Compositions and methods for enhancing cancer radiotherapy

Abstract

The name of the invention is compositions and methods for enhancing cancer radiotherapy. A nutritional supplement comprising fish oil and selenium has been determined to enhance radiotherapy of tumors in a synergistic manner. The co-therapy of such supplements and radiotherapy results in a significantly improved reduction in tumor volume in animal models and reduces the common side effects, such as weight loss, inflammation and gastrointestinal tract damage, relative to radiotherapy alone. It has also been found that this co-therapy can reduce the incidence of metastasis from a primary implant site and modulate the expression of genes associated with apoptosis in tumor cells. Formulations of such supplements with good tolerance and palatability are also provided.

Description

[0001] This application is a divisional application. The application date of the original application is June 13, 2018, the application number is 201880047692.1, and the name is “Compositions and methods for enhancing cancer radiotherapy”.

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 519,087, filed June 13, 2017. These and all other referenced external materials are incorporated herein by reference in their entireties. When a definition or use of a term in a reference incorporated by reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein controls. Technical Field

[0003] The field of the invention is cancer radiotherapy. Background Art

[0004] The background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication explicitly or implicitly referenced is prior art.

[0005] Radiation therapy and chemotherapy regimens used to treat cancer can significantly benefit patients, but may be ineffective or less effective for some cancers. In addition, both radiation therapy and chemotherapy are associated with significant side effects, including nausea, weight loss, hair loss, gastrointestinal damage, and skin irritation.

[0006] Attempts have been made to enhance the effectiveness of radiotherapy. For example, gold nanoparticles that have been modified to target tumor cells have been used to enhance radiotherapy (Yang et al., ACS Nano, 2014, 8(9):8992-9002). All publications herein are incorporated by reference to the extent that each individual publication or patent application is expressly and individually incorporated by reference. When the definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of the term provided herein, the definition of the term provided herein applies, and the definition of the term in the reference does not apply. Similarly, COX-2 inhibitors have been used to selectively sensitize tumor cells to radiation (Choy and Milas, J. Natl Cancer Inst (2003) 95(19):1140-1452). However, this approach may have selectivity issues and may not be effective for all types of tumors. To date, attempts to reduce the side effects of radiotherapy have mainly been to divide the total radiation dose into multiple smaller radiation doses (leaving time between the two to allow recovery), use shielding to target the tumor, and determine the tumor boundary and the positioning of the radiotherapy to the site. Unfortunately, this approach may not adequately treat all tumor cells.

[0007] Attempts have also been made to enhance the effects of chemotherapy. Some studies have shown that consuming fish oil can improve the effects of chemotherapy, but other studies have shown that fish oil may interfere (Daenen et al., JAMA Oncol (2015) 1(3):350-358). Attempts have also been made to formulate chemotherapeutic agents into nanoparticles (Xu et al., Coll. Surf. B: Biointerfaces (2006) 48(1):50-57). However, it is not clear whether all chemotherapeutic drugs are suitable for such reformulation. Co-delivery of chemotherapeutic drugs with siRNA designed to interfere with multidrug resistance has also been studied. However, such siRNAs are sequence-specific and may not be suitable for certain tumors.

[0008] Reducing the side effects of chemotherapy is often directed towards providing symptom relief. For example, antiemetics and dietary changes, such as avoiding certain foods and eating smaller, more frequent meals, can be used to alleviate nausea. Unfortunately, such approaches are not always effective. In some cases, chemotherapeutic agents with reduced toxicity are selected to alleviate side effects, but such agents may also have reduced effectiveness against tumor cells.

[0009] Therefore, there remains a need for safe and effective compositions and methods for enhancing the effectiveness and / or reducing the side effects of cancer radiotherapy and / or chemotherapy. Summary of the Invention

[0010] The present subject matter provides compositions and methods for synergistically enhancing and improving radiation therapy of tumors through the use of nutritional supplements comprising fish oil and selenium.

[0011] One embodiment of the present inventive concept is a method of treating a tumor by applying a radiotherapy regimen to a patient in need of treatment while simultaneously providing the patient with a nutritional supplement comprising fish oil and selenium (e.g., as shown in Table 1) in amounts that provide a synergistic effect in reducing tumor volume or weight. In some embodiments, the nutritional supplement is provided to the patient prior to commencing radiotherapy. Such a nutritional supplement can be formulated such that two or more components of the supplement are provided in amounts as described in Table 1.

[0012] Another embodiment of the present inventive concept is a method of reducing side effects by providing a patient with a formulated nutritional supplement comprising fish oil and selenium (e.g., as in Table 1) in an amount that reduces the side effects of a radiation therapy regimen. In some embodiments, the nutritional supplement is provided to the patient prior to commencing radiation therapy. Such a nutritional supplement can be formulated such that two or more components of the supplement are provided in amounts as described in Table 1.

[0013] Another embodiment of the present invention is a method of regulating the expression of genes (e.g., angiogenesis-related genes, apoptosis-related genes, etc.) in a tumor by providing a nutritional supplement to a tumor or an animal having a tumor, wherein the nutritional supplement comprises fish oil and selenium (e.g., as in Table 1), and wherein the nutritional supplement is provided in an amount that regulates gene expression in the tumor. In some embodiments, the nutritional supplement is provided to the patient before and during radiotherapy; in other embodiments, such a supplement is provided during or at the start of radiotherapy application. In a preferred embodiment, the nutritional supplement is formulated so that two or more components in the supplement are provided in an amount as described in Table 1.

[0014] Another embodiment of the present inventive concept is a method of reducing tumor metastasis by providing a nutritional supplement comprising fish oil and selenium (such as in Table 1) to a patient with a metastatic tumor, wherein the nutritional supplement is provided in an amount that reduces the metastatic activity of the tumor. In some embodiments, the nutritional supplement is provided before or at the start of radiotherapy, and can be provided throughout the course of radiotherapy. In a preferred embodiment, the nutritional supplement is formulated such that two or more components of the supplement are provided in amounts as described in Table 1.

[0015] Another embodiment of the present inventive concept is a method of reducing angiogenesis in a tumor, comprising providing a nutritional supplement comprising fish oil and selenium (e.g., as in Table 1) to a patient having a tumor in combination with radiotherapy, wherein the nutritional supplement is provided in an amount that reduces angiogenic activity in the tumor. In some embodiments, such a nutritional supplement is provided concurrently with the initiation of radiotherapy and can be provided to the patient during the application of radiotherapy. In a preferred embodiment, the nutritional supplement is formulated such that two or more components of the supplement are provided in amounts as described in Table 1.

[0016] Another embodiment of the present inventive concept is a method for reducing the incidence of cancer stem cells in a tumor by providing a nutritional supplement comprising fish oil and selenium (such as in Table 1) to a patient suffering from a tumor, wherein the nutritional supplement is provided in an amount that reduces the incidence of stem cells in the tumor. In a preferred embodiment, the nutritional supplement is provided concurrently with the administration of radiation therapy. The nutritional supplement can be formulated such that two or more components of the supplement are provided in the amounts described in Table 1.

[0017] Various objects, features, aspects and advantages of the present subject matter will become more apparent from the following detailed description of preferred embodiments and the accompanying drawings, in which like reference numerals represent like components. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 : Figure 1A treatment regimen is described in which nutritional supplements are provided 7 days before radiation therapy or concurrently with tumor cell implantation. Mice are sacrificed 21 days after tumor cell implantation.

[0019] Figure 2A and 2B : Figure 2A Changes in body weight over time in each treatment group are depicted. Figure 2B Figure 2. Body weight changes in each treatment group over the course of the study. Note that body weights are reported after removal of the primary tumor mass.

[0020] Figures 3A to 3D : Figures 3A to 3D A typical synergistic effect of co-treatment with radiotherapy and supplements containing fish oil and selenium on tumor volume and weight was shown. Figure 3A Shown are the changes in tumor volume in each treatment group over time. Figure 3B Provided as shown Figure 3A A detailed view of the first 11 days of treatment is shown. Figure 3C A bar graph of tumor weights for each treatment group is provided. Figure 3D Photographs of exemplary tumors from each treatment group are provided.

[0021] Figures 4A to 4G : Figure 4A : Effects of radiotherapy, supplements containing fish oil and selenium, and combination therapy on gastrocnemius muscle mass 21 days after tumor cell injection. Figure 4B : Effects of radiotherapy, NutraWell supplementation, and combination treatment on muscle mass in soleus 21 days after tumor cell injection. Figure 4C : Effects of treatment with radiation, NutraWell supplements, and combination therapy on lung (including metastatic tumor) weight 21 days after tumor cell injection. Figure 4D : Effects of treatment with radiation therapy, NutraWell supplements, and combination therapy on liver (including metastatic tumor) weight 21 days after tumor cell injection. Figure 4E : Effects of treatment with radiation therapy, NutraWell supplementation, and combination therapy on spleen (including metastatic tumors) weight 21 days after tumor cell injection. Figure 4F Quantitative results are provided from a study of lung metastases in animal subjects treated with supplements containing fish oil and selenium and / or radiation therapy. Figure 4G Results are shown of a study of Ki-67 expression in primary tumor sites and metastatic sites in animal subjects treated with supplements containing fish oil and selenium and / or radiotherapy.

[0022] Figures 5A to 5E : Figure 5A: The therapeutic effects of radiation therapy, NutraWell supplementation, and combination therapy on platelet counts 21 days after tumor cell injection. Figure 5B : The therapeutic effects of radiation therapy, NutraWell supplementation, and combination therapy on white blood cell counts 21 days after tumor cell injection. Figure 5C : Effects of radiation therapy, NutraWell supplementation, and combination therapy on lymphocyte counts 21 days after tumor cell injection. Figure 5D : Effects of radiation therapy, NutraWell supplementation, and combination therapy on granulocyte counts 21 days after tumor cell injection. Figure 5E : The therapeutic effects of radiotherapy, NutraWell supplementation, and combination therapy on neutrophil / lymphocyte percentages 21 days after tumor cell injection.

[0023] Figure 6A and 6B : Figure 6A : The therapeutic effects of radiotherapy, NutraWell supplementation, and combination therapy on serum albumin 21 days after tumor cell injection. Figure 6B : Effects of radiation therapy, NutraWell supplementation, and combination therapy on serum creatinine 21 days after tumor cell injection.

[0024] Figure 7A and 7B : Figure 7A : The therapeutic effects of radiotherapy, NutraWell supplementation, and combination therapy on serum IL-6 21 days after tumor cell injection. Figure 7B : The therapeutic effects of radiotherapy, NutraWell supplementation, and combination therapy on serum IL-1β 21 days after tumor cell injection.

[0025] Figures 8A to 8G : Figure 8A : The therapeutic effects of radiation therapy, NutraWell supplementation, and combination therapy on VEGF gene expression in implanted tumors 21 days after tumor cell injection. Figure 8B : The therapeutic effects of radiotherapy, NutraWell supplementation, and combination treatment on BAX gene expression in implanted tumors 21 days after tumor cell injection. Figure 8C : The therapeutic effects of radiation therapy, NutraWell supplementation, and combination therapy on Bcl-2 gene expression in implanted tumors 21 days after tumor cell injection. Figure 8D : The therapeutic effects of radiation therapy, NutraWell supplementation, and combination treatment on caspase 3 gene expression in implanted tumors 21 days after tumor cell injection. Figure 8E: The therapeutic effects of radiotherapy, NutraWell supplementation, and combination treatment on BAX gene expression in the lung 21 days after tumor cell injection. Figure 8F : The therapeutic effects of radiation therapy, NutraWell supplementation, and combination treatment on Bcl-2 gene expression in the lung 21 days after tumor cell injection. Figure 8G : Therapeutic effects of radiotherapy, NutraWell supplementation, and combination treatment on caspase 3 gene expression in the lung 21 days after tumor cell injection.

[0026] Figure 9A and 9B : Figure 9A A test protocol is described in which mice received radiation therapy on days 8, 10, and 12, and nutritional supplementation was initiated at the start of radiation therapy. Mice were sacrificed on day 14 or 24 after tumor cell implantation. Figure 9B :Derived from Figure 9A Treatment groups for the regimens shown in .

[0027] Figures 10A to 10C : Figure 10A :In Figure 9A Serum albumin concentrations on days 14 and 24 after tumor cell implantation in mice treated as shown. Figure 10B :In Figure 9A Lymphocyte counts on days 14 and 24 after tumor cell implantation in mice treated as shown. Figure 10C :In Figure 9A N / L ratios on days 14 and 24 after tumor cell implantation in mice treated as shown in FIG.

[0028] Figures 11A to 11D : Figure 11A :In Figure 9A VEGF expression in tumor masses 24 days after tumor cell implantation in mice treated as shown in FIG. FITC indicates VEGF-specific staining. Figure 11B :In Figure 9A VEGF expression in the lungs (metastases) 24 days after tumor cell implantation in treated mice shown in FITC indicates VEGF-specific staining. Figure 11C :In Figure 9A EGFR expression in the lungs (metastasis) 24 days after tumor cell implantation in treated mice shown in FITC indicates EGFR-specific staining. Figure 11D :In Figure 9A EGFR expression in tumor masses 24 days after tumor cell implantation in mice treated as shown in Figure 1. FITC indicates EGFR-specific staining.

[0029] Figure 12:In Figure 9A CD31 (cancer stem cell marker) expression in tumor masses and lung tissue (metastasis) 24 days after tumor cell implantation in treated mice shown in FITC indicates CD31-specific staining.

[0030] Figure 13A and 13B : Figure 13A Shown in Figure 9A Presence of H1F1-α (hypoxia marker) protein in tumor masses and lung tissue (metastasis) 24 days after tumor cell implantation in treated mice shown in FITC indicates H1F1-α specific staining. Figure 13B Shown are the results of gene expression studies of tumor samples from subjects treated with nutritional supplements comprising fish oil and selenium and / or radiotherapy.

[0031] Figure 14 :In Figure 9A Expression of apoptosis markers 24 days after tumor cell implantation in mice treated as shown.

[0032] Figure 15A and 15B : Figure 15A Results are shown of studies characterizing PDL-1 gene expression in primary and metastatic tumor sites in animal models of human disease treated with supplements containing fish oil and selenium and / or radiation therapy. Figure 15B Results of studies characterizing PD-1 gene expression in primary and metastatic tumor sites in animal models of human disease treated with supplements containing fish oil and selenium and / or radiation therapy are shown.

[0033] Figure 16A and 16B : Figure 16A A test protocol is described in which mice received radiation therapy on days 8, 10, and 12 and were provided with nutritional supplements prior to, at the time of, or at the start of radiation therapy. Mice were sacrificed 21 days after tumor cell implantation. Figure 16B :Derived from Figure 16A Treatment groups for the regimens shown in .

[0034] Figure 17 :pass Figure 16A Micrographs of cross-sections of the intestine of mice treated with the protocol described in .

[0035] Figures 18A to 18F : Figure 18A : The therapeutic effects of radiotherapy, NutraWell supplementation, and combination therapy on VEGF gene expression 21 days after tumor cell injection. Figure 18B: The therapeutic effects of radiotherapy, NutraWell supplementation, and combination treatment on BAX gene expression 21 days after tumor cell injection. Figure 18C : The therapeutic effects of radiotherapy, NutraWell supplementation, and combination therapy on Bcl-2 gene expression 21 days after tumor cell injection. Figure 18D : The therapeutic effects of radiotherapy, NutraWell supplementation, and combination treatment on caspase 3 gene expression 21 days after tumor cell injection. Figure 18E : Effects of treatment with radiation, NutraWell supplements, and combination therapy on Bcl-2 gene expression in the lung (ie, metastases) 21 days after tumor cell injection. Figure 18F : Effects of treatment with radiotherapy, NutraWell supplements, and combination treatment on caspase 3 gene expression in the lung (ie, metastases) 21 days after tumor cell injection.

[0036] Figure 19A and 19B : Figure 19A Depicts a test protocol in which mice received radiation therapy on days 8, 10, and 12 and were provided with nutritional supplements for 7 days prior to tumor cell implantation. Mice were sacrificed on day 24 after tumor cell implantation. Figure 19B :Derived from Figure 19A Treatment groups for the regimens shown in .

[0037] Figure 20A and 20B : Figure 20A :use Figure 19A The scheme shown in Figure 2 shows the effect of nutritional supplementation containing fish oil and selenium on body weight in mice receiving repeated radiotherapy after tumor cell implantation. Figure 20B :use Figure 19A The regimen shown in Figure 2 shows the effects of radiation therapy, nutritional supplements containing fish oil and selenium, and combination therapy on gastrocnemius muscle mass 21 days after tumor cell injection.

[0038] Figure 21A and 21B : Figure 21A Depicted is a test protocol in which mice received radiation therapy on days 8, 10, and 12 and were provided with nutritional supplements 7 days before, on the day of, or 8 days after tumor cell implantation. Mice were sacrificed 21 days after tumor cell implantation. Figure 21B :Derived from Figure 21A Treatment groups for the regimens shown in .

[0039] Figure 22 :use Figure 21AFigure 4. Effect of NutraWell supplementation on body weight in mice receiving repeated radiotherapy following tumor cell implantation.

[0040] Figure 23 :When pressing Figure 21A Changes in tumor volume over time in mice treated with the regimen shown in Figure 5. Repeated radiation therapy combined with NutraWell supplementation.

[0041] Figure 24A and 24B : Figure 24A : In use Figure 21A Figure 4. Treatment effects of repeated radiotherapy in combination with NutraWell supplementation and combination therapy on serum TNF-α 21 days after tumor cell injection in mice treated with the regimen shown in FIG. Figure 24B : In use Figure 21A Mice treated with the regimen shown in , combined with repeated radiotherapy and NutraWell supplementation 21 days after tumor cell injection and the therapeutic effect of the combination treatment on serum IL-6. DETAILED DESCRIPTION

[0042] The following description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication explicitly or implicitly referenced is prior art.

[0043] The subject matter of the present invention provides compositions and methods in which nutritional supplements including fish oil and selenium (e.g., supplements containing fish oil, selenium derived from selenium yeast, and certain vitamins, minerals, amino acids, and sugars, such as "NutraWell") are used in combination with radiotherapy. Surprisingly, the combination therapy of radiation and such supplements provides a significant synergistic effect in reducing tumor size. In addition, the side effects of radiotherapy (e.g., neutropenia, weight loss, loss of muscle mass, damage to gastrointestinal acyl cells, etc.) are reduced and / or alleviated relative to the use of radiotherapy without such supplements. Surprisingly, it has also been found that the expression of genes associated with angiogenesis and apoptosis in tumor cells is regulated using supplements containing fish oil and selenium, with or without the application of radiotherapy. In addition, metastasis can be prevented and the growth and spread of cancer stem cells can be reduced.

[0044] It will be appreciated that the disclosed technology provides a number of advantageous technical effects, including enhancing the effectiveness of current radiation therapy regimens used to treat cancer while mitigating the side effects associated with these approaches.

[0045] The following discussion provides many example embodiments of the subject matter of the present invention. Although each embodiment represents a single combination of elements of the present invention, the subject matter of the present invention is considered to include all possible combinations of the disclosed elements. Thus, if one embodiment includes elements A, B, and C, and a second embodiment includes elements B and D, the subject matter of the present invention is considered to include other remaining combinations of A, B, C, or D, even if not explicitly disclosed.

[0046] In some embodiments, the number of components used to describe and claim certain embodiments of the present invention, characteristics such as concentration, reaction conditions, etc., should be understood as being modified by the term "about" in some cases. Therefore, in some embodiments, the numerical parameters set forth in the written specification and the appended claims are approximate values ​​that can be attempted to vary according to the desired characteristics obtained in a specific embodiment. In some embodiments, the numerical parameters should be interpreted according to the number of significant figures reported and by applying common rounding techniques. Although the numerical ranges and parameters illustrating the wide range of some embodiments of the present invention are approximate values, the numerical values ​​illustrated in the specific examples should be reported as accurately as possible. The numerical values ​​presented in some embodiments of the present invention may contain certain errors that are necessarily due to the standard deviation found in their respective test measurements.

[0047] In one embodiment of the present inventive concept, a nutritional supplement composition ("Nutrawell") as shown in Table 1 is provided to improve radiotherapy results.

[0048] Table 1

[0049]

[0050]

[0051]

[0052]

[0053] The compositions shown in Table 1 include components with various physiological and biochemical effects, including anti-inflammatory activity, reduction in blood sugar levels, reduction in cholesterol and anti-tumor activity. Other components provide the supplementation of essential vitamins, minerals and amino acids under elevated levels. When consumed, other components (such as enzymes, lecithin) are used to help digest and absorb the components in the compositions. The combination of these complementary activities provides a synergistic effect that exceeds the simple cumulative effect of a single component. It should be appreciated that the compositions shown in Table 1 also include certain edible spices (such as brown sugar, honey, vanilla flavoring and masking agents) for improving palatability and acceptability. Certain components (such as honey, brown sugar, milk, rice protein, casein) can provide both local flavor and heat. The inventors have found that the combination of above-mentioned edible spices effectively provides the consumption of the nutritional supplement that meets an effective dose. In some embodiments, this edible spice can be excluded without adversely affecting the effectiveness of the nutritional supplement.

[0054] The components shown in Table 1 can be provided as a single formulation (e.g., pill, tablet, capsule, powder, liquid, suspension, etc.), or can be separated into different formulations (e.g., pill, tablet, capsule, powder, liquid, suspension, or a combination thereof). The amount shown in Table 1 is exemplary, and represents the typical daily dose provided to an adult with normal height and normal health in other respects. These amounts can be adjusted to account for differences such as weight, sex, medical conditions, etc. For example, a relatively small patient weighing 40 kilograms or less may benefit from a dosage below the lower limit (low end) or lower limit of the provided range, while a relatively large patient weighing 100 kilograms or more may need to provide a dosage at the upper limit (high end) (or higher) of the scope. In some embodiments, such a daily dose can be distributed as multiple doses throughout the day. In some such embodiments, the composition of each such distributed dose can be identical. In other embodiments, the composition of this distributed dose can be different, as long as the sum of these doses provides the required supplement.

[0055] In an exemplary embodiment, human tumor cells are treated (after transplantation into nude mice) with either 1 gram of nutritional supplements per day, radiation therapy, or 1 gram of nutritional supplements per day and chemotherapy. Mice are weighed during treatment to characterize side effects, such as nausea and loss of appetite. After several weeks, mice are sacrificed and tumors are characterized. Tumor volumes are determined, and the effects of treatment on organ and muscle volume are determined. The degree of neutropenia is also characterized. Table 2 lists a typical regimen.

[0056] Table 2

[0057]

[0058] Weight / weight loss

[0059] exist Figure 1 A typical treatment regimen is schematically depicted in . Some subjects received treatment with a supplement containing fish oil and selenium before tumor cell implantation (PTN, PTRN), and some of these received radiotherapy (PTRN). Other subjects started treatment with this supplement at the time of tumor cell implantation (TN, TRN), and some of these received radiotherapy (TRN). Figure 2A and 2B The results of a weight study compared to control subjects (C) and subjects implanted with tumor cells and not otherwise treated (T) are shown in Figure 3. As shown, mice that received radiation therapy and a supplement containing fish oil and selenium gained weight at a significantly greater rate than those that received radiation therapy alone, indicating a reduction in the side effects typically associated with this treatment modality. Pretreatment with a nutritional supplement containing fish oil and selenium had a particularly pronounced effect.

[0060] Tumor size

[0061] Figures 3A to 3D demonstrated a typical synergistic effect on tumor volume and weight of co-treatment with radiotherapy and supplements containing fish oil and selenium for similar treatment groups in a mouse model of human cancer. Figure 3A Shown are the effects of various treatment regimens on tumor volume over the course of 3 weeks. Figure 3B A zoomed-in view of the effects over the first 11 days is provided. As shown, treatment with the supplement containing fish oil and selenium alone reduced tumor volume by approximately 60%. Treatment with radiation alone provided a similar reduction in tumor volume. Therefore, in the absence of synergy, one would expect to reduce tumor volume to approximately 25% of the volume of the untreated tumor. Surprisingly, a greater than 90% reduction in tumor volume, to approximately 7% of the untreated tumor, was observed—indicating a significant synergistic effect. Figure 3C It was shown that similar effects were found when tumor weight was characterized. Figure 3D Photographs of representative examples of tumors excised from experimental animals after treatment are shown. The effects of synergistic therapy with a nutritional supplement comprising fish oil and selenium and radiation therapy are readily apparent upon visual inspection. Thus, it is apparent that synergistic therapy with a nutritional supplement comprising fish oil and selenium and radiation therapy can provide a synergistic effect in reducing tumor volume and / or mass.

[0062] muscle wasting

[0063] The side effects of radiotherapy are not limited to loss of appetite and weight loss and can include damage to internal organs, loss of muscle mass, anemia, neutropenia, decreased renal function, etc. To determine the protective effect of synergistic therapy with supplements containing fish oil and selenium, this side effect was also characterized with respect to muscle mass and organ weights of treated mice after treatment. The results are shown in Figures 4A to 4E middle. Figure 4A and 4B The effects of treatment with a supplement containing fish oil and selenium, radiotherapy, and synergistic therapy using this supplement (with and without pre-treatment with the supplement) and radiotherapy on the weight of the gastrocnemius and soleus muscles (respectively) are shown, with the test groups designated as described above. As shown, synergistic therapy with this supplement and radiotherapy can provide almost complete preservation of muscle weight.

[0064] transfer

[0065] Figure 4C 、 4D and 4E show the lung weight including any metastatic tumors as described above ( Figure 4C ), liver weight including any metastatic tumors ( Figure 4D ) and spleen weight including any metastatic tumors ( Figure 4E ) treatment effect. As shown, the lack of treatment resulted in weight gain, at least in part due to the presence of metastatic tumors. This gain was not completely alleviated by radiotherapy (TR) alone; however, subjects treated with a supplement containing fish oil and selenium had similar organ weights to control subjects who received co-therapy with and without radiotherapy.

[0066] Similarly, the presence of metastatic sites in the lungs of animal models of human disease treated with nutritional supplements containing fish oil and selenium and / or radiotherapy was quantified. Figure 4F Typical metastatic sites are shown in the photographs in the left panel and listed in the bar graph in the right panel. As shown, treatment with either the supplement (TN) or radiotherapy (TR) reduced the number of metastatic sites to some extent, but treatment with both the supplement and radiotherapy (TRN) resulted in a complete absence of obvious metastatic sites in most subjects.

[0067] Without wishing to be bound by theory, the inventors believe that the effects on metastasis (and tumor size and mass as shown above) may be attributed to the synergistic effect on tumor cell proliferation provided by the use of a nutritional supplement comprising fish oil and selenium in combination with radiation therapy. Figure 4G As shown in the figure, treatment with a supplement containing fish oil and selenium (TN), with radiation therapy (TR), or with both modalities (TRN) inhibited the expression of the proliferation marker Ki-67 in the primary tumor site. Metastatic sites (right panel) showed a related trend, with either nutritional supplementation or radiation therapy alone leading to a modest decrease in the expression of this proliferation marker, but surprisingly showing a synergistic effect in reducing Ki-67 expression in metastatic sites. This suggests that the use of supplements containing fish oil and selenium can reduce tumor metastasis, including metastasis that cannot be prevented by radiation therapy, and can do so in a synergistic manner.

[0068] Neutropenia / anemia

[0069] Anemia and neutropenia are commonly seen in cancer and as side effects of radiation therapy, at least in part due to suppression of bone marrow activity. Figures 5A to 5E The effects of synergistic therapy with supplements containing fish oil and selenium on various blood cell populations were shown in . Figure 5B Nutritional supplementation containing fish oil and selenium, radiotherapy, and synergistic therapy with this supplement and radiotherapy have been shown to have an effect on Figure 5B White blood cells and Figure 5C This suggests that nutritional supplementation containing fish oil and selenium may be useful in ameliorating the suppression of erythrocytes, granulocytes, leukocytes, and / or lymphocytes that results from the presence of tumors and from radiation therapy used in the treatment of such tumors.

[0070] like Figure 5A As shown in Figure 2, platelets were found to be elevated in untreated tumor-bearing (T) animals. Platelet concentrations could be reduced by treatment with a supplement comprising fish oil and selenium or by radiotherapy, but the reduction was most pronounced in animals receiving a synergistic combination of such supplements and radiotherapy. This effect was particularly pronounced in subjects who received pretreatment with a supplement comprising fish oil and selenium, which reduced platelet concentrations to values ​​found in control animals. Granulocyte concentrations were similarly elevated in untreated tumor-bearing animals (e.g., Figure 5D ) and was further elevated in animals that received only radiation therapy. It has been found that synergistic therapy with nutritional supplements containing fish oil and selenium and radiation therapy can reduce this effect. Figure 5E As shown in , tumor-bearing animals exhibited an elevated neutrophil / lymphocyte ratio (NLR) relative to control animals, with the neutrophil / lymphocyte ratio being even higher in similar subjects who received only radiotherapy. Treatment with a supplement comprising fish oil and selenium was found to be effective in shifting this ratio toward more normal values, both as a monotherapy and as part of a synergistic therapy in conjunction with radiotherapy. This suggests that nutritional supplements comprising fish oil and selenium can be used to reduce platelet concentrations, granulocyte concentrations, and / or neutrophil / lymphocyte ratios that are elevated due to the presence of tumors and radiotherapy used to treat such tumors.

[0071] serum proteins

[0072] exist Figure 6A and 6B The effects of synergistic therapy combining supplements containing fish oil and selenium on various serum biochemical markers that provide information on renal function, liver function and / or nutritional status are shown in . Figure 6AThe effects of a nutritional supplement comprising fish oil and selenium, radiotherapy, and a synergistic therapy of such a supplement and radiotherapy on an animal tumor model are shown. As shown, serum albumin concentration (an indicator of nutritional status) is reduced in untreated tumor-bearing animals and is only slightly improved by radiotherapy alone. Treatment with a nutritional supplement comprising fish oil and selenium, whether as a monotherapy or in combination with radiotherapy, can increase serum albumin concentration, particularly when such a supplement is provided as a pretreatment. This is increased by combining a synergistic therapy with a nutritional supplement comprising fish oil and selenium. This suggests that treatment with a nutritional supplement comprising fish oil and selenium, whether as a monotherapy or in combination with radiotherapy, can improve the nutritional status of tumor-bearing subjects.

[0073] Figure 6B Results from a similar study were presented, in which serum creatinine (a measure of kidney function) was characterized. As shown, untreated tumor-bearing subjects showed elevated creatinine concentrations, indicating kidney damage. This was slightly improved by radiotherapy alone. However, a synergistic therapy of a nutritional supplement containing fish oil and selenium and radiotherapy showed a synergistic effect in reducing serum creatinine concentrations, particularly when the nutritional supplement was provided as a pretreatment.

[0074] cytokines

[0075] The growth and spread of tumors are associated with inflammation, as is the use of radiation therapy. Surprisingly, the inventors found that a synergistic therapy combining supplements containing fish oil and selenium effectively reduced the concentration of pro-inflammatory cytokines, suggesting that this combined treatment could be effective in reducing inflammation associated with tumors and radiation therapy of tumors. Figure 7A and 7B The effects of NutraWell supplementation on serum concentrations of pro-inflammatory cytokines were shown in . Figure 7A The concentration of IL-6 in control animals, untreated tumor-bearing animals and tumor-bearing animals treated with nutritional supplements, radiotherapy or a combination of such supplements and radiotherapy comprising fish oil and selenium has been shown. As shown, untreated tumor-bearing animals show that serum IL-6 concentration increases greatly. Radiotherapy can only slightly reduce it. It has been found that the use of nutritional supplements comprising fish oil and selenium can reduce the serum IL-6 concentration in tumor-bearing animals, whether as a monotherapy or in conjunction with the synergistic therapy of radiotherapy. Figure 7B Results from similar studies for IL-1β were presented. The results for IL-1β were similar to those for IL-6. This suggests that nutritional supplementation containing fish oil and selenium, either as monotherapy or as a synergistic therapy combined with radiotherapy, can effectively reduce serum concentrations of inflammation-related cytokines in tumor-bearing subjects. The inventors believe that this reduction is accompanied by a reduction in inflammation in these animals.

[0076] Tumor gene expression

[0077] Surprisingly, the inventors have also discovered that treatment with a nutritional supplement comprising fish oil and selenium can alter gene expression in tumor cells in vivo and can provide a synergistic effect on this alteration in gene expression caused by radiotherapy. In some embodiments, the gene is associated with a cytokine and / or is associated with apoptosis. Examples of the effects of radiotherapy, treatment with a nutritional supplement comprising fish oil and selenium, and synergistic therapy in combination with radiotherapy on gene expression in tumor cells implanted in vivo are shown in Figures 8A to 8G. Figure 8A The results of this treatment on VEGF expression in tumor cells are shown. As shown, monotherapy with a nutritional supplement comprising fish oil and selenium and radiotherapy reduced VEGF expression. Co-therapy with a nutritional supplement comprising fish oil and selenium and radiotherapy provided significantly reduced VEGF expression.

[0078] Figure 8B and 8D The effects of treatment with a nutritional supplement comprising fish oil and selenium, radiotherapy, and a synergistic therapy with such a supplement and radiotherapy on tumor BAX expression and metastatic (lung) tumor BAX expression are shown separately. BAX is considered a marker of apoptosis. As shown, BAX expression in untreated tumors is low and is not affected by radiotherapy. Monotherapy with a nutritional supplement comprising fish oil and selenium results in a dramatic increase in BAX expression, and when used as a synergistic therapy in combination with radiotherapy (particularly when the supplement is provided as a pretreatment), BAX expression increases. Figure 8C and 8E As shown in the study, Bcl-2 expression was found to be elevated in tumor cells and metastatic (lung) tumor cells (respectively) and was reduced by treatment with a nutritional supplement containing fish oil and selenium or radiotherapy as monotherapy. When this supplement and radiotherapy were used as synergistic therapies, a greater reduction in Bcl-2 expression was found. Expression of caspase 3, which is associated with apoptosis, was elevated in tumors treated with radiotherapy or a nutritional supplement containing fish oil and selenium alone. Figure 8D ), and synergistic therapy with this supplement during radiotherapy also increased caspase 3 expression (especially when the supplement was given as pretreatment). Figure 8F As shown in , the expression of caspase 3 is reduced in metastatic (lung) tumors relative to lung tissue from control subjects. As shown, the expression of caspase 3 in such tumors is increased by radiation therapy or treatment with nutritional supplements containing fish oil and selenium and co-therapy.

[0079] Repeated radiotherapy

[0080] A similar study was conducted using a modified treatment regimen. Figure 9A and 9B An improved treatment regimen utilizing multiple rounds of radiation therapy, which is typical of human radiation therapy, is shown in FIG.

[0081] 10A to 10C show the use of Figure 9A Results of serum albumin and blood cell characterization studies following treatment with NutraWell, radiotherapy, and a combination of nutritional supplementation containing fish oil and selenium and radiotherapy, according to the regimen shown in . Figure 10A As shown in , serum albumin concentrations in tumor-bearing animals decreased, especially at later time points. This could be improved by using nutritional supplements containing fish oil and selenium or radiotherapy as monotherapy as well as by co-therapy. Figure 10B As shown in the study, tumor-bearing animals displayed suppressed lymphocyte counts relative to control subjects, particularly at later time points. This was only slightly improved by radiotherapy alone, but treatment with a nutritional supplement containing fish oil and selenium, either as monotherapy or in combination with radiotherapy, effectively increased lymphocyte concentrations, particularly at later time points. Figure 10C Results from a similar study were presented in which the neutrophil / lymphocyte ratio (NLR) was characterized. As shown, tumor-bearing animals showed a sharp increase in this value at later time points. This can be reduced by treatment with nutritional supplements containing fish oil and selenium, radiotherapy, and a combination of such supplements and radiotherapy.

[0082] Surprisingly, using Figure 9A The regimens shown in , with nutritional supplements containing fish oil and selenium, radiotherapy, and combined treatment with such supplements and radiotherapy also had an effect on the expression of tumor cell markers and tumor cell metastasis. In the following studies, the tumor cells selected for implantation were derived from lung tumors and had a strong tendency to metastasize to the lungs from the implantation site. Figures 11A to 11D Shows the Figure 9A Results of immunocytochemistry studies of different tissues from mice treated with the regimens shown in Figure 2. Notably, NutraWell supplementation alone reduced or eliminated metastases.

[0083] like Figure 11A As shown in Figure 3, VEGF expression is evident in tumors of untreated mice (left panel, FITC staining). As with the combined therapy, treatment with nutritional supplements containing fish oil and selenium or radiation therapy alone significantly reduced VEGF expression. Figure 11A Numerical results are provided in the right figure of . Figure 11B Similar results were found in tumors that had metastasized to the lungs, as shown in Figure 2. As shown, VEGF expression was evident in untreated metastatic tumors (left panel, FITC staining) and was dramatically decreased in animals that received nutritional supplements containing fish oil and selenium, radiation therapy, or both. Figure 11B The numerical results of these studies are provided in the right figure of .

[0084] Similar results were found in EGFR expression, where overexpression of EGFR is associated with tumors. Figure 11C As shown in Figure 3, elevated EGFR expression (left panel, FITC staining) was evident in untreated tumors and was reduced in subjects treated with nutritional supplements containing fish oil and selenium, radiotherapy, or a combination of such supplements and radiotherapy. Figure 11C The numerical results are shown in the right figure of . Figure 11D As shown in, for the EGFR expression in the tumour that has transferred to lung, similar result has been found.As shown, the EGFR expression (left figure, FITC dyeing) that rises is obvious in untreated metastasis site, and with the nutritional supplement or radiotherapy that comprises fish oil and selenium as monotherapy, and use this supplement and the experimenter of radiotherapeutic synergistic therapy process to reduce.This shows that use the nutritional supplement, radiotherapy and / or this supplement and radiotherapy combination process that comprise fish oil and selenium can effectively strengthen the treatment plan for EGFR in the individuality that accepts cancer treatment.

[0085] Surprisingly, using Figure 9A The regimen shown in [ 1 ], treatment with a nutritional supplement containing fish oil and selenium, radiotherapy, and a combination of these supplements and radiotherapy also have an effect on cancer stem cells. These stem cells are implicated in metastasis and progression resistant to various cancer therapies. In the following studies, the tumor cells selected for implantation were derived from lung tumors and have a strong tendency to metastasize to the lungs from the implantation site. Figure 12 Shows the Figure 9A Results of immunocytochemical studies of different tissues from mice treated with the regimens shown in . Figure 12 The left panel shows the results of staining (FITC staining) for the stem cell marker CD31 in tumor cells. The right panel shows similar results for lung metastatic cells. Untreated subjects showed a large number of cells with elevated CD31 expression. Surprisingly, treatment with a nutritional supplement containing fish oil and selenium, in the absence of radiation therapy, reduced or eliminated the occurrence of cancer stem cells at the tumor implantation site and lung metastasis sites.

[0086] use Figure 9A The regimens shown in [ 1 ], nutritional supplementation with fish oil and selenium, radiation therapy, and combined NutraWell supplementation and radiation therapy treatment also had an effect on the hypoxia typically found within or between tumor cells. HIF1-α is a marker associated with hypoxia. In the following studies, the tumor cells selected for implantation were derived from lung tumors and had a strong tendency to metastasize from the implantation site to the lungs. Figure 13A Shows the Figure 9AResults of immunocytochemical studies of different tissues from mice treated with the regimen shown in Figure 1. The left panel shows the results of immunocytochemical staining (FITC staining) for HIF1-α in tumor cells, while the right panel shows the results of similar staining of lung metastases. Untreated subjects showed a large number of cells with elevated HIF1-α levels. Surprisingly, treatment with a nutritional supplement containing fish oil and selenium in the absence of radiation reduced or eliminated the appearance of this hypoxia marker at the tumor implantation site and lung metastases. Figure 13B Typical numerical results from similar studies are shown, wherein gene expression in tumor samples is characterized. As shown, as with radiotherapy (TR), treatment with a supplement comprising fish oil and selenium (TN) and pretreatment with a nutritional supplement comprising fish oil and selenium (PTN) for one week can both reduce tumor expression of HIF1-α. When radiotherapy and this nutritional supplement are used in combination (PTRN, TRN), tumor HIF1-α expression is sharply reduced, particularly in pretreated subjects (PTRN) (which shows synergy). It is clear that nutritional supplements comprising fish oil and selenium can be used in combination with radiotherapy to reduce HIF1-α protein content and gene expression in the tumor site, which may make them more susceptible to the damage of hypoxic conditions.

[0087] Using the protocol shown in 9A, nutritional supplementation containing fish oil and selenium, radiation therapy, and combined supplementation and radiation therapy also had an effect on the apoptotic activity of tumor cells. Figure 14 The results of qPCR studies on the expression of various apoptosis markers (Bax, Bcl-2, and caspase 3) on day 24 from tumor cell implantation in mice are shown. As shown, the Bax / Bcl-2 expression ratio is low in tumor cells and is only slightly improved by radiotherapy. Surprisingly, treatment with a nutritional supplement containing only fish oil and selenium provides a significant increase in this ratio. In addition, a significant synergistic effect was found in the synergistic therapy of this supplement and radiotherapy. Caspase 3 expression was actually suppressed by radiotherapy alone, but was dramatically increased by treatment with a nutritional supplement containing fish oil and selenium, both as a monotherapy and as a synergistic therapy in combination with radiotherapy.

[0088] It has also been found that in animal models of human disease, nutritional supplementation containing fish oil and selenium combined with radiation therapy can modulate the expression of PDL-1 and PD-1 at primary and metastatic (lung) tumor sites. Figure 15AAs shown in the figure, both primary (left) and metastatic (right) tumor cells showed high levels of PDL-1 expression (T). Surprisingly, treatment with a nutritional supplement comprising fish oil and selenium significantly reduced these levels when pre-treated (PTN) and treated at the time of implantation (TN). Therefore, the inventors believe that this supplement can be used to enhance immunotherapy methods for the treatment of cancer. Treatment with radiotherapy (TR) also showed that PD-L1 gene expression was reduced at both the primary site and the metastatic site, suggesting that radiotherapy alone can be used to enhance immunotherapy methods for the treatment of cancer. Whether the supplement is provided before implantation (PTRN) or at the time of implantation (TRN), synergistic therapy with this nutritional supplement and radiotherapy can effectively reduce PDL-1 expression in the primary tumor site and the metastatic site. Surprisingly, the effect was more significant at the primary tumor site than at the metastatic site. This suggests that synergistic therapy combining nutritional supplements including fish oil and selenium with radiotherapy may make tumor cells at both primary and metastatic sites more susceptible to the patient's immune system and / or may enhance the effectiveness of immunotherapy approaches for cancer treatment.

[0089] Similar studies have been conducted on PD-1 expression. Figure 15B As shown in the , PD-1 expression was decreased in primary tumor cells treated with radiation therapy (TR) both before implantation (PTN) and at the time of implantation (TN), whereas it was increased in primary tumor cells in animals treated with supplements containing fish oil and selenium (left panel). Synergistic therapy with nutritional supplements and radiation therapy may provide different results depending on whether the supplements are provided before implantation (PTRN) or at the time of implantation (TRN). As shown in Figure 15B As shown in the right figure in , tumor cells at the metastatic site showed reduced PD-1 expression relative to samples obtained from control animals. Surprisingly, radiotherapy resulted in an increase in PD-1 expression at the metastatic site, both before and at the time of implantation (right figure), as did treatment with a supplement comprising fish oil and selenium. The synergistic therapy of radiotherapy and this supplement can provide higher levels of PD-1 expression than those observed with monotherapy. It is apparent that treatment with a nutritional supplement comprising fish oil and selenium, particularly in combination with radiotherapy, can move tumor-bearing subjects, particularly PD-1 expression observed at the metastatic site, away from reduced expression levels.

[0090] Supplemental and repeat radiotherapy before implantation, before radiotherapy, and at the start of radiotherapy.

[0091] exist Figure 16A and 16B Another treatment regimen that combines nutritional supplements containing fish oil and selenium with pretreatment with multiple rounds of radiation therapy, typical of clinical practice, is shown in In this regimen, treatment with the supplements is initiated before tumor cell implantation, at the time of tumor cell implantation, and at the start of radiation therapy.

[0092] Loss of intestinal absorption and resulting malnutrition are well-known side effects of radiotherapy, especially repeated radiotherapy. Figure 17 Shown and show the photomicrograph of the treatment effect of the nutritional supplement that comprises fish oil and selenium to visceral cellular structure during radiotherapy.Shown the cellular structure of the internal organs of untreated control subject (upper left figure), the cellular structure of the internal organs of the experimenter of the implanted tumor of many rounds of radiotherapy treatment (upper right figure), with the nutritional supplement pretreatment that comprises fish oil and selenium and subsequently with the cellular structure of the internal organs of the experimenter of the implanted tumor of radiotherapy treatment (lower left figure), and when starting radiotherapy, with the cellular structure of the internal organs of the experimenter of the implanted tumor of fish oil and selenium treatment (lower right figure).As shown, can maintain intestinal brush border during radiotherapy with the nutritional supplement processing that comprises fish oil and selenium, and pretreatment obviously strengthens intestinal brush border.This shows to process with this supplement, particularly pretreatment, can effectively solve the side effect of radiotherapy.

[0093] This treatment regimen has been found to modify the expression of certain genes in tumor cells in vivo. Figure 16A Results of qPCR studies on gene expression (e.g., angiogenesis factor-related, apoptosis-related, etc.) in tumor cells from mice treated with the regimen shown in FIG. Figure 18A As shown in Figure 2, treatment with a nutritional supplement comprising fish oil and selenium reduced VEGF expression in tumor cells, as did repeated radiotherapy. Combination therapy with this supplement and radiotherapy provided an enhanced reduction in VEGF expression, particularly when the nutritional supplement comprising fish oil and selenium was provided before the start of radiotherapy.

[0094] Figure 18B It was shown that BAX expression in tumor cells was increased by treatment with a nutritional supplement containing fish oil and selenium and was relatively unaffected by repeated radiotherapy alone. Pretreatment with this supplement in combination with repeated radiotherapy also increased BAX expression. Figure 18D Similar results were found for the expression of another apoptosis-related gene (caspase 3) as shown in . Figure 18F Results of a study on caspase 3 expression in metastatic (lung) tumors are shown in As shown, caspase 3 expression in such metastatic tumors can be increased by treatment with a nutritional supplement containing fish oil and selenium and by repeated radiotherapy, and when the supplement is given as a co-therapy before the start of radiotherapy, caspase 3 expression is similar to that in control cells.

[0095] The expression of Bcl-2, another apoptosis-related gene, was reduced in tumor cells by treatment with nutritional supplements containing fish oil and selenium and by repeated radiation therapy alone (see Figure 18CThis reduction in Bcl-2 expression was more pronounced with the combined use of supplementation and repeated radiotherapy, particularly when supplementation was given before the start of radiotherapy. Figure 18E Results from a similar study conducted on metastatic (lung) tumors are shown. As shown, the reduction in Bcl-2 expression in the combined therapy of nutritional supplementation comprising fish oil and selenium and repeated radiotherapy was reduced in a synergistic manner, particularly when the supplementation was provided before the start of radiotherapy.

[0096] Supplemental and repeat radiotherapy before implantation

[0097] Figure 19A and 19B The treatment regimen and the associated test groups are shown (respectively), wherein nutritional supplements containing fish oil and selenium were provided 7 days prior to tumor cell implantation, and radiotherapy was performed on days 8, 10, and 12 after implantation. Mice were sacrificed on day 24 after implantation.

[0098] A well-known side effect of both cancer and repeated radiation therapy is weight loss. This can be due to both disease-related emaciation and the side effects of radiation therapy. Figure 20A and 20B The effects of treatment with a nutritional supplement containing fish oil and selenium on weight loss and muscle mass following repeated radiotherapy are shown in FIG. It should be recognized that body weight was characterized after removal of the tumor mass. Figure 20A As shown in the study, when the tumor mass was removed, a significant reduction in the remaining body weight was evident relative to control subjects. This was improved by treatment with a nutritional supplement containing fish oil and selenium and by radiotherapy as a monotherapy. Surprisingly, when this supplement was combined with repeated radiotherapy, weight gain exceeded that of control subjects. Figure 20B The weight of the gastrocnemius muscle in each test group is shown. In untreated tumor-bearing animals, the loss of muscle mass is obvious. It can be slightly improved by repeated radiotherapy alone. Surprisingly, muscle mass can be significantly retained by treating with a nutritional supplement comprising fish oil and selenium. Relative to untreated tumor-bearing subjects, a synergistic improvement in muscle mass is provided by a synergistic therapy of a nutritional supplement comprising fish oil and selenium with repeated radiotherapy. It is obvious that pretreatment with a nutritional supplement comprising fish oil and selenium can effectively reverse the weight loss (relative to control subjects) and the loss of muscle mass caused by the presence of tumors and repeated radiotherapy.

[0099] Preimplantation embryo supplementation and repeat radiotherapy

[0100] exist Figure 21A and 21B Another treatment regimen for the relevant treatment group is shown in Figure 16A and 16BThe protocol is similar to that shown in . In this protocol, radiotherapy was provided on days 8, 10, and 12 after tumor cell implantation. Treatment with a nutritional supplement containing fish oil and selenium was provided 7 days before implantation, on the day of implantation, or at the start of radiotherapy. Mice were sacrificed on day 21 after tumor cell implantation.

[0101] A well-known side effect of cancer and radiation therapy is weight loss. Figure 22 In the present invention, the effects of a nutritional supplement comprising fish oil and selenium on weight loss and muscle mass loss when provided before and simultaneously with initial repeated radiotherapy are shown. It should be understood that body weight was characterized after removal of the tumor mass. As shown, relative to control subjects, the weight gain (after excision of the tumor) of untreated tumor-bearing subjects over time was significantly reduced. Similar losses were recorded when only repeated radiotherapy was used. Surprisingly, whether as a monotherapy or when used in combination with repeated radiotherapy, a nutritional supplement comprising fish oil and selenium was used to significantly improve weight gain over time. This was particularly evident when this supplement was used as a pretreatment.

[0102] The inventors also discovered that Figure 23 As shown in Figure 2, pretreatment with a nutritional supplement comprising fish oil and selenium enhanced the reduction in tumor volume observed in repeated radiotherapy. As shown, both repeated radiotherapy and treatment with a nutritional supplement comprising fish oil and selenium had a modulating effect on reducing tumor volume relative to tumors in untreated tumor-bearing animals. Surprisingly, when repeated radiotherapy and a nutritional supplement comprising fish oil and selenium were used as synergistic therapies, tumor volume barely changed over time, indicating a synergistic effect.

[0103] The presence of a tumor and repeated radiotherapy may also lead to the development of inflammation, characterized by the presence of proinflammatory cytokines in the serum. Figure 24A and 24B showed that administration of a nutritional supplement containing fish oil and selenium and repeated radiotherapy had an effect on Figure 21A Effects of the regimens shown in [ 001 ] on the concentrations of proinflammatory cytokines in mice treated with [ 001 ] . Figure 24A Serum TNF-α values ​​are shown. Clearly, untreated tumor-bearing animals exhibit highly elevated TNF-α concentrations, which can be reduced to some extent by repeated radiotherapy. Treatment with a nutritional supplement containing fish oil and selenium also results in a reduction in serum TNF-α, particularly when this supplement is used in conjunction with repeated radiotherapy. Figure 24B Results of a similar study are shown, in which serum concentrations of IL-6 were characterized, which showed similar results.

[0104] It should be apparent to those skilled in the art that, in addition to those already described, further modifications may be made without departing from the inventive concepts herein. Therefore, the subject matter of the present invention is not limited except in the spirit of the appended claims. Furthermore, when interpreting the specification and claims, all terms should be interpreted in the broadest possible manner consistent with the context. In particular, the terms "comprise" and "comprising" should be interpreted as referring to elements, components or steps in a non-exclusive manner, indicating that the referenced elements, components or steps may be present, utilized or combined, as well as other elements, components or steps not explicitly referenced. Where at least one of A, B, C ... and N is cited in the specification and claims, the text should be interpreted as requiring only one element in the group, rather than A plus N or B plus N, etc.

Claims

1. A method for treating a tumor, comprising: Apply radiation therapy protocols to patients who need treatment; and The patient is provided with a nutritional supplement comprising fish oil and selenium in an amount that provides a synergistic effect in reducing tumor volume or weight.

2. The method of claim 1, wherein the nutritional supplement is provided to the patient prior to commencing radiation therapy.

3. The method of claim 1 or 2, wherein the nutritional supplement is formulated such that the various components of the supplement are provided in the amounts as described in Table 1.

4. The method of claim 1 or 2, wherein the supplement comprises three or more components as described in Table 1.

5. A method for reducing the side effects of radiotherapy, comprising: Apply radiation therapy protocols to patients who need treatment; and The patient is provided with a nutritional supplement comprising fish oil and selenium in an amount to reduce the side effects of the radiation therapy regimen.

6. The method of claim 5, wherein the nutritional supplement is provided to the patient prior to commencing radiation therapy.

7. The method of claim 5 or 6, wherein the nutritional supplement is formulated such that the various components of the supplement are provided in the amounts as described in Table 1.

8. The method of claim 5 or 6, wherein the supplement comprises three or more components as described in Table 1.

9. A method of regulating gene expression in a tumor, comprising: A nutritional supplement is provided to a tumor or an animal having a tumor, wherein the nutritional supplement comprises fish oil and selenium, and wherein the nutritional supplement is provided in an amount that modulates gene expression in the tumor.

10. The method of claim 9, wherein the nutritional supplement is provided to the patient prior to commencing radiation therapy.

Images