Application of citrate ions and iron ions to inhibition of RNA viruses

A technology of RNA virus and citrate, applied in antiviral agents, medical preparations containing active ingredients, active ingredients of heavy metals, etc., can solve the problems of strong infectivity, fast transmission speed, and widespread epidemics

Active Publication Date: 2017-04-26
中国科学院上海免疫与感染研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Influenza viruses are prone to mutations, are highly contagious and spread quickly, and can easily cause large-scale epidemics

Method used

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  • Application of citrate ions and iron ions to inhibition of RNA viruses
  • Application of citrate ions and iron ions to inhibition of RNA viruses
  • Application of citrate ions and iron ions to inhibition of RNA viruses

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0081] Preparation of human primary DC cells

[0082] Peripheral blood mononuclear cells were obtained from fresh blood obtained from Shanghai Blood Center by Ficoll density gradient centrifugation. Monocytes were then enriched with CD14 magnetic beads. Mature DC cells were obtained by culturing in RPMI 1640 medium containing 40ng / mL GM-CSF and 20ng / mL IL-4 for one week.

[0083] Virus

[0084] EV71 virus FY573 strain (subgenotype C4a, GenBank access ion number: HM064456.1), HIV-1 recombinant virus, influenza virus PR8 strain, and human cytomegalovirus (HCMV) were all from Shanghai Pasteur Institute.

[0085] Real-time fluorescent quantitative PCR (Real-Time PCR)

[0086] Total RNA was extracted from RD or THP-1 cells using TRIzol reagent (Invitrogen), reversed into cDNA using a reverse transcription kit (Promega), and then detected using SYBR Green qPCR Master Mix (TOYOBO) and 7900HT Real-timePCR system (Applied Biosystems) Relative expression of genes. GAPDH gene was us...

Embodiment 1

[0108] The inhibitory effect of embodiment 1 ferric ammonium citrate (FAC) to EV71 virus

[0109] method:

[0110] 1.1 RD cells were infected with EV71 virus (MOI=0.1) and FAC (100 μM) was added at the same time. After 8 hours, the cytopathic changes were photographed; then RNA was extracted from the cells, and the viral RNA load was detected by Real-Time PCR; and by Western blotting (western blot) detection of the expression of EV71 protein VP1 in the cells;

[0111] 1.2 When EV71 infected RD cells, different doses (0, 10, 20, 50, 100 μM) of FAC were added. After 8 hours, Real-Time PCR was used to detect the viral RNA load in the cells, and the cell supernatant was tested for virus titer by TCID50 method.

[0112] 1.3 FAC (100 μM) was added while THP-1 cells were infected with EV71 virus (MOI=0.1), and the viral RNA load in the cells was detected by Real-Time PCR 12 hours later. And FAC (100μM) was added while THP-1 cells were infected with EV71 virus (MOI=0.1). After 24 ho...

Embodiment 2

[0115] The inhibitory effect of embodiment 2 ferric ammonium citrate (FAC) to HIV virus

[0116] Methods: DC cells differentiated from human primary peripheral blood mononuclear cells (PBMC) were used to infect HIV-1 (Gag(p24) 100ng / ml). After 8, 24, 48, and 72 hours, Real-Time PCR was used to detect virus Gag RNA load, mRNA expression of IFN-β, IL-1β and IL-6. FAC (100 μM).

[0117] Result: by figure 2 It can be seen that FAC can also inhibit the replication of HIV-1 in human primary DC cells ( figure 2 A). The secretion of cytokines caused by HIV virus infection is also not induced due to the inhibition of virus replication ( figure 2 B,C,D).

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Abstract

The invention provides an application of citrate ions and iron ions to inhibition of RNA viruses, particularly an application of an active ingredient to be used for preparing pharmaceutical composition for inhibiting the RNA viruses. The active ingredient at least comprises a substance A providing citrate ions C5H7O5COO<-> and a substance B providing iron ions in the molar ratio of the citrate ions to the iron ions being 1:8-8:1. The compounded active ingredient can effectively inhibit multiple RNA viruses, such as enterovirus, influenza virus, HIV and the like, when the citrate ions and the iron ions are applied in a specific proportion, the produced effect has certain correlation with the proportion. Besides, the substances providing the citrate ions and the iron ions are not limited, so that the compounded active ingredient is very easy to prepare, simple and cheap and has bright application prospect.

Description

technical field [0001] The invention relates to the field of biomedical treatment. Specifically, it relates to the application of specific contents of citrate ions and iron ions in inhibiting RNA viruses. Background technique [0002] As an infectious disease that seriously endangers children's health, hand, foot and mouth disease has become more prevalent in China in recent years. Since the Ministry of Health listed hand, foot and mouth disease as a Class C infectious disease under the Law on the Prevention and Control of Infectious Diseases for management, the number of cases of hand, foot and mouth disease in China has increased from 48,895 in 2008, with 126 deaths (accounting for 28% of Class C infectious diseases respectively). % and 67%), rising to 2,778,861 cases and 501 deaths in 2014 (accounting for 68% and 88% of Class C infectious diseases, respectively). [0003] HFMD is mainly caused by enterovirus 71 (EV71) and Coxsackievirus A16 (CoxsackievirusA16). The mai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K33/26A61K31/194A61P31/14A61P31/16A61P31/18
Inventor 孟广勋王洪斌
Owner 中国科学院上海免疫与感染研究所
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