflammatory state of synovitis.

Embodiment 2

[0045] Immunohistochemical detection of MK

[0046]Synovial tissues were obtained from whole knee sections of 3 RA patients and 2 OA patients. All biopsy samples contained proliferative inflammatory periosteal tissue, histologically characterized by proliferation of cells lining the synovium, extensive infiltration of lymphocytes and macrophages, and profuse vascularization. Immunohistochemical detection was performed using the method of Muramatsu et al. (Muramatsu, H et al., Developmental Biology, 159:392-402, 1993). Biopsy samples were fixed in neutral-buffered formalin, embedded in paraffin, and cut into 5 μm thick sections. Sections were incubated overnight at 4°C with anti-human MK antibody (15 mg / ml) in PBS containing 0.2% bovine serum albumin and 2% normal goat serum. Using rabbits immunized with chemically synthesized human MK purchased from Peptide Institute, anti-human MK antibody was prepared by the method of Muramatsu et al. Control samples were incubated with P...

Embodiment 3

[0049] Detection of MK by Western blot analysis

[0050] Synovial tissue extracts were subjected to Western blot analysis, samples were electrophoresed on SDS-polyacrylamide gels by the method of Laemmli (Laemmli, U.K., Nature 227:680-685, 1970), and samples were electrophoresed on SDS-polyacrylamide gels by Towin et al. Proc.Natl.Acad.Sci.USA, 76:4350-4354,1979) method to transfer the protein in the gel to the nitrocellulose membrane. Nitrocellulose membranes were incubated overnight at 4°C with Dulbecco's phosphate-buffered saline (PBS) containing 5% skim milk. Then it was incubated with diluted anti-human MK antibody (diluted to 20 mg / ml with 5% skim milk solution) for 2 hours at room temperature. The nitrocellulose membrane was washed with PBS containing 0.1% Tween 20, incubated with an affinity-purified anti-rabbit IgG-horseradish peroxidase conjugate (Jackson Immunoresearch Laboratories, Baltimore, USA), and incubated with 4-chloro - 1-naphthol staining.

[0051] High...

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Abstract

Drugs for treating neutrophilic functional disorders or inflammatory diseases, containing as the active ingredient midkine (MK), which is a retinoic acid-inducible heparin-binding growth factor found to enhance the neutrophile migration and to exist at a high concentration in an inflammatory state, or inhibitors thereof.

Description

technical field [0001] The present invention relates to an agent for controlling neutrophil function, which contains midkine or an inhibitor thereof as an active ingredient. The agent can be used to treat diseases associated with neutrophils, such as neutrophil dysfunction or inflammatory diseases. Background technique [0002] Neutrophils are a type of granulocytes that have migratory, phagocytic and microbicidal activities and play an important role in biological protection against infectious agents such as bacteria or fungi. There are some diseases that are associated with neutrophils. [0003] Neutrophil dysfunction (in which one or more of the above biological activities of neutrophils is impaired) includes, for example: Lazy-leukocyte (chemotactic-defective leukocyte) syndrome (a neurotropic-deficiency disorder ). In patients with this syndrome, the number of neutrophils in the peripheral blood is significantly reduced, their migratory activity is impaired, but the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/18A61K38/19
CPCA61K38/18A61P19/00A61P19/02A61P29/00A61P43/00A61K38/17
Inventor 高田彻鸟山和宏村松寿子村松乔
Owner MEDICAL THERAPIES
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