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Antisense nucleic acid sequence for SARS coronavirus gene and its pharmaceutical use

A technology of antisense nucleic acid and coronavirus, which is applied in the field of antisense nucleic acid sequence against SARS coronavirus gene and its medicinal application, and can solve the problem that the specificity is not very high

Inactive Publication Date: 2005-01-26
北京金赛狮反义核酸技术开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional drugs generally act on one or two sites on the spatial configuration of protein molecules with biological functions. Since the three-dimensional structure of proteins is very complex and there are few target sites that drugs can act on, the specificity will not be very high.

Method used

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  • Antisense nucleic acid sequence for SARS coronavirus gene and its pharmaceutical use
  • Antisense nucleic acid sequence for SARS coronavirus gene and its pharmaceutical use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Determination of the attack target of SA series antisense nucleic acid sequences and synthesis of antisense nucleic acid sequences.

[0043] Select the site of antisense nucleic acid attack on each gene segment of whole SARS coronavirus genetic map (about 30Kb).

[0044] The designed antisense nucleic acid was synthesized by the phosphoramidite solid-phase synthesis method using the Model 392 DNA-RNA synthesizer manufactured by APPLIEDBIOSYSTEMS and its supporting reagents, and then modified with thio. Synthetic raw materials are provided by synthesizer manufacturers, the main raw materials are: four diisopropyl β-cyanoethyl phosphoramidite monomers: adenosine (A), guanosine (G), thymidine (T), Cytosine (C); Four kinds of control pore glass powder (Control Pore Glass) solid-phase synthesis carrier (A, G, C, T); Activation reagent: tetrazole / acetonitrile capping reagent: acetic anhydride / dimethylpyrimidine / tetrahydrofuran, 1-methylpyrimidine / tetrahydrofuran;...

Embodiment 2

[0055] Embodiment 2: The chemical composition structure of SA series antisense nucleic acid sequence

[0056] Treat the synthesized and purified oligonucleotides with iodine to convert the phosphothioester bonds into phosphodiester bonds, and then use conventional DNA sequencing methods to sequence all antisense nucleic acid sequences, and the results show that the obtained antisense nucleic acids The sequences are consistent with the designed sequences; the molecular weights of all antisense nucleic acid sequences are determined by capillary electrophoresis, and found to be approximately equal to the theoretical molecular weights.

Embodiment 3

[0057] Embodiment 3: The inhibition of SA series antisense nucleic acid sequence for the African green monkey kidney cells infected with SARS:

[0058] 1. Verify with SARS virus-BJ-01 strain on the African green monkey kidney cell (Vero-E6) described in embodiment 3. Operated in P3 laboratory.

[0059] 2. The dose of SARS coronavirus is 10 -3 (Viral half infective dose TCID 50 =10 -7 , the virus dose used in this experiment is four orders of magnitude higher than the virus half infection dose), usually 2-3 days lesions.

[0060] 3. The dose of the drug is 1 μM

[0061] 4. Judging index (determination of efficacy by CPE method)

[0062] Cells without lesions:-

[0063] Cytopathy below 25%: +

[0064] Cytopathic in 26-50%: ++

[0065] Cytopathic in 51-75%: +++

[0066] Cytopathic at 76-100%: ++++

[0067] The calculation of cell viability was estimated according to the observation scale mentioned above. Cytopathy between + and +++ is calculated as the median of the cy...

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Abstract

The invention discloses an artificial synthesized antisense nucleic acid sequence targeting coronary virogene of Severe Acute Respiratory Syndrome (SARS), which is compatible with the nucleic acid sequence on specific positioning spots on the SARS coronavirus gene, thus can block the reproduction or expression in human body or animal body and achieve the goal of treating the disease. The invention also relates to the medicinal compositions containing this antisense nucleic acid sequence, and its pharmaceutical use.

Description

technical field [0001] The present invention relates to an antisense nucleic acid sequence against SARS coronavirus gene, a medicine containing the antisense nucleic acid sequence and its use. Background technique [0002] Severe Acute Respiratory Syndrome (hereinafter referred to as SARS), also known as atypical pneumonia, is an infectious disease caused by a new coronavirus. It first broke out in Guangdong, China, and then spread to neighboring countries and regions. As of By May 17, 2003, it had spread to 32 countries and regions, and more than 7,000 SARS cases had been reported. The onset is characterized by high fever, general malaise, headache, dry cough, dyspnea, hypoxemia, and a cavity in the lungs. Systemic interstitial infiltrates can be found. In severe cases, intubation or a ventilator is required to help the patient breathe, and death The rate is as high as about 15% (Kanchan Anand, John Ziebuhr, Parvesh Wadhwani and et al., Science, 2...

Claims

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Application Information

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IPC IPC(8): A61K31/7088A61P11/00A61P31/14C07H21/00C12N15/113
Inventor 王君刘健平
Owner 北京金赛狮反义核酸技术开发有限公司
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