KR100868788B1 - PHARMACEUTICAL COMPOSITION FOR TREATING LONG QT SYNDROME COMPRISING AWF1k EXTRACT FROM ARTEMISIA IWAYOMOGI - Google Patents

Abstract

The present invention relates to a composition for treating QT interval prolonged syndrome, and more particularly, to a composition for treating QT interval prolonged syndrome, which contains the extract of A. erythema AWF1k as an active ingredient. AWF1k, the wormwood extract of the present invention, can be effectively used for the treatment and prevention of cardiomyocyte disease such as QT interval prolongation syndrome by activating ion channels in connection with diseases caused by inhibition of potassium channels.

Description

Composition for the treatment of PT interval prolongation containing Insam mugwort extract AHF1 {

1 is a graph showing the activity of the potassium ion channel recorded in one cardiomyocyte, Figure 1a is a graph showing the activity of the potassium ion channel for each voltage recorded after the AWF1k drug treatment, FIG. 1C is a graph showing the current-voltage relationship at the portion corresponding to the arrow of the small picture of FIGS. 1A and 1B by statistically processing the results of various cells.

Figure 2 is a graph showing the results of recording the activity voltage of one cardiomyocyte, Figure 2a is a graph showing that the action voltage period of the cardiomyocytes is reduced when AWF1k drug is applied, unlike the control group, Figure 2b is recorded in several cardiomyocytes A graph of the data statistically represented, where APD represents the active voltage period and APD ₅₀ represents the time it takes for 50% repolarization to complete after the active voltage begins to appear.

Figure 3 is a graph of the change in heart rate when the AWF1k drug is administered while perfusing the entire heart by the Langendorff method.

FIG. 4 is a graph showing the fraction of the Insam wormwood extract AWF1k obtained according to Example 1. FIG.

The present invention relates to a composition for treating long QT syndrome, and more particularly, to a composition for treating QT interval prolonged syndrome containing Artemisia iwayomogi extract AWF1k as an active ingredient.

Cardiovascular disease is a disease that is aggravated according to the manifestation and progression of the adult disease, a disease that fights the first and second place with cancer among the current causes of death. However, although various preventive measures and treatments for the development of the disease and the progression of the cardiovascular disease in Korea as well as in the world are being studied, the mortality rate or cure rate has not decreased. Therefore, the invention of a medicament for alleviating the symptoms of cardiovascular disease is still in high demand.

In order for the heart to move, a current must flow, which flows in a certain direction when the heart is normal and shows some pattern on the ECG. If there is an ischemic necrosis in the heart, or if there is an abnormality in the heart, such as an electrical wire, the pathological pattern appears on the ECG.

One of the most serious consequences of various heart diseases is the QT interval prolongation syndrome. QT interval prolongation syndrome is a congenital arrhythmia disorder mainly due to torsion de pointes, polymorphic ventricular tachycardia due to abnormal repolarization of the ventricles. Specifically, QT prolongation syndrome refers to an abnormality in the action voltage of ventricular cells, thereby extending the interval between ventricular depolarization and repolarization of the ventricles. The QT interval, which is the interval between the Q wave representing and the T wave representing repolarization of the ventricles, is extended. Extended QT intervals can cause severe ventricular tachycardia, which can be life-threatening, and the heart can suddenly stop and die suddenly, fainting, or fainting.

In most cases, the cause of this change is reported to be aberrant activity of potassium channels in various ion channels of ventricular cells that open and close according to the action voltage. Since the disease has no noticeable symptoms, the disease is often detected by an electrocardiogram test or a diagnosis test after a visit to the patient with fainting or fainting.

In addition, the QT interval prolonged syndrome is not significantly aware of the symptoms in the normal state, so it is not possible to give an accurate statistics on how many people have the disease, so there are various concerns about the severity of the disease.

With regard to the QT interval prolongation syndrome, the use of inhibitors for various potassium ion channels for the treatment is inevitable. In other words, there are no ion channels expressed only in the heart, and since the ion channels found to be one of the causes of the disease were not only present in the heart, potassium channels may cause symptoms, but the inhibitors cannot be administered. Therefore, there is an urgent need for the development of drugs that fundamentally treat QT interval prolongation syndrome and do not harm other organs.

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Injin mugwort is a perennial herb that belongs to the chrysanthemum family. This plant is an excellent source of green leaf protein, which is a very good food in terms of nutrition due to its high amount of essential fatty acids, ash and fiber (Ahn, BM, J. Korea Liver, 2000, 6 (4: 548 ~ 551); Hwang, et. al., J. Korean Soc.Nutr., 1998, 31 (4): 799-808; Nam, SM et al., J. Korean Soc.Food Sci.Nutr., 1998, 27 (2): 338-343 ). Previous studies reported on the efficacy of Injin mugwort include Kim, KS and Lee, MY, J. Korean Soc.Food Sci.Nutr., 1996, 25 (4): 581-587) and Lee, et al. GD et al., J. Korean Soc. Food Nutr., 1992, 21 (1): 17-22) reported that water extract or ethanol extract of mugwort had the effect of improving liver function in rats. Nam, SM et al., J. Korean Soc. Food Sci. Nutr., 1998, 27 (2): 338-343) and Lim, SS et al., J. Korean Soc. Food Nutr., 1997 , 30 (7): 797-802; Lim, SS and Lee, JH, J. Korean Nutr., 1997, 30 (3): 224-251) reported that hemoglobin and hepatic lipid levels decreased in hyperlipidemic rats. There is a bar. As described above, conventional studies have shown that the extract of Injin mugwort can be used for liver function improving agents or obesity treatment.

Injin mugwort is known to contribute to the overall immunity strengthening in the body, and its side effects are smaller than other drugs newly synthesized by chemical methods. However, there have been no reports that the extracts of Insam Worm can be used for diseases related to the heart.

Therefore, while the present inventors are studying various functions of the extract of Injin mugwort, which is known to have an effect on enhancing liver function and enhancing immunity, AWF1k, the water-soluble part of the extract of Injin mugwort, is effective in treating heart disease, especially QT interval prolongation syndrome. The present invention has been completed by discovering that it can act as a drug that does no harm to other organs.

It is an object of the present invention to provide a drug which can be used for the treatment of QT interval prolongation syndrome.

In order to achieve the above object, the present invention is a composition for treating QT interval prolongation syndrome containing the extract of Injin mugwort as an active ingredient, the extract of Injin mugwort is obtained by fumigation of dried Injin mugwort with high pressure steam to obtain an extract, and the extract is chromatographed. Fraction, which is prepared by concentrating, the prepared jinjin wormwood extract provides a composition for treating QT interval prolongation syndrome, characterized in that the fraction containing a carbohydrate absorbing a wavelength of 250 ~ 270 nm.
In addition, the present invention provides a composition for treating QT interval prolongation syndrome, characterized in that the extract of the ginseng mugwort is a mixture of small molecules of 1 kDa or less containing sugar residues and UV absorbing hydrophilic residues.

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The present invention provides a therapeutic composition that can be applied to the heart showing QT interval prolongation syndrome. More specifically, it suggests that the medicinal effect of AWF1k extract of Injin mugwort can be applied to QT interval prolongation syndrome.

On the other hand, the therapeutic composition comprising the jinjin Mugwort extract AWF1k of the present invention can be administered orally or parenterally during clinical administration and can be used in the form of a general pharmaceutical formulation.

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That is, the composition of the present invention may be administered in various formulations for oral and parenteral use in actual clinical administration, and when formulated, diluents such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. that are commonly used Or using excipients. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may comprise at least one excipient such as starch, calcium carbonate, sucrose in AWF1k extracts. ) Or lactose, gelatin and the like are mixed. In addition, various lubricants are used in addition to simple excipients. Liquid preparations for oral administration include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used. In addition, calcium or vitamin D ₃ may be added to enhance the efficacy of treating QT interval prolongation syndrome.

The dosage unit can be, for example, one, two, three or four times, or 1/2, 1/3 or 1/4 times the individual dosage. Individual dosages preferably contain an amount in which the effective drug is administered at one time, which usually corresponds to a full, 1/2, 1/3 or 1/4 times the daily dose.

The effective amount of the composition is 0.1-100 mg / kg, preferably 1-50 mg / kg, which may be administered 2-3 times a day. The composition as described above is not necessarily limited thereto, and may be changed depending on the condition of the patient and the onset of the disease.

As a result of oral administration and intraperitoneal administration of the pharmaceutical composition comprising the extract of Phosphorus mugwort of the present invention to mice, LD ₅₀ was found to be at least 10 g / kg or more, indicating a relatively safe substance.

AWF1k acquisition method according to the present invention is the same as the experimental procedure of the previous research paper (Koo, KA, et al., Arch. Pharm. Res., 1994, 17, 371374; Hwang, JS, et al., Arch. Pharm Res., 2003, 26, 294300; Ji, HJ, et al., Biotechnology Letters., 2005, 27, 253-257; Hwang, JS, et al, Biol. Pharm. Bull., 2005, 28, 921 -924). Specifically, the dried phosphorus mugwort is fumigated with high pressure steam to obtain an extract first, and the extract is distilled water as a developing solution, fractionated and concentrated by column chromatography using Sephadex G-50. Here, the fraction that absorbs a wavelength near 260 nm by analyzing the carbohydrates contained in each fraction by modified phenol sulfate assay and ultraviolet spectroscopy is named AWF1k. More precise analysis showed that most of these fractions are small compounds with sugar residues and hydrophilic residues that absorb ultraviolet light. Analysis by high performance size exclusion chromatography (Agilent) shows that AWF1k is a mixture of small molecules with molecular weight less than 1 kDa. In addition, AWF1k is a fraction concentrated to 50 mg / ml.

Tests were performed using the heart of New Zealand white rabbits using AWF1k, and the treatment of AWF1k increases the activity of potassium channels in cardiomyocytes (see FIG. 1) and shortens the interval of action voltage (see FIG. 2). ), Showing an effect of slightly increasing the heart rate (see FIG. 3).

Hereinafter, the present invention will be described in detail by the following examples.

However, the following examples are merely illustrative of one aspect of the present invention, and the content of the present invention is not limited by the following examples.

Example 1 Extraction of AWF1k

The extraction method of AWF1k was in accordance with the experimental procedure of the previous research paper (Koo, KA, et al., Arch. Pharm. Res., 1994, 17, 371374; Hwang, JS, et al., Arch. Pharm. Res., 2003, 26, 294300; Ji, HJ, et al., Biotechnology Letters., 2005, 27, 253-257; Hwang, JS, et al, Biol. Pharm. Bull., 2005, 28, 921-924).

Specifically, the dried phosphorus mugwort was fumigation with high pressure steam for 3 hours to obtain an extract. The extract was distilled water as a developing solution, and fractionated and concentrated by column chromatography using Sephadex G-50 (Koo, KA, et al., Arch. Pharm. Res., 1994, 17, 371374). The carbohydrates contained in each fraction were analyzed by modified phenol sulfate assay and ultraviolet spectroscopy. As a result, a fraction containing a large amount of carbohydrates and similar compounds, which is a fraction absorbing wavelengths near 260 nm, was named AWF1k (see FIG. 4). More precise analysis revealed that most of these fractions are small compounds (usually 2 kDa or less) that have sugar residues and hydrophilic residues that absorb ultraviolet light. As a result of high performance size exclusion chromatography (Agilent) using porous silicon microspheres (Zorbax, PSM60), it was found that AWF1k was a mixture of small molecules having a molecular weight of 1 kDa or less. AWF1k was obtained as a fraction concentrated at 50 mg / ml, and used in the experiment was diluted in each physiological saline solution.

Example 2 Experimental Animal

Each experiment was performed according to experimental animal care guidelines using the heart of a New Zealand white rabbit. The rabbits were raised to 2.0 kg with UV sterilized water and feed in an external and blocked environment where only purified air was allowed to enter before the experiment. According to the animal test instructions, anesthetics (Entoval 50 mg / kg) and heparin (100 U / kg) were injected through the ear vein, anesthetized, and the heart was taken out and used for the experiment.

Example 3 Heart Test with Extract of AWF1k

The extract of AWF1k was tested on the heart obtained in Example 2. AWF1k was used as a 1000-fold dilution (50 ㎍ / ㎖) solution, the control was confirmed by the addition of the same amount of distilled water.

Experiments on the activity of the potassium channel (FIG. 1) and the action voltage (FIG. 2) were performed as cardiomyocytes digested with collagenase and separated into single cells using the Langendorf method. In addition, using the Langendorf method, the heart was measured after allowing the saline to enter and exit the heart similar to the physiological state (FIG. 3).

Specifically, Figure 1 is a graph showing the activity of potassium ion channels recorded in one cardiomyocyte. At this time, Figure 1a is a graph showing the activity of the potassium ion channel for each voltage recorded after the drug is added, respectively, after treatment with the AWF1k extract drug, Figure 1c is a small arrow of Figures 1a and 1b The current-voltage relationship at that point is statistically represented by integrating the results from several cells. The change in the potassium channel in the cardiomyocytes is a measure of the current, which is the flow of ions moving in and out of the cell membrane using an experimental technique called voltage-fixing (FIG. 1). As a result, treatment with AWF1k increased the activity of potassium channels in cardiomyocytes compared to controls (FIG. 1).

Figure 2 is a graph showing the results of recording the activity voltage of one cardiomyocyte. Figure 2a shows that the action voltage period of the cardiomyocytes is reduced when AWF1k drug is added unlike the control group, Figure 2b is a statistical process of the data recorded in several cardiomyocytes. At this time, APD represents the active voltage period, APD ₅₀ represents the time taken to complete 50% repolarization after the active voltage starts to appear. The activation voltage of the cardiomyocytes was measured by the voltage difference between the cell and the ophthalmology on the basis of the cell using an experimental technique called voltage-fixing method (FIG. 2). As a result, treatment with AWF1k resulted in shorter intervals of action voltage compared to controls (FIG. 2).

Figure 3 shows the change in heart rate when administration of the AWF1k drug while perfused throughout the heart by the Langendorf method, AWF1k treatment showed an effect of slightly increasing the heart rate compared to the control (Fig. 3).

As such, it was confirmed that the current of ions through the potassium channel increases in size with the addition of AWF1k in a situation similar to the physiological environment inside and outside the cells. This increase in potassium current affects the action voltage and results in a change in repolarization, in particular referred to as phase 3. That is, as shown in FIG. 2, it is determined that repolarization occurs quickly to reduce the interval of the active voltage, thereby reducing the inconsistency caused by the interval of the active voltage.

Example 4 Acute Toxicity Test with AWF1k Extract

Phosphorus mugwort used in the present invention was widely used for food, so it was judged that there was no problem in stability, but the oral administration and the intraperitoneal administration were carried out to determine the toxicity.

Acute toxicity experiments were performed using 6-week-old SPF SD rats. Two animals per group were suspended orally at a dose of 10 g / kg in AWF1k of the present invention, each suspended in 0.5% methylcellulose solution. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed. Hematological and hematological examinations were performed. Necropsy was performed to visually check for abdominal and thoracic organ abnormalities.

As a result, no significant clinical symptoms or dead animals were noted in all animals treated with the test substance, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings. The results showed that AWF1k did not show toxic changes in all rats up to 10 g / kg and LD ₅₀ was determined to be a safe substance of 10 g / kg or more.

As described above, the AWF1k, the extract of the present invention, AWF1k, can be effectively used for the treatment and prevention of cardiomyocyte disease by activating ion channels for diseases caused by inhibition of potassium channels, such as QT interval prolongation syndrome.

Claims (5)

As a composition for the treatment of QT interval prolongation syndrome, which contains the extract of Insam wormwood as an active ingredient, The extract of Injin mugwort is prepared by fumigation of dried Injin mugwort with high pressure steam to obtain an extract, fractionating and extracting the extract by chromatography, The prepared jinjin mugwort extract is a composition for treating QT interval extension syndrome, characterized in that the fraction containing a carbohydrate absorbing a wavelength of 250 ~ 270 nm. delete delete delete The composition of claim 1, wherein the extract of the ginseng wormwood is a mixture of small molecules of 1 kDa or less containing sugar residues and UV absorbing hydrophilic residues.

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