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Scutellaria barbata extract as well as preparation method and application thereof

A technology for extracts and medicines of Scutellaria barbata, which is applied in the directions of pharmaceutical formulations, plant raw materials, medical preparations containing active ingredients, etc. Replication activity and other issues, to achieve a good inhibitory effect, the effect of great application prospects

Inactive Publication Date: 2015-05-27
广州市爱菩新医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the anti-EBV virus cleavage and replication activity of Scutellaria barbata extract and its neocrotane diterpene components in the prior art

Method used

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  • Scutellaria barbata extract as well as preparation method and application thereof
  • Scutellaria barbata extract as well as preparation method and application thereof
  • Scutellaria barbata extract as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The method for preparing compound 1-4 from barbata (Scutellaria barbata):

[0031] Scutellaria barbata (Scutellaria barbata) whole plant (3.5Kg), dried in the sun, pulverized, leached 3 times with 95% ethanol at room temperature, each time for 1 day, combined the concentrated solutions to obtain crude extract (362g), respectively washed with petroleum ether , ethyl acetate, n-butanol extraction, ethyl acetate layer (183g) directly through macroporous resin (D101), with ethanol / water (0:100, 10:90, 20:80, 30:70, 40:60 , 50:50, 60:40, 70:30, 80:20:, 95:5, 100:0, V / V) gradient elution, one part was collected for each 450mL, and the same part was combined for TLC detection to obtain a total of six Fraction Fr.1-6. Parts Fr.3-5 were subjected to silica gel column chromatography (200-300 mesh), Sephadex LH-20, C 18 Column and semi-preparative liquid chromatography column, repeated separation, the following 4 compounds were obtained.

[0032] Compound 1 (new compound):

[...

Embodiment 2

[0049] Embodiment 2 Pharmacodynamic related experiments of the 4 compounds obtained in Example 1

[0050] Determination of inhibitory activity of compounds 1-4 of the present invention on EBV cleavage and replication.

[0051] (1) Cell culture: in vitro culture of P3HR-1 cells (primary exudative lymphoma cell line, containing EBV in latent infection stage), using 10% fetal bovine serum, streptomycin (100 μg / ml), The RPMI1640 medium of penicillin (100 units / mL) was maintained at 37° C. under the condition of 5% carbon dioxide concentration and subcultured.

[0052] (2) Drug intervention: adjust the density of P3HR-1 cells in the logarithmic growth phase to 3×10 5 cells / mL, use 20ng / mL of 12-O-tetradecanoylphorbol-13-ethyl ester (TPA) and sodium butyrate (0.3mM) to induce P3HR-1 cells to enter the lytic replication phase. DMSO was used to prepare drug solutions with different concentrations (150, 100, 50, 20, 10, 1, 0.1, 0 μM) of the compounds 1-4 to be tested. After P3HR-1 c...

Embodiment 3

[0059] Example 3 Compound 1-4 of the present invention is tested for host cell toxicity

[0060] (1) Cell culture: P3HR-1 cells (primary exudative lymphoma cell line, containing EBV in latent infection stage) were cultured in vitro. The RPMI1640 medium containing 10% fetal bovine serum, 400ug / ml G418, and 100ng / ml doxycycline was used for routine maintenance and passage at 37°C and 5% carbon dioxide concentration.

[0061] (2) Test method: adjust the density of P3HR-1 cells in the logarithmic growth phase to 3×10 5 cells / ml, the cells were treated with different concentrations of compound 1-4 (150, 100, 50, 20, 10, 1, 0.1, 0 μM), and three parallel wells were set up for each concentration. After 2 days, trypan blue staining was used, and light Count the number of viable cells under a microscope.

[0062] (3) Result processing: according to the formula: relative toxicity = 1-livecell compound+ / live cell compound_ Calculate the relative toxicity and median lethal dose (CC) ...

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Abstract

The invention belongs to the technical field of Chinese herbal medicine extraction and in particular discloses a scutellaria barbata extract as well as a preparation method and application thereof. Two compounds in a new structure and two compounds in a known structure are separated from the scutellaria barbata extract. The four compounds have good inhibiting effects on EB (Epstein-Barr) virus replication and have low toxic and side effects; therefore the four compounds have great application prospects in preparing medicines for preventing EB virus infection.

Description

technical field [0001] The invention relates to the technical field of extraction of traditional Chinese medicinal materials, and more specifically relates to an extract of Scutellaria barbata and its preparation method and application. Background technique [0002] Human herpesviruses can be divided into α, β and γ subtypes according to their biological characteristics, among which Epstein-Barr virus (Epstein-Barr virus) belongs to γ ​​subtype, also known as type 4 human herpesvirus (HHV-4). Epstein-Barr virus was first discovered by Epstein and Barr in 1964 from a lymphoma cell line cultured in vitro from African children's malignant lymphoma (Burkitt lymphoma), and a new herpes virus was discovered by electron microscopy, and it was named Epstein-Barr virus. The genome of EBV is a linear double-stranded DNA with a length of 172kb and encodes about 100 genes. Like human herpesviruses, EBV has two life cycles, a latent and lytic phase. In the latent phase, EBV expresses o...

Claims

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Application Information

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IPC IPC(8): A61K36/539C07D493/10A61K31/365A61P31/22
Inventor 顾琼吴泰宗徐峻袁岩王倩王彦
Owner 广州市爱菩新医药科技有限公司
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