KR20140058734A - Pharmaceutical composition for treatment or prevention of side-effect of anticancer drug and stomach disease - Google Patents

Abstract

The present invention provides a pharmaceutical composition for treating or preventing stomach disease and/or side-effects of an anticancer drug which includes minimal effective dose of compound represented by chemical formula 1, pharmaceutically acceptable salt thereof, pro-drug, solvate, or isomer, pharmaceutically acceptable carrier, diluent, or excipient, or a combination thereof. R1, R2, R3, R4, R5, R6, R7, R8 and n are defined as in the present specification.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a pharmaceutical composition for the treatment or prevention of side effects caused by anticancer drugs and gastrointestinal diseases,

The present invention relates to a pharmaceutical composition having pharmacological effects superior in the side effects caused by anticancer drugs and the treatment and prevention of gastrointestinal diseases, wherein the pharmaceutical composition comprises a cisplatin-induced gastric ulcer, gastric ulcer, vomiting, gastric ulcer , And exhibit excellent effects in the treatment and prevention of ethanol induced gastric ulcer.

Most chemotherapeutics for chemotherapy kill rapidly dividing and proliferating cancer cells, but they are toxic to cancer cells, as well as to bone marrow, immune cells and intestinal epithelium, because they affect the DNA synthesis process of the dividing cells ( Gibson et al., 2002; Bauda et al., 2003). Thus, bone marrow toxicity is a major cause of death due to loss of hematopoiesis, anemia due to immune cell toxicity, immunodeficiency, and intestinal injury due to malnutrition, dehydration and increased morbidity for infection (Cascinu, 1995). In particular, cisplatin induces severe vomiting and intestinal damage (Gupta and Sharma, 1996; Ueda et al., 1998; Yamakuni et al., 2002, 2006; Jeong et al., 2005; It is well known that it induces renal toxicity, hepatotoxicity and immune system toxicity (Talmadge et al., 1994; Sugiyama et al., 1995a, 1995b; Ueda et al , 1998, Kobayashi et al., Lee et al., 2008).

Such loss of function due to renal damage results in increased kidney and related structures, atrophy of the kidney, changes in body fluid volume, electrolyte imbalance, metabolic acidosis, gas exchange disorder, impaired anti-infective function and accumulation of toxic toxins.

Recently, in order to inhibit the side effects of anticancer drugs, various studies have been conducted to find substances effective for the prevention and treatment of cisplatin-induced kidney damage, but there are no drugs approved for prevention and treatment of kidney damage .

Activation of the serotonin type 3 (5-hydroxytryptamine 3, 5-HTC) receptor, which is distributed in the gastrointestinal tract and central nervous system, has been known to be one of the main mechanisms of nausea and vomiting due to administration of anticancer drugs (Andrews et al., 1998; Kilpatrick et al. al., 1990; Tyers, 1991; Andrews and Davis, 1993). Generally, anticancer drugs induce serotonin-induced enterocromaffin cells by inducing intestinal damage (Minami et al., 1997) and enterochromaffin cells (Andrews and Davis, 1993). Serotonin induces vomiting and central nervous responses in vivo by activating the chemoreceptor trigger zone of postrema as well as vagus and sympathetic neurons (afferent neurons) (Clarke and Davison, 1978; Andrews et al., 1990)

Thus, selective 5-HT3 receptor antagonists have been shown to be effective in the treatment and prevention of atherosclerosis in animals (Andrews and Bhandari, 1993; Rudd and Naylor, 1996) and humans (Andrews et al., 1990; Einhorn et al., 1990; Aapro, 1991; Morrow et al., 1995 ) Were effective in preventing nausea and vomiting. 5-HTC receptor antagonists such as tropisetron have been used to suppress vomiting by chemotherapeutic agents such as cisplatin, cyclophosphamide, and doxorubicin (Bleiberg et al., 1992, 1993; Bruntsch et al. 1993; Lee et al., 1993; Mantovani et al., 1996). (Jovanovic-Micic et al., 1995; Nystakidou et al., 1998), cannabinoids (Abrahamov et < RTI ID = 0.0 > al 1995) and neukinin 1 (NK 1) receptor antagonists (Bountra et al., 1993; Tattersall et al., 1994; Beattie et al., 1995; Gardner et al., 1995, 1996; Watson et al., 1995; Harrison et al., 2001) is also known to be an effective antinociceptive agent (Jordan et al., 2007). However, 5-HTC receptor antagonists such as tropisetron are relatively effective for early vomiting and require frequent re-administration due to short duration (Aapro, 1991; Andrews 1994; Jeong et al., 2005).

In order to evaluate the efficacy of harbor soil, various animals such as ferret, dog, suncus and monkey have been used (Rudd et al., 1994; Rudd and Naylor, 1996; Nakayama et al., 2005). However, rodents, heaviest ruminants, and horse ruminants are not able to vomit because of strong sphincter (Jeong et al., 2005). In addition, since dogs are easily eaten only by overeating of feed, confusion arises in reproducing the vomiting response due to the administration of anticancer drugs, ferrets are the most widely used for the evaluation of antiglaucoma due to the difficulty in management of monkeys and suncus .

Digestive ulcers refer to damage to the area where the mucous membrane is immersed in hydrochloric acid (HCl) and pepsin, which is normally covered by mucin secreted by mucus cells Wallace and Granger, 1996; Neal, 2003). Therefore, erosions of gastric mucosa due to various causes such as factors promoting gastric acid secretion, factors weakening or depleting mucin layer against pepsin invasion, factors causing local circulation disorder, cell damage and inflammatory reaction (Wallace and Granger, 1996; Neal, 2003; Isobe et al., 2004; Byun et al., 2007). Therefore, gastric ulcers are caused by drugs such as non-steroidal anti-inflammatory drugs (NSAIDs) that inhibit cyclooxygenase (COX), which is the enzyme of prostaglandins (PGs), which strengthens the mucin layer and smoothes the local blood (Rao et al., 2004; Kim et al., 2005), alcohol (Cao et al., 2004; Rao et al., 2004; (Rao et al., 2004), and gastric acid secretion and retention (Cao et al., 2004; Rao et al., 2004).

Anti-gastric ulcers include proton-pump inhibitors, antacids, antagonists of histamine (H ₂ antagonists) that block acid secretion from parietal cells, PGs and their derivatives , And H. pylori (Wallace and Ganger, 1996; Neal, 2003). Most stomach ulcers were inhibited during gastric acid secretion, and gastric acid secretion was found to be an important factor in ulceration induction (Cao et al., 2004).

On the other hand, Dansam is widely used as a medicinal herb in Northeast Asia since ancient times, and it is well known that it has an excellent effect for prevention and treatment of various cardiovascular diseases. The inventors of the present application filed a patent application in Korean Patent Nos. 2003-0099556, 2003-0099557, 2003-0099657, 2003-0099658, 2004-0036195, 2004-0036197 , 2004-0050200, etc. have suggested that the main ingredients of Dansam are excellent drugs for treating obesity, diabetes and metabolic diseases. In particular, the metabolism of cryptotanshinone, 15,16-dihydrotanshinone, Tanshinone II-A, and Tanshinone I, Have been identified as a key component in the treatment of syndromic diseases.

The inventors of the present application have repeatedly carried out various studies and experiments and have found that β-lapachone {7,8-dihydro-2,2-dimethyl-2H-naphtho (2,3- b ) dihydropyran-7,8- {2,3,3-trimethyl-2,3,4,5-tetrahydro-naphtho (2,3- b ) dihydrofuran-6,7-dione}, a-dunnione {2,3,3- Naphthoquinone compounds such as 3,4,5-tetrahydro-naphtho (2,3- b ) dihydrofuran-6,7-dione, nocardinone A, nocardinone B, lantalucratin A, lantalucratin B, lantalucratin C, / RTI > and / or can be used for the treatment or prevention of gastrointestinal disorders.

β-lapachone is obtained from the lapacho tree (Tabebuia avellanedae) native to South America and dunnione and α-dunnione from the leaves of Streptocarpus dunnii native to South America. These natural tricyclic naphthoquinone derivatives have been widely used in South America for treating Chagas disease, a typical endemic disease in South America including anticancer drugs, and its effect is also known to be excellent. Particularly, their pharmacological activity as an anticancer agent began to be noticed in the western world, and as disclosed in US Pat. No. 5,969,163, these tricyclic naphtoquinone derivatives were actually developed as various anti-cancer agents by various research groups have.

However, in spite of various studies, it is not known that these naphthoquinone compounds have pharmacological efficacy for the treatment or prevention of anticancer side effect diseases and / or gastrointestinal diseases.

The present inventors have found that naphthoquinone compounds such as β-lapachone, dunnione, α-dunnione, nocardinone A, nocardinone B, lantalucratin A, lantalucratin B and lantalucratin C, Based on the similarity of the structures, the inventors investigated their side effects of anticancer drugs, pharmacological actions as a therapeutic and preventive agent for gastrointestinal diseases.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition comprising, as an active ingredient, a naphthoquinone-based compound which is effective for the treatment and prevention of an anti-cancer agent side effect disease and / or gastrointestinal disease.

The pharmaceutical composition for the treatment and prevention of side effects and / or gastrointestinal diseases according to the present invention comprises (a) a pharmacologically effective amount of a compound represented by the following formula (1), a pharmaceutically acceptable salt thereof, a prodrug, a solvent Cargo or isomer thereof, and (b) a pharmaceutically acceptable carrier, diluent, or excipient, or a combination thereof.

(1)

(One)

In this formula,

R ₁ and R ₂ are each independently hydrogen, halogen, alkoxy, hydroxy or lower alkyl of 1 to 6 carbon atoms;

R ₃ , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are each independently selected from the group consisting of hydrogen, hydroxy, alkyl of 1 to 20 carbon atoms, alkene or alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, Or two of these substituents may form a cyclic structure by mutual bonding, wherein the cyclic structure may be a saturated structure or a partially or fully unsaturated structure;

n is 0 or 1, and when n is 0, adjacent carbon atoms thereof form a cyclic structure by a direct bond.

In order to confirm the anticancer drug side effects and / or the gastrointestinal diseases by the compounds of the formula 1, various experiments were conducted as shown in the following Experimental Examples. As a result, the compounds of the formula 1 were induced by cisplatin And showed excellent effect on gastric damage, intestinal injury and vomiting, and excellent effect on gastric secretion suppression effect and ethanol-induced gastric mucosal hemorrhage.

Therefore, the pharmaceutical composition of the present invention comprising the compound of formula (1) as an active ingredient is expected to be able to treat and prevent gastric ulcer caused by various gastric ulcers and ethanol accompanied by gastric acid secretion, as well as an effective inhibitory function against cancer-causing diseases.

The term " pharmaceutically acceptable salt " means a formulation of a compound that does not cause serious irritation to the organism to which the compound is administered and does not impair the biological activity and properties of the compound. The pharmaceutical salts may be formed with acids which form non-toxic acid addition salts containing a pharmaceutically acceptable anion, for example inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid and the like, Organic carboxylic acids such as acetic acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid and salicinic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p- And acid addition salts formed by the same sulfonic acids and the like. For example, pharmaceutically acceptable carboxylic acid salts include metal salts or alkaline earth metal salts formed with lithium, sodium, potassium, calcium, magnesium and the like, amino acid salts such as lysine, arginine and guanidine, dicyclohexylamine, N Organic salts such as methyl-D-glucamine, tris (hydroxymethyl) methylamine, diethanolamine, choline and triethylamine, and the like. The compound of formula (I) according to the present invention may be converted into a salt thereof by a conventional method.

The term " prodrug " refers to a substance that is transformed into a parent drug in vivo. Prodrugs are often used because they are easier to administer than parent drugs. For example, they may achieve bioactivity by oral administration, whereas the parent drug may not. Prodrugs may also have improved solubility in pharmaceutical compositions over the parent drug. For example, a prodrug is an ester that facilitates the passage of a cell membrane, which is hydrolyzed to a carboxylic acid that is active by metabolism in a cell whose water solubility is once beneficial, Drug "). ≪ / RTI > Another example of a prodrug may be a short peptide (polyamino acid) that is attached to an acid group that is converted by metabolism so that the peptide reveals its active site.

The term " solvate " is intended to include a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces Quot; means a compound of the present invention or a salt thereof. Preferred solvents therefor are volatile, non-toxic, and / or solvents suitable for administration to humans, and when the solvent is water, it refers to hydrate.

The term " isomer " means a compound of the present invention or a salt thereof, which has the same chemical or molecular formula but is optically or sterically different.

Unless otherwise stated, the term " compound of formula I " is used in a concept that includes both the compound itself, its pharmaceutically acceptable salts, prodrugs, solvates and isomers.

The term " alkyl " means an aliphatic hydrocarbon group. In the present invention, the term "alkyl" refers to a "saturated alkyl" meaning that it does not contain any alkene or alkyne moieties and an "unsaturated alkyl" meaning that the alkyl includes at least one alkene or alkyne moiety. And the like. &Quot; Alkenene " moiety refers to a group in which at least two carbon atoms are composed of at least one carbon-carbon double bond, and " alkyne " moiety refers to a group in which at least two carbon atoms have at least one carbon- ≪ / RTI > The alkyl may be branched, straight-chain or cyclic, and may be substituted or unsubstituted.

The term " heterocycloalky " refers to a substituent in which the ring carbon is replaced by oxygen, nitrogen, sulfur, etc. and includes, for example, furan, thiophene, pyrrole, pyrroline, pyrrolidine, oxazole, thiazole, Imidazoline, imidazoline, imidazolidine, pyrazole, pyrazoline, pyrazolidine, isothiazole, triazole, thiadiazole, pyran, pyridine, piperidine, morpholine, thiomorpholine, pyridazine, Pyrimidine, pyrazine, piperazine, triazine, and the like, but are not limited thereto.

The term " aryl " refers to an aromatic substituent having at least one ring having a covalent pi electron system and containing carbocyclic aryl (e.g., phenyl) and a heterocyclic aryl group (e.g., pyridine) . The term includes monocyclic or fused ring polycyclic (i.e., rings that divide adjacent pairs of carbon atoms) groups.

The term " heteroaryl " means an aromatic group containing at least one heterocyclic ring.

Examples of the aryl or heteroaryl include, but are not limited to, phenyl, furan, pyran, pyridyl, pyrimidyl, triazyl and the like.

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ in formula (1) according to the present invention may be optionally substituted. Examples of such substituents include cycloalkyl, aryl, Aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halogen, carbonyl, thiocarbonyl, O-carbamyl, Carbamoyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulphonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato, iso One or more substituents individually and independently selected from thio, amino, thioanino, nitro, silyl, trihalomethanesulfonyl, amino including mono- and di-substituted amino groups, and protected derivatives thereof. have.

Preferable examples of the compounds of formula (1) may be compounds of the following formulas (2) and (3). Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are the same as defined in Formula (1).

The compound of formula (2) is a compound in which n is 0 and adjacent carbon atoms form a cyclic structure (furan ring) by a direct bond. Hereinafter, the compound may sometimes be referred to as a 'furan compound' or a 'furano-o-naphthoquinone derivative' .

(2)

(2)

The compound of formula (3) is a compound wherein n is 1. Hereinafter, the compound may sometimes be referred to as a 'pyran compound' or 'pyrano-o-naphthoquinone'.

(3)

(3)

In the above formula (1), R ₁ and R ₂ may particularly preferably be hydrogen.

Particularly preferred examples of the furan compounds of formula 2 include compounds of formula 2a wherein R ₁ , R ₂ and R ₄ are each hydrogen, or a compound of formula 2b wherein R ₁ , R ₂ and R ₆ are each hydrogen, .

(2a)

(2a)

(2b)

(2b)

Particularly preferred examples of the pyran compounds of Formula 3 include compounds of Formula 3a wherein R ₁ , R ₂ , R ₅ , R ₆ , R _7, and R ₈ are each hydrogen.

(3a)

(3a)

The term "pharmaceutical composition" refers to a mixture of other chemical components, such as a compound of Formula 1 and a diluent or carrier. The pharmaceutical composition facilitates administration of the compound into the organism. Various techniques exist for administering the compounds, including, but not limited to, oral, injectable, aerosol, parenteral, and topical administration. The pharmaceutical composition may be obtained by reacting acid compounds such as hydrochloric acid, bromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.

The " therapeutically effective amount " means that the amount of the compound administered will alleviate or reduce to some extent one or more symptoms of the disorder being treated, delay the onset of clinical markers or symptoms of the disease Quot; means the amount of active ingredient that is effective to effect the treatment. Thus, a pharmacologically effective amount can be any amount effective to (1) reverse the rate of progression of the disease, (2) to some extent inhibit further progression of the disease, and / or (3) (Preferably, eliminating) the degree of the effect. A pharmacologically effective amount can be determined empirically by testing compounds in known in vivo and in vitro model systems for diseases in need of treatment.

The " carrier " is defined as a compound that facilitates the addition of a compound into a cell or tissue. For example, dimethylsulfoxide (DMSO) is a commonly used carrier that facilitates the introduction of many organic compounds into cells or tissues of an organism.

The term " diluent " is defined as a compound that not only stabilizes the biologically active form of the compound of interest, but also dilutes in the water in which the compound is dissolved. Salts dissolved in buffer solutions are used as diluents in the art. A commonly used buffer solution is phosphate buffered saline, since it mimics the salt state of the human solution. Since buffer salts can control the pH of the solution at low concentrations, buffer diluents rarely modify the biological activity of the compounds.

The compounds used herein may be administered to a human patient either as such, or as a pharmaceutical composition mixed with other active ingredients, such as in a combination therapy, or with a suitable carrier neprotic agent. A description of the formulation and administration of compounds in this application can be found in " Remington ' s Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 18th edition, 1990 ".

In the pharmaceutical composition for oral administration according to the present invention, the active material may have a crystalline structure with a high degree of crystallinity or a crystalline structure with a low degree of crystallinity. Preferably, it has a crystal structure with a low degree of crystallinity, thereby solving the poor solubility problem of the compound of the formula (1) or (2), and further increasing the dissolution rate and the absorption rate in the body.

The " crystallinity " is a weight fraction of the crystalline portion of the crystalline compound as a whole. The crystallinity can be measured by a known method. For example, And the crystallinity can be determined by a measurement method using heat of fusion. The intensity distribution on the X-ray diffraction can be determined by diffraction by the amorphous portion and diffraction by the crystalline portion , Or the infrared ray method obtained from the peak of the width between the crystalline bands of the infrared absorption spectrum can be used to measure the degree of crystallization.

In the pharmaceutical composition for oral administration according to the present invention, the crystallinity of the active material is preferably 50% or less, more preferably the amorphous crystal structure in which the inherent crystallinity of the material is completely lost. The amorphous compound exhibits a relatively high solubility as compared with the crystalline compound, and the dissolution rate and the absorption rate in the body can be significantly improved.

In one preferred example, the amorphous structure can be made in the course of making the active material into fine particles. The fine particles can be prepared, for example, by a spray drying method of an active material, a melting method of forming a polymer and a melt, a coprecipitation method of forming a coprecipitate with a polymer or the like by dissolving in a solvent, a body forming method, and solvent volatilization. Preferably, a spray drying method can be used. On the other hand, fine particle-forming of the active material by the mechanical pulverization method contributes to improvement of solubility due to a large specific surface area even if it is not an amorphous structure, that is, a crystalline crystal structure or a semi-crystalline crystal structure. As a result, Improvement can be achieved.

The spray drying method is a method of preparing microparticles by dissolving an active material in a predetermined solvent and drying the microparticles by spraying. In the spray drying process, the crystallinity of the naphthoquinone compound itself is considerably lost, .

The mechanical pulverization method is a method of pulverizing fine particles with a strong physical force applied to the active material particles, and may be a pulverizing process such as a jet mill, a ball mill, a vibration mill, a hammer mill, etc., Jet mills capable of performing milling are particularly preferred.

On the other hand, regardless of the crystal structure, as the particle size of the fine particle type active material decreases, the dissolution rate and solubility increase due to the increase of the specific surface area, but too small particle size is not easy to produce fine particles of such size In one preferred example, the particle size of the active material may be in the range of 5 nm to 500 [mu] m since agglomeration or aggregation may cause rather low solubility. In this range, the aggregation phenomenon is suppressed to the maximum, and the dissolution rate and solubility are maximized by the high specific surface area.

The compounds according to the present invention are useful for the treatment or prevention of side effects caused by anticancer drugs, in particular, renal toxicity, intestinal injury, and vomiting caused by cisplatin. Specifically, the present invention provides a novel treatment for the damage caused by the vicious cycle of the inflammatory reaction mediator caused by cisplatin and the immune cell-induced inflammatory response immersed in the kidney tissue, which excellently alleviates severe intestinal damage due to administration of cisplatin, And hepatotoxicity. It also alleviates cell loss in the bone marrow and spleen, restoring T and B-lymphocyte function.

It also shows excellent inhibitory effect on gastric acid secretion and ethanol induced gastric ulcer.

Therefore, the compound according to the present invention is thought to be effective for the protection and recovery of kidney, liver, immunity and hematopoietic function including intestinal tract as well as the inhibitory effect against cancer induced vomiting, and the improvement of gastric ulcer due to specific causes such as gastric hyperproliferation and hangover It is expected to be valid.

FIG. 1 is a graph showing the degree of kidney damage according to the treatment time of cisplatin using hematoxylin-eosin stain (H & E stain).
FIG. 2 is a diagram showing the protective effect of Compound 1 (? -Lapachone,? L) against cisplatin-induced renal injury through H & E staining.
FIG. 3 is a graph showing the inhibitory effect of Compound 1 on changes in cisplatin-induced renal perfusion tissue using PAS staining (periodic-acid-Schiff stain).
4 is a graph comparing the modulating effects of Compound 1 on the increase of serum creatinine and urea induced by cisplatin. * p < 0.05.
5 is a graph showing changes in the ratio of NAD + / NADH in the kidney tissues of cisplatin and Compound 1 treated group. * p &lt; 0.05.
6 is a graph showing Sirt1 expression in the kidney tissues of the cisplatin-treated group and the compound 1-treated group.
Fig. 7 is a chart showing the degree of acetylation of p65, a target protein of Sirt1 in renal tissue, in cisplatin-treated group and Compound 1-treated group.
8 is a graph showing the expression of Sirt3 in the kidney tissues of the cisplatin-treated group and the compound 1-treated group.
9 is a chart showing the degree of acetylation of p53, which is a target protein of Sirt3 in the kidney tissues of the cisplatin-treated group and the compound 1-treated group.
FIG. 10 is a graph showing the concentrations of TNF-? In serum and urine in the cisplatin-treated group and the compound 1-treated group. * p &lt; 0.05.
11 is a view showing the expression of inflammatory cytokines in the kidney tissue of the cisplatin-treated group and the compound 1-treated group.
Fig. 12 is a graph showing the expression of NF-kB p65 protein in the kidney tissue of the cisplatin-treated group and the compound 1-treated group.
13 is a graph showing the expression of NOX 1 in the kidney tissue of the cisplatin-treated group and the compound 1-treated group.
14 is a graph showing the expression of NOX 4 in the kidney tissues of the cisplatin-treated group and the compound 1-treated group.
15 is a view showing TLR4 protein expression in the kidney tissues of the cisplatin-treated group and the compound 1-treated group.
16 is a graph showing the expression of MCP-1 protein in renal tissue of cisplatin-treated group and compound 1-treated group.
FIG. 17 is a graph showing the weight change of rats after cisplatin (3.5 mg / kg) peritoneal administration for 4 to 7 days and oral administration of Compound 2 for 10 days (control group; cisplatin alone group; mg / kg Compound 2; cisplatin + 10 mg / kg Compound 2; cisplatin + 25 mg / kg Compound 2; cisplatin + 50 mg / kg Compound 2).
18 is a chart showing the results of observing the small intestine after oral administration of cisplatin 3.5 mg / kg and compound 2 into the peritoneal cavity of mice for 4 to 7 days.
FIG. 19 is a chart showing the observation results of the small intestine after oral administration of cisplatin 3.5 mg / kg and compound 2 into the peritoneal cavity of rats for 4 to 7 days.
FIG. 20 is a graph showing the results of observation of bone marrow after oral administration of cisplatin 3.5 mg / kg and compound 2 into the peritoneal cavity of rats for 4 to 7 days.
FIG. 21 is a graph showing the number of spleen cells after administration of 3.5 mg / kg of cisplatin into the abdominal cavity of rats for 4 to 7 days and orally administering Compound 2 (5-50 mg / kg) for 10 days.
FIG. 22 is a chart showing concanavalin A (ConA) response of T lymphocytes after oral administration of Compound 2 (5-50 mg / kg) to rats receiving intraperitoneal cisplatin (3.5 mg / kg) for 10 days.
23 is a graph showing the lipopolysaccharide (LPS) response of T lymphocytes after oral administration of Compound 2 (5-50 mg / kg) to rats receiving intraperitoneal cisplatin (3.5 mg / kg) for 10 days
24 is a diagram showing the effect of repeatedly orally administering Compound 2 (150 mg / kg) for 7 days to nausea and vomiting reactions induced by subcutaneous administration of cisplatin (10 mg / kg).
FIG. 25 is a graph showing the weight change of ferrets according to intraperitoneal administration of cisplatin (5 mg / kg) for 7 days and oral administration of compound 2 (cisplatin, cisplatin + 25 mg / kg compound 2, cisplatin + 50 mg / kg Compound 2; cisplatin + 100 mg / kg Compound 2).
Figure 26 is a representative capture result of nausea and vomiting reactions of ferrets (the left side shows nausea and the right side shows vomiting)
Figure 27 shows the results of oral administration of Compound 2 (25-100 mg / kg) for 7 days in nausea and vomiting reactions induced by intraperitoneal administration of cisplatin.
FIG. 28 is a graph showing the effect of the compound 1 on the amount of gastric juice secreted by the 6-hour pyloric ligation. FIG.
29 is a diagram showing the pH effect of gastric juice of Compound 1 according to 6-hour pyloric ligation,
30 is a graph showing the effect of the free hydrochloric acid amount of the gastric juice of Compound 1 upon 6-hour pyloric ligation.
31 is a graph showing the effect of the total acidity effect of the gastric juice of Compound 1 upon 6-hour pyloric ligation.
32 is a graph showing the effect of the compound 2 on the gastric juice secretion amount according to the 6-hour pyloric ligation.
33 is a graph showing the pH effect of gastric juice of Compound 2 according to 6-hour pyloric ligation,
34 is a graph showing the effect of the free hydrochloric acid amount of the gastric juice of Compound 2 according to 6-hour pyloric ligation.
35 is a graph showing the effect of the total acidity effect of the gastric juice of Compound 2 upon 6-hour pyloric ligation.
Figure 36 shows the results of stomach ulcer induced by ethanol (3 ml / kg) (compound upper left, compound 2 at the upper right 10 mg / kg, compound 2 at the lower left 30 mg / kg, / kg of pantoprazole).
Figure 37 is a graph showing compound 2 (black) and pantoprazole (gray) effect of gastric ulcer induced by ethanol (3 ml / kg).

In the pharmaceutical composition according to the present invention, the compounds of formula (1) can be prepared by various methods based on known methods and / or techniques of organic synthesis, as will be described later, It is to be understood that the present invention is not limited to the above embodiments, but other methods may be present.

Production method 1: Synthesis of lapachol derivatives and acid catalyzed cyclization

β-lapachone is obtained in a relatively small amount in lapacho trees, whereas lapachol, which is a raw material for β-lapachone synthesis, is obtained in a large amount in lapacho trees. Therefore, a method of synthesizing β-lapachone using lapachol has been developed long ago . That is, L. F. Fieser et al. Am. Chem. Scoc. 49 (1927), 857}, lapachol and sulfuric acid are mixed together and vigorously stirred at room temperature to obtain β-lapachone in a relatively good yield. In general, relatively simple tricyclic naphthoquinone (pyrano-o-naphthoquinone and furano-o-naphthoquinone) derivatives are synthesized in a relatively good yield through cyclization using sulfuric acid as a catalyst, To synthesize various compounds of formula (1).

Lapachol β-lapachone

These processes are summarized by more general chemical reaction equations as follows.

That is, when 2-hydroxy-1,4-naphthoquinone is reacted with various allylic bromides or its equivalents in the presence of a base, C-alkylation (C-alkylation) and O-alkylation (O- Depending on the reaction conditions, it is also possible to synthesize only one derivative. Where the O-alkylated derivative is converted to another type of C-alkylated derivative through a Claisen rearrangement reaction by refluxing with a solvent such as toluene or xylene, so that various types of 3-substituted-2-hydroxy- 4-naphthoquinone derivatives can be obtained. The various types of C-alkylated derivatives thus obtained can be used to synthesize pyrano-o-naphthoquinone or furano-o-naphthoquinone derivatives of the compounds of formula 1 by inducing a cyclization reaction using sulfuric acid as a catalyst.

Production method 2: Diels-Alder reaction using 3-methylene-1,2,4- [3H] naphthalenetrione

V. Nair et al. {Tetrahedron Lett. (3H) naphthalenetrione, which is produced by heating 2-hydroxy-1,4-naphthoquinone with formaldehyde, is reacted with various olefins It has been reported that various pyrano-o-naphthoquinone derivatives can be synthesized relatively easily by inducing Diels-Alder reaction with a compound. This method is advantageous in that it can synthesize various forms of pyrano-o-naphtho-quinone derivatives relatively easily compared to the reaction to induce the cyclization reaction of lapachol derivatives under sulfuric acid catalyst conditions.

Production method 3: Haloakylation and cyclization reaction by radical reaction

Methods used for the synthesis of cryptotanshinone, 15,16-dihydrotanshinone, etc., can also be conveniently used to synthesize furano-o-naphthoquinone derivatives. As reported by AC Baillie et al. (J. Chem. Soc. (C) 1968, 48-52), 2-haloethyl or 3-haloethyl radical species derived from 3-halopropanoic acid or 4-halobutanoic acid derivatives ? (2-haloethyl or 3-halopropyl) -2-hydroxy-1,4-naphthoquinone can be synthesized by reacting with 2-hydroxy-1,4-naphthoquinone. To produce various pyrano-o-naphthoquinone or furano-o-naphthoquinone derivatives.

Production method 4: cyclization reaction of 4,5-benzofurandione by Diels-Alder reaction

Another method that has been used for the synthesis of cryptotanshinone and 15,16-dihydrotanshinone is JK Snyder et al. (Tetrahedron Letters 28 (1987), 3427-3430) There is a way to notice this. This method can be used to synthesize furano-o-naphthoquinone derivatives by inducing Cycloaddition by Diels-Alder reaction between 4,5-benzofurandione derivatives and various diene derivatives.

In addition, various derivatives can be synthesized on the basis of the above-described methods by using appropriate synthetic methods depending on the kinds of substituents. Specific examples thereof are shown in Table 1 below. Specific production methods for these are described in the following examples.

One

C ₁₅ H ₁₄ O ₃ 242.27 Method 1 2

C ₁₅ H ₁₄ O ₃ 242.27 Method 1 3

C ₁₅ H ₁₄ O ₃ 242.27 Method 1 4

C ₁₄ H ₁₂ O ₃ 228.24 Method 1 5

C ₁₃ H ₁₀ O ₃ 214.22 Method 1 6

C ₁₂ H ₈ O ₃ 200.19 Method 2 7

C ₁₉ H ₁₄ O ₃ 290.31 Method 1 8

C ₁₉ H ₁₄ O ₃ 290.31 Method 1 9

C ₁₅ H ₁₂ O ₃ 240.25 Method 1 10

C ₁₆ H ₁₆ O ₄ 272.30 Method 1 11

C ₁₅ H ₁₂ O ₃ 240.25 Method 1 12

C ₁₆ H ₁₄ O ₃ 254.28 Method 2 13

C ₁₈ H ₁₈ O ₃ 282.33 Method 2 14

C ₂₁ H ₂₂ O ₃ 322.40 Method 2 15

C ₂₁ H ₂₂ O ₃ 322.40 Method 2 16

C ₁₄ H ₁₂ O ₃ 228.24 Method 1 17

C ₁₄ H ₁₂ O ₃ 228.24 Method 1 18

C ₁₄ H ₁₂ O ₃ 228.24 Method 1 19

C ₁₄ H ₁₂ O ₃ 228.24 Method 1 20

C ₂₀ H ₂₂ O ₃ 310.39 Method 1 21

C ₁₅ H ₁₃ ClO ₃ 276.71 Method 1 22

C ₁₆ H ₁₆ O ₃ 256.30 Method 1 23

C ₁₇ H ₁₈ O ₅ 302.32 Method 1 24

C ₁₆ H ₁₆ O ₃ 256.30 Method 1 25

C ₁₇ H ₁₈ O ₃ 270.32 Method 1 26

C ₂₀ H ₁₆ O ₃ 304.34 Method 1 27

C ₁₈ H ₁₈ O ₃ 282.33 Method 1 28

C ₁₇ H ₁₆ O ₃ 268.31 Method 1 29

C ₁₃ H ₈ O ₃ 212.20 Method 1 30

C ₁₃ H ₈ O ₃ 212.20 Method 4 31

C ₁₄ H ₁₀ O ₃ 226.23 Method 4 32

C ₁₄ H ₁₀ O ₃ 226.23 Method 4

The pharmaceutical compositions of the present invention can be prepared in a known manner, for example, by means of conventional mixing, dissolving, granulating, sugar-making, powdering, emulsifying, encapsulating, trapping and lyophilizing processes .

The pharmaceutical compositions for use according to the invention thus comprise one or more pharmacologically acceptable excipients which comprise excipients or auxiliaries which facilitate the treatment of the active compounds with pharmaceutically usable formulations Or may be prepared by a conventional method using a carrier. Suitable formulations depend on the route of administration chosen. Any of the known techniques, carriers and excipients may be used as appropriate and as understood in the art, for example, Remingston's Pharmaceutical Sciences, supra. In the present invention, the compound of Chemical Formula 1 may be formulated into injectable preparations and oral preparations according to the purpose.

For injection, the components of the present invention may be formulated as a liquid solution, preferably a pharmacologically compatible buffer such as Hank's solution, Ringer's solution, or physiological saline. For mucosal permeation administration, a non-permeabilizing agent suitable for the barrier to be passed is used in the formulation. Such impermeabilizers are generally known in the art.

For oral administration, the compounds can be readily formulated by combining the pharmacologically acceptable carriers known in the art with the active compounds. Such carriers allow the compounds of the present invention to be formulated into tablets, pills, powders, granules, sugars, capsules, liquids, gels, syrups, slurries, suspensions and the like. Preferably, capsules, tablets, pills, powders and granules are possible, and in particular, capsules and tablets are useful. Tablets and pills are preferably prepared as preservative. Pharmaceutical preparations for oral use can be prepared by mixing one or two or more compounds of the present invention with one or more excipients, optionally milling such mixture, and if necessary passing the appropriate adjuvant, then treating the mixture of granules Tablets or sugar cores can be obtained. Suitable excipients include, but are not limited to, fillers such as starch, starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, hydroxypropylmethyl- Carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP), and the like. If necessary, a carrier such as a crosslinking polyvinylpyrrolidone, a starch or a disintegrating agent such as alginic acid or sodium alginate and a lubricant such as magnesium stearate, a binder and the like may be added.

Pharmaceutical preparations that can be used orally may include soft seal capsules made of gelatin and a plasticizer such as glycol or sorbitol, as well as push-hold capsules made of gelatin. Capsules for push-fixing may also contain active ingredients, such as a filler such as lactose, a binder such as starch, and / or a mixture with talc or a lubricant such as magnesium stearate. In soft capsules, the active compounds may be dissolved or dispersed in suitable solvents such as fatty acids, liquid paraffin, or liquid polyethylene glycols. A stabilizer may also be included. All formulations for oral administration should be in amounts suitable for such administration.

Compounds may also be formulated for parenteral administration by injection, e. G., By large pill injection or continuous infusion. In addition, injectable formulations may be presented in unit dosage form as ampoules or multi-dose containers with preservatives added. The compositions may take such forms as suspensions, solutions, emulsions on oily or liquid vehicles, and may also contain components for the formulation, such as suspending, stabilizing and / or dispersing agents.

In addition, the active ingredient may be in the form of a powder for constitution with a suitable vehicle, such as sterile pyrogen-free water, before use.

Pharmaceutical compositions suitable for use in the present invention include compositions containing an effective amount of the active ingredients. More specifically, a therapeutically effective amount refers to an amount of a compound that is effective in prolonging the survival of an object to be treated or preventing, alleviating or alleviating the symptoms of the disease. The determination of a therapeutically effective amount is well within the ability of those skilled in the art, particularly in light of the detailed disclosure provided herein.

When formulated in unit dose form, the compound of formula (I) as an active ingredient is preferably contained in a unit dose of about 0.1 to 1,000 mg. The dosage of the compound of formula (1) depends on the physician's prescription depending on factors such as the patient's weight, age and the particular nature and severity of the disease. However, the dosage required for adult therapy is usually in the range of about 1 to 1000 mg per day, depending on the frequency and intensity of administration. A total dosage of about 1 to 500 mg per day, separated by a single dose when administered intramuscularly or intravenously to an adult, may be sufficient, but in some patients a higher daily dose may be desirable.

The present invention also provides a method of using a compound of formula (I) for the manufacture of a medicament for the treatment or prevention of an anti-cancer agent side effect disease and / or a gastrointestinal disorder. The term &quot; treatment &quot; of the disease syndrome refers to a disease or disorder which is caused by a disease or disorder when used in an object showing an onset symptom, such as kidney damage, intestinal injury and vomiting due to the administration of an anticancer drug, gastric ulcer accompanied by gastric acid secretion and gastric ulcer induced by ethanol, Quot; prevention &quot; refers to stopping or delaying the onset of symptoms when used in a high-risk object that does not show the onset symptoms.

The present invention will be described in detail with reference to the following examples and experimental examples, but the scope of the present invention is not limited thereto.

Example 1 Synthesis of β-Lapachone (Compound 1)

2-Hydroxy-1,4-naphthoquinone (17.4 g, 0.10 M) was dissolved in DMSO (120 mL) and LiH (0.88 g, 0.11 M) was added slowly. At this time, attention is required because hydrogen is generated. The reaction solution was stirred for 30 minutes while confirming that no more hydrogen was generated, and then Prenyl bromide (1-Bromo-3-methyl-2-butene) (15.9 g, 0.10 M) and LiI , 0.025M) was added slowly. The reaction solution was stirred vigorously for 12 hours while heated to 45 캜. The reaction solution was cooled to 10 ° C or lower, and then ice (76 g) was added thereto, followed by addition of water (250 ml), and then concentrated hydrochloric acid (25 ml) was slowly added thereto to maintain the pH of the acid at PH> 1. When the reaction mixture was stirred vigorously with EtOAc (200 mL), a white solid was formed which was insoluble in EtOAc. The solids were filtered off and the EtOAc layer was separated. The water layer was extracted one more time with EtOAc (100 mL) and combined with the organic layer extracted earlier. The organic layer was washed with 5% NaHCO ₃ (150 mL) and the organic layer was concentrated. The concentrate was dissolved in CH ₂ Cl ₂ (200 mL) and separated by shaking vigorously with 2N aqueous NaOH (70 mL). The CH ₂ Cl ₂ layer was treated with 2N aqueous NaOH (70 mL x 2) and separated two more times. When the separated aqueous solution is combined and adjusted to acidic pH> 2 using concentrated hydrochloric acid, a solid is formed. Lapachol was obtained by filtration and separation. The obtained Lapachol was recrystallized using 75% EtOH. Lapachol thus obtained was mixed with sulfuric acid (80 ml), stirred vigorously at room temperature for 10 minutes, and then the reaction was terminated by adding ice (200 g). CH ₂ Cl ₂ (60 mL) was added to the reaction mixture, and the mixture was shaken vigorously. The CH ₂ Cl ₂ layer was separated and washed with 5% NaHCO ₃ . The water layer was extracted one more time with CH ₂ Cl ₂ (30 mL), washed with 5% NaHCO ₃ , and combined with the organic layer extracted earlier. The organic layer was dried over MgSO ₄ and concentrated to obtain impure β-Lapachone. This was further recrystallized using isopropanol to obtain pure β-Lapachone (8.37 g).

¹ H-NMR (CDCl ₃ ,?): 8.05 (1H, dd, J = 1, 8 Hz), 7.82 (1H, dd, J = (2H, t, J = 6.5 Hz), 7.50 (1H, dt, J = 1,8 Hz), 2.57

Example 2: Synthesis of Dunnione (Compound 2)

In the process of obtaining lapachol in Example 1, the solids which are not dissolved in EtOAc are O-Akylated 2-Prenyloxy-1,4-maphthoquinone, unlike the C-Alylation material Lapachol. This was purified first by further recrystallization with EtOAc. The purified solid (3.65 g, 0.015 M) was dissolved in toluene and the toluene was refluxed for 5 hours to induce Claisen rearrangement. The toluene was concentrated by distillation under reduced pressure, and the mixture was vigorously stirred at room temperature for 10 minutes while being mixed with sulfuric acid (15 ml) without further purification, and then the reaction was terminated by adding ice (100 g). CH ₂ Cl ₂ (50 mL) was added to the reaction mixture, and the mixture was shaken vigorously. The CH ₂ Cl ₂ layer was separated and washed with 5% NaHCO ₃ . The aqueous layer was further extracted once more with CH ₂ Cl ₂ (20 mL), washed with 5% NaHCO ₃ , and combined with the organic layer extracted earlier. The organic layer was dried over MgSO ₄ , concentrated, and chromatographed using silica gel to obtain 2.32 g of Dunnione in a pure state.

_{^{1 H-NMR (CDCl 3,}} δ): 8.05 (1H, d, J = 8Hz), 7.64 (2H, d, J = 8Hz), 7.56 (1H, m), 4.67 (1H, q, J = 7Hz) , 1.47 (3H, d, J = 7 Hz), 1.45 (3H, s), 1.27 (3H, s)

Example 3: Synthesis of alpha-Dunnione (Compound 3)

By dissolving 2-Prenyloxy-1,4-maphthoquinone (4.8 g, 0.020 M) purified in Example 2 in xylene and refluxing the xylene for 15 hours, a much higher temperature condition and longer reaction conditions The Claisen Rearrangement was induced. In this process, α-Dunnione is obtained in which the cyclization reaction is proceeded with the Claisen rearrangement and the Lapachol derivative in which one of the two Methyl groups has migrated. The xylene was concentrated by distillation under reduced pressure and then chromatographed using silica gel to obtain alpha-Dunnione (1.65 g) in a pure state.

¹ H-NMR (CDCl ₃ ,?):? (1H, s), 8.06 (1H, d, J = 8 Hz), 7.64 (2H, m), 7.57 (3H, d, J = 7 Hz)

Example 4: Synthesis of Compound 4

2-Hydroxy-1,4-naphthoquinone (17.4 g, 0.10 M) was dissolved in DMSO (120 ml) and LiH (0.88 g, 0.11 M) was added slowly. At this time, attention was required because hydrogen was generated. The reaction solution was stirred for 30 minutes while confirming that no more hydrogen was generated. Then, Methallyl bromide (1-Bromo-2-methylpropene) (14.8 g, 0.11 M) and LiI (3.35 g, 0.025 M) Slowly. The reaction solution was stirred vigorously for 12 hours while heated to 45 캜. The reaction solution was cooled to 10 ° C or less, and then ice (80 g) was added thereto, followed by addition of water (250 ml), and then concentrated hydrochloric acid (25 ml) was slowly added to maintain the pH of the solution at pH> 1. CH ₂ Cl ₂ (200 mL) was added to the reaction mixture and the mixture was shaken vigorously. CH ₂ Cl ₂ (70 ml) was added to the water layer and extracted one more time, and combined with the previously separated organic layer. At this time, it was confirmed that two substances were newly formed in TLC, and they were used without any separation. The organic layer was concentrated by distillation and then refluxed in xylene for 8 hours. In this process, the two materials on TLC were combined to obtain a relatively pure Lapachol derivative. The lapachol derivative thus obtained was vigorously stirred at room temperature for 10 minutes in a mixed state with sulfuric acid (80 ml), and then the reaction was terminated by adding ice (200 g). CH ₂ Cl ₂ (80 mL) was added to the reaction mixture, and the mixture was shaken vigorously. The CH ₂ Cl ₂ layer was separated and washed with 5% NaHCO ₃ . The aqueous layer was further extracted once more with CH ₂ Cl ₂ (50 mL), washed with 5% NaHCO ₃ , and combined with the organic layer extracted earlier. The organic layer was dried using MgSO ₄ and concentrated to obtain an impure Lapachone derivative (Compound 4). This was recrystallized again using isopropanol to obtain Compound 4 (12.21 g) in a pure state.

_{^{1 H-NMR (CDCl 3,}} δ): 8.08 (1H, d, J = 8Hz), 7.64 (2H, m), 7.57 (1H, m), 2.95 (2H, s), 1.61 (6H, s)

Example 5: Synthesis of Compound 5

The reaction was carried out in the same manner as in Example 4, except that Allyl bromide was used instead of Methallyl bromide to obtain Compound 5.

_{^{1 H-NMR (CDCl 3,}} δ): 8.07 (1H, d, J = 7Hz), 7.65 (2H, m), 7.58 (1H, m), 5.27 (1H, m), 3.29 (1H, dd, J = 10, 15 Hz), 2.75 (1H, dd, J = 7, 15 Hz), 1.59 (3H,

Example 6: Synthesis of Compound 6

Sodium peroxide (Na ₂ O ₂ ) (1.95 g, 25 mM) was slowly added to the reaction solution under stirring while dissolving 3-chloropropionyl chloride (5.08 g, 40 mM) in ether (20 ml) , And stirred vigorously for 30 minutes. The reaction solution was heated to 0 캜, and ice (7 g) was added thereto, followed by further stirring for 10 minutes. The organic layer was separated, washed once more with cold water (10 mL) at 0 째 C, and washed again with an aqueous solution of NaHCO ₃ at 0 째 C. The organic layer was separated, dried over MgSO ₄ and concentrated under reduced pressure at 0 ° C or below to prepare 3-chloropropionic peracid.

2-Hydroxy-1,4-naphthoquinone (1.74 g, 10 mM) was dissolved in acetic acid (20 ml), and the 3-chloropropionic peracid prepared above was slowly added at room temperature. The reaction mixture was refluxed with stirring for 2 hours, and then the acetic acid was removed by distillation under reduced pressure. The concentrate was dissolved in CH ₂ Cl ₂ (20 mL) and washed with 5% NaHCO ₃ (20 mL). The water layer was extracted one more time with CH ₂ Cl ₂ (20 mL) and combined with the organic layer extracted earlier. The organic layer was dried over MgSO ₄ and concentrated to give Compound 6 as a mixture with 2- (2-Chloroethyl) -3-hydroxy-1,4-naphthoquinone. This was chromatographed on silica gel to obtain a lapachone derivative (Compound 6) (0.172 g) in a pure state.

_{^{1 H-NMR (CDCl 3,}} δ): 8.07 (1H, d, J = 7.6Hz), 7.56 ~ 7.68 (3H, m), 4.89 (2H, t, J = 9.2Hz), 3.17 (2H, t, J = 9.2 Hz)

Example 7: Synthesis of Compound 7

2-Hydroxy-1,4-naphthoquinone (17.4 g, 0.10 M) was dissolved in DMSO (120 ml) and LiH (0.88 g, 0.11 M) was added slowly. At this time, attention was required because hydrogen was generated. The reaction solution was stirred for 30 minutes while confirming that no more hydrogen was generated. Then, the reaction solution was slowly added with Cinnamyl bromide (3-phenylallyl bromide) (19.7 g, 0.10 M) and LiI (3.35 g, 0.025 M) Respectively. The reaction solution was stirred vigorously for 12 hours while heated to 45 캜. The reaction solution was cooled to 10 ° C or less, and then ice (80 g) was added thereto, followed by addition of water (250 ml), and then concentrated hydrochloric acid (25 ml) was slowly added to maintain the pH of the solution at pH> 1. The reaction mixture was dissolved in CH ₂ Cl ₂ (200 mL) and shaken vigorously. The water layer was wastewater treated and the CH ₂ Cl ₂ layer was treated with 2N NaOH aqueous solution (100 mL 2) to separate the water layer twice. At this time, the remaining CH ₂ Cl ₂ layer was extracted with 2N NaOH aqueous solution and used again in Example 8. When the separated aqueous solutions are combined and then adjusted to acidic pH> 2 using concentrated hydrochloric acid, a solid is formed. This was separated by filtration to obtain a Lapachol derivative. The obtained lapachol derivative was recrystallized using 75% EtOH. The lapachol derivative thus obtained was mixed with sulfuric acid (50 ml), stirred vigorously at room temperature for 10 minutes, and then the reaction was terminated by adding ice (150 g). CH ₂ Cl ₂ (60 mL) was added to the reaction mixture, and the mixture was shaken vigorously. The CH ₂ Cl ₂ layer was separated and washed with 5% NaHCO ₃ . The water layer was extracted one more time with CH ₂ Cl ₂ (30 mL), washed with 5% NaHCO ₃ , and combined with the organic layer extracted earlier. The organic layer was concentrated and then chromatographed on silica gel to give pure compound 7 (2.31 g).

_{^{1 H-NMR (CDCl 3,}} δ): 8.09 (1H, dd, J = 1.2, 7.6Hz), 7.83 (1H, d, J = 7.6Hz), 7.64 (1H, dt, J = 1.2, 7.6Hz) , 7.52 (1H, dt, J = 1.2,7.6 Hz), 7.41 (5H, m), 5.27 (1H, dd, J = (1H, m), 2.08 (1H, m), 0.87 (1H, m)

Example 8: Synthesis of Compound 8

In Example 7, the residue was extracted with 2N NaOH aqueous solution and the remaining CH ₂ Cl ₂ layer was distilled under reduced pressure and concentrated. This was dissolved in xylene (30 ml) and refluxed for 10 hours to induce Claisen rearrangement. The xylene was concentrated by distillation under reduced pressure, and the mixture was vigorously stirred at room temperature for 10 minutes while being mixed with sulfuric acid (15 ml) without further purification, and then the reaction was terminated by adding ice (100 g). CH ₂ Cl ₂ (50 mL) was added to the reaction mixture, and the mixture was shaken vigorously. The CH ₂ Cl ₂ layer was separated and washed with 5% NaHCO ₃ . The aqueous layer was further extracted once more with CH ₂ Cl ₂ (20 mL), washed with 5% NaHCO ₃ , and combined with the organic layer extracted earlier. The organic layer was dried over MgSO ₄ , concentrated, and chromatographed using silica gel to obtain pure compound 8 (1.26 g).

¹ H-NMR (CDCl ₃ ,?):? Dd, J = 1.2, 7.6 Hz), 7.70 (1H, dt, J = (1H, d, J = 9.2 Hz), 7.27 (3H, m), 7.10 (2H, td, J = 1.2, 6.4 Hz), 5.38 Hz), 1.17 (3H, d, J = 6.4 Hz)

Example 9: Synthesis of Compound 9

(1.26 g, 15 mM) and 1,8-diazabicyclo [5.4.0] undec-7-ene (3.4 g, 22 mM) were dissolved in acetonitrile Lt; 0 &gt; C. Trifluoroacetic anhydride (3.2 g, 15 mM) was added slowly while stirring the reaction solution, and stirring was continued at 0 ° C. In another flask, 2-Hydroxy-1,4-naphthoquinone (1.74 g, 10 mM) and cupric chloride (CuCl ₂ ) (135 mg, 1.0 mM) were dissolved in acetonitrile (10 ml) and stirred. The previously purified solution was slowly added to the reaction solution, and the reaction solution was refluxed for 20 hours. The reaction solution was distilled under reduced pressure, and then concentrated. The residue was chromatographed on silica gel to obtain pure compound 9 (0.22 g).

_{^{1 H-NMR (CDCl 3,}} δ): 8.11 (1H, dd, J = 1.2, 7.6Hz), 7.73 (1H, dd, J = 1.2, 7.6Hz), 7.69 (1H, dt, J = 1.2, 7.6 (1H, d, J = 3.2 Hz), 7.60 (1H, dt, J = 1.6, 7.6 Hz), 4.95

Example 10: Synthesis of Compound 10

Compound 9 (0.12 g) was dissolved in MeOH (5 mL) and then 5% palladium (5% Pd / C) (10 mg) was added thereto and stirred vigorously at room temperature for 3 hours. The reaction solution was filtered using silica gel to remove 5% palladium (5% Pd / C) and then distilled under reduced pressure to obtain Compound 10.

_{^{1 H-NMR (CDCl 3,}} δ): 8.05 (1H, td, J = 1.2, 7.6Hz), 7.64 (2H, m), 7.54 (1H, m), 3.48 (3H, s), 1.64 (3H, s), 1.42 (3H, s), 1.29 (3H, s)

Example 11: Synthesis of Compound 11

(1.21 g, 50 mM) and 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (1.14 g, 50 mM) were dissolved in carbon tetrachloride (50 ml) Lt; / RTI &gt; The reaction solution was distilled under reduced pressure, concentrated, and then purified by silica gel chromatography to obtain pure compound 11 (1.18 g).

_{^{1 H-NMR (CDCl 3,}} δ): 8.08 (1H, dd, J = 1.2, 7.6Hz), 7.85 (1H, dd, J = 0.8, 7.6Hz), 7.68 (1H, dt, J = 1.2, 7.6 (1H, d, J = 10.0 Hz), 7.55 (1H, dt, J = 1.2, 7.6 Hz), 6.63

Example 12: Synthesis of Compound 12

2-Methyl-1,3-butadiene (Isoprene) (3.4 g, 50 mM) and paraformaldehyde (3.0 g, 100 mM) were dissolved in 1,4-dioxane 20 ml) were placed in a pressure vessel and heated with stirring at 100 占 폚 for 48 hours. After cooling the reaction vessel to room temperature, the pressure vessel was opened and the contents were filtered. The filtrate was concentrated by distillation under reduced pressure and then chromatographed using silica gel to obtain Compound 12 (238 mg) as a 2-vinyl derivative of β-Lapachone.

_{^{1 H-NMR (CDCl 3,}} δ): 8.07 (1H, dd, J = 1.2, 7.6Hz), 7.88 (1H, dd, J = 0.8, 7.6Hz), 7.66 (1H, dt, J = 1.2, 7.6 D, J = 10.8, 17.2 Hz), 5.18 (1H, d, J = 10.8 Hz), 5.17 (1H, 17.2 Hz), 7.52 (1H, dt, J = 0.8, 7.6 Hz) , 2.62 (1H, m), 2.38 (1H, m), 2.17 (3H, s)

Example 13: Synthesis of Compound 13

2-Hydroxy-1,4-naphthoquinone (1.74 g, 10 mM), 2,4-dimethyl-1,3-pentadiene (4.8 g, 50 mM) and paraformaldehyde ) And refluxed with vigorous stirring for 10 hours. The reaction vessel was cooled to ambient temperature and the contents were then filtered to remove paraformaldehyde. The filtrate was concentrated by distillation under reduced pressure, and then the residue was subjected to chromatography using silica gel to obtain Compound 13 (428 mg) as a? -Lapachone derivative.

_{^{1 H-NMR (CDCl 3,}} δ): 8.06 (1H, dd, J = 1.2, 7.6Hz), 7.83 (1H, dd, J = 0.8, 7.6Hz), 7.65 (1H, dt, J = 1.2, 7.6 (1H, m), 2.04 (1H, m), 1.80 (3H, d, , J = 1.0 Hz), 1.75 (1H, m), 1.72 (1H,

Example 14: Synthesis of Compound 14

2-Hydroxy-1,4-naphthoquinone (5.3 g, 30 mM), 2,6-dimethyl-2,4,6-octatriene (20.4 g, 150 mM), paraformaldehyde 50 ml) and refluxed with vigorous stirring for 10 hours. The reaction vessel was cooled to ambient temperature and the contents were then filtered to remove the paraformaldehyde. The filtrate was concentrated by distillation under reduced pressure and then chromatographed using silica gel to obtain Compound 14 (1.18 g) which is a? -Lapachone derivative.

_{^{1 H-NMR (CDCl 3,}} δ): 8.07 (1H, dd, J = 1.2, 7.6Hz), 7.87 (1H, dd, J = 0.8, 7.6Hz), 7.66 (1H, dt, J = 1.2, 7.6 Dd, J = 11.2, 15.2 Hz), 5.80 (1H, broad d, J = 11.2 Hz), 5.59 (1H, d, J = 15.2 Hz), 2.67 (1H, dd, J = 4.8, 17.2 Hz), 2.10 (1H, dd, J = 6.0, 17.2 Hz), 1.97 3H, bs), 1.63 (3H, s), 1.08 (3H, d, J = 6.8Hz)

Example 15: Synthesis of compound 15

Dissolve 2-Hydroxy-1,4-naphthoquinone (5.3 g, 30 mM), Terpinen (20.4 g, 50 mM) and paraformaldehyde (9.0 g, 300 mM) in 1,4-dioxane (50 ml) . The reaction vessel was cooled to ambient temperature and the contents were then filtered to remove the paraformaldehyde. The filtrate was concentrated by distillation under reduced pressure and then chromatographed using silica gel to obtain Compound 15 (1.12 g) which is a tetracyclic o-quinone derivative.

_{^{1 H-NMR (CDCl 3,}} δ): 8.06 (1H, d, J = 7.6Hz), 7.85 (1H, d, J = 7.6Hz), 7.65 (1H, t, J = 7.6Hz), 7.51 (1H d, J = 7.6 Hz), 5.48 (1H, broad s), 4.60 (1H, broad s), 2.45 J = 16.8 Hz), 2.09 (1H, m), 1.77 (1H, m), 1.57 (1H, m), 1.07 (3H, s), 1.03 d, J = 0.8 Hz), 0.96 (1H, m)

Example 16: Synthesis of Compound 16 and Compound 17

2-Hydroxy-1,4-naphthoquinone (17.4 g, 0.10 M) was dissolved in DMSO (120 ml) and LiH (0.88 g, 0.11 M) was added slowly. At this time, attention is required because hydrogen is generated. The reaction solution was stirred for 30 minutes while confirming that no more hydrogen was generated, and then Crotyl bromide (16.3 g, 0.12 M) and LiI (3.35 g, 0.025 M) were added slowly. The reaction solution was stirred vigorously for 12 hours while heated to 45 캜. The reaction solution was cooled to 10 ° C or less, and then ice (80 g) was added thereto, followed by addition of water (250 ml), and then concentrated hydrochloric acid (25 ml) was slowly added to maintain the pH of the solution at pH> 1. The reaction mixture was dissolved in CH ₂ Cl ₂ (200 mL) and shaken vigorously. The water layer was wastewater treated and the CH ₂ Cl ₂ layer was treated with 2N NaOH aqueous solution (100 mL 2) to separate the water layer twice. At this time, the remaining CH ₂ Cl ₂ layer was extracted with 2N NaOH aqueous solution and used in Example 17. When the separated aqueous solutions are combined and then adjusted to acidic pH> 2 using concentrated hydrochloric acid, a solid is formed. This was separated by filtration to obtain a Lapachol derivative. The obtained lapachol derivative was recrystallized using 75% EtOH. The lapachol derivative thus obtained was mixed with sulfuric acid (50 ml), stirred vigorously at room temperature for 10 minutes, and then the reaction was terminated by adding ice (150 g). CH ₂ Cl ₂ (60 mL) was added to the reaction mixture, and the mixture was shaken vigorously. The CH ₂ Cl ₂ layer was separated and washed with 5% NaHCO ₃ . The water layer was extracted one more time with CH ₂ Cl ₂ (30 mL), washed with 5% NaHCO ₃ , and combined with the organic layer extracted earlier. The organic layer was concentrated and then chromatographed on silica gel to give pure compound 16 (1.78 g) and compound 17 (0.43 g).

¹ H-NMR of Compound _{16 (CDCl 3, δ):} δ8.07 (1H, dd, J = 0.8, 6.8Hz), 7.64 (2H, broad d, J = 3.6Hz), 7.57 (1H, m), (1H, qd, J = 6.0, 8.8 Hz), 3.53 (1H, qd, J = 6.8, 8.8 Hz), 1.54 (3H, d, 6.8 Hz), 1.23

¹ of the compound _{17 H-NMR (CDCl 3,} δ): δ8.06 (1H, d, J = 0.8, 7.2Hz), 7.65 (2H, broad d, J = 3.6Hz), 7.57 (1H, m), (1H, quintet, J = 6.4 Hz), 3.16 (1H, quintet, J = 6.4 Hz), 1.54

Example 17: Synthesis of Compound 18 and Compound 19

In Example 16, the residue was extracted with 2N NaOH aqueous solution and the remaining CH ₂ Cl ₂ layer was distilled under reduced pressure and concentrated. This was dissolved in xylene (30 ml) and refluxed for 10 hours to induce Claisen rearrangement. The xylene was concentrated by distillation under reduced pressure, and the mixture was vigorously stirred at room temperature for 10 minutes while being mixed with sulfuric acid (15 ml) without further purification, and then the reaction was terminated by adding ice (100 g). CH ₂ Cl ₂ (50 mL) was added to the reaction mixture, and the mixture was shaken vigorously. The CH ₂ Cl ₂ layer was separated and washed with 5% NaHCO ₃ . The aqueous layer was further extracted once more with CH ₂ Cl ₂ (20 mL), washed with 5% NaHCO ₃ , and combined with the organic layer extracted earlier. The organic layer was dried over MgSO ₄ , concentrated, and then purified by chromatography on silica gel to obtain pure compound 18 (0.62 g) and compound 19 (0.43 g).

Of compound _{^{18 1 H-NMR (CDCl 3}} , δ): 8.06 (1H, dd, J = 0.8, 7.2Hz), 7.81 (1H, dd, J = 0.8, 7.6Hz), 7.65 (1H, dt, J = M), 2.11 (1H, m), 1.71 (1H, m), 7.51 (1H, dt, J = 0.8, 7.2Hz) 1H, m), 1.54 (3H, d, 6.4 Hz), 1.52 (1H, m)

¹ of the compound _{19 H-NMR (CDCl 3,} δ): 8.08 (1H, d, J = 0.8, 7.2Hz), 7.66 (2H, broad d, J = 4.0Hz), 7.58 (1H, m), 5.08 ( (1H, m), 3.23 (1H, dd, J = 9.6,15.2Hz), 2.80 (1H, dd, J = 7.2,15.2Hz), 1.92 , t, 7.6 Hz)

Example 18: Synthesis of Compound 20

2-Hydroxy-1,4-naphthoquinone (17.4 g, 0.10 M) was dissolved in DMSO (120 ml) and LiH (0.88 g, 0.11 M) was added slowly. At this time, attention is required because hydrogen is generated. The reaction solution was stirred for 30 minutes while confirming that no more hydrogen was generated, and then Geranyl bromide (21.8 g, 0.10 M) and LiI (3.35 g, 0.025 M) were added slowly. The reaction solution was stirred vigorously for 12 hours while heated to 45 캜. The reaction solution was cooled to 10 ° C or less, and then ice (80 g) was added thereto, followed by addition of water (250 ml), and then concentrated hydrochloric acid (25 ml) was slowly added to maintain the pH of the solution at pH> 1. The reaction mixture was dissolved in CH ₂ Cl ₂ (200 mL) and shaken vigorously. The water layer was wastewater treated and the CH ₂ Cl ₂ layer was treated with 2N NaOH aqueous solution (100 mL 2) to separate the water layer twice. When the separated aqueous solutions are combined and then adjusted to acidic pH> 2 using concentrated hydrochloric acid, a solid is formed. 2-Geranyl-3-hydroxy-1,4-naphthoquinone was obtained by filtration and separation. The mixture was stirred vigorously for 10 minutes at room temperature in the state of mixing with sulfuric acid (50 ml) without further purification, and then the reaction was terminated by adding ice (150 g). CH ₂ Cl ₂ (60 mL) was added to the reaction mixture, and the mixture was shaken vigorously. The CH ₂ Cl ₂ layer was separated and washed with 5% NaHCO ₃ . The water layer was extracted one more time with CH ₂ Cl ₂ (30 mL), washed with 5% NaHCO ₃ , and combined with the organic layer extracted earlier. The organic layer was concentrated and then chromatographed on silica gel to give pure compound 20 (3.62 g).

_{^{1 H-NMR (CDCl 3,}} δ): 8.05 (1H, d, J = 7.6Hz), 7.77 (1H, d, J = 7.6Hz), 7.63 (1H, t, J = 7.6Hz), 7.49 (1H (t, J = 7.6 Hz), 2.71 (1H, dd, J = 6.0, 17.2 Hz), 2.19 (1H, dd, J = 12.8, 17.2 Hz), 2.13 , 1.63 (1H, m), 1.53 (1H, m), 1.33 (3H, s), 1.04 0.93 (3 H, s)

Example 19: Synthesis of Compound 21

Compound 21 was obtained in the same manner as in Example 1 except that 6-Chloro-2-hydroxy-1,4-naphthoquinone was used instead of 2-Hydroxy-1,4-naphthoquinone.

_{^{1 H-NMR (CDCl 3,}} δ): 8.02 (1H, d, J = 8Hz), 7.77 (1H, d, J = 2Hz), 7.50 (1H, dd, J = 2, 8Hz), 2.60 (2H, t, J = 7 Hz), 1.87 (2H, t, J = 7 Hz) 1.53 (6H, s)

Example 20: Synthesis of Compound 22

2-Hydroxy-6-methyl-1,4-naphthoquinone was used instead of 2-Hydroxy-1,4-naphthoquinone in the same manner as in Example 1 to obtain Compound 22.

_{^{1 H-NMR (CDCl 3,}} δ): 7.98 (1H, d, J = 8Hz), 7.61 (1H, d, J = 2Hz), 7.31 (1H, dd, J = 2, 8Hz), 2.58 (2H, t, J = 7 Hz), 1.84 (2H, t, J = 7 Hz) 1.48 (6H, s)

Example 21: Synthesis of Compound 23

Compound 23 was obtained in the same manner as in Example 1 except that 6,7-dimethoxy-2-hydroxy-1,4-naphthoquinone was used instead of 2-Hydroxy-1,4-naphthoquinone.

_{^{1 H-NMR (CDCl 3,}} δ): 7.56 (1H, s), 7.25 (1H, s), 3.98 (6H, s), 2.53 (2H, t, J = 7Hz), 1.83 (2H, t, J = 7 Hz) 1.48 (6 H, s)

Example 22: Synthesis of Compound 24

Compound 24 was obtained in the same manner as in Example 1 except that 1-Bromo-3-methyl-2-pentene was used instead of 1-Bromo-3-methyl-2-butene.

¹ H-NMR (CDCl ₃ , 隆): 7.30 to 8.15 (4H, m), 2.55 (2H, t, J = 7 Hz), 1.83 1.40 (3H, s), 1.03 (3H, t, J = 7 Hz)

Example 23: Synthesis of Compound 25

Compound 25 was obtained in the same manner as in Example 1 except that 1-Bromo-3-ethyl-2-pentene was used instead of 1-Bromo-3-methyl-2-butene.

¹ H-NMR (CDCl ₃ , 隆): 7.30 to 8.15 (4H, m), 2.53 (2H, t, J = 7Hz), 1.83 0.97 (6H, t, J = 7 Hz)

Example 24: Synthesis of Compound 26

Compound 26 was obtained in the same manner as in Example 1 except that 1-Bromo-3-phenyl-2-butene was used instead of 1-Bromo-3-methyl-2-butene.

¹ H-NMR (CDCl ₃ ,?): 7.15 to 8.15 (9H, m), 1.90 to 2.75 (4H, m), 1.77

Example 25: Synthesis of Compound 27

Compound 27 was obtained in the same manner as in Example 1 except that 2-bromo-ethylidenecyclohexane was used instead of 1-Bromo-3-methyl-2-butene.

¹ H-NMR (CDCl ₃ , 隆): 7.30 to 8.25 (4H, m), 2.59 (2H, t, J = 7Hz)

Example 26: Synthesis of Compound 28

Compound 28 was obtained in the same manner as in Example 1 except that 2-bromo-ethylidenecyclopentane was used instead of 1-Bromo-3-methyl-2-butene.

¹ H-NMR (CDCl ₃ ,?): 7.28-8.20 (4H, m), 2.59 (2H, t, J = 7Hz), 1.40-2.20

Example 27: Synthesis of Compound 29

The compound 5 (8.58 g, 20 mM) synthesized in Example 5 was dissolved in carbon tetrachloride (1000 ml), 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (11.4 g, 50 mM) Lt; / RTI &gt; The reaction solution was distilled under reduced pressure and then concentrated. The red solid was recrystallized using isopropanol to obtain pure Compound 29 (7.18 g).

_{^{1 H-NMR (CDCl 3,}} δ): 8.05 (1H, dd, J = 1.2, 7.6Hz), 7.66 (1H, dd, J = 1.2, 7.6Hz), 7.62 (1H, dt, J = 1.2, 7.6 D, J = 1.2 Hz), 7.42 (1H, dt, J = 1.2, 7.6 Hz), 6.45

Example 28: Synthesis of Compound 30

{J. Org. 4,5-Dihydro-3-methylbenzo [1,2- b ] thiophene was synthesized using p-Benzoquinone and 1- (N-morpholine) propene according to the synthetic method presented in J. Chem., 55 (1990) 4995-5008. furan-4,5-dione {Benzofuran-4,5-dione}. Benzofuran-4,5-dione (1.5 g, 9.3 mM) and 1-Acetoxy-1,3-butadiene (3.15 g, 28.2 mM) thus prepared were dissolved in benzene (200 ml) and refluxed for 12 hours. The reaction solution was cooled to room temperature and then concentrated by distillation under reduced pressure. This was subjected to chromatography using silica gel to obtain pure compound 30 (1.13 g).

_{^{1 H-NMR (CDCl 3,}} δ): 8.05 (1H, dd, J = 1.2, 7.6Hz), 7.68 (1H, dd, J = 1.2, 7.6Hz), 7.64 (1H, td, J = 1.2, 7.6 1.2 Hz), 7.43 (1H, td, J = 1.2,7.6 Hz), 7.26 (1H, q, J = 1.2 Hz)

Example 29: Synthesis of Compound 31 and Compound 32

4-methylbenzo [1,2- b ] furan-4,5-dione {Benzofuran-4,5-dione} (1.5 g, 9.3 mM) of Example 28 and 2-Methyl- 3-butadiene (45 g, 0.6M) was dissolved in benzene (200 mL) and refluxed for 5 hours. The reaction solution was cooled and then concentrated thoroughly by carcass distillation. This was dissolved again in carbon tetrachloride (150 ml), 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (2.3 g, 10 mM) was added and refluxed for 15 hours. The reaction solution was distilled under reduced pressure in the cooled state and concentrated. This was subjected to chromatography using silica gel to obtain pure compound 23 (0.13 g) and compound 24 (0.11 g).

¹ H-NMR of Compound _{31 (CDCl 3, δ):} 7.86 (1H, s), 7.57 (1H, d, J = 8.1Hz), 7.42 (1H, d, J = 8.1Hz), 7.21 (1H, q , J = 1.2 Hz), 2.40 (3H, s), 2.28 (1H, d,

¹ H-NMR of Compound _{32 (CDCl 3, δ):} δ7.96 (1H, d, J = 8.0Hz), 7.48 (1H, s), 7.23 (2H, m), 2.46 (3H, s), 2.28 (1H, d, J = 1.2 Hz)

Experimental Example 1: Preparation of experimental animals (I)

C57BL / 6 rats aged 8 weeks were used in this experiment. All rats were housed in an aseptic animal room at constant temperature (22 ~ 26 ℃) and humidity (55 ~ 60%). In the experimental group, cisplatin was intraperitoneally injected at a concentration of 20 mg / kg mouse weight (mg / kg or less) in the mice injected with phosphate-buffered saline (PBS) (CDDP), cisplatin and Compound 1 were orally administered at 10, 20 and 40 mg / kg (CDDP +? L) and Compound 1 alone were administered orally (? L).

Experimental Example 2: Tissue Damage Change with Cisplatin Treatment Time

Cisplatin was intraperitoneally injected at a dose of 20 mg / kg and H & E staining was performed.

Referring to FIG. 1, no change or damage was observed in the glomeruli and perfusion tissues until day 2 after the cisplatin injection.

However, in the third day tissue, glomerular abnormality was observed, whereas specific damage by the death of cells in the perfusion tissue was observed (see arrows of CDDP-D3)

Experimental Example 3: Protective effect of compound 1 (β-lapachone, βL) on kidney damage

Compound daily cisplatin was administered, and cisplatin 20 mg / kg was taken 3 days after intraperitoneal injection, and kidney tissues were extracted for tissue analysis.

In Fig. 2, the cisplatin alone group showed considerable damage due to the death of the cells constituting the pericyte. On the other hand, almost no damage was observed in the group treated with Compound 1, and normal tissues such as the untreated group were observed in the single group.

Experimental Example 4: Investigation of Structural Changes in Renal Perfusion Tissue

The degree of damage of renal tissue was compared by PAS assay which detects polysaccharide such as glycogen, glycoprotein and glycolipid present in tissues and confirms normal tissue.

3, normal perfusion tissues were observed in the untreated group or the drug alone group, but the tissue treated with cisplatin was found to be destroyed by the death of tissue cells, It is found that the effect is suppressed.

Experimental Example 5: Investigation of the effect of Compound 1 on biochemical target molecules

To investigate the effect of Compound 1 on the changes in various biochemical target molecules induced by cisplatin, creatinine and urea in blood serum were analyzed and compared.

Referring to FIG. 4, the concentration of creatinine and urea was increased during treatment with cisplatin alone, while the decrease in concentration was observed when treated with Compound 1. [

Experimental Example 6: Change of NAD + / NADH ratio by Compound 1 (I)

Compound 1 is known as a substance that increases NAD + / NADH ratio by increasing the activity of NQO1 (NAD (P) H: quinone oxidoreductase 1) enzyme and is useful for various diseases such as hypertension, diabetes, vascular restenosis, It has been shown to be an effective material in the model. In the present invention, an increase in NQO1 enzyme activity by Compound 1 in an animal model of cisplatin-induced kidney damage is shown in FIG.

Referring to FIG. 5, cisplatin decreased the overall NAD + / NADH ratio in renal tissue, and it was confirmed that this change was restored to some extent by Compound 1.

Experimental Example 7: Change of NAD + / NADH ratio by treatment with Compound 1 (II)

NAD + is known to be used as a substrate for various enzymes, and is known to regulate acetylation, one of the posttranslational modifications of various proteins in cells, as a substrate for Sirt proteins associated with aging and longevity. In the present invention, the acetylation of p65, the target protein of the Sirt1 enzyme in the nucleus, and p53, the target protein of the Sirt3 enzyme in mitochondria, is shown in FIG. 6 to FIG.

6 to 9, the expression of Sirt1 and Sirt3 proteins was increased by cisplatin, which is considered as an adaptive response and is one of the responses of cells to overcome the damage by cisplatin. This expression change appears to be due to the effects of various transcription factors that can be activated by cisplatin, and that this increase in expression is completely regulated by Compound 1. [

In addition, the acetylation of p65 and p53, which are target proteins of Sirt1 and Sirt3, respectively, was markedly increased in the cisplatin-treated group, and a significant decrease was observed in the treated group of Compound 1, and the cause of this change was the activation of NQO1 , The NAD + / NADH ratio in the cells was increased and the activity of Sirt1 and Sirt3, which were increased by the increase of the substrate, was considered to be controlled by the action of Sirt1 and Sirt3, and the acetylation of p65 and p53 proteins in the cisplatin- The expression of the enzyme was increased but the substrate was thought to be due to the small amount of NAD +.

Experimental Example 8: Changes in the vicious cycle of inflammatory mediators by treatment with Compound 1 (I)

Serum and urine levels of inflammatory cytokines, tumor necrosis factor-α, IL-1β (interleukim-1β) and IL-6 (interleukim-6) Was analyzed by an ELISA method, and the results are shown in FIG.

Referring to FIG. 10, cisplatin induces an increase in inflammatory cytokines present in serum and urine, and Compound 1 can confirm the effect of inhibiting the increase of such cytokines in a concentration-dependent manner.

Also, referring to FIG. 11, it can be confirmed that the expression of various inflammatory cytokines in the kidney tissues is remarkably inhibited by the compound 1.

Experimental Example 9: Changes in the vicious cycle of inflammatory mediators by treatment with Compound 1 (II)

Cisplatin-induced renal toxicity is known to occur through increased pathways of reactive oxygen species, DNA adduct formation, mitochondrial dysfunction and inflammatory responses. In the present invention, the expression of p65, an NF-kB subunit associated with activation of NF-kB, which is a typical inflammatory response inducing transcription factor, was observed. As shown in Fig. 12, the expression of p65 in kidney was increased by cisplatin It can be confirmed that compound 1 significantly inhibits the reaction.

Experimental Example 10: Changes of vicious cycle of inflammatory mediators by treatment with Compound 1 (III)

As mentioned above, reactive oxygen species (ROS) are well known as one of the mediators by cisplatin toxicity, and the related proteins of mitochondria and NADPH oxidase (NOX) are typical among the mechanism of production of reactive oxygen species . Actually, the amount of reactive oxygen species generated by cisplatin can be analyzed by an active oxygen species-responsive fluorescent substance such as dichlorofluorescein diacetate (DCFH-DA). In the present invention, The expression of protein NOX 1 and NOX 4 protein was analyzed by immunohistochemistry in kidney tissue, and the results are shown in FIG. 13 and FIG.

13 and 14, a significant increase in expression of NOX 1 and NOX 4 protein was observed in various tissues of the kidney, especially in damaged perfusate tissue by cisplatin treatment, and the increase effect was effectively inhibited by Compound 1 .

Experimental Example 11: Variation of vicious cycle of inflammatory mediator by treatment with Compound 1 (IV)

TLRs (toll-like receptors), one of the receptors for pathogen-associated molecular patterns (PAMPs) important in the immune response, are responsible for many damage-associated molecular patterns (DAMPs) Lt; / RTI &gt; receptors.

Therefore, in the present invention, the expression pattern of TLR4 molecule by cisplatin and compound 1 is observed and shown in Fig.

Referring to FIG. 15, it was observed that cisplatin increased TLR4 expression in tissues, and that the change was remarkably inhibited by Compound 1.

EXPERIMENTAL EXAMPLE 12 Immune Cell-Induced Inflammatory Response Immunized by Renal Tissue by Treatment with Compound 1

The renal toxicity induced by cisplatin is induced by various chemokines, monocyte chemoattractant pretein-1 (MCP-1), and intracellular adhesion molecule 1 (ICAM-1) Induced inflammation increases tissue damage. These infiltration of immune cells can be confirmed by expression analysis of various cell-specific factors (CD11b, F4 / 80, etc.) and MCP-1 and ICAM-1 expressed in kidney tissue cells. .

Referring to FIG. 16, it was observed that the expression of MCP-1 protein was significantly increased in the kidney tissues treated with cisplatin as a whole, and that this change was remarkably inhibited by the pretreatment of Compound 1.

Experimental Example 13: Effect of Compound 2 induced by cisplatin on intestinal toxicity

a) Weight change

After intraperitoneal administration of 3.5 mg / kg of cisplatin intraperitoneally for 4 days (4 to 7 days) with oral administration of 5 ml / kg of solvent (1% CMC) daily, the animals were weighed 10 days (3 days after the last administration of cisplatin) And a decrease of 23.9% is shown in FIG. However, the body weight of normal animals was increased by 26.97g compared to the time of starting the test (24.17g), and thus the body weight was 31.8% lower than that of the control group. In contrast, administration of Compound 2 (10-59 mg / kg) for 3 days before the administration of the anticancer drug showed a tendency to slightly increase the body weight of the animal, and this effect was further increased after administration of cisplatin, Compared with 1.8 ~ 3g of body weight.

b) Long term weight

Referring to Table 2, the absolute weight of the kidneys in the long-term weight after 3 days of administration of cisplatin of 3.5 mg / kg for 4 days was slightly lower than that of the normal animals, but the weight was decreased by 31.8% Is increased in the case of the second embodiment. On the other hand, the absolute weight of the liver decreased to 61.0% of normal animals and significantly lowered relative weight even though body weight was greatly reduced. Especially, the absolute weight of the thymus and spleen decreased to 18.5% and 36.1% of the normal animals, respectively. The relative weights of these organs were also about one-fourth of the thymus and about one-half of the spleen, even considering weight loss.

The increase in the relative weight of kidney by cisplatin showed a tendency to be improved by the administration of Compound 2 at all doses (5-50 mg / kg), and the weight of the thymus and spleen was significantly improved by Compound 2.

[Table 2]

c) Histopathological examination

Referring to FIG. 18, histopathological examination of the small intestine revealed no abnormality in villi and line in normal animals. On the other hand, denaturation of the small villi and severe atrophy were observed in the cisplatin alone group. Nevertheless, it was confirmed that the influence on the line was not large. In contrast, in the group administered with Compound 2, denaturation and atrophy of the small intestine villi were improved in a dose-dependent manner. At 5 and 10 mg / kg, the effect was relatively small, but at 25 mg / kg, And nearly recovered to a level close to normal tissue.

Table 3 shows the results of measurement of villus length, line and depth to quantify the degree of damage to the small intestine. As shown in Table 3, the villus length of the normal control group was 604.0 ± 11.40 μm, and the line and depth were 117.4 ± 2.30 μm, which was 5.14 ± 0.22 μm. On the other hand, the line and length in the group treated with cisplatin of 3.5 mg / kg was 115.6 ± 1.34 μm, which was slightly decreased compared to the control group, while the length of the villus was significantly decreased to 306.0 ± 27.02 μm, μm, which is 48.4% less than the control group. Compared with the compound 2, the cisplatin-treated line and depth were restored, and the villi / line and ratio were significantly improved at all doses, especially when the contracted villus was effectively protected. In other words, the ratio of villus / line increased dose-dependently by compound 2, and at 25 mg / kg, it improved to about 80% of that of the normal control group and recovered to the normal level at a dose of 50 mg / kg.

[Table 3]

(* And ^# are significantly different from the control group ( P < 0.05))

Histopathological examination of the liver revealed no specific lesions as shown in Fig. 19 in the normal control group. On the other hand, in the group administered alone with cisplatin, denaturation of hepatocytes was observed in some animals or infiltration of inflammatory cells was confirmed. In contrast, no abnormal findings were observed in the group receiving Compound 2 at all doses (5-50 mg / kg).

In addition, as shown in FIG. 20, in the normal control group, erythroid, myeloid, and lymphocyte were found in the bone marrow as a result of histopathological examination of the bone marrow producing all the blood cells as well as supplying the lymphocytes with the thymus and spleen. The progenitor cells were tightly packed. On the other hand, in the group administered only with cisplatin, a considerable part of the precursor cells disappeared, and the cellularity of the cells decreased, resulting in a poorly formed bone marrow. In the compound 2 administration group, many cell density improvement effects were recognized at a high dose (50 mg / kg).

Experimental Example 14: Effect of Compound 2 on spleen cell number and function

The results of measurement of the number of splenocytes from the spleen in order to analyze the influence of immune cells upon administration of cisplatin and compound 2 are shown in FIG.

Referring to FIG. 21, in normal animals, the mean was 1,104 ± 220 (× 10,000), but in the group treated with cisplatin only, the number of mice was 302 ± 62, which decreased rapidly to 27.4%. The reduction of these splenocytes by cisplatin was shown to be somewhat defensible by the administration of Compound 2, and the dose of 5, 10, 25 and 50 mg / kg was 422 ± 82, 375 ± 75, 476 ± 86 and 677 ± 118 And recovered to 61.3% of the normal control group at a dose of 50 mg / kg

In addition, the proliferation response to ConA was examined in order to confirm a change in the function of the splenic lymphocyte upon administration of cisplatin and Compound 2, and is shown in FIG.

22, the cells treated with hydrogen peroxide (H ₂ O ₂ , 1 nM), which is an oxidative reactive cytotoxic substance, were treated with conA (1 μg / mL) for 55-60% Respectively. On the other hand, normal cells showed 20.3% proliferation by ConA treatment.

In the animals treated with only solvent CMC (1%) for 10 days, proliferative responses similar to those of normal animals were observed. In the group treated with cisplatin alone, however, the proliferation was lower than that of normal cells not treated with ConA, Lt; RTI ID = 0.0 &gt; T-cell &lt; / RTI &gt; It was observed that the administration of Compound 2 (25 mg / kg) to the cisplatin toxicity was restored to normal.

On the other hand, the proliferative response to LPS was examined in order to confirm a change in the function of splenic B lymphocytes upon administration of cisplatin and Compound 2, and it is shown in FIG. Referring to FIG. 23, the cells treated with hydrogen peroxide (H ₂ O ₂ , 1 nM) showed 26.3% proliferation by LPS treatment.

In the animals treated with only 10 days of solvent, a high proliferative response was observed as in normal animals, whereas a proliferation reaction was not observed in the group treated with cisplatin, indicating a decrease in B cell function by cisplatin. It was observed that this toxicity of cisplatin was restored to normal in the administration of Compound 2 (5 and 25 mg / kg).

Experimental Example 15: Vomiting response of compound 2 according to cisplatin dose

a) Preparation of experimental animals (II)

Marshall Co. (North Rose, USA) for 6 months, followed by about 1 week of laboratory purification, and then weighed 1.5-1.6 kg. Cisplatin alone was subcutaneously injected and compound 2 was orally administered. The cisplatin dose was set to 10 mg / kg by reference. The test substance, Compound 2 (150 mg / kg), was suspended in 1% CMC and orally administered at a dose of 5 ml / kg daily at about 10:00 am for 7 days. Immediately after the last test substance administration, 10 mg / kg of cisplatin was subcutaneously injected subcutaneously to induce nausea and vomiting. CCT was placed at a 60 degree angle to record nausea and vomiting reactions and counted up to 4 hours and 12 hours . And the number of reactions of the test substance to the number of nausea and vomiting reactions when the solvent alone was administered. The time was also measured until the animal died after administration of cisplatin.

(10 mg / kg) and low dose (5 mg / kg) in order to add the notion of cisplatin-induced intestinal damage protection efficacy to the concept of receptor blocker that focuses only on the gastrointestinal effects. That is, the high dose (150 mg / kg) of Compound 2 was administered to a high dose of cisplatin, and the effect of prolonging the mortality time of the animal as well as the endocrine effect was measured. In the case of low dose cisplatin, The survival rate and the protective effect of kidney and liver damage were confirmed.

b) Protection effect against high dose (10 mg / kg) cisplatin

24, when ferrets were administered subcutaneously 10 mg / kg of cisplatin, The mean number of nausea and vomiting was 557.4 times by the first 4 hours and 1,248.5 times by 12 hours. In contrast, when 150 mg / kg of Compound 2 was administered orally for 1 week, nausea and vomiting symptoms caused by cisplatin administration were reduced to 438.2 cycles for 4 hours and 673.7 cycles for 12 hours, respectively, to 78.6% and 54.0%, respectively.

On the other hand, all ferrets died after administration of cisplatin at 10 mg / kg, but in the solvent-only group, they died within an average of 15.7 hours. In contrast, in the group administered with 150 mg / kg of Compound 2, the mortality time due to cisplatin attack was significantly delayed to 47.9 hours.

c) Low dose (5 mg / kg) protection against cisplatin

(25 mg / kg), intermediate dose (50 mg / kg) and high dose (100 mg / kg), respectively. Compound 2 (25, 50, or 100 mg / kg) was suspended in 1% CMC and the body weight was determined by orally administering 5 mL / kg daily at about 10:00 am for 7 days. Immediately after the last test substance administration, 5 mg / kg of cisplatin was subcutaneously injected to induce nausea and vomiting, and CCTV was set up and counted up to 4 hours and 24 hours.

The survival rate was also recorded up to 24 hours after administration of cisplatin, and blood biochemical indicators of kidney and liver were analyzed from serum obtained by centrifuging blood. BUN (blood urea nitrogen) and creatinine were used as indicators of renal toxicity, and AST, ALT, LDH, ALP and TB were measured using hematology analyzer (Hitachi 7080, Japan) as indicators of hepatotoxicity.

25, oral administration of 5 mg / kg of solvent (1% CMC) to normal ferrets for 7 days did not show significant clinical symptoms and showed normal weight gain. After subcutaneous administration of 5 mg / kg of cisplatin after 7 days of solvent administration, the body weight of the animals decreased rapidly from 1.73 kg to 1.36 kg by about 21.8% for 24 hours. It was confirmed that only 10.9% and 7.0% of Compound 2 at 50 mg / kg and 100 mg / kg were reduced, respectively, to such a weight loss, and showed a considerable defense effect.

Referring to Figures 26 and 27, when subcutaneous administration of 5 mg / kg of cisplatin to the ferrets administered only with solvent showed sustained nausea and vomiting, the vomitus was passed until the initial 4 hours There were many cases, but after that, the number of nausea without soil was much higher. As a result of observing these nausea and vomiting symptoms through CCTV, the average response was 190.2 times during the first 4 hours and 609.3 times during the 24 hours. In contrast, when 25, 50 and 100 mg / kg of Compound 2 were administered orally for 1 week, nausea and vomiting symptoms due to cisplatin administration were decreased dose-dependently by 166.4, 156.0 and 110.0 times for 4 hours, respectively, % And 57.8%, respectively. In particular, the number of nausea and vomiting until 24 hours decreased by 469.2, 379.4 and 309.1 in each dose, which was lowered to 77.0%, 62.3% and 50.7%, respectively, compared with the control group.

Referring to Table 4, two animals (40%) out of 5 died after 24 hours from the cisplatin (5 mh / kg) challenge. In contrast, 20 mg / kg of Compound 2 alone resulted in 20% survival, particularly above 50 mg / kg, with a 100% survival rate.

[Table 4]

As shown in Table 5, BUN (9.2-fold) and creatinine (6.0-fold), which are indicators of renal toxicity, increase more than normal values at 24 hours after only 7 days of solvent administration and cisplatin (5 mg / kg) Severe renal toxicity was estimated. In contrast, BUN and creatinine were significantly decreased in the group administered with 25-50 mg / kg of Compound 2, and recovered to a normal level in the group treated with 100 mg / kg.

In the liver injury index, aspartate transaminase (ALT), alanine transaminase (ALT) and lactate dehydrogenase (LDH) rapidly increased to 14.2 times, 12.6 times, and 1.7 times, respectively, in normal animals after administration of cisplatin. ALP (alkaline phosphatase) and TB (total bilirubin), which are the indicators of cholestasis, also increased to 5.0 and 8.7 times of the control group, respectively. Administration of 25-50 mg / kg of Compound 2 to these elevated levels significantly alleviated hepatocyte injury and cholestatic indices. At 100 mg / kg high dose, most of the liver damage index was restored to normal level, and excellent liver protection efficacy was confirmed.

[Table 5]

(* And ^# are significantly different from the control group ( P < 0.05))

Experimental Example 16: Inhibitory effect of Compound 1 on gastric acid secretion

a) Preparation of experimental animals (III)

Animals were placed in groups of 8 per group as shown in Table 6 to determine the test group. After 38 hours of asphyxiation, the animals were anesthetized with ether, followed by incision under the xiphoid process and ligation of the pylorus with suture technique (Shay et al., 1945). Compound 2 or a comparative substance, pantoprazole (30 mg / kg), of compound 2, 3-step dose (30, 60 or 90 mg / kg) at a 4-step dose (30, 60, 100 or 200 mg / kg) immediately after pyloric ligation And injected into the duodenum. In the control group, only the same amount (5 mg / kg) of the solvent (40% HP? L-CD) was administered. After the administration, the abdominal incision was closed with a wound stapler and the animal was restored.

After 6 hours, the animals were anesthetized with ether and sacrificed and stomachs were removed. The gastric juice was dissected and the gastric juice was collected by pipette and centrifuged at 3,000 rpm for 10 minutes. The amount of gastric juice and pH were measured, and each 500 μl stool was taken. 10 μl each of Topfel reagent (1% dimethylaminoazobenzene) or 1% phenolphthalein indicator was added and titrated with 0.1N NaOH solution to determine the amount of free hydrochloric acid (free HCl) acidity was measured (Faloni de Andrade et al., 2007; Qshida et al., 2007).

[Table 6]

b) Measurement of gastric juice

Referring to FIG. 28, the amount of gastric juice in normal animals for 6 hours is 4.01 ml on average. The amount of gastric juice secretion was dose-dependently inhibited by the administration of Compound 1, which was 69.1% at 30 mg / kg, 53.9% at 60 mg / kg, 42.9% at 100 mg / kg and 40.9% at 200 mg / %. In contrast, pantoprazole at 30 mg / kg0 decreased to 53.0 in the control group.

c) gastric pH measurement

29 and Table 7, the administration of Compound 1 effectively increased pHfmf of the gastric juice compared to the control of gastric juice secretion, but it was 1.85 in the control group but was 2.66, 3.52, 6.70 and 30.0 mg at 30, 60, 100 and 200 mg / 5.12, and peaked at 100 mg / kg, and slightly lower than 200 mg / kg at 100 mg / kg.

[Table 7]

(* Indicates a significant difference from the control ( P < 0.05))

d) Free hydrochloric acid content and total acidity measurement

Referring to FIGS. 30 and 31, the amount of free hydrochloric acid and total acidity were effectively inhibited in proportion to the increase of the pH by Compound 1, which was decreased to 16.7% and 12.0% at 100 mg / kg, respectively, as compared with the control. In contrast, 30 mg / kg pantoprazole was less effective than Compound 1 at 100 mg / kg by reducing free acid and total acidity to 26.0% and 25% of the control, respectively. The amount of gastric juice secreted, the amount of free hydrochloric acid, and the total acidity,₅₀(median effective dose; 50% effective total dose) Were 72, 46 and 47 mg / kg, respectively, and were more effective in inhibiting gastric acid secretion than gastric juice.

Therefore, compound 1 showed a gastric juice inhibitory effect similar to that of pantoprazole (30 mg / kg), which is well known as a proton-pump inhibitor at 60 mg / kg, and showed better efficacy at 100-200 mg / kg. The gastric acid secretion was also better than that of 100 mg / kg pantoprazole. Thus, the excellent efficacy of Compound 1 at an adequate dose is expected to be effective in improving various gastric ulcers associated with gastric acid secretion.

Experimental Example 17: Evaluation of inhibitory effect of Compound 2 on gastric acid secretion

a) Gastric secretion measurement

The test group as shown in Table 8 was carried out, and the result of inhibiting the gastric acid secretion of Compound 2 is shown in Fig.

Referring to FIG. 32, the amount of gastric fluid in the normal animals for 6 hours was 5.75 mL on average. The amount of gastric juice secretion was significantly inhibited by the administration of Compound 2, which decreased to 83.1% at the 30 mg / kg, 47.3% at the 60 mg / kg and 65.2% at the 90 mg / kg. In contrast, when 30 mg / kg of pantoprazole was administered, it decreased to 53% of the control group.

b) Measurement of gastric pH

33 and 8, the administration of Compound 2 effectively increased the pH of the gastric juice, while it was 1.33 in the control group. However, at 30, 60 and 90 mg / kg, Low. On the other hand, pantoprazole at 30 mg / kg lowered gastric juice secretion to 50.6% of the control group and increased the pH to 6.36, whereby Compound 2 showed higher efficacy than pantoprazole at 60 mg / kg.

c) Determination of free hydrochloric acid content and total acidity

34 and 35, the amount of free hydrochloric acid and the total acidity were effectively inhibited in proportion to the increase of the pH by the compound 2, which was decreased to a maximum of 44.5% and 59.0% at 60 mg / kg, respectively, as compared with the control. Thus, the ED ₅₀ of Compound 2 in gastric juice, free hydrochloric acid and total acidity is 56, 75 and > 90 mg / kg, respectively, which is more effective in inhibiting gastric secretion than gastric acid.

[Table 8]

(* And significant difference from control ( P < 0.05))

EXPERIMENTAL EXAMPLE 18 Effect of Compound 2 on Improvement of Gastric Ulcer by Ethanol

The animals were sacrificed for 48 hours and then administered orally with 5-dose doses (1, 3, 10, 30 or 60 mg / kg) of compound 2 or pantoprazole. The control group received the same flavor (5 mL / kg) of solvent (1% CMC). After 30 minutes, ethanol (3 mL / kg) of the stock solution was orally administered to induce gastric ulcer (Kim et al., 2008)

One hour after the administration of ethanol, the animals were anesthetized with ether, sacrificed and then stomachs were extracted. The extracted stomach was inflated with 1% formalin (10 mL) and fixed in formalin solution for 15 min. The stomach was incised along the tongue, and the contents of the stomach were lightly rubbed with Kim wipes, spread on a cork board with a pin, and then examined for damage to the gastric mucosa under a three-dimensional stereomicroscope. The ulcer index (ulcer index) was calculated by measuring the length (mm) of the gastric ulcer (hemorrhagic lesion) lesion and 1 mm of the hemorrhage lesion in the case of petechia hemorrhage. Based on the dose - specific gastric ulcer index, a 50% effective dose was presented.

Referring to FIG. 36 and Table 9, the gastric mucosa showed a severe bleeding reaction at 1 hour after oral administration of ethanol (3 mL / kg) of the undiluted solution, showing an average gastric ulcer index of 55.2 mm. This alcoholic gastric ulcer was excellently relaxed by the administration of Compound 2, which was 27.3 mm (49.5%) at 1 mg / kg, 32.7 mm (59.2%) at 3 mg / kg, 5.6 mm (10.1% ) Showed a decrease in gastric ulcer index and almost complete improvement at 30 mg / kg.

[Table 9]

(* And significant difference from control ( P < 0.05))

Referring to FIG. 37, pantoprazole showed a relatively weak effect of 51.4-69.4% in the control group up to 10 mg / kg, and 36.4% of the control group at 30 mg / kg, Very low. However, at 60 mg / kg dose, the gastric ulcer index was 6.4 mm (11.6% of the control group). Comparing the dose-response effects of compound 2 and pantoprazole against alcoholic gastric ulcer, Compound 2 showed an ED ₅₀ of 1.2 mg / kg and pantoprazole had an ED ₅₀ of 11.7 mg / kg, indicating that Compound 2 was about 10 times stronger than pantoprazole I could.

Thus, Compound 2 effectively inhibited gastric acid secretion and improved the gastric ulcer caused by ethanol. Therefore, it is expected to be an excellent drug for the improvement of gastric ulcer caused by specific causes such as gastric hyperproliferation and hangover.

Those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.

Claims (14)

(a) a pharmacologically effective amount of a compound of formula 1, a pharmaceutically acceptable salt, prodrug, solvate or isomer thereof, and (b) a pharmaceutically acceptable carrier, diluent or excipient, A pharmaceutical composition for the treatment and prevention of an anti-cancer agent side effect disease and / or a gastrointestinal disorder,

(One)
In this formula,
R ₁ and R ₂ are each independently hydrogen, halogen, alkoxy, hydroxy or lower alkyl of 1 to 6 carbon atoms;
R ₃ , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are each independently selected from the group consisting of hydrogen, hydroxy, alkyl of 1 to 20 carbon atoms, alkene or alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, Or two of these substituents may form a cyclic structure by mutual bonding;
n is 0 or 1, and when n is 0, adjacent carbon atoms thereof form a cyclic structure by a direct bond. The pharmaceutical composition according to claim 1, wherein the compound of formula (1) is selected from the following compounds (2) and (3)

(2)

(3)
Wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ and R ₈ are the same as defined in Formula (1). The pharmaceutical composition according to claim 1, wherein R ₁ and R ₂ are each hydrogen. The method of claim 2 wherein the compound of formula (II) wherein R _1, R ₂ and R ₄ is R ₂ and R ₆ have the following general formula 2b, each hydrogen in a compound of the formula 2a, or R _1, each represent a hydrogen Lt; / RTI &gt;

(2a)

(2b) The pharmaceutical composition according to claim 2, wherein the compound of Formula 3 is a compound of Formula 3a wherein R ₁ , R ₂ , R ₅ , R ₆ , R _7, and R ₈ are each hydrogen.

(3a) [Claim 2] The pharmaceutical composition according to claim 1, wherein the anti-cancer agent side effect disease is a disease selected from the group consisting of kidney injury, intestinal injury and vomiting caused by administration of an anticancer drug. The pharmaceutical composition according to claim 6, wherein the anticancer agent is cisplatin. The pharmaceutical composition according to claim 1, wherein the gastrointestinal disorder is gastric ulcer. 9. The pharmaceutical composition according to claim 8, wherein the gastric ulcer is a gastric ulcer caused by gastric acid secretion and a gastric ulcer induced by ethanol. 1. A method of using a compound of formula (1) as defined in claim 1 for the manufacture of a medicament for the treatment or prevention of an adverse side effect disease and / or gastrointestinal disorder. The pharmaceutical composition for oral administration according to claim 1, wherein the compound of Formula 1 is contained in a crystalline structure The pharmaceutical composition according to claim 1, wherein the compound of Formula 1 is contained in an amorphous crystal structure The pharmaceutical composition according to claim 1, wherein the compound of formula (1) or (2) is formulated in the form of fine particles. 14. The pharmaceutical composition according to claim 13, wherein the formulation is carried out by a mechanical milling method using a jet mill and / or a spray drying method

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