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Active targeting type ultrasonic/fluorescence bimodal contrast medium as well as preparation method and application thereof

A dual-modality contrast agent, contrast agent technology, applied in the fields of medicine and chemistry

Active Publication Date: 2017-10-27
NINGBO INST OF MATERIALS TECH & ENG CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few reports on the application of NPY receptor as a target in tumor imaging, especially in the field of ultrasound or fluorescence imaging.

Method used

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  • Active targeting type ultrasonic/fluorescence bimodal contrast medium as well as preparation method and application thereof
  • Active targeting type ultrasonic/fluorescence bimodal contrast medium as well as preparation method and application thereof
  • Active targeting type ultrasonic/fluorescence bimodal contrast medium as well as preparation method and application thereof

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preparation example Construction

[0100] The present invention also provides a method for preparing the contrast agent, the method comprising the following steps:

[0101] 1) providing a mixed solution comprising nano-microbubbles and target molecules;

[0102] 2) performing a coupling reaction between the target molecule and the nano-microbubble to obtain the contrast agent.

[0103] In another preferred example, the mass ratio of the target molecule to the nano-microbubble in step 1) is 1:3-30, preferably 1:5-20, more preferably 1:8-15.

[0104] In another preferred example, the nano-microbubbles in step 1) are pretreated by activation with an activator.

[0105] In another preferred example, the activator includes (but not limited to): N-hydroxysuccinimide, (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride salt), or a combination thereof.

[0106] In the present invention, the nano-microbubbles are prepared by the following method:

[0107] a-1) providing the first mixed solution and the seco...

Embodiment 1

[0149] Embodiment 1 Preparation of F-PLLA nano-microbubbles

[0150] (1) Take 0.1g of F-PLLA as a biodegradable polymer material, dissolve 60 μL of perfluorohexane in 4mL of dichloromethane as the inner aqueous phase, and ultrasonicate for 1 min in an ultrasonic cleaner to ensure that it is fully mixed in the solvent uniform.

[0151] (2) In the centrifuge tube of 50mL, add the ultrapure water of 20mL and 0.3g emulsifying agent sodium cholate successively, in ultrasonic cleaning machine, ultrasonic 5min guarantees that it fully dissolves, obtains the sodium cholate solution of about 1.5wt% (per 100ml The solution contained 1.5 g sodium cholate).

[0152] (3) Add the organic solution prepared in step (1) and the sodium cholate solution prepared in step (2) into a beaker, and disperse it evenly under the condition of a high-speed homogenizer at 10,000 rpm. Under the ultrasonic cell pulverizer, the power of 643W was sonicated for 2min.

[0153] (4) Stir the solution obtained i...

Embodiment 2

[0160] Example 2 F-PLLA nanobubble surface coupling [Pro30, Nle31, Bpa32, Leu34] NPY (28-36) (PNBL-NPY)

[0161] (1) 10 mg of the sample obtained in Example 1 was made into 10 mg mL -1 PBS solution, respectively add 1ml 3mg mL -1 N-hydroxysuccinimide solution with 3mg mL -1 (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) solution activates the amino groups on the surface of the microbubbles.

[0162] (2) The concentration of 1ml PNBL-NPY is 0.8mg mL -1 Add the PBS solution obtained in step (1) into the activated solution, stir evenly at room temperature for 20 hours, and use the carboxyl group of PNBL-NPY to condense with the amino group on the surface of the microbubble to obtain the required active targeting nano-microbubble (PNBL-NPY-F-PLLA nanobubbles).

[0163] (3) Centrifuge the sample obtained in step (2) for half an hour at 12000 g at 4 degrees Celsius, remove the supernatant, and store it for future use.

[0164] figure 2 It is the TEM test result...

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Abstract

The invention relates to an active targeting type ultrasonic / fluorescence bimodal contrast medium as well as a preparation method and application thereof. Specifically, the contrast medium comprises nano microbubbles and targeting molecules, wherein the nano microbubbles are filled with an inner water phase; and the targeting molecules are coupled with the surfaces of the nano microbubbles. The invention further discloses a preparation method and application of the contrast medium. The contrast medium is excellent in biocompatibility and relatively small in particle size, and can not only exist and be used stably, but also effectively permeate into tumor cells, and thus targeting radiography upon tumor cells is achieved. The preparation method is simple in process and economic and safe.

Description

technical field [0001] The invention relates to the fields of medicine and chemistry, in particular to an active-targeting ultrasound / fluorescence dual-mode contrast agent and its preparation method and application. Background technique [0002] Imaging diagnosis has become a common means of tumor detection due to its intuitive and efficient characteristics. Commonly used medical imaging diagnostic techniques include: ultrasound imaging (US), magnetic resonance imaging (MRI), positron emission tomography (PET), fluorescence imaging, and X-ray computed tomography (CT). Compared with other medical imaging technologies, ultrasound is a non-invasive, low-cost, real-time imaging technology, and is currently the most widely used imaging method in clinical medicine. However, the image resolution and sensitivity of traditional ultrasound diagnostic techniques are low, and the use of ultrasound contrast agents can greatly improve the contrast between detected tissues and normal tiss...

Claims

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Application Information

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IPC IPC(8): A61K49/22A61K49/00
CPCA61K49/0019A61K49/0093A61K49/221A61K49/223
Inventor 李娟田宇辰吴爱国
Owner NINGBO INST OF MATERIALS TECH & ENG CHINESE ACADEMY OF SCI
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