KR100259175B1 - The use of l-muscone - Google Patents

Abstract

PURPOSE: Medical use of l-muscone is provided to be variously used as a heartometer/circulatory system agonist, a brain disease treatment, and a central nervous system agonist instead of musk. CONSTITUTION: The l-muscone is able to be used instead of musk in the manufacture of various kinds of herb medicine. The one day dosage of the l-muscone is 50 micrograms to 10,000 micrograms, and preferably 50 micrograms to 1000 micrograms. The l-muscone is medicated via an oral administration manner, an intravenous injection manner, and the other possible clinical channel. The l-muscone has many functions such as a blood vessel relaxation function, a blood pressure drop function, a palpitation suppression function, and shows a superior treatment effect to transient cerebral ischemia attack and stroke. Also, the l-muscone has a panting effect, an ultromotivity reduction effect, and a stress suppression effect.

Description

Medicinal Uses of L-Muscon {The use of l-muscone}

The present invention relates to the medical use of l-muscone having a prophylactic and therapeutic effect on the cardiovascular system, the central nervous system and brain diseases.

Musk is traditionally used as a fragrance regulation in oriental medicine with cow sulfur, cephalophyllus, spearmint, benzoin, and mussel oil (synthetic fragrance). As an important expensive herbal medicine that has been widely used in the back, musk's medicinal effects and pharmacological effects are reported to have cardiovascular, central nervous system, antibacterial, anti-inflammatory, androgenic hormones, antithrombotic and detoxifying effects. It became.

The main pharmacological action of musk is expressed by which components of musk constituents. Among them, muscone, the main ingredient of musk, has been studied for its efficacy and pharmacology. Only sedation results were reported. In the case of sedation, the muscone showed significant sedation, especially in the state of exercise excitement. There have been reports of apomorphine-like action (Kimura et al., Journal of Medical and Pharmaceutical Society for Wakan-Yaku 3,324-325, 1986), which shows excitability. Increased vascular blood flow in Langendorf specimens and dose-dependent heart rate in atrial specimens Increases were reported (Toyoguchi et al., Applied Pharmacology and Therapeutics, 33 (4) 701-706, 1987).

In addition, transient anesthesia and dose-dependent bradycardia were observed in anesthetized rats, and the cardiac activity was apparent due to the decrease in the number of beats of the atrial myocardium and the increase in contraction tension. There have been reports of a decrease and an increase in respiratory amplitude (Matsubara, 1990).

For detoxification and drug metabolism, hepatic metabolizing enzyme cytochrome P-450 is induced (Peng et al., 1986), and this form of induction is Similar results were obtained after treatment with phenobarbital (Tanaka et al., 1987; Tanaka et al., Biochem. Pharmacol. 41 (3), 472-473, 1991).

Muscone used in the above-described experiments (physicochemical) d-, l- or d, l-form (form), such as no specific reference, so the pharmacological test results could not be accurately determined.

Therefore, in order to verify the efficacy and effects of muscone, we have studied the pharmacological action for several years with L-muscone, the main ingredient of natural musk, and the cardiovascular system, which is the main action of musk. It is concluded that the present invention is effective in cardiovascular system, central nervous system, brain, respiratory system, blood vessel relaxation, blood pressure lowering and cardiac contractility, heart function (contraction and contraction frequency to heart), central nervous system, brain disease and respiratory system. This study was completed so that it could be used as a raw material of medicines with the effect of promoting diseases and hops.

L-muscone, which has such musk yang efficacy and effect, has not been put to practical use because it is almost impossible to mass-produce by simple synthetic method, but at the end of 10 years of research in this company, dl-muscone has high yield and high purity. And successful methods of mass production at low cost (Patent Application Nos .: 22981 (94.9.12), 4715 (96.2.27) and 4716 (96.2.27)). By allowing mass production of l-muscone [Patent Application No. 43,44,45 (97.1.4)], various stress and natural environment caused by the complicated daily life of society Other adult diseases (e.g. hypertension, stroke etc., cardiac disorders, cardiac palpitations, respiratory distress, neurological disorders such as acute ailment, etc.) such as mental illness and autonomic anxiety. As a raw material for new ingredients used to improve symptoms or for treatment It is a feature of the present invention to develop a new use of l-muscone for use.

1 is the effect on the mean blood pressure and heart rate when administered intravenously in cats anesthetized L-muscone (l-muscone).

Figure 2 is the effect of blood pressure upon intravenous administration to cats anesthetized L-muscone (l-muscone).

FIG. 3 shows the effect of l-muscone on cardiac contractility in the extracted right atrium of rabbits.

Figure 4 is an effect on the heart rate of l-muscone in the extraction right atrium of rabbits.

5 is a glass variation of [ ³ H] 5-HT caused by the hypoxic state.

6 is the effect of l-muscone on the [ ³ H] 5-HT glass fluctuations in the normal and hypoxic state.

Figure 7 is a comparison of the increase rate of [ ³ H] 5-HT glass by the administration of l-muscone in normal and hypoxic state.

Figure 8 is the effect of l-muscone on blood pressure in congenital hypertensive rats.

9 is the effect of l-muscone on heart rate in congenital hypertensive rats.

10 is the effect of inhibiting palpitations of l-muscone (l-muscone).

11 is the effect of l-muscone on heart rate in congenital hypertensive rats.

The present invention relates to the use as a cardiovascular system having l-muscone as a main component, and has a blood vessel relaxation, blood pressure lowering and palpitation-inhibiting effect, and also contains L-muscone as a main component. The present invention relates to a brain disease agent having a therapeutic effect on cerebral ischemia and stroke, and another use of the present invention is a respiratory accelerator and a sedative agent having a therapeutic effect on respiratory distress, which contains l-muscone as a main ingredient. It includes the use as a central nervous system solvent having a spontaneous motor activity reduction and stress inhibitory effect.

L-muscone of the present invention has the advantage that can be used instead of musk in the manufacture of several traditional herbal preparations such as bovine sulfur cheongsimwon, Ki Eunghwan, centripet and musk lipogen, the daily dosage is 50㎍ ~ 10,000㎍ to use But preferably 50-1000 μg. Routes of administration include all possible routes of administration in oral, intravenous and other clinical routes.

Hereinafter, experimental examples and preparation examples of the effect of L-muscone will be described, and the scope of protection of the present invention is not limited thereto.

Experimental Example 1

Effect on the cardiovascular system (1)

1. Experimental Materials and Methods

1) experimental animals

Healthy cats and rabbits were purchased from Cheil Corp. and used in the experiments for one week in the GLP room.

2) Blood pressure and heart rate measurement

Cats were intraperitoneally injected with 35 mg / kg of secobarbital sodium and anesthesia was used to secure airways using tracheotomy tubes to prevent death from secretions. Physiological saline containing heparin (200 unit / ml) to expose one femoral artery, insert an arterial cannula, connect it to a pressure transducer, and prevent blood from clotting between the pressure transducer and the cannula After filling the connection site, the mean blood pressure and heart rate from the femoral artery were measured. The cannula with 3-way stopcock attached to the same femoral vein was used to inject the drug, and all the drugs were injected slowly and the continuous blood pressure and heart rate were measured. All the drugs were injected with a little saline for injection so that they could not remain in the cannula after the injection, and care was taken not to raise blood pressure due to sudden external injection.

3) Effect of l-muscone on blood pressure and heart rate

L-muscone was suspended in 10% tween 80 and used for the experiment. L-muscone suspensions were slowly injected into the cannula of the femoral vein at concentrations of 50 and 100 mg / kg and the changes in blood pressure and heart rate were observed.

4) Influence of l-muscone on the harvested atrium of rabbits

The rabbit's heart is removed and blood is removed from a solution of Klebs-Henseleite saturated with 95% O ₂ /5% CO ₂ .The right atrium is quickly separated and the ends are threaded and one end The other one was fixed to the isometric transducer. Basal resting tension was adjusted to 500 mg tension and waited for about 1 hour until the tissue stabilized and used in the experiment. At this time, the temperature of the nutrient solution was maintained at 37 ℃ similar to the temperature in vivo. L-muscone was dissolved in 75% DMSO and used in all experiments. The final concentration of DMSO in the organ bath was adjusted to 0.2% or less. After l-muscone is administered at a concentration of 100 µg / ml, the drug is allowed to be absorbed for 15 minutes, and isoproterenol is cumulative at a concentration of 10 ^-9 to 10 ^-7 M for 3 minutes. Administration at intervals of less than one hour measured the contractile force and contraction frequency of the right atrium. The control group was administered through the same experimental procedure after administration of the same concentration of DMSO. The experimental results for the cardiac contractile force were expressed in relative proportions with the maximum contraction force being 100%, and the experimental results for the contraction frequency were expressed in the number of contractions per minute.

2. Experimental results

1) Effect of L-muscone on Cardiovascular System and Dose Response

Intravenous injections of L-muscone 50 mg / kg and 100 mg / kg showed a mean drop in mean blood pressure of 29 mmHg and 48 mmHg, respectively. On the other hand, in the heart rate, as a slight dose-dependent reduction effect, it was confirmed that the heart rate decreased by about 14 times and 35 times per minute at 50 mg / kg and 100 mg / kg, respectively (FIGS. 1 and 2).

2) Effect of l-muscone on the extraction right atrium of rabbits

When the extracted right atrium of the rabbit isopropyl furo terephthalate play (isoproterenol) hayeoteul Continue cumulative administered at 10 ^-9 M were able to confirm the positive inotropic effect and a positive chronotropic effect, it showed a maximum shrinkage force when hayeoteul administration of 10 ^-7 M, At higher concentrations, arrhythmia occurred, so the experiment was conducted with the highest isoproterenol dosage of 10 ^-7 M. Treatment with DMSO (less than 0.2%) and l-muscone (100 µg / ml) in the normal harvested atrial appendage had no effect on contractile force and contraction frequency. However, pretreatment with l-muscone significantly reduced the positive inotropic and positive chronotropic effects induced by isoproterenol (FIG. 3, 4).

3. Consideration

In the above results, L-muscone, the main ingredient of musk, decreases blood pressure and heart rate in the cardiovascular system and does not affect the heart in normal condition, but isoproterenol increases cardiac contractility and contraction. The effect of reducing the maximum frequency was found to be effective in calming the excited state such as palpitation.

Experimental Example 2

Impact on cerebral ischemia

1. Experimental Materials and Methods

1) Experiment animal

Sprague-Dawley male rats (200-250 g) were acclimated for one week in the GLP room and used for the experiment.

2) Reagent

5-Hydroxy [G- ³ H] tryptamine creatinine sulfate ([ ³ H] 5-HT) is from Amersham, nialamide is from Sigma, zimelidine Silver was purchased from Research Biochemical and used for the experiment.

3) Production of hippocampus slices

After the rats were sacrificed by the guillotine, the brains were quickly removed, the cerebral hemispheres were cut on the ice plate, and the cerebral cortex was removed to obtain the whole hippocampus. Hippocampal tissues were cut to a thickness of 400 μm in the transverse direction using a Mcllwain mechanical tissue chopper (The Mickle Laboratory Engineering Co., Gomshall, Surrey, UK) to obtain hippocampal sections and saturated with 95% O ₂ and 5% CO ₂ . Tissue debris was removed by suspending in cold nutrient solution [NaCl 124mM, KCl 4mM, CaCl ₂ 2mM, KH ₂ PO ₄ 1.25mM, MgSO ₄ 1.0mM, NaHCO ₃ 25mM, glucose 10mM, Nialamide 12.5μM].

4) Adsorption of [ ³ H] 5-HT and Induced Hypoxia

Hippocampal sections were stabilized for 30 minutes in a nutrient solution at 37 ° C. and adsorbed to the tissue for 20 minutes in a nutrient solution containing 0.1 μM of [ ³ H] 5-HT. The hippocampal tissues were washed three times with the same nutrient solution to remove unadsorbed [ ³ H] 5-HT, and then 3 ml of fresh nutrient solution was added thereto and incubated for 10 minutes at 10 minute intervals. At this time, 10 μM of zimelidine was added to the nutrient solution used to prevent re-adsorption of free [ ³ H] 5-HT. Hypoxia was induced by exchanging nutrient solution saturated with 95% N ₂ and 5% CO ₂ in the 60 min and 70 min fractions, and test specimens were also treated at the same time.

5) Determination of liberated [ ³ H] 5-HT

Tissue dissolving agent (Soluen 350, packard Instrument Company, Inc., Downers Grove, IL, USA) was added to the cultured hippocampal tissues, dissolved at 47 ° C for 1 hour, and neutralized by adding 50 µl of glacial acetic acid. Take 1 ml of the culture solution and 0.1 ml of tissue solution, mix well with 9 ml of radioactivity measurement solution (READY SAFE, Beckman Instrument Inc., Fulletron, CA, USA), and then use Liguid Scintillation Counter (Beckman Instrument Inc., Fulletron, CA, USA). The amount of free [ ³ H] 5-HT is expressed as a percentage of the amount adsorbed on hippocampal tissue.

6) Statistics processing

Each result was expressed as mean ± standard error. Statistical significance was verified by analysis of variance (ANOVA) and the difference between groups was compared using Student New-Man Keul test.

2. Experimental Results

1) [ ³ H] 5-HT glass fluctuation caused by hypoxia

The spontaneous release of [ ³ H] 5-HT in the hippocampal section continued, with a rapid decrease in glass initially and a constant glass pattern after 40 minutes (Figure 5). Therefore, in this experiment, the free amount of [ ³ H] 5-HT was expressed as a relative value based on the free amount at 50 minutes, induced hypoxia at 50 and 60 minutes after incubation, and the test specimens were treated. Spontaneous release of [ ³ H] 5-HT was markedly reduced when hypoxia was induced, and returned to normal when oxygen was resupplied.

2) Effect of l-muscone on [ ³ H] 5-HT Glass Fluctuation

When L-muscone, the main component of musk, was administered at a concentration of 10 ^-5 M in normal oxygen, the release of [ ³ H] 5-HT increased significantly compared to the control, and also in hypoxic condition. Significantly increased free to recover similar to normal oxygen control (FIG. 6). Comparing the increase rate of [ ³ H] 5-HT glass by the administration of l-muscone in the normal and hypoxic state showed more free increase in the hypoxic state (FIG. 7).

3. Consideration

In the above experimental results, it was confirmed that the effect of restoring 5-HT free reduction caused by hypoxia to the normal state when L-muscone, the main component of musk, was administered. In addition, when comparing the increase rate of 5-HT glass in the normal oxygen state and low oxygen state, it was confirmed that L-muscone increased more in the low oxygen state. Therefore, L-muscone, the main component of musk, has an effect of normally restoring the free decrease of 5-HT induced by cerebral ischemia, and L-muscon (l-muscone) having the 5-HT free increase action (l-muscone). -muscone may reduce the nerve damage caused by cerebral ischemia.

Experimental Example 3

Impact on stroke

In histopathologic examination, cell damage due to cerebral ischemia and reperfusion was inhibited in the L-muscone group, and ATP content inhibited ATP reduction due to cerebral ischemia and reperfusion in the L-muscone group. ,

Lactate content showed a significant lactic acid inhibitory effect compared to the control group, Lipid peroxidation showed a significant decrease compared to the control group was confirmed to be effective in stroke (see Table 1 and 2).

TABLE 1

TABLE 2

Experimental Example 4

Impact on the cardiovascular system (2)

1) Effect on hypertension

As a result of response test for blood pressure of congenital hypertensive rat (SHR), L-muscone showed significant decrease in blood pressure and heart rate in comparison with control group (Fig. 8 and Fig. 8). 9)

2) Effect on palpitations

As a result of response test against palpitation in normal rats, L-muscone showed a good palpulation inhibitory effect, and showed a significant inhibitory effect on the heart rate change rate compared to the control group (FIGS. 10 and 11). Reference)

Experimental Example 5

Experiment on the action on the respiratory system

1. effects on dyspnea;

For urethane suppressed breathing, the l-muscone group showed an increase in respiratory rate and respiratory depth, and the respiratory excitability was recognized (see Table 3).

TABLE 3

Experimental Example 6

Experiments on the central nervous system

1. effect on soothing action;

Rotarod test showed significant sedation in the L-muscone group 0.45 and 4.5 mg / kg compared to the normal group (see Table 4).

TABLE 4

Significantly different from the normol group (*; p <0.05, **; p <0.01 in x ² -test)

2. effect on spontaneous motor performance;

In the L-muscone group, the 0.15, 1.5 mg / kg administration group showed a significant decrease in spontaneous motor performance compared to the normal group (see Table 5).

TABLE 5

Significantly different from the normol group (*; p <0.05, **; p <0.01 in t-test)

3. action on antistress;

① The weight of the adrenal gland that was significantly enlarged after stress was significantly inhibited in the l-muscone 1.5 and 4.5 mg / kg groups compared with the control group.

② Significant decrease in the amount of ascorbic acid in the adrenal glands due to stress was significantly suppressed in the L-muscone 1.5 mg / kg administration group (see Table 6).

TABLE 6

Values are means ± S.E.M for 7 rats per group

Significantly different from the control group (*; p <0.05, **; p <0.01 in t-test)

Significantly different from the normal group (#; p 〈0.05, ##; p 〈0.01 in t-test)

Experimental Example 7

L-muscone Acute Toxicity Test

1. Sample: l-muscone

2. Experimental Animal

Rats: SPF rats of the 5 week old SD strain

Male: 127.2 ~ 144.7g

Female: 112.5 ~ 138.2g

The experimental animals were freely fed solid feed and water under conditions of room temperature 23 ± 3 ° C., humidity 50 ± 10%, illumination time 12 hours (7 am to 7 pm and illuminance 150 to 300 Lux).

3. Administration method and observation

Five rats of one group were used, and the animals were fasted overnight before administration, and the test substance was orally administered using a syringe equipped with an oral dose of Sonde. Changes in general symptoms were observed at least once a day from the next day to 14 days from 1 hour to 6 hours after administration. LD ₅₀ was not observed because no dead animals were observed.

4. Experimental Results: The experimental results are shown in Table 7 below.

TABLE 7

From the experimental results, it can be seen that the acute toxicity of the l-muscone of the present invention can be ignored.

(Example 1-4)

1) Xinwu Hwang Cheongsimwon in 1 pill, 282 mg, licorice 202 mg, ginseng 97 mg, turmeric 100 mg, singok 100 mg, soybean yellow soybean 70 mg, cinnamon 70 mg, glue 70 mg, peony 60 mg, Mcmundong 60 mg, golden 60 mg , Angelica 60mg, Windproof 60mg, Baekchul 60mg, Siho 50mg, Gilgyeong 50mg, Pedestrian 50mg, Bokryeong 50mg, Cheongung 50mg, Sulfur 14mg, Nutrition angle 35mg, Longpan 41mg, Whiteworm 30mg, Health 30 mg, l-muscone (75 μg) was prepared according to the preparation of pills.

2) Sinwonbang Hwang Cheongsimwon in one round of medicinal herbs 263 mg, licorice 188 mg, ginseng 94 mg, turmeric 94 mg, singok 94 mg, soybean yellow soybean 66 mg, cinnamon 66 mg, glue 66 mg, peony 56 mg, Mcmundong 56 mg, golden 56 Mg, Angelica 56 mg, Windproof 56 mg, Baekchul 56 mg, Shiho 47 mg, Gil Gyeong 47 mg, Pedestrian 47 mg, Bokryeong 47 mg, Cheongung 47 mg, Sulfur 45 mg, Nutrition angle 38 mg, Long brain 38 mg, White rot 28 mg, Health 28 mg, l-muscone 570 μg was prepared according to the preparation method of the pills.

3) Xinwu Hwang Cheong-Shim Cheong-Sen Solution 50ml, 282mg, Licorice 202mg, Ginseng 97mg, Sulfur 100mg, Singok 100mg, 70g Soybean Curd 70mg, Cinnamon 70mg, Glue 70mg, Peony 60mg, Mcmundong 60mg, Golden 60 Mg, Angelica 60 mg, Windproof 60 mg, Baekchul 60 mg, Shiho 50 mg, Gil Gyeong 50 mg, Pedestrian 50 mg, Bokryeong 50 mg, Cheongung 50 mg, Cow sulfur 14 mg, Nutrition angle 35 mg, Long brain 41 mg, White worm 30 mg, Health 30 mg, l-muscone (75 μg) is prepared according to the preparation method of the liquid formulation.

4) Sinjang 263 mg, licorice 188 mg, ginseng 94 mg, turmeric 94 mg, singok 94 mg, soybean yellow soybean 66 mg, cinnamon 66 mg, glue 66 mg, peony 56 mg, macmundong 56 mg, gold 56mg, Angelica 56mg, Windproof 56mg, Baekchul 56mg, Shiho 47mg, Gilgyeong 47mg, Pedestrian 47mg, Bokryeong 47mg, Cheongung 47mg, Sulfur 45mg, Nutrition angle 38mg, Longpan 38mg, Whiteworm 28mg , 28 mg of health, 570 μl of L-muscone are prepared according to the preparation method of the liquid formulation.

The present invention is effective in treating cardiovascular disease (vascular relaxation, blood pressure lowering and antihypertensive effect), brain disease (cerebral ischemia, stroke), dyspnea and central nervous system (calm, spontaneous motor loss, stress suppression effect) disease It relates to the use of l-muscone, and the daily dose is 50 to 1000 µg.

Claims (3)

A respiratory accelerator that has a therapeutic effect on respiratory distress, which contains L-muscone as a main ingredient. Central nervous system solvent with l-muscone (l-muscone) as the main ingredient, has a sedation, spontaneous motor activity and stress inhibitory effect. The respiratory accelerator or central nervous system solvent according to any one of claims 1 to 3, wherein the daily dose is 50 to 1000 µg.