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Method for predicting angiotonin II receptor agonist hypotensor function and use

A technology for angiotensin and antagonists, applied in the field of predicting the action of angiotensin II receptor antagonist antihypertensive drugs

Active Publication Date: 2011-06-15
SHENZHEN AUSA PHARM CO LTD +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the levels of folic acid and VB12 in the body are negatively correlated with tHcy, suggesting that supplementing folic acid and VB12 in hypertensive patients may delay the damage of their target organs

Method used

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  • Method for predicting angiotonin II receptor agonist hypotensor function and use
  • Method for predicting angiotonin II receptor agonist hypotensor function and use
  • Method for predicting angiotonin II receptor agonist hypotensor function and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Example 1: Determination of the Ala222Val polymorphism site of the MTHFR gene and prediction of the antihypertensive effect of the AT1 receptor antagonist antihypertensive drug nifedipine

[0082] (1) Determination of the genotype of the Ala222Val polymorphism site of the MTHFR gene:

[0083] (1) Extract the genomic DNA of the host cell:

[0084] (a) Add 30ml of erythrocyte lysate to the whole blood, shake slowly, and let stand at room temperature for 10 minutes. During this period, shake several times to completely lyse the erythrocytes;

[0085] (b) Centrifuge at 4°C at 2000 rpm for 10 minutes, remove the supernatant, break up the precipitated leukocytes on a rotary shaker, add 40ul of protease and 50ul of RNase, shake well, add 15ml of leukocyte lysate, Mix well in a 37°C water bath for 20 minutes, take it out, and put it in cold water;

[0086] (c) Add 4ml of cold protein precipitation solution, mix well and place in -20°C refrigerator for 5 minutes, take it out a...

Embodiment 2

[0118] Example 2: Determining the genotype of the MTRR gene Ile22Met (A145G) polymorphism and predicting the antihypertensive effect of the AT1 receptor antagonist antihypertensive drug nifedipine

[0119] (1) DNA extraction is the same as in Example 1, and the polymorphic site genotype determination method is similar to Example 1, wherein

[0120] The polymorphic site of the MTHFR Ile22Met gene and its flanking sequences were amplified with a PCR instrument. Corresponding to the "A (or Ile)" allele, the specific probe carries the VIC fluorescent reporter group; corresponding to the "G (or Met)" allele, the specific probe carries the FAM fluorescent reporter group.

[0121] Put the PCR plate that completed the PCR reaction into the 7900 fluorescent quantitative PCR instrument, and select the "AllelicDiscrimination" program to scan and judge the results:

[0122] The genotype of those who emit FAM fluorescence is Met / Met homozygote;

[0123] The genotype of those who emit VIC...

Embodiment 3

[0133] Embodiment 3: Measuring the Ala222Val polymorphic site kit composition (PCR-RFLP method) and application of MTHFR gene

[0134] 1. Kit components include:

[0135] Solution A: PCR buffer (PCR buffer), the components are KCL, Tris-HCl and MgCl2;

[0136] Solution B: deoxymononucleotide triphosphate (dNTP);

[0137] Liquid C: primer (Primer), synthesized by an oligonucleotide synthesizer; the sequence of the primer in the kit is:

[0138] Upstream primer: 5'-CAAAGGCCACCCCGAAGC-3', sequence listing 3 (SEQ ID No.3)

[0139] Downstream primer: 5'-AGGACGGTGCGGTGAGAGTG-3', sequence listing 4 (SEQ ID No.4)

[0140] Solution D: heat-resistant DNA polymerase (Taq); storage temperature is -20°C;

[0141] Solution E: restriction endonuclease HinfI; storage temperature is -20°C;

[0142] Solution F: restriction endonuclease buffer, the components are Tris-HCl, NaCl, MgCl2, dithiothreitol.

[0143] 2. The steps of using this kit to amplify functional polymorphic sites and their...

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Abstract

The present invention provides a kind of reagent kit including reagent for detecting the polymorphic site genotype of homocysteine metabolizing passage gene of nucleic acid template. By means of PCR-RFLP method or Taqman method, the detection is fast, safe, convenient, sensitive and reliable. The detection can predict the hypotensive effect of angiotonin II receptor 1 agonist hypotensor and predict the functional state of the target organ of the primary hypertension patient, so that the medicine may be selected based on the individual difference to raise clinical treating efficiency and safety and lower the toxic side effect risk and treating cost.

Description

technical field [0001] The invention relates to a method for predicting the action of angiotensin II receptor antagonist antihypertensive drugs and its application. By measuring the genotype of a specific site in a biological sample, the angiotensin II receptor 1 antagonist antihypertensive drugs are predicted drug effect. Background technique [0002] Hypertension is one of the most common chronic diseases and a major risk factor for stroke, coronary heart disease, diabetes, etc. Therefore, effective control of blood pressure has important clinical and public health significance. There are six types of drugs clinically used to treat hypertension, including: angiotensin II receptor (AT1-R) antagonists, β-blockers, angiotensin converting enzyme inhibitors (ACEI), calcium antagonists , α-adrenoceptor blockers and diuretics. But so far the control rate of hypertension is low, the data of our country and the United States show that the control rate is only 8.1% and 34% respect...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 张善春蒋善群王琳琳洪秀梅毛广运张岩李志平刘学邢厚恂唐根富王滨燕徐希平
Owner SHENZHEN AUSA PHARM CO LTD
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