JP2011162462A - Extract of antrodia cinnamomea fruit body and usage thereof, usage of antrocamphin a and method of inhibiting formation of inflammation inductible molecule - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide anti-inflammatory usage of Antrodia cinnamomea fruit body derivatives. <P>SOLUTION: The Antrodia cinnamomea fruit body derivatives include an extract from Antrodia cinnamomea fruit body with ethanol, an extract from Antrodia cinnamomea fruit body with anhydrous propionic acid Antrodia cinnamomea fruit body, and Antrocamphin A. The ethanol extract, the anhydrous propionic acid extract, and the Antrocamphin A have an effect of reducing the formation of nitrogen monoxide synthetic enzyme or cyclooxygenase 2 to inhibit the formation of an inflammation inducible molecule. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

TECHNICAL FIELD The present invention relates to a derivative of Benixunotake mushroom fruit body, and more particularly, to an anti-inflammatory use of the anti-inflammatory usage of the body of Benicus nokitake fruit body.

Inflammatory reaction is one of the main features in many pathological conditions, but lipopolysaccharide (LPS, also known as endotoxin) on the surface of bacteria activates macrophages in the host body, taking bacterial infection as an example. However, in the normal state, in the activation process, the entire immune system of the host is activated and the purpose of eliminating the pathogen is achieved. However, in some abnormal physiological reactions (those that have occurred or are a major cause of disease development), activated macrophages cause the production of excessive amounts of nitric oxide (NO), In many pathological conditions, control over the inflammatory response may be an important part of disease treatment because it will accelerate host cell death.

Tumor necrosis factor-α (TNF-α), known as inflammatory stimuli, nitric oxide (NO) and prostatin E ₂ (prostaglandin-E ₂ , PGE ₂ is the metabolism of arachidonic acid in the cyclooxygenase (COX) route) It is an important part of the immune response, for example, the development of sepsis or septic shock, rheumatoid arthritis and arthrosclerosis A lot of things were detected. Therefore, the development of new drugs suppresses the genes for nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2), and inhibits the main receptor or enzyme activity of signal transduction induced by lipopolysaccharide (LPS). , emphasis is placed on reducing the nitrogen monoxide (NO) generation of prostate element E _2.

Antrodia cinnamomea is a kind of fungus and is seen as a very rare Chinese herbal medicine, and many scientific research results show that methyl alcohol extract and hot water extract from Benix noctum mycelium, The fruit body methyl alcohol extract, ethanol extract and propionic anhydride extract were found to have excellent anti-cancer activity. And some studies pointed out that the mycelium and fruiting bodies of Benix nokitake are compounds with anti-inflammatory potential that contain some triterpenoid compounds. However, it is necessary to further elucidate the mechanism of action and physiological activity of Benix octopus and its included triterpenoid compounds that control the inflammatory response.

The object of the present invention is to provide a derivative of Benicus nokitake fruiting body and its use, thereby suppressing the production of pro-inflammatory molecules and having an excellent anti-inflammatory function.

In order to achieve the above-mentioned object, the present invention provides a fruit body of a dried venomus mushroom, which is obtained by a procedure of extracting the fruit body of the dried venus mushroom with ethanol at a constant temperature. It provides an extract of the fruiting body of Benicus mushroom.

The present invention also provides a fruit body of the dried Benix octopus, which is obtained by extracting ethanol from the fruit body of the dried Benix octopus at a constant temperature. To obtain a concentrated product, and to obtain an extract of the fruit body of Benicus edodes as the propionic anhydride extract obtained by partitioning the concentrated product with propionic anhydride and water. It is characterized by.

The drying method is performed by freeze drying.
The temperature is 42 to 45 ° C.
The ethanol is characterized by being made of ethanol with a volume percentage of 95%.

The concentration method is performed by vacuum concentration.
The propionic anhydride is characterized by being carried out with 50% volume percent propionic anhydride.

The present invention further provides the use of the extract characterized by suppressing the production of pro-inflammatory molecules.
The extract is characterized by inhibiting the pro-inflammatory molecule, nitric oxide (NO) or prostate element E ₂ (PGE ₂ ).
The extract is characterized by reducing nitric oxide synthase and cyclooxygenase 2 to suppress the production of the nitric oxide (NO) and the prostate element E ₂ (PGE ₂ ), respectively.

The present invention still further provides the use of antrocamfin A, which is characterized by suppressing the production of nitric oxide (NO).
The antrocampin A is characterized in that it reduces nitric oxide synthase and suppresses the production of the nitric oxide (NO).
In addition, the present invention provides an application of antrocamphin A that is characterized by inhibiting the production of prostate element E ₂ (PGE ₂ ).
The antrocampin A is characterized in that it reduces cyclooxygenase 2 and suppresses the production of the prostate element E ₂ (PGE ₂ ).

The present invention also provides a method for suppressing pro-inflammatory molecule production, comprising contacting a subject with any one of the extracts according to claim 17 or antrocampin A.
The pro-inflammatory molecule is characterized by being nitric oxide (NO) or prostate element E ₂ (PGE ₂ ).

In summary, in the present invention, the anti-inflammatory effect of the derivative of Benix nokitake fruit body in the present invention is further clarified in the physiological activity and action mechanism of the anti-inflammation caused by the derivative of Benix nokitake fruit body presented in the present invention. Became.

The graph which shows the suppression rate of nitric oxide production | generation which the ethanol extract of this invention, a propionic anhydride extract, and a water-soluble fraction have. The graph which shows the production amount of nitric oxide synthase (iNOS) which the ethanol extract of this invention influenced. The graph which shows the production amount of cyclooxygenase 2 (COX-2) which the ethanol extract of this invention influenced. The graph which shows the suppression rate of a nitric oxide production | generation which 17 fractions of the propionic anhydride extract of this invention have. The graph which shows the production amount of the messenger ribonucleic acid (mRNA) of nitric oxide synthase (iNOS) which an antrocamphin A influenced. The graph which shows the production amount of the messenger ribonucleic acid (mRNA) of cyclooxygenase 2 (COX-2) which the antrocampin A influenced. The graph which shows the production amount of nitric oxide synthase (iNOS) which antrocamphin A influenced. The graph which shows the production amount of cyclooxygenase 2 (COX-2) which antrocamphin A influenced.

The present invention relates to a derivative of Benicaria mushroom fruit body and its use, thereby suppressing the production of pro-inflammatory molecules and having an excellent anti-inflammatory function. The “derivative” means an extract of Benix octopus fruit body and its component compounds, and more specifically, “derivative” as used in the present invention means an ethanol extract of Benix octopus fruit body, Propionic anhydride extract or antrocampin A.

Lipopolysaccharide (LPS) is composed of lipids and polysaccharides, and is a major molecule on the cell surface of Gram-negative bacteria, also called endotoxin. Lipopolysaccharide causes a severe inflammatory response in the host body, but this severe inflammatory response is usually an immune response activated by macrophages, and therefore the present invention depends on the mouse body or macrophage strain induced by lipopolysaccharide. Experimenting with an inflammatory reaction serves as a model for examining the role of the fruit body of Benix nokitake in the present invention in relation to an anti-inflammatory reaction.

The following examples are given for the purpose of illustrating the present invention in detail and should not limit the scope of the claims of the present invention.

Experimental design [Material]
The fruit body of Benicus mushroom used in the present invention is a wild strain collected in Kaohsiung, Taiwan, and fetal bovine serum (FBS) is purchased from Gibco BRL (Invitrogen, Grand Island, NY), dimethyl sulfoxide (DMSO) , Penicillin, lipopolysaccharide, Escherichia coli 0127: 138 / LPS) MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolebromide (Grease) and Grease All were purchased from Sigma-Aldrich (St Louis, MO). All chemicals and solvents used in the present invention are of reagent or HPLC class.

[Experimental mouse]
The experimental mouse was a four-week-old male ICR mouse (about 25-28 g) purchased from BioLasco (Taipei, Taiwan). Before the experiment, the temperature was 25 ± 2 ° C. and the relative humidity was used to acclimate the experimental mouse to the environment. 55 ± 5%, light / dark cycle is 12 hours (more specifically, from 6 am to 6 pm every day), and is kept for at least one week under conditions where food and water are freely available I left it. All animal experimentation procedures were conducted in accordance with the provisions for laboratory animal care management (Guide for the Care and Use of Laboratory Animals), usage guides and animal protection laws in Taiwan, and after obtaining approval from local moral committee members.

[Cell culture]
In the present invention, the production amounts of nitric oxide and prostatic element E ₂ (PGE ₂ ) were examined in the mouse macrophage strain RAW 267.4. In the culture of RAW267.4 cells, first, RAW267.4 cells cultured in the culture medium (75 cm ² ) were inoculated into a 96-well culture plate at an inoculation density of 2 × 10 ⁵ cells / well, and the culture box was used. (37 ° C., 5% CO ₂ ), and the culture medium was obtained from ATCC (American Type Culture Collection), using the culture medium DMEM, and fetal bovine serum (FBS) 10% And penicillin (100 units / ml) and streptomycin (100 g / ml) are cultured.

[Western blot analysis]
After obtaining the total protein of the cell, a similar amount of protein is added to the 5-7% gradient electrophoresis gel pore (SDS-PAGE) and resolved at 300 mA for 90 minutes. After the protein blotting (100 V, 1 hour) separated by the above was transferred to a polyvinylidene fluoride membrane (PVDF, Immobilon, Millipore, Bedford, MA), the polyvinylidene fluoride membrane was blocked with a blocking buffer (blocking) buffer, 10% w / v skim milk powder in TBST buffer) and act for 1 hour.

Then, the polyvinylidene fluoride membrane is applied to an anti-nitrogen oxide synthase antibody solution (anti-iNOS, 1: 1000, Cayman Chemicals) or an anti-cyclooxygenase 2 antibody solution (anti-COX-2, 1: 1000, Cayman Chemicals). And then washed twice with TBST buffer 0.1% (TBS buffer containing 0.1% Tween 20) to wash off non-specifically bound antibody antigens.
Then, after immersing the polyvinylidene fluoride membrane in an anti-rabbit secondary antibody solution of horseradish peroxidase, chemiluminescent reagent (enhanced chemiluminescene reagents, ECL, PCL) was added. The fluorescence intensity of each set is detected, and the amount of β-actin produced is a protein control.

[Reverse transcription polymerase chain reaction]
Extract total ribonucleic acid (total RNA) from extracted cells with Trizol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA), and transfer messenger ribonucleic acid (mRNA) of iNOS, COX-2 sum G3PDH by reverse transcription polymerase chain reaction Reverse transcription into cDNA and then measuring production of iNOS, COX-2 and G3PDH by quantitative real-time polymerase chain reaction (real time-PCR, Applied Biosystems).

The quantitative real-time polymerase chain reaction detects a PCR product with a DNA-binding fluorescent agent Green Nucleic Acid Gel Stain (SYBR Green), and the temperature cycle is 95 ° C. for 5 minutes after one cycle, and 95 cycles. 1 minute at ° C, 45 seconds at 55 ° C, and 30 seconds at 72 ° C for 40 cycles, but the primer sequence is Inos forward 5'-TCC TAC ACC ACA CCA AAC-3 '; iNOS reverse 5'-CTC CAA TCT CTG CCT ATC C-3 ′; COX-2 forward 5′-CCT CTG CGA TGC TCT TCC-3 ′; COX-2 reverse 5′-TCA CAC TTA TAC TGG TCA AAT CC-3 ′; G3PDH Orientation 5'-TCA ACG GCA CAG TCA AG -3 ′; G3PDH reverse 5′-ACT CCA CGA CAT ACT CAG C-3 ′, G3PDH is a housekeeping gene, and has a stable and large amount of gene in cells. Standardize the amount of COX-2.

Production of ethanol extract and propionic anhydride extract of Benix nokitake fruit body according to the present invention First, the fruit body of Benix nokitake fruit body is dried by a freeze-drying method and then freeze-dried at 42-45 ° C. 580 g of fruiting bodies are extracted with 95% ethanol to obtain the ethanol extract (ACE) of the present invention. This drying method is not limited, and any drying method known in the field of affiliation may be used.

The ethanol extract was then concentrated in a vacuum (rotary evaporator) to give 183.9 g of concentrated product. The obtained concentrated product is separated with propionic anhydride and water into a propionic anhydride layer and a water-soluble layer, and the water-soluble layer is labeled as an aqueous fraction (ACE-water), and the propionic anhydride is obtained. The layer is thus the propionic anhydride extract (ACE-EA) of the present invention.

Inhibition test of nitric oxide activity by ethanol extract (ACE), propionic anhydride extract (ACE-EA) and water-soluble fraction (ACE-water) of Example 2: Macrophage strain RAW 267.4
In this example, macrophage strain RAW 267.4 was used to extract the fruit body ethanol extract (ACE), propionic anhydride extract (ACE-EA) and water-soluble fraction (ACE) obtained in Example 2 -Water)) to inhibit the activity of nitric oxide.

First, RAW 267.4 cells were cultured by the cell culture method described in Example 1 and then experimented with Experiment Group 1 (ethanol extract, ACE) and Experiment Group 2 (propionic anhydride extract, ACE-EA). Divide into 3 groups (water-soluble fraction, ACE-water). According to the concentration (1, 10, 25, 50 g / ml) of each set of cells, the ethanol extract, the propionic anhydride extract and the water-soluble fraction are each put into a culture solution, and then lipopolysaccharide (LPS) Is added to the culture medium to induce RAW 267.4 cells to produce nitric oxide. After 24 hours, the nitrite content is measured by the grease reaction method, and the amount of nitric oxide produced Is measured indirectly, and the experimental design is shown in Table 1 below.

“+” Indicates addition, “−” indicates no addition.
The figure shows the statistical results of the amount of nitric oxide produced and the rate of inhibition of nitric oxide production measured by the grease reaction method. As can be seen from the figure, the Benix nokitake ethanol extract and the propionic anhydride extract obtained from Example 2 both have an activity suppressing effect on nitric oxide, particularly at a concentration of 50 g / ml. Becomes more prominent.

Nitric Oxide Activity Inhibition Test with Ethanol Extract (ACE) of Example 2: Mouse Living Body Experiment This example is a mouse living body, and nitric oxide activity inhibition with the ethanol extract (ACE) obtained in Example 2 Perform the test. As the mouse, an experimental mouse under the conditions described in Example 1 is used.

[Experimental design]
First, six mice are grouped, and there are six groups in total. According to the arrangement shown in Table 2 below, before injection of lipopolysaccharide (5 g / kg), each group of mice was injected intraperitoneally with an intraperitoneal injection method. Injected or not injected with Benix nokitake ethanol extract (ACE, 100, 300, 500 mg / kg), curcumin obtained in different Example 2 in which dimethyl sulfoxide ( DMSO) alone, and curcumin, known as an anti-inflammatory drug, is used in the positive control set in this example. Mice were injected with lipopolysaccharide and then anesthetized with ether, and after 12 hours, sacrificed by decapitation.

“+” Indicates addition, “−” indicates no addition.
Lipopolysaccharide was not administered to the control group, but dimethyl sulfoxide (DMSO) was administered instead.

[Experimental result]
After blood collection by the orbital blood collection method or the heart blood collection method, the blood is poured into a test tube containing ethylenediaminetetraacetic acid (EDTA), and the serum is collected by centrifugation at 500 g for 10 minutes at a temperature of 4 ° C. Serum nitric oxide concentration was measured, and the results are shown in Table 3 below.

According to the results in Table 3, the ethanol extract of Example 2 has a remarkable effect in suppressing the amount of nitric oxide produced in biological experiments, which is similar to the effect of curcumin.
Then, the liver of the sacrificed experimental mouse was removed, and the total protein was extracted by a well-known experimental method, and the protein concentration was determined by the Bradford method using a 595 nm absorbance value. Is used to measure the amounts of nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) produced by Western blot analysis (Western blot analysis, see Western blot analysis description in Example 1). As can be seen from the display results of FIG. 2A and FIG. 2B, the ethanol extract of Example 2 significantly reduced the production amounts of nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) in the living body experiment. However, it is considered that the ethanol extract of Example 2 suppressed the production of nitric oxide.

Separation of Antrocamphin A The propionic anhydride extract (ACE-EA) obtained in Example 2 was separated into 17 fractions by chromatographic method using a fluid phase concentration gradient. The solid phase used is silicon (60-80 mesh) is a fluid phase n-hexane / propionic anhydride mixture. The concentration gradient is such that the volume percentage of n-hexane and propionic anhydride is 95: 5, 90:10, 85:15, 80:20, 70:30, 60:40, 50:50, 40:60, 0: The n-hexane / propionic anhydride mixed solution in the order of 100 is used as the fluid phase by the chromatographic method.

About 17 obtained fractions, the activity suppression test to nitric oxide is performed with RAW267.4 cells (see the contents of cell culture in Example 1). First, RAW267.4 cells were divided into 17 groups, and the 17 fractions (50 g / ml) were added to the culture medium, respectively, and then lipopolysaccharide (LPS, 1 g / ml) was added to produce nitric oxide. After 24 hours, the content of nitrite is measured by the grease reaction method and the amount of nitric oxide produced is indirectly measured to calculate the inhibition rate of nitric oxide production. . As shown in FIG. 3, the experimental results show that the first, seventh, and tenth fractions have a suppression rate of generation of nitric oxide of 87%, 48%, and 64%, respectively.

Next, the first fraction is further separated and purified by high performance liquid chromatography (HPLC), and its components (solid phase: silicon tube column, fluid phase: volume ratio 85:15 n-hexane / propionic anhydride) Mixture, flow rate: 3 mm / min, UV wavelength: 254 nm) and the highest content component was analyzed spectrophotometrically to give a molecular weight of 247.14, which has the molecular formula C ₁₅ H ₁₉ O ₃ and Estimated. When the component is further analyzed by proton nuclear magnetic resonance spectroscopy, its molecular structure is as follows:

The compound is an antrocampin A.I. It was revealed that.

Example 6: Inhibition test of pro-inflammatory molecule production by Antrocamphin A: Macrophage strain RAW 267.4
In this example, a macrophage strain RAW 267.4 is used to conduct an activity suppression test of pro-inflammatory molecule production by Antrocamphin A obtained in Example 5.

The experiment on the amount of nitric oxide produced was carried out by first culturing RAW267.4 cells by the cell culture method described in Example 1, followed by different concentrations of antrocamphin A (1, 5, 10, 20 g / ml) or curcumin ( 10 g / ml) and left for 1 hour. After adding lipopolysaccharide (1 g / ml) and left for 24 hours, the content of nitrite is measured by the grease reaction method (nitrogen monoxide), and nitric oxide The production amount of is indirectly measured.

In the experiment of the amount of prostatic element E ₂ (PGE ₂ ) produced, first, RAW267.4 cells were cultured by the cell culture method described in Example 1, and then aspirin 500 m was added to the culture solution and left for 3 hours. Endogenous cyclooxygenase 1 (COX-1) activity is lost. Then, add different concentrations of antrocampin A (1, 5, 10, 20 g / ml) or curcumin (10 g / ml) and let stand for 1 hour, then add lipopolysaccharide (1 g / ml) and let stand for 16 hours Finally, in the enzyme immunosorbent assay (ELISA kit, Cayman Chemicals), the production of prostatic element E ₂ (PGE ₂ ) generated from endogenous arachidonic acid from the supernatant in cell culture Measure the amount.
Table 4 shows the groups tested in this example.

“+” Indicates addition, “−” indicates no addition.
Lipopolysaccharide was not administered to the control group, but dimethyl sulfoxide (DMSO) was administered instead.

In addition, the cytotoxicity of Antrocamphin A is measured by the well-known MTT method. The MTT method is a method often used in biology to measure cell viability or proliferation thereof. As a principle, MTT metabolites are metabolized by the action of succinate dehydrogenase in mitochondria in living cells. Is converted into a blue product, dimethyl sulfoxide (DMSO) is added to dissolve the blue metabolite deposited on the cells, and then the amount of blue metabolite is measured with a spectroscope. The number of cells can be estimated indirectly.
The experimental results are summarized in Table 5 below.

As can be seen from this result, antrocampin A can reduce the production of pro-inflammatory molecules, but the concentration at which a significant effect is exerted is 20 g / ml, and under this concentration, antrocamphin A has no occurrence of cytotoxicity. I can say that.

In addition, total ribonucleic acid (total RNA) and total protein extracted from each set of cells were analyzed using the reverse transcription-polymerase chain reaction and Western blot method described in Example 1 to obtain genetic material (mRNA) and protein. From these viewpoints, the amounts of nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) produced are evaluated.

4A and 4B show the amount of messenger ribonucleic acid (mRNA) produced by nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2), respectively. As can be seen from the results, the amount of messenger ribonucleic acid (mRNA) produced by nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) and antrocamfin A are related to the drug dose, and the high dose of antrocamffin A can effectively reduce the amount of messenger ribonucleic acid (mRNA) of nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2).

5A and 5B show the production amounts of nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2), respectively. As can be seen from the figures, nitric oxide synthase (iNOS) And cyclooxygenase 2 (COX-2) production amount and antrocamphin A are related to the drug dose, and high dose of antrocamphin A is the production amount of nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) Can be effectively reduced, but the effect is particularly noticeable at a concentration of 20 g / ml.

The results are summarized as follows. Antrocampin A reduces the production of nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) from the viewpoint of genetic material transcription or protein, and further induces inflammation. It has the effect of suppressing the production of sex molecules such as nitric oxide and prostate element E ₂ (PGE ₂ ).

Claims (19)

An extract of a fruit body of Benicus mushroom obtained by a procedure of providing a fruiting body of dried Benicum edulis mushroom and extracting the fruiting body of dried Benicococcus communis with ethanol at a constant temperature. 2. The extract of Benicus mushroom fruit body according to claim 1, wherein the drying method is freeze-drying. The extract of Benicaria mushroom fruit bodies according to claim 1, wherein the temperature is 42 to 45 ° C. The extract of a fruit body of Benicus mushroom according to claim 1, wherein the ethanol is 95% ethanol by volume. A fruit body of the dried venomus mushroom is provided, and the fruit body of the dried moss is extracted at a constant temperature to obtain an ethanol extract. The ethanol extract is concentrated and concentrated. An extract of Benicus japonica fruiting body characterized in that it is an extract of the propionic anhydride obtained by a procedure of partitioning the concentrated product with propionic anhydride and water. 6. The extract of Benicus mushroom fruit body according to claim 5, wherein the drying method is performed by freeze drying. 6. The extract of Benicus mushroom fruiting body according to claim 5, wherein the concentration is performed by vacuum concentration. The extract of Benicaria mushroom fruiting body according to claim 5, wherein the temperature is 42 to 45 ° C. 6. The extract of Benicaria mushroom fruiting body according to claim 5, wherein the ethanol is ethanol with a volume percentage of 95%. 6. The extract of Benicus mushroom fruit body according to claim 5, wherein the propionic anhydride is formed with propionic anhydride having a volume percentage of 50%. The use of the extract of Benicaria mushroom fruiting body according to any one of claims 1 to 10, which suppresses the production of pro-inflammatory molecules. The extract, the use of extracts of Benikusunokitakeko entity according to claim 11, characterized in that to suppress proinflammatory molecule that nitrogen monoxide (NO) or prostate element E _{2 (PGE 2).} 13. The extract according to claim 12, wherein nitric oxide synthase and cyclooxygenase 2 are reduced to suppress production of the nitric oxide (NO) and the prostatic element E ₂ (PGE ₂ ), respectively. Use of the extract of Benicus mushroom fruit body described. Use of antrocamphin A characterized by suppressing the production of nitric oxide (NO). The use of antrocamffin A according to claim 14, wherein the antrocampin A suppresses the production of the nitric oxide (NO) by decreasing nitric oxide synthase. Use of antrocamphin A characterized by suppressing the production of prostatic element E ₂ (PGE ₂ ). The use according to claim 16, wherein the antrocampin A reduces cyclooxygenase 2 and suppresses the production of the prostatic element E ₂ (PGE ₂ ). A method for inhibiting the production of pro-inflammatory molecules, comprising contacting a subject with any one of the extracts according to claim 17 or an antrocampin A. The inflammation-induced molecule A method according to claim 18, characterized in that the nitrogen monoxide (NO) or prostate element _E 2 (PGE _2).