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Compound for preventing osteonecrosis and/or promoting angiogenesis and pharmaceutical application thereof

A compound and drug technology, applied in the field of biomedicine, can solve problems such as limited effects and failure to meet clinical needs, and achieve the effects of promoting proliferation and migration, good application prospects, and promoting angiogenesis

Pending Publication Date: 2022-07-22
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effect of DNA tetrahedral framework nucleic acid in preventing bisphosphonate-related osteonecrosis of the jaw is limited and cannot meet the clinical needs

Method used

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  • Compound for preventing osteonecrosis and/or promoting angiogenesis and pharmaceutical application thereof
  • Compound for preventing osteonecrosis and/or promoting angiogenesis and pharmaceutical application thereof
  • Compound for preventing osteonecrosis and/or promoting angiogenesis and pharmaceutical application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1. Synthesis of the complex of tFNA and KLT polypeptide (tFNA-KLT)

[0036] refer to figure 1 As shown in the schematic diagram, the complex tFNA-KLT of tFNA and KLT is synthesized, and the specific steps are as follows:

[0037] (1) Synthesis of tFNA

[0038] The four DNA single strands (S1, S2, S3, S4) were dissolved in TM buffer (10 mM Tris-HCl, 50 mM MgCl 2 , pH=8.0), control the final concentration of the four DNA single strands to 1000nM, mix well, rapidly heat to 95°C for 10 minutes, and then rapidly cool down to 4°C for 20 minutes to obtain DNA tetrahedral framework nucleic acid: tFNA.

[0039] (2) Synthesis of complex tFNA-KLT

[0040] Then, according to the molar ratio of tFNA:KLT=1:100, tFN and KLT were co-incubated at room temperature for 30 min to obtain a complex of tFNA and KLT polypeptide: tFNA-KLT.

[0041] The sequence of the four single-stranded DNA (5'→3') is as follows:

[0042] S1:

[0043] ATTTATCACCCGCCATAGTAGACGTATCACCAGGCAGTTGAGACGA...

experiment example 1

[0053] Experimental example 1. Screening experiment of the ratio of tFNA and KLT

[0054] 1. Experimental method

[0055] The complex of tFNA and KLT was prepared by referring to the method of Example 1, the only difference being that the molar ratio of tFNA:KLT in the control step (2) was 1:1, 1:10, 1:50, 1:100, 1:150, respectively , 1:200, 1:300. The complexes of tFNA:KLT co-incubated with different molar ratios were obtained.

[0056] Identification: The complexes obtained after co-incubating tFNA and KLT with different molar ratios were detected by PAGE electrophoresis; the shape of tFNA-KLT was detected by atomic force microscopy; the zeta potential and particle size of tFNA-KLT were detected by dynamic light scattering.

[0057] 2. Experimental results

[0058] like figure 2 As shown, the electrophoresis results showed that when the molar ratio of tFNA:KLT was 1:100, the maximum ratio of tFNA loaded with KLT was reached, so the optimal molar ratio of tFNA and KLT co...

experiment example 2

[0060] Experimental example 2. Uptake of tFNA-KLT by human umbilical vein endothelial cells (HUVEC)

[0061] 1. Experimental method

[0062] FAM (6-carboxyfluorescein)-modified KLT and tFNA-KLT were mixed in the cell culture medium at a concentration of 250 nm, and then cultured for 6 h. To take images, HUVECs were washed with phosphate buffered saline (PBS) and fixed with 4% paraformaldehyde. Cytoskeleton was stained with phalloidin and nuclei were stained with DAPI.

[0063] The tFNA-KLT used in this experimental example was prepared in Example 1.

[0064] 2. Experimental results

[0065] like Figure 4 As shown, the results of immunofluorescence staining showed that both tFNA-KLT and KLT were taken up by HUVECs at 6 h; compared with KLT, the uptake effect of tFNA-KLT was better.

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Abstract

The invention provides a compound for preventing osteonecrosis and / or promoting angiogenesis and pharmaceutical application thereof, and belongs to the field of biological medicine. The compound is prepared by taking DNA tetrahedral framework nucleic acid and KLT polypeptide as raw materials, and the molar ratio of the DNA tetrahedral framework nucleic acid to the KLT polypeptide is 1: (1-300). Experimental results show that the compound can effectively promote proliferation and migration of endothelial cells, promote vascularization and play a good role in preventing bisphosphonate-related jaw necrosis (BRONJ). It is worthy of noting that compared with DNA tetrahedral framework nucleic acid or KLT polypeptide, the compound has the advantages that the effects of promoting proliferation and migration of endothelial cells and promoting angiogenesis are remarkably improved, and the synergistic interaction effect is achieved. The compound provided by the invention can be used for preparing a medicine for promoting angiogenesis and a medicine for preventing osteonecrosis, and has a very good application prospect.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a compound for preventing osteonecrosis and / or promoting angiogenesis and its pharmaceutical use. Background technique [0002] Osteonecrosis refers to a series of complex pathological processes in which the bone circulation is interrupted, the active components of the bone die, and subsequently repaired due to various reasons (mechanical, biological, etc.). Osteonecrosis can affect many joints throughout the body. Common osteonecrosis includes necrosis of the femoral head, osteonecrosis of the jaw, femoral condyle necrosis, and humeral head necrosis. [0003] Bisphosphonates (BPs), as a kind of anti-bone resorption drugs, are mainly used in the prevention of osteoporosis, Paget disease (Paget's disease, also known as eczema-like carcinoma), and multiple myeloma. Treatment and prevention of tumor bone metastases. However, long-term use of BPs will cause a serious adverse reaction - B...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/10A61K31/713A61P19/08A61P43/00
CPCA61K38/10A61K31/713A61P19/08A61P43/00A61K2300/00
Inventor 林云锋赵丹蔡潇潇肖德昡
Owner SICHUAN UNIV
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