KR101828102B1 - Composition for treating vomiting or diarrhea comprising Machili Thunbergi Cortex extract - Google Patents

Abstract

The present invention is directed to a composition for preventing, ameliorating and / or treating vomiting or diarrhea, including tobacco extract. The compositions of the present invention exhibit a preventive, ameliorative, and / or therapeutic effect of vomiting or diarrhea due to vomiting or diarrhea suppressed by a 5-HT ₃ receptor antagonist, especially a chemotherapeutic agent, an irradiation treatment, or anesthetic administration , Which selectively antagonize the 5-HT ₃ receptor and do not act on dopamine receptors and do not cause side effects.

Description

TECHNICAL FIELD The present invention relates to a composition for treating vomiting or diarrhea,

The present invention is directed to a composition for preventing, ameliorating and / or treating vomiting or diarrhea, including tobacco extract.

According to statistics by the National Statistical Office (NSO), Korea's mortality rate was 142.4 per 100,000 people in 2011, compared with 144.4 per 100,000 in 2010. In addition, the number of cancer incidence has increased annually, and in 2010, it increased to 202.05 per 100,000 people, up 3.7% from the previous year (National Cancer Information Center).

Cancer mortality is declining due to the development of chemotherapeutic drugs such as chemotherapeutics, radiation therapy, etc. However, side effects (nausea, vomiting, and diarrhea) caused by chemotherapy still cause great pain for patients have. All vomiting is controlled by vomiting in the retina. The chemoreceptor trigger zone (CTZ) and isolated nucleus (NTS) located in the lower zone of the fourth ventricle are the most important relay centers of afferent stimuli in the periphery. In addition, CTZ is not protected by the cerebrovascular barrier, and it secretes neurotransmitters by directly sensing toxins in blood and cerebrospinal fluid (Mahesh, Perumal et al., 2005).

Vomiting and diarrhea are very diverse, and research has shown that vomiting or diarrhea caused by cytotoxic drugs such as chemotherapy is associated with increased 5-HT (5-hydroxytryptamine, or serotonin) levels in intestinal, blood, and brain stem (Minami, Endo et al., 2003). The cytotoxic chemotherapeutic agent increases the 5-HT release of enterochromaffin cells in the intestinal mucosa and the increased 5-HT binds the 5-HT ₃ receptor of the afferent vagal nerve endings to stimulate the brain stem Or vomiting to cause vomiting reflections (Matsuki 1996).

Vomiting and diarrhea occur in the case of radiation treatment and after administration of anesthetic. Although the cause of this is unclear, it is thought that 5-HT induction current is generated by 5-HT ₃ receptor due to treatment with 5-HT ₃ receptor selective antagonist.

Such vomiting or diarrhea can not be treated with antibiotics used to treat vomiting or diarrhea due to bacterial infection or the like and can not be treated with dimenhydrinate used for the treatment of vomiting caused by vestibular stimulation such as motion sickness . Therefore, the 5-HT ₃ receptor antagonist is used to treat the dopamine receptor due to the structural similarity between serotonin and dopamine drugs, and various side effects occur.

Ondansetron is a 5-HT ₃ receptor selective antagonist commonly used in the prevention or treatment of diarrhea caused by vomiting or diarrhea, irritable bowel syndrome induced by chemotherapeutic agents, radiation therapy, and anesthetic administration in clinical practice. Ondansetron induces vomiting by inhibiting the 5-HT ₃ receptor in the visceral centromere (Andrews, Davis et al., 1990). However, some of these drugs are accompanied by side effects, and fewer side effects are given to the patients because of the economic burden on the patient. Since the market of antiseptic drugs and cancer chemotherapy market are interdependent, it is now a reality that antiseptic agents which have few side effects are not only expensive but also have a limited amount of insurance. Therefore, the research and development of an anti-aging agent which is low in side effects and economical is attracting attention.

The inventors of the SAT magnolia extract the 5-HT ₃ receptor underwater, as a result of research on herbal medicines, known as non-toxic in traditional medicine, and who wish to develop herbal medicines that can be, I sat port to replace the ondansetron It was confirmed that the selective antagonistic effect was excellent, and the present invention was completed.

The present invention is intended to provide a composition for treating vomiting or diarrhea.

As described above in the background art of the present invention, since the action mechanism differs depending on the type of vomiting or diarrhea, development of a therapeutic agent acting on a specific mechanism has been required. The inventors of the present invention discovered that 5-HT induction current suppression effect of 5-HT induced current suppression effect was particularly excellent when the vagus nerve cells were isolated and treated with various herbal medicine extracts, Or diarrhea is effective for the first time.

The present invention provides a pharmaceutical composition for prevention or treatment of vomiting or diarrhea, which comprises a tofu extract as an active ingredient. The tobacco extract has a selective antagonistic effect on the 5-HT ₃ receptor present in the vomiting center of the gastrointestinal tract or soft drink. Preferably, the vomiting or diarrhea is caused by an increase in 5-HT concentration, Radiation therapy, or vomiting or diarrhea due to awakening after administration of anesthetic.

When 5-HT binds to the 5-HT ₃ receptor at the vagal nerve endings, signals are transmitted to the brainstem or vomitus center, causing vomiting or diarrhea. The composition of the present invention inhibits vomiting or diarrhea by binding to 5-HT ₃ receptors and blocking 5-HT binding. However, the composition of the present invention has an advantage that it does not bind to a dopamine receptor similar to the 5-HT ₃ receptor and does not cause various side effects due to the generation of dopamine induced current.

Machili Thunbergi Cortex is a bark of Machilus thunbergii Sieb. Et ZUC c. And Machilus thunbergii var. Obovata Nakai (Lee., Kim et al. 2008). Tofu is not used as Chinese medicine in Chinese and Japanese medicine, but it is used in Korea as a substitute for Chinese medicine. (Li, Ju et al., 2003), inhibition of gastrointestinal tract function, suppression and relaxation of intestinal motility, and gastric juice And suppression of gastric acid secretion, increased bile secretion, inhibition of shay and drug-induced gastric ulcer, and antibacterial activity against intestinal bacteria (Lee and Chung, 1994). In addition, it has been reported that the extract of Tobacco extract inhibits the phase contraction of the mouse small intestine and differs from that of the extract (Lee, Park, et al., 2011).

The tofu extract of the present invention is extracted with an extraction solvent selected from water, C1 to C4 alcohols or a mixture thereof, and may be extracted using other conventional solvents in the art. The extraction method may be a method commonly used in the art, for example, ethanol extraction, water extraction, or both heat extraction and cold extraction. Water used as the solvent is not limited to the kind of water, but it may be preferable to extract distilled water as a solvent. In one example of the present invention, heat extraction by water is performed as follows. The resulting mixture is pulverized by hot air drying at 30 to 70 캜, preferably 40 to 60 캜, and then put into 0.1 to 10 L of distilled water, preferably 3 to 5 L of distilled water per 100 g of the powder, At 80 to 120 ° C, preferably 90 to 110 ° C for 70 to 120 minutes, preferably 80 to 100 minutes. Further, after the extraction, it was allowed to cool at room temperature, 1 filter paper. The obtained residue is mixed with 0.1 L to 3 L of distilled water, preferably 0.5 to 2 L of distilled water, and then filtered at 80 to 120 ° C, preferably 90 to 110 ° C for 70 to 120 minutes , Preferably 80 to 100 minutes, followed by cooling at room temperature. 1 filtration using a filter paper. In the present invention, cold extraction using water can be used. As an example of the present invention, the tofu cake is pulverized by hot air drying at 30 to 70 DEG C, preferably 40 to 60 DEG C, L to 10 L of distilled water, preferably 3 L to 5 L of distilled water, at 0 to 10 캜, preferably 2 to 6 캜, more preferably 4 캜 for 12 to 48 hours, preferably 18 to 30 And further preferably 24 hours. After extracting, it is allowed to cool at room temperature and then filtered under reduced pressure using Whatman Grade No.1 filter paper. The obtained residue is dissolved in 0.1 L to 3 L of distilled water, 2 L of distilled water and then extracted at 0 to 10 ° C., preferably 2 to 6 ° C., more preferably 4 ° C. for 12 to 48 hours, preferably 18 to 30 hours, more preferably 24 hours. Grade No.1 F Whether it is possible to extract in a way that filtered using reduced pressure. The extraction method of the present invention is not limited to the above example.

The pharmaceutical composition of the present invention may be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.

The composition according to the present invention may be formulated into pharmaceutical compositions containing at least one pharmaceutically acceptable carrier in addition to the above-described effective ingredients for administration.

Formulations of the pharmaceutical compositions of the present invention may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, .

For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, natural sugars such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum and the like.

Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The composition or the active ingredient of the present invention can be administered orally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, nasally, by inhalation, topically, rectally, , And can be administered intravenously, preferably.

In the composition of the present invention, the dose of the active ingredient can be adjusted within the range of 0.5 mg / kg to 50 mg / kg on the basis of 60 kg body weight. However, the optimal dose to be administered can be easily determined by those skilled in the art, and can be easily determined by a person skilled in the art and depends on various factors such as the type of disease, the severity of the disease, the content of the active ingredient and other ingredients contained in the composition, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like.

The present invention also relates to a method of treating diarrhea or vomiting comprising administering to a mammal a therapeutically effective amount of a tobacco extract, preferably diarrhea or vomiting due to an increased 5-HT concentration, more preferably a chemotherapeutic agent, Or treatment of vomiting or diarrhea caused by arousal after anesthesia treatment or irradiation therapy.

As used herein, "therapeutically effective amount" means the amount of active ingredient or pharmaceutical composition that elicits a biological or medical response in an animal or human considered by a researcher, veterinarian, physician or other clinician, And an amount that induces symptomatic relief of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect.

The present invention also provides a food composition for preventing or ameliorating diarrhea or vomiting, preferably vomiting or diarrhea due to an increase in 5-HT concentration, which contains as an active ingredient a tofu extract, Vomiting or diarrhea is vomiting or diarrhea due to awakening following administration of a chemotherapeutic agent, radiation therapy, or anesthetic.

Formulations of the food composition of the present invention may be prepared by conventional methods and may be formulated into capsules or other tablets, granules, powders, beverages, pills, etc. after drying with the carrier, It can be manufactured in food form including functional food.

The content of the active ingredient in the composition of the present invention may be contained in an amount of 0.01 to 20% based on the total weight of the composition.

The food composition of the present invention can also be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles for the purpose of preventing and / or improving vomiting or diarrhea.

For example, the health functional food of the tablet form may be prepared by granulating a mixture of a tofu extract with an excipient, a binder, a disintegrant, and other additives by a conventional method, Can be directly compression molded. The health functional food of the tablet form may contain a mating agent and the like if necessary, and may be sieved to a suitable skin care agent if necessary.

The matters mentioned in the uses, compositions and methods of the present invention are applied equally as far as they are not mutually contradictory.

The composition comprising the tofu extract of the present invention as an active ingredient exhibits prevention, improvement, and / or therapeutic effect of vomiting or diarrhea. Especially vomiting or diarrhea suppressed by 5-HT ₃ receptor selective antagonists without adverse effects caused by acting on dopamine receptors, such as chemotherapeutic agents, radiation treatment, or prevention and improvement of vomiting or diarrhea due to awakening after anesthetic administration, and / ≪ / RTI >

Figure 1 shows the concentration-response relationship from on NG (ganglion ganglion) neurons to ondansetron.
Figure 2 shows the effect of Machili Thunbergii Cortex Extract on 5-HT induced current in NG neurons.

Examples and production examples are provided to facilitate understanding of the present invention. The following examples and preparation examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples and the production examples.

Example  One. Too late  Preparation of extract

The ethanol extracts of tobacco leaves were prepared by the following method. To 100 g of the soymilk, 130% of ethyl alcohol was added to the volume of the soymilk, and sonication was performed at room temperature for 15 minutes for 10 days at room temperature for 3 days. The extracted solvent was concentrated under reduced pressure with a rotay evaporator and lyophilized to obtain 9.88 g of an extract. The extracts used were dissolved in DMSO (final concentration <0.3%) at a concentration of 0.2 mg / ml.

Experimental Example  One. Too late  The 5- HT  Confirming induction current suppression effect

1-1. Preparation of sample

The tobacco extract prepared in Example 1 was used for the experiment.

5-HT (serotonin) (Sigma-Aldrich) was dissolved in distilled water (DW) to prepare a 10 mM stock solution and diluted to 3 × 10 ^-6 M in the experiment to induce inward current.

The 5-HT ₃ receptor antagonist ondansetron was a Tocris Cookson product.

1-2. Isolation of vagus nerve cells

The experimental animals were Sprague-Dawley rat female rats, 10 to 14 days old, who were ordered from Orient Bio. Rats were fed with feed and water in a feeding room where temperature, humidity, and illumination (12 hours / cancer) were maintained automatically.

After rats were anesthetized (Zolazepam 0.3 mg / kg), the central part of the neck was opened, the vagus nerve was exposed, and a portion of the nodose ganglion (NG) was removed. The ACSF solution (composition mM: 125 NaCl, 3 KCl, 1.2 KH ₂ PO ₄ , 1.2 MgSO ₄ , 25 NaHCO ₃ , 10 glucose, 2 CaCl ₂ ). The ACSF solution was bubbled with 95% O ₂ - 5% CO ₂ gas for 20 minutes at low temperature before use. After removing the membranes of the ganglion ganglia, they were placed in an ACSF solution containing type Ⅱ trypsin (5 mg / ml at 1310 units / mg; Sigma-Aldrich., St. Louis, Mo.) Lt; / RTI &gt; The cells were then placed in ACSF and stored at room temperature for 1-3 hours. The tissues stored in the ACSF were placed in a chamber of standard external recording solution (composition mM: 150 NaCl, 5 KCl, 1 MgCl ₂ , 2 CaCl ₂ , 10 HEPES, 10 glucose, adjusted to pH 7.4 with Trisbase) The neurons were separated by pipetting with a glass pipette and used for electrophysiological recording.

1-3. Vagus nerve cell Ondansetron  or Too late  Extract treatment

(One) Ondansetron  process

To investigate the effect of ontansetron, a selective antagonist of 5-HT ₃ receptor, on 5-HT induced current in vagal nerve cells, ondansetron was added to the vaginal nerve cells prepared in 1-2 above (10 ^-12 , 10 ^-11 , 10 ^-10 , 10 ^-9 , 10 ^-8 M) were pretreated for 30 seconds and then used with 3x10 ^-6 M 5-HT.

(2) Too late  Extract treatment

In order to observe the effect of the soymilk on the 5-HT induced current in the vagus nerve cells, the tobacco ethanol extract was pretreated for 30 seconds at the concentration of 0.02 mg / ml and 0.2 mg / ml, respectively, And then treated with 3x10 ^-6 M 5-HT.

1-4. In vagal nerve cells, 5- HT  Inductive current measurement: electrophysiological record

In this experiment, the 5-HT reaction was recorded by voltage clamp method using Multiclamp 700B (Molecular Devices, Sunnyvale, California) and pClamp 9 software (Molecular Devices). Electrophoresis was carried out by punching with nystatin at a concentration of 450 ug / ml. The glass electrode used for the recording was filled with an internal solution (pH 7.2) composed of 50 mM KCl, 100 mM K-gluconate and 10 mM HEPES. Single cell fixed membrane voltage was set to -50 mV and current was filtered at 3 KHz and sampled at 10 KHz and stored in a computer. All experiments were conducted at room temperature (21-22 ° C).

All data were analyzed using pClamp 9 program and expressed as mean ± SE. Statistical analysis was performed using a one-way ANOVA with post-hoc multiple comparisons (Bonferrorni / Dunn's correction, StatView, SAS Institute Inc., Cary, NC) .

1-5. Experiment result

(1) In vagus nerve cells Ondansetron  5- HT  Effect on induced current

The results of the 5-HT induced current measurement in the vagus nerve cells upon ondansetron treatment are shown in FIG. Figure 1 shows the concentration-response relationship from on NG (ganglion ganglion) neurons to ondansetron. In FIG. 1 A, the traces represent 3x10 ^-6 M 5-HT induced currents by ondansetron. All traces were obtained from the same neurons. Figure 1B shows the concentration-inhibition relationship of the ondansetron. Each point is an average of 7 neurons.

As shown in FIG. 1, ondansetron at a concentration of 10 ^-12 M to 10 ^-8 M inhibited the 5-HT induction current in a concentration-dependent manner and completely inhibited it at 10 ^-8 M (p <0.0001). Therefore, 5-HT induced currents formed in vagal nerve cells could be estimated to be generated through 5-HT ₃ receptors.

After washing for 5 min, the 5-HT induction current returned to control level again.

(2) In vagal nerve cells Too late  When the extract is 5- HT  Effect on induced current

The results of the 5-HT induced current measurement in vagal nerve cells in the treatment of tobacco extract are shown in Fig. FIG. 2 shows the effect of the tobacco extract on the 5-HT induced current in NG neurons. In FIG. 2A, the traces show inhibition of the 5-HT induced current by the tobacco extract (0.2 mg / ml). In Figure 2B, each column represents the mean ± SE value from four neurons. *** p <0.05, ** p <0.001, significant compared to the control group.

As shown in Fig. 2, it was observed that tobacco extract completely inhibited 5-HT induced current at 0.2 mg / ml (p < 0.001). Therefore, it is thought that tobacco suppresses vomiting or diarrhea suppression by inhibiting the binding of 5-HT ₃ receptor and 5-HT.

Claims (6)

A pharmaceutical composition for preventing or treating vomiting caused by administration of a chemotherapeutic agent, radiation treatment, or anesthetic, which contains an extract of Machili Thunbergii Cortex as an active ingredient. The composition of claim 1, wherein the tofu extract exhibits a selective antagonistic effect on the 5-HT ₃ receptor. delete A food composition for prevention or amelioration of vomiting caused by administration of a chemotherapeutic agent, radiation treatment, or anesthetic, which comprises a tobacco extract as an active ingredient. 5. The composition of claim 4, wherein the tofu extract exhibits a selective antagonistic effect on the 5-HT ₃ receptor. delete

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