KR102282535B1 - Pharmaceutical composition for preventing or treating of irradiation-induced cell or tissue damage - Google Patents

Abstract

Disclosed is a pharmaceutical composition for the prevention or treatment of tissue or cell damage caused by radiation, wherein the pharmaceutical composition increases the survival rate of irradiated mice, damage recovery effect of hematopoietic cells (blood cells and bone marrow cells), small intestine villi A pharmaceutical composition for prevention, alleviation or treatment of cell or tissue damage caused by radiation exposure, cell or tissue damage caused by radiation exposure, because it has the effect of reducing the length and restoring the reduction in the circumference of the seminiferous tubule of the testis testis, and blocking apoptosis. It can be usefully used as a health functional food composition for prevention, alleviation or improvement of

Description

Pharmaceutical composition for the prevention or treatment of tissue or cell damage caused by irradiation TECHNICAL FIELD [0002] Pharmaceutical composition for preventing or treating of irradiation-induced cell or tissue damage

It relates to a pharmaceutical composition for preventing or treating tissue or cell damage caused by radiation.

There are various cases of damage to the living body due to radiation exposure, such as radiation exposure accidents at industrial sites dealing with radiation and side effects on normal tissues that occur during radiation treatment. In particular, after the 9/11 incident in the United States in 2001 and the Fukushima nuclear accident in Japan in 2011, government-led research has been intensively carried out as a countermeasure for large-scale radiation accidents. After 9/11, the U.S. Department of Defense's Armed Forces Radiobiology Research Institute (AFRRI) has been actively conducting research on overcoming biological side effects and biological defense mechanisms caused by radiation, and the National Institute of Allergy and Infectious Diseases (NIAID) of the US National Institutes of Health The Centers for Countermeasures Against Radiation (CMCR) was established to invest heavily in radiation and nuclear medical countermeasures programs. In addition, the current 'Radiation Safety Management Act' has been implemented in Korea amid growing anxiety about radiation exposure to prevent unnecessary exposure to radiation that may occur in the surroundings.

Radiation is widely used for the purpose of treating cancer in various tissues and prolonging the life of cancer patients by removing or inhibiting the growth of cancer cells, but it can cause damage to the normal tissues of long-term cancer survivors and develop into secondary cancer. (Dormand et al. 2005). Radiation causes direct tissue damage and indirect damage due to ionization of biological materials, and reactive oxygen species (ROS) generated by radiation indirectly causes cell damage and apoptosis such as DNA, proteins, and cell membranes. induces (Halliwell and Gutterige 1999).

In addition, when irradiated with radiation, there is a problem that organs such as liver and kidneys can be greatly damaged by MDA (malondialdehyde) formation and protein modification by lipid peroxidation (Garner A et al., Int. J. Radiat. Biol). (Relat. Stud. Phys. Chem. Med., 50,323-335, 1986).

Therefore, research is being conducted to develop a new efficient medical technology for measuring, diagnosing, or treating damage during radiation exposure, and to use it in a technology to improve treatment efficiency through protection of normal tissue damage during radiation treatment. are radioprotectants applied prior to radiation exposure; radiomitigators applied during radiation exposure or before the onset of obvious signs within a short period of time after exposure; and radiotherapy agents that are applied after clear signs appear by radiation exposure.

A representative example of a radioprotective agent is Amifostine (Ethyol R), which is an aminothiols derivative. Aminothiols are radioprotective agents that act as free radical scavengers as chemical analogues of cysteine. Among them, amifostine (WR-2721), known in 1969, was developed by The Walter Reed Army Institute of Research program, and more than 4,000 aminothiol derivative compounds have been developed and tested to date.

Amifostine is used in tumor radiation therapy because it protects normal tissues from radiation and does not protect solid tumors. In addition, as a treatment for dry mouth, the most frequently occurring side effect of radiation therapy in the treatment of head and neck cancer, amifostine is the first drug approved by the FDA. Currently, it is used for many patients after chemotherapy, but the efficacy due to the antioxidant effect is weak, and side effects such as nausea, vomiting, and hypotension appear, so it is used limitedly.

One aspect of the present invention is to provide a pharmaceutical composition for preventing, alleviating or treating cell or tissue damage caused by radiation exposure.

Another aspect of the present invention is to provide a health functional food composition for preventing, alleviating or improving cell or tissue damage caused by radiation exposure.

In order to achieve the above object,

In one aspect of the present invention, a compound represented by the following formula (1), a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof, as an active ingredient, prevention of cell or tissue damage caused by radiation exposure , to provide a pharmaceutical composition for alleviation or treatment.

[Formula 1]

In Formula 1,

R ¹ , R ² , R ³ , and R ⁴ are as defined herein.

Another aspect of the present invention provides a method of protecting or alleviating radiation from damage to cells or tissues caused by radiation exposure, comprising administering a therapeutically effective amount of the pharmaceutical composition to an animal other than a human.

Another aspect of the present invention is to prevent damage to cells or tissues by radiation exposure containing a compound represented by the following formula (1), a stereoisomer, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient , to provide a health functional food composition for alleviation or improvement.

[Formula 1]

In Formula 1,

R ¹ , R ² , R ³ , and R ⁴ are as defined herein.

Another aspect of the present invention provides a method for preventing, alleviating or treating cell or tissue damage caused by radiation exposure, comprising administering the pharmaceutical composition or health functional food composition to a subject in need.

Another aspect of the present invention provides the use of the pharmaceutical composition or health functional food composition in the prevention, alleviation or treatment of cell or tissue damage caused by radiation exposure.

The pharmaceutical composition of the present invention increases the survival rate of the irradiated mice, the effect of recovery from damage to hematopoietic cells (blood cells and bone marrow cells), the reduction in the length of the small intestine villi, the recovery of the reduction in the circumference of the seminiferous tubule of the testis testis, and apoptosis Since it has a blocking effect, it can be usefully used as a pharmaceutical composition for preventing, alleviating or treating cell or tissue damage caused by radiation exposure, and as a health functional food composition for preventing, alleviating or improving cell or tissue damage caused by radiation exposure.

1 is a graph showing the evaluation of the viability of mice when the pharmaceutical composition of Example 1 is administered to the radiation-irradiated mice performed in Experimental Example 1.
2 is a graph showing the analysis of changes in leukocytes, red blood cells, hemoglobin, hematocrit, and platelets of the mouse when the pharmaceutical composition of Example 1 is administered to the radiation-irradiated mice performed in Experimental Example 2.
Figure 3 is a graph showing the analysis of bone marrow cell changes in the mouse when the pharmaceutical composition of Example 1 is administered to the radiation-irradiated mice performed in Experimental Example 2.
4 is a graph showing the analysis of changes in the gastrointestinal tract of the mouse when the pharmaceutical composition of Example 1 is administered to the mice irradiated with radiation performed in Experimental Example 3;
5 is an image and graph showing the analysis of changes in the testis of the mouse testis when the pharmaceutical composition of Example 1 is administered to the radiation-irradiated mice performed in Experimental Example 3. FIG.
6 is a graph showing the analysis of changes in bone marrow apoptosis in mice when the pharmaceutical composition of Example 1 is administered to the radiation-irradiated mice performed in Experimental Example 4.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention is a compound represented by the following formula (1), a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof. Prevention of damage to cells or tissues caused by radiation exposure, Provided is a pharmaceutical composition for palliative or therapeutic use:

[Formula 1]

In Formula 1,

R ¹ is hydrogen, CN, NO ₂ or NH ₂ ;

R ² is —CH(OR ^a ) ₂ ;

R ³ and R ⁴ are independently hydrogen, halogen, straight-chain or branched C _1-5 alkyl unsubstituted or substituted one or more with halogen, straight-chain or branched C _{1 , unsubstituted or substituted with one or more halogen -4} alkoxy, NO ₂ , or —CO ₂ R ^a ; and

Wherein R ^a is straight-chain or branched C _1-5 alkyl.

In Formula 1,

wherein R ¹ is NH ₂ ;

R ² is —CH(OR ^a ) ₂ ;

R ³ and R ⁴ are independently hydrogen or halogen; and

Wherein R ^a may be a straight-chain or branched C _1-3 alkyl.

Preferred examples of the compound represented by the above formula (1) include a compound of the following formula (A):

[Formula A]

.

.

The radiation is any one selected from the group consisting of UV, X-rays, alpha-rays, beta-rays, gamma rays, electron beams and neutron rays, and the experiment was performed using X-rays in the following experimental examples, but is not limited thereto.

In addition, the cell or tissue is not particularly limited, but refers to cells or tissues of the hematopoietic system, the gastrointestinal system, and the reproductive system. The hematopoietic tissue refers to a tissue related to the production of blood cells, specific examples thereof include bone marrow or spleen, and specific examples of the hematopoietic cells include blood cells or bone marrow cells. In addition, the gastrointestinal tissue represents the stomach, small intestine, and large intestine organs, and in the following experimental example, an experiment was performed using the villi of the ileum in the small intestine, but is not limited thereto, and the reproductive system tissue is the uterus, testis, ovary or Reproductive organs such as testis are shown, and in the following experimental examples, an experiment was performed using testis of a testis, but the present invention is not limited thereto.

The pharmaceutical composition of the present invention restores a decrease in hematopoietic cells of blood cells or bone marrow cells caused by radiation exposure, and restores organ damage caused by radiation exposure.

Furthermore, the radiation exposure may be due to radiation treatment or work in a radiation-related hazardous area, but is not limited thereto.

The radiation therapy may be radiation therapy for cancer, but is not limited thereto.

The pharmaceutical composition may be administered before or after radiation exposure.

The pharmaceutical composition may be prepared and used as an oral formulation or an injection formulation.

As a result of evaluating the survival rate of irradiated mice, recovery of damage to organs (gastrointestinal organs, reproductive organs), recovery of damage to hematopoietic tissue, and inhibition of apoptosis of irradiated cells,

The pharmaceutical composition has an effect of increasing the survival rate of irradiated mice, restoring a decrease in the length of small intestine villi, restoration of a decrease in the circumferential length of the seminiferous tubules of the testes, and recovery of damage to hematopoietic cells (blood cells and bone marrow cells), It can be seen that this blocks apoptosis of the irradiated cells (see Experimental Examples 1 to 4 and FIGS. 1 to 6).

Therefore, the pharmaceutical composition according to one aspect of the present invention can be usefully used as a pharmaceutical composition for preventing, alleviating or treating cell or tissue damage caused by radiation exposure.

In addition, since the compound represented by Formula 1 exhibits the same effects as described above, it may be usefully used as a composition for radiation protection.

The compound represented by Formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids, etc., organic acids such as trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, etc. get it from Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, sube Late, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compound represented by Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. and adding an organic or inorganic acid. It can be prepared by filtration and drying the precipitate, or by distilling the solvent and excess acid under reduced pressure and then drying and crystallizing in an organic solvent.

In addition, bases can be used to prepare pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. In this case, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt. The corresponding salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg silver nitrate).

Furthermore, the present invention includes not only the compound represented by Formula 1 and pharmaceutically acceptable salts thereof, but also solvates, optical isomers, hydrates, and the like, which can be prepared therefrom.

The term “hydrate” refers to a compound of the present invention comprising a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolec μLar forces. or salts thereof. The hydrate of the compound represented by Formula 1 of the present invention may include a stoichiometric or non-stoichiometric amount of water that is bound by non-covalent intermolecular forces. The hydrate may contain 1 equivalent or more, preferably, 1 to 5 equivalents of water. Such a hydrate may be prepared by crystallizing the compound represented by Formula 1 of the present invention, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof from water or a solvent containing water.

The term “solvate” means a compound of the invention or a salt thereof which contains either a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. Preferred solvents therefor include solvents that are volatile, non-toxic, and/or suitable for administration to humans.

The term "isomer" refers to a compound of the present invention or a salt thereof having the same chemical formula or molecular formula but sterically different. Such isomers include isomers such as tautomers, R or S isomers having an asymmetric carbon center, stereoisomers such as geometric isomers (trans, cis), and optical isomers (enantiomers). All these stereoisomers and mixtures thereof are also included within the scope of the present invention.

A pharmaceutical composition comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient may be administered parenterally, and parenteral administration is administered by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection. depending on how

At this time, in order to formulate a formulation for parenteral administration, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is mixed with water together with a stabilizer or buffer to prepare a solution or suspension, which is an ampoule or vial unit dosage form. can be manufactured with The composition may be sterilized and/or contain adjuvants such as preservatives, stabilizing agents, wetting or emulsifying agents, salts and/or buffers for regulating osmotic pressure, and other therapeutically useful substances, and mixing, granulation, in a conventional manner. It can be formulated according to the method of formulation or coating.

Formulations for oral administration include, for example, tablets, pills, hard/soft capsules, solutions, suspensions, emulsifiers, syrups, granules, elixirs, troches, and the like. , dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine), lubricants (eg silica, talc, stearic acid and its magnesium or calcium salts and/or polyethylene glycol). Tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine and the like, and optionally boron such as starch, agar, alginic acid or its sodium salt. It may contain releasing or boiling mixtures and/or absorbents, colorants, flavoring agents, and sweetening agents.

In addition, another aspect of the present invention provides a health functional food composition for preventing, alleviating or improving cell or tissue damage caused by radiation exposure containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient do:

[Formula 1]

In Formula 1,

R ¹ is hydrogen, CN, NO ₂ or NH ₂ ;

R ² is —CH(OR ^a ) ₂ ;

R ³ and R ⁴ are independently hydrogen, halogen, straight-chain or branched C _1-5 alkyl unsubstituted or substituted one or more with halogen, straight-chain or branched C _{1 , unsubstituted or substituted with one or more halogen -4} alkoxy, NO ₂ , or —CO ₂ R ^a ; and

Wherein R ^a is straight-chain or branched C _1-5 alkyl.

In Formula 1,

wherein R ¹ is NH ₂ ;

R ² is —CH(OR ^a ) ₂ ;

R ³ and R ⁴ are independently hydrogen or halogen; and

Wherein R ^a may be a straight-chain or branched C _1-3 alkyl.

Preferred examples of the compound represented by the above formula (1) include a compound of the following formula (A):

[Formula A]

.

.

The health functional food composition has an effect of increasing the survival rate of irradiated mice, restoring a decrease in the length of small intestine villi, restoration of a decrease in the circumference of the seminiferous tubule of the testis and testis, and recovery of damage to hematopoietic cells (blood cells and bone marrow cells) It can be seen that the radiation blocks apoptosis of the irradiated cells (see Experimental Examples 1 to 4 and FIGS. 1 to 6).

Therefore, the health functional food composition according to one aspect of the present invention can be usefully used as a health functional food composition for preventing, alleviating or improving cell or tissue damage caused by radiation exposure.

As used herein, the term 'health functional food' refers to food prepared by adding the compound of Formula 1 to food materials such as beverages, teas, spices, gums, and confectionery, or encapsulating, powdering, suspension, etc. It means that it has a specific effect on health, but unlike general drugs, it has the advantage of not having side effects that may occur when taking a drug for a long period of time using food as a raw material. The health functional food according to one aspect of the present invention obtained in this way is very useful because it can be ingested on a daily basis, and thus a high skin whitening effect can be expected.

The compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may be added to food as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be suitably determined according to the purpose of its use (for prevention or improvement). In general, the amount of the compound in the health functional food may be added in an amount of 0.1 to 90 parts by weight based on the total weight of the food. However, in the case of long-term ingestion for health and hygiene purposes or for health control, the above amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

In addition, the health functional beverage composition according to an aspect of the present invention is not particularly limited in other ingredients except for containing the compound as an essential ingredient in the indicated ratio, and various flavoring agents or natural carbohydrates are added as an additional ingredient like a conventional beverage. can be contained as Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and manufactured flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 g of the composition of the present invention.

Furthermore, in addition to the above, the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as manufactured flavorants and natural flavorants, coloring agents, and thickeners (cheese, chocolate). etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages.

Another aspect of the present invention provides a method for preventing, alleviating or treating cell or tissue damage caused by radiation exposure, comprising administering the pharmaceutical composition or health functional food composition to a subject in need.

Another aspect of the present invention provides the use of the pharmaceutical composition or health functional food composition in the prevention, alleviation or treatment of cell or tissue damage caused by radiation exposure.

Hereinafter, it will be described in detail by way of Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

< Example 1 > Preparation of pharmaceutical compositions for prevention, alleviation or treatment of cell or tissue damage caused by radiation exposure

(2R,3R,4S)-6-amino-4-[n-(4-chlorophenyl)-N-(1H-imidazol-2ylmethyl)amino]- 3-hydroxyl-2-methyl-2-dimethoxymethyl-3,4-dihydro-2H-1-benzopyran was prepared.

[Formula A]

Purity: 95%

Molecular Weight: 458.94

The compound is a known material, and may be prepared by a known method, may be prepared by a method known in Korean Patent Registration No. 10-0492252 or Korean Patent Publication No. 10-2017-0061980, or may be purchased and used.

< Experimental example 1> Irradiated Mice viability (survival rate) impact assessment

When the pharmaceutical composition of Example 1 was administered to the irradiated mice, the following experiment was performed to evaluate the viability of the mice.

Mice (C57BL/6, male, 8 weeks old) used in this experiment were purchased from Orient Bio Inc. (Seongnam-si, Gyeonggi-do), temperature 22 ± 2 ℃ humidity 65 ± 5%, and contrast control at 12 hour intervals. In a controlled and clean animal room, drinking water and feed were freely available, and a acclimatization period of at least one week was passed. The animal experiment protocol and animal experiment protocol used in this experiment were approved and managed by the Animal Experiment Ethics Committee (IACUC) established at the Korea Research Institute of Chemical Technology.

Irradiation was conducted by irradiating 7 Gy of X-rays to the whole body of a 9-week-old mouse at a rate of 0.5 Gy per minute using an X-ray generator (X-Rad 320, Pricision X-Ray Inc., USA). Example 1 was dissolved in DMSO:EtOH:PEG400:Water=0.5:1.5:3:5 in amounts of 30 and 50 mg/kg/5ml, respectively, and intraperitoneally injected. The time of intraperitoneal injection was 24 hours before and 30 minutes before irradiation, a total of two intraperitoneal injections were performed. In the negative control group, 5 ml/kg of each solvent was intraperitoneally injected. The viability of mice was measured by mortality for 30 days after irradiation, and using Sigma Stat (Jandel Co., San Rafael, CA, USA), Kaplan-Meier analysis and log-rank (Log-) rank test) was statistically processed, and the results are shown in FIG. 1 .

As shown in FIG. 1, the mice of the administration group of Example 1 had a concentration-dependently and statistically significant increase in survival rate compared to the control group (X-ray + Vehicle). Specifically, it can be seen that the survival rate is increased even when a small dose of 30 mg/kg is administered. In addition, it can be seen that the weight loss according to radiation is also reduced in a concentration-dependent manner.

<Experimental Example 2> Analysis of changes in bone marrow and blood cells of irradiated mice

Since the tissue or organ most sensitive to radiation is known as bone marrow and blood cells, in order to analyze changes in bone marrow and blood cells of the mouse when the pharmaceutical composition of the present invention is administered to an irradiated mouse, the following The same experiment was performed.

Experimental method was performed in the same manner as in Experimental Example 1, except that the amount of X-ray irradiation was reduced to 4 Gy, and Example 1 was intraperitoneally injected twice at an amount of 50 mg/kg/5ml (24 hours before and 30 minutes ago). To analyze changes in blood cells, to measure the total blood count before and after irradiation, 3 days before, 5 days, 10 days, and 15 days after X-ray irradiation, blood was drawn from the orbital venous plexus of each mouse using a microtube. collected. The collected blood was placed in an ethylenediaminetetraacetic acid (EDTA) tube and measured with a hemocytometer (URIT-3000 VET Plus, URIT Medical Electronic Co., China).

To analyze changes in bone marrow, after 15 days of X-ray irradiation, the mice were euthanized and the femur was isolated, and the bone marrow cells in the femur were washed with 10 ml of phosphate buffered saline and then washed with a cytometer (TC20). ™ Automated Cell Counter) was used to measure the number of bone marrow cells, and the results are shown in FIGS. 2 and 3 .

As shown in FIG. 2, the irradiated mice (positive control group, X-ray + Vehicle) significantly reduced leukocytes, red blood cells, hemoglobin, hematocrit, and platelets compared to mice not irradiated (negative control group, Vehicle). can confirm. However, it can be seen that the group administered with Example 1 (X-ray + Example 1) recovered statistically significantly compared to the positive control group (X-ray + Vehicle) on the 10th and 15th days after irradiation.

As shown in Figure 3, the irradiated mice (positive control group, X-ray + Vehicle) significantly decreased the number of bone marrow cells compared to the mice not irradiated with radiation (negative control group, Vehicle), Example 1 (X -ray + Example 1) compared with the positive control group (X-ray + Vehicle) administration group, it can be seen that the number of bone marrow cells is statistically significantly increased.

< Experimental example 3> Analysis of organ changes in irradiated mice

In order to analyze the changes in the organs of the mice when the pharmaceutical composition of Example 1 was administered to the irradiated mice, changes in the testes of the testes in the gastrointestinal tract and reproductive organs were analyzed through the following experiments.

In order to analyze the effect of Example 1 on the gastrointestinal tract and testis of the testis, the experiment was carried out in the same manner as in Experimental Example 1, but Example 1 was intraperitoneally injected twice at an amount of 50 mg/kg/5ml (24 hours ago). and 30 minutes ago). After 10 days of X-ray irradiation, mice were euthanized, and the ileum and testis of the small intestine were fixed in 10% neutral buffered formalin for 2 days. After the fixed specimen was cut into 4 μm thick and attached to a slide, changes in villi and seminiferous tubules were observed through hematoxylin and eosin staining. The stained ileum and testis were measured for the length and width of the villi through the Image-pro plus program. Statistical processing was analyzed using Sigma Stat (Jandel Co., San Rafael, CA, USA), one way ANOVA, and then a post-hoc test was performed through Dunnett test. The results are shown in FIGS. 4 and 5 .

As shown in Figure 4, the irradiated mice (X-ray + Vehicle administration group), compared to the mice not irradiated with radiation (negative control group), the length of the gastrointestinal villi was significantly reduced, Example 1 (X-ray + Example 1) It can be seen that the administration group has a statistically significant increase in villi length compared to the positive control group (X-ray + Vehicle).

As shown in Figure 5, the irradiated mice (X-ray + Vehicle administration group), compared to the mice not irradiated with radiation (negative control group), the circumference of the seminiferous tubule was significantly reduced, Example 1 (X- It can be seen that the ray + Example 1) administration group had a statistically significant increase in the circumferential length of the seminiferous tubule compared to the positive control group (X-ray + Vehicle).

< Experimental example 4> Analysis of changes in bone marrow cell death in irradiated mice

For apoptosis analysis, mice were sacrificed by cervical dislocation and femurs were extracted. The femur was disinfected with 70% alcohol, both ends were cut and bone marrow cells were extracted. Then, 10ug/ml of Hoechst 33342 (ThermoFisher scientific, Bartlesville, OK, USA), 1ug/ml of FITC Annexin V (BD Biosciences, San Jose, CA, USA) and 10ug/ml of Propidium iodide (BD Biosciences) 3 The degree of lack of cell membrane asymmetry due to apoptosis was analyzed by heavy staining and quantified using NuCleoCounter NC3000TM (Chemometec, Allerod, Denmark). The group not irradiated with radiation was used as a reference, and the results are shown in FIG. 6 .

As shown in FIG. 6 , compared with mice not irradiated with radiation (negative control, Vehicle), irradiated mice (positive control, X-ray + Vehicle) significantly increased the rate of bone marrow cell death. On the other hand, it can be seen that Example 1 (X-ray + Example 1) administration group has a statistically significant decrease in bone marrow cell death rate when compared to the positive control group (X-ray + Vehicle).

Therefore, the pharmaceutical composition of the present invention increases the survival rate of the irradiated mice, the effect of repairing damage to hematopoietic cells (blood cells and bone marrow cells), the reduction in the length of the small intestine villi, the recovery of the reduction in the circumference of the seminiferous tubules of the testis, Since it has a cell death blocking effect, it can be usefully used as a pharmaceutical composition for preventing, alleviating or treating damage to cells or tissues caused by radiation exposure, and as a health functional food composition for preventing, alleviating or improving cell or tissue damage caused by radiation exposure. there is.

<Formulation Example 1> Preparation of powder

2 g of a compound represented by Formula 1

1 g lactose

The above ingredients were mixed and filled in an airtight cloth to prepare a powder.

<Formulation Example 2> Preparation of tablets

100 mg of the compound represented by Formula 1

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional method for manufacturing tablets.

<Formulation Example 3> Preparation of capsules

100 mg of the compound represented by Formula 1

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, the capsules were prepared by filling in gelatin capsules according to a conventional manufacturing method of capsules.

<Formulation Example 4> Preparation of injection

100 mg of the compound represented by Formula 1

mannitol 180 mg

Na ₂ HPO ₄ ㆍ2H ₂ O 26 mg

distilled water 2974 mg

According to a conventional method for preparing injections, injections were prepared by containing the above ingredients in the indicated amounts.

<Formulation Example 5> Preparation of ointment

5 g of a compound represented by Formula 1

20 g cetyl palmitate

40 g cetanol

40 g stearyl alcohol

80 g myristane isopropyl

60 g polysorbate

1 g of propyl paraoxybenzoate

1 g of methyl paraoxybenzoate

Appropriate amount of phosphoric acid and purified water

According to a conventional method for preparing an ointment, an ointment was prepared by containing the above ingredients in the indicated amounts.

<Formulation Example 6> Preparation of health functional food

500 ng of the compound represented by Formula 1

appropriate amount of vitamin mixture

Vitamin A Acetate 70mg

Vitamin E 1.0mg

Vitamin 0.13mg

Vitamin B2 0.15mg

Vitamin B6 0.5mg

0.2mg vitamin B12

Vitamin C 10mg

Biotin 10mg

Nicotinamide 1.7mg

50mg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture appropriate amount

1.75 mg of ferrous sulfate

Zinc Oxide 0.82mg

Magnesium carbonate 25.3mg

Monobasic potassium phosphate 15mg

Dicalcium Phosphate 55mg

Potassium citrate 90mg

100mg calcium carbonate

Magnesium Chloride 24.8mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health functional food, but the composition is mixed in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method. Then, the granules can be prepared and used in the preparation of a health functional food composition according to a conventional method.

<Formulation Example 7> Preparation of health functional beverage

500 ng of the compound represented by Formula 1

citric acid 1000mg

100 g of oligosaccharides

Plum Concentrate 2g

1 g of taurine

Total 900ml by adding purified water

After mixing the above ingredients according to the usual health function beverage manufacturing method, after stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized container, sealed and sterilized, then refrigerated, and then stored in a health function Used to prepare beverage compositions.

Although the composition ratio is composed of ingredients suitable for relatively preference beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences, such as the demand class, the demanding country, and the use.

Claims (11)

A pharmaceutical for preventing, alleviating or treating damage to cells or tissues caused by radiation exposure containing a compound represented by the following formula (1), a stereoisomer thereof, a solvate thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient Composition:
[Formula 1]

In Formula 1,
R ¹ is hydrogen, CN, NO ₂ or NH ₂ ;
R ² is —CH(OR ^a ) ₂ ;
R ³ and R ⁴ are independently hydrogen, halogen, straight-chain or branched C _1-5 alkyl unsubstituted or substituted one or more with halogen, straight-chain or branched C _{1 , unsubstituted or substituted with one or more halogen -4} alkoxy, NO ₂ , or —CO ₂ R ^a ; and
Wherein R ^a is straight-chain or branched C _1-5 alkyl.
According to claim 1,
wherein R ¹ is NH ₂ ;
R ² is —CH(OR ^a ) ₂ ;
R ³ and R ⁴ are independently hydrogen or halogen; and
Wherein R ^a is straight-chain or branched C _1-3 alkyl,
A pharmaceutical composition for preventing, alleviating or treating damage to cells or tissues caused by radiation exposure.
According to claim 1,
The compound represented by the formula (1) is a compound of the following formula (A), a pharmaceutical composition for preventing, alleviating or treating damage to cells or tissues caused by radiation exposure:
[Formula A]

.
According to claim 1,
The radiation is UV, X-rays, alpha-rays, beta-rays, gamma rays, electron beams, and a pharmaceutical composition for preventing, alleviating or treating cell or tissue damage caused by radiation exposure, characterized in that one selected from the group consisting of neutron rays.
According to claim 1,
The pharmaceutical composition is a pharmaceutical composition for preventing, alleviating or treating damage to cells or tissues caused by radiation exposure, characterized in that it restores the reduction in hematopoietic cells of blood cells or bone marrow cells caused by radiation exposure.
According to claim 1,
The pharmaceutical composition is a pharmaceutical composition for preventing, alleviating or treating damage to cells or tissues caused by radiation exposure, characterized in that it restores organ damage caused by radiation exposure.
According to claim 1,
The radiation exposure is radiation treatment or a pharmaceutical composition for prevention, alleviation or treatment of cell or tissue damage caused by radiation exposure, characterized in that by working in a radiation-related hazardous area.
According to claim 1,
The pharmaceutical composition is a pharmaceutical composition for preventing, alleviating or treating cell or tissue damage caused by radiation exposure, characterized in that it is administered before or after radiation exposure.
According to claim 1,
The pharmaceutical composition is a pharmaceutical composition for preventing, alleviating or treating cell or tissue damage caused by radiation exposure, characterized in that the pharmaceutical composition is an oral formulation or an injection formulation.
A method of radiation protection or alleviation against cell or tissue damage caused by radiation exposure, comprising administering a therapeutically effective amount of the pharmaceutical composition of claim 1 to animals other than humans.
A health functional food composition for preventing, alleviating or improving cell or tissue damage caused by radiation exposure containing a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

In Formula 1,
R ¹ is hydrogen, CN, NO ₂ or NH ₂ ;
R ² is —CH(OR ^a ) ₂ ;
R ³ and R ⁴ are independently hydrogen, halogen, straight-chain or branched C _1-5 alkyl unsubstituted or substituted one or more with halogen, straight-chain or branched C _{1 , unsubstituted or substituted with one or more halogen -4} alkoxy, NO ₂ , or —CO ₂ R ^a ; and
Wherein R ^a is straight-chain or branched C _1-5 alkyl.

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