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An ultrasonic prostate cancer diagnosis targeting reagent and a preparing method thereof

A targeting reagent and technology for prostate cancer, applied in echo/ultrasonic imaging agents, etc., to achieve improved imaging effects, enhanced imaging effects, and excellent targeting effects

Active Publication Date: 2014-08-13
YUEYANG INTEGRATED TRADITIONAL CHINESE & WESTERN MEDICINE HOSPITAL SHANGHAI UNIV OF CHINESE TRADITIONAL MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on such targeted ultrasound contrast agents

Method used

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  • An ultrasonic prostate cancer diagnosis targeting reagent and a preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preparation of EpDT3-SH Modified Targeting Reagent for Ultrasound Diagnosis of Prostate Cancer (1)

[0036] 1. Preparation of thiol-modified EpDT3 aptamer (EpDT3-SH)

[0037] Take 0.1 μmol sulfhydryl-protected EpDT3 aptamer (the specific sequence composition of sulfhydryl-protected EpDT3 aptamer is 5'-GCGACUGGUUACCCGGUCG-(CH 2 )6- S- S- (CH 2 )6-OH- 3'), dissolved in 2.5ml 0.1mol / l dithiothreitol (DTT) solution (pH 8.0), stirred at room temperature for 30 minutes, passed through Glen Gel-Pak? 2.5 desalting column (Glen Research, Sterling, VA) to remove unreacted DTT to obtain pure thiol-modified EpDT3 aptamer. The specific sequence composition of the sulfhydryl-modified EpDT3 aptamer is 5'-GCGACUGGUUACCCGGUCG- (CH 2 )6-SH- 3', and has 2'-fluoropyrimidine modification. The nucleotide sequence number of the EpDT3 aptamer is shown in SEQ ID NO.1.

[0038] 2. Synthesis of nanocarrier material PLGA-PEG-MAL

[0039] Take 5g of PLGA-COOH (glycolide ratio: 75 / 25...

Embodiment 2

[0044] Example 2 Preparation of EpDT3-SH Modified Prostate Cancer Targeting Reagent for Ultrasound Diagnosis (2)

[0045] 1. Preparation of thiol-modified EpDT3 aptamer

[0046] Concrete method is with embodiment 1.

[0047] 2. Synthesis of nanocarrier material PLGA-PEG-MAL

[0048] Dissolve 5g of PLGA-COOH (glycolide ratio: 75 / 25) in 20ml of dichloromethane, add 50mg of condensing agent DCC (dicyclohexylcarbodiimide), 20mg of TEA (triethylamine) and 30mg of NHS, mix Uniform, react at room temperature for 24h; then add 560mg NH 2 -PEG-MAL and 150 μL acid-binding agent DIEA (N,N-diisopropylethylamine) continued to react for 12 hours, and after the reaction was completed, use a rotary evaporator to remove dichloromethane under reduced pressure; add DMF ( N,N -dimethylformamide) was dissolved, and the solution was dialyzed with a dialysis bag to remove small molecular impurities and unreacted PEG, and freeze-dried to obtain the nanocarrier material PLGA-PEG-MAL. The results o...

Embodiment 3

[0053] Example 3 Preparation of EpDT3-SH Modified Targeting Reagent for Ultrasonic Diagnosis of Prostate Cancer (3)

[0054] 1. Preparation of thiol-modified EpDT3 aptamer

[0055] Concrete method is with embodiment 1.

[0056] 2. Synthesis of nanocarrier material PLGA-PEG-MAL

[0057] Concrete method is with embodiment 1.

[0058] 3. Preparation of perfluorooctyl bromide nanoparticles

[0059] Add 20mg PLGA-PEG-MAL and 12μL PFOB into 0.4ml chloroform solution, then add 2 mL of 1% polyvinyl alcohol (PVA) aqueous solution, mix in an ice-water bath with a homogenizer for about 60 s, and place the dispersion in an ice-water bath Use 200W power for intermittent ultrasonication for 180 s, and then use a homogenizer for 300 s. The obtained nanoemulsion was stirred at 20° C. for about 3 hours to volatilize the chloroform. The particle size of perfluorooctyl bromide nanoparticles in the obtained nanoemulsion was determined by laser light scattering to be 110 nm, and the encapsu...

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Abstract

The invention relates to an ultrasonic prostate cancer diagnosis targeting reagent and a preparing method thereof. The targeting reagent is perfluoro bromooctane nanoparticles modified by an EpDT3 aptamer. The EpDT3 aptamer is selected from: a) EpDT3-SH that is 5'-GCGACUGGUUACCCGGUCG-(CH2)6-SH-3' and is modified by 2'-fluoropyrimidine; or b) EpDT3-NH2 that is 5'-NH2-spacer-GCGACUGGUUACCCGGUCGinvertedT-3' and is provided with 2'-fluoropyrimidine and 3'-inverted T, wherein the 5'-amino is connected through hexaethylene glycol. The targeting reagent modified by the EpDT3 aptamer can be retained on tumor vascular beds and has excellent targeting property and significantly enhanced developing effects.

Description

Technical field [0001] The invention involves the targeted reagent used in ultrasound diagnosis. Specifically, it involves a prostate cancer ultrasound diagnostic targeting reagent and preparation method. Background technique [0002] Ultrasonic examination is the first choice and important imaging examination of prostate diseases, but currently all imaging examinations cannot be sensitive to chronic prostatitis, prostate hypertrophy, and early prostate cancer.Guide the prostate more puncture biopsy.In recent years, the performance of ultrasonic instruments has improved, and the emergence of new generation of acoustic film creation has created a new application area for ultrasound diagnosis.At the same time, the CEUS (CEUS) has been further developed in the field of diagnosis, and has become one of the non -invasive imaging methods for diagnosis of prostate cancer in the field of diagnosis.Studies have proved that the two can effectively enhance the two -dimensional ultrasound im...

Claims

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Application Information

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IPC IPC(8): A61K49/22
Inventor 朱全刚陈中建童仙君徐蓓蕾张若曦
Owner YUEYANG INTEGRATED TRADITIONAL CHINESE & WESTERN MEDICINE HOSPITAL SHANGHAI UNIV OF CHINESE TRADITIONAL MEDICINE
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