CN115770264A - Anti-inflammatory and antioxidant application of hibiscus active ingredient compound - Google Patents

Abstract

The invention discloses anti-inflammation and anti-oxidation uses of hibiscus chrysanthemum active ingredient compounds. At least one of hibiscus active ingredient compounds containing wormwood and kaempferol or pharmaceutically acceptable salts and stereoisomers thereof is used as an active ingredient compound for preparing anti-inflammatory and anti-oxidative medicines. The hibiscus chrysanthemum active ingredient compound provided by the invention can be effectively used in the preparation of anti-inflammatory and antioxidant drugs, and provides a new approach for the selection of anti-inflammatory and antioxidant drugs.

Description

Anti-inflammation and anti-oxidation application of hibiscus chrysanthemum active ingredient compound

technical field

The invention belongs to the technical field of medicine, and in particular relates to the anti-inflammation and anti-oxidation application of hibiscus chrysanthemum active ingredient compounds.

Background technique

Hibiscus chrysanthemum is a plant of the Asteraceae genus Crossostephium Less. It is mainly distributed in the south of the Yangtze River in my country. Hibiscus chrysanthemum is a rare "white leaf plant", so it is widely used as a landscape plant to decorate homes and urban greening. Hibiscus chrysanthemum is widely cultivated in South China for appreciation and medicinal purposes. One of the main subshrub vegetation in Dachen Island, southeast of Jiaojiang River, Taizhou City, Zhejiang Province is Hibiscus chrysanthemum. The leaves of hibiscus chrysanthemum were used as medicine and were first recorded in "Compendium of Materia Medica". The root is used as medicine to treat rheumatic joint pain and epigastric cold pain. Folks often use hibiscus chrysanthemum as a herbal medicine for treating convulsions in children, and it also has the effect of relieving acne and itching; in Guangdong folks, the whole plant is used to treat diabetes. Hibiscus chrysanthemum has relatively abundant plant resources and has the potential to be developed into medicine.

Hibiscus contains chemical components such as volatile oils, flavonoids, terpenoids and coumarins. Artesin is one of the main sesquiterpenoids in hibiscus. Artesin was first obtained from Artemisia santolina Modern researches have proved that artemisinin has certain anti-trichomonas activity, has non-competitive inhibitory effect on acetylcholine-induced frog abdominal muscle contraction, and presents a dose-dependent effect, at 2×10 ^-4 mol/L The degree of inhibition was 74%. However, there is no relevant report about the anti-inflammatory and anti-oxidative use of hibiscus chrysanthemum active ingredient compounds.

Contents of the invention

Aiming at the above-mentioned deficiencies in the prior art, the present invention provides an anti-inflammation and anti-oxidation application of hibiscus chrysanthemum active ingredient compounds.

In order to achieve the above-mentioned purpose of the invention, the technical scheme adopted in the present invention is:

Provides an anti-inflammatory and anti-oxidative application of active ingredient compounds of hibiscus chrysanthemums, using at least one of the active ingredient compounds of hibiscus chrysanthemums containing grass wormwood and kaempferol or their pharmaceutically acceptable salts and stereoisomers as the active ingredient The component compounds are used to prepare anti-inflammatory and anti-oxidative medicines.

Further, the hibiscus chrysanthemum active ingredient compound containing wormwood and kaempferol is extracted and separated from the hibiscus chrysanthemum plant of the genus Artemisia genus in the family Asteraceae.

Further, the method for extracting and separating hibiscus active ingredient compounds containing wormwood and kaempferol from hibiscus includes the following steps:

S1. After natural drying, the whole hibiscus chrysanthemum was crushed with a pulverizer, passed through a 20-mesh sieve, and extracted twice with chloroform or dichloromethane at a weight-to-volume ratio of 1:20, 1 hour each time, and combined extracting liquid, recovering the solvent under reduced pressure to obtain extract, and dispersing the extract evenly in distilled water to obtain suspension;

S2. The suspension is subjected to silica gel column chromatography, and gradient elution is performed with cyclohexane-ethyl acetate, petroleum ether-acetone, chloroform-ethyl acetate, dichloromethane-ethyl acetate, and the eluent is received by fractions , collect all fractions containing wormwood and kaempferol, concentrate under reduced pressure, and recrystallize with acetone to obtain hibiscus active ingredient compounds containing wormwood and kaempferol.

The dosage form of the hibiscus chrysanthemum active ingredient compound of the present invention is not limited, as long as it is a dosage form that can make the active ingredient effectively reach the body, such as tablet, capsule, powder, granule, syrup, solution, suspension, injection, tincture , oral liquid, aerosol, buccal, granules, pills, powder and other common dosage forms or sustained-release dosage forms such as nano-preparations.

In addition to the main active ingredient, it may also contain a small amount of secondary ingredients and/or pharmaceutically acceptable carriers that do not affect the active ingredient, for example: may contain sweeteners to improve taste, antioxidants to prevent oxidation, and Necessary excipients for various preparations, etc.

The beneficial effects of the present invention are:

The hibiscus chrysanthemum active ingredient compound provided by the invention can be effectively used in the preparation of anti-inflammatory and antioxidant drugs, and provides a new approach for the selection of anti-inflammatory and antioxidant drugs.

Detailed ways

The specific embodiments of the present invention are described below so that those skilled in the art can understand the present invention, but it should be clear that the present invention is not limited to the scope of the specific embodiments. For those of ordinary skill in the art, as long as various changes Within the spirit and scope of the present invention defined and determined by the appended claims, these changes are obvious, and all inventions and creations using the concept of the present invention are included in the protection list.

Example 1

Extraction and separation of hibiscus chrysanthemum active ingredient compounds:

S1. Take 1 kg of hibiscus chrysanthemum whole herb, dry it naturally, crush it with a pulverizer, pass through a 20-mesh sieve, and extract it twice with 20 L of chloroform, 1 hour each time, combine the extracts, and recover the solvent under reduced pressure to obtain an extract , uniformly disperse the extract into distilled water to obtain a suspension;

S2. The suspension was subjected to silica gel column chromatography, with a gradient of cyclohexane-ethyl acetate, petroleum ether-acetone, chloroform-ethyl acetate, dichloromethane-ethyl acetate (10:0→9:1→ 4:1 → 1:1 → 0:10), receive the eluate according to fractions, collect all fractions containing wormwood and kaempferol, concentrate under reduced pressure, and recrystallize with acetone to obtain The hibiscus chrysanthemum active ingredient compound of kaempferol has an HPLC purity greater than 99.8%.

Example 2

Preparation of hibiscus chrysanthemum active ingredient compound tablet:

In this example, the hibiscus chrysanthemum active ingredient compound containing wormwood and kaempferol extracted and separated in Example 1 was used to prepare tablets. Purpose coarse powder, then the coarse powder is mixed as the main raw material with the corresponding auxiliary materials (Baihujing: lactose=7:3, mass ratio), granulated after mixing, added sodium stearate and mixed evenly, and then compressed into tablets to make Tablets containing active ingredient compounds of the hibiscus family.

Example 3

Preparation of hibiscus chrysanthemum active ingredient compound powder injection:

In this example, the hibiscus chrysanthemum active ingredient compound containing wormwood and kaempferol extracted and isolated in Example 1 was used to prepare a powder injection, and the hibiscus chrysanthemum active ingredient compound and mannitol were dissolved in water for injection, and the volume was constant. Filter the obtained solution, put it into vials, 1mL per bottle, freeze-dry, seal and sterilize, and obtain each freeze-dried powder injection containing the active ingredient compound of hibiscus chrysanthemum.

Example 4

Preparation of hibiscus chrysanthemum active ingredient compound capsules:

In this embodiment, the hibiscus chrysanthemum active ingredient compound containing wormwood and kaempferol extracted and separated in Example 1 is used to prepare capsules. First, the hibiscus chrysanthemum active ingredient compound is vacuum-dried until the water content is 6%, and then crushed into 20 Purpose coarse powder, then the coarse powder is used as the main raw material and the corresponding auxiliary material (Baihujing: lactose=7:3, mass ratio) is mixed, after mixing, carry out granulation, after granulation, pack in the capsule, make corresponding containing Capsules of hibiscus active ingredient compounds.

Example 5

Anti-inflammatory experiment of hibiscus chrysanthemum active ingredient compounds:

This embodiment adopts 40 SPF grade healthy Wistar rats, body weight (230 ± 10) g, after the rats are fed adaptively for 1 week, 10 are randomly selected as a control group, and 10 are low-dose groups, and the administration is 1.0g/ kg/d dose of hibiscus chrysanthemum active ingredient compounds; 10 for the middle dose group, administered 2.0g/kg/d dose of hibiscus chrysanthemum active ingredient compounds; 10 for the high dose group, administered 4.0g/kg/ The active ingredient compound of hibiscus chrysanthemum in dose d; the product prepared in Example 1 was selected as the active ingredient compound of hibiscus chrysanthemum. The same volume of distilled water was administered to the control group, and the administration continued for 7 days in each group. On the 8th day, sodium urate was injected into the ankle joints of the rats. According to the concentration of 0.25g/10ml, each rat was injected with 0.2ml, and the rats were killed after 48h. Whole blood was collected, centrifuged at 3000r/min for 15min after standing for 2h, and the supernatant was taken for use. Elisa method was used to detect interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-1β (IL-1β), C-reactive protein (CRP).

Table 1 The effect of hibiscus chrysanthemum active ingredient compounds on 5 inflammatory factors in rats with acute inflammation

group IL-6(mg/L) IL-8(mg/L) IL-1β(mg/L) CRP (mg/L) control group 18.56 24.87 9.98 3.25 low dose group 17.06 22.98 9.02 3.01 Middle dose group 15.33 20.36 8.56 2.56 high dose group 12.36 18.96 7.36 2.02

The present invention obtains the following main conclusion through the anti-inflammatory experiment of hibiscus chrysanthemum active ingredient compounds: the hibiscus chrysanthemum active ingredient compound of the present invention can reduce the content of inflammatory factors CRP, IL-1β, IL-6 and IL-8 in animals with acute inflammation. This example shows that the active ingredient compound of hibiscus chrysanthemum has good anti-inflammatory activity, and can be used as an anti-inflammatory drug to prevent or treat inflammation caused by high levels of inflammatory factors CRP, IL-1β, IL-6, and IL-8.

Example 5

Antioxidant test of hibiscus chrysanthemum active ingredient compounds:

In order to evaluate the scavenging ability of the hibiscus chrysanthemum active ingredient compound of the present invention to oxidative free radicals, the product extracted and separated in Example 1 was used to test its ability to scavenge free radicals by DPPH free radical scavenging experiment.

In ethanol solution, DPPH can form purple stable free radicals with strong absorption at 517nm, and free radical scavengers can make it fade. If the hibiscus chrysanthemum active ingredient compound in the present invention can effectively scavenge DPPH, it means that it can effectively reduce the concentration of various free radicals and block the peroxidation reaction.

Prepare 10 mg/mL hibiscus chrysanthemum active ingredient compounds in Example 1 with absolute ethanol, and then dilute to sample solutions with concentrations of 8, 6, 4, 2, and 1 mg/mL, which are designated as samples 1-5. Take 100 μL of 0.1 mmol/L DPPH solution, add 100 μL of sample solution, mix and react in the dark at room temperature for 30 minutes, measure the absorbance (A _i ) at 517 nm, parallel 3 times, and take the effective average value. At the same time, mix 100 μL of DPPH solution with 100 μL of absolute ethanol, and measure its absorbance (A ₀ ) according to the above method. Mix 100 μL of sample solution with 100 μL of absolute ethanol, and measure its absorbance (A _j ) according to the above method.

DPPH clearance rate (%)=[1-(A _i -A _j )/A ₀ ]×100%

Table 2 sample 1-5DPPH free radical scavenging experiment test result

serial number sample 1 sample 2 sample 3 Sample 4 Sample 5 DPPH clearance rate (%) 70.56 65.36 60.22 55.98 48.32

It can be seen from the above table that the active ingredient compound of hibiscus chrysanthemum of the present invention has good free radical scavenging effect. It can help skin cells resist free radical stimulation and maintain the normal function of cells.

The hibiscus chrysanthemum active ingredient compound provided by the invention can be effectively used in the preparation of anti-inflammatory and antioxidant drugs, and provides a new approach for the selection of anti-inflammatory and antioxidant drugs.

It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, but that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Accordingly, the embodiments should be regarded in all points of view as exemplary and not restrictive, the scope of the invention being defined by the appended claims rather than the foregoing description, and it is therefore intended that the scope of the invention be defined by the appended claims rather than by the foregoing description. All changes within the meaning and range of equivalents of the elements are embraced in the present invention.

In addition, it should be understood that although this specification is described according to implementation modes, not each implementation mode only contains an independent technical solution, and this description in the specification is only for clarity, and those skilled in the art should take the specification as a whole , the technical solutions in the various embodiments can also be properly combined to form other implementations that can be understood by those skilled in the art.

Claims (3)

1. An anti-inflammatory and antioxidant use of hibiscus active ingredient compounds is characterized in that the hibiscus active ingredient compounds containing artemisinin and kaempferol or at least one of pharmaceutically acceptable salts and stereoisomers of the hibiscus active ingredient compounds are used as the active ingredient compounds for preparing anti-inflammatory and antioxidant medicines.

2. The anti-inflammatory and antioxidant use of the active ingredient compounds of hibiscus according to claim 1, wherein the active ingredient compounds of hibiscus comprising artemisinin and kaempferol are extracted and separated from hibiscus of mugwort of Compositae.

3. The anti-inflammatory and antioxidant use of hibiscus cannabinus active ingredient compounds according to claim 2, wherein the method for extracting and separating the hibiscus cannabinus active ingredient compounds containing the artemisinin and the kaempferol from hibiscus cannabinus comprises the following steps:

s1, naturally airing the whole plant of hibiscus mutabilis, crushing the whole plant of hibiscus mutabilis by using a crusher, sieving the crushed plant by using a 20-mesh sieve, and mixing the crushed plant with chloroform or dichloromethane according to a weight-volume ratio of 1:20, reflux-extracting for 2 times, each time for 1h, mixing the extracting solutions, recovering the solvent under reduced pressure to obtain an extract, and uniformly dispersing the extract into distilled water to obtain a suspension;

and S2, subjecting the suspension to silica gel column chromatography, performing gradient elution by using cyclohexane-ethyl acetate, petroleum ether-acetone, trichloromethane-ethyl acetate and dichloromethane-ethyl acetate, receiving eluent according to fractions, collecting all fractions containing the artemisinin and the kaempferol, performing reduced pressure concentration, and performing recrystallization by using acetone to obtain the hibiscus active ingredient compounds containing the artemisinin and the kaempferol.