CN112843177A

CN112843177A - Application of anoectochilus formosanus extract in preparation of medicine or health-care product for treating or relieving pain

Info

Publication number: CN112843177A
Application number: CN202110209282.6A
Authority: CN
Inventors: 郑承剑; 施艺; 吴岩斌; 吴锦忠; 王宏瑞; 周庆澳林; 张成中; 何旭辉
Original assignee: Second Military Medical University SMMU
Current assignee: Second Military Medical University SMMU
Priority date: 2021-02-24
Filing date: 2021-02-24
Publication date: 2021-05-28

Abstract

The invention discloses an application of an anoectochilus formosanus extract in preparation of a medicine or a health-care product for treating or relieving pain. The experimental result shows that the anoectochilus formosanus alcohol extract, the anoectochilus formosanus polysaccharide and the labyrin glycoside monomer compound have good in-vivo analgesic activity, have obvious inhibition effect on acute or chronic pain caused by chemical stimulation and thermal stimulation, can effectively reduce the times of writhing of mice induced by intraperitoneal injection of acetic acid, and prolong the pain response time of the mice; the pain threshold of the mouse in the thermal tail flick experiment is improved. The results indicate that the anoectochilus formosanus extract can be used for clinically treating various pains, especially inflammatory pains, or prepared into health-care products for relieving pains.

Description

Application of anoectochilus formosanus extract in preparation of medicine or health-care product for treating or relieving pain

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to an application of a high anoectochilus formosanus extract in preparation of a medicine or a health-care product for treating or relieving pain.

Background

Pain is the sensation produced when the body is subjected to various noxious stimuli, often accompanied by a number of diseases and injuries. The military is increasingly engaged in war military operations and executes diversified emergency tasks, and after a wounded person appears in the operation, analgesic drugs with strong analgesia and small side effect are selected, so that the life safety of the wounded person can be ensured, the pain of the wounded person can be reduced, and the strong stress response of an organism can be avoided. At present, the most clinical analgesic mode is drug analgesia, and the commonly used analgesic drugs are mainly nonsteroidal and opioid analgesics which have better curative effect clinically but have adverse reactions of different degrees. Therefore, the search and development of new, safe and effective analgesic drugs has been an important task in pain treatment.

"Cordyceps militaris is in north and anoectochilus formosanus is in south". Anoectochilus roxburghii is a special Chinese herbal medicine in Fujiatai, is derived from the Kalimerian odorifera Blume plant leaves of Kalimerian of Orchidaceae (Orchidaceae), is used as a medicine by whole herbs, and has the effects of clearing heat and cooling blood, dispelling wind and promoting diuresis, detoxifying and the like. The anoectochilus formosanus mainly contains lactone glycosides (such as anoectochilus formosanus glycoside), polysaccharides and flavonoid active ingredients, and is mainly used for treating hypertension, diabetes, hepatitis, tumors and other diseases (Zhanghongyan, Panxin. anoectochilus formosanus chemical ingredients and pharmacological activity research progress. Haixian pharmacy, 2009,21,82-84), and has a good curative effect, so the anoectochilus formosanus has the reputations of king and gold grass among people. As a medicine and food dual-purpose plant, anoectochilus formosanus has great demand and tends to exhaust wild resources, so that a plurality of confuses and counterfeit products appear in the market. Especially, some kindred species with the plant morphology similar to Anoectochilus formosanus, such as Zhejiang Anoectochilus formosanus, Xingren Anoectochilus formosanus, Nandan Anoectochilus formosanus, etc., are often mixed with Anoectochilus formosanus in folk, which seriously affects the clinical efficacy and safety of the Anoectochilus formosanus.

These confusable varieties were considered to contain the major component of Anoectochilus roxburghii glycoside (3- (R) -3- β -d-glucopyranosyloxybutanol, kinsenoside) according to the conventional high performance liquid detection method, but it was found by the optimized detection method that the major component contained in some of the confusable and counterfeit products was cantharidin (3- (S) -3- β -d-glucopyranosyloxybutanol, gooderoside A), i.e., Anoectochilus roxburghii glycoside isomer (Wu YB, Peng MC, Zhang C, Wu JG, Ye BZ, Yi J, Wu JZ, ZHehe CJ. quantitative determination of multi-class biological reagents for quality of animal of choice, Goodyearia strain and luminescence, 430, China flavor of Chinese family, China tea family, 439, 430). Anoectochilus formosanus glycoside and orychophragmus maculatus glycoside are epimers, and only lactone aglycone has different hydroxyl configuration and very similar polarity. The main chemical components are different, so the drug effects are different, and the rationality of the mixed drug of the folk confusable variety and anoectochilus formosanus is questioned inevitably. In 2018, a cheilotus plant (also used as Anoectochilus formosanus) which is found in the commodity survey and the production area tracking survey of the Yunnan Anoectochilus formosanus and has a plant shape very similar to that of the Anoectochilus formosanus is identified as Anoectochilus formosanus and is Anoectochilus elatus Lindl, although the plant shape is similar to that of the Anoectochilus formosanus, the flower is large and inverted; the front part of the labial lobe of the flower is nearly parallel to the base part, and the labial lobe is similar to a triangle and can be obviously different from anoectochilus formosanus. At present, the tissue culture of the high anoectochilus formosanus is successfully realized, and the resource problem is solved. The anoectochilus formosanus is found for the first time in China as a new domestic record species (Zhenglixiang, Tianhuazhen, Wujinzhong, xu bao ling, Zheng Chengjian, Huang ze Hao, Wu yanbin.) A new record species of Chinese orchidaceae plants, namely anoectochilus formosanus, Chinese modern Chinese medicines, 2018,20(1): 14-16). Earlier studies showed that the main chemical component of high anoectochilus formosanus is cantharidin, not anoectochilus formosanus glycoside. Furthermore, the academia find that although Anoectochilus roxburghii glycoside has various biological activities such as liver protection, blood sugar reduction and blood fat reduction, the isomers of the Anoectochilus roxburghii glycoside have no activity (Xiaoang M, Liu T, Tan W, Ren H, Li H, Liu J, Cao H, Cheng Q, Liu X, Zhu H, Tuo Y, Wang J, Zhang Y. effects of kinsenoside, a potential immunological therapy drug for autoimmunity, on pharmaceutical cells/CD8+ T cells research in micro. Hepatology.2016,64(6): 2135-. To date, no pharmacological activity report has been made on the extract of oryzanoside and anoectochilus formosanus. There are only reports in India that the use of the golden thread orchid is relevant for the treatment of insect and snake bites (Sherif N A, Kumar T S, Rao M V. in Vitro regeneration by bacteria culture of Anoectochilus linealey, an enderged terrestrial jewel organ. in Vitro Cellular & development Biology-Plant,2016,52(1): 72-80). However, reports on the analgesic application of the anoectochilus formosanus and the extract and the monomeric compound of the orychophragmus maculatus glycoside are not found so far.

Disclosure of Invention

The invention aims to provide application of a high anoectochilus formosanus extract in preparation of a medicine or health-care product for treating or relieving pain.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

the invention provides an application of an anoectochilus formosanus extract in preparation of a medicine or health-care product for treating or relieving pain, wherein the anoectochilus formosanus extract is at least one of an anoectochilus formosanus alcohol extract, anoectochilus formosanus polysaccharide or a labyrin monomeric compound.

The structure of the dracocephalin (goodroside A) monomer compound is shown as follows:

the preparation method of the anoectochilus formosanus alcohol extract comprises the following steps:

extracting the golden thread orchid with 40-95% ethanol water under reflux (the material-liquid ratio of ethanol to golden thread orchid is 30:1), and concentrating the extracting solution under reduced pressure to obtain an extract without alcohol smell to obtain the golden thread orchid alcohol extract.

The content of the oryzanolum glycosides in the anoectochilus formosanus alcohol extract is more than 50%.

The preparation method of the high anoectochilus formosanus polysaccharide comprises the following steps:

(1) extraction: drying the medicinal material residues of the high anoectochilus formosanus subjected to ethanol water extraction until no alcohol smell exists, heating and boiling the medicinal material residues with water for extraction for 2-3 times, wherein the dosage of a solvent is about 8-10 times of the dosage each time, the extraction time is 1-2 hours each time, and combining the extracting solutions;

(2) alcohol precipitation: concentrating the above extractive solution to original volume of 1/4, cooling, slowly adding 95% ethanol, adjusting ethanol concentration to 80%, standing in 4 deg.C refrigerator overnight, suction filtering, repeatedly washing precipitate with anhydrous ethanol and acetone to obtain filtrate in light yellow;

(3) and (3) purification: dissolving the precipitate in water at a ratio of 8mg/ml, centrifuging, collecting supernatant, wrapping in 3500KD ready-to-use dialysis bag, placing the dialysis bag in running water overnight, cleaning the dialysis bag with double distilled water for 2-4 times (each time for 1 hr), collecting water solution in the dialysis bag, concentrating under reduced pressure, and drying to obtain high-gold thread orchid polysaccharide.

The content of the high anoectochilus formosanus polysaccharide is greater than or equal to 90%.

The anoectochilus formosanus polysaccharide mainly comprises mannose, rhamnose, glucose, galactose, xylose and arabinose, and the molar ratio of the anoectochilus formosanus polysaccharide to the arabinose is 1:0.14:5.19:0.75:0.13:0.46 respectively.

The preparation method of the dracocephalin monomeric compound comprises the following steps:

(1) extraction: fully suspending the anoectochilus formosanus alcohol extract, namely the extract, in water (the material-liquid ratio of the extract to the water is 10:3), carrying out ultrasonic treatment for 1-60 min, and centrifuging to obtain a supernatant; repeatedly suspending the precipitate in water, performing ultrasonic treatment for 1-60 min, and filtering to obtain a supernatant; repeating for at least 3 times, and mixing the supernatants;

(2) and (3) drying: and (4) freeze-drying the combined supernatant to obtain the cantharidin monomeric compound.

The pain in the medicament or health product for treating or relieving pain is especially inflammatory pain.

Due to the adoption of the technical scheme, the invention has the following advantages and beneficial effects:

the invention adopts acetic acid induced mouse writhing, mouse heat tail flicking and mouse formalin licking foot model (Zheng CJ, Huang BK, Han T, Zhang QY, Zhang H, Rhman K, Qin LP. antibiotic activity of the lipid soluble fraction from vitamin green peptides, 2010,48, 651) 658), observes the analgesic effect of the high Anoectochilus formosanus extract by stomach filling administration. The experimental result shows that the anoectochilus formosanus alcohol extract, the anoectochilus formosanus polysaccharide and the labyrin glycoside monomer compound have good in-vivo analgesic activity, have obvious inhibition effect on acute or chronic pain caused by chemical stimulation and thermal stimulation, can effectively reduce the times of writhing of mice induced by intraperitoneal injection of acetic acid, and prolong the pain response time of the mice; the pain threshold of the mouse in the thermal tail flick experiment is improved; and the alcohol extract of the anoectochilus formosanus and the cantharidin monomer compound can obviously reduce the foot licking time of phase II caused by formalin injection on the sole of a mouse, and the effect cannot be antagonized by naloxone, which indicates that the compound does not have side effects such as morphine receptor agonist addiction and the like. The results indicate that the anoectochilus formosanus extract (comprising anoectochilus formosanus alcohol extract, anoectochilus formosanus polysaccharide and labyrin monomeric compound) can be used for clinically treating various pains, especially inflammatory pains, or prepared into health-care products for relieving pains.

The invention provides a new source for searching new medicaments and health care products for treating or relieving pain.

Drawings

FIG. 1 is a schematic diagram of the chemical structure of a monomeric compound of goodroside A.

FIG. 2 is an HPLC chromatogram of an alcohol extract of Gomphrena.

FIG. 3 is an HPLC chromatogram of monosaccharide composition of monosaccharide control and Gomphrena globosa polysaccharide (A. monosaccharide control, 1. mannose; 2. rhamnose; 3. glucuronic acid; 4. galacturonic acid; 5. glucose; 6. galactose; 7. xylose; 8. arabinose; B. Gomphrena globosa polysaccharide; 1. mannose; 2. rhamnose; 5. glucose; 6. galactose; 7. xylose; 8. arabinose).

FIG. 4 shows the effect of Gomphrena extract on acetic acid-induced writhing frequency in mice, wherein vs. blank control group, p <0.05, p <0.01(CON, blank group; COX, indomethacin; PS, polysaccharide; EE, alcohol extract; GA, cantharidin; L/M/H, low/medium/high).

FIG. 5 shows the effect of Gomphrena extract on writhing latency in acetic acid-induced mice, wherein vs. blank control group, p <0.05, p <0.01(CON, blank group; COX, indomethacin; PS, polysaccharide; EE, alcohol extract; GA, cantharidin; L/M/H, low/medium/high).

FIG. 6 shows the effect of golden orchid extract on hot tail flick pain threshold in mice, vs. blank control group, p <0.05, p <0.01(CON, blank group; COX, indomethacin; PS, polysaccharide; EE, alcohol extract; GA, cantharidin; L/M/H, low/medium/high).

FIG. 7 is a graph showing the effect of Gomphrena extract on formalin-induced mouse phase I licking time, wherein vs. blank control, p <0.05, p <0.01(CON, blank; MOR, morphine; ANT, naloxone; PS, polysaccharide; EE, alcohol extract; GA, labyrin; L/M/H, low/medium/high).

FIG. 8 is a graph showing the effect of Gomphrena extract on formalin-induced mouse phase II licking time, wherein vs. blank control, p <0.05, p <0.01(CON, blank; MOR, morphine; ANT, naloxone; PS, polysaccharide; EE, alcohol extract; GA, labyrin; L/M/H, low/medium/high).

Detailed Description

In order to more clearly illustrate the invention, the invention is further described below in connection with preferred embodiments. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.

Example 1

Preparation of extract of Anoectochilus formosanus

1) Preparing the golden thread orchid alcohol extract:

100g of medicinal material of the anoectochilus formosanus (obtained by autonomously culturing tissue culture seedlings in the laboratory of the applicant) is subjected to conventional reflux extraction by using 3L of 95% ethanol water solution (the material-liquid ratio of ethanol to the anoectochilus formosanus is 30:1), the extracting solution is subjected to reduced pressure concentration to obtain an extract without alcohol smell, 20g of the anoectochilus formosanus alcoholic extract is obtained, the content of the cantharidin (anoectochilus formosanus glycoside isomer) in the anoectochilus formosanus alcoholic extract is 59% by liquid phase analysis, as shown in figure 2, and figure 2 is an HPLC chromatogram of the anoectochilus formosanus alcoholic extract. As can be seen from the figure, the golden thread orchid alcohol extract has less chromatographic peaks and less impurities, and the main component is the macula orchid glycoside.

2) Preparing high anoectochilus formosanus polysaccharide:

The content of the polysaccharide in the anoectochilus formosanus is 90% as measured by a phenol-sulfuric acid method, the monomer composition mainly comprises mannose, rhamnose, glucose, galactose, xylose and arabinose, the molar ratio of the mannose to the rhamnose to the glucose to the galactose is 1:0.14:5.19:0.75:0.13:0.46, as shown in figure 3, figure 3 shows an HPLC chromatogram of the monosaccharide composition of a monosaccharide reference substance and the anoectochilus formosanus polysaccharide (A. monosaccharide reference substance, 1. mannose, 2. rhamnose, 3. glucuronic acid, 4. galacturonic acid, 5. glucose, 6. galactose, 7. xylose, 8. arabinose, B. anoectochilus formosanus polysaccharide, 1. mannose, 2. rhamnose, 5. glucose, 6. galactose, 7. xylose and 8. arabinose). As can be seen from the figure, the monosaccharide composition of the high anoectochilus formosanus polysaccharide is mainly glucose, mannose and galactose, and only contains a small amount of arabinose, rhamnose and xylose, and the molar ratio is 5.19:1:0.75:0.46:0.14:0.13 respectively.

3) Preparation of the labyrinthine monomeric compound:

(1) extraction: fully suspending the alcohol extract of the anoectochilus formosanus, namely the extract, in water (the material-liquid ratio of the extract to the water is 10:3), performing ultrasonic treatment for 30min, and centrifuging to obtain a supernatant; suspending the precipitate in water repeatedly, performing ultrasonic treatment for 30min, and filtering to obtain supernatant; repeating for 3 times, and mixing the supernatants;

(2) and (3) drying: freeze drying the combined supernatant at 0 deg.C overnight to obtain labyrinthine glycoside monomer compound, and detecting content by HPLC-ELSD>95 percent. Bonding of¹H-NMR、¹³C-NMR、DEPT、¹H-¹Spectroscopic data such as H COSY, HSQC, HMBC and NOESY (Zhang Y, Xia Y, Lai Y, Tang F, Luo Z, Xue Y, Yao G, Zhang Y, Zhang J. effective synthesis and goodyoroside a by a chemical-enzymetic approach. molecules,2014Oct 22; 19(10): 16900-8), the chemical structure of the monomeric compound for identifying the labyrinthine glycoside is shown in FIG. 1, and FIG. 1 is a schematic chemical structure diagram of the monomeric compound of the labyrinthine glycoside A.

Example 2

Analgesic pharmacodynamic experiment of high anoectochilus formosanus extract

2.1. Acetic acid induced writhing test in mice

2.1.1 Molding and grouping

80 healthy ICR mice are selected and randomly divided into 8 groups, each group comprises 10 mice, and the groups respectively comprise a blank group, a positive drug (indometacin) group, three groups with high, medium and low dosages of the anoectochilus formosanus polysaccharide and three groups with high, medium and low dosages of the anoectochilus formosanus alcohol extract.

2.1.2 dosage and administration

Each group of mice is infused with 0.1mL/10g of medicine every day, the positive medicine group is infused with 5mg/kg of indometacin solution, the high, middle and low dose groups of the experimental group are respectively infused with 25mg/kg, 50mg/kg and 100mg/kg of stomach, and the blank group is infused with physiological saline with the same volume as the stomach, and the total time is 7 days.

2.1.3 Observation index

After the last administration for 30min, each group was injected intraperitoneally with 0.6% acetic acid solution 0.2 mL/tube. The number of writhing reactions (belly concave, body and hind leg stretched, hip raised, wriggling) animals, the time of first writhing and the number of writhing of the mouse within 20min were observed.

2.2. Mouse thermal tail flick test

2.2.1 Molding and grouping

2.2.2 dosage and administration

Each group of mice is infused with 0.1mL/10g of medicine every day, the positive medicine group is infused with 5mg/kg of indometacin solution, the high, middle and low dose groups of the experimental group are respectively infused with 25mg/kg, 50mg/kg and 100mg/kg of stomach, and the blank group is infused with physiological saline with the same volume as the stomach.

2.2.3 Observation index

Vertically immersing the lower part of the tail of a mouse into a constant-temperature water bath kettle at 55 ℃, taking the latent period of tail retraction out of the water surface as a pain measurement index, measuring 2 times at an interval of 5min before administration, taking the average value as the basic pain threshold, measuring two sides after administration, and taking the average value as the pain threshold after administration. The pain threshold increase rate on the day of administration was measured separately.

2.3. Formalin-induced foot licking test for mice

2.3.1 Molding and grouping

140 healthy ICR mice are selected and randomly divided into 14 groups, each group comprises 10 mice, namely a blank group, a positive drug + antagonist group, three groups of high anoectochilus formosanus alcohol extract, high anoectochilus formosanus polysaccharide and high, medium and low doses of the cantharidin monomer compound, and a high dose of the cantharidin monomer compound + antagonist group.

2.3.2 dosage and administration

5mg/kg morphine is injected into the tail vein of the positive drug, 5mg/kg +0.4mg/kg naloxone is injected into the positive drug and antagonist group, 25mg/kg, 50mg/kg and 100mg/kg gastric lavage is respectively performed into the high, middle and low dose groups of the experimental group, equal volume of physiological saline is perfused into the blank group, 100mg/kg zebra floridin monomer compound is perfused into the high dose and naloxone group, 0.4mg/kg naloxone is injected into the tail vein, the administration volume is 1ml/100g per day, and the total 7 days are counted.

2.3.3 Observation index

30 mu L of 2.5% formalin solution (0.92% formaldehyde) is injected subcutaneously into the right hind paw after 30min of the last administration, and the right hind paw is immediately placed in a transparent container; and observing and recording the licking (biting) time of the mouse in 2 different stages of 0-5 min (I phase reaction) and 15-30 min (II phase reaction), and taking the time as an index of the animal pain reaction intensity.

2.4 results of the experiment

Two-sample t-test statistical analysis was performed on both sets of data using SPSS 22.0, and differences were considered statistically significant when P < 0.05.

2.4.1 Effect of Hypericum perforatum extract on acetic acid induced writhing frequency and latency of mice

As shown in FIG. 4, FIG. 4 shows the effect of the extract of Gomphrena on the number of writhing in acetic acid-induced mice, wherein vs. blank control group, p <0.05, p <0.01(CON, blank group; COX, indomethacin; PS, polysaccharide; EE, alcohol extract; GA, cantharidin; L/M/H, low/medium/high). As can be seen from the figure, after 7d of administration, the high anoectochilus formosanus alcohol extract, the high anoectochilus formosanus polysaccharide and the labyrinoside monomer compound administration groups can inhibit the mouse writhing frequency induced by intraperitoneal injection of acetic acid in a dose-dependent manner, and all have significant difference (p is less than 0.05) from the model group. The times of writhing of mice in the high-dose groups of the goldenlarch alcohol extract, the goldenlarch polysaccharide and the labyrin monomeric compound are respectively 5.4 +/-2.8, 4.4 +/-2.8 and 4.2 +/-2.9, which are equivalent to the times of writhing of indometacin group as a positive control drug (4.1 +/-1.9). The first writhing appearance time (latency) of mice in each administration group is also obviously prolonged, and is different from that of a model group (p <0.05) (as shown in figure 5, figure 5 shows the influence of the anoectochilus formosanus extract on the writhing latency of the mice caused by acetic acid, wherein vs. blank control group, p <0.05, p <0.01(CON, blank group, COX, indometacin, PS, polysaccharide, EE, alcohol extract, GA, labyrin, L/M/H, low/medium/high) are shown in the figure, wherein the latency time (minutes) of the labyrin monomer compound high dose group is equivalent to that of the positive drug group, and is respectively 6.0 +/-2.7 and 6.2 +/-2.3.

2.4.2 Effect of high Anoectochilus roxburghii extract on pain threshold of Hot Tail flicking mice

As shown in FIG. 6, FIG. 6 shows the effect of the extract of Gomphrena rostratum on pain threshold in hot tail flicking mice, wherein vs. blank control, p <0.05, p <0.01(CON, blank; COX, indomethacin; PS, polysaccharide; EE, alcohol extract; GA, cantharidin; L/M/H, low/medium/high). As can be seen from the figure, the increase rate of the pain threshold of the mice after administration is measured on the day of administration, and the pain threshold of the mice can be obviously increased by using the goldenlarch alcohol extract, the high-dosage group of the goldenlarch polysaccharide and the dosage group of the labyrin monomeric compound, and the pain threshold is obviously different from that of a blank control group (p is less than 0.05). The improvement rates of the high-dose groups of the anoectochilus formosanus alcohol extract, the anoectochilus formosanus polysaccharide and the labyrin monomer compound on the pain threshold value of a mouse are respectively 20.0%, 16.1% and 22.1%, and the improvement rates of the positive drug group are 30.9%.

2.4.3 Effect of high Anoectochilus roxburghii extract on formalin-induced mouse licking time

As shown in FIG. 7, FIG. 7 is a graph showing the effect of the extract of Gomphrena on the time of formalin-induced mouse phase I licking, wherein vs. blank control, p <0.05, p <0.01(CON, blank; MOR, morphine; ANT, naloxone; PS, polysaccharide; EE, alcohol extract; GA, labyrin; L/M/H, Low/Medium/high). As can be seen from the figure, in the I-phase reaction, the time (seconds) for the mice to lick the feet in the model group is 55.5 +/-8.9, and except for positive control morphine (24.4 +/-6.4), the high-dose group (45.4 +/-9.3) only containing the zebra compound monomer compound can obviously reduce the time for the mice to lick the feet, and has obvious difference (p <0.05) with the model group. In the II-phase reaction, as shown in FIG. 8, FIG. 8 is a graph showing the effect of the extract of Gomphrena on the formalin-induced paw licking time of mouse II-phase, where vs blank control group, p <0.05, p <0.01(CON, blank group; MOR, morphine; ANT, naloxone; PS, polysaccharide; EE, alcohol extract; GA, zebrinin; L/M/H, low/medium/high). As can be seen from the figure, the foot licking time (seconds) of the mice in the model group is 138.9 +/-23.7, and the low, medium and high dose groups of the high anoectochilus formosanus alcohol extract and the cantharidin monomeric compound can effectively reduce the foot licking time of the mice, are in a dose-dependent relationship and have a significant difference (p is less than 0.05) with the model group. Wherein, except morphine group (36.9 + -15.1), the high dose group of the zebra floridin monomer compound has the best effect (80.1 + -31.1). The single administration of naloxone has no obvious influence on the mouse licking time (140.2 +/-25.3), but the combined administration of naloxone and morphine (143.3 +/-29.1) can obviously antagonize the analgesic activity of morphine, but the combined administration of naloxone and a monodentate monomeric compound (95.3 +/-29.6) can not antagonize the analgesic activity of the monodentate, which indicates that the monodentate monomeric compound does not play an analgesic role through an opioid receptor, and suggests that the naloxone and the monodentate monomeric compound possibly have no side effects such as morphine receptor agonist addiction and the like, and the safety is better.

In view of the pharmacodynamic experiment results, the homoeoorchid extract (comprising the homoeoorchid alcohol extract, the homoeoorchid polysaccharide and the labyrin monomeric compound) has good in-vivo analgesic activity, has a remarkable inhibiting effect on acute or chronic pain caused by chemical stimulation and thermal stimulation, and prompts that the homoeoorchid extract (comprising the homoeoorchid alcohol extract, the homoeoorchid polysaccharide and the labyrin monomeric compound) can be used for clinically treating various pains, especially inflammatory pain, or prepared into health-care products for relieving pains.

Although the present invention has been described with reference to a preferred embodiment, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. The application of the anoectochilus formosanus extract in preparing the medicine or the health-care product for treating or relieving pain is characterized in that the anoectochilus formosanus extract is at least one of an anoectochilus formosanus alcohol extract, anoectochilus formosanus polysaccharide or a labyrin monomeric compound.

2. The application of the anoectochilus formosanus extract in preparing a medicine or health-care product for treating or relieving pain according to claim 1, wherein the eupatorium odoratum glycoside monomer compound has the following structure:

3. the use of the anoectochilus formosanus extract in the preparation of a medicament or health product for treating or relieving pain according to claim 1, wherein the preparation method of the anoectochilus formosanus extract comprises the following steps:

extracting the anoectochilus formosanus by refluxing with 40-95% ethanol water solution, and concentrating the extracting solution under reduced pressure to obtain an extract without alcohol smell to obtain the anoectochilus formosanus alcohol extract.

4. The use of the extract of anoectochilus formosanus as claimed in claim 3 for the preparation of a medicament or health product for treating or relieving pain, wherein the content of labyrin in the alcohol extract of anoectochilus formosanus is > 50%.

5. The use of the extract of anoectochilus formosanus as claimed in claim 3 for preparing a medicine or health product for treating or relieving pain, wherein the preparation method of the polysaccharide of anoectochilus formosanus comprises the following steps:

6. The use of the extract of anoectochilus formosanus as claimed in claim 5 for preparing a medicament or health product for treating or relieving pain, wherein the polysaccharide content of the anoectochilus formosanus is greater than or equal to 90%.

7. The use of the extract of Anoectochilus formosanus in the preparation of a medicament or health product for treating or relieving pain according to claim 5, wherein the Anoectochilus formosanus polysaccharides mainly comprise mannose, rhamnose, glucose, galactose, xylose and arabinose, and the molar ratio of the Anoectochilus formosanus to the mannose is 1:0.14:5.19:0.75:0.13: 0.46.

8. The application of the anoectochilus formosanus extract in preparing the medicines or health products for treating or relieving pain according to claim 3, wherein the preparation method of the cantharidin monomer compound comprises the following steps:

(1) extraction: fully suspending the alcohol extract of the anoectochilus formosanus, namely the extract, in water, carrying out ultrasonic treatment for 1-60 min, and centrifuging to obtain a supernatant; repeatedly suspending the precipitate in water, performing ultrasonic treatment for 1-60 min, and filtering to obtain a supernatant; repeating for at least 3 times, and mixing the supernatants;