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Application of Purmorphamine in pharmacy

A drug and a technology for inhibiting osteoclasts, which can be used in the pharmaceutical field to solve problems such as the influence of osteoclasts that has not yet been studied.

Inactive Publication Date: 2019-11-05
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In previous literature reports, Purmorphamine can significantly promote osteoblast differentiation, but its effect on osteoclasts, another important component of bone metabolism, has not yet been studied.

Method used

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  • Application of Purmorphamine in pharmacy
  • Application of Purmorphamine in pharmacy
  • Application of Purmorphamine in pharmacy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Purmorphamine inhibits osteoclast differentiation without significant cytotoxicity

[0028] (1) Isolation and culture of primary bone marrow-derived monocyte-macrophage BMMs

[0029] Take a C57BL / 6 mouse aged 5 to 8 weeks, put it to death by pulling its neck, soak it in 75% medical alcohol for 3 minutes, dissect the lower limbs of the mouse in a clean workbench, separate the skin, fascia, and knee and hip joint capsules, The mouse femur and tibia were obtained and placed in a bacterial culture dish previously added with sterilized phosphate-buffered saline. Use sterile scissors to cut the upper end of the femur and the lower end of the tibia, separate the bone marrow by centrifugation at 10,000 rpm, add 2ml of red blood cell lysate to resuspend the bone marrow, and let stand for 5 minutes to lyse the red blood cells. Then centrifuge horizontally at room temperature (1000rpm, 5 minutes), discard the liquid, add 10ml of complete Alpha MEM culture solution cont...

Embodiment 2

[0040] Example 2: Purmorphamine activates Gli1, an important transcription factor in the Hedgehog signaling pathway, and inhibits the expression of related characteristic genes in the process of osteoclast differentiation

[0041] (1) Cell treatment and RNA collection and separation

[0042] A control group and 3 drug-treated groups were set up, with 3 replicate wells in each group, and mononuclear macrophages were planted in a 6-well plate (40×10 4 Cells / well), the next day the culture medium was replaced with complete Alpha MEM culture medium containing 50ng / ml RANKL and 30ng / ml M-CSF, and 0.5, 1, 2μM Purmorphamine were added to the drug treatment group, and the control group was added with In the 2 μM Purmorphamine group, the same amount of dimethyl sulfoxide solvent was changed every other day, and the culture was continued for 6 days. When the control group had obvious mature and hypertrophic osteoclasts, it was terminated. Use the RNA extraction kit to extract the total...

Embodiment 3

[0045] Example 3: Purmorphamine inhibits the activation of the JNK signaling pathway and the expression of important transcription factors NFATc1 and c-fos during osteoclast differentiation

[0046] (1)Protein sample processing and extraction

[0047] Short-term protein treatment: seed monocyte-macrophages in 6-well plates (40×10 4 Cells / well, 30ng / ml M-CSF complete Alpha MEM culture medium), divided into control group drug treatment group, add 2μM Purmorphamine to the drug treatment group respectively 4 hours before protein extraction on the next day, and add the same amount of dimethylformamide to the control group Pretreatment with sulfoxide. Both groups were added with 50ng / ml RANKL to activate osteoclast differentiation-related signaling pathways, and protein samples were collected at 0, 5, 15, 30, and 60 minutes, respectively.

[0048] Long-term protein treatment: seed monocyte-macrophages in 6-well plates (40×10 4 cells / well, 30ng / ml M-CSF complete Alpha MEM culture ...

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Abstract

The invention discloses an application of Purmorphamine in pharmacy, which includes an application in preparation of a medicine for inhibiting differentiation and maturation of osteoclasts. The application found that Purmorphamine can inhibit the differentiation of osteoclasts, can be applied to the prevention and treatment of osteolysis mediated by titanium metal particles, and can play a more significant role.

Description

technical field [0001] The present invention relates to the use of Purmorphamine, in particular to the use in the field of pharmacy. Background technique [0002] my country's aging population is becoming more and more serious. At present, my country's population over 60 years old has exceeded 210 million, and it is expected to increase to 355 million by 2030 and 410 million by 2040. The health problems related to the aging population are very serious. Among them, bone and joint diseases seriously affect the survival time and quality of patients. Studies have shown that within 1 year after hip fracture, about 50% of patients will become disabled, and 20% of patients will die of various complications. The quality of life of patients is significantly improved. decline. Total joint arthroplasty (TJA) uses surgical techniques to implant artificial joint prostheses made of metal, high molecular polyethylene or ceramics into the body to replace the support and movement functions ...

Claims

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Application Information

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IPC IPC(8): A61K31/5377A61P19/08A61P19/02A61P29/00A61P19/10A61P35/00A61P35/04
CPCA61K31/5377A61P19/08A61P19/02A61P29/00A61P19/10A61P35/00A61P35/04
Inventor 陈建权章礼炜杨彦军
Owner SUZHOU UNIV
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