CN112939928A

CN112939928A - Dendrobium officinale Kimura et Migo extract and its application

Info

Publication number: CN112939928A
Application number: CN202110107820.0A
Authority: CN
Inventors: 盛军; 字成庭; 王宣军; 袁文娟; 孙秀丽; 谢引荣; 杨豪男; 王丽霞; 张宁; 吴以龙; 施博雅; 王泽昊
Original assignee: Yunnan Agricultural University
Current assignee: Yunnan Agricultural University
Priority date: 2021-01-27
Filing date: 2021-01-27
Publication date: 2021-06-11

Abstract

The invention Discloses Dendrobium Candidins (DDCDs) and application of anti-inflammatory drugs thereof, wherein the structural formula of the compound is as follows:

Description

Dendrobium officinale Kimura et Migo extract and its application

Technical Field

The invention belongs to the technical field of cell biology, and particularly relates to dendrobium officinale Kimura et Migo extract and application thereof.

Background

Inflammation is a defense reaction of living tissues having a vascular system against an injury factor, and is a defense reaction of the body against a stimulus. Inflammation can be generally divided into acute inflammation and chronic inflammation, with acute inflammation manifested by redness, swelling, heat, pain, and dysfunction; chronic inflammation is clearly harmful to the body and may lead to cancer, diabetes, pulmonary diseases, diseases of the cardiovascular and nervous systems, etc.

The research proves that the dendrobium officinale fermented polysaccharide has an inhibiting effect on mouse ulcerative colitis; the dendrobium nobile lindl alkaloid has the protection effect on the BV2 cell inflammatory reaction induced by lipopolysaccharide; the dendrobium candidum glycoprotein can promote wound healing by regulating cell inflammatory response and is related to NF-kB signal channel activation; stilbene compounds in herba Dendrobii have effects of inhibiting various tumor cells. The dendrobium candidum element is a characteristic component of dendrobium candidum and is a bibenzyl derivative. At present, few research literatures are available for dendrobium officinale Kimura et Migo, and it is necessary to research the biological activity of dendrobium officinale Kimura et Migo so as to obtain the effective effect of dendrobium officinale Kimura et Migo on inflammation.

Factors associated with inflammation include: nitric Oxide (NO), interleukin-alpha (IL-alpha), interleukin-beta (IL-beta), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), and the like. Various diseases caused by chronic inflammation can be treated by inhibiting inflammatory factors. Therefore, inflammatory factor inhibitory drugs are hot spots for the development of new anti-inflammatory drugs at present.

Disclosure of Invention

The first purpose of the invention is to provide a dendrobium officinale Kimura et Migo extract.

The second purpose of the invention is to provide the application of the dendrobium candidum in preparing the medicines for inhibiting the release of IL-6, MMP3 or TNF-alpha inflammatory factors.

The invention achieves the purpose of the first invention, and adopts the technical scheme that: the dendrobium officinale extract has the following structure:

the compound is named as dendrobium officinale Kimura et Migo (Dendrocandins, DDCDs).

The dendrobium officinale element structurally comprises:

R₁position H, OH or OCH₃；

R₂Position H, OH or OCH₃；

R₃Position H, OH or OCH₃；

R₄Position H, OH or OCH₃；

R₅Position H, OH or OCH₃；

R₆Position H, OH or OCH₃；

R₇Position H, OH or OCH₃。

The invention achieves the second invention purpose, and the adopted technical scheme is as follows:

the dendrobium candidum extract is applied to the preparation of anti-inflammatory drugs, precursors of the anti-inflammatory drugs or lead compounds of the anti-inflammatory drugs.

The medicine for inhibiting the release of IL-6, MMP3 or TNF-alpha inflammatory factor contains dendrobium officinale Kimura et Migo with a therapeutically effective dose and a pharmaceutically acceptable carrier.

The above-mentioned drugs may be in a form suitable for oral use, and the dosage form may be a tablet, troche, lozenge, aqueous or oily suspension, dispersible powder or granule, emulsion, liquid, hard or soft capsule, or sugar-filled or brewed preparation.

The invention has the beneficial effects that:

the invention provides dendrobium candidum with a general structure, which has good effect of inhibiting IL-6, MMP3 or TNF-alpha inflammatory factor release, especially has good anti-inflammatory activity, and provides an important experimental basis for further preventing inflammation.

Drawings

FIG. 1 is a structural formula of Dendrobium officinale Kimura et Migo;

FIG. 2 is a graph showing the results of the effect of Dendrobiicin and LPS on cells;

FIG. 3 is a graph showing the change of Dendrobium officinale Kimura et Migo in the inflammatory factor IL-6 induced by LPS;

FIG. 4 is a graph showing the expression of inflammation-associated proteins (MMP 3, TNF-. alpha.) in inflammation induced by Dendrobium officinale Kimura et Migo LPS.

Detailed Description

In order that the objects, technical solutions and advantages of the present invention will be more apparent, preferred embodiments of the present invention will be described in detail below to facilitate understanding of the skilled person.

The dendrobium officinale extract has the following structure:

R₁position H, OH or OCH₃；

R₂Position H, OH or OCH₃；

R₃Position H, OH or OCH₃；

R₄Position H, OH or OCH₃；

R₅Position H, OH or OCH₃；

R₆Position H, OH or OCH₃；

R₇Position H, OH or OCH₃。

Specifically provides three dendrobium candidum elements:

the Dendrobium candidum extract can be obtained by extracting and separating Dendrobium candidum or other plants containing the similar compounds by a chromatographic separation method, or artificially synthesizing. The dendrobium officinale extract is preferably as follows:

(1) dendrobium officinale Kimura et Migo W (DCDD W)

The molecular formula is as follows: c₂₅H₂₆O₆

Molecular weight: 422

Shape: white amorphous powder

Plectrum bopp data:

¹H-NMR (500 MHz, CD₃OD) δ 7.18–7.15 (m, 5H, ArH), 7.10–7.07 (m, 1H, ArH), 6.97–6.95 (m, 2H, ArH), 6.46 (s, 2H, C^2'-H, C^6'-H), 6.05 (m, 1H, C^7''-H), 4.23 (m, 1H, C^8''-H), 3.79 (s, 6H, 2 × OCH₃), 3.31 (s, 3H, OCH₃), 2.36–2.34 (m, 2H), 2.01–2.00 (m, 2H); ¹³C-NMR (125 MHz, CD₃OD,) δ 149.1, 142.6, 141.6, 135.7, 134.0, 129.4, 129.3, 139.2, 129.1, 126.9, 123.7, 106.8, 98.9 (C-8^''), 74.5 (C-7^''), 60.2 (C-9^''), 56.9 (OCH₃), 38.4, 33.7 MS-ESI m/z (%): 421 ([M – H]^–, 100)。

(2) dendrobium officinale X (DCDD X)

The molecular formula is as follows: c₂₅H₂₆O₇

Molecular weight: 438

Shape: white amorphous powder

Plectrum bopp data:

¹H-NMR (500 MHz, CD₃OD) δ 7.02 (d, 2H, J = 8.25 Hz, C^2'-H, C^6'-H), 6.84 (d, 1H, J = 8.35 Hz, C⁵-H), 6.71 (m, 4H, C²-H, C⁶-H, C^3'-H, C^5'-H,), 6.31 (s, 2H, C^2''-H, C^6''-H), 4.29 (d, 1H, J = 2.55 Hz, C^8''-H), 3.91 (d, 2H, J = 5.10 Hz, C^9''-CH₂), 3.85 (m, 6H, C^3''-OCH₃, C^5''-OCH₃), 3.40 (d, 1H, J = 5.40 Hz, CH₂OH), 2.76 (s, 4H, C^α-CH₂, C^α'-CH₂); ¹³C-NMR (CDCl₃, 125 MHz) δ 153.8 (C-4^'), 146.9 (C-3^'', C-5^''), 140.7 (C-3), 133.9 (C-1^', C-4^''), 132.8 (C-1), 129.3 (C-2^', C-6^'), 122.2 (C-6), 119.4 (C-5), 115.3 (C-3^', C-5^'), 104.9 (C-2^'', C-6^''), 56.3 (C^3''-OCH₃, C^5''-OCH₃), 37.4 (C-α, C-α^'); MS-ESI m/z (%): 437 ([M - H]^-, 100).

(3) dendrobium officinale Kimura et Migo Y (DCDD Y)

The molecular formula is as follows: c₂₆H₂₈O₆

Molecular weight: 452

Shape: white amorphous powder

Plectrum bopp data:

¹H-NMR (500 MHz, CD₃OD) δ 6.90 (d, 2H, J = 8.50 Hz, C^2'-H, C^6'-H), 6.76 (d, 2H, J = 8.50 Hz, C^3'-H, C^5'-H), 6.34 (s, 2H, C^2''-H, C^6''-H), 5.43 (s, 1H, C^7''-H), 4.30 (d, 1H, J = 2.58 Hz, C^8''-H), 3.89 (d, 2H, J = 3.80 Hz, C^9''-CH₂), 3.86 (m, 9H, C^4'-OCH₃, C^3''-OCH₃, C^5''-OCH₃), 3.40 (d, 1H, J = 5.42 Hz, CH₂OH), 2.76 (s, 4H, C^α-CH₂, C^α'-CH₂); ¹³C-NMR (CDCl₃, 125 MHz) δ 157.9 (C-4^'), 146.9 (C-3^'', C-5^''), 140.7 (C-3), 133.9 (C-1^', C-4^''), 132.7 (C-1), 129.0 (C-2^', C-6^'), 122.1 (C-6), 113.8 (C-3^', C-5^'), 104.8 (C-2^'', C-6^''), 56.4 (C^3''-OCH₃, C^5''-OCH₃), 55.3 (C^4'-OCH₃), 37.4 (C-α, C-α^'); MS-ESI m/z (%): 451 ([M - H]^-, 100).

example evaluation of anti-inflammatory Effect of Dendrobii officinalis

This example uses Raw264.7 macrophages with compounds lipopolysaccharide LPS (induced inflammation), dexamethasone (positive control) and Dendrobium officinale X (DDCD) (experimental group).

The anti-inflammatory effect test steps of the dendrobium officinale Kimura et Migo extract are as follows:

1) inoculating Raw264.7 cells onto a 96-well plate, changing into a serum-free culture medium after the cells adhere to the wall, treating with dexamethasone and dendrobium officinale essence, adding LPS after 1 hour, culturing for 24 hours, adding MTT, and detecting the activity of the cells;

2) raw264.7 cells were seeded 2X 10⁵Putting the cells on a 24-well plate, adding dexamethasone and dendrobium officinale pigment for treatment after the cells adhere to the wall, adding LPS after 1 hour, culturing for 24 hours, taking cell supernatant, and detecting the expression content of IL-6 inflammatory factors by an ELISA kit;

3) will be 3X 10⁶The Raw264.7 cells are inoculated on a small dish, dexamethasone and dendrobium officinale are used for processing after the cells are attached to the wall, LPS is added after 1 hour, protein in the cells is extracted after 24 hours of culture, and the expression conditions of cell inflammation related proteins such as MMP3, TNF-alpha and the like are detected by using a western blotting experimental method.

The anti-inflammatory effect of the dendrobium officinale Kimura et Migo is shown in figures 2-4.

Compared with an LPS treatment group, after treatment by dexamethasone and dendrobium officinale Kimuramic acid with different concentrations, the cell activity is obviously increased along with the increase of the concentration of the dendrobium officinale Kimuramic acid;

the ELISA kit experiment result shows that compared with an LPS (low-cholesterol) single treatment group, the content of an inflammatory factor IL-6 in cells of the LPS single treatment group is remarkably reduced along with the increase of the concentration of the dendrobium officinale pigment after the treatment by dexamethasone and the dendrobium officinale pigment with different concentrations;

the results of protein immunoblotting experiments show that compared with LPS treatment groups, the content of inflammation-related proteins (MMP 3 and TNF-alpha) in the extracted cell proteins is remarkably reduced after treatment with dendrobium officinale Kimura et Migo with different concentrations.

Experiments show that the dendrobium officinale Kimura et Migo extract has a good inhibition effect on inflammation induced by LPS, and provides an important experimental basis for further preventing inflammation by the dendrobium officinale Kimura et Migo extract.

Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the scope of the invention as defined by the appended claims.

Claims

1. Dendrobium officinale Kimura et Migo extract, which is characterized in that: has the following structure:

2. The dendrobium officinale Kimura et Migo of claim 1, wherein: the structure includes:

R₁position H, OH or OCH₃；

R₂Position H, OH or OCH₃；

R₃Position H, OH or OCH₃；

R₄Position H, OH or OCH₃；

R₅Position H, OH or OCH₃；

R₆Position H, OH or OCH₃；

R₇Position H, OH or OCH₃。

3. The use of the dendrobium officinale Kimura et Migo of claim 1 or 2 in the preparation of a medicament for inhibiting the release of IL-6, MMP3 or TNF- α inflammatory factor.

4. Use according to claim 3, characterized in that: the medicine contains the dendrobium officinale Kimura et Migo with effective treatment amount and a pharmaceutically acceptable carrier.

5. Use according to claim 3 or 4, characterized in that: the dosage form of the medicament can be tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, solutions, hard or soft capsules, or sugar or brewing agents.