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Compound and application thereof to detection of mycobacterium tuberculosis

A technology of compounds and derivatives, applied in the field of detection of Mycobacterium tuberculosis, can solve problems that need to be developed

Active Publication Date: 2017-10-20
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, current TB diagnostic methods and even M. tuberculosis detection techniques are still under development

Method used

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  • Compound and application thereof to detection of mycobacterium tuberculosis
  • Compound and application thereof to detection of mycobacterium tuberculosis
  • Compound and application thereof to detection of mycobacterium tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Fluorescence response of compound shown in embodiment 1 formula I to Rv2466c enzyme

[0107] Instrument: multifunctional microplate reader ( Flash, Thermo Scientific)

[0108] Reagents and materials: compound (I-1 to I-28, I-69), Rv2466c, MSH, Tris-Cl (pH 7.5), DMSO, 96-well microtiter plate (Corning, 3603)

[0109] Operation: 1) Prepare 50mM Tris-Cl (pH 7.5) buffer solution containing 0.5mM MSH, and put 20mL buffer solution into two sterile tubes. 2) Add a certain amount of Rv2466c to one of the tubes so that the final concentration is 200 μg / mL, which is the enzyme-containing working solution, and add the same amount of Tris-Cl buffer solution to the other tube as the enzyme-free control solution. 3) The compounds of the examples were respectively dissolved in DMSO to prepare a solution of 25 μg / mL, and 20 μL of each compound was added to four wells of a 96-well plate, two of which were used as a control group without enzymes, The other two wells are the enzyme-c...

Embodiment 2

[0115] On the basis of Example 1, the inventor followed the steps below, taking compound I-6 as an example to study the fluorescence characteristics of compound I of the present invention and its nitro reduction product:

[0116] According to the results of mass spectrometry, compound I-6 is reduced to I-6a, I-6b and I-6d in the presence of Mtb, and intermediate I-6c is inferred from the structure of I-6d. Therefore, it is speculated that I -6 has the following reduction and hydrolysis processes:

[0117]

[0118] Then proceed as follows:

[0119] Instrument: multifunctional microplate reader ( Flash, Thermo Scientific)

[0120] Reagents and materials: Compounds (I-6, I-6a, I-6b, I-6c, I-6d), DMSO, 7H9 medium, 96-well enzyme plate (Corning, 3603)

[0121] Operation: 1) Take an appropriate amount of compound I-6, I-6a, I-6b, I-6c, and I-6d powder, and prepare it into a 10 mg / mL DMSO solution. 2) Dilute the DMSO solution 1000 times with 7H9 medium to obtain a test solut...

Embodiment 3

[0125] On the basis of Example 1-2, the inventor followed the steps below, taking compound I-6 as an example, to study the fluorescence response of compound I of the present invention to Rv2466c enzyme:

[0126] Instrument: multifunctional microplate reader ( Flash, Thermo Scientific)

[0127] Reagents and materials: compound I-6, Rv2466c, DTT (dithiothreitol), MSH, Tris-Cl (pH 7.5), DMSO, 96-well microtiter plate (Corning, 3603)

[0128] Operation: Steps 1)-4) are the same as in Example 2. In step 5), it is necessary to track and detect the fluorescence value under excitation at 390nm and emission at 470nm. Wherein, the reaction conditions are: 0.5 mM MSH, 1 mM DTT, 200 μg / mL Rv2466c and 2.5 μg / mL Example compound I-6.

[0129] see results Figure 4 .

[0130] Figure 4 In the presence of DTT and MSH, after Rv2466c catalyzed the reduction of compound I-6, the detection results were tracked under the conditions of excitation light and emission light (390 / 470nm). Thus, ...

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Abstract

The invention provides a compound and an application thereof to detection of mycobacterium tuberculosis. The compound is a compound or derivative thereof shown in the formula I as in the description. The compound or derivative thereof shown in the formula I is taken as a detection reagent and contacts a to-be-detected sample. Based on fluorescence change of the to-be-detected sample before or after the contact, whether mycobacterium tuberculosis exists in the to-be-detected sample can be effectively determined, and the result is accurate and reliable. Further, the detection information can be effectively used for diagnosing whether a suspected patient, where the to-be-detected sample comes, suffers from diseases related to mycobacterium tuberculosis. Therefore, tuberculosis positive patients (active tuberculosis patients) and mycobacterium tuberculosis carriers can be detected.

Description

technical field [0001] The invention relates to the technical field of mycobacterium tuberculosis detection, in particular to a compound and its application in the detection of mycobacterium tuberculosis. Background technique [0002] In 1882, German scientist Robert Koch first proved that Mycobacterium tuberculosis (Mtb) is the pathogenic bacteria of tuberculosis. Mycobacterium tuberculosis, referred to as Mycobacterium tuberculosis, is the most common infection in the lungs of humans. Common symptoms of a lung infection include chest pain and a protracted productive cough. According to the World Health Organization (WHO) estimate, in 2015, there were about 10.4 million new tuberculosis cases in the world, of which 1.2 million patients were co-infected with AIDS, and 1.4 million people died of tuberculosis. About 4.6% of the newly discovered cases are multidrug-resistant tuberculosis (MDR-TB) infection, and 105 countries have confirmed cases of extensively drug-resistant ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D493/04C09K11/06G01N21/64
CPCC07D493/04C09K11/06C09K2211/1029C09K2211/1033C09K2211/1088G01N21/6428G01N2021/6432
Inventor 刘刚穆然
Owner TSINGHUA UNIV
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