MANUFACTURING METHOD OF PINE CHARCOAL AND NOVEL USE THEREOF AS AN ANTICANCER AGENT
TECHNICAL FIELD
The present invention relates to a manufacturing method of pine charcoal and a novel use thereof as an anticancer agent. In detail, the present invention relates to the use of the pine charcoal as a novel anticancer agent, wherein the pine charcoal is prepared with complete removal of tar by being carbonized with steam of 100°C injected into a stainless steal furnace of 850°C and the anticancer activity of the prepared charcoal, which means suppressing breast or liver cancer, is ascertained.
BACKGROUND ART
Cancer occurs through following three steps : initiation, promotion and progression. Normal cells are mutated to abnormal cells by carcinogens, and as the abnormal cells increase in geometric progression, the cell division occurs abnormally, resulting in the cancer tissue. Various anticancer agents for chemotherapy, which have been developed until now, have toxicity to tumor cells, as well as normal cells, especially bone marrow cells or intestinal epithelium which, of body cells, show comparatively rapid growth. Because the toxicity of these anticancer agents leads to several adverse effects such as kidney intoxication, emesis, or neurotoxicity, other alternative therapies are needed.
It is known that by its strong adsorption ability, the charcoal powder for medical use is very effective to eliminate harmful gases, useless proteins plumping human body, carcinogens and various chemicals from human body. Also the charcoal powder, which can be found in modern eating habits, makes in-food toxic substances dissolved and excreted, which have been accumulated in the body by
eating the processed foodstuffs. This charcoal has been used in Korean and other oriental countries' traditional medicine, and we can find the use of this charcoal in the Korean or Chinese medical old books. For examples, in "the Sunghaeyangbang", it is described that a prescription containing the charcoal is applied to pains of muscles or joints; in "the Ch'ungumbang", the charcoal powder is used in sudden choking of gullet; and as a folk remedy, the charcoal powder is used in diarrhea, influenza, and tympanitis (Kyungsun Lee, Study on the use of charcoal in folk remedies, 1987). Moreover, the study on the in vitro strong adsorption of activated charcoal to organic and inorganic compounds have been reported (Janes, W.H. et al. Chinical toxicology. 1975, 8:516-528), and now many studies on the use of charcoal based on its pharmacological action is actively progressing.
Prior patents of the charcoal powder are on the activated charcoal as an antidote (US Patent No. 5,681,564), and on manufacturing method of the activated charcoal (US Patent No. 5,391,534). However, the methods disclosed in said patents include too many compounds, and when administered into human body, the charcoal power works intestinal humor.
Until now any study on anticancer activity of the pine charcoal has not been made, and it has never been reported that the pine charcoal might suppress the transference of solid cancer, especially prevent and suppress the transference of breast carcinoma. Furthermore, usual charcoal in folk remedies or activated charcoal in the market has been used in an edible form, but the former is not suitable to long- period taking because of the residual tar component and the precipitation of the charcoal in human intestine, and the latter has strong inhibition against intestinal humor, for examples, digestive juice, hormones, and blood, causing rapid decrease of the stamina in case of patients or children, so as the results, long-period taking is not acceptable and the intended effect cannot be obtained.
Considering said problems in prior arts, the present inventors have achieved the present invention, wherein the charcoal is prepared by the pine tree's being
carbonized in high temperature to remove the tar component, and is confirmed that the prepared charcoal should have strong anticancer effect and no side-effect. The present invention is intended to provide a therapeutic composition containing pine charcoal of an effective anticancer activity. Additionally, the present invention is intended to provide a manufacturing method of said pine charcoal.
DISCLOSURE OF THE INVENTION
The purpose of this invention was achieved through following manufacturing steps and activity tests of the inventive pine charcoal, comprising: pine trees were finely chopped in order to be evenly carbonized from the shell to interior of the pieces, and heated and carbonized in a stainless steel furnace of 850°C, together with the steam at 100°C injected into the furnace, and finally to the high-temperature-carbonized charcoal; prepared charcoal was tested in anticancer activity, compared to oak charcoal; in vivo anticancer activity test of the inventive charcoal was performed by transferring ascites cells to subcutaneous layer of mouse's groin, administering the pine charcoal powder to the mice for 4 weeks, and weighing the transferred tumor cells; to test its effect as an anticancer agent, the pine charcoal granule, which had been produced from the mixture of the prepared pine charcoal powder, eucalypt, olive oil, and brown rice leaves, was orally administered to a terminal cancer patient; to examine poison-neutralizing effect of the pine charcoal, the pine charcoal powder was administered to rats, 5 min after the rats were treated with various concentration of a herbicide; the pine charcoal was administered to patients with diabetes or chronic renal
failure; and a pine charcoal toothpaste and pine charcoal soap was manufactured via adding the pine charcoal to usual toothpastes and soaps, respectively.
The present invention is described by following detailed description.
The present invention includes following steps, comprising: the step of finely chopping pine trees in size of 2-3 cm, heating/carbonizing the chopped pieces in a stainless steel furnace of 850°C, together with the steam of 100°C injected into the furnace for complete removal of the tar component, and finally refrigerating the prepared pine charcoal with liquid nitrogen; the step of preparing the charcoal granule after mixing lOOg of the charcoal powder with 5%(w/w) of eucalypt, 3%(w/w) of olive oil, and 0.01%(w/w) of brown rice leaves, wherein the charcoal powder may be prepared by pulverizing the prepared charcoal; the step of testing the prepared charcoal in its in vivo anticancer activity with comparing to oak charcoal; the step of measuring the charcoal's effect on inhibiting growth of solid cancers by administering the charcoal powder to the mice, which tumor cells were transferred to; the step of examining the change of cancer cells by X-ray and CT after the charcoal granule may be orally administered to breast or liver cancer patients at the terminal stage; the step of comparing poison-neutralizing effects of the pine charcoal on rats, to which the pine charcoal powder may be administered, 5 min after treating the rats with various concentration of a herbicide;
- the step of examining effects of the inventive pine charcoal on patients with diabetes or chronic renal failure; and the step of manufacturing a pine charcoal toothpaste and pine charcoal soap
via adding the pine charcoal to usual toothpastes and soaps, respectively.
The inventive manufacturing method of the charcoal includes coniferous pine trees, and of all kinds of pine trees, preferably red pine tree of 15-20 years old, which may actively secret anti-oxidizing agent the most. In the present invention, the temperature in the carbonizing furnace may be between 600-1200°C, and preferably at 850°C with carbonizing a shade-drying pine tree.
In the present invention, the carbonized pine charcoal may be pulverized in
100-500 mesh of powder, preferably in 120 mesh. In the present invention, the herbicide used in the test of poison-neutralizing effect is diluted liquid sρark(DONGBANG AGRO Corp.).
The inventive charcoal granule may be effective on irremediable patients with toxic chemicals, such as defoliant patients, patients poisoned with agricultural chemicals, or AIDS patients. In the present invention, the high temperature-carbonized pine charcoal in powder form may be added to a usual toothpaste or soap for various functions, and mixed with conventional carriers, resulting in medicines in form of capsule, tablet or pill.
The present invention is illustrated but in no way limited by the following examples.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 shows a schematic diagram of the manufacturing device for the inventive medical charcoal composition.
Figure 2 is a X-ray picture of a breast cancer patient at the terminal stage, whose right breast has been spoiled by the cancer cells.
Figure 3 is a X-ray picture of the breast cancer patient, whose cancer cells
have been vanished by administering the inventive pine charcoal.
Figure 4 is a CT picture of a hepatoma patient at the terminal stage, who has irregular tumor.
Figure 5 is a CT picture of a hepatoma patient, whose tumor has been vanished by administering the inventive pine charcoal.
EXAMPLES
Example 1
Production of high temperature-carbonized charcoal of pine trees To produce high temperature-carbonized charcoal, as described in Fig. 1, the manufacturing device for the inventive medical charcoal was used. The manufacturing device consists of a stainless steal furnace(20) to contain the chopped pine tree; a heating device(30) at the bottom of said furnace; an inlet(lθ) for injecting steam; a liquid nitrogen tank(50) containing liquid nitrogen for sustaining its freshness by preventing oxidation of the charcoal; and an exit(40) for tar, through which the volatile tar, produced in carbonization of pine trees, may be discharged. Therefore, using the manufacturing device, the present inventors carbonized seasoned pine trees at 850°C and pulverized the carbonized pine charcoal in form of 120 mesh powder, wherein the seasoned pine trees was prepared by. seasoning pine trees in the shade for 15-30 days.
Compared to other charcoal produced at higher or lower temperature than said temperature, the charcoal produced by the above-mentioned method can be administered for long period because the inventive charcoal has no inhibition effect on extracellular secretion of such substances as digestive juice or hormones. Another inventive charcoal powder, which has no inactivating function unlike charcoal powder of other kinds of trees or activated charcoal, did not inhibit growth of the helpful intestinal bacteria, but suppressed intestinal noxious germ.
Example 2
Manufacturing the anticancer composition using the inventive pine charcoal
The charcoal prepared by the method described in Example 1 was pulverized to fine powder, and after mixing the prepared powder with 5%(w/w) of eucalypt, 3%(w/w) of olive oil, and 0.01%(w/w) of brown rice leaves, the granule was prepared using a granule-making device.
Example 3
Examining anticancer effect of the inventive pine charcoal Anticancer activity of the charcoal prepared as described in Example 1 was tested in vivo and in vitro. The toxic activity of the inventive pine charcoal on tumor cell line and normal cell line was examined, and the effect of preventing the solid cancer was tested for the mice induced to have solid cancer.
Experiment 1 : Comparing the inventive pine charcoal with oak charcoal in in vitro anticancer activity
As anticancer activity of the charcoal prepared as described in Example 1 was compared to a conventional oak charcoal, in vitro toxicity direct to tumor cell line was investigated. The control compound was adriamycin, an anthraquinone anticancer agent.
MCF-7, MCF-7/ADR, the breast cancer cells and OVCAR-4's, the uterine cancer cells were cultured in EMEM, DMEM, and RPMI-1640 media containing 7.5% of FBS, referred to ATCC. Each well of a plate was filled with lOOμl of the tumor cells in the concentration of lxl02/μl, and in each well, pine charcoal, oak charcoal, or adriamycin as a control were added in various concentration. After this, the plate was incubated at 37°C in 5% of CO2 for 2 days, and toxic activity for each cell line was presented in ED50, the concentration in which cell growth may be inhibited to extent of 50%, compared to tumor growth of control group. As the
experimental result was showed in Table 1, in the cell-toxicity, 48 hr after administering the samples to each kind of tumor cells, the pine charcoal showed similar activity with adriamycin and 2-5 fold higher activity than the oak charcoal. As disclosed in the experiments for NIH3T3, adriamycin(an anticancer agent) has the cell-toxicity for normal as well as tumor cell line, while the pine charcoal has toxic activity for tumor cells and specifically no toxic activity for normal cells.
Table 1
Toxic activity of the inventive pine charcoal for tumor cell line
Furthermore, even if the oak charcoal in this experiment has the same mesh of its particle with the pine charcoal, the oak charcoal may have high hardness, sharp surface, and low absorption into humor, resulting in its accumulation in the cell wall of intestinal tissue instead of its proper working. Therefore, the accumulation of the charcoal in the cell wall caused by long-period taking of the oak charcoal can lead to damage of the cell tissue, seriously loss of colonic function.
The inventive pine charcoal has low hardness and high absorption into humor. Further, the inventive pine charcoal has a different C-C bond structure as compared with any other kinds of the arbor, i.e. the oak after being carbonized, because it has a large quantity of essential amino acids contained in the stem.
Experiment 2 : Measuring effect of the inventive pine charcoal on solid tumor growth inhibition
The charcoal prepared in Example 1 had been administered to several
groups of mice for 3 weeks, and it was checked that similar amount of the charcoal had been uptaken to each group. The antitumor activity of the pine charcoal was measured by following steps : transferring 7 day-grown ascites cells (sarcoma- 180) to subcutaneous layer of mouse's groin; administering various amount of the pine charcoal powder to each group of mice for 4 weeks; killing the mice by means of luxation of the cervical vertebrae; weighing the isolated tumor cells; and calculating tumor growth inhibition ratio % as following equation. In this equation, A is the average tumor weight of the control group, and B is the average tumor weight of the treated group.
Tumor growth inhibition ratio (%) = ( A - B )/A x 100
Table 2
Antitumor activity of the pine charcoal in mice transferred with sarcoma- 180
As described in Table 1, the cancer weight of the control group was 3.72g, and that of the group, which had been treated with the pine charcoal, was decreased in proportion to treated amount of the charcoal powder. The weight of mouse uptaken with 2%(dietary w/w) of the pine charcoal was 2.37 g, which means 36.2% of inhibition ratio compared to the control group, and the weight of mouse uptaken with 5%(w/w) was 1.22g, meaning 67.2% of inhibition ratio. Consequently, it is
confirmed that the inhibition ratio be in proportion to the uptake amount of the inventive charcoal.
Example 4 Clinical tests on various kinds of tumors using the inventive pine charcoal
Of the patients who had suffered from breast or liver cancer and had less than 1 year of survival time, volunteers approved by themselves or his/her protectors were selected to clinically examine the antitumor activity of the pine charcoal granule prepared by the method in Example 2. The inventive charcoal granule was administered in 3 times/day.
Experiment 1 : Clinical test on a breast cancer patient
The object of this test was a patient, Ahn Soonja(Citizen JJD : 440820- 2047826), who, after treated with chemotherapy 5 times after diagnosis of breast cancer at Kyunghee Medical Center in Aug. 2000, had still suffered from tumor cells, who could hardly eat something, and who had been finally pronounced to have 3 months remaining for her life. As shown in Fig. 2, the right side had been seriously occupied with tumor cell. The present inventor had administered the patient the granule prepared in Example 2 for 3 months, and observed change of the tumor cell using X-ray. As the results, the right side breast cancer had almost vanished as shown in Fig. 3, and blood test was also clean, so a decision of complete recovery was made.
Experiment 2 : Clinical test on a liver cancer patient The object of this test was a patient, Jeon Suok(Citizen ID : 401113-
1347915) who had been pronounced to have liver cancer at the terminal stage at Ewha Womans University Medical Center on Oct. 30, 1999. When leaving hospital, she had large tumor cell as shown in the CT picture of Fig. 4. The present inventor
had administered the patient the granule prepared in Example 2 for 8 months, and observed change of the tumor cell using CT picture. As the results, the tumor cell had completely disappeared as shown in Fig. 5.
Example 5
Examining anti-poison effect of the inventive pine charcoal
Anti-poison effect of the pine charcoal prepared in Example 1 was examined. 3 g of the pine charcoal powder and conventional oak charcoal powder were respectively mixed with 3 ml of saline and administered to rats with 260g of average weight, 5 min after 1 ml, 3 ml, and 7 ml of the diluted liquid herbicide(Spark liquid agent, DONGBANG AGRO Corp.) had been independently injected to them. Each experimental group include 20 members of rats, and the percentage of survival was measured, 1 hr after administration of the charcoal powder. The liquid herbicide as above-mentioned contains glyphosate-ammonium, a surfactant, a solvent as valid composition, and was used in 200-fold dilution.
As the results shown in Table 3, the group to which the charcoal had not been administered was completely killed by 7 ml of herbicide, while rats to which the charcoal had been administered had high survival %.
This anti-poison effect of the charcoal can be explained like the pine charcoal eliminates the activity of the poison from the gastrointestines by sudden absorption of the herbicide. Also we can see that if compared to usual oak charcoal, the inventive pine charcoal shows higher anti-poison effect by 2-2.5 times.
Table 3 Anti-poison effect of the pine charcoal on herbicide-poisoned rats
Example 6
Clinical test on a diabetic
The object of this test was a patient, Ahn Jinho(Citizen ID : 490915- 1408516) who had been announced to have diabetes and chronic renal failure at the Catholic University of Korea, Taejon St. Mary's Hospital in Sep. 16, 1998, who could not do usual activity, and to whom the granule prepared in Example 2 had been administered for 1 year. After the treatment, the patient's weight had changed from 45 kg to 53 kg, the normal weight, and his blood sugar and blood lipid concentration became normal. According to this result, it has been proved that with the inventive pine charcoal, the diabetes can be treated.
Example 7
Manufacture of a soap containing the inventive pine charcoal powder The pine charcoal powder (120 mesh) and a conventional soap composition were blended at the ratio 5:95, and then the mixture was extruded at 95°C to be molded in a soap frame, wherein the conventional soap composition included palm oil (81.5%(w/w)), glycerin (1.2%(w/w)), squalene (0.5%(w/w)), tocopherol (0.5%(w/w)), perfume (1.2%(w/w)), glycerol (1.0%(w/w)), water (13.5%(w/w)), CaCl(0.6%(w/w)), and alkali component (0.05%(w/w)).
When the charcoal content exceeds 15%(w/w) in total soap content, cracking of the soap can be observed in molding step, and for this reason, it is
recommended that the charcoal content should be between 1 and 15%(w/w), and most preferably 5%(w/w).
The carbonized pine charcoal has micropores formed with removal of air by heating and the adsorption ability of the pine charcoal originates from the micropores. Therefore, by manufacture of a soap using this pine charcoal, the efficacy of adsorption of waste material can be acquired, and contrary to other porous materials, this inventive charcoal comprises carbon. So in application to skin, the osmotic phenomenon, which can occur because of pH-change of the skin, does not occur and this product can be applied to all types of skin such as dry, normal, and oily skin.
Example 8
Manufacture of a toothpaste containing the inventive pine charcoal
The pine charcoal tooth pastes was prepared by mixing 10%(w/w) of the pine charcoal (120 mesh) in Example 1 with a conventional toothpaste composition, which contains sorbitol (40%(w/w)), distilled water (30%(w/w)); 2%(w/w) of poly ethylene glycol (MW 950-1,600) as a moisturizer; l%(w/w) of sodium carboxymethylcellulose as a gelling agent; 1.5%(w/w) of sodium laurylsulfate as a organic surfactant; 8.4%(w/w) of calcium carbonate as a inaqueous brightener used in the dental pharmaceutics; 6.0%(w/w) of silica; 0.1%(w/w) of maltose and 1.0%(w/w) of peppermint as a sweetener and spice.
The pine charcoal has an intraoral antimicrobial or anti-inflammatory effect; no irritating action to the oral cells; and further efficacies such as preventing periodontal diseases and treating halitosis and xerostomia. Also, methane gas or ozostomia from the intestines and foul breath caused by the remnants in the mouth can be removed by strong adsorption of the pine charcoal.
INDUSTRIAL APPLICABILITY
As above described in the examples and experiments, the pine charcoal in this invention can be prepared in the product for oral administration; administrating the pine charcoal to the patients having terminal breast or liver cancer in approval of themselves or their protectors, leads to its excellent cancer-therapeutic activity, such as effectively suppressing the proliferation and transition of malignant tumor cells; and the health cleaning necessities can be provided by incorporating the pine charcoal with conventional soap or toothpaste composition. Thus the inventive pine charcoal is very useful to the biomedical and pharmaceutical industries.