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Nano-gold CT contrast agent for early diagnosis of hepatocellular carcinoma and preparation method of contrast agent

An early diagnosis and nano-gold technology, applied in the field of biomedicine, can solve the problems of unsuitable animal CT experiments and poor stability, and achieve the effects of improving enrichment and tissue penetration, improving stability, and good biocompatibility

Inactive Publication Date: 2016-07-27
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the system has poor stability at the concentration (0.1M) required for CT contrast, and is not suitable for animal CT experiments.

Method used

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  • Nano-gold CT contrast agent for early diagnosis of hepatocellular carcinoma and preparation method of contrast agent
  • Nano-gold CT contrast agent for early diagnosis of hepatocellular carcinoma and preparation method of contrast agent
  • Nano-gold CT contrast agent for early diagnosis of hepatocellular carcinoma and preparation method of contrast agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Preparation of nano-gold CT contrast agent for early diagnosis of liver cancer

[0046] 1) LA-PEG 2k Synthesis: Weigh 2 g of single-ended methoxy mPEG 2k -OH and 0.206g lipoic acid (LA) (the molar ratio is LA:mPEG 2k -OH=5:1) was added to a 50mL round bottom flask, and 960mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC.HCL) and 610mg of 4-bis Dimethylaminopyridine (DMAP), dissolved in 30 mL of dichloromethane (DCM). Stir in an ice-water bath for 30 minutes, then react in a water bath at 35°C for 2 days. After the reaction, dichloromethane was removed by rotary evaporation. The remaining solid was dissolved in distilled water, transferred to a dialysis bag (MWCO 2k) and dialyzed for two days. lyophilized to obtain LA-PEG 2k White powdery solid. Characterized by NMR, see attached image 3 .

[0047] 2) LA-PEG 4k and LA-PEG 10k The synthesis of: method is the same as step 1.

[0048] 3) Preparation of AuNPs: Weigh 59 mg tetrachlo...

Embodiment 2

[0058] Example 2: Nano-gold CT contrast agent ligand shielding-deshielding investigation:

[0059] 1) Assembly-disassembly investigation of nano-gold CT contrast agent

[0060] For nano gold CT contrast agent, its assembly-disassembly properties under normal tissue (pH7.4) and tumor conditions (pH6.8) were investigated. In the experimental group 1:4 (see attached figure 2 ) and control group 1 as an example, the details are as follows: Add NaOH solution (pH13) dropwise to the nano-gold solution to pH8-9. Solutions HEPESpH7.4 and HEPESpH6.8, use dynamic light scattering (DLS) to investigate the particle size at different pHs, the particle size of the assembly is 100-200nm, and the particle size of the disassembly is 30-40nm, which is used to illustrate the assembly-disassembly of the system nature. Then the pH of the solution was adjusted repeatedly between pH 7.4 and pH 6.8, and the particle size change of the test system was used to investigate the pH-sensitive reversible...

Embodiment 3

[0066] Example 3: Investigation on Concentrated Stability of Nano-gold CT Contrast Agent:

[0067] Batch centrifugation is used to concentrate the modified nano-gold solution (ie nano-gold CT contrast agent). For the first centrifugation (25,000 rpm, 45 minutes), the batch of nano-gold liquid is concentrated about 20 times; for the second centrifugation (15,000 rpm, 60 minutes), the concentrated solution obtained by the first centrifugation is further concentrated about 8 times. Finally, the nano-gold concentrated solution (0.15M) with a concentration (0.1M) suitable for CT imaging of tumor-bearing mice was obtained. Precipitation occurred in the experimental group 1:0 and the experimental group 1:2, which indicated that the contrast agent had poor stability at the contrast concentration and was not suitable for in vivo CT imaging. The test results are attached Figure 10 .

[0068] Combining Example 2 and Example 3, it is confirmed that the gold nano CT contrast agent suit...

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Abstract

The invention discloses a nanogold CT contrast agent for early diagnosis of hepatocellular carcinoma and a preparation method of the contrast agent. PH-sensitive lipoic acid modified N-diisopropyl ethanediamine (LA-N(iPr)2), lipoic acid polyethylene glycol modified glycyrrhetinic acid (LA-PEG3.5k-GA) and lipoic acid linked polyethylene glycol (LA-PEG2k) are modified on the surface of nano-gold (AuNPs), and the mass ratio of the n(LA-N(iPr)2) to the n(LA-PEG3.5k-GA) to the n(LA-PEG2k) is at 1 to 1 to (3-5). The contrast agent has a pH-sensitive ligand reversible shielding-deshielding function, and the contrast agent, which combines active targeting and passive targeting, can achieve an imaging effect which is more excellent than that of the active targeting. Based upon mouse CT imaging experiments, the contrast agent is applicable to CT imaging of subcutaneously hetetotopic tumor and hepatocellular carcinoma in situ in tumor-bearing mice (mouse H22 hepatocellular carcinoma).

Description

technical field [0001] The invention provides a nano-gold CT contrast agent and a preparation method thereof. The contrast agent prepared by the method is suitable for early CT detection of liver cancer and belongs to the technical field of biomedicine. Background technique [0002] At present, the mortality rate of primary liver cancer in the world ranks third among malignant tumors, and the incidence and mortality rate of liver cancer in my country account for more than half of the world (L.A.Torre, F.Bray, R.L.Siegel, J.Ferlay, J.Lortet - Tieulent, A. Jemal, Global Cancer Statistics, 2012, CA. Cancer J. Clin. 65 (2015) 87–108. doi:10.3322 / caac.21262.). There are two main reasons for the high mortality rate of liver cancer: first, it is difficult to diagnose early, and liver cancer is a deep tumor (2.5-10 cm in depth), which often exists in the deep part of the liver. Once discovered, it is often in the middle and late stage; The effect of chemotherapy is poor, and the dos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/04
CPCA61K49/04
Inventor 袁直王蔚呼振鹏田志清马金龙
Owner NANKAI UNIV
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