HK1081115B - Extraction and purification method of active constituents from stem of lonicera japonica thunb., its usage for anti-inflammatory and analgesic drug - Google Patents

Description

Method for extracting and purifying active ingredients from honeysuckle stem, and application of active ingredients in anti-inflammatory and analgesic drugs

Technical Field

The present invention relates to a method for extracting and purifying active ingredients from honeysuckle (Lonicera japonica Thunb) and applications thereof. More particularly, the present invention relates to a method for extracting and purifying active ingredients including swertiamarin from honeysuckle stem (leaf-removed honeysuckle stem) by removing tannin, poorly soluble flavonoid, saponin, etc. The active ingredient thus obtained has better anti-inflammatory and analgesic effects, is safer and more stable than the conventional active ingredients obtained from honeysuckle flower or honeysuckle leaf, and includes swertiamarin, which is an effective active ingredient of anti-inflammatory and analgesic drugs.

Background

Lonicera japonica (Lonicera japonica) is a semievergreen ligand rattan shrub that naturally grows under mountains or levee feet at elevations ranging from 50-600 meters in Japan, China, and Korea. The flower bud (honeysuckle flower) and stem (honeysuckle stem) are used as herbal medicine for promoting urination, removing toxic substance, stopping bleeding, purifying blood, treating tumor, and treating edema, common cold, diarrhea, emesis, etc. [ Korean plant textbook interpretation, Chang-Bok Lee, 709, 1989, Hangmun Publishing Co., Seoul; herbal standards Not included In the Korean Pharmacopoeia (Standard for Herbs (Herb Medicines) Not Coveredby Korean Pharmacopeia), Hyung-Joon Chi, Sangg-In Lee, 87, 305, 1988, Korea Medical Index Co., Seoul; korean plant resources (ResourcePlants in Korea), Tae-Kyung Kim, vol.4, 148-149, 1996, SNU Press, Seoul. In addition, many traditional chinese Medicine books, including the Sasang connective Medicine and Gwangje Bigeup, teach that it can treat a wide variety of inflammatory abscesses both inside and outside the human body [ Sasang connective Medicine for Chosing Pen, Yonben Chosing Medical Institute, 276, 1991, Yodogg publishing Co., Seoul; gwangje Bigeup, Kyung-Hwa Lee, 349-. Because of its anti-inflammatory analgesic activity, has long been used as a folk medicine for treating upper respiratory tract infections such as cold, tonsillitis and neuralgia. Recently, the anti-inflammatory and analgesic activity of lonicera japonica has been confirmed through various experimental animal models and its effective physiologically active ingredient has been isolated and reported in academic scope [ "development of plant anti-inflammatory drugs: comparison of anti-inflammatory and analgesic effects of extracts of Lonicera japonica Thunb "(Development of the novel antibiotic and analgesic media: Complex of anti-antibiotic and analgesic activities of the homeopathic extract), Song-Jin Lee et al, Korean Journal of pharmacy, 363-367, 25, 1994; "Flavonoids from the aerial parts of Lonicera japonica" (Flavonoids from the ear of Lonicera japonica), Son et al, proceedings of the research on drugs (Arch. pharm. Res.), 365-; "anti-inflammatory Activity of Lonicera japonica" (anti inflammatory activity of Lonicera japonica), Lee et al, phytotherapy research (Phyto. Res.), 445-447, 12, 1998; "Triterpenoid saponins from the terrestrial part of Lonicera japonica" (Triterpenoid saponins from the botanical parts of Loiiiceia japonica), Son et al, phytochemistry, 1005-, 1008, 35, 1994; "Anti-inflammatory activity of the major component of honeysuckle" (Anti-inflammatory activity of the major constituents of Lonicera japonica), Lee et al, "journal of the pharmaceutical research (Arch. pharm. Res.), 133-135, 18, 1995).

Description of the related Art

Heretofore, hydrolyzable tannins such as chlorogenic acid, methyl caffeate, chlorogenic acid and isochlorogenic acid and iridoid glycosides such as loganin, sweroside, vogeloside and epi-vogeloside have been called effective active ingredients contained in honeysuckle stem. Most of the conventional research has focused on honeysuckle flowers and honeysuckle leaves. It should be noted that the honeysuckle stem has a different composition distribution than the honeysuckle leaves or flowers. That is, unlike the stem of honeysuckle, the main components of honeysuckle leaves or honeysuckle flowers are: flavonoids such as loniceraside, rhoifolin, and alkanna tinctoria flavone; triterpene saponin with hederagenin or oleanolic acid as non-sugar component; and various hydrolyzable tannins.

It is difficult to prepare these components into injections. If the injections are rich in polymer tannins, they can combine with other components to produce a coprecipitation and can combine with serum proteins in blood to form a poorly soluble precipitate, which may be the cause of vascular stenosis. Furthermore, since flavonoids included in honeysuckle are generally insoluble in water, it is essential to dissolve them to a level higher than an effective concentration with a considerable amount of organic solvent or other agent as a dissolution mediator. In addition, flavonoid-rich active ingredients are extremely poorly soluble in physiological saline solutions for injection and may become unstable if stored in alkaline buffer solutions for a long period of time. Finally, saponins, in particular mono-saponins (monodesmosides), obtained from honeysuckle are known to have a strong hemolytic effect. Therefore, they cannot be injected directly into the vein without purification [ "study on saponin" (Studies on the saponin of Lonicera japonica Thunb.), Kawai et al, J.P.Pharmacol. (chem. pharm. Bull.), 4769-4775, 36(12), 1988 ]. The tannin and insoluble flavonoid contents of the honeysuckle leaves and the honeysuckle flowers are higher than those of the honeysuckle stems. They also produce more toxicity at low levels and provide less pain relief and anti-inflammatory effects in acute toxicity tests for injection.

Lonicera japonica, swertia japonica, gentian, gentiana triflora, gentiana yedoensis, gentiana rigescens have been used for many years to alleviate fever or to detoxify. However, it is not clearly understood which components provide such effects and most studies have focused on identifying the activity of the main pharmaceutical ingredient loganin of honeysuckle stem [ journal of natural products (j. nat. prod.), 54(4), 1102-: botanical drug (Planta Med.), 60, 232, 234, 1994: study of plant therapy (Phytotherapy Res.), 12, 405, 408, 1998.

Furthermore, it is considered that the drug glycosides only protect the liver effectively and inhibit the bacterial activity [ journal of national pharmacology (Ethnopharmacol) ], 42, 183-191, 1994: brief introduction to medicinal chemistry (chem. pharm. Bull.), 45(11), 1823-1827, 1997: yakugaku Zasshi, 102(8), 755-.

Disclosure of Invention

The present inventors have determined that an active ingredient of honeysuckle stem from which tannin, poorly soluble flavonoid, saponin, etc. are removed (honeysuckle stem from which leaves are removed) and an effective active ingredient swertioside among the active ingredients have excellent anti-inflammatory and analgesic effects.

Accordingly, it is an object of the present invention to provide a method for preparing an active ingredient having excellent anti-inflammatory and analgesic activities, safety and stability from honeysuckle stem.

It is another object of the present invention to provide anti-inflammatory and analgesic agents comprising said active ingredients.

It is another object of the present invention to provide anti-inflammatory and analgesic agents that include swertin.

Brief Description of Drawings

FIG. 1 is a schematic diagram showing the pharmaceutical effect of active ingredients with varying amounts.

Description of The Preferred Embodiment

The present invention provides a method for preparing an active ingredient having excellent anti-inflammatory and analgesic activities, safety and stability from honeysuckle stem.

The invention also provides anti-inflammatory and analgesic agents comprising said active ingredients.

The invention also provides an anti-inflammatory and analgesic drug comprising said swertiamarin.

The following gives a detailed description of the present invention.

The present invention relates to a method for extracting and purifying active ingredients and swertiamarin from honeysuckle by removing tannin, poorly soluble flavonoid, saponin, etc. The thus obtained active ingredient has better anti-inflammatory and analgesic effects, is safer and more stable than the conventional active ingredients obtained from honeysuckle flower or honeysuckle leaf, and includes swertiamarin, which is an effective active ingredient of anti-inflammatory and analgesic drugs. Active ingredients and swertiamarin were extracted from honeysuckle stem and purified by the following method.

The honeysuckle stem sample was extracted under reflux with about 7-10 volumes of distilled water for 2-3 hours and then filtered. The residue was collected and extracted with about 5-7 volumes of distilled water under reflux for 2-3 hours. The liquid thus obtained was filtered and combined with the above filtrate, concentrated under reduced pressure and filtered again so that the volume thereof was about 1 to 3 times (v/w) the weight of the herbal medicine. In the course of extraction with distilled water, if too little distilled water is used, stirring becomes difficult and extraction efficiency is lowered because the solubility of the extract becomes poor. On the contrary, if an excessive amount of distilled water is used, the time and cost required are excessively high. Thus, an equal amount of water saturated lower alcohol is added and stirred at 30-50rpm for about 10-20 minutes. After layer separation, the water-saturated lower alcohol layer was filtered and concentrated under reduced pressure to obtain the primary active ingredient. Herein, water-saturated lower alcohols are prepared by adding distilled water to lower alcohols such as propanol and butanol with stirring, followed by settling. The layer separation process was carried out 2-3 times. In the process of obtaining the lower alcohol solvent fraction, if too little lower alcohol is used, the purification efficiency is lowered, whereby the extraction yield and the content of effective ingredients are lowered. On the contrary, if too much lower alcohol is used, the cost is increased. Therefore, it is recommended to use lower alcohol in a volume (v/w) of 1 to 3 times the weight of the herb.

The primary active ingredient is subjected to column chromatography using polyamide resin, polyvinylpyrrolidone resin or the like to remove unnecessary substances and detect the effective ingredient. The filler is used in 1-10 volumes (w/w) of a layer of water-saturated lower alcohol. Eluting 2-3 volumes of 50% (v/v) methanol and methanol with the packing volume and then eluting the distilled aqueous solvent by a step gradient method. The secondary active ingredient obtained by eluting the active ingredient with distilled water contains less aromatic organic acids, tannins and flavonoids and exhibits better medicinal effects, significantly reduced toxicity, increased solubility and improved blood stability. The secondary active ingredient was subjected to further column chromatography using (octadecylsilane) resin. Starting from 10% (v/v) methanol, 2-3 volumes of solvent resin were eluted by a step gradient method while increasing the methanol content by 10% (v/v). Polyamide or polyvinylpyrrolidone resins are used in a volume 20-50 times the weight of the purified primary active ingredient. The active ingredient obtained by eluting 20-30% (v/v) methanol showed the best anti-inflammatory and analgesic activity. Analysis of these active ingredients revealed that iridoid substances such as swertin and loganin are major active ingredients. Contains swertiamarin 15.1-72.1 wt% and loganin 13.9-41.4 wt%.

The active ingredient obtained by eluting 20-30% (v/v) methanol has the highest swertiamarin content. Then subjecting the active ingredients to column chromatography to isolate swertiamarin represented by the following chemical formula 1:

the thus obtained active ingredient and swertiamarin were subjected to an arachidonic acid-induced ear edema test and a croton oil-induced ear edema test to determine anti-inflammatory effects. And an acetic acid induced writhing test was performed to determine analgesic effect. As a result, they were found to have anti-inflammatory and analgesic activities far superior to those of the conventional active ingredients obtained from honeysuckle flower or honeysuckle leaf.

The swertiamarin obtained by the present invention can be prepared into a therapeutic agent by a method well known in the pharmaceutical field. It can also be administered orally or parenterally, alone or together with pharmaceutically acceptable carriers, shaping agents, diluents, and the like. It can be made into powder, granule, tablet, capsule, syrup, skin ointment or injectable medicine.

The human dose of the active ingredient or swertiamarin of the present invention can be selected according to the absorption, inactivation rate and excretion rate of the active ingredient in the body, age, sex and physical condition of the subject, severity of the disease to be treated, etc. Preferably, 1-200mg of the active ingredient or swertiamarin is administered to an adult. Administration is according to the prescribed method and, if necessary, is decided according to the expert's advice. The drug may be administered several times per day at regular intervals, preferably 1-3 times per day. The pharmaceutical composition may be administered orally or non-orally. When administered parenterally, it may be administered intravenously, intramuscularly, rectally, or transdermally.

Because the active ingredients of honeysuckle stem have excellent anti-inflammatory and analgesic effects and ideal solubility, acute toxicity and blood stability, they are very suitable for injectable drugs.

The present invention is described more specifically by the following examples. However, the following examples are only for the understanding of the present invention and they should not be construed as limiting the scope of the present invention.

Examples

Example 1: medicinal action of every part of honeysuckle

Samples of whole plants of Lonicera japonica (Lonicera japonica with stems and leaves), Lonicera japonica leaves, and Lonicera japonica stems were collected at 7 months in 1999 in Yeong cheon, Gyeong sangbuk-do in Korea. The sample was dried in the shade and extracted with 7 volumes of distilled water under reflux for 2.5 hours and then filtered. The residue was then collected and extracted with 7 volumes of distilled water under reflux for 2.5 hours. The liquid thus obtained was filtered and combined with the above filtrate, concentrated under reduced pressure and filtered again so that the volume thereof was about 2 times (v/w) the weight of the herbal medicine. An equal volume of water saturated n-butanol was then added and the mixture was stirred at about 30rpm for 15 minutes. After layer separation, the alcohol layer was filtered and concentrated under reduced pressure to obtain the primary active ingredient. Column chromatography was then performed using polyamide resin (CAS NO.63428-83-1) to obtain a purified fraction. The amount of resin was 5 volumes of sample. The secondary active ingredient was obtained by eluting 2 volumes of 50% (v/v) methanol and then eluting the distilled aqueous solvent by a step gradient method. Croton oil-induced ear edema test was performed by administering the secondary active ingredients of the whole honeysuckle, stem and leaves to the tail vein of 6-week-old ICR mice (body weight: 20-30g, n-6, SLC, japan) that had been fasted for 4 hours. After 15 minutes, inflammation was induced with 2.5% croton oil. After 4 hours, the thickness of the left and right mouse ears was measured using a calibrated thickness gauge. The inflammation rate was calculated by the following equation 1 and the results are shown in table 1.

Equation 1

Inflammation rate (%) [ thickness of inflamed (right) ear-thickness of normal (left) ear ]/[ thickness of normal ear ] X100

TABLE 1

As shown in table 1, the active ingredients of the honeysuckle stem showed the best anti-inflammatory and analgesic activity.

Example 2: comparison of pharmaceutical action of active ingredient obtained from honeysuckle stem

Croton oil-induced ear edema test was performed as described in example 1. Furthermore, arachidonic acid-induced ear edema test was performed by administering drugs (marobiven, a primary component of honeysuckle stem and a secondary active component of honeysuckle stem) to the caudal vein of 6-week-old ICR mice (body weight: 20-30g, n-6, SLC, japan) that had been fasted for 4 hours. After 15 minutes, inflammation was induced with 0.05% arachidonic acid. After about 1 hour, the thickness of the left and right ears of the mouse was measured and the inhibition rate was calculated by equation 1 and the results are shown in table 2.

TABLE 2

As shown in table 2, the secondary active ingredient obtained from the honeysuckle stem does not include such compounds as aromatic organic acids, tannins and flavonoids and has an active ingredient content higher than that of the primary active ingredient.

Example 3: comparison of pharmaceutical action of active ingredient obtained from honeysuckle stem

The procedure of example 1 was carried out by substituting polyvinylpyrrolidone resin (CAS NO.25249-54-1) for the polyamide.

TABLE 3

Example 4: preparation of the final active ingredient and drug action test

The secondary active ingredient of the honeysuckle stem prepared in example 1 was concentrated under reduced pressure to obtain powder. The powder was subjected to column chromatography again using ODS resin (YMC GEL ODS-A12 nm, S-150m or ODS-AM 12nm, S-50m or ODS-AQ 12nm, S-50 m). The final active ingredient was obtained using 3 volumes of resin and eluting 20% (v/v) methanol.

Croton oil-induced ear edema test was performed as described in example 1.

In addition, acetic acid-induced writhing test was performed by administering the drug (marobiven, and the last active ingredient of honeysuckle stem) to the tail vein of ICR mice (body weight: 20-30g, n-8, SLC, japan) that had been fasted for 1 day. After 20 minutes, 0.7% acetic acid was injected intraperitoneally. After 15 minutes, the number of writhing was counted for 10 minutes to calculate the inflammation inhibition rate. The results are shown in table 4.

TABLE 4

Example 5: active ingredient content and drug action test

The active ingredient content was identified from High Performance Liquid Chromatography (HPLC) performed on the final active ingredients prepared in examples 1 and 4: swertisin 15.1-72.1 wt% and strychnine 13.9-41.4 wt%. The swertiamarin and loganin contents of each sample are shown in table 5. The pharmaceutical effect of the final active ingredient varying with the place of production and the time of collection is shown in table 6.

Table 5: the contents of swertiamarin and loganin vary with the production area and collection time

Moon cake	Yeongcheon，Gyeongsangbuk-do(A)	Andong，Gyeongsangbuk-do(B)	Taishan, China	New countryside, China
					1 month	Lo: 34.6 Sw: 57.7 in total: 92.3	Lo: 38.3 Sw: 55.4 in total: 93.7	Lo: 38.2 Sw: 48.6 in total: 86.8	Lo: 31.4 Sw: 59.2 in total: 90.6
2 month	Lo: 30.2 Sw: 64.1 in total: 94.3	Lo: 32.3 Sw: 62.1 in total: 94.4	Lo: 33.3 Sw: 54.2 in total: 87.5	Lo: 28.6 Sw: 63.3 in total: 91.9
					3 month	Lo: 22.7 Sw: 70.2 in total: 92.9	Lo: 26.1 Sw: 67.6 Total: 93.7	Lo: 25.8 Sw: 60.3 Total: 86.1	Lo: 20.5 Sw: 72.1 in total: 92.6
4 month	Lo: 13.9 Sw: 63.1 in total: 77.0	Lo: 15.9 Sw: 60.1 in total: 76.0	Lo: 19.1 Sw: 65.3 Total: 84.4	Lo: 14.2 Sw: 69.7 in total: 83.9
					Month 5	Lo: 17.1 Sw: 52.3 in total: 69.4	Lo: 14.2 Sw: 55.7 in total: 69.9	Lo: 18.7 Sw: 66.2 in total: 84.9	Lo: 14.9 Sw: 55.2 in total: 70.1
6 month	Lo: 19.4 Sw: 33.4 in total: 52.8	Lo: 16.2 Sw: 38.2 in total: 54.4	Lo: 23.5 Sw: 43.5 Total: 67.0	Lo: 17.4 Sw: 42.6 in total: 60.0
					7 month	Lo: 21.7 Sw: 15.1 in total: 36.8	Lo: 25.3 Sw: 17.5 in total: 42.8	Lo: 28.9 Sw: 29.8 in total: 58.7	Lo: 22.4 Sw: 35.2 in total: 57.6
8 month	Lo: 23.5 Sw: 29.4 in total: 52.9	Lo: 27.0 Sw: 32.7 in total: 59.7	Lo: 30.1 Sw: 35.9 in total: 66.0	Lo: 32.1 Sw: 36.4 in total: 68.5
					9 month	Lo: 25.3 Sw: 47.6 Total: 72.9	Lo: 30.1 Sw: 45.3 in total: 75.4	Lo: 33.6 Sw: 42.5 Total: 76.1	Lo：39.2 Sw: 40.2 Total: 79.4
10 month	Lo: 28.7 Sw: 52.2 in total: 80.9	Lo: 32.2 Sw: 50.5 Total: 82.7	Lo: 33.8 Sw: 48.2 in total: 82.0	Lo: 37.4 Sw: 43.3 Total: 80.7
					11 month	Lo: 39.6 Sw: 54.3 in total: 93.9	Lo: 37.3 Sw: 58.3 in total: 95.6	Lo: 38.0 Sw: 54.6 in total: 92.6	Lo: 41.4 Sw: 45.6 in total: 87.0
12 month	Lo: 36.2 Sw: 56.8 in total: 93.0	Lo: 33.7 Sw: 60.2 Total: 93.9	Lo: 35.2 Sw: 58.3 in total: 93.5	Lo: 40.2 Sw: 52.6 in total: 92.8

Lo: loganin; sw: swertia herb glycoside

TABLE 6

Experimental results for croton oil-induced ear edema

Results of acetic acid induced writhing test

Example 6: hemolysis assay of the final active ingredient

20mL of blood was collected from the rabbit heart using a syringe treated with heparin. The blood was centrifuged for about 10 minutes. The supernatant was decanted and the residue was diluted with 10 volumes of physiological saline solution for injection. After mixing by slow shaking, 0.5mL of diluted blood and 0.5mL of each of the drugs of example 1 (the final active ingredient of honeysuckle stem and the primary active ingredients of honeysuckle stem, honeysuckle leaves and whole honeysuckle) and the final active ingredient of example 4 were put into a test tube (a physiological saline solution and a distilled water (100% hemolysis) control group were also prepared). The test tubes were kept warm in a bath maintained at 37 ℃ for 15 minutes and then left at room temperature for 45 minutes. Finally after centrifugation at 2500rpm for 2 minutes, the upper layer was analyzed at 540 nm.

TABLE 7

Categories	Hemolysis (%)	Standard deviation of
			Control group (physiological saline solution for injection)	0.73	0.17
Final active ingredient, 5X 10g/ml	0.69	0.10
			Final active ingredient, 1.5X 10g/ml	0.73	0.11
Final active ingredient, 5X 10g/ml	0.74	0.88
			Honeysuckle stem, 5X 10 at bestg/ml	15.2	0.20
Lonicera japonica leaf, 5X 10 in maximumg/ml	35.1	0.15
			Lonicera japonica Thunb, 5X 10 in maximumg/ml	23.0	0.45

Example 7: preparation of swertiamarin

The final active ingredient prepared in example 4 was subjected to column chromatography using octadecylsilane resin to isolate swertiamarin represented by the following chemical formula 1:

example 8: determination of swertiamarin anti-inflammatory action

Croton oil-induced ear edema test was performed by administering swertia pseudochinensis to the tail vein of 6-week-old ICR mice (body weight: 20-30g, n-6, SLC, japan) that had been fasted for 4 hours. After 15 minutes, inflammation was induced with 2.5% croton oil. After 4 hours, the thickness of the left and right mouse ears was measured using a calibrated thickness gauge. The inflammation rate was calculated by equation 1 and the results are shown in table 8.

The effect of orally administered drugs is shown in table 9.

TABLE 8

TABLE 9

Furthermore, arachidonic acid-induced ear edema test was performed by administering swertin to the tail vein of 6-week-old ICR mice (body weight: 20-30g, n: 6, SLC, japan) that had been fasted for 4 hours. After 15 minutes, inflammation was induced with 0.05% arachidonic acid. After 1 hour, the thickness of the left and right ears of the mouse was measured and the inhibition rate was calculated by equation 1 and the results are shown in table 10. The effect of orally administered drugs is shown in table 9.

Watch 10

TABLE 11

Example 9: determination of analgesic Effect of swertiamarin

Acetic acid-induced writhing test was performed by administering swertisin to the caudal vein of ICR mice (body weight: 20-30g, n-8, SLC, japan) that had been fasted for 1 day. After 20 minutes, 0.7% acetic acid was injected intraperitoneally. After 15 minutes, the number of writhing was counted for 10 minutes to calculate the inflammation inhibition rate. The results are shown in table 12. The effect of orally administered drugs is shown in table 13.

TABLE 12

Watch 13

Example 10: toxicity test

The honeysuckle stem active ingredient and sweroside were administered in amounts of 1.0g/kg, 1.5g/kg and 2.0g/kg, respectively, through the caudal vein of SD rats that had fasted for 4 hours (body weight: 120-170g, 5 male and female rats per administration, SLC, Japan). Rats were observed first for 30 minutes and then visually at 30 minute intervals. Mortality, general symptoms and weight changes were observed for 2 weeks after dosing. Physical dissection was performed to identify any organ abnormalities present.

The lethal dose of the honeysuckle stem active ingredient and the swertioside by oral administration exceeds 5.0g/kg (no dead rat is observed), and the lethal dose by intravenous injection exceeds 2.0g/kg (no dead rat is observed). When injected intravenously at 2.0g/kg, rats showed an increase in respiratory count and a decrease in activity, lasting about 10 minutes, with rapid recovery. No other symptoms were observed and no weight change occurred as a result of the administration. The results of the solid dissection showed no abnormalities, as in the case of the control group.

The local toxicity test at 50, 100 and 150 mg/dose, respectively, showed no difference from the group to which the physiological saline solution was administered. No toxicity such as tissue necrosis or inflammation was observed.

Preparation example 1: preparation of tablets

Preparing the active components of the honeysuckle stem or swertiamarin into tablets with the following compositions:

active component 160mg

Light anhydrous silicic acid 20mg

Corn starch 87mg

Crystalline cellulose 72mg

Glycolic acid (glyconate) starch sodium 60mg

Magnesium stearate 6mg

In total 672mg

Preparation example 2: preparation of syrup

Preparing the active components of the honeysuckle stem or swertia japonica Makino glycoside into syrup with the following components:

active ingredient 4,000mg

Methylparaben (5% ethanol solution) 60mg

Propyl p-hydroxybenzoate (5% ethanol solution) 40mg

Sodium benzoate (5% solution) 100mg

Banana powder (10% solution) 600mg

D-sorbitol 140,000mg

196mL of distilled water

Preparation example 3: preparation of injection

The honeysuckle stem active component or swertioside is prepared into an injection with the following composition:

injecting an ampoule: active ingredient 20mg

Mannitol 60mg

Corresponding solvent samples:

physiological saline solution for injection 2000mg

A total of 2080mg

Preparation example 4: preparation of injection

The honeysuckle stem active component or swertioside is prepared into an injection with the following composition:

injecting an ampoule: active ingredient 50mg

KH₂(PO₄) 8.5mg

Physiological saline solution for injection 3000mg

Total 3058.5mg

Preparation example 5: preparation of injection

The honeysuckle stem active component or swertioside is prepared into an injection with the following composition:

injecting an ampoule: active ingredient 100mg

Mannitol 300mg

KH₂(PO₄) 17mg

Physiological saline solution for injection 3000mg

A total of 3417mg

Preparation example 6: preparation of soft plaster

The active components of the honeysuckle stem or the swertioside are prepared into a soft plaster with the following composition:

active ingredient 5g

Liquid Paraffin 10g

9g of lead hard

Ethanol 8g

Sorbitan monooleate 2g

Polysorbate 4g

Propyl p-hydroxybenzoate 0.05g

0.1g of methylparaben

Concentrated glycerol 10g

Sufficient amount of pure water

As described above, the active ingredients obtained from honeysuckle stem according to the present invention have significantly increased solubility and blood stability compared to those commonly used. In addition, they were confirmed to have excellent analgesic and anti-inflammatory effects, safety and stability. In addition, the swertioside obtained from honeysuckle stem of the present invention has an extremely excellent medicinal effect and shows little toxicity, thus being particularly suitable for anti-inflammatory and analgesic drugs.

While the invention has been particularly described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes and substitutions may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (3)

1. The preparation method of the active components of the honeysuckle stem comprises the following steps:

(a) the method comprises the following steps: extracting honeysuckle stem with water under reflux, filtering, separating the layers by adding equal amount of saturated propanol or butanol solution to the filtrate, and concentrating the alcohol layer under reduced pressure to obtain primary active ingredient; and

(b) the method comprises the following steps: purifying the primary active ingredient with polyamide resin or polyvinylpyrrolidone resin to obtain a secondary active ingredient and further purifying with octadecylsilane resin to obtain a final active ingredient.

2. The preparation method of active constituents of honeysuckle stem according to claim 1, wherein the active constituents include swertioside and loganin as effective ingredients.

3. The preparation method of active constituents of honeysuckle stem according to claim 2, wherein the active constituents include swertiamarin 15.1-72.1 wt% and strychnine 13.9-41.4 wt%.