Extract of Chinese herbal medicine spanishneedles herb as well as extraction method and application of extract

Abstract

The invention belongs to the technical field of Chinese herbal medicine extracts, and discloses a Chinese herbal medicine spanishneedles herb extract and an extraction method and application thereof.The extraction method comprises the steps that S1, dried whole spanishneedles herb is cut into sections, soaked with purified water and then decocted and extracted, an extracting solution is filtered out, purified water is added for decoction and extraction, an extracting solution is filtered out, the extracting solutions are combined and concentrated, and concentrated liquid is obtained; a concentrated extracting solution is obtained; and S2, taking the concentrated extracting solution, adding ethanol, standing, precipitating, taking supernate, and concentrating to obtain a concentrated solution containing the extract of the Chinese herbal medicine spanishneedles herb. The extract is prepared into oral liquid, granules, capsules, tablets, powder injection or injection dosage form medicines. The extract is applied to preparation of drugs for treating hypertension, drugs for treating hypertension complications and foods for reducing blood pressure. Scientific basis and technical support are provided for application of spanishneedles herb in hypertension treatment, and a new thought is provided for preparation of safe and effective antihypertensive drugs, health care products or functional food.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine extract technology, specifically to an extract of the traditional Chinese medicine Bidens pilosa, its extraction method, and its application. Background Technology

[0002] Hypertension is a common chronic cardiovascular disease that accompanies daily life for a long time. It is characterized by persistently elevated systemic arterial blood pressure. Long-term hypertension can damage multiple organs, including the heart, brain, kidneys, and eyes, seriously threatening human health. Currently, commonly used antihypertensive drugs in clinical practice include calcium channel blockers, angiotensin-converting enzyme inhibitors, angiotensin II receptor antagonists, diuretics, and beta-blockers. While these drugs can effectively control blood pressure, long-term use may cause adverse reactions such as dizziness, fatigue, electrolyte imbalance, and liver and kidney damage. Therefore, exploring antihypertensive components from natural medicines that have stable effects, multi-target regulation, and relatively few side effects has become one of the important directions in current drug development.

[0003] Bidens pilosa L., a medicinal plant belonging to the genus Bidens in the family Asteraceae, is widely distributed throughout my country. In traditional medicine, Bidens pilosa is used to treat sore throat, traumatic injuries, and damp-heat jaundice, possessing properties that clear heat and detoxify, dispel wind and dampness, and promote blood circulation and reduce swelling. Modern research further reveals that Bidens pilosa is rich in flavonoids, polyphenols, and terpenes, exhibiting not only antioxidant, anti-inflammatory, and antibacterial pharmacological effects, but also potential value in regulating blood pressure and improving vascular function. Summary of the Invention

[0004] This invention aims to provide an extract of the traditional Chinese medicine Bidens pilosa, its extraction method, and its application, providing a scientific basis and technical support for the application of Bidens pilosa in the treatment of hypertension, and offering new ideas for the preparation of safe and effective antihypertensive drugs, health products, or functional foods.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] A method for extracting an extract from the traditional Chinese medicine Bidens pilosa includes the following steps:

[0007] S1. Take dried Bidens pilosa whole herb, cut it into sections, and soak it in 8-10 times the weight of the herb in purified water for 1-1.5 hours. Then decoct and extract for 1-2 hours, filter out the extract, add 8-10 times the weight of purified water and decoct and extract for 1-2 hours, filter out the extract, combine the two extracts and concentrate to obtain concentrated extract.

[0008] S2. Take the concentrated extract obtained in step S1, add ethanol to make the ethanol concentration reach 60-80%, let it stand for 24 hours, precipitate, take the supernatant, concentrate, and obtain an alcohol precipitation concentrate containing the extract of the Chinese herbal medicine Bidens pilosa.

[0009] A method for extracting an extract from the traditional Chinese medicine Bidens pilosa includes the following steps: taking dried Bidens pilosa whole herb and cutting it into sections, first extracting it by refluxing with 60% to 80% ethanol at 8 to 10 times the weight of the herb, and filtering out the extract; then extracting it by refluxing with 60% to 80% ethanol at 8 to 10 times the weight of the herb, filtering out the extract, combining the two filtrates, evaporating the ethanol, and concentrating to obtain a concentrated extract containing the traditional Chinese medicine Bidens pilosa.

[0010] The extract was obtained using the above-mentioned extraction method for the traditional Chinese medicine Bidens pilosa.

[0011] Further, starch or magnesium stearate is added to the concentrate and mixed. After drying, tablets are made by compression or granulation, cut into doses, and packaged to obtain tablets.

[0012] Further, excipients are added to the concentrate and mixed to obtain a soft material, which is then sieved through a gyratory granulator to obtain wet granules. The wet granules are then placed in a fluidized bed dryer or oven to dry. The dried granules are sieved using a granulator and then divided into doses and encapsulated to form capsules.

[0013] Furthermore, the excipient is starch or magnesium stearate.

[0014] Furthermore, the obtained concentrate is filtered through micropores, freeze-dried, and sterilized to produce a powder injection.

[0015] Further, distilled water and activated carbon were added to the concentrate, heated at 80°C for 30 minutes, coarsely filtered, the pH value was adjusted, then filtered through a microporous membrane, filled, and sterilized by high-temperature autoclaving for 30 minutes to obtain the injection solution.

[0016] The application of the extract obtained using the above-mentioned extraction method of the traditional Chinese medicine Bidens pilosa in the preparation of drugs for treating hypertension or drugs for treating complications of hypertension.

[0017] The application of the extract obtained using the above-mentioned extraction method of the traditional Chinese medicine Bidens pilosa in the preparation of blood pressure-lowering food products.

[0018] The beneficial effects of the technical solution are:

[0019] 1. The extraction method for Bidens pilosa provided by this invention comprises two methods: First, the water extraction and alcohol precipitation method (S1-S2) fully dissolves the hydrophilic active substances in Bidens pilosa by boiling in water, and then removes macromolecular impurities (such as polysaccharides and proteins) by ethanol precipitation, thereby enriching effective components such as flavonoids and polyphenols. This method is low-cost, highly compatible with traditional medication experience, and easy to promote and use. The ethanol reflux extraction method directly uses medium-concentration ethanol as a solvent, which can more efficiently extract both hydrophilic and moderately polar active components simultaneously. This method has a simple process and high extraction efficiency. Both methods, by optimizing key parameters such as solvent ratio (60-80% ethanol), extraction times, and time, can systematically extract flavonoids, polyphenols, and terpenes with antihypertensive potential from Bidens pilosa, and effectively remove ineffective impurities, improving the purity and consistency of the extract's pharmacodynamic material basis. This provides a feasible, stable, and scalable process option for the industrial extraction of Bidens pilosa extracts.

[0020] 2. The extract obtained by the extraction method of the herbal medicine Bidens pilosa provided in this invention is derived from nature. Compared with existing chemically synthesized antihypertensive drugs (calcium channel blockers, angiotensin-converting enzyme inhibitors, angiotensin II receptor antagonists, diuretics, β-blockers, etc.), the extract has the characteristics of multi-target and systemic regulation (taking into account anti-oxidation, anti-inflammation, and improvement of vascular function). While achieving the antihypertensive effect, it can effectively reduce or avoid typical side effects such as dizziness, electrolyte imbalance, and liver and kidney function effects caused by long-term use of chemical drugs, providing a safer and gentler supplementary or alternative treatment option for patients with hypertension.

[0021] 3. The extraction method of the herbal medicine Bidens pilosa provided by this invention yields extracts that can be formulated into tablets, capsules, powder injections, and injection solutions, thus expanding the product's applicability: tablets and capsules are convenient for oral administration and suitable for long-term management of hypertension; powder injections and injection solutions have a faster onset of action, providing new treatment options for hypertensive emergencies or clinical situations requiring rapid blood pressure control.

[0022] 4. The extraction method of the herbal medicine Bidens pilosa provided by this invention can be used to prepare drugs for treating hypertension and its complications (hypertension combined with kidney disease, hypertension caused by kidney disease, and renal hypertension caused by tumor drug treatment), as well as foods with blood pressure lowering function (such as health foods and functional foods). This provides a complete technical chain and legal basis for transforming Bidens pilosa, a traditional herbal resource, into products (pharmaceuticals, health products, or functional foods) with modern quality standards and clear uses. Attached Figure Description

[0023] Figure 1Venn diagram of Bidens pilosa extract and hypertension target in Example 3 of this invention;

[0024] Figure 2 This is a PPI network diagram of the Bidens pilosa extract used to treat hypertension, as shown in Example 3 of the present invention.

[0025] Figure 3 This is a diagram showing the relationship between the target sites of the Bidens pilosa extract in Example 3 of the present invention.

[0026] Figure 4 This is a GO functional enrichment map of the target site of the Bidens pilosa extract in Example 3 of the present invention.

[0027] Figure 5 This is a KEGG pathway enrichment map of the target site of the Bidens pilosa extract in Example 3 of the present invention.

[0028] Figure 6 This is a statistical graph showing the systolic blood pressure of hypertensive rats induced by different doses of Bidens pilosa in Example 4 of this invention.

[0029] In the figure, Control represents the normal control group, Model represents the anlotinib model group, Positive represents the positive control group, High-dose represents the high-dose Bidens pilosa group, Medium-dose represents the medium-dose Bidens pilosa group, and Low-dose represents the low-dose Bidens pilosa group.

[0030] Figure 7 This is a statistical graph showing the mean arterial pressure of hypertensive rats induced by different doses of Bidens pilosa in Example 4 of this invention.

[0031] In the figure, Control represents the normal control group, Model represents the anlotinib model group, Positive represents the positive control group, High-dose represents the high-dose Bidens pilosa group, Medium-dose represents the medium-dose Bidens pilosa group, and Low-dose represents the low-dose Bidens pilosa group. Detailed Implementation

[0032] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments:

[0033] Example 1: A method for extracting an extract from the traditional Chinese medicine Bidens pilosa, comprising the following steps:

[0034] S1. Take 1000 portions of dried Bidens pilosa whole herb, cut it into 1-2cm small sections, soak it in 8-10 times the weight of the herb in purified water for 1-1.5 hours, then decoct under low pressure for 1-2 hours, filter out the extract, add 8-10 times the weight of purified water and decoct under low pressure for 1-2 hours, filter out the extract, combine the two extracts and concentrate to obtain concentrated extract;

[0035] S2. Take the concentrated extract obtained in step S1, add ethanol to make the ethanol concentration reach 60-80%, let stand for 24 hours, precipitate, take the supernatant, concentrate, and obtain an alcohol precipitation concentrate containing the extract of the Chinese herbal medicine Bidens pilosa.

[0036] Example 2: A method for extracting an extract from the traditional Chinese medicine Bidens pilosa, comprising the following steps:

[0037] Take 1000 portions of dried Bidens pilosa whole herb, cut into 1-2 cm segments, and extract by reflux with 8-10 times the amount of the herb in 60%-80% ethanol. Filter the extract, then extract by reflux with 8-10 times the amount of the herb in 60%-80% ethanol. Filter the extract, combine the two filtrates, evaporate the ethanol, and concentrate to obtain a concentrated extract containing the Chinese herbal medicine Bidens pilosa.

[0038] Example 3: The concentrated solutions obtained in Examples 1 and 2 are formulated into oral liquids, granules, capsules, tablets, powder injections, or injections according to the formulation process and technical requirements.

[0039] In this process, starch or magnesium stearate and other excipients are added to the concentrated liquid and mixed. After drying, the mixture is made into tablets by compression or granulation compression. The tablets are then cut into doses and packaged to obtain tablets.

[0040] Add starch or magnesium stearate and other excipients to the concentrated liquid to mix and obtain a soft material. Then, pass it through a 14-16 mesh sieve using a gyratory granulator to obtain wet granules. Place the wet granules in a fluidized bed drying or oven at 60-80℃ to dry them until the moisture content is ≤6.0%. After drying, the granules are granulated by passing them through a 10-12 mesh sieve using a granulator and then divided into doses and encapsulated to make capsules.

[0041] The concentrate was filtered through micropores, freeze-dried, and sterilized to produce a powder injection.

[0042] Distilled water was added to the concentrate, followed by activated carbon. The mixture was heated at 80°C for 30 minutes, coarsely filtered, the pH value was adjusted, and then filtered through a microporous membrane. The solution was then filled and sterilized by high-temperature autoclaving for 30 minutes to obtain the injection solution.

[0043] Example 4: The concentrated solutions obtained in Examples 1 and 2 were used in the preparation of drugs for treating hypertension or for treating complications of hypertension (such as hypertension combined with kidney disease, hypertension caused by kidney disease, and renal hypertension induced by tumor drug treatment).

[0044] Example 5: The concentrated solutions obtained in Examples 1 and 2 were applied to the preparation of blood pressure-lowering foods (such as health foods and functional foods).

[0045] Verification Example 1: Determination of the content of Bidens pilosa extract

[0046] The effective components of the extracts containing the traditional Chinese medicine Bidens pilosa obtained in Examples 1 and 2 were determined by high performance liquid chromatography (HPLC). The results showed that the average content of eight flavonoids—oxcarnitine, luteolin, quercetin, hyperoside, isoquercetin, rutin, luteolin, and quercetin—was 0.08 μg·g. -1 0.36 μg·g -1 1.66 μg·g -1 0.75 μg·g -1 1.65 μg·g -1 0.26 μg·g -1 0.14 μg·g -1 0.05 μg·g -1 This indicates that the total flavonoid content in the extract of Bidens pilosa extracted by this invention is relatively high.

[0047] Verification Example 2: Safety Test of the Active Ingredients in Bidens pilosa Extract

[0048] The safety of the active ingredients in the extracts containing the traditional Chinese medicine Bidens pilosa obtained in Examples 1 and 2 was evaluated using an acute toxicity test in mice. Forty Kunming mice were randomly divided into four groups of ten each. The treatment groups were administered different doses of the active ingredient in Bidens pilosa (2000 mg / kg, 4000 mg / kg, and 8000 mg / kg), while the control group received an equal volume of physiological saline. The drugs were administered by gavage once. The symptoms of poisoning and mortality in the mice were observed within 7 days after administration. The results showed that no obvious symptoms of poisoning or mortality were observed in any group of mice within 7 days after administration. This indicates that the extract of Bidens pilosa obtained in this invention has low acute toxicity and high safety.

[0049] Verification Example 3: Network Pharmacology Test of the Blood Pressure Lowering Mechanism of Active Components from Bidens pilosa

[0050] The potential antihypertensive mechanism of action of the active components of Bidens pilosa was analyzed using network pharmacology. Major active components in Bidens pilosa were screened based on public databases, and their potential targets were predicted; simultaneously, information on disease targets related to hypertension was collected. After mapping analysis, a total of 426 potential common targets were obtained, such as… Figure 1 As shown.

[0051] A network of interactions between Bidens pilosa components and hypertension targets was further constructed, and the network topology parameters were analyzed. The results showed that targets such as TP53, AKT1, and EGFR exhibited high centrality in the network, suggesting they may play a key regulatory role in the intervention of Bidens pilosa on hypertension. Figure 2 and Figure 3 As shown.

[0052] like Figure 4 and Figure 5 As shown, GO functional enrichment and KEGG pathway enrichment analyses were performed on the aforementioned common targets. The results indicate that the active ingredients of Bidens pilosa mainly participate in regulating cellular phosphorylation levels, calcium ion homeostasis, and related signal transduction processes, thereby affecting cellular responses to hormone stimulation and cell motility. KEGG analysis showed that the aforementioned targets were mainly enriched in the PI3K-Akt signaling pathway, calcium signaling pathway, and vascular function protection-related pathways, suggesting that Bidens pilosa may exert a regulatory effect on cardiovascular function through multi-target and multi-pathway synergistic effects.

[0053] Verification Example 4: Test on the antihypertensive mechanism of active ingredients from Bidens pilosa extract against hypertension induced by tumor treatment drugs.

[0054] Specifically, taking the cancer treatment drug anlotinib as an example, this study investigated the antihypertensive mechanism of hypertension induced during anlotinib treatment.

[0055] 1. Experimental Design

[0056] The experiment lasted for 5 weeks, divided into a modeling period and an intervention period. Weeks 1-2 were the modeling period, during which an anlotinib model of hypertension was established via continuous gavage. Weeks 2-5 were the intervention period, during which different doses of Bidens pilosa extract were administered in addition to the continued administration of anlotinib. The experiment ended at the end of week 5, and blood pressure was measured. See Table 1 for details.

[0057] Table 1 Animal Experiment Design and Dosing Schedule

[0058]

[0059] 2. Laboratory animals

[0060] Male SD rats, weighing 200±20 g, were used as experimental animals. There were 6 experimental groups, with 8 rats in each group, for a total of 48 rats, with 2 rats reserved for backup.

[0061] 3. Experimental grouping and dosing regimen

[0062] During the modeling period, except for the normal control group, all other groups were administered anlotinib 1.23 mg / kg / day by gavage daily to induce a hypertension model. The normal control group received an equal volume of normal saline during the same period. The treatment methods for each group during the intervention period were as follows:

[0063] Normal control group: administered physiological saline via gavage throughout the entire process;

[0064] Anlotinib model group: continuous gavage administration of anlotinib;

[0065] Positive control group: In addition to anlotinib, amlodipine 3 mg / kg / d was administered by gavage.

[0066] Bidens pilosa low-dose group: In addition to anlotinib, Bidens pilosa extract 3 g / kg / d was administered by gavage.

[0067] Bidens pilosa medium-dose group: In addition to anlotinib, Bidens pilosa extract 6 g / kg / d was administered by gavage.

[0068] Bidens pilosa high-dose group: In addition to anlotinib, Bidens pilosa extract 9 g / kg / d was administered by gavage.

[0069] Blood pressure was measured in each group of animals before the intervention began. The systolic blood pressure in the anlotinib model group was confirmed to be at least 20 mmHg higher than that in the normal control group, which was considered as a successful model establishment.

[0070] 4. Pharmacodynamic results

[0071] like Figure 6 and Figure 7 As shown, compared with the anlotinib model group, all doses of Bidens pilosa intervention groups could reduce the systolic blood pressure and mean arterial pressure of anlotinib-induced hypertensive rats to varying degrees. The antihypertensive effect was particularly significant in the medium-dose and high-dose groups, and showed a certain dose-dependent trend.

[0072] The results of this experiment show that Bidens pilosa has a clear antihypertensive effect on hypertension induced by tumor drugs, and it is safe and has no other adverse reactions while lowering blood pressure, which is an advantage of traditional Chinese medicine treatment.

[0073] Verification Example 5: Blood Pressure Lowering Test of Bidens pilosa in Hypertensive Patients

[0074] Soak one portion of Bidens pilosa in purified water for 15 minutes, then decoct for 30-45 minutes. Pour out the liquid and store it. Add purified water to cover the herbs again, decoct for another 30 minutes, and combine the decoction with the first decoction. Drink the liquid in several portions as a tea substitute, finishing it within one day. Continue taking this medicine for 3 months, measuring and recording blood pressure daily during the treatment period. See Table 2 for details.

[0075] Table 2. Changes in blood pressure before and after taking Bidens pilosa decoction.

[0076]

[0077] The experimental results in Table 2 show that after taking the drug dosage and method provided by this invention, the patient's blood pressure improved significantly after 3 months. Both diastolic and systolic blood pressure decreased compared to before treatment, indicating that this invention has a clear therapeutic effect in the treatment of hypertension.

[0078] The above descriptions are merely embodiments of the present invention, and common knowledge regarding specific technical solutions or characteristics is not elaborated upon here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the technical solutions of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the implementation of the present invention or the practicality of the patent. The scope of protection claimed in this application should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A method for extracting an extract of a Chinese herbal medicine Bidens pilosa L. characterized by, It comprises the following steps: S1, cut the dry Bidens pilosa whole plant into segments, and soak it in purified water with a mass ratio of 8-10 times of the medicinal material for 1-1.5 hours, then decoct and extract for 1-2 hours, filter out the extract, add 8-10 times of purified water to decoct and extract for 1-2 hours, filter out the extract, combine the two extracts, and concentrate to obtain a concentrated extract; S2, take the concentrated extract obtained in step S1, add ethanol to make the concentration of ethanol reach 60-80%, stand for 24 hours, precipitate, take the supernatant, concentrate to obtain an alcohol precipitation concentrated liquid containing the extract of the Chinese herbal medicine Bidens pilosa.

2. A method for extracting an extract of a Chinese herbal medicine Bidens pilosa L. characterized by, It comprises the following steps: cut the dry Bidens pilosa whole plant into segments, first use 60%-80% ethanol with a mass ratio of 8-10 times of the medicinal material to reflux extract, filter out the extract; then use 60%-80% ethanol with a mass ratio of 8-10 times of the medicinal material to reflux extract, filter out the extract, combine the two filtrates, evaporate ethanol, and concentrate to obtain a concentrated liquid containing the extract of the Chinese herbal medicine Bidens pilosa.

3. The extract obtained by the extraction method of claim 1 or 2.

4. The extraction method of the extract of the Chinese herbal medicine Bidens pilosa according to claim 1 or 2, characterized in that, Add starch or magnesium stearate to the concentrated liquid, mix, dry, and then use a tabletting method or a granulation tabletting method to make tablets, cut into doses, package, and obtain tablets.

5. The extraction method of the extract of the Chinese herbal medicine Bidens pilosa according to claim 1 or 2, characterized in that, Add excipients to the concentrated liquid to obtain a soft material, then sieve the wet granules through a swing granulator to obtain wet granules, dry the wet granules in a boiling drying bed or an oven, sieve the dried granules with a whole granulator, and then fill capsules with the granules to obtain capsules.

6. The extraction method of the extract of the Chinese herbal medicine Bidens pilosa according to claim 5, characterized in that, The excipients are starch or magnesium stearate.

7. The extraction method of the extract of the Chinese herbal medicine Bidens pilosa according to claim 1 or 2, characterized in that, The concentrated liquid is filtered through a microporous filter, freeze-dried, sterilized, and then a powder injection is obtained.

8. The extraction method of the extract of the Chinese herbal medicine Bidens pilosa according to claim 1 or 2, characterized in that, Add distilled water to the concentrated liquid, then add activated carbon, heat at 80°C for 30 minutes, coarse filter, adjust the pH value, filter through a microporous filter membrane, fill, and high-temperature hot-press sterilize for 30 minutes to obtain an injection.

9. The use of the extract obtained by the extraction method of claim 3 in the preparation of a medicine for treating hypertension or a medicine for treating complications of hypertension.

10. The use of the extract obtained by the extraction method of claim 3 in the preparation of a blood pressure-lowering food.

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