Materials and methods for prevention and treatment of viral infections

A virus infection and compound technology, applied in the direction of antiviral agents, pharmaceutical formulations, plant raw materials, etc., can solve the problems of resistance mutation weakening the efficacy of antiviral drugs, and difficulty in treating compounds.

Active Publication Date: 2013-01-16
BAGI RES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

While these advances are impressive, the isolation and identification of useful therapeutic compounds from natural sources remains difficult and largely empirical
Also, as virus strains change over time, the emergence of resistance mutations further reduces the effectiveness of antivirals

Method used

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  • Materials and methods for prevention and treatment of viral infections
  • Materials and methods for prevention and treatment of viral infections
  • Materials and methods for prevention and treatment of viral infections

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0198] Preparation of Yinqiaosan (YQS) Extract

[0199] The YQS extract was prepared according to Chinese Pharmacopoeia (2005). Briefly, 3 g of mint and 2 g of nepeta were extracted twice with 20 times milli-Q water at reflux for 1 h. Volatile oil and water extracts were collected. The residue was combined with five other herbs including forsythia (5g), burdock seed (3g), tempeh (2.5g), bamboo leaves (2g) and licorice (2.5g), followed by 10-fold milli-Q The water was refluxed for 2 hours for extraction twice. Collect the supernatant and combine it with the previous aqueous extract. The resulting extract was then lyophilized under reduced pressure. The dried paste was combined with honeysuckle (5 g) and bellflower (3 g), and then extracted three times with 10 times MeOH. About 5 g of MeOH extract were obtained.

[0200] Fractionation of YQS Extracts for Antiviral Bioanalysis

[0201] The MeOH extract of YQS was fractionated by reverse phase HPLC (Lichrospher 100RPC18EC 5...

Embodiment 1

[0226] Example 1-Extraction and Identification of Forsythiaside A and Jacarcarone

[0227] A light yellow powder was obtained by repeatedly purifying the MeOH extract prepared from Forsythia using reverse-phase HPLC. The detailed process is summarized in Figure 1.

[0228] By bioactivity-guided purification using sequential HPLC, the antiviral molecule known as compound A1 eluted at approximately 6.8 min as a single compound (>95% pure) with UV absorbance maxima at 220, 290 and 330 (Fig. 2). Compound A1 13 The CNMR spectrum is shown in Figure 4, showing the following signals: δ 131.5 (C-1), 116.5 (C-2), 146.2 (C-3), 144.8 (C-4), 117.2 (C-5 ), 121.4(C-6), 72.4(C-α), 36.8(C-β), 104.6(C-1'), 75.3(C-2'), 76.0(C-3'), 72.5(C -4'), 74.9(C-5'), 67.8(C-6'), 102.4(C-1"), 72.1(C-2"), 72.4(C-3"), 74.1(C-3 ”), 70.0 (C-4”), 18.1 (C-5”), 127.8 (C-1”’), 115.2 (C-2”’), 147.0 (C-3”’), 149.9 (C- 4"'), 116.6 (C-5"'), 123.2 (C-6"'), 147.7 (C-7""), 114.8 (C-8"'), 168.4 (C-9"'). Compound A1 s...

Embodiment 2

[0230] Determination of the Cytotoxicity of Example 2-Forsythiaside A

[0231] After determining the inhibitory effect of forsythiaside A (compound A1) on influenza virus replication, the cytotoxicity of forsythiaside A (compound A1) was examined as follows. Briefly, Madin-Darby canine kidney (MDCK) cells were 5 The density of cells / well was seeded in a 24-well plate and incubated for 18 hours, then forsythiaside A (compound A1) was added at a concentration of 100ug / ml, or dimethyl sulfoxide (DMSO) was added in the control.

[0232] At 37°C, 5% CO 2 After 48 hours of incubation under conditioned conditions, cell viability was measured by the thiazolium blue tetrazolium bromide (MTT) assay. First, MTT was added to the cells at a final concentration of 0.1 mg / ml. After 90 minutes of incubation, the culture supernatant was removed and each well was treated with isopropanol for cell fixation. The MTT metabolite formazan was dissolved in isopropanol with constant shaking for 5 ...

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Abstract

The subject invention provides a novel and advantageous method for preventing and treating viral infection. Specifically exemplified herein are therapeutic uses of forsythoside A and jacaranone, compounds isolated from traditional Chinese medicinal material such as Fructus forsythiae (Lian Qiao). Also provided is use of a Yin Qiao San composition for preventing and treating viral infection.

Description

[0001] Cross-references to related applications [0002] This application claims the benefit of US Provisional Application No. 61 / 291,073, filed December 30, 2009, which is hereby incorporated by reference in its entirety. Background technique [0003] Viral infections cause many acute and chronic life-threatening diseases. An estimated 33.4 million people worldwide are infected with human immunodeficiency virus (HIV) (1) . In addition, an estimated 2 billion people are infected with hepatitis B virus and 600,000 people die each year from the acute or chronic consequences of the infection (2) . [0004] Influenza is an acute viral infection and one of the most widespread viral infections in the world. Major influenza A pandemics include the 1957 Asian influenza pandemic (H2N2), the 1968 Hong Kong influenza pandemic (H3N2), the H1N1 (Russian flu) re-emergence in 1970, the H5N1 in 1997 and 2003 Bird flu, and most recently the April 2009 swine flu (H1N1) outbreak. In Novemb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61K36/34A61K36/28A61P31/16A61K31/7032A61K31/215A61K31/192A61K36/899A61K36/634A61K36/538A61K36/534A61K36/484A61K36/48A61K36/35
CPCA61K36/534A61K36/33A61K36/484A61K36/48A61K31/7028A61K31/192A61K36/538A61K36/634A61K31/215A61K45/06A61K36/355A61P31/12A61P31/16Y02A50/30A61K2300/00
Inventor A·S-Y·刘L·H·C·杨A·H-Y·罗
Owner BAGI RES
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