Application of mikania micrantha extract in preparation of medicine for antagonizing TRPA1 and / or TRPV1 receptor

Abstract

The invention discloses an application of a mikania micrantha extract in preparation of a medicine for antagonizing TRPA1 and / or TRPV1 receptors. The research shows that the mikania micrantha extract has the effect of antagonizing a TRPA1 receptor or a TRPV1 receptor, also has the activity of antagonizing the TRPA1 receptor and the TRPV1 receptor, and can improve the symptom reaction of a model animal constructed by a specific TRPA1 receptor stimulant or a TRPV1 receptor stimulant, block the combination of the stimulant and the receptor, and inhibit the stimulant from playing a biological effect. Meanwhile, both the volatile oil extract and the aqueous extract of the mikania micrantha have better antagonistic effects which are equivalent to those of positive drugs, and have better antagonistic activity as a transient receptor potential (TRP) receptor antagonist, so that more methods and bases are provided for preparing more natural, safe and efficient drugs for antagonizing TRPA1 and / or TRPV1 receptors.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology. More specifically, it relates to the use of Mikania micrantha extract in the preparation of drugs antagonizing TRPA1 and / or TRPV1 receptors. Background Technology

[0002] The transient receptor potential (TRP) protein family is a class of non-selective gated cation channels located on cell membranes, distributed in the peripheral and central nervous systems, and widely present in animals. They primarily mediate the transmission of sensory signals, regulate cellular calcium homeostasis, and influence development. They play a role in various sensory responses, such as those triggered by temperature, pressure, light, and chemical stimuli, and serve as important targets for pain. TRP channels play a crucial role in normal physiological processes such as signal transduction; channel lesions or acquired dysfunction can disrupt these physiological processes, leading to widespread pathological changes. Therefore, they are closely related to various disease states, such as lung diseases, gastrointestinal diseases, rheumatic and immune diseases, and neoplastic diseases. The TRP channel family has many members, and based on differences in structure and function, they are usually divided into several subfamilies, including TRPA (ankylin), TRPC (typical), TRPV (capsaicin), and TRPM (Melastatin).

[0003] TRPA1, as a cold-sensitive ion channel, is widely distributed in both nerve and non-nerve cells, and is sensitive to Ca2+. 2+ TRPA1 exhibits good permeability and can be activated by endogenous and exogenous compounds, cold stimulation, mechanical stimulation, and various inflammatory mediators. Activation of TRPA1 is closely related to the generation and conduction of cold sensation, pain modulation, and the regulation of inflammatory substances. Furthermore, TRPA1 is associated with hypersensitivity and overexcitation in certain non-neuronal regions and plays an important role in the pathophysiology of respiratory diseases, neuropathic pain, digestive diseases, chronic pruritus, migraines, gastrointestinal motility disorders, anxiety, and cognitive impairment. It also participates in the inflammatory regulation of diseases such as inflammatory skin diseases, chronic obstructive pulmonary disease, bronchial asthma, and pancreatitis.

[0004] TRPV1 channels are ion channels located on cell membranes, lacking typical voltage receptors, and are primarily activated by stimulation from heat (>42°C), acidity (pH <6), capsaicin, and other plant-derived vanillins. They are widely expressed in tissues and cells of the cardiovascular, metabolic, digestive, respiratory, and urinary systems in mammals. Overactivation of TRPV1 receptors leads to a range of negative biological effects, including altered pain perception, increased inflammatory responses, and responses to certain disease states. In gastrointestinal malignancies, TRPV1 expression levels are associated with tumor progression and poor prognosis. In the nervous system, overexpression or activation of TRPV1 may affect neurogenesis. Overexpression of TRPV1 receptors is also associated with chronic pain.

[0005] Furthermore, TRPA1 and TRPV1 receptors are co-activated, and TRPV1 and TRPA1 channels are central to the inflammatory response and therapeutic targets for cough. Antagonists of TRPA1 and TRPV1 can reduce oxidative stress and block calcium channel blockers. 2+ The influx of substances inhibits inflammation and the high expression of antioxidant genes, and suppresses the activity of the mitochondrial respiratory chain (MRC) complex, thereby reducing the frequency of cough attacks. Clinically, there are few drugs that antagonize TRPA1 or TRPV1 receptors, and some antagonists have certain adverse reactions and side effects, such as overheating and burns. Currently, there are very few drugs that can simultaneously antagonize both TRPA1 and TRPV1 receptors.

[0006] Therefore, there is still a lack of effective drugs for antagonizing TRPA1 and / or TRPV1 receptors. It is necessary to develop more natural, safe and effective TRPA1 and / or TRPV1 antagonists, which is of great significance for the treatment of diseases caused by TRPA1 receptor or TRPV1 receptor, or by both TRPA1 and TRPV1 receptors. Summary of the Invention

[0007] The technical problem to be solved by the present invention is the lack of effective drugs for antagonizing TRPA1 receptor or TRPV1 receptor, as well as drugs that antagonize both TRPA1 and TRPV1 receptors. The present invention provides the application of Mikania micrantha extract in the preparation of drugs that antagonize TRPA1 and / or TRPV1 receptors.

[0008] The above-mentioned objective of this invention is achieved through the following technical solution:

[0009] This invention demonstrates that *Mikania micrantha* extract antagonizes TRPA1 or TRPV1 receptors, exhibiting simultaneous antagonistic activity against both. It improves symptom responses in animal models constructed with specific TRPA1 or TRPV1 receptor agonists, blocks the binding of agonists to receptors, and inhibits the biological effects of the agonists. Furthermore, studies show that both the volatile oil and water extracts of *Mikania micrantha* possess good antagonistic effects, comparable to those of positive control drugs. Since TRPA1 and TRPV1 receptors are crucial pathways for pain transmission, the *Mikania micrantha* extract, by simultaneously antagonizing the activity of both receptors, acts upstream in the pain signaling pathway, blocking pain signals and thus enabling its use in the preparation of analgesic drugs. The *Mikania micrantha* extract provided by this invention, as a transient receptor potential (TRP) receptor antagonist, exhibits good antagonistic activity and can be used to prepare therapeutic drugs for diseases induced by TRPA1 and / or TRPV1 receptor activation. This provides more methods and evidence for the preparation of more natural, safe, and highly effective drugs antagonizing TRPA1 and / or TRPV1 receptors.

[0010] Therefore, the present invention provides the use of Mikania extract as a transient receptor potential (TRP) receptor antagonist.

[0011] Furthermore, the present invention provides the use of Mikania extract as a transient receptor potential TRPA1 and / or TRPV1 receptor antagonist.

[0012] Preferably, the Mikania extract is obtained by distillation of Mikania.

[0013] This invention provides the application of Mikania micrantha extract in the preparation of transient receptor potential (TRP) receptor antagonist drugs.

[0014] Furthermore, the transient receptor potential (TRP) receptor is a TRPA1 receptor.

[0015] Furthermore, the transient receptor potential (TRP) receptor is a TRPV1 receptor.

[0016] Furthermore, the transient receptor potential (TRP) receptor is a TRPA1 receptor or a TRPV1 receptor.

[0017] Preferably, the Mikania extract is a Mikania volatile oil extract or a Mikania water extract.

[0018] More preferably, the preparation method of the Mikania micrantha volatile oil extract is as follows: take the whole Mikania micrantha herb, crush it, and extract it by steam distillation to obtain a brownish-yellow oily liquid as the Mikania micrantha volatile oil extract.

[0019] More preferably, the preparation method of the Mikania micrantha water extract is as follows: take the whole Mikania micrantha herb, crush it, perform steam distillation, filter the distilled water, concentrate it, and obtain a brownish-yellow thick extract as the Mikania micrantha water extract.

[0020] As the preferred embodiment, the present invention provides a more specific method for preparing Mikania micrantha volatile oil extract or Mikania micrantha aqueous extract:

[0021] Mikania micrantha volatile oil extract: Take the whole Mikania micrantha herb, crush it, add Mikania micrantha coarse powder, add about 8 times the amount of water, and extract by steam distillation for 6 hours to obtain a brownish-yellow oily liquid, which is the Mikania micrantha volatile oil extract.

[0022] Mikania micrantha aqueous extract: Take the whole herb of Mikania micrantha, crush it, add the coarse powder of Mikania micrantha, and then add about 8 times the amount of water. Extract by steam distillation for 6 hours. Filter the distilled water decoction, add about 8 times the amount of water to the residue, decoct for 1 hour, filter, combine the filtrates, concentrate, precipitate with 85% ethanol, let stand overnight, filter to remove the precipitate, recover the ethanol, concentrate and dry at below 60℃ to obtain a brownish-yellow extract, which is the Mikania micrantha aqueous extract.

[0023] This invention provides the use of Mikania micrantha extract as a transient receptor potential (TRP) receptor antagonist in the preparation of medicaments for treating diseases caused by activation of TRPA1 receptors and / or TRPV1 receptors.

[0024] Preferably, the disease is neuropathic pain, respiratory disease, inflammatory skin disease, chronic cough, neurodegenerative disease, digestive disease, cardiovascular disease, cancer, or pain disorder.

[0025] This invention also provides the use of Mikania extract in the preparation of analgesic drugs.

[0026] Furthermore, the drug also includes a pharmaceutically acceptable carrier or excipient.

[0027] Furthermore, the dosage form of the drug is a mixture, granules, tablets, capsules, drop pills, pills, ointments, tinctures, or injections.

[0028] The present invention has the following beneficial effects:

[0029] This invention demonstrates that *Mikania micrantha* extract antagonizes TRPA1 or TRPV1 receptors, and can simultaneously antagonize both. It improves symptom responses in animal models constructed with specific TRPA1 or TRPV1 receptor agonists, blocks the binding of agonists to receptors, and inhibits the biological effects of the agonists. Furthermore, the volatile oil and water extracts of *Mikania micrantha* exhibit good antagonistic effects, comparable to those of positive control drugs. As transient receptor potential (TRP) receptor antagonists, they demonstrate good antagonistic activity, providing more methods and evidence for the preparation of more natural, safe, and efficient drugs antagonizing TRPA1 and / or TRPV1 receptors. Attached Figure Description

[0030] Figure 1 The antagonistic effect of Mikania micrantha extract on TRPA1 receptor antagonism (x±s, n=10).

[0031] Figure 2 The antagonistic effect of Mikania micrantha extract on TRPA1 receptor antagonism (x±s, n=10). Detailed Implementation

[0032] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0033] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0034] In the following examples: HC030031 was purchased from Shanghai Maclean Biotechnology Co., Ltd., with a purity of 99%; isopropyl isothiocyanate was purchased from Shanghai Xianding Biotechnology Co., Ltd., with a purity of 98%; capsaicin was purchased from Shanghai Dibai Biotechnology Co., Ltd., with a purity of 98%; capsicumidine was purchased from Beijing Pusitang Biotechnology Co., Ltd., with a purity of 98%; Tween-80 and dimethyl sulfoxide were purchased from Shanghai Maclean Biotechnology Co., Ltd.

[0035] Example 1: Preparation of Mikania micrantha extract

[0036] 1. Vitex trifolia volatile oil extract

[0037] Take the whole herb of Mikania micrantha, crush it, add about 150g of Mikania micrantha coarse powder, then add about 8 times the amount of water, and extract by steam distillation for 6 hours to obtain a brownish-yellow oily liquid, which is Mikania micrantha volatile oil, for later use; the extraction rate of Mikania micrantha volatile oil is about 0.16%.

[0038] 2. Mikania micrantha water extract

[0039] Take the water decoction obtained after steam distillation of Mikania micrantha, filter it. Add about 8 times the amount of water to the medicinal residues, decoct for 1 h, filter, combine the filtrates, concentrate, perform alcohol precipitation with 85% ethanol, let it stand overnight, filter to remove the precipitate, recover ethanol, concentrate and dry at a temperature below 60 °C to obtain a brownish-yellow extract, which is the water extract of Mikania micrantha and is reserved; the extraction rate of the water extract of Mikania micrantha is about 5.58%.

[0040] Example 2 Blocking effect of Mikania micrantha extract on TRPA1 receptor

[0041] 1. Pharmacodynamic experiment method

[0042] Test animals: C57BL / 6 mice (purchased from Guangdong Medical Experimental Animal Center, animal certificate number: SCXK(Yue) 2022-0002), half male and half female, with a body weight of 18 - 24 g and an age of 6 - 8 weeks.

[0043] Test conditions: Temperature 22 °C ± 2 °C, relative humidity 65% ± 10%.

[0044] Test method: Take C57BL / 6 mice, half male and half female, weigh them, number them, randomly divide them into groups, with 10 mice in each group. Respectively set up a model group, a drug group, and a control group. After drug administration, place the mice in a transparent mouse cage for 30 min, observe the evoked responses of the model mice after subcutaneous injection of the TRPA1 receptor agonist (isopropyl isothiocyanate) on the nape of the neck, observe for 30 min, and calculate the inhibition rate of antagonizing the TRPA1 receptor according to the responses induced after the agonist binds to TRPA1.

[0045] Drug administration method: The positive drug (HC030031) and the test sample (different Mikania micrantha extracts) are administered by intraperitoneal injection; the TRPA1 receptor agonist (isopropyl isothiocyanate) is administered by subcutaneous injection, with a dose of 2.5 mg·kg -1 , and the above medicaments are usually administered at 50 μL.

[0046] Effect statistics: After injecting the TRPA1 receptor agonist, observe the evoked responses of mice in each group (after the TRPA1 receptor agonist activates the TRPA1 receptor, it will cause Ca 2+ influx, increasing intracellular Ca 2+The concentration of TRPA1 receptor agonists induces the release of neuropeptides, ultimately leading to neurogenic inflammation, respiratory inflammation, and hypersensitivity reactions. After binding to the TRPA1 receptor, TRPA1 receptor agonists induce acute and chronic itching, pain, inflammation, and allergic reactions. Animal models exhibit scratching behavior (positive control drugs can alleviate the induced reactions by antagonizing the TRPA1 receptor). The number of scratches was counted as one instance from when the mouse turned its head to touch the injection site until it turned back; licking lasting more than 3 seconds was counted as two instances; or lifting the hind paw to place it on the ground or licking it was counted as one instance. The inhibition rate of the TRPA1 receptor agonist was calculated using the following formula, and differential statistical analysis was performed.

[0047] Inhibition rate (%) = (Number of scratches in the blank control group - Number of scratches in the test group) / Number of scratches in the blank control group × 100%

[0048] 2. TRPA1 receptor model group

[0049] C57BL / 6 mice were used, and the hair on the back of their necks was shaved one day before administration. The next day, the shaved mice were injected intraperitoneally with 50 μL of physiological saline. After placing the mice in transparent cages for 30 minutes, isopropyl isothiocyanate was subcutaneously injected into the back of the neck. The isopropyl isothiocyanate solution was prepared with water to a concentration of 1 mg / mL (final concentration of DMSO, Tween-80 not exceeding 1%). The dosage for mice was 2.5 mg / kg. -1 Drug administration was performed to construct a TRPA1 receptor model group.

[0050] 3. Antagonist drug group

[0051] (1) Positive drug group: The test sample was prepared with HC030031 solution, with 0.5% PEG400 to prepare an 8 mg / mL solution, at 20 mg·kg -1 The drug is administered to the positive control group to exclude false negative results and to demonstrate the reliability and repeatability of the test method.

[0052] (2) High-concentration volatile oil group: The test sample was prepared using a high-concentration solution of Mikania micrantha volatile oil. 128 mg of volatile oil was dissolved in dimethyl sulfoxide (DMSO) and diluted to 10 mL to obtain a volatile oil stock solution of 12.8 mg / mL. An appropriate amount of the stock solution was then diluted with water to a solution of 0.128 mg / mL. Mice were administered the solution at a concentration of 0.32 mg / kg. -1 Administer the medication.

[0053] (3) Medium concentration group of volatile oil: The test samples were prepared using a medium concentration solution of Mikania micrantha volatile oil. An appropriate amount of the above volatile oil stock solution was taken and diluted with water to a concentration of 0.064 mg / mL. Mice were administered the solution at a concentration of 0.16 mg / kg. -1 Administer the medication.

[0054] (4) Low-concentration volatile oil group: The test samples were prepared using a low-concentration solution of Mikania micrantha volatile oil. An appropriate amount of the above volatile oil stock solution was diluted with water to a concentration of 0.032 mg / mL. Mice were administered the solution at a concentration of 0.08 mg / kg. -1 Administer the medication.

[0055] (5) High-concentration aqueous extract group: The test samples were prepared using a high-concentration solution of Mikania micrantha aqueous extract. An appropriate amount of Mikania micrantha aqueous extract was diluted with water to a concentration of 4.46 mg / mL. Mice were administered the solution at a concentration of 11.15 mg / kg. -1 Administer the medication.

[0056] (6) Medium concentration group of water extract: The test samples were prepared using a medium concentration solution of Mikania micrantha water extract. An appropriate amount of Mikania micrantha water extract was diluted with water to a concentration of 2.23 mg / mL. Mice were administered the solution at a concentration of 5.58 mg / kg. -1 Administer the medication.

[0057] (7) Low concentration group of water extract: The test samples were prepared using a low concentration solution of Mikania micrantha water extract. An appropriate amount of Mikania micrantha water extract was diluted with water to a concentration of 1.115 mg / mL. Mice were administered the solution at a concentration of 2.79 mg / kg. -1 Administer the medication.

[0058] 4. Control group

[0059] C57BL / 6 mice were used, and the hair on the back of their necks was shaved the day before drug administration. The next day, the shaved mice were taken, and 50 μL of physiological saline was injected intraperitoneally. After the mice were placed in transparent cages for 30 minutes, 50 μL of physiological saline was injected subcutaneously into the back of their necks as a negative control group.

[0060] 5. Antagonistic effect of Mikania micrantha extract on TRPA1 receptor

[0061] By constructing a TRPA1 receptor model using specific methods in this field (using a specific TRPA1 receptor agonist) and then treating the model mice with different Mikania extracts, the phenotypic response induced by the binding of the agonist to TRPA1 can be alleviated. The statistical results of the induced responses in each group of model mice are shown in Table 1 below.

[0062] Table 1. Antagonistic effect of Mikania micrantha extract on TRPA1 receptor ( n=10)

[0063]

[0064]

[0065] Based on the experimental results, a statistical analysis of the differences was performed, and the results are as follows: Figure 1As shown, the P value between the display model group and the positive drug group was <0.01, indicating a highly significant difference, suggesting successful model establishment; the P values between the high-concentration volatile oil group of Mikania micrantha, the medium-concentration volatile oil group and the model group were <0.01, and the P value between the high-concentration aqueous extract group of Mikania micrantha and the model group was <0.01, showing highly significant differences, which could well inhibit the induced response of model mice and block the action of the TRPA1 channel; the P value between the low-concentration aqueous extract group and the model group was <0.05, indicating a significant difference. Although the P values of the low-concentration volatile oil group and the medium-concentration aqueous extract group were >0.05 compared with the model group, the scratching times of the mice were still reduced compared with the model, still showing the effect of blocking TRPA1, indicating that the volatile oil and aqueous extract of Mikania micrantha have an antagonistic effect on the TRPA1 receptor.

[0066] Example 3 Blocking Effect of Mikania micrantha Extract on TRPV1 Receptor

[0067] 1. Pharmacodynamic Experiment Method

[0068] Experimental animals: C57BL / 6 mice (purchased from Guangdong Medical Experimental Animal Center, animal certificate number: SCXK(Yue)2022-0002), half male and half female, weighing 18-24 g, 6-8 weeks old.

[0069] Experimental conditions: temperature 22°C ± 2°C, relative humidity 65% ± 10%.

[0070] Experimental method: Take C57BL / 6 mice, half male and half female, weigh them, number them, and randomly divide them into groups of 10 each. Respectively set up a model group, a drug group, and a control group. After drug administration, place the mice in a transparent mouse cage for 30 min, and observe the induced response of the model mice after subcutaneous injection of the TRPV1 receptor agonist (capsaicin) on the back of the neck. The observation time is 30 min, and the inhibition rate of antagonizing the TRPV1 receptor is statistically analyzed according to the response caused by the binding of the agonist to TRPV1.

[0071] Drug administration method: The positive drug (capsazepine) and the test sample (different extracts of Mikania micrantha) were administered by intraperitoneal injection; the TRPV1 receptor agonist (capsaicin) was administered by subcutaneous injection, with a dose of 7.5 μg·kg -1 , and the above-mentioned drugs were usually administered at 50 μL.

[0072] Effect statistics: After injection of the TRPV1 receptor agonist, observe the induced response of each group of mice (activation of the TRPV1 receptor by the TRPV1 receptor agonist will cause Ca 2+ influx and increase intracellular Ca 2+Concentration ultimately leads to respiratory diseases, cognitive impairment, inflammatory skin diseases, digestive diseases, and cardiovascular diseases. After binding to the TRPV1 receptor, TRPV1 receptor agonists regulate pain transmission at sensory synapses, triggering an allergic reaction. Animal models exhibit scratching behavior (positive control drugs can alleviate the induced reaction by antagonizing TRPV1 receptors). The number of scratches was counted as one instance from when the mouse turned its head to near the injection site until it turned back; licking lasting more than 3 seconds was counted as two instances; or lifting the hind paw to place it on the ground or licking it was counted as one instance. The TRPV1 receptor agonist inhibition rate was calculated using the following formula, and differential statistical analysis was performed.

[0073] Inhibition rate (%) = (Number of scratches in the blank control group - Number of scratches in the test group) / Number of scratches in the blank control group × 100%

[0074] 2. TRPV1 receptor model group

[0075] C57BL / 6 mice were used, and the hair on the back of their necks was shaved one day before administration. The next day, the shaved mice were taken, and after an intraperitoneal injection of 50 μL of physiological saline, they were placed in transparent cages for 30 minutes. Then, capsaicin solution was subcutaneously injected into the back of the neck. The capsaicin solution was prepared with water to a concentration of 3 μg / mL (final concentration of DMSO not exceeding 1%). The dosage for mice was 7.5 μg / kg. -1 Drug administration was performed to construct a TRPV1 receptor model group.

[0076] 3. Antagonist drug group

[0077] (1) Positive drug group: The test sample was prepared with capsaicin solution, with 0.5% PEG400 to a concentration of 4 mg / mL, at a concentration of 10 mg / kg. -1 The drug is administered to the positive control group to exclude false negative results and to demonstrate the reliability and repeatability of the test method.

[0078] (2) High-concentration volatile oil group: The test sample was prepared using a high-concentration solution of Mikania micrantha volatile oil. 128 mg of volatile oil was dissolved in dimethyl sulfoxide (DMSO) and diluted to 10 mL to obtain a volatile oil stock solution of 12.8 mg / mL. An appropriate amount of the stock solution was then diluted with water to a solution of 0.128 mg / mL. Mice were administered the solution at a concentration of 0.32 mg / kg. -1 Administer the medication.

[0079] (3) Medium concentration group of volatile oil: The test samples were prepared using a medium concentration solution of Mikania micrantha volatile oil. An appropriate amount of the above volatile oil stock solution was taken and diluted with water to a concentration of 0.064 mg / mL. Mice were administered the solution at a concentration of 0.16 mg / kg. -1 Administer the medication.

[0080] (4) Low-concentration volatile oil group: The test samples were prepared using a low-concentration solution of Mikania micrantha volatile oil. An appropriate amount of the above volatile oil stock solution was diluted with water to a concentration of 0.032 mg / mL. Mice were administered the solution at a concentration of 0.08 mg / kg. -1 Administer the medication.

[0081] (5) High-concentration aqueous extract group: The test samples were prepared using a high-concentration solution of Mikania micrantha aqueous extract. An appropriate amount of Mikania micrantha aqueous extract was diluted with water to a concentration of 4.46 mg / mL. Mice were administered the solution at a concentration of 11.15 mg / kg. -1 Administer the medication.

[0082] (6) Medium concentration group of water extract: The test samples were prepared using a medium concentration solution of Mikania micrantha water extract. An appropriate amount of Mikania micrantha water extract was diluted with water to a concentration of 2.23 mg / mL. Mice were administered the solution at a concentration of 5.58 mg / kg. -1 Administer the medication.

[0083] (7) Low concentration group of water extract: The test samples were prepared using a low concentration solution of Mikania micrantha water extract. An appropriate amount of Mikania micrantha water extract was diluted with water to a concentration of 1.115 mg / mL. Mice were administered the solution at a concentration of 2.79 mg / kg. -1 Administer the medication.

[0084] 4. Control group

[0085] C57BL / 6 mice were used, and the hair on the back of their necks was shaved the day before drug administration. The next day, the shaved mice were taken, and 50 μL of physiological saline was injected intraperitoneally. After the mice were placed in transparent cages for 30 minutes, 50 μL of physiological saline was injected subcutaneously into the back of their necks as a negative control group.

[0086] 5. Antagonistic effect of Mikania micrantha extract on TRPV1 receptor

[0087] Studies using TRPV1 receptor models constructed using specific methods in this field (constructed from specific TRPV1 receptor agonists) have shown that treatment with Mikania micrantha extract in model mice can alleviate allergic reactions induced by the binding of the agonist to TRPV1. The statistical results of the induced responses in each group of model mice are shown in Table 2 below.

[0088] Table 2. Antagonistic effect of Mikania micrantha extract on TRPV1 receptor ( n=10)

[0089]

[0090] Based on the experimental results, statistical analysis of the differences was performed, such as... Figure 2As shown, the model group showed a significant difference (P<0.05) compared to the positive drug group, indicating successful modeling. The high-concentration groups of Mikania micrantha volatile oil and water extract showed highly significant differences (P<0.01) compared to the model group, indicating that the high-concentration Mikania micrantha extract better inhibited the induced response in the model mice and blocked the TRPV1 channel. The medium-concentration groups of volatile oil and water extract showed significant differences (P<0.05) compared to the model group. Combined with dosage analysis, the activity of the volatile oil was stronger than that of the positive drug. Although the low-concentration groups of volatile oil and water extract showed no significant difference (P>0.05), the number of scratches by the mice was still reduced compared to the model, indicating that TRPV1 activity was blocked, suggesting that Mikania micrantha volatile oil and water extract have antagonistic effects on the TRPV1 receptor.

[0091] In the above experimental groups, the dosages of the positive control drug, the high concentration of volatile oil, and the high concentration of water extract were different, but the differences among the three groups were not statistically significant (P>0.05). This indicates that both the volatile oil extract and the water extract of Mikania micrantha have good antagonistic effects, comparable to those of the positive control drug.

[0092] In summary, the results show that Mikania micrantha extract can block transient receptor potential (TRP) receptors such as TRPA1 and TRPV1, simultaneously antagonizing both receptors. It inhibits the induced response in mouse models constructed with specific TRP receptor agonists, reduces scratching frequency, and blocks the TRPA1 and TRPV1 channels. Furthermore, both the volatile oil and water extracts of Mikania micrantha exhibit good antagonistic effects. The antagonistic effect of the water extract is comparable to that of the positive control drug, and the antagonistic activity of the volatile oil is stronger than that of the positive control drug when considering dosage. Therefore, Mikania micrantha extract, as a transient receptor potential (TRP) receptor antagonist, possesses good antagonistic activity and can be used to prepare therapeutic drugs for diseases caused by activation of TRPA1 and / or TRPV1 receptors. This invention provides methods and a basis for preparing more natural, safe, and efficient drugs that antagonize TRPA1 and / or TRPV1 receptors.

[0093] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. The application of Mikania micrantha extract as a transient receptor potential TRPA1 and / or TRPV1 receptor antagonist, characterized in that, The Mikania extract is obtained by distillation of Mikania.

2. The application of Mikania micrantha extract in the preparation of transient receptor potential (TRP) receptor antagonist drugs, characterized in that, The Mikania extract is obtained by distillation of Mikania.

3. The application according to claim 2, characterized in that, The transient receptor potential TRP receptor is the TRPA1 receptor.

4. The application according to claim 2, characterized in that, The transient receptor potential TRP receptor is the TRPV1 receptor.

5. The application according to claim 2, characterized in that, The transient receptor potential (TRP) receptors are TRPA1 receptors and TRPV1 receptors.

6. The application according to any one of claims 1 to 5, characterized in that, The Mikania extract is either a Mikania volatile oil extract or a Mikania water extract.

7. The application according to claim 6, characterized in that, The preparation method of the Mikania micrantha volatile oil extract is as follows: take the whole Mikania micrantha herb, crush it, and extract it by steam distillation to obtain a brownish-yellow oily liquid, which is the Mikania micrantha volatile oil extract.

8. The application according to claim 6, characterized in that, The preparation method of the Mikania micrantha water extract is as follows: take the whole Mikania micrantha herb, crush it, perform steam distillation, filter the distilled water, concentrate it, and obtain a brownish-yellow thick extract as the Mikania micrantha water extract.

9. The use of Mikania extract as a transient receptor potential (TRP) receptor antagonist in the preparation of therapeutic drugs for diseases caused by activation of TRPA1 receptor and / or TRPV1 receptor.

10. The application according to claim 9, characterized in that, The diseases mentioned include neuropathic pain, respiratory diseases, inflammatory skin diseases, chronic cough, neurodegenerative diseases, digestive diseases, cardiovascular diseases, cancer, or pain disorders.

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