JP2003508487A - Methods for treating cell proliferative disorders, including cancer - Google Patents

Abstract

  (57) [Summary] A method of treating a cell proliferative disorder, including a type of cancer, comprises administering a predetermined amount of a mixture of polyunsaturated fatty acids, optionally in the form of at least one polyunsaturated fatty acid solution, to a predetermined anticancer Administration with drugs, cytokines, and oleaginous lymphangiography causes an inhibition of blood supply to cancerous areas (eg, tumors). In a preferred form, the mixture is injected intra-arterially via a catheter into an artery that provides the majority of the blood supply to the tumor, which is adjacent to the tumor.

Description

Detailed Description of the Invention

[0001]   (Field of the invention)   The present invention relates to methods for treating cell proliferative disorders, including many types of cancer.

BACKGROUND OF THE INVENTION Over the last few years, efforts have been made to invent methods of treating numerous types of cancer, including cell proliferative disorders, and more specifically glioma.

[0003]   (Abbreviations used in the present invention) TNF = tumor necrosis factor. IFN = interferon. EFA = essential fatty acid. PUFA = polyunsaturated fatty acid. LA = linoleic acid. GLA = γ-linolenic acid. DGLA = dihomo-GLA. AA = arachidonic acid. ALA = α-linolenic acid. EPA = eicosapentaenoic acid. DHA = docosahexaenoic acid. CT = computed tomography. MRI = magnetic resonance imaging.

Designing methods for the selective delivery of drugs to target tissues is an established goal in the field of medicine. Generally, a drug or combination of drugs or derivative of a drug or group of drugs / groups of drugs / drugs, such that the dosage and consequently side effects may be reduced or prevented without compromising the ultimate desired effect on the target tissue or organ.
It is always desirable to selectively deliver a compound to its target. This is especially important in the case of anti-cancer drugs or anti-cancer drugs. Because, therapeutic doses or effective doses therefor for treating cancer often refer to anti-cancer agents or drugs
It is limited by the side effects of toxicity to normal healthy tissues / organs.

One method that may be employed to overcome the toxic side effects of an anti-cancer drug or anti-cancer agent is to prevent the side-effects of the anti-cancer drug or anti-cancer drug either with this method or in combination with it. By administering the resulting compound or agent. This method of approach to treating cancer provides that an anti-cancer drug or anti-cancer drug can be combined with or combined with a compound that also has a significant effect of killing cancer cells by itself. , Exhibits further significance.

It is known from the available literature that polyunsaturated fatty acids (PUFAs) are known to have cytotoxic properties against tumor cells in vitro. However, the conditions that exist during in vitro situations that affect the action of PUFAs, either alone or in combination with other additives, on tumor cells are due to various reasons.
It cannot be reproduced in living organisms.

Essential fatty acids (EFAs) are natural compounds that are present in human food but cannot be synthesized in the body. They are called essential fatty acids because they are not formed in the body and are essential for normal health. These EFAs belong to the class of fatty acids called polyunsaturated fatty acids (PUFAs). PUFAs are classified according to the short hand nomenclature, which specifies the number of carbon atoms present (chain length), the number of double bonds in the chain, and the position of the double bond closest to the terminal methyl group. To be done. The term n-x is used to describe the position of the double bond closest to the methyl group. There are four independent families of PUFAs, depending on the parent fatty acid from which they are synthesized. These are: a. N-6 series derived from linoleic acid (LA, 18: 2) b. n-3 series derived from α-linolenic acid (ALA, 18: 3) c. N-7 series derived from palmitoleic acid d. N-9 series derived from oleic acid (OA, 18: 1).

The fatty acids of the invention of the n-3 series and the n-6 series of the invention cannot be synthesized by mammals and therefore they must be taken in food and for this reason essential fatty acids Called. All EFAs are PUFAs, but all PUs
It should be emphasized that FA is not an EFA. 4 PUF
Of the A family, n-3 and n-6 are the most important. Because these and their metabolites have a wide variety of biological effects.

Both LA and ALA are thought to be metabolized by the same set of enzymes (see Figure 1). LA is, by the action of the enzyme δ-6-desaturase,
It is converted to GLA, which is then extended to form dihomo GLA (DGLA), which is a series of precursors for prostaglandins. d-6-d (δ-
The reaction catalyzed by 6-desaturase) is the rate-limiting step in the metabolism of EFA. DGLA can also be converted to AA by the action of the enzyme delta-5-desaturase. AA forms two series of precursors of prostaglandins, thromboxane, and four series of leukotrienes.

ALA is converted to EPA by d-6-d and d-5-d. EP
A forms precursors of 3 series of prostaglandins and 5 series of leukotrienes. The activities of d-6-d and d-5-d are genetically determined. d-6-
Both d and d-5-d are present in almost all tissues of the body. Conversion of EFAs to prostaglandins and thromboxane occurs via the cyclooxygenase pathway, while leukotriene formation occurs via the lipoxygenase pathway. Here, EFAs and their metabolites may have a very important effect on the host, as well as the fact that PG, TX (thromboxane) and LT (leukotriene) (which are also important biological agents). It is known to have clinical significance despite forming a precursor for the formation of (having an effect). LA, GLA, DGLA, AA, ALA, EPA and DHA are all polyunsaturated fatty acids, and only LA and ALA are essential fatty acids (EF
A). Therefore, when the term EFA is used, this is LA and AL.
A, while when the term PUFA is used, this is LA, GLA, D
Refers to GLA, AA, ALA, EPA and DHA.

US Pat. No. 5,763,484 to Horrobin (issued June 9, 1998) discloses one or more metabolites of linoleic acid and one or more metabolites of α-linolenic acid (esters, Are administered in the form of salts, amides or other derivatives that are convertible in the body) and are used to teach cancer. Horrobin
The patent also states that a human suffering from atopy, a condition caused by an abnormal immune response, may have a deficiency in the ability to convert linoleic acid to γ-linolenic acid.

US Pat. No. 5,246,726 to Horrobin et al. (September 1993)
Issued on 21st March), iron compounds and essential fatty acids (especially γ-linolenic acid, dihomo-γ-linolenic acid or eicosapentaenoic acid) are added in an amount of 1 mg to 100 gm per day.
Used to treat cancer. US Pat. No. 5,603,959 to Horrobin et al., Issued February 18, 1997, treats rheumatoid arthritis, osteoarthritis, and other disorders including Alzheimer's disease using essential fatty acids. Teach the method. In particular, in this patent to Horrobin et al., A nonsteroidal anti-inflammatory drug (NSAID) chemically linked to an essential fatty acid.
) Is used for treatment.

US Pat. No. 5,795,909 to Shashoua et al.
Issued on May 18) is cis-docosahexaenoic acid and taxane.
For the use in the treatment of cell proliferative disorders.

Hayashi Y. By Cancer et al. In Cancer Research,
"Anticer effects of free polyunsa
toured fatty acids in an oily lymph
graphical agent following intrahepati
c artial administration to a rabbi
The publication of January 15, 1992, entitled "t bearing VX-2 tumor", describes the anti-hepatic cancer effect of three free polyunsaturated fatty acids (linoleic acid, α-linolenic acid and γ-linolenic acid). Teach. This publication describes that intrahepatic administration of Lipiodol® containing free fatty acids is an effective way to deliver these three fatty acids as anti-cancer agents.

Chem Pharm Bulletin 38 (10): 2874-6 in Hayashi Y. et al. Et al., October 1990 publication, teaches the release characteristics of free polyunsaturated fatty acids from oily lymphatic contrast media. Prolonged release of α-linolenic acid from Lipiodol as a prerequisite for selective anti-cancer effects is discussed in this publication.

[0016] Kinoshita et al., In Vivo 8 (3): 371-4, May-June 1994, discloses purified linoleic acid (LA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The effect of) on breast tumors implanted in mice. Also by the author Kinoshita et al.
The number of lung metastases was investigated. Hayashi Y. J Pharmaco by et al.
Biodyn 13 (11): 705-11, November 1990 publication, examines the in vitro cytotoxicity of γ-linolenic acid on rat hepatoma cells. It was reported that the cytotoxicity of γ-linolenic acid closely correlates with the concentration of unbound (free) γ-linolenic acid. It was observed that lipid peroxidation was inhibited by serum albumin. This publication shows that the presence of albumin
It is described that the cytotoxicity of the administered free fatty acid is suppressed.

Publications September-October 1992; Nutriti by Ramesh G et al.
on 8 (5): 343-347 is the cytotoxicity of GLA, which is present in evening primrose oil, albumin, and ascites (as) in animals with ascites tumors.
It was demonstrated to be protected by other proteins present in the citric fluid). Albumin may bind to GLA and thus prevent its ability on tumor cells. This can bind to plasma albumin whenever GLA and other fatty acids are administered orally or parenterally, which in turn causes fatty acid cytotoxicity to tumor cells. It was shown to suppress.

Dr. U. N. According to Das, Nutrition Vol 6, No. 6
The 1990 / December 1990, page 429 publication of 1990 describes the cytotoxic effects of lymphokines, anticancer drugs and c-UFA. This publication lists aberrant metabolic events in most tumor cells, including: (i) a reduction in free radical production associated with a relative increase in the capacity of antioxidants; (ii) human preference. Reduced content of polyunsaturated fatty acids (PUFAs) required to induce oxidative metabolism in neutrophils and tumor cells, (iii) increased polyamines, and (iv) partially host cell proliferation, Genetic abnormalities that may be responsible for invasion and metabolism, and development of resistance to anti-cancer drugs.

This publication shows that lymphokines (eg, in the form of interferon and tumor necrosis factor) are antitumor by releasing c-UFA from cell membrane lipid pools and subsequently inducing free radical production. It was observed. Also, drugs such as vincristine increase free radical production and lipid peroxidation.

M. R. C. Naidu, U .; N. Das and A. According to Kishan,
Prostaglandins Leukotrienes and Esse
"Natural Fatty Acids": 1992 45, (181-184).
1991 publication discusses selective delivery of γ-linolenic acid (GLA) to tumor cells and intratumoral injection of GLA in the management of glioma.

Sangeetha Sagar and U.S.A. N. Das, Med. Sc
ience. Research 1993; 21: 457-459, 19.
The 1993 publication suggests that both anticancer drugs and cUFA can increase free radical production and lipid peroxidation.

Dr. U. N. Das, Cancer Letters, 56 (199).
1) 235-243 (Elsevier Scientific Publics)
Hers, Ireland Ltd) published 1991 that vitamin E, uric acid, glutathione peroxidase, superoxide dismutase and adenosine triphosphate (ATP) may block the tumoricidal activity of GLA, while iron and copper , That this activity can be enhanced.

U. N. Das et al., Prostaglandins Leukotrie.
ns and Essential Fatty Acids 1998,58
(1) The 1998 publication in 39-54 describes that a cell cycle protein called cyclin plays a crucial role in controlling cell growth.
This publication also teaches that, as the experiments show, preincubation of vincristine-resistant cells with a dose of cis-unsaturated fatty acids enhances the cytotoxic effects of vincristine.

SUMMARY OF THE INVENTION The present invention, in one aspect, teaches a method of interrupting blood supply to a tumor area to cause necrosis with highly desirable results. The present invention also provides a method of inducing an anti-angiogenic effect in a tumor area, the result of which no new blood vessels and collaterals are formed and tumor maintenance in the tumor area treated according to the present invention. do not do. The present invention, in another aspect, addresses the problem of drug delivery to target tissues and administers a mixture of selected PUFAs with other elements as described herein below. Provide a way.

Tumor cells lack phospholipase A2, an enzyme required for the release of various PUFAs from cell membrane lipids, which results in anti-tumor PGs (eg P
The production of GD2) is not refined. Moreover, tumor cells secrete excess PGE2, an immunosuppressive and mutagenic substance. Furthermore, tumor cells lack PUFAs such as GLA, AA, EPA and DHA due to the low activity of delta-6-desaturase. As a result of these metabolic changes, tumor cells can effectively block the body's defenses and survival. The present invention provides a method of causing tumor cell necrosis regardless of the known survival pattern of the tumor cells.

Anti-cancer effect of PUFAs: Tumor cells are not only poor in PUFAs, but also have a low rate of lipid peroxidation and relatively high amounts of antioxidants such as vitamin E and superoxide. Dismutase (SOD)). It is also believed that the low rate of lipid peroxidation in this cell and the resulting low amount of lipid peroxide may contribute to an increase in the mitotic process that ultimately leads to an increase in cell proliferation. Therefore,
The lack of PUFAs in this tumor cell, the presence of high amounts of antioxidants, and low amounts of lipid peroxide may contribute to the growth of tumor cells. This is supported by our studies, where it is noted that PUFAs such as GLA, DGLA, AA, EPA and DHA can reduce tumor cell proliferation. Moreover, when appropriate amounts of GLA, DGLA, AA, EPA and DHA were added to tumor cells and normal cells obtained from the American Type Culture Collection, any significant effect on normal cell survival in vitro was observed. It was also observed that only tumor cells were killed, even without. In mixed culture experiments in which both normal and tumor cells were grown together, GLA was added to AA, EPA and DH.
Although showing a more selective tumoricidal effect compared to A, these latter fatty acids are also somewhat effective. This means that GLA, DGLA, AA, EPA and D
We show that the selective delivery of HA to tumor cells could provide a new therapeutic approach in the treatment of cancer.

These in vitro results are supported by in vivo studies conducted in animal tumor models. For example, GLA, DGLA, AA, EPA and DHA, when used either in the pure fatty acid form alone or in the fatty acid-rich oil form, induce skin dermal papilloma growth in mice, hepatoma in rats and experimental animals. It is noted that it may inhibit the formation and growth of ascites tumor cells in the peritoneum. These results indicate that these fatty acids can inhibit the growth of various tumors even in vivo. In further studies, these fatty acids could selectively enhance the processes of free radical production and fat peroxidation in tumor cells, but not in normal cells, and therefore these fatty acids were found to kill the cancer. It was noted that it could have an effect.

PUFA enhances free radical production and lipid peroxidation in tumor cells
This ability of lysine is similar to the antitumor effects of lymphokines such as tumor necrosis factor (TNF) and interferon (IFN) of both α and β species. These lymphokines, also called cytokines, can induce the release of PUFAs from cell membrane lipid pools and increase the production of free radicals in this cell. Similarly, some anti-cancer agents, including but not limited to doxorubicin and vincristine, have the ability to enhance the production of free radicals and promote lipid peroxidation. Furthermore, PUFA and its products are
It can modulate the immune response and enhance the respiratory burst of neutrophils and the production of free radicals by macrophages. This evidence is further substantiated by the observation that the incidence of cancer in Eskimos is low due to the effects of their traditional diet, which is rich in EPA and DHA. Our research is based on PU
It has been shown that FA can be developed as a potential anti-cancer agent, either alone or in combination with lymphokines and traditional anti-cancer agents.

In a series of studies by the inventor, the cytotoxic effects of anti-cancer drugs such as doxorubicin, vincristine and cis-platin have been shown to be GLA, DGLA, AA, E.
It was also observed that it could be enhanced by various PUFAs such as PA and DHA. Furthermore, these fatty acids may also increase the cellular uptake of these anti-cancer drugs by tumor cells, and thus these fatty acids may enhance the anti-cancer effects of these drugs. In another similar experiment by the inventor, GLA, DGLA,
It was also observed that AA, EPA and DHA can kill TNF resistant L-929 tumor cells in vitro. Furthermore, these TNF resistant tumor cells were
Pretreatment of these L-929 cells with A., DGLA, AA, EPA and DHA sensitized them to TNF. These results indicate that not only are PUFAs able to kill tumor cells by themselves, but also various anticancer drugs, lymphokines (eg,
TNF and IFN) can enhance the cell killing effect, and also anticancer drugs and T
We show that tumor cells resistant to NF can be sensitized to the cytotoxic effects of various anticancer drugs and lymphokines.

In another set of experiments, vincristine-resistant tumor cells, KB- ^chR 8-5
It was also noted that (from now on KB-8-5 cells) can be sensitized to the cytotoxic effect of vincristine by GLA, DGLA, AA, EPA and DHA. Furthermore, when sub-optimal doses of vincristine and fatty acids are added together to these vincristine resistant cells, an optimal (ie significant) cell killing effect is produced. This suggests that vincristine and other anti-cancer compounds and PUFA enhance the cytotoxic effects of each other when added together to cancer cells. Vincristine-sensitive (KB-3-1) cells and vincristine-resistant (KB-8-5)
Fatty acid analysis of both cells of E. coli revealed that these resistant cells had low amounts of GLA, AA, EPA and DHA compared to vincristine-sensitive tumor cells, which indicates that deficiency of these fatty acids We show that it may be responsible for its resistance to the cytotoxic effects of anti-cancer drugs. This suggests that even drug resistant tumor cells can be killed by these fatty acids, since both vincristine sensitive and vincristine resistant tumor cells are easily (and to some extent) killed in vitro by various PUFAs. To do.

In yet another set of experiments, we also found that L-929 cells (referred to as TNF-resistant L-929 cells), which are resistant to the cytotoxic effects of tumor necrosis factor, also differentiated these cells. It was noted that pre-treatment with PUFAs could sensitize to the cytotoxic effects of TNF. In other words, L-929 cells that are resistant to the cytotoxic effect of TNF can be sensitized to the cytotoxic effect of TNF by PUFA. This means that PUFAs can not only kill tumor cells, but also give different tumor cells that are responsive to the cytotoxic effects of various anti-cancer drugs and lymphokines (cytokines) (eg tumor necrosis factor). Further show that it can serve as.

Human Studies: Many in vitro and animal studies suggest various compounds that can kill tumor cells, but these results are always extrapolated to the human context.
It is not possible. Many compounds that show excellent cancer killing effects in the laboratory cannot hold any promise when they are used in humans. To ascertain the possible uses of various PUFAs in humans, the present inventor has shown that clinical studies in patients with Hodgkin and non-Hodgkin's lymphoma, human cerebral glioma, primary hepatoma (hepatic cancer) and giant cell tumor of bone. The test was conducted. In all of these studies, it was observed that these PUFAs (especially GLA) could not only prevent tumor growth, but also regress tumor size to a significant extent. In the case of Hodgkin's and non-Hodgkin's cancers, PUFA, GLA are given orally, whereas in the case of human cerebral gliomas (malignant gliomas and other malignant tumors of the brain), GLA Given as an intratumoral injection. In the case of hepatoma and giant cell tumors of bone, in our study GLA was given by selective intra-arterial catheterization in combination with an oily lymphatic contrast agent as an emulsion. There were no significant side effects noted in all these studies, and all this patient responded to this treatment. In this situation, PUFAs may bind albumin and other proteins, so that when given intravenously, PFA
Important is the fact that UFAs are not available for use by tumor cells and as a result cannot exert their cytotoxic effect on tumor cells. In view of this, PUFAs (including GLA) should be delivered to patients in a manner that is readily available to tumors (tumor cells), and it is essential that they be selectively delivered to tumor cells. Is. Select if PUFAs (including GLA) are given intratumorally when made experimentally in the case of human gliomas or experimentally in the case of hepatoma and giant cell tumors of bone It is highly desirable to be delivered intra-arterially by selective intra-arterial infusion. However, in some cases of cancers such as Hodgkin's lymphoma and non-Hodgkin's lymphoma, where these tumor cells are extremely sensitive to the cytotoxic effects of PUFAs, even oral administration is observed in certain patients. It is also possible that it may be sufficient to be done. PUF
Since A can enhance the cell killing effect of anti-cancer drugs and lymphokines, a combination of PUFAs, anti-cancer drugs, lymphokines (eg, TNF and interferons or combinations thereof) can be used as a carrier agent (eg, It is desirable to administer with or without an oily lymphatic contrast agent. Further studies also revealed that PUFAs (eg GLA, DGLA and EPA) may prevent or ameliorate the side effects of anti-cancer agents (eg γ-rays and cis-platin) on bone marrow cells of mice. Therefore, PUFAs and conventional anti-cancer drugs /
When the anti-cancer agents are given together, they not only enhance their respective cytotoxic effects on tumor cells (thus producing a synergistic and / or additional effect on their ability to eliminate tumor cells), but also conventional anti-cancer agents. It has been found to lead to the elimination, reduction or amelioration of the side effects of. PUFAs can enhance the cytotoxic effects of conventional anti-cancer agents and lymphokines, leading to a significant reduction in the dose of these latter agents without compromising their ultimate advantage (ie, elimination of tumor cells or tumors). It is also possible.

In vivo PUFAs and conventional anticancer drugs / compared to in vitro results
Some of the phenomena that reduce the cytotoxic effects of anti-cancer agents include the following: a. PUFAs, when administered orally or intravenously, may bind albumin and other proteins in living organisms and may not be available for use by tumor cells.

B. The cytotoxic effect of PUFAs results from increased free radical production and lipid peroxidation in tumor cells only, but not in normal cells. The intensity of cytotoxicity is disadvantageously reduced in actual clinical trials because of the inefficient transport of this fatty acid to the target area.

C. A continuous blood supply to tissues with proliferative cell damage does not help to produce a significant amount of necrosis, especially if malignant cells increase faster than they are destroyed.

D. N. Madhavi and U. N. Published by Das "Ca
from a study reported in "Necker Letters" (June 1994)
Antioxidants such as vitamin E and superoxide anion exchanger, superoxide dismutase (SOD), PUs such as GLA, EPA and DHA
It has been found that radical generation and lipid peroxidation caused by FA can be completely inhibited. It was found that selective drug delivery to target tissues helps the efficacy of the beneficial effects of PUFAs.

In one aspect, the invention pertains to a method of inhibiting blood supply to a tumor, the method comprising the steps of: an artery that carries the major blood supply to the tumor, which artery is adjacent to the tumor. The arteries which are present), and a predetermined amount of polyunsaturated fatty acid (PUFA) LA, GLA, DGLA, AA,
Intraarterial injection in the form of a solution of at least one PUFA selected from ALA, EPA and DHA.

In another aspect, the invention pertains to a method for facilitating visualization of tumor treatment and tumor response in response to treatment, the method comprising the steps of: (a) tumor Transporting most of the blood supply to and locating the artery in proximity to the tumor; (b) obtaining an initial radiographic image of the tumor area; (c) (i) an oily lymphatic contrast agent. , (Ii) LA, GLA, DGLA, AA
, ALA, EPA, and a mixture of at least one PUFA lithium salt solution selected from DHA, into the artery; (d) a second and subsequent radiograph of the tumor area after a predetermined time. Obtaining an image; and comparing the first and second and subsequent radiographic images to assess the extent of regression of the tumor.

In another aspect, the invention pertains to a method of causing necrosis in a cancerous tumor by inhibiting blood supply to the tumor, the method comprising the steps of: (a) proximity to the tumor Arteries that carry the major blood supply to this tumor
(B) (i) oily lymphatic contrast agent; (ii) LA, GLA, DGLA, AA
Injecting a mixture of at least one essential fatty acid lithium salt solution selected from ALA, EPA and DHA into the located artery; (c) waiting for a predetermined time, and performing a radiographic study or other Assessing the extent of necrosis in this tumor by testing by means; and (d) repeating step (b) if necessary to increase necrosis.

In yet another aspect, the invention pertains to the treatment of glioma and a method of visualizing glioma regression when it responds to treatment, which method comprises: (a) a nerve Obtaining a first radiographic image of a region containing glioma; (b) (i) a sodium salt of at least one polyunsaturated fatty acid selected from LA, GLA, DGLA, AA, ALA, EPA and DHA. Injecting a solution or any other suitable salt solution, or a mixture of combinations thereof, into the glioma area; (c) a second and subsequent X-ray of the glioma area after a predetermined period of time. Obtaining a radiographic image; and comparing the first radiograph showing the glioma with the second and subsequent radiographs of the glioma area to determine the extent of remission of the glioma. Visualization and evaluation process.

In yet another aspect, the present invention uses a parenterally administered lithium salt of PUFA or an emulsion of a combination of PUFA and a given anti-cancer drug to treat a cell proliferative disorder in a mammal. Belongs to the method of treatment. Preferably, the intraarterial administration of the mixture containing PUFA (s) is via a catheter. Also, the arteries that carry the major blood supply to the tumor are referred to herein as the tumor feeding vessels.
It should be understood as synonymous with the artery supplying the eding vessel). The present invention does not lead to the formation of new blood vessels (ie, angiogenesis) due to the phenomenon that is the result of impairing blood supply.

A more detailed understanding of the present invention can be obtained from the following description of the preferred embodiments, given by way of example, and should be understood in conjunction with the accompanying drawings / drawings.

Detailed Description of Embodiments Figure 1 shows a representative known metabolic pattern of essential fatty acids, as known in the art. Essential fatty acids are precursors of eicosanoids and are important structural components of cell membranes. Essential fatty acids also provide a substrate for the production of lipid peroxidation products, which have an inhibitory effect on cell proliferation. Tumor cells have low δ-6-
It is known to have a desaturase activity, and δ-6-desaturase is
Dietary linoleic acid (LA, 18: 2, n-6) and α-linolenic acid (ALA,
18: 3, n-3) are enzymes required to desaturate their respective products. In a previous study, the inventor found that the liver cancer inducer, diethylnitrosamine (DEN) and 2-acetylaminofluorene (2-acetylam).
(2-AAF) suppresses the activities of δ-6-desaturase and δ-5-desaturase in tumor cells, resulting in low levels of γ-linolenic acid (GLA, 18: 3, n-6) and arachidonic acid. (AA, 20: 4, n-
6) and eicosapentaenoic acid (EPA, 20: 5, n-3) and docosahexaenoic acid (DHA, 22: 6, n-3). These results led the inventors and others to study the effects of various fatty acids on tumor cell survival in vitro. EFA (LA and ALA) and other PUFA (
For example, the addition of GLA, DGLA, AA, EPA and DHA) to various tumor cells in vitro showed that only the tumor cells were killed by these fatty acids without harming normal cells. This selective tumoricidal effect of fatty acids appears to be mediated by free radicals and lipid peroxides. Radiation, some anticancer drugs and cytokines (lymphokines) as well as these fatty acids
Also appear to have the ability to generate free radicals in tumor cells, thus leading to their tumoricidal effect.

Since drug resistance is a major obstacle in the clinical treatment of cancer, and PUFA has a selective tumoricidal effect, the inventors have determined that PU against drug resistant tumor cells.
The effect of FA and the effect of their regulation on the action of anti-cancer drugs was studied.

In the above situation, in addition to causing tumor cell drug resistance reversal by administration of polyunsaturated fatty acids, in the present invention, the mode of targeting cancerous tissue depends on the effect and rate at which necrosis can occur. It turns out that it is very critical. More specifically, throughout the present invention, tumors are delivered to a tumor site by intra-arterial delivery of a selected mixture of a salt of a given polyunsaturated fatty acid and a given anti-cancer agent to the tumor site by using the artery closest to the tumor site. It will be appreciated that a very beneficial and hitherto unknown effect is achieved in that it impairs the blood supply to the site.

In a clinical study conducted by the present inventor, it was asserted that this inhibition of blood supply was sufficient to block the blood supply to the tumor site with little time lag. In other cases, a consistent suppression of blood supply to the tumor area was observed, but relatively gradual.

In the following description of the observations and clinical studies on the treatment of patients with hepatocellular carcinoma, giant cell tumors of the scapula and giant cell tumors of the femur, PUFA salt was injected into arteries near the tumor area. The unexpected beneficial effects of inhibition of the blood supply to the tumor area and inhibition of the normal supply of its major blood supply to the tumor caused by intra-arterial injection of the mixture may be revealed.

FIG. 2 shows an actual radiographic image of a giant cell tumor of the scapula of a human patient. The tumor is shown at 21 in FIG. The major part of the blood supply to the tumor occurs via the subclavian artery, indicated at 22. The subclavian artery continues as the axillary artery, then the axillary artery as the brachial artery, which supplies the brachial blood. In this case, the subclavian artery 22 will be the major supplier of blood supply to the tumor 21 and will therefore be the selected PUFA.
An intraarterial injection of a mixture of salt and an oily lymphatic contrast agent was made into the subclavian artery 2 via a catheter 23. FIG. 3 shows the actual radiographic image of the tumor shown in FIG. 2 after only a short time, where an inhibition of the blood supply can be observed.
In fact, the present inventor has shown that further administration of this mixture is not readily possible or necessary in clinical studies, because of the rapid response after intra-arterial injection the blood pathways are rapidly blocked. I noticed.

FIG. 4 shows an X-ray image of hepatocellular carcinoma, which is a tumor of the liver. Intra-arterial administration of a mixture of a lymphatic contrast agent and a selected PUFA salt results in hepatic artery 42 via the celiac artery (coeliac axis), which was determined to be the major supplier of blood supply to the affected area. It was done by using a catheter inserted into the. Celiac artery
As is known, it is the major junction of the abdominal aorta from which the major blood vessels, including the arteries of the liver for supplying the liver, originate. The radiographic image of FIG. 5 was taken after 4 weeks. From the image of FIG. 5 it can be seen that the dispensed lymphatic contrast agent is visible in the area of the tumor and the blood supply to the tumor area is reduced. As previously described, exposure of healthy cells and tissues to PUFA or any form of its salt derivative does not adversely affect the condition or integrity of healthy tissues. The blood supply is due to unknown reasons, selected PU in oily lymphatic contrast media.
After intraarterial administration of a mixture of FA salt and any given anti-cancer drug, there is a gradual decrease to the point of complete block. In fact, the experiment, without the addition of any anti-cancer drug,
It was revealed that the blood supply was cut off using intra-arterial administration of PUFA alone.
The presence of an oily lymphatic contrast agent as a carrier visualizes the distribution and retention of the lymphatic contrast agent, indicating that PUFA salts and anticancer drugs actually arrived at the target tissue via this carrier. . An undesirable alternative for achieving massive necrosis of cells in the tumor area is surgical removal of the malignant liver, which
Very difficult and dangerous, and because these tumors are usually very vascular, they can often result in rapid bleeding during or after surgery, which ultimately leads to patient death. cause.

6 and 7 show radiographic images of a giant cell tumor 61 of bone in a human femur near the area of the knee joint (known as the popliteal fossa). Popliteal artery 62, vein (not shown),
And nerves (not shown) cross the popliteal area. Bone tumors in the popliteal area
If physically removed by ablation, this mutates the patient (mut
Ilating) surgery. By using the method of the preferred embodiment of the present invention, blood supply to the popliteal area is inhibited to the point of killing tumor cells, so that bone has the opportunity to regrow. In the cases shown in Figures 6 and 7, the preferred embodiment of the method of the present invention was used to inhibit blood supply to the tumor area and result in massive necrosis in the tumor area resulting in healthy bone growth. Gave an opportunity to recover.

8-13 show radiographic images of a computed tomography scan over time of the abdomen of a patient without contrast, showing hepatocellular carcinoma during the course of treatment with the invention.

FIG. 8 shows the diseased area (ie liver) marked prior to administration of the mixture prepared according to a preferred embodiment of the present invention. The table below shows the time course after which the images of several figures were taken: Figure 8-One day before administration of the mixture Figure 9-One day after injection of the mixture Figure 10-Two days after injection of the mixture Figure 11-Four days after injection of the mixture Figure 12-3.5 weeks after injection of the mixture Figure 13-4.5 weeks after injection of the mixture.

FIG. 9 shows a CAT scan image almost immediately after administration of the mixture. FIG. 10 shows a CAT scan image taken less than one day after the images of FIGS. 8 and 9. Figure 10
It can be seen that the oily lymphatic contrast agent is visibly distributed and is seen as a whitish substance. In Figure 11, a response to treatment can be observed. In Figures 12 and 13, this image shows remission of liver tissue and recovery to normal approximately 27 days after the start of treatment. Generally described in combination with the illustrations in Figures 2-13 for intraarterial injection of a mixture of salts of a given PUFA (with or without a selected anti-cancer agent / drug) in combination with a lymphatic contrast agent / carrier. Although examples of patient treatments are given above, the inventor has found promising results even in situations where the mixture was taken in an oral manner (eg, as a capsule). As described in more detail herein below, the combination of two or more methods of ingesting a mixture (ie oral and parenteral) has shown very encouraging results.

One aspect of the present invention resides in the preparation of a combination / composition for the treatment of cancer, wherein one or more of LA, GLA, DGLA, AA, ALA, EPA and DHA comprises TNF and Conventional anti-cancer drug containing lymphokines such as interferon /
It is administered with the drug in a suitable agent for the delivery of oily lymphatic contrast agents or these compounds, or in the absence of these lymphatic contrast agents or other agents; irradiation may be included if desired. PUFA is a daily dose of 0.5 mg to 50 g, along with appropriate doses of conventional anti-cancer drugs (eg, vincristine, doxorubicin, L-asparaginase, cis-platin, busulfan, etc.) and the required dose and stage of the disease. Depending on the daily dose of 1 mg to 50 g / week / month, and dose of 1 μg to 100 mg per day (which can be 1000 units to 10 million units for TNF) of lymphokines (e.g., TNF (α Or γ type) and / or with or without the addition of interferons (alpha or γ type), as may be sometimes determined. The combination of PUFA, conventional anti-cancer drug, lymphokine and oily lymphatic contrast agent may be administered by any one or different route, at the same time or at different times and intervals for each administration or combination (eg oral, Parenteral (intra-arterial injection, topical, intra-anal as suppository, intra-vaginal route,
Or the guidance of a suitable device (eg radiographic guidance (X-ray), CT guidance or MR)
I induction, but is not limited to these), or by selecting local injection directly into the tumor bed by stereotactic induction))). The daily dose of these compounds may be one or more long-acting, daily, weekly, monthly, or some other suitable time interval as is sometimes determined depending on need. The administration of a preparation or a depot preparation cannot be excluded. Fatty acids (PUFAs) can be present in any physiologically acceptable form, including but not limited to glycerides, esters, free acids, amines, phospholipids or salts. Traditional anti-cancer drugs may be used by themselves or in PUFA (alone or in combination (eg GLA alone or GLA and AA, LA, DGLA, ALA, E.
PA or DHA))). Similarly, TNF and IFN
Lymphokines such as can be given on their own or with PUFAs.
For intra-arterial infusion or administration of LA, GLA, DGLA, AA, ALA, EPA and / or DHA, these may be as such or as oily lymphatic contrast agents or other suitable solutions which may be given parenterally. May be given in combination with (but not limited to), or dissolved in these lymphatic contrast agents or solutions, or with these lymphatic contrast agents or solutions. All of these PUFAs, conventional anti-cancer agents, lymphokines and lymphatic contrast agents, either individually or in combination, or all at the same time or at different time intervals on either the same day / week / month, are all taken together. Alternatively, they may be given separately, with the same route or different routes if the situation requires.

(Example) 1. Rigid made by accepted standards or morphologies or methods (
Here, PUFAs are microencapsulated) or soft gelatin capsules (where the fatty acids are present in the form of oils), which at the above-mentioned doses are conventional anti-cancer drugs or It is given to people with cancer in combination with lymphokines.

2. Hard or soft gelatin capsules made by conventional methods, wherein the fatty acid and the anti-cancer drug are incorporated together in the same capsule and administered to a person with cancer.

3. In particular, human brain glioma or any other tumor (eg, bladder cancer, esophageal cancer, etc.) accessible by any route by use of a flexible fiber optic scope (eg, bronchoscope, urethroscope, hysteroscope, etc.). Appropriate dose (0.5 m), either alone or in combination, for the treatment of lung cancer, breast cancer, etc.
g / day to 50 mg / day) pure LA, GLA, DGLA, AA, ALA, E
As an intratumoral preparation of PA and DHA. In the case of head and neck cancer, the fatty acids are either by direct intra-arterial route or by selective catheterization of tumor-delivering vessels (either by the femoral, respiratory or carotid route). Is administered in. PUFAs are given to these patients daily, weekly, or monthly, or for and when needed, depending on the need for the treatment and the patient's response.

4. The PUFAs, either alone or in combination with an anti-cancer drug / lymphokine, selectively select the oily lymphangiographic agent or any other suitable agent, all in admixture, in combined form, or in a combined form. Intraarterial infusion or injection into the tumor-delivering vessel by the femoral, respiratory or carotid route or any other suitable route, or a combination thereof (eg, GLA + any conventional anticancer drug or drug + Lymphokines (eg, TNF and / or interferons), LA / GLA / DGLA / AA / ALA / EPA / DHA all alone or in combination + conventional anticancer drugs + lymphokines (eg, TNF and / or
Or IFN) + lymphatic contrast agent, oily lymphatic contrast agent in combination, or in combination, or emulsified or mixed, LA / GLA / DGLA / AA /
ALA / EPA / DHA, LA / GLA / DGLA / AA / ALA / EPA / D
HA alone or their combination of oily lymphatic contrast agents (as a mixture or emulsion, or as a conjugate and other combinations thereof).
The preparation can be administered daily, weekly, or monthly, or at other suitable time intervals.

5. A topical preparation of PUFA, either alone or in combination, with a conventional anticancer drug or lymphokine (eg, TNF / IFN or other suitable lymphokine) in a suitable vehicle, Where the daily dose (0.5u
g-100 mg) is applied topically to primary skin cancers including Kaposi's sarcoma, and / or conventional anti-cancer drugs and / or lymphokines are given either orally or parenterally.

According to different embodiments of the methods of the invention described above, (i) PUFA or cis-EFA (the essential fatty acids described herein are also referred to as cis-EFA when in the cis configuration; Because of these structures, referred to as cis-fatty acids), albumin and other proteins are administered when administered intra-arterially to a patient, or else in a combination of intra-arterial and oral administration. There is less chance of binding to this fatty acid. Consequently, PUFAs so administered using the present invention are more available for uptake by tumor cells.

(Ii) Due to the effective delivery of PUFAs to the tumor site, as previously described herein, PUFAs and administered anti-cancer agents (drugs or cytokines,
Or a combination thereof) increases the intensity of cytotoxic action. Thus, the present invention is used to relatively better enhance free radical production and lipid peroxidation in tumor cells, thereby promoting a greater degree of necrosis.

(Iii) Inhibition of blood supply to the tumor area by the method of the present invention prevents cell growth in the tumor area, which in turn allows healthy tissue to grow in situ.

(Iv) Otherwise, inhibition of radical production and peroxidation of PUFA-generated lipids caused by vitamin E and superdismutase results in intra-arterial PUFA delivery to the tumor site via the proximal artery. Due to the method of transport, it is reduced by the method of the invention.

As mentioned above, intraarterial administration concurrently with oral administration of a predetermined dose of PUFA of a mixture of PUFA, anticancer drug, and selected cytokines is also within the scope of the invention. All such changes are considered to be within the scope of the invention.

Mammalian Application: Although the above examples relate to humans, it is believed that the methods and mixtures of inhibiting blood supply of the present invention are equally applicable to other mammals.

Equivalents The present invention has been particularly shown and described with reference to preferred embodiments of the invention, but various changes in form and detail are defined by the appended claims. It will be understood by those skilled in the art that the present invention can be made without departing from the spirit and scope of the present invention. For example, sodium and calcium salts are considered equivalent to each other. The image processing technology referred to herein is based on CA
It is intended to include T, MRI, X-ray and other possible image processing methods. One of ordinary skill in the art would be able to recognize or ascertain many equivalents to the particular embodiments of the invention specifically described herein using only routine experimentation. Such equivalents are intended to be included within the scope of the appended claims.

[Brief description of drawings]

[Figure 1]   FIG. 1 shows the structural metabolism of essential fatty acids.

FIG. 2 shows a radiographic image of a giant cell tumor of the human scapula before being treated by the preferred method of the present invention.

FIG. 3 shows a radiographic image of a giant cell tumor of human scapula after being treated by the preferred method of the present invention.

FIG. 4 shows a radiographic image of hepatocellular carcinoma in a human patient prior to being treated by the preferred method of the present invention.

FIG. 5 shows a radiographic image of hepatocellular carcinoma in a human patient after having been treated by the preferred method of the present invention.

FIG. 6 shows a radiographic image of a giant cell tumor in the lower end of the femur (near the knee joint region) prior to treatment with the present invention.

FIG. 7 shows a radiographic image of a giant cell tumor in the lower end of the femur (near the knee joint region) after treatment with the present invention.

8 to 13 show CAT scan images of the abdomen of a human patient with hepatocellular carcinoma over time during the course of treatment with the present invention.

FIGS. 8-13 show CAT scan images of the abdomen of a human patient with hepatocellular carcinoma over time during the course of treatment with the present invention.

FIGS. 8-13 show CAT scan images of the abdomen of a human patient with hepatocellular carcinoma over time during the course of treatment with the present invention.

8 to 13 show CAT scan images of the abdomen of a human patient with hepatocellular carcinoma over time during the course of treatment with the present invention.

FIGS. 8-13 show CAT scan images of the abdomen of a human patient with hepatocellular carcinoma over time during the course of treatment with the present invention.

FIGS. 8 to 13 show CAT scan images of the abdomen of a human patient with hepatocellular carcinoma over time during the course of treatment with the present invention.

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Claims (22)

[Claims] 1. Use of a solution of at least one polyunsaturated fatty acid in the preparation of a medicament for selectively reducing the blood supply to at least part of a tumor, said selective reduction comprising: Locate a proximal artery that carries blood to at least a portion of the tumor, and administer a therapeutically effective amount of the solution to the artery by intraarterial injection; and a lymphangiographic agent by intraarterial injection A use, wherein the arterial and blood supply can be visualized by an angiogram, and thereby the blood supply is selectively reduced. 2. Use of a lymphatic contrast agent in the preparation of a medicament for selectively reducing the blood supply to at least a portion of a tumor, said selective reduction comprising: at least a portion of said tumor. The proximal artery carrying blood to the and administering a therapeutically effective amount of a polyunsaturated fatty acid solution to the artery by intra-arterial injection; and administering the lymphatic contrast agent to the artery by intra-arterial injection Use, wherein the arteries and the blood supply can be visualized by an angiogram, and thereby the blood supply is selectively reduced. 3. Use of a solution of at least one polyunsaturated fatty acid and a lymphatic contrast agent in the preparation of a medicament for selectively reducing blood supply to at least a portion of a tumor, said selective lymphatic contrast agent. The reduction is as follows: locating the proximal artery that carries blood to at least a portion of the tumor; and selecting by administering a therapeutically effective amount of the solution and lymphatic contrast agent to the artery by intraarterial injection. Use, wherein the arteries and the blood supply can be visualized by an angiogram, and thereby the blood supply is selectively reduced. 4. Use according to claim 2 or 3, wherein the lymphatic contrast agent is conjugated to the polyunsaturated fatty acid. 5. The treatment comprises the steps of: observing the passage of the lymphatic contrast agent into the proximal artery and the tumor by an angiogram; by at least partial vessel occlusion in the tumor. 4. The use of any of claims 2 or 3, further comprising: determining when the blood supply is reduced; and stopping the injection. 6. The method of claim 5, wherein the therapeutically effective amount is an amount effective to cause at least partial occlusion of blood vessels within the tumor as determined by the angiogram. Use as stated. 7. The therapeutically effective amount is during the perfusion of the artery with the solution.
Use according to claim 5, which is sufficient to cause an occlusion of the artery. 8. The therapeutically effective amount is between 0.5 mg and 50 gm,
Use according to claim 5. 9. The use according to any one of claims 1 to 8, wherein the polyunsaturated fatty acid is an essential fatty acid. 10. The essential fatty acid is selected from the group consisting of γ-linolenic acid, arachidonic acid, docosahexaenoic acid, eicosapentaenoic acid, di-homo-γ-linolenic acid, α-linolenic acid, and linoleic acid. Use according to claim 9. 11. The polyunsaturated fatty acid is administered in the form of a salt selected from the group consisting of lithium salt, sodium salt, and calcium salt.
8. Use according to any one of 8. 12. The method according to claim 1, wherein the polyunsaturated fatty acid is in the form of a fatty acid derivative selected from the group consisting of glycerides, esters, free acids, amides, phospholipids and salts. Use as stated. 13. The tumor according to claim 1, wherein the tumor is a cancerous tumor.
Use as described in section. 14. The use according to claim 13, wherein the tumor is a glioma. 15. The tumor is a hepatoma, lung cancer, colon cancer, breast cancer, ovarian cancer,
The use according to claim 13, which is selected from the group consisting of kidney cancer, skin cancer, Kaposi's sarcoma, esophageal cancer, gastric cancer, leukemia, and lymphoma. 16. The method of claim 1 wherein the tumor is caused by a cell proliferation disorder.
The use according to any one of 1. 17. The method further comprising injecting a therapeutically effective amount of a compound selected from the group consisting of tumor necrosis factor, anticancer drug, and lymphokine, together with or separately from the polyunsaturated fatty acid. Item 9. The use according to any one of items 1 to 8. 18. The use according to claim 17, wherein the lymphokine is selected from the group consisting of alpha interferon and gamma interferon. 19. The polyunsaturated fatty acid shared with a pharmaceutical agent selected from the group consisting of vincristine, adriamycin, doxorubicin, cyclophosphamide, cis-platinum, L-asparaginase, procarbazine, camptothecin, taxol and busulfan. It couple | bonds, The any one of Claim 1-8
Use as described in section. 20. PUFA or PUF in combination with a lymphatic contrast agent
A pharmaceutical composition comprising salt A. 21. The pharmaceutical composition of claim 20, wherein the lymphatic contrast agent is covalently attached to the PUFA. 22. A group consisting of tumor necrosis factor, anticancer drug, lymphokine, α interferon, γ interferon, vincristine, adriamycin, doxorubicin, cyclophosphamide, cis-platinum, L-asparaginase, procarbazine, camptothecin, taxol and busulfan. 21. The pharmaceutical composition according to claim 20, further comprising a compound selected from