JP2008520656A - A composition for preventing, suppressing and treating lung cancer caused by tobacco, comprising an extract of Chosei Doraj as an active ingredient - Google Patents

Abstract

The present invention relates to a long-life drage extract (CK) for lung cancer caused by 4- (methylnitrosamino) -1- (3-pyridyl) -1-butanone (NNK), which is known as a carcinogen contained in cigarette smoke. The present invention relates to the prevention, inhibition (suppression) and therapeutic uses of
The present invention relates to a proliferating cell nuclear antigen (Proliferating cell nuclear antigen), which is an indicator of tumor cell proliferation in the lung, wherein the long-life Doraj extract (CK) is increased by NNK, a lung cancer inducer contained in tobacco smoke. : PCNA) is suppressed, and it has been confirmed that it has an effect of suppressing the generation of lung tumors caused by NNK, a lung cancer inducer, and this type of long-life Doraj extract (CK) is included as an active ingredient, The present invention relates to the use of long-life Doraj extract that can be used for prevention, suppression and treatment of lung cancer caused by tobacco.
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Description

The present invention relates to the use of long-life Doraj extract (CK), and more specifically, known 4- (methylnitrosamino) -1- (3-pyridyl) -1- as a carcinogen contained in tobacco smoke. The present invention relates to the prevention, inhibition (suppression) and therapeutic use of long-life Doraj extract (CK) against lung cancer caused by butanone (NNK).

  Oyster (Platycodon grandiflorum A.DC) is a perennial herb of Campanulaceae, the main root is about 10-15 cm, the diameter is 1-3 cm, and the upper is an irregular stem trace There are grayish brown or milky white, deep wrinkles in the vertical direction, and there are skin and wrinkles in the horizontal direction. It is hard but non-fibrous, so it is easy to break, smells a little, and the taste is slightly spicy (Yuk Chance et al., Contemporary Biopharmaceutical, Seoul, Student Company, 460-461, 1993).

The main pharmacological component of this species is triterpine saponin (platycodin A, C, D, D ₂ , D ₃ ), which accounts for about 3% of the roots. This pharmacological component has already been tested through animal experiments, including antitussive, expectorant action, central nervous system inhibitory action (sedation, analgesia, antipyretic effect), anti-inflammatory action against acute chronic inflammation, anti-ulcer and gastric secretion inhibition action, dilating blood vessels. It has been found that it has an anticholinergic action that lowers blood pressure, a hypoglycemic action, an action to improve cholesterol metabolism, etc. (Chem. Pharm. Bull., 20, 1952 (1972), Chem. Pharm. Bull., 23, 2965 ( 1975), J. Chem. Soc., Perkin trans I, 661 (1984), Sogoigaku, 3, 1 (1951), J. Pharm. Soc. Kor., 19, 164 (1975)). Steroidal compounds such as α-spinasterol, stigmast-7-enol, α-spinasterol glucoside, and polysaccharides such as inulin and beturin are also pharmacological components of kyoukyo and have been isolated.

  Chosei Doraji is the root of more than 20 years. So far, despite the various pharmacological actions of Kyoko, cultivation over a long period of time has been difficult, but in recent years, a technology that can grow long-life Doraj as roots of Kyoko over 20 years has been developed ( Lee Seung-ho, “Perennial Cultivation Method” (Korean Patent No. 045731), can be mass-produced. This long-life Doraji is superior in pharmacological action and bioactive effect compared to a common Japanese Kyokyo with a life span of 2 to 4 years.

The research on the long-term doraji so far includes experiments on anti-diabetic effects (J. Korean Soc. Food Sci. Nutr., 25, 986 (1996)) and experiments on hyperlipidemia (J. Pharmaceutical Society of Japan 93 ( 1973) 1188-1194) and the like, but the research on the suppression and protection of liver damage is continuously conducted.

  Lung cancer occurs frequently between the ages of 40 and 70, and rarely may occur before the age of 40. The generation ratio of men and women was 7: 1 in the 1960s, but compared to this, the ratio of women has been higher since the 1980s. This has been analyzed to be due to an increase in female smoking rates. The cause is unknown, but after many mechanical and experimental studies, occupational exposure such as smoking, asbestos, radon, chromium, nickel, aromatic hydrocarbons, arsenic compounds, and radiation acts as a risk factor. It became clear that. Among them, smoking is said to be the main factor.

  Among carcinogens contained in cigarette smoke, 4- (methylnitrosamino) -1- (3-pyridyl) -1-butanone (hereinafter referred to as “NNK”) is the most specific substance and has been studied through many animal experiments. It has been shown to cause lung cancer (MKK Shivji Cell., 69, 367-374 (1992)), and NNK is the strongest carcinogen nitrosamine contained in tobacco. From these facts, it can be seen that NNK is the biggest cause of lung cancer for smokers.

Proliferating cell nuclear antigen (hereinafter referred to as “PCNA”) whose expression is increased by NNK is an index of tumor cell proliferation occurring at the carcinogenesis stage, and is used as a cofactor for DNA polymerase (DNAase) Involved (H. Kohno et al. Cancer Letters 174., 141-150 (2001)). PCNA increases in cells due to the recovery of damaged DNA at various stages of carcinogenesis and DNA replication for cancer cell growth.

The present inventors effectively prevent and suppress the occurrence of lung cancer by NNK, which is a lung cancer inducer contained in tobacco smoke, recover the lung, protect the lung, and treat and prevent non-toxic lung diseases. As a result of repeated research on pharmaceutical preparations, it has been found that the extract of Nagasei Doraji has a preventive and inhibitory effect on the occurrence of lung cancer induced by NNK, and has completed the present invention. Accordingly, the present invention provides a pharmaceutical composition comprising as an active ingredient a long-life Dorazi extract, which is a safe and non-toxic natural product, that has a preventive and suppressive effect on lung cancer caused by smoking or inhalation of tobacco smoke. The purpose is to provide.

  The present invention will be described in detail below.

  The long-life Doraj extract according to the present invention is characterized by having uses for preventing, suppressing and treating lung cancer caused by tobacco. That is, the long-life Doraj extract according to the present invention can be used for the purpose of effectively preventing and suppressing the occurrence of lung cancer by NNK, a lung cancer inducer contained in tobacco smoke, and recovering it to protect the lung. Features.

  The long-life Doraj extract according to the present invention is prepared by a method well known in the art. As long-lived doraj extract, usually hot water extract or organic solvent extract can be mentioned. Such long-lived doraj extract can be prepared either alone or with the addition of an acceptable herbal material, where pharmaceutically acceptable Can be prepared.

  In general, as long-life doraji powder, after drying the long-life doraji so that the water content is 5% or less and pulverizing it, a powder having a particle size of 0.6 mm or less is selected and used. As the hot water extract, the filtrate obtained by adding about 5 to 10 times water to the long-grown Doraji powder and extracting twice at 90 to 100 ° C for 4 to 6 hours is used as it is or acceptable. A material to which a crude drug material or a crude drug material extract is selectively added is used. As the organic solvent extract, a solution obtained by adding about 3 times as much organic solvent to Nagasei Doraji powder and extracting the filtrate obtained by extraction twice at room temperature under reduced pressure is used. As the organic solvent, ethanol is preferably used.

  The long-lived Doraj extract according to the present invention can produce a pharmaceutical dosage form by a conventional method. In producing the dosage form, it is desirable to mix or dilute the long-lived Doradi extract, which is the active ingredient, with the carrier, or enclose it in a container-shaped carrier. Carriers used as diluents include solid, semi-solid, or liquid substances that act as vehicles, excipients, or medium for the active ingredient. For this reason, dosage forms are refined, pills, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, sterile injections And forms such as sterile powders. Examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxy Examples include benzoate, talc, magnesium stearate, and mineral oil. The dosage form can further include fillers, anti-agglomerating agents, lubricants, wetting agents, perfumes, emulsifiers, preservatives and the like. The pharmaceutical compositions according to the invention can be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.

The pharmaceutical composition according to the present invention can be administered via multiple routes including oral, transdermal, subcutaneous, intravenous or muscle. In the case of humans, the daily dose is usually 10 to 1,000 mg / kg body weight, preferably 10 to 100 mg / kg body weight, and can be administered once or divided into several times. However, the actual dosage will need to be determined based on many related factors such as the route of administration, patient age, gender and weight, health status and severity of the disease.

  Hereinafter, the present invention will be described more specifically based on examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited to these examples.

[Example 1] Preparation of long-life doraji hot water extract After drying the long-life doraji to a moisture content of 5% by weight or less, the powder was selected so as to have a particle size of 0.6 mm or less. Prepared. Distilled water (2 liters) was added to 0.2 kg of long-life Doraji powder, extracted twice at 90-95 ° C. for 5 hours, filtered, and freeze-dried to obtain 100 g of long-life Doraj extract (CK).

[Experimental Example 1] Experimental animal and experimental method The following reagents were purchased and used.
Antibodies against PCNA are from Cell signaling, Tris-base and Bovine serum albumin (BSA) are from USB, Twin 20 (Tween-20) and N, N, N ', N'- Tetramethylethylenediamine (N, N, N ', N'-tetramethyl-ethylenediamine: TEMED) is from Sigma, PVDF membrane (PVDF membrane) is from Pall corporation, xylene is from Duksan, Methyl alcohol Ethyl alcohol was purchased from Hayman and NNK was purchased from Toronto Research Chemicals.

2. As an animal experimental animal, a 6-25 week old 20-25 g experimental mouse (A / J mouse) was purchased from Samtako Biokorea Co., Korea and used freely with feed (purina korea) and water. The temperature of the breeding ground was maintained at 21-24 ° C and the relative humidity was maintained at 40-80%. In addition, the light in the farm was adjusted so that day and night were repeated every 12 hours.

3. Induction of lung cancer with NNK and treatment with long-life Doraj extract (CK) Experimental animals were treated with normal control (control) in 15 groups per group, NNK injected intraperitoneally (NNK 100 mg / kg), and NNK They were divided into groups (NNK 3 mg / kg + CK 10 mg / kg, NNK 3 mg / kg + CK 100 mg / kg) to which the long-life Doraj extract (CK) was administered after intraperitoneal injection. NNK was intraperitoneally injected with 100 mg / kg at the start of the experiment, and another 100 mg / kg was intraperitoneally injected one week later. Nagasei Doraji Extract (CK) was orally administered at 10 mg / kg / day and 100 mg / kg / day three times a week for 23 weeks.

4. Statistical analysis Statistical significance was tested using the one-way ANOVA with t-test method, and a case where the P value was smaller than 0.05 was determined to be significant.

[Example 2] Changes in body weight and lung weight by lung cancer inducer NNK and long-lived dragi extract (CK) Investigation of changes in body weight and lung weight of experimental animals by lung cancer inducer NNK and long-lived dragi extract (CK) In order to do this, the body weight was measured before the treatment with NNK and Chosei Doraj extract (CK), and the weight was measured again 23 weeks after the treatment with NNK and Chosei Draj extract (CK). In addition, the lung weight was measured to examine changes in lung weight due to NNK and long-life Doraj extract (CK).

  The initial body weight before the NNK treatment and the body weight after 23 weeks of the NNK treatment in the group in which the mouse as an experimental animal was injected with the lung cancer inducer NNK and the group in which the long-life Doraj extract (CK) was administered after the NNK injection The results are shown in Table 1.

<Table 1> Changes in body weight of experimental mice

As is clear from Table 1 above, when comparing the body weight before treatment with NNK and Nagasei Doraj Extract (CK) and the body weight after 23 weeks treatment, the weight of the group injected only with the lung cancer inducer NNK was slightly decreased. did. It was found that the group receiving long-lived Doraj extract (CK) after NNK injection recovered the weight loss seen in the group injected with NNK alone, and hardly changed when compared with the untreated normal group. .

[Example 3] Inhibitory effect of long-life Doraj extract (CK) on PCNA expression, which is an indicator of cancer cell proliferation increased by NNK DNA damage by NNK known as a carcinogen and DNA replication for cancer cell proliferation by this In order to investigate the effect of long-lived drage extract (CK) on the expression of PCNA involved in the test, 30 days after injection of NNK to experimental animal mice and administration of long-lived draj extract (CK), The sample was extracted, buffered, and homogenized using a homogenizer. The supernatant was collected using a centrifuge, and the expression level of PCNA protein contained in it was Western blotted. Measured by (Western blot) method.

  FIG. 1 is a diagram showing the expression level of PCNA protein. From this figure, the expression level of PCNA protein, which is an index of cell proliferation increased by NNK known as a carcinogen, is expressed as long-lived doraji extract (CK). It was confirmed that it decreased significantly by administration of.

[Example 4] Inhibition effect of tumor formation in lung tissue caused by lung cancer inducer NNK by long-life Doraj extract (CK) Injecting lung cancer inducer NNK to lung tissue of experimental animals, The number of tumors formed in the lungs in the group to which the long-life Doraj extract (CK) was administered after NNK injection was measured and compared, and further, using hematoxylin-eosin stain (hematoxylin-eosin stain), Compare the lungs of the untreated normal group, the lungs of the group injected intraperitoneally with NNK, and the lungs of the group administered with the long-lived dragi extract (CK) after the intraperitoneal injection of NNK. We investigated the inhibitory effect of pulmonary tumor formation.

FIG. 2 is a photograph showing changes in the lungs of experimental animals depending on whether or not the administration of the long-life Doraj extract (CK) is administered. From FIG. 2, the lung tumor generated by NNK administers the long-life Draj extract (CK). It was found that it was suppressed. FIG. 3 is a graph showing the results of measuring the number of lung tumors. From this graph, it was confirmed that the long-life Doraj extract (CK) has an effect of suppressing the generation of lung tumors. FIG. 4 is a photograph showing changes in the results of lung tissue by hematoxylin and eosin staining. From FIG. 4, lung tumors caused by NNK were suppressed by administration of long-life Doraj extract (CK). It was confirmed that

  As described above, the composition containing the long-lived Doraj extract according to the present invention exhibits a preventive and inhibitory effect on lung tumor generation caused by NNK, which is a potent lung cancer inducer contained in tobacco smoke. Therefore, it can be effectively used as a preventive and therapeutic agent for lung cancer caused by causes such as smoking and tobacco smoke.

It is a figure which shows the change of the expression level of PCNA protein by Western blot (Western blot) method by the propriety of administration of a long-life Doraj extract (CK). It is a photograph which shows the change of the pulmonary of an experimental animal by the propriety of administration of Chosei Doraj extract (CK). It is a graph which shows the change of the number of the tumors produced | generated to the exterior of the alveoli by the availability of administration of long life Doraj extract (CK). It is a photograph which shows the change of the lung tissue by the hematoxylin-eosin stain method (hematoxylin-eosin stain) by the availability of administration of long-life Doraj extract (CK).

Claims (1)

A composition for preventing, suppressing and treating lung cancer caused by tobacco, comprising an extract of Nagasei Doraji as an active ingredient.

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