KR20040110039A - Composition and method for the prevention, suppression and treatment of recurrence in urothelial cancers including bladder cancer using deuterium oxide and its products - Google Patents

Abstract

PURPOSE: Provided are a composition and a method for the prevention, suppression and treatment of recurrence in urothelial cancers including bladder cancer using pure deuterium oxide and its mixture with a solvent and an anticancer drug. Therefore, it removes inconvenience from patients suffering from recurrence in urothelial cancers, reduces costs therefor, and increases the age of patients. CONSTITUTION: The method for delaying and suppressing recurrence in urothelial cancers and treating cancers initially is characterized by exposing the region of urothelial cancers to pure deuterium oxide and its mixture with isotonic solutions containing oils and distilled water, saline solution or glucose solution and with an anticancer drug.

Description

Composition and method for the prevention, suppression and treatment of recurrence in urothelial cancers including bladder cancer using deuterium for relapse prevention, reinhibition or treatment of early cancer of bladder cancer oxide and its products}

After Professor Yurei discovered and reported heavy water (deuterium oxide, D ₂ O, heavy water) in 1932, the separation and production of heavy water became possible from the beginning of the 20th century. Deuterium (D) in heavy water (D ₂ O) is about 11% heavier than pure water (H ₂ O) because it has twice the mass of hydrogen (H), and most of the heavy water is below the sea. There have been many studies on heavy water mainly in the physico-chemical world. In addition, although not many, biological research results have been reported intermittently and until recently. However, there is not much research on the medical side.

Deuterium in heavy water is known to slow down the hydrogen energy metabolism in energy metabolism in cells, which means that cell division slows down. High concentrations of heavy water (D ₂ O) are known to interfere with cell division, affect cell membrane function, reduce the uptake of DNA precursors in animal cells, and inhibit DNA synthesis. In mice treated with oral heavy water (D ₂ O), they resulted in shrinkage of xenografted cancer tissues, oral with heavy water and cytostatic agents. As a result of administration, the survival of animals with xenograft cancer tissues was greatly increased compared to the control group. It is known that the viable concentration of D ₂ O varies depending on the species. Some protozoa are viable in heavy water at 70% concentration, and some algae and certain bacteria ( Bacteria can survive in heavy water at 100% concentration. There are no reports of deep-sea fish, but they have a very increased chance of encountering heavy water, but they are well-maintained.

In addition, high concentration of heavy water (D ₂ O) is known to act favorably in the survival of mice with high blood pressure and gamma irradiation induced by salt and ethanol. Heavy water is known to alter drug metabolism and lower genotoxicity. In addition, in the normal people who drink a small amount of 99.9% of pure heavy water (D ₂ O) I was not aware of any abnormalities in the body, but to D ₂ O for some patients will be injected directly into a vein eopeotdago over the body. Although some scholars have conducted research on cancer, the cancers studied so far are those cancers that possess characteristics that are easy to progress and metastasize to advanced cancers such as pancreatic cancer and stomach cancer.

Urothelial cancer is a malignant change in the urinary tract epithelium, and the cause of carcinogenesis is mostly the same regardless of the location of the urinary tract epithelium (urethra, bladder, ureter and renal pelvis). Known. Several chemicals, such as 2-naphthylamine, 4-diaminobiphenyl benzidine and aniline, and tobacco, coffee, saccharin, cyclamate and phenacetin are known to be closely related to the development of bladder cancer. In particular, smoking increases the incidence of urinary tract cancer, including bladder cancer, and has been sympathized by scholars around the world, including Korea. The duration of smoking is proportional to the frequency of bladder cancer. The latent period in which a carcinogen first transmits a signal to the bladder mucosa and the development of the final bladder cancer is different depending on the carcinogen, and the overall time for the carcinogen to come into contact with the urothelium in the bladder. Total time). In addition, since the carcinogen initiates carcinogenesis (開始, initiate) is different, the time to appear as overt cancer (different) is different.

The organs in which the mucosal layer is covered with the urinary epithelium are pyelone, ureters, bladder and urethra. Double bladder is a representative and largest organ with the entire inner mucosal layer covered with urinary epithelium. The ureter and the pyelone, including the bladder, which are covered with the urinary epithelium, are sites where the mucosal changes of these organs can be directly confirmed by an endoscope inserted externally of the body. The mucous membrane of the bladder, the urinary tract epithelium, possesses the same biological characteristics as the urinary tract epithelium of other urinary tracts. The mucosal layer of the normal bladder consists of 6-7 layers of transitional epithelium, some of which are continuously exfoliated into the bladder, and at the base, new transitional epithelium of the bladder is continuously produced. Therefore, whatever stimulus or drug damages the transitional epithelium of the bladder surface, depending on the depth of the injury itself, the damage to the transitional epithelium, limited to the bladder mucosa, is regenerated within a short time and has no damage to the structure and function of the bladder. Does not give. In other words, injury, invasion or death of the transitional epithelium located in the innermost part of the bladder mucosa is not a medical problem.

Representative cancer of urinary tract cancer is bladder cancer. Bladder cancer accounts for about 90% of all urinary tract cancers. The frequency of bladder cancer varies slightly from country to country, but it is the most common urinary tract cancer in Korea, accounting for about 3.5% of human cancers worldwide, and the fifth highest incidence of all cancers in Korea. It is not a rare cancer. The incidence of bladder cancer increases with age, 0.6 per 100,000 people in their 30s and below, but 39 in their 60s, and the frequency increases with age. In Korea, mortality from bladder cancer has been reported to have increased four-fold over the past seven years.

Bladder cancer is most often diagnosed by performing cystoscopy because of characteristic painless gross hematuria and many other symptoms. Recently, however, various biomarkers such as telomerase, hyaluronic acid, cell surface antigen, Lewis X antigen, and DNA ploidy have been developed. The primary treatment for bladder cancer is a complete resection of all the bladder cancers to the superficial muscle layer while visually enlarging the bladder mucosa using a transurethral resectoscope. However, carcinoma in situ, flat superficial cancer, or early cancer, which are difficult to visually diagnose or have no characteristic findings even when enlarged by the cystoscope. ), It is impossible to magnify and observe the lens of the endoscope, and it is almost impossible to perform urethral ablation performed with the naked eye. In order to treat these undetectable bladder cancers before they are converted to overt cancer, adriamycin, mitomycin C, thiotepa, and epirubicin Intravesical injection of several types of anticancer agents such as epirubicin, or infusion of immune enhancing agents such as BCG, interferon, bropirimine, and KLH. Attempts have been made to eliminate or slow the onset of new over cancer.

However, the use of such anti-cancer agents or immune enhancing agents in all bladder tumor patients is not justified considering the side effects of these drugs. In high-risk groups, it is desirable to carry out preventive therapy, but the effect on low-risk groups is known to be low. It is questionable how effective prophylaxis, such as the use of anticancer agents or immunopotentiators, can actually be effective in early or flat superficial cancers, and clinical reports suggest that long-term effects are questionable. It is a state. At the same time, since these drugs are mostly anticancer drugs, many unwanted side effects occur after use, and most of them are unable to receive sufficient treatment.

[Technical Aspects]

Precision-based technology for heavy water purification is already entering the generalization stage, as research has been almost completed by various research teams at home and abroad. However, it is extremely difficult to find examples of medical use of heavy water, but some of them are in the research stage of cancer. The cancers studied so far are those that possess characteristics that make it easy to progress to advanced cancers such as pancreatic cancer and stomach cancer. However, there are no studies on bladder cancer and heavy water, and this study is the first attempt at home and abroad.

The drug that the present inventors intend to develop as a chemotherapy for cancer is targeted to urinary tract cancer, but it is very time consuming to refer to all aspects of urinary tract cancer in technical terms. something to do.

Bladder cancer varies slightly from country to country, but accounts for about 3.5% of human cancers worldwide, and it is the fifth most common cancer among all types of cancer in Korea. The incidence of this cancer is known to increase with age, with 0.6 per 100,000 people in their 30s and 39, and 39 in their 60s, and the frequency of bladder cancer increases with a gradual increase in age. ought. In Korea, mortality from bladder cancer has been reported to have quadrupled in recent seven years. It is known that the incidence of men is about three times higher than that of women, and in aging societies, the ability of older patients to resist the side effects of drugs is reduced. It is an important issue.

It is widely recognized at home and abroad that smoking associated with bladder cancer, which has been tolerated or encouraged in the past for a long time as part of the resale business in Korea, is related to bladder cancer. Many carcinogens in cigarettes need 20 to 30 years to develop into overt cancer after initiating carcinogenesis in normal bladder epithelium. It is known. Currently, there is an active anti-smoking campaign in Korea and abroad, but since this movement has been active recently, the importance of bladder cancer remains unchanged since the incidence of bladder cancer in Korea will continue to be similar to the present for decades. will be.

Once the bladder cancer occurs once the recurrence rate is about 70%, bladder cancer is a major problem at home and abroad due to this high relapse rate. In the past, where the country metaphorically promoted smoking for the purpose of taxation, if there is a possibility that the occurrence of bladder cancer exists now and in the future, if there is a way to prevent the recurrence after the first treatment, It will be of great help to survival for many patients with bladder cancer, both domestically and abroad. The causes of recurrence of bladder cancer include the presence of remnant cancer that has not been treated in primary treatment, the presence of cancer that cannot be identified by endoscopy during surgery, or the transplantation of cancer cells isolated from cancer tissues generated during surgery. (intraorgan transplantation) and the development of new cancers after treatment.

To date, preventing recurrence of urinary tract cancers, including bladder cancers with high recurrence rates, regardless of the cause of the recurrence, is one of the most important tasks. It is important to contact anticancer drugs directly with urinary epithelial cancer or An attempt is made to suppress the recurrence of UTIs by contacting immune stimulants. However, no drug has been found or invented that does not yet have side effects and has an effect of suppressing recurrent urinary tract cancer. Some of the various medical communities around the world are working to develop and invent such relapse inhibitors or prophylactic agents.

The above is a medical condition for preventing recurrence of urinary tract cancer.

Since the kidneys, ureters, and urethra, except the bladder, act as a conduit through which urine passes, the urine excreted in the kidneys has a very short contact time with these organs. It is very unlikely that urine will be reabsorbed into the body in a short time and the amount is minimal even if reabsorbed. However, the bladder is the organ having the largest area among the organs covered by the urinary tract epithelium, and the function of the bladder is to store urine for a long time and then excrete it through the urethra. In other words, the bladder is in contact with urine for a very long time compared with other urinary tract epithelial organs, and thus, the contact time with the carcinogen in the urine is long, and the urinary epithelium is in contact with urine for a long time. Urinary epithelial cancer is the most likely cause. At the same time, if the cross-section of blood vessels distributed in the bladder is exposed to the inner surface of the bladder, the solution in the bladder may be absorbed systemically through the blood vessel for several hours until the blood vessel is closed.

When urinary epithelial cancer develops in organs including bladder cancer described above, 70% -80% of treated patients relapse despite primary treatment such as surgery, and in some cases urinary epithelial cancer progresses to advanced cancer as the recurrence repeats. It also threatens the life of the patient. In bladder cancer, a representative example of urinary tract carcinoma, most of them are superficial cancers limited to the bladder mucosa. 70-80% of superficial rocks recur. The development of therapies and drugs that inhibit or prevent the recurrence of frequently recurring cancers is very important in health care. To this end, various attempts have been made to suppress or prevent recurrence using various anticancer agents or immunopotentiators. However, the side effects that occur when using these methods or drugs are clinically problematic, and the long-term effects of these drugs According to a recent academic report, it is uncertainly evaluated.

In this situation, new drugs and treatment methods for the reinhibition and prevention of urinary tract cancers including bladder cancer, which are the above-mentioned problems, have to be developed, and they have reduced or no side effects compared to existing agents, and have very high cytotoxicity against cancer cells. Higher medications may be the best medicine, and this is the hope of many doctors.

The technical problem to be achieved by the present invention is to develop a drug and treatment method using heavy water to suppress or prevent recurrence of urinary tract cancer and to treat early cancer.

FIG. 1 shows that the typical bladder cancer cell T-24 of urinary tract cancer is transplanted into a culture dish containing 1-2 × 10 ⁵ cells in a normal culture medium, and then, at 24 hours, the normal culture solution is completely removed and the heavy water in the culture medium is After maintaining the concentration of 25%, 50%, 75% and 100%, and incubated for 2 hours, the culture medium containing heavy water was completely removed again, exchanged with normal culture medium, and further cultured for 6 days. As a result, 75% and 100% of bladder cancer cells showed a significant decrease compared to the number of transplanted bladder cancers. In other words, the therapeutic effect is showing high.

FIG. 2 shows that the typical bladder cancer cell T-24 of urinary tract cancer is transplanted into a culture plate containing 1-2 × 10 ⁵ cells in a normal culture medium, and then, the normal culture solution is completely removed, and Maintain the concentrations of 25%, 50%, 75%, and 100%, and adjust the concentration of the anticancer mitomycin C to the concentration of 20mg / 40ml in the culture medium and incubate for 2 hours as shown in FIG. After that, the culture medium containing heavy water was completely removed, replaced with the normal culture medium, and further cultured for 6 days. As a result, when the anticancer agent was mixed in the heavy water, cancer cell growth was delayed compared to FIG. . In 75% and 100%, as shown in Fig. 1, the number of bladder cancer cells is shown to be significantly reduced compared to when the bladder cancer is transplanted. In other words, the therapeutic effect is showing high.

* Description of Major Symbols in Drawings *

Cont.-1: The average number of bladder cancers counted after incubation with normal culture every 2 days with a change in normal culture until the 7th day of culture.

Cont.-2: Bladder cancer cell T-24 was transplanted with 1-2 X 10 ⁵ cells in a culture plate containing a normal culture medium, and then completely discharged the normal culture solution in 24 hours, and then 20mg / 40ml concentration in the normal culture solution. The cancer cells were cultured by exchanging the anticancer agent-culture solution containing mitomycin C of the culture plate for 2 hours, and then completely removing the anticancer agent-culture solution and exchanged with the normal culture medium. The average number of bladder cancers counted on day 7 of cancer cell transplantation.

25%: Bladder cancer cell T-24 was transplanted with 1-2 X 10 ⁵ cells in a culture dish containing normal culture medium, and the culture medium containing 25% heavy water was completely discharged after 24 hours of normal culture solution. After incubation by replacing the anticancer agent-depleted water solution with the concentration of 20mg / 40ml mitomycin C in the culture dish for 2 hours, the anti-cancer drug-dewatered solution solution was completely removed and exchanged with the normal culture solution for 2 days. The number of cancer cells that were cultured with each medium exchanged, and the average number of cancer cells counted on the 7th day of cancer cell transplantation.

50%: Bladder cancer cell T-24 was transplanted with 1-2 X 10 ⁵ cells in a culture plate containing a normal culture medium, and the normal culture solution was completely discharged in 24 hours, and then in a culture solution containing 50% of heavy water. After incubation by replacing the anticancer agent-deuterium culture solution containing 20mg / 40ml of mitomycin C with the culture dish for 2 hours, the anticancer agent-water culture solution was completely removed and exchanged with the normal culture solution for 2 days. The number of cancer cells that were cultured with each medium exchanged, and the average number of cancer cells counted on the 7th day of cancer cell transplantation.

75%: Bladder cancer cell T-24 was transplanted with 1-2 X 10 ⁵ cells in a culture plate containing normal culture medium, and the normal culture solution was completely discharged in 24 hours, followed by a culture solution containing 75% of heavy water. The anticancer drug-heavy culture solution in which the mitomycin C concentration of 20mg / 40ml was dissolved was incubated by replacing the culture plate for 2 hours, and then the anti-cancer drug-heavy culture solution was completely removed and replaced with the normal culture solution for 2 days. The number of cancer cells that were cultured with each medium exchanged, and the average number of cancer cells counted on the 7th day of cancer cell transplantation.

100%: Bladder cancer cell T-24 was transplanted with 1-2 X 10 ⁵ cells in a culture plate containing a normal culture medium, and the normal culture solution was completely discharged in 24 hours, and then in a culture solution containing 100% of heavy water. The anticancer drug-heavy culture solution in which the mitomycin C concentration of 20mg / 40ml was dissolved was incubated by replacing the culture plate for 2 hours, and then the anti-cancer drug-heavy culture solution was completely removed and replaced with the normal culture solution for 2 days. The number of cancer cells that were cultured with each medium exchanged, and the average number of cancer cells counted on the 7th day of cancer cell transplantation.

The present invention, the composition and use of heavy water (heavy water, deuterium oxide, heavy water) for the treatment and reinhibition and prevention of urinary tract cancer, including bladder cancer to achieve the object to solve the above-described problems, Also includes the creation of treatments using heavy water which have not been attempted.

Organs coated with urinary epithelium where the heavy water (deuterium oxide, heavy water) preparation of the present invention is used are urethra (urethra), bladder (bladder), ureters (ureter) and pelvis (腎盂, renal pelvis). The urine produced by the body is concentrated in the kidneys by reabsorption and excretion normally. This resorbed urine has no toxic effects on the body. However, since the osmotic pressure of pure water (H ₂ O) or pure heavy water (D ₂ O) is lower than the osmotic pressure in the blood, if they are reabsorbed directly into the body, it will cause hemolysis and the like and be harmful to the human body. . To avoid such human toxicity, the heavy water diluted with pure heavy water (about 100%) or sterilized water (H ₂ O) is dissolved in sterile non-toxic salts and the concentration of heavy water or heavy water product is When used in the same manner as osmotic pressure, even if some of the re-absorption is used for the purpose of treating and preventing recurrent urinary carcinoma, these reabsorbed heavy agents will be nontoxic to the body. That is, the present invention, all of the heavy agent used in the composition and use of the heavy agent for the treatment and reinhibition of urinary tract cancer, including bladder cancer should be made the same as the blood osmotic pressure of normal people.

Hereinafter, with reference to the accompanying drawings showing an embodiment of the present invention will be described in more detail the present invention.

In Fig. 1, experimental results using a cell line T-24 of bladder cancer, which is a representative of urinary tract carcinoma when only heavy water was used, are observed. In this experiment, 0% (control, normal culture) after completely discharging the previous normal culture at 24 hours after transplanting 1-2 X 10 ⁵ cells in a culture dish filled with bladder cancer cells. Bladder cancer cells were exposed to heavy water for 2 hours instead of being filled with cultures formulated to contain heavy water at 25%, 50%, 75% and 100% concentrations. After 2 hours, all cultures containing heavy water were sucked out and then filled with normal cultures and cultured for 6 more days. Once every 2 days, the old culture was aspirated and drained and replaced with fresh normal culture. Seven days after cancer cell transplantation, cancer cells were detached from the culture dish and counted under a microscope. As shown in FIG. 1, the control group (0%) shows very high growth. In the 50% concentration group, the number of cells transplanted was much higher than that of the first transplanted cancer cells, but in the 75% and 100% groups, the number of transplanted cells decreased rapidly, so that only 1% or less of the cells compared to the control group (0%) Was alive. This was reduced to less than 1/10 of the number of cancer cells transplanted for the first time. However, this experiment was performed only 7 days after cancer cell transplantation. In other words, maintaining the concentration of heavy water in the culture medium to 50%, slowing the growth of cancer cells, but did not kill cancer cells.

However, when the concentrations of heavy water in the culture were maintained at 75% and 100% concentrations, the cancer cells decreased to less than 1/10 of the amount transplanted into the culture dish. This suggests that high concentrations of heavy water possess cancer cell killing effects. As shown in Figure 1, the higher the concentration of heavy water, the slower the growth of cancer cells, the growth of cancer cells in culture medium containing 75% and 100% concentration of heavy water, not the delay of cancer cells transplanted into the culture plate is greatly increased have.

FIG. 2 is an experimental result of bladder cancer cell line T-24 cultured in a culture medium prepared by dissolving 20 mg of antitomycer mitomycin C in 40 ml of heavy water. The concentration of mitomycin seed used to suppress recurrence in bladder cancer patients varies from 10 mg / 40 ml to 40 mg / 40 ml depending on the user. The concentration used in Figure 2 is an experimental result using the intermediate concentration of the concentration used by various doctors. In the experiment of Figure 2, after the transplantation of 1-2 X 10 ⁵ cells in the culture dish filled with bladder cancer cells normal culture 24 hours after completely discharged the normal culture medium, the concentration of heavy water was Heavy water 0%)-1, control group (0% heavy water and anticancer) -2, 25% (25% heavy water and anticancer), 50% (50% heavy water and anticancer), 75% (75% heavy and anticancer) and 100% After exposure of the cancer cells to the culture medium maintained at the concentration of (100% of the heavy water and the anticancer agent) for 2 hours, the culture medium containing the heavy water and the anticancer agent was completely discharged again, replaced with the normal culture medium, and then replaced with the normal culture medium every 2 days. While culturing for 6 days. When the growth of the cancer cells was observed, relative cell growth was severely delayed in Control-2 compared to Control-1. It was observed that the cell growth was severely delayed in the 25% group compared to the control-1. There was no significant difference in cell growth delay between control group 2 and 25% group (containing 25% heavy water concentration and anticancer drugs), but 25% had more cell growth disorders. At a concentration of 50% (containing 50% heavy water and anticancer agent), the delay in cell growth was significantly reduced compared to control-1 and control-2. Growth of cancer cells in culture medium containing 75% and 100% of heavy water and anticancer agent was not significantly different between the two groups. Almost all of them are dead and can't find cancer cells. Comparing FIG. 2 and FIG. 1, it can be seen that the use of heavy water and anticancer agents resulted in greater cancer cell killing effects than the use of heavy water alone in the preparation of a culture medium for bladder cancer cell culture, and the use of low concentration heavy water. It was severe in high concentrations of heavy water.

Already, as described above, various types of anticancer solutions are injected into the bladder for reinhibition of bladder cancer and microcance treatment. Although the short-term or long-term effects of these anticancer agents are still unclear, the purpose of using these drugs is to lower the high recurrence rate of urinary tract epithelial cancer or to prevent recurrence and to treat early cancer.

As shown in FIG. 1, heavy water alone possesses the effect of delaying the growth of UTIs, but the combination of anticancer and heavy water in FIG. 2 has resulted in a delay in synergistic cancer cell growth compared to the use of anticancer drugs alone. .

Therefore, as described above, heavy water preparations can be used alone or in combination with anticancer drugs to prolong (delay) the period of recurrence of urinary epithelial cancer, which has been previously tried with only anticancer drugs, or to improve the treatment of urinary epithelial cancer. .

Solvents and salts used to formulate this heavy water formulation are harmful to the human body, and anticancer agents can use all anticancer drugs currently available on the market.

The present invention uses heavy water, which has never been used to treat cancer of the urological system, to reduce and prevent very high recurrence rate of around 80%, and to treat early cancer, in combination with various solvents including heavy water alone or anticancer agents. It aims at manufacturing the used heavy water preparation.

The present invention, the effect of the composition and use of heavy water (deuterium oxide) preparations for the treatment and reinhibition of urinary tract cancer, including bladder cancer, the recurrence rate, relapse delay and early cancer of urinary tract cancer with a high recurrence rate of about 80% It can be used to treat patients suffering from urinary epithelial cancer, which can eliminate the discomfort associated with recurrence of urinary epithelial cancer, prevent costly expenses, prolong patient's life and improve quality of life. have.

Claims (5)

To delay the recurrence of urinary tract cancer, including bladder cancer, to suppress recurrence and to treat early cancer, Treatment method in which heavy water (deuterium oxide) or heavy water preparation is exposed to urinary epithelial cancer site for a certain time The method of claim 1, Method of contacting urinary epithelial carcinoma with heavy water (deuterium oxide) alone to inhibit, delay or prevent recurrence of urinary carcinoma, and to treat early cancer The method of claim 1 Mixed preparations used in combination with heavy water (deuterium oxide) to form a mixture include various types of isotonic solutions, including salts, sterile distilled water, physiological saline or glucose solutions, and normal cells. Other solutions that do not damage the skin, such as exposure to urinary epithelial cancer to inhibit the recurrence, delay or prevent the recurrence of urinary carcinoma, including bladder cancer, used to treat early cancer. The method of claim 1 Combination preparations used in combination with heavy water (deuterium oxide) to form a mixture include various isotonic solutions, including vegetable, animal or mineral oils, and normal cells. Other intact solutions, including those that are harmless to the human body, such as sesame oil, cottonseed oil, peanut oil, palm oil, coconut oil, and the like, are diluted in heavy water (deuterium oxide) and exposed to urinary epithelial cancer. Used to inhibit relapse, delay or prevent recurrence, or treat early cancer of urinary tract cancer, including bladder cancer The method of claim 1 It is a mixed preparation used in combination with heavy water (deuterium oxide) to form a mixture. It is a conventional anticancer agent that is dissolved in heavy water (deuterium oxide) and exposed to urinary tract epithelial cancer. Used to inhibit, delay or prevent relapse, or treat early cancer.