KR100412424B1 - Use of garlic extract as therapeutic agents for prostate cancer and bladder cancer - Google Patents

Abstract

The present invention relates to the clinical development and use of garlic extracts containing allicin, diallyl disulfide and diallyl trisulfide as active ingredients, and ingestion of garlic or clinical administration of the active ingredient of garlic is a powerful anti-cancer effect such as allicin. The present invention not only treats prostate cancer and bladder cancer patients and prolongs their survival, but also causes the development of human bladder cancer and prostate cancer. It has a very good effect on suppressing or preventing it.

Description

Use of garlic extract as therapeutic agents for prostate cancer and bladder cancer}

The present invention relates to the use of garlic extract comprising allicin, diallyl disulfide and diallyl trisulfide as active ingredients, and more specifically, allicin, diallyl disulfide and diallyl tri when administered to a patient with prostate cancer or bladder cancer. The present invention relates to a novel use of garlic extract for the prevention and treatment of human prostate cancer and bladder cancer due to the strong anticancer and immune enhancing effects of sulfide and the like.

Prostate cancer is the most common malignancy among US and European adult males and is the second most important cause of male cancer deaths worldwide. In recent years, the average life expectancy of men has increased in Korea, and the number of elderly people has increased, and the number of patients diagnosed with prostate cancer and cancer caused by prostate cancer has been affected by the changes in dietary conditions such as the introduction of the latest diagnostic methods of prostate cancer and the increase in the intake of animal food. As the related direct mortality rate is increasing significantly, it is expected to develop into an important social disease in Korea soon.

However, prostate cancer already has distant metastases to bones and lymph nodes at the time of initial diagnosis, in which case the only treatment to date is hormonal therapy (dual testicular removal or continuous LHRH analogues injection) or unfortunately Most patients become hormonal-refractory patients who resist conventional hormonal therapy after this primary treatment, resulting in a very unfortunate prognosis in which more than half of patients die within about 1 to 1 year and 6 months. Therefore, the development of new therapies that can increase the survival rate and improve the quality of survival of patients with prostate cancer, especially hormonal therapy-refractory metastatic prostate cancer, has been steadily required. However, no effective new therapies have been developed. Furthermore, if drugs or preparations are developed that can effectively prevent the development of these prostate cancers, and if these preparations do not have side effects when taken or consumed, they may be very breakthrough and effectively suppress the incidence of human prostate cancer.

Bladder cancer is a malignant tumor in which long-term exposure to carcinogens emitted from smoking and harmful environment is the most important cause. In Korea, the most frequent occurrence of urogenital cancer in Korean men is the fifth most common male cancer. It is a very important cancer, and the incidence and cancer mortality rate of bladder cancer continues to increase. Bladder cancer is classified into superficial bladder cancer, invasive bladder cancer, and metastatic bladder cancer according to the progress of the cancer. Although most of the superficial bladder cancers are detected early, surgery of transurethral bladder cancer surgery should be performed. Even if all of the tissues are excised, the cancer has a very high cancer recurrence rate of about 50 to 70%. Therefore, many complications such as urethral stenosis, bladder perforation, and severe infection are caused by repeated reoperation. Recently, in order to lower the recurrence rate, the patient may additionally undergo intra- bladder chemotherapy with chemotherapy or immunotherapy after primary pneumocystectomy, but it causes very severe hematuria and bladder urinary inflammatory disease and prevents the ongoing progression of bladder cancer. can not do. Invasive bladder cancer has to be treated as a very large surgical procedure for radical bladder cancer extraction. However, it is necessary to wear an artificial bladder replacement and a continuous urinary bag after surgery.

In addition, once a patient is diagnosed with metastatic bladder cancer, systemic chemotherapy is the only treatment, but this treatment also results in very fatal systemic complications. Despite the aggressive treatment of chemotherapy, the patient's prognosis is very poor. Therefore, the development of a new treatment for bladder cancer is very urgent and important, and if a prophylactic agent is developed that can effectively suppress the occurrence of bladder cancer, it can be developed and used as a very innovative bladder cancer prevention and treatment. In particular, if such prophylactic and therapeutic agents are not expensive to produce, have no side effects, and have an effective prophylactic effect, a very breakthrough prevention and treatment may be possible.

The inventors of the present invention provide a novel prophylactic and therapeutic chemical capable of selectively removing prostate cancer cells and bladder cancer cells associated with prostate cancer and bladder cancer in order to solve the conventional problems in treating and preventing such prostate cancer and bladder cancer. In addition, as a result of research and testing of plant extracts, garlic extract has very effective cancer prevention ability over existing treatment methods and therapeutic agents that have a negligible effect and efficiently necrosis or kill cancer cells of prostate cancer and bladder cancer. It was confirmed that the cancer cell growth is inhibited to complete the present invention.

Accordingly, it is an object of the present invention to provide an allicin-containing garlic extract having a very excellent effect on the prevention and / or treatment of human prostate cancer and bladder cancer.

It is another object of the present invention to provide a composition for preventing and / or treating human prostate cancer and bladder cancer comprising the allicin-containing garlic extract as an active ingredient.

The object of the present invention is to prepare a garlic extract containing an active ingredient of allicin, diallyl disulfide and diallyl trisulfide from garlic and administered to animal models of human prostate cancer and bladder cancer to investigate the anticancer activity, the garlic extract is human prostate Achieved by confirming that there is a very good effect in preventing and / or treating cancer and bladder cancer.

Hereinafter, the configuration of the present invention will be described in detail.

1 is a microscopic histologic picture showing the prostate cancer necrosis with prominent cell proliferation of prostate cancer cells after HE staining after intralesional immunotherapy using the present invention allicin-containing garlic extract in PC-3 human prostate cancer animal model to be.

Figure 2 is a very clear prostate cancer cell apoptosis as a histological picture after TUNEL special immunohistochemical test using Apotaq after the intralesional continuous immunotherapy with garlic extract containing the present invention in PC-3 human prostate cancer animal model It is a picture showing.

Figure 3 TUNEL analysis using Apotaq in tissues 2 weeks after the simultaneous intratumoral immunotherapy using the garlic extract containing allicin containing the present invention in the bladder cancer animal model in the bladder cancer cells are clearly bladder cancer Microscopic histologic picture showing cell proliferation.

4 is a bladder cancer with bladder cancer cell death in the TUNEL-specific immunohistochemical test using Apotaq after continuous intratumoral immunotherapy using the present invention allicin garlic extract to the tissue of the bladder cancer established in the bladder cancer animal model Necrosis of tissue was observed to confirm the effect of bladder cancer.

The present invention relates to a composition for preventing and treating human prostate cancer, which comprises a garlic extract composed of allicin, diallyl disulfide, and diallyl trisulfide separated and purified from garlic. The present invention also relates to a composition for the prevention and treatment of human bladder cancer comprising garlic extract consisting of allicin diallyl disulfide and diallyl trisulfide separated and purified from garlic. Investigating anticancer activity; Comprising the step of investigating the anticancer activity of the garlic extract of human bladder cancer.

The present inventors crushed 500 kg of fresh garlic and mixed it with the same amount of rapeseed oil, soaking it for about 4 to 6 days, and then adding anhydrous sodium sulfate at 1/10 weight of fresh garlic to remove water from the mixture, followed by column chromatography. Or purified by preparative thin layer chromatography method. All experimental procedures were performed aseptically for in vivo experiments. Garlic extract containing the final purified allicin component is a clear yellow liquid with a strong odor and unusual garlic orientation. The specific gravity is 0.906-0.913 and the acidity is less than 1.0. The saponification value is 163-180 or more, the iodine value is 94-106, and the active ingredient content of allicin is 0.1% or more. The bacteriological test of the garlic active ingredient extract used in this biological test is repeated before every biological test.E. coli, Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella are used only in the undetected extract, and heavy metal test results should be 30ppm or less and BHC, In the residual pesticide test for DDT, ALDRIN, DIELRIN, and ENDRIN, only products with 0.01 ppm or less were used for the biopsy. In order to ensure the proper concentration of allicin-containing garlic extracts containing allicin, a sterile diluent, which is generally pharmaceutically acceptable, may be included, and the age, weight, symptoms of the disease, and the like when administered to patients with prostate cancer and bladder cancer The dosage and the timing of administration may vary depending on the degree of health and the like.

Allicin-containing garlic extracts prepared as described above are directly applied to prostate cancer cells and bladder cancer cells by directly injecting an injector such as a syringe to the patient by using a method described previously in the prostate cancer lesion, that is, by using a computer or magnetic resonance imaging apparatus or ultrasound. This results in cell death and necrosis, which is expected to inhibit the proliferation and growth of cancer cells by stimulating and activating the local or systemic immune system, as well as inhibiting allicin's direct cancer cell proliferation and growth. This action causes prostate cancer cells, bladder cancer cells, or cancer cell-related antigens to be in contact with a biological response modifier of allicin, either directly or by intercellular reactions, thereby stimulating biological immune cells and inducing a response. do. In addition, it acts directly on the neonatal cancer vessels, which are the mammary glands that nourish and grow prostate cancer or bladder cancer masses, and destroys cancer vessels and blocks the formation of further cancer vessels, effectively destroying and treating prostate cancer and bladder cancer masses. You can get it.

In addition, in normal people, especially elderly men or black people who are more likely to develop prostate cancer, or adult men who have a family history of prostate cancer in their families, allicin-containing garlic extracts are previously applied to the periphery of the prostate using the above administration method as needed. By injecting, the occurrence of prostate cancer can be blocked and suppressed. This method can be injected and carried out simply and accurately without any side effects such as bleeding and pain in the outpatients, and the immune system that can prevent prostate cancer from reacting with garlic allicin or allicin mixture and cells around the prostate In addition, by directly acting on prostate cells in the course of cytopathological conversion to cancer cells, the conversion to prostate cancer cells is effectively blocked, and the occurrence of prostate cancer is suppressed.

The use of allicin-containing garlic extract for the prevention of cancer can be used as a cancer prevention agent for bladder cancer. Adult men and women who are more likely to develop bladder cancer through bladder urethra, especially long-term smoking or leather or leather products. By injecting a certain amount directly into adult men and women who have been working for a long time in a factory producing bladder can effectively prevent bladder cancer without side effects such as damage to the bladder function, bleeding, pain, infection, etc. This preventive effect is due to the anticancer mechanism described above.

The use of allicin-containing garlic extracts as an effective and novel treatment for prostate and bladder cancer is possible by direct injection into cancer-related blood vessels that nourish prostate and bladder cancer as well as direct intra-lesional injection. In patients with prostate cancer and bladder cancer, carotid angiography is used to confirm the exact size, location and number of cancer-related blood vessels, and by administering the above-mentioned allicin-containing garlic extract directly to cancer lesions using carotid angiography. Treatment that does not cause any damage to the tissue is possible. The administration of garlic extracts containing allicin directly through cancer-related blood vessels not only regulates the local immune system in cancer cells but also strongly induces the activation of systemic anti-cancer immune system, resulting in rapid cell death and necrosis of prostate and bladder cancer cells. By bringing the direct and rapid cell growth inhibitory effect of cancer cells will have a very effective anti-cancer treatment effect.

Garlic is the most commonly added food used in general diets and is inexpensive, and unlike general medicines, garlic does not cause any adverse effects on the human body. The continuous systemic immunity in the human body is continuously administered by oral administration of garlic extracts containing allicin as the main ingredient. It will help to regulate the system and improve its function and effectively inhibit and block the development of bladder and prostate cancer. Synthetic preparation of such preparations for oral administration can be prepared by the same method as for preparing general oral administration drugs, etc., and the combination can be easily prepared using the above method in order to preserve the effective effect of the allicin component. Continuous oral administration of the allicin-containing garlic extract can be used as a very effective preventive agent for the development of prostate and bladder cancers, which is another important subject of the present invention.

The liquid extract of garlic containing allicin used in the biological test for the animal model of the prostate cancer associated with the present invention was used for the biological test through the culture test of common bacteria, fungi, Escherichia coli, Pseudomonas aeruginosa, Salmorella, etc. Only complete allicin-containing garlic extracts were used after heavy metal tests and residual pesticide tests (0 ppm) such as BHC, DDT, Aldrin, dieldrin and Endrin.

Garlic extract containing allicin of the present invention can be prepared by mixing with conventional excipients, such as capsules, pills, tablets.

Hereinafter, the specific configuration and operation effects of the present invention will be described in detail by way of examples, but the following examples are merely for the purpose of illustrating the present invention, and the scope of the present invention is not limited only to these examples.

Example 1 Anticancer Investigation of Human Prostate Cancer

To establish an animal model of human prostate cancer, 1 × 10 ⁶ numbers of PC-3 hormone-refractory human prostate cancer cells cultured in T-medium were injected into bilateral dorsal athymic nude (nu / nu) nude mice, The experimental animals were classified into immunotherapy treatment group and untreated tumor control group, and immunotherapy was performed using garlic extract containing allicin in the tumor transplantation site. Group I (15 rats, 30 prostate tumors) received 5 mg of allicin-containing garlic extract (Allicin 0.03) for 5 weeks at 1 week interval (1 day, 7 days, 14 days, 21 days, 28 days) from day 1 after tumor transplantation. mg) Immunotherapy (cumulative dose garlic extract 25 mg: allicin 0.15 mg) was performed, and experimental group II (8 mice) was injected with physiological saline only as a tumor control group. Tumor size was measured at a weekly interval using a caliper. The tumor incidence, tumor growth pattern, histopathologic examination, and animal survival time according to each allicin immunotherapy method were carefully evaluated. Statistical significance was analyzed.

As shown in Table 1, only 13.3% of all the animals in the experimental group I who received immunotherapy as garlic extract containing active allicin for 5 weeks from day 1 after human prostate cancer tumor transplantation developed prostate cancer. Immunization with garlic extracts containing allicin significantly reduced the incidence of prostate cancer compared to experimental group II, which is a control group without treatment. This difference was significantly significant in statistical analysis using Fisher's exact test (P <0.01). ). In addition, the mean volume size of prostate tumors developed after allicin immunotherapy in group I was significantly decreased compared to the average volume size of prostate tumors in group II (Mann-Whitney U nonparametric test, P < 0.01). As a result, active immunotherapy using garlic extracts containing allicin effectively prevented the development of prostate cancer in vivo, and it is very useful as an effective prevention and / or treatment of prostate cancer by effectively inhibiting cancer growth and proliferation even after cancer. Confirmed.

Effect of Intralesional Immunotherapy with Garlic Extracts Containing Allicin in Animal Model of PC-3 Prostate Cancer, a Hormone Resistant Human Prostate Cancer Cell Experiment group classification Frequency of prostate cancer (Day 21) Prostate cancer volume (size) (Day 35, ㎣, mean ± SD) Ⅰ group 4/30 (13.3%) 190.25 ± 68.89 Group II 8/8 (100.0%) 1209.75 ± 217.10 Group 1: Inject garlic extract with allicin (cumulative dose 25 mg) Group II: Inject physiological saline

As shown in Figures 1 and 2, gross treatment and prevention of human prostate cancer was confirmed in pathological and special immunohistochemical tests performed at the expense of animal models, using garlic extract containing allicin. Intralesional immunotherapy very effectively induced necrosis of prostate cancer tissue with prostate cancer cell death

Example 2: Anticancer Investigation of Human Bladder Cancer

Experimental Example 1

To investigate anticancer activity against human bladder cancer, 1 × 10 MBT-2 cells, a rat bladder-derived epithelial cell carcinoma cell line, cultured in the embryos of 24 syngeneic C3H / He female mice⁵After subcutaneous injection by water, the experimental animals were divided into immunotherapy treatment group and untreated tumor control group, and immunotherapy was performed using garlic extract containing allicin in the tumor transplantation site. Group I (12 animals) received 5 mg (Allicin 0.03 mg) immunotherapy with garlic extract containing allicin for 5 weeks at 1 week interval (1 day, 7 days, 14 days, 21 days, 28 days) from day 1 after tumor transplantation. (Cumulative dose 25 mg garlic extract: 0.15 mg allicin), Experimental group II (12 animals) was injected with physiological saline only as a tumor control. Tumor size was measured at weekly intervals using a caliper, and according to each allicin immunotherapy method, the incidence of tumors, tumor growth patterns, histopathologic examination, and survival time of animals were evaluated and statistical significance Was analyzed.

Experimental results As shown in Table 2, only two of the 12 animals (16.7%) were tested in Group I, which received immunotherapy as an active allicin-containing garlic extract for 1 to 5 weeks after transplantation of human bladder cancer tumors. The bladder cancer developed and immunotherapy with garlic extract containing allicin effectively reduced the incidence of bladder cancer compared to the experimental group II, which is the untreated tumor control group in which all 12 animals (100%) developed bladder cancer. The prophylactic effect of bladder cancer was obvious and this difference was very significant in statistical analysis using Fisher's exact test (P <0.05). In addition, no experimental animals were killed during treatment with allicin-containing garlic extracts and died of treatment-related toxicity up to 5 weeks after treatment.

Prevention of Bladder Cancer by Local Intralesional Immunotherapy with Garlic Extract Containing Allicin in Animal Model of MBT-2 Bladder Cancer Experiment group classification Bladder Cancer Incidence (Day 28) Frequency of death of experimental animals after treatment with garlic extract with allicin (toxicity test) Ⅰ group 2/12 (16.7%) 0/12 (0.0%) Group II 12/12 (100.0%) 0/12 (0.0%) Group 1: Inject garlic extract with allicin (cumulative dose 25 mg) Group II: Inject physiological saline

Experimental Example 2

MBT-2 cells, a rat bladder metastasis epithelial cancer cell line, cultured in the embryos of another C3H / He 24 syngeneic C3H / He female rat, an experimental animal.⁵Bladder cancer size ranged from 55 to 65 mm in all 24 animals at day 7 after aqueduct³When the animals were classified into immunotherapy treatment groups and untreated tumor controls, immunotherapy was performed using garlic extracts containing allicin in the tumor graft site. Group I (12 animals) received 5 mg (Allicin 0.03 mg) immunotherapy with garlic extract containing allicin for 5 weeks at weekly intervals (7 days, 14 days, 21 days, 28 days, 35 days) from day 7 after tumor transplantation. (Cumulative dose 25 mg garlic extract: 0.15 mg allicin), Experiment group II (12 animals) was injected with only physiological saline into the bladder cancer tumor as a tumor control group. Tumor size was measured at intervals of one week using a caliper, and the incidence of tumors, tumor growth patterns, histopathological examination, and survival rate were analyzed according to each allicin immunotherapy method. As shown in Table 3, the size and volume of bladder cancer measured 35 days after transplantation of bladder cells into experimental animals was 1534.8 ± 552.4 mm on average in the allicin immunotherapy group.³The size of bladder cancer in bladder cancer animal model treated with physiological saline only was 5120.6 ± 812.5mm.³As a result, it was observed that the volume of bladder cancer was significantly reduced in the allicin immunotherapy group, and this difference was statistically significant (Mann-Whitney nonparametric test, P <0.05). As a result, it was confirmed that administration of allicin-containing garlic extract induced very effective bladder cancer growth inhibition and therapeutic effect in a biopsy using a bladder cancer animal model. In particular, the animals treated with allicin chemotherapy showed superior survival compared to the untreated animals.

Bladder Cancer Therapeutic Effect of Anticancer Immunotherapy in Bladder Cancer Tumors Using Allicin-containing Garlic Extracts Experiment group classification Bladder Cancer Size (Day 35, Mean ± SD) Survival Rate of Test Animals (Day 42) Ⅰ group 1534.8 ± 552.4 3/12 (25.0%) Group II 5120.6 ± 812.5 1/12 (8.3%) Group 1: Inject garlic extract with allicin (cumulative dose 25 mg) Group II: Inject physiological saline

Intralesional immunotherapy using allicin-containing garlic extract in histopathologic and specific immunohistochemical tests performed after sacrificing bladder cancer animal models was performed with prostate cancer cell death as shown in FIGS. 3 and 4. It was confirmed that necrosis of cancerous tissue was induced.

As described through the above examples, the garlic extract of the present invention comprising allicin, diallyl disulfide and diallyl trisulfide as an active ingredient has a very excellent effect on the prevention and / or treatment of human prostate cancer and bladder cancer. Therefore, the present invention is a very useful invention in the biopharmaceutical industry.

Claims (2)

After mixing rapeseed oil in garlic and immersing for 4-6 days, it is prepared by removing water of garlic and rapeseed oil mixture by separating anhydrous sodium sulfate, separating and purifying by column chromatography or pre thin layer chromatography, specific gravity 0.906 ~. 0.913, acid value 1.0 or less, safflower 163 ~ 180, iodine is 94 ~ 106, characterized in that it contains allicin, diallyl disulfide and diallyl trisulfide, prevention of human prostate cancer with garlic extract as an active ingredient And therapeutic compositions. After mixing rapeseed oil in garlic and immersing for 4-6 days, it is prepared by removing water of garlic and rapeseed oil mixture by separating anhydrous sodium sulfate, separating and purifying by column chromatography or pre thin layer chromatography, specific gravity 0.906 ~. 0.913, acid value 1.0 or less, safflower 163 ~ 180, iodine is 94 ~ 106, characterized in that it contains allicin, diallyl disulfide and diallyl trisulfide, prevention of human bladder cancer with garlic extract as an active ingredient Therapeutic composition.