The anticancer drug and the method of its production

 

(57) Abstract:

The invention relates to medicine, in particular to cancer pharmacology. The invention consists in the fact that developed a new drug on the basis of native squalene, derived from the fat of deep-sea sharks or olive oil. Native squalene activate with getting active derivatives with antitumor activity. Activation is carried out using heat, UV or IR radiation exposure to reactive oxygen species. The technical result of the present invention is to create a new anticancer drug based on activated squalene, do not have a cytotoxic action on healthy cells. 2 S. and 2 C.p. f-crystals, 8 tab., 13 Il.

The invention relates to medicine, namely to clinical pharmacology, in particular to the development of a new drug for cancer treatment based on activated squalene and how to obtain it.

The incidence of malignant tumors in recent years has increased in Russia by more than 10%. Every year there are about 500 thousand sick again. The mortality rate of cancer of actualnode remission, reducing complications and expansion of the range of anticancer agents.

Pharmacotherapy of tumor pathology, along with radiation therapy and surgery, is the most important component of the fight against cancer. In recent years it has been enriched with many new drugs that enhance its effectiveness. However, one of the limiting points in the medicinal treatment of malignant tumors is adaptation of tumor cells to drugs.

All antineoplastic (or protivoblastomna) drugs are divided into several groups based on their chemical structure, mechanism of action, sources: alkylating agents, antimetabolites, antibiotics, agonists and antagonists hormones, alkaloids and other plant.

Recently, much attention has been attracted endogenous antitumor compounds. Discovered the efficacy of interferon in certain types of tumors studied the antitumor activity of other lymphokines.

Along with specific inhibitory effect on the tumor, modern anti-cancer drugs act on other tissues and article to use them in other areas of medicine. In addition, a significant drawback of modern drugs is the low selectivity of action against tumor cells.

One of the main side effects of anticancer chemotherapy is bone marrow suppression, which requires accurate control of the dose and mode of use of drugs; you need to consider that depression of haematopoiesis enhanced by combination therapy, combination drugs with radiation therapy and other frequent nausea, vomiting, loss of appetite, diarrhea, possible alopecia and other side effects. Some antitumor antibiotics have cardio (doxorubicin and others), nephro - and ototoxicity. When using certain drugs may development of hyperuricemia. Estrogens, androgens, their analogues and antagonists can cause hormonal disorders.

One of the characteristic features of many anticancer drugs is their immunosuppressive effect, which can weaken the body's defenses, to facilitate the development of infectious complications. At the same time, a number of anticancer agents (methotrexate, azathioprine, cyclophosphamide, cytarabine, prospidin and others) is used as immunosuppressants in AU Vlada expressed leuko - and thrombocytopenia, severe cachexia, severely impaired liver function and kidney failure, end-stage disease. The question about their use during pregnancy is decided individually. Usually, due to the risk of teratogenicity of these drugs during pregnancy is not appointed, as with breastfeeding (you should stop breastfeeding).

Apply anti-cancer drugs prescription-only medical oncologist. Depending on the nature of the disease and its course, efficacy and tolerability of chemotherapy can change the destination schema, dose, be combined with other drugs, etc.

Recently developed drug methods to improve the effectiveness and tolerability of anticancer drugs. So, calcium folinate allows you to optimize the action of methotrexate and other antimetabolites (including isomers), a new highly effective anti-emetics (blockers of serotonin 5-HT3receptors: ondansetron, tropisetron and others) to reduce nausea and vomiting, "colony-stimulating factors (filgrastim, molgramostim, and others) - to reduce the risk of neutropenia.

For treatment of oncological diseases is wide enough the tsya vinblastine and vincristine the alkaloids isolated from the plant Vinca rosea, kolhamin and colchicine - from corms of Colchicum Speciosum, podophyllum from rhizomes with roots of Podophyllum peltatum. The action of these alkaloids is due to the ability to bind with molecules of tubulin, inhibit the formation of masnago spindle and block mitosis, i.e., to stop the mitotic cell division at metaphase [1].

Antitumor pharmacology and molecular pharmacotherapy of cancer is constantly evolving and achievements in recent years explicitly talk about it. In particular, researchers from Germany (Reszka R. and others) has developed an antitumor agent, representing a cytostatic in liposomal form, optionally containing contrast agent selected from the group of: iodine, gadolinium or magnetite (U.S. Pat. US 6207133 from the 27.03.2001). Very effective in vivo are considered and the so-called "heat shock protein (hsp), in particular, such proteins as hsp60, hsp90, is effective in doses of 10-1000 μg. It is necessary to take into account their individual histocompatibility specific organism (U.S. Pat. US 6136315, 24.10.2000).

The use of these tools due to their properties: cytotoxicity to normal cells, rejection of foreign proteins can prove difficult when the building anticancer drug based on a natural, physiologically acceptable agent, capable and reasonable treatment, to serve as an effective remedy for various forms of tumors. The basis for the creation of such substances was the fat of deep-sea sharks, known since ancient times.

Fishermen ancient Japan called it "Same dawa" or "panacea". The sharks, which they caught on the bottom of the Bay, the depth of 1000 meters, was delightful meat. They are especially valued for their fat, which was used regularly to improve health. In 1758 the famous botanist and zoologist Carl Linnaeus announced great health value and health value of the extract fat of deep-sea sharks. On the Mediterranean Islands "miracle oil" for centuries it was a remedy for various diseases, and also as a source of strength and energy. In the Scandinavian countries have traditionally used the fat shark fishermen for wound healing, treatment of respiratory diseases and inflammations of the rectum. In China, the extract fat of deep-water shark was described in ancient pharmaceutical book "Honeycake". In Spain the ancient sailors regularly took "ACE ite de bacalao" or "the fat of the great fish" for resistance to colds and other diseases.

In 1906 Dr. Mapumulo Trueim assunnah hydrocarbons. He later called them "Squalene" - squalene.

The study of squalene in the last decade have shown that this highly biologically active agent has a number of useful mammalian therapeutic and cosmetic properties, but at the same time, its use for a man must be very careful, because itself squalene may under certain conditions to have adverse impacts. Below we provide specific examples of studies supporting those and other properties of the squalene.

Squalene is an antiviral compound for the treatment of native hepatitis C virus, it improves in vitro replication of hepatitis C virus (U.S. Pat. US 5858389, 12.01.1999).

The data obtained about the possibility of using squalene as antimutagen. Squalene at a concentration of 80 micromoles almost completely suppresses the clastogenic effect of sodium arsenite.

A number of studies devoted to the use of squalene to reduce the level of cholesterol in the blood [3, 4], as well as U.S. Pat. EP 0521103, 1994.

Squalene has repellent properties [5].

Squalene is widely used as adjuvant in vaccination (U.S. Pat. US 5109026, 1992; US 5733555, 1998; EP 0745388, 1996).

Studied in detail the influences passing through the skin of piroxicam [6].

Very important for our study is the fact that the high content of squalene in natural olive oil is a major factor in the reduction of risk (i.e., prevention) cancer [7].

Most of the described studies conducted in vitro or on animals, as squalene, as mentioned above, in addition to the useful properties and has adverse effects on the human body.

In particular, the data computed tomography showed that in patients with confirmed exogenous lipoid pneumonia caused by the inhalation of squalene (product processing shark liver oil) [8, 9].

Some experiments show that squalene inhibits the induction of neural transmission [10] and causes encephalopathy in rats [11].

The contradictory effects of squalene on the human body is most clearly illustrated by the study of the so-called GWS (Gulf War Syndrome) [12].

During the Gulf war (Iraq, 1991) U.S. servicemen were detected symptoms of an unknown disease: memory loss, thyroid disease, allergies, weakness, rash, persistent pain. It is assumed that the disease is caused by vaccines containing squalene. Postradavshix from syndrome GWS registered a high level of antibodies, due to high content. In the blood of the veterans who had not experienced the syndrome, such antibodies are not detected.

Vaccination may be the cause of the syndrome because soldiers were systematically vaccinations against plague and anthrax, avoiding the Iraqi biological weapons. Vaccinations not only did French soldiers, only the French veterans syndrome GWS is not detected. Researchers from the UK (King's College Hospital, London) note that the above vaccinations can cause syndrome GWS British veterans.

The examples clearly show that direct application of squalene to humans should be done with great care, it is necessary to find ways for safe use of this natural triterpene.

Another picture we see for derived squalene, such as highly purified squalane has been used successfully as a component of adjuvant compositions (U.S. Pat. US 6165481, 26.12.2000). 2,3-Occidentale highly effective for lowering cholesterol levels in the blood [13].

As anticancer agents recently developed derivative of squalene, as squalamine in composition with other protiva combination with other anti-cancer agents described for the treatment of carcinoma in U.S. patent 6147060 (14.11.2000).

In Japan also, attempts were made use derivatives as squalene anti-cancer tools (publ. application JP 56113794, 1981). Used salenby ether timeintensity. Derived squalene was used as the prolongator of cytostatic agents for the treatment of carcinoma. Further developments in Japan staleproizvodit suitable as anti-cancer tools (application JP 56046808, 1988; U.S. Pat. JP 4041424, 1992), were effective only in combination with other anticancer drugs, cis-platinum (application JP 61-148118, 1986), 5-fluorouracil (application JP 60-2555720, 1985) and other

As the closest analogue is selected: for drug Chemoprophylactic effect of squalene on colon cancer" [14] as for the method: "Ozonation of terpenes and their medical use" (U.S. Pat. US 5190979, 2.03.1993). In the last described that the effect of ozone on squalene causes the latter such useful properties as antibacterial, antifungal, and other useful activity, it can be used in solar and other thermal burns.

It should be noted that the information used has convinced us that natural squalene contained in shark liver oil and vegetable oils (such as olive, soybean and others ), trebuie to minimize side effects, traditionally faced in the treatment of tumors and especially leukemias, sarcomas, and metastatic forms.

The problem is solved in that the established anticancer drug based on natural squalene, activate to acquire useful properties by heat treatment or ultraviolet (UV) radiation, infrared (IR) radiation, or exposure to reactive oxygen species. As a result of activation get activated squalene (AU), which is an oily liquid, having the following physico-chemical properties in comparison with native squalene (see tab. A).

Another aspect of the invention is a method of obtaining activated squalene, which is that natural squalene (derived from shark liver oil, or vegetable oil) are affected as follows:

A) Heat treatment: heating at t 30oWith up to 200oWith optimum 90oC for 1-6 hours.

B) the Influence of UV radiation within 1-12 hours, optimally at a wavelength of 250-350 nm for 3-6 hours.

B) exposure to IR radiation within 3-12 hours, optimally at a wavelength of 400-800 nm within the stated molecular oxygen. The processed active form of oxygen is carried out from 3 to 24 hours.

When working on the present invention we studied in detail and presents the optimal stimulation modes to achieve maximum positive results.

Naturally, for the implementation of methods of influence on squalene (activation) can be used by all known means and devices, allowing heat to irradiate UV or IR rays oily liquid, or skip through her active form of oxygen. For this no special adaptations that would not be known to the person skilled in the art, is not required.

As a result of the above manipulations received a new drug AC with high antitumor activity. The main advantage of a new drug is its selective cytotoxicity against neoplastic (tumor) cells.

To confirm the correctness of the choice of optimal modes of activation of squalene the data in the appropriate tables (see tables 1, 2, 3, 4, 5).

Table 1 shows the change in absorption upon activation of squalene treatment UV radiation at t 90oC it to the formation of a white precipitate, loss of transparency of a liquid that shows what is happening denaturation substances.

Table 2 shows the accumulation of aldehydes (%) depending on the duration of heat treatment of squalene in the 90oC. Optimal heating time can be considered 1-6 hours.

Table 3 shows the effect of heating on the density and volume of squalene when you activate it in different periods of time.

Table 4 shows the change in absorption upon activation of squalene irradiation in the UV spectrum. The table shows that the optimal level of absorption is observed when processing for 3-6 hours. With increasing time of exposure to UV radiation for more than 12 hours, the drug is activated squalene loses structure, forming a group of polymeric complexes.

Table 5 contains information about the dependence of volume and density of squalene when conducting its activation by UV-irradiation time of the UV.

Study of the cytotoxic properties of activated squalene.

The research was conducted on the lines of transformed cells in culture: L929 (fibroblast-like cells of the mouse), PC (rat pheochromocytoma), NER (hepatoma rats). In addition, we determined the growth of normal is

Investigated the influence of the drug at doses of 2,6 and 10 µl/0.3 ml medium on cell growth, DNA synthesis and protein. Used the MTT-test and standard radiometric methods of using labeled precursors - C14-thymidine and C14-leucine.

Experiments were performed in plastic 24-hole Plato (Costar) or tubes of Layton with cover glass when the initial concentration 150103cells/ml based on 6 holes (glass) 1 concentration of the drug or control.

Culture was viewed live under an inverted microscope. Cover glasses with culture recorded live after rinsing in a solution of Hanks in formalin-alcohol-acetic acid (7:2:1) and were stained with hematoxylin-eosin according to normal procedures.

The drug was dissolved in dimethyl sulfoxide (DMSO) and were added to the culture after 1 day after planting cells. The result was evaluated on the second day of growth (1 day with the drug) and 4-5 days of growth (3-4 days with the drug).

Description of the drawings.

The figure 1 presents the effect of activated squalene (I) and native squalene (II) on the line L929 tumor cells. In the figure 1 specifically shows the change in the rate of DNA synthesis (% of control) under the action of the NC to 3% from baseline.

The figure 2 shows the growth of cells under the influence of the AU (I) and NA (II) the average of 10 determinations. The figure shows that the NA significantly stimulated the growth of a tumor cell line L929.

The figure 3 shows the results of studies of the effect of SA and NA on the line of tumor cells PC 12 through 3 hours after injection. The study of rates of DNA synthesis (% of control) showed that DNA synthesis is almost completely blocked.

Figure 4 reflects the growth of cancer cell line PC 12 3 hours after injection of the AU (I) and NA (II).

The figure 5 shows the rate of protein synthesis (% of control) cell line PC 12 3 hours after injection of the AU (I) and NA (II). The figure clearly shows that the rate of protein synthesis in tumor cells significantly reduced under the action of the speaker, unlike thereof under the action of the national Assembly.

Below is a detailed description of the histological picture of the culture cell line PC 12 under the action of SA and NA.

In the control PC 12 forms a layer whose character resembles the epithelial. In the process of growing its density considerably increased. By 4-5 days of growth the monolayer consisted of quite small densely pressed to each other cells with foci monocline izual complete degeneration of the cell layer. While any live cells was absent.

The concentration of 6 μl (planting density 830103cells) on the first day of growth was caused by partial degeneration of the cell layer while maintaining individual sections or groups of spread cells. At the same time on the 4th day of growth (3-day preparation) came degeneration of cells in culture. Cell death was mainly carried out by type: pikes nuclei, size reduction, eosinophilia of the cytoplasm.

When the concentration of the drug in 2 µl of the cell layer was mainly retained in the delay of its further growth and development. The mitoses were absent.

Here is the description of the histological picture of the speaker on the cell culture L929 (fibroblast-like cells of the mouse).

At a concentration of 10 μl in 1 day was observed degeneration of culture, expressed in violation of the structure of the cell layer (monolayer) and cell degeneration. Cells were acquired rounded shape, was observed eosinophilia of the cytoplasm, the violation of the nuclear structure, pikes nuclei, vacuolization of the cytoplasm. On the 3rd day recorded the complete cell death.

At a concentration of 6 ál among monolayer was present separate dying pyknotic cell is loose arrangement of cells. At the same time in culture met maloboleznennye areas in which it is sometimes observed individual mitoses. On the 3rd day of viability was retained only certain portions of the infected cells.

In figures 6, 7, 8 shows the effect of SA and NA on the culture of tumor cells hepatoma 62 (average of 10 tests). The figure 6 shows the rate of DNA synthesis, which significantly reduced under the action of the AU in comparison with the national Assembly, especially after 24 hours. The figure 7 shows the growth of hepatoma cells 62 under the influence of the AU (I) and NA (II), which under the action of the speaker is reduced in comparison with NA. The figure 8 presents the rate of protein synthesis in the cell line hepatoma 62 24 hours after the introduction of the AU (I) and NA (II), which significantly reduced under the action of the AU in comparison with NA.

Change cytotoxicity properties of squalene depending on the time of irradiation (IR) on the subject of PC 12 cells (rat pheochromocytoma).

Cell growth after 24 hours after injection of 6 μl of the irradiated squalene presented on figure 9. The figure presents the average of 10 determinations.

As can be seen in the figure, native squalene had no significant effect on the growth of PC 12 cells. Irradiation for 1 and 3 hours is not significantly changed cytotoxic t is al tumor cell growth. However, the 18-hour period of exposure leads to a reduction of antitumor activity of the drug.

Thus, the most pronounced antitumor effect had squalene irradiated in 6-12 hours.

All these changes are confirmed by histological studies.

Change the cytotoxic properties of squalene depending on the time of heat treatment. Object cells PC-12 (rat pheochromocytoma).

The growth of PC 12 cells 24 hours after injection of 6 μl of heat-treated (90oC) squalene presented in figure 10.

As can be seen in figure 10, native squalene has no significant effect on the growth of tumor tissue. However, after an hour of heat treatment, the drug showed a pronounced cytotoxicity. Fund of cells decreased significantly (up to 69%). Remained separate sites spread cells.

Similar results showed drugs, subjected to 3 and 6-hour heat treatment. With further increase of time of heating the drug was lost cytotoxicity and the number of surviving cells was 60-70% of the control.

Thus, the most pronounced antitumor effect had squalene, podvergnutogo histological data.

Change the cytotoxic properties of squalene depending on the time of irradiation (UV). Object cells PC-12 (rat pheochromocytoma).

The growth of PC 12 cells 24 hours after injection of 6 μl of the irradiated squalene presented on figure 11. The figure 11 presents the average of 10 definitions of values (% of control).

As can be seen from the graph (and histologic analysis), native squalene has no significant effect on the growth of PC 12 cells.

After 1 hour of irradiation noted a reduction in the number of mitoses in the culture, as well as the loosening of the cell layer.

After 3-hour exposure to the drug revealed significantly more pronounced cytotoxicity. Fund of cells significantly decreased (38% compared to control); preserved only some areas of spread cells and complete absence of mitoses.

A similar picture was observed with the introduction of 6-hour exposure of the drug. After a 12-hour exposure, despite a significant degree of destruction of culture, were identified multiple mitoses, which attests that the cellular mass. It is possible that the cytotoxicity was determined by a high level of aldehydes (formaldehyde poisenelatel on the 3rd day of complete cell death patterns revealed only after 3-hour exposure. After 6-hour exposure has been preserved up to 15% of the cells.

Thus, the most pronounced antitumor effect had irradiated squalene within 3 hours.

Very interesting picture of the impact of the AU (I) and NA (II) on fibroblasts of healthy donors.

It is shown that under the influence of AU no inhibition of growth of fibroblasts (Fig.12) or DNA synthesis (Fig.13, I) while the NA reliably inhibits DNA synthesis in healthy fibroblasts (Fig.13, II). Studies on cell lines showed high antitumor efficacy of AU in the absence of cytotoxicity to healthy cells.

A further experiment was conducted on laboratory mice infected with leukemia.

The effect of the drug (AU) on the development of leukemia R.

Experiments conducted on mice - hybrids BDF1weighing 22-24 g intraperitoneally (b) grafted cell leukemia R in the amount of 106cells in the mouse.

To study the kinetics of the growth of leukemia R daily scored 2-3 mice from groups of control and treated animals were removed ascitic fluid and after repeated washing of the abdominal cavity with saline counted the total number of the tumor is the Odile per os, since 3 days after vaccination. In the control group animals received 0.1 ml of the preparation of the national Assembly, 2 times a day. Course introduction - 5 days.

The main quantitative parameters characterizing the effectiveness of chemotherapy are summarized in table 6.

The results obtained indicate that the drug AC has a pronounced antitumor effect: significantly inhibits tumor growth within 17 days of observation and increase the lifespan of mice. All control animals died on the 10th day after inoculation of tumor in the accumulation in the abdominal cavity leukemia cells is of the order of 108(PL.6), whereas in the experimental group the number of cells is reduced (under the microscope of a single cell). After 7 days in the experimental animals the number of leukemia cells was increased, but did not reach the original data.

Clinical trials of drugs.

1. Treatment of acute leukemias.

The study included 40 patients with various forms of leukemia. The age of patients from 16 to 22 years. The observation period for patients 1 year.

The treatment was performed in a hospital (3 months). Daily dose of 0.5 ml per 1 kg of body weight. The introduction of per os. The reception frequency is 4-5 times in dago effect.

The diagnosis is based on clinical cards and laboratory data (blood punctata bone marrow). As a control every 2 weeks investigated the content of blasts in paragraphs bone marrow and complete blood count.

At the same time conducted a study of biochemical parameters of blood to oversee the functions of internal organs (liver, kidney, muscle tissue).

Clinical and laboratory observations have shown that most effectively treatable lymphoblastic and plasmablastic leukemia. Intoxication symptoms were resolved within 2 weeks, although the number of blasts during this period in punctate bone marrow remained unchanged. However, in the peripheral blood was observed an increased number of erythrocytes, platelets, activity of transaminases, CPK to physiological values.

In further revealed a reduction in the number of blasts in punctate bone marrow of patients with acute leukemia (table 7).

As can be seen from the presented data, in the process of treatment the number of blasts decreases more slowly than in the treatment of chemotherapy drugs. However, the AU has no adverse reactions and complications. The composition of red and white blood is not changed.

Clinical symptoms were eliminated in the first weeks of treatment and did not disturb the patient during the entire period of observation.

Thus, as in doses of 0.5 mg/kg of body weight not less effectively eliminate the symptoms of the disease and had no toxic effects on the body of the patient. No signs of lesions of the nervous system, liver, kidneys, and also inhibit the growth of red blood and platelets characteristic of complex hormonal and chemotherapeutic drugs, amid the throng speakers were not observed. On the contrary, in the treatment process, said the restoration of previously disturbed physiological parameters.

In other forms of leukemia therapeutic effect was observed in the same volume, but in a more extended time intervals. The first clinical and paraclinical signs of remission is fixed to the end of 1 month of starting treatment. Normalization of parameters of red and white blood cells, and biochemical parameters occurred in 40-70 day interval, whereas the elimination of blasts (punctate bone marrow) recorded only after 100-120 days of treatment. As with other forms of complications and adverse reactions are not recorded.

The invention illustriou is, 3 years old, was admitted to a surgical unit 15 may 2000 in satisfactory condition with complaints of intermittent pain in the anus with irradiation in the sacral and lumbar region. Considers himself ill in the last 4 months, when for no apparent reason have any bleeding after defecation in the volume of 10-15 ml, then came pain. Effects of intestinal obstruction was not observed.

The examination of the phenomena of tumor cachexia and no intoxication. According to the General analysis of blood - moderate anemia 95 g/L.

If rektoromanoskopii identified tumor ampullares division of the rectum to 2/3 of its diameter.

Clinical diagnosis: cancer of the rectum T2N1M0. Poorly-differentiated adenocarcinoma with elements of squamous cell cancer without rogoveanu.

Within 56 days of the patient being treated with the drug as squalene, UV irradiated for 3 hours; daily doses of 20 ml or 4 ml 5 times a day).

Throughout the course of treatment the bleeding had anticipated, pain completely stopped. The phenomenon of intestinal obstruction is not increased.

To control the anticancer activity of the drug is observed to determine the degree of cytolysis of cancer cells.

A biopsy of the tumor: 14, 28, 42, 56 day.

The biopsy results:

day 14 - cytolysis more than 50% of tumor cells;

28 day - cytolysis more than 70% of tumor cells;

day 42 - cytolysis more than 90% of tumor cells;

56 day - single tumor cells.

Tumor tissue is gradually replaced by granulation tissue with foci of angiomatosis and abundance of myofibroblasts and lymphocytes among the fields which histologically observed the remains of tubular adenocarcinoma, crushed granulation tissue. The modified fabric with traces of tumor cells requires the removal.

The patient underwent operation: brunorosetta extirpation of the rectum. Extended lymphadenectomy.

The patient was discharged 10 days after surgery in satisfactory condition.

Example 2

Patient N., 64 years old, was admitted to a surgical unit 15 may 2000 complaining of mild, persistent pain in the left abdomen more below, the phenomena of coprostasis, chronic constipation.

Considers himself ill over the last 8 months, when, after a cleansing enema about chronic constipation appeared blood in the stool in the amount of approximately 30-50 ml then kerravala in the lumbar region.

Admitted to the hospital for examination and treatment.

In the study identified the tumor rectosigmoid Department of the colon with mixed type of growth with growth in the retroperitoneal tissue and the area of the iliac vessels to the left revealed metastases in the lower lobe of the left lung.

The patient during examination revealed severe pulmonary heart disease, history - myocardial infarction 1998, 1999, angina 2 functional class, circulatory insufficiency 2 degrees. In the lung - emphysema, chronic bronchitis.

Given this clinical picture and the patient's condition resolved symptomatic therapy, shows a surgical intervention only if the increase in symptoms of intestinal obstruction.

Analysis of biopsy material allowed to diagnose. Diagnosis: cancer rectosigmoidal Department of colon T2N1M1. Cancer is a complex structure with a predominance of the elements of moderate - and low differentiated adenocarcinoma and glandular squamous cancer.

From the first day of hospitalization, along with the traditional treatment, the patient received medication AU obtained by the heat treatment. At the end of the 2nd not the s region. The chair has become a regular 2-3 times a week, bleeding was not.

Biopsy was performed 1 time per month. Histological examination showed that degenerative changes are presented in the form of olizane tumor tissue, necrosis and calcification.

At the end of treatment stated the death of 90% of tumor cells.

To prevent the symptoms of intestinal obstruction after 4.5 months, the patient performed palliative surgery - partial tumor resection with removal of colostomy on the anterior abdominal wall. Metastases and analysis (histology). In the microscopic slide of the patient revealed cytolysis more than 35% of tumor cells, isolated foci of hemorrhage, necrobiosis and necrosis of the tumor tissue was not detected.

After 7 days the patient was discharged at the place of residence for outpatient treatment. Treatment activated squalene terminated. Treatment of 150 days. Inspection two months later showed that the patient's condition is stable. Pain is absent.

Example 3

Patient B., 58 years old, was admitted to a surgical unit with complaints of mild pain in the anus, recurrent bleeding after defecation, said pohodnistvo discomfort in the anus, bleeding after defecation. For outpatient screening - colonoscopy revealed a tumor on the border of the sigmoid and the rectum, with exophytic growth. Signs of intestinal obstruction is not detected.

Was admitted for further treatment in the hospital. When receiving a satisfactory condition, intoxication effects are not observed. Pain in the first days of hospital stay were stopped by parenteral introduction of narcotic analgesics up to 5-6 times a day.

Clinical diagnosis: cancer of the rectum T2N0M0. Well-differentiated adenocarcinoma with elements prestavitelyami cancer.

Within 130 days of the patient being treated with the drug as squalene, activated by heat treatment). Daily dose of 30 ml, reception 5 times a day for 6 ml.

Throughout the course of treatment of the phenomenon of intestinal obstruction was not shown, bleeding after defecation was not renewed, pain decreased significantly (up to 1 injection of non-narcotic analgesics).

To control the anticancer activity of the drug to the patient every two weeks did rektoskopiû and a biopsy of the tumor with subsequent morphological the operating morphology (130 days).

The biopsy results:

day 14 - cytolysis more than 50% of cancer cells;

28 day - more than 70%;

42 day - more than 90%;

56 day - more than 90%;

84 day - more than 90%;

130 day - more than 90%.

After treatment operation performed: brunorosetta extirpation of the rectum. Extended lymphadenectomy.

Postoperative period was unremarkable. The patient is prepared for discharge.

After a year of treatment, the patient was re-examined: 1) the pain does not bother the patient; 2) bleeding is not registered; 3) effects no obstruction; 4) signs of recurrence of the tumor no.

Example 4

Patient R. , 67 years old, was admitted to a surgical unit 15 October 2000 in the state of moderate severity, with complaints of weakness, fatigue, malaise, nausea, vomiting after eating, expressed intense pain in the abdomen without precise localization.

Ill for the last 6 months, when on a background of full health appeared nausea, General weakness, then joined the expressed painful syndrome, nekupiruemy taking non-narcotic analgesics. In the survey by place of residence revealed generalized opochina - cancerosas. During the examination in the clinic - diagnosis. Case recognized inoperable. Appointed symptomatic therapy.

When viewed in hospital - conscious, inhibited, the contact becomes reluctantly, negative to medical staff. The skin is a normal color, turgor reduced. Hypotrophy of muscles and subcutaneous tissue. Abdomen moderately increased in size, symmetrical, slightly swollen. Painful during deep palpation in the epigastrium. Liver +2.5 cm from the edge of the costal arch, tight.

The expressed painful syndrome was stopped by the introduction of narcotic analgesics (morphine, omnopon) up to 7-8 times a day. The patient noted a complete lack of appetite, food intake was accompanied by nausea and vomiting.

From the first day of hospitalization, along with the traditional treatment, the patient received the drug as. Positive results were obtained during the first weeks of treatment. Changed the character of the pain is localized in the epigastrium, became less intense, more dull and aching. With 10 days to relief of pain was enough 1-2 injections of narcotic analgesic. By the end of the first week were stopped phenomena dyspepsia (nausea and vomiting), appetite. The patient was itself absorbere arc.

Throughout treatment, the patient every two weeks was performed gastroscopy and biopsy of the tumor with subsequent morphological study on the extent of cytolysis of tumor cells detected multiple metastases biopsy.

A biopsy of the tumor: 14, 28, 42, 56, 84 day.

day 14 - cytolysis more than 30% of tumor cells;

28 day - cytolysis more than 50% of tumor cells;

day 42 - cytolysis more than 70% of tumor cells;

56 day - cytolysis more than 80% of tumor cells;

84 day - cytolysis more than 90% of tumor cells;

Conclusion: the observed phenomena beginning degradation of the tumor.

Example 5

Patient N. , 68 years old, was admitted to a surgical unit 18 October 2000 complaining of discomfort after eating, heaviness in the epigastrium, occasionally dyspeptic symptoms. Pain syndrome not.

For medical examination in September 2000 during gastroscopy revealed a tumor of the stomach. Referred for surgical treatment.

When signs of tumor cachexia and no intoxication. According to analyses of changes it is not revealed.

Belly symmetric, not swollen, participates in the act of breathing, symptoms of Rasta rorycesoja Department of the stomach T1N0M0. Well-differentiated tubular adenocarcinoma.

The patient from the first day of inpatient prescribed medication as (squalene, activated by UV-irradiation), 25 ml / day (5 times a day 5 ml).

Throughout the course of treatment every two weeks the patient was performed gastroscopy and biopsy of the tumor with subsequent morphological study on the extent of cytolysis of tumor cells. The results are shown below.

A biopsy of the tumor: 14, 28, 42 day.

The biopsy results:

day 14 - cytolysis more than 30% of tumor cells;

28 day - cytolysis more than 60% of tumor cells;

day 42 - cytolysis more than 70% of tumor cells.

After completion of the course completed operation: gastrectomy, lad.

On the 10th day after surgery the patient was discharged in good condition.

Inspection after 6 months showed that the patient's condition remains stable, signs of regressi no tumor.

Thus, activated squalene (regardless of the processing method has proven to be an effective anticancer drug, not having any adverse reactions or onlinelicensing means of Russia, 2000, pp. 1221-1223).

2. Fan Shyn-Rong et al. "Mutat. Res. Genet. Toxicol.", 1996, 3-4, 165-169.

3. Miettinen T., Vanhanen, H., "Am. J. Clin. Nutr.", 1994, 59: 356-363.

4. Kelly, G. S. "Altern. Med. Rev., l999, 4 (1), 29-36.

5. Yoder J. et al. (U.S. Pat. US 5404599, 1996).

6. Escribano, E. et al. "STP pharma sci.", 1995, 6, 461-467.

7. Ann. N.-J. Acad. Sci., 1999, 889: 193-203.

8. Lee J. S. et al. "Abstr. 10thEuropean Congress of Radiolodgy - ECR'97", Vienna, Austria, 1997, p. 961".

9. Lee J. S. et al. "J. Comput. Assist. Tomogr.", 1999, 23 (5), 730-735.

10. Michikawa M. , Yanigisawa K., "J. Neurochem", 1999, 72 (6), 2278-2285.

11. Gaj Kowska B. et al. "Exp. Toxicol. Pathol", 1999, 51 (1), 75-80.

12. Asa P., Cao Y, Garry R. F. "Exp. Mol. Pathol", 2000, 68 (1), 55-64.

13. Morand O. H. et al. "J. Lipid Res.", 1997, 2, 373-390.

14. Rao Chinthalapally V. et al. "Cancerogenesis", 1998, v. 2, 287-290.

1. The anticancer drug based on natural squalene, characterized in that it contains squalene, activated by heat treatment or ultraviolet (UV) radiation, infrared (IR) radiation, or exposure to reactive oxygen species, activated squalene (AU) is a colorless or slightly yellowish viscous liquid, having a boiling point of 238-240oWith, share 0,853-0,855.

2. The drug under item 1, characterized in that it is effective in cancer of the stomach, lung, intestine, lechaschau processing of squalene, characterized in that the treatment leads to a drug as, when this heat treatment is conducted by heating squalene at a temperature of 30-200oWith, or by exposure to UV-radiation within 1-12 hours, or exposure to IR radiation within 3-12 h, or the influence of an activated form of oxygen, selected from the group of: ozone, hydrogen peroxide, transmission of molecular oxygen.

4. The method according to p. 3, characterized in that the heating is carried out at a temperature of 90oWith, exposure to UV radiation is carried out at a wavelength of 250-350 nm for 3-6 h, the effect of IR radiation at a wavelength of 400-800 nm for 6-9 h, and the processing of the active form of oxygen - 3-24 hours

 

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1 tbl, 4 ex

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