A kind of dna nano-medicine targeting fat cells and its preparation method and application
A technology of adipocytes and nano-drugs, which is applied in the direction of drug combinations, pharmaceutical formulas, and medical preparations with non-active ingredients, etc., can solve the problems of threatening the life safety of patients, poor treatment effect, and treatment failure, and achieve good treatment effect, High load capacity, less side effects
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0036] The preparation process diagram of the DNA nanomedicine of the present invention, such as figure 1 shown, as follows:
[0037] 1. Ligation: Phosphorylated DNA template (10 μM, 0.6 μL) (shown in SEQ ID NO: 2) and primers (10 μM, 1.2 μL) (shown in SEQ ID NO: 3) containing the targeting sequence of adipocyte nucleic acid aptamer (MA-91) ) in 10 μL of DNA ligase buffer (5 mM Tris-HCl, 1 mM MgCl 2 , 0.1 mM ATP and 1 mM dithiothreitol) for annealing (keep at 95 °C for 5 min, then slowly cool down to room temperature within 2 h). The annealed product was ligated with Ecoli DNA ligase (60 U / μL, 1.6 μL) at 16° C. for 3 h to form a circularized DNA template.
[0038] 2. Polymerization: Circularized DNA template (0.6 μM, 5 μL) with phi29 DNA polymerase (10 U / μL, 2 μL), dNTPs (10 mM / μL, 2 μL) and BSA (0.05%, 0.1 μL) in 10 μL polymerase buffer solution (50 mM Tris-HCl, 10 mM (NH 4 ) 2 SO 4 , 10mM MgCl 2 and 4 mM dithiothreitol), after polymerization at 30 °C for 24 h, the rea...
Embodiment 2
[0049] Example 2 Study on the biological properties of TADN
[0050] 1. The scavenging of reactive oxygen species and reactive nitrogen species (RONS) by TADN
[0051] With the excessive absorption of nutrients by the body, excessive RONS is continuously produced and accumulated in the body, which will lead to oxidative stress in the body, and then lead to related metabolic inflammation, insulin resistance and other complications. Therefore, the clearance of RONS may be an innovative, versatile, safe and effective therapeutic strategy in obesity intervention, which requires various RONS-eliminating antioxidant drugs with broad antioxidant activities. However, the shortcoming of the narrow antioxidant activity range of traditional antioxidants greatly limits their biological applications and seriously affects their therapeutic effects on diseases. Therefore, an ideal antioxidant needs to have the characteristics of safety, targeting, strong antioxidant capacity, good biocompat...
Embodiment 3
[0079] Example 3 Animal experimental research
[0080] In vivo experiments
[0081] ①In vivo experiment and establishment of obesity model
[0082] C57BL / 6J male mice were purchased from Hunan Slike Jingda Laboratory Animal Co., Ltd. Animal experiments were approved by the Laboratory Animal Management Center of Hunan University. To establish an obesity model, 6-week-old mice were fed a high-fat diet (HFD, 60% calories, penSourceDiet D12492, Research Diet). Mice were weighed weekly until fed to about 30 g.
[0083] ②Biodistribution of TADN
[0084] After the mice were fed a high-fat diet to ~30 g, normal C57BL / 6 mice or obese mice were injected with Cy7-TADN or Cy7-NTADN through the tail vein. Then, the mice at different time points were imaged with the IVIS Lumina II Intravital Imaging System (Caliper LifeScience, USA). After 24 h, mice were sacrificed and major organs (heart, liver, spleen, lung and kidney) were collected and imaged.
[0085] ③The in vivo anti-obesity ...
PUM
| Property | Measurement | Unit |
|---|---|---|
| size | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More