JP2014513698A - Use of arsenic for cancer treatment protection - Google Patents

Abstract

  Methods for inhibiting, preventing or reducing damage to non-cancerous cells in a human subject during chemotherapeutic or radiation treatment of cancer cells in a human subject include radiation or one or more chemotherapeutic agents Prior to treatment includes administering to the human subject a therapeutically effective amount of arsenic and / or one or more arsenic compounds.

Description

[Statement concerning research or development supported by the federal government]
This invention was made with government support under grant number CA085679 awarded by the National Institutes of Health. The government has certain rights in the invention.

[Field of the Invention]
The present invention relates generally to the treatment of cancer. More specifically, the present invention is directed to ameliorating the side effects caused by chemical cancer therapy and radiation cancer therapy.

  Cancer is the leading cause of death in animals and humans. Over the last decade, the combination of chemotherapy and radiation therapy in addition to surgery has become the standard approach for the treatment of cancer patients in curative and palliative settings. Radiation and chemotherapy are successful methods of cancer treatment, but they do not distinguish cancerous cells from normal cells. Thus, in the process of killing cancer cells, radiation or chemotherapeutic agents also damage normal tissues, resulting in systemic toxicity and adverse side effects, which often pose a significant threat to cancer patients . Adverse side effects also greatly limit the maximum dose allowed. Despite repeated efforts in the past, efforts to avoid chemotherapy and radiotherapy toxicity have not yielded significant results.

  In one aspect, the invention provides a human subject in need of radiation treatment for the treatment of one or more arsenic compounds with about 1 μg / kg / day to about 125 μg / kg / day protective. The present invention relates to a method for inhibiting, preventing or reducing damage to non-cancerous cells in a human subject during radiation treatment of the cancer cells in a human subject, comprising administering in an appropriate amount. In some embodiments, the protective amount of one or more arsenic compounds is from about 31 μg / kg / day to about 125 μg / kg / day. Radiation is administered to a human subject after administration of one or more arsenic compounds. In some embodiments, arsenic trioxide is administered to the human subject prior to administering radiation treatment to the human subject. In some embodiments, the one or more arsenic compounds are administered to the human subject at least one day prior to administration of radiation treatment to the human subject. In some embodiments, the one or more arsenic compounds are administered to the human subject daily for at least 3 days prior to administration of radiation to the human subject.

  In another aspect, the invention administers one or more arsenic compounds in a protective amount from about 1 μg / kg / day to about 125 μg / kg / day to a human subject in need of chemotherapeutic treatment. A method of inhibiting, preventing or reducing damage to non-cancerous cells in a human subject during chemotherapeutic treatment of cancer cells in a human subject. In some embodiments, the protective amount of one or more arsenic compounds is from about 31 μg / kg / day to about 125 μg / kg / day. The one or more chemotherapeutic agents are administered to the human subject after administration of the one or more arsenic compounds. In some embodiments, arsenic trioxide is administered to the human subject prior to administering radiation treatment to the human subject. In some embodiments, the one or more arsenic compounds are administered to the human subject at least one day prior to administration of the one or more chemotherapeutic agents to the human subject. In some embodiments, the one or more arsenic compounds are administered to the human subject daily for at least 3 days prior to administration of the one or more chemotherapeutic agents to the human subject.

  In another aspect, the present invention relates to cancer in a human subject comprising administering one or more arsenic compounds to a human subject in need of chemotherapeutic treatment or radiation treatment for the treatment of cancer. It relates to methods for inhibiting, preventing or reducing side effects in human subjects during chemotherapeutic or radiation treatment of cells. One or more chemotherapeutic agents or radiation is administered to the human subject after administration of the one or more arsenic compounds. One or more arsenic compounds are administered in an amount sufficient to inhibit, prevent or reduce side effects caused by chemotherapeutic or radiation treatment in a human subject when the chemotherapeutic agent is administered to the human subject. Is done. In some embodiments, the one or more arsenic compounds are administered in a protective amount from about 1 μg / kg / day to about 125 μg / kg / day.

  Side effects that could be inhibited, prevented, or reduced were related to gastrointestinal tract related side effects, low red blood cell count related side effects, low white blood cell count related side effects, low platelet count related side effects, bone marrow cell depletion. Side effects, side effects related to cardiotoxicity, and side effects related to hair loss.

  The advantages of the present invention will become apparent to those of ordinary skill in the art by reference to the accompanying drawings, described below, with the help of the detailed description of the following embodiments.

FIG. 5 shows the effect of arsenic pretreatment on male mice injected with human lung cancer cells under various treatment conditions. FIG. 5 shows the effect of arsenic pretreatment on female mice injected with human lung cancer cells under various treatment conditions. FIG. 5 shows the effect of arsenic pretreatment on female mice injected with human breast cancer cells under various treatment conditions. FIG. 5 shows the effect of arsenic pretreatment on mice injected with human colon cancer cells under various treatment conditions. FIG. 5 shows the effect of arsenic pretreatment on WT mice versus p53 R172P knock-in mice under X-ray irradiation. It is a figure which shows the effect of the arsenic on the white blood cell count and small intestine damage after a radiation treatment.

  While the invention may be susceptible to various modifications and alternative forms, specific embodiments thereof are shown by way of example in the drawings and are herein described in detail. The drawing may not be proportional to the actual size. However, the drawings and detailed description thereof are not intended to limit the invention to the particular forms disclosed, but on the contrary, the intent is the book as defined by the appended claims. It is to be understood that this is intended to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.

The following definitions are provided:
As used herein, the terms “administration”, “administering” and the like, when used in the context of providing a composition to a subject, generally indicate that the compound being administered is administered by the compound. One or more pharmaceutical compositions, “commercially available” (OTC), in combination with a suitable delivery vehicle by any means, so as to achieve one or more of the biological effects intended to be ) Refers to providing a subject with a composition, or nutritional supplement composition. By way of non-limiting example, the composition can be administered by parenteral, subcutaneous, intravenous, intracoronal, rectal, intramuscular, intraperitoneal, transdermal, or buccal delivery routes. Alternatively or simultaneously, administration may be by the oral route. The dose administered depends on the age, health, weight, and / or disease state of the recipient, if any, the type of simultaneous treatment, the frequency of treatment, and / or the nature of the effect desired. The dose of pharmacologically active compound administered depends on the age, health, weight and / or disease state of the recipient, if any, the nature of the simultaneous treatment, the frequency of treatment, and / or the desired biological effect. And depends on several factors such as size.

  As used herein, the term “cancer” is defined as an uncontrolled growth or proliferation of cells, such as a tumor. In certain embodiments, the tumor results in local invasion and metastasis.

  As used herein, the term “chemotherapeutic agent” is defined as a drug, toxin, compound, composition, or biological entity used as a treatment for cancer.

  As used herein, the term “cytotoxin” is defined as a drug, toxin, compound, composition, or biological entity used to kill cells. In an embodiment, the cell is a cancer cell.

  As used herein, the term “DNA damaging agent” is a drug, toxin, compound, composition, or biological entity that damages a nucleic acid. The damage may be of any kind to the nucleic acid, for example, to break one or both strands of the DNA double helix molecule or to cause one or more nucleotide mutations .

  As used herein, the term “drug” is defined as a medicament or drug used for the therapeutic treatment of a medical condition or disease. The drug may be used in combination with another drug or another type of therapy, and in embodiments is effective in treating cancer.

  As used herein, the term “pharmaceutically or pharmacologically acceptable” results in an adverse, allergic or other inconvenient response when administered to an animal or human. Refers to no molecular entity and composition.

  As used herein, the term “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like.

  As used herein, terms such as “pharmaceutical composition”, “pharmaceutical formulation”, “pharmaceutical preparation” generally refer to a defined dose of one or more pharmacologically active compounds. Refers to a formulation adapted for delivery to a cell, group of cells, organ or tissue, animal or human. Methods for incorporating pharmacologically active compounds into pharmaceutical preparations are well known in the art. The determination of an appropriate defined dose of a pharmacologically active compound to be included in a pharmaceutical composition to achieve a desired biological result is within the skill level of those skilled in the art. The pharmaceutical composition may be provided as a sustained or sustained release formulation. Such a formulation can release a large amount of the compound from the formulation at a desired point in time, or can ensure that a relatively constant amount of compound present in a dose is released over a predetermined period of time. . Terms such as “sustained release”, “controlled release”, or “sustained release” are widely used in the pharmaceutical field and are easily understood by those skilled in the art. The pharmaceutical preparation can be prepared as a solid, semi-solid, gel, hydrogel, liquid, solution, suspension, emulsion, aerosol, powder, or combinations thereof. One or more carriers, preservatives, flavoring agents, excipients, coatings, stabilizers, binders, solvents, and / or auxiliaries that are typically pharmacologically inert are included in the pharmaceutical preparation. May be included. It will be readily appreciated by those skilled in the art that pharmaceutically acceptable salts of the compounds are included within the meaning of the term. It will be further understood by those skilled in the art that the term also encompasses a pharmaceutical composition containing a mixture of two or more pharmacologically active compounds, such compounds as, for example, combination therapy Be administered.

  As used herein, the term “subject” generally refers to a mammal, particularly a human. In one embodiment, the subject receiving an arsenic-containing compound is one who is scheduled for chemotherapy or radiation therapy. For example, the subject can be a human patient or animal that has been diagnosed with cancer for which chemotherapy or radiation therapy is considered an advantageous treatment.

  As used herein, the term “treating” is defined as the practice of administering a treatment for a medical condition or disease. A treatment need not provide complete healing and is considered effective if at least one symptom is ameliorated or eradicated. Furthermore, treatment need not provide permanent improvement of the disease state or condition, but this is preferred.

  The terms “in need of treatment”, “in need of”, “benefit from such treatment” when used in connection with a subject to whom a pharmacologically active composition is to be administered “Will be” generally refers to a judgment made by an appropriate health care provider that an individual or animal will require or benefit from a particular treatment or medical intervention. Such decisions can be made based on a variety of factors that are within the domain of healthcare provider expertise, but an individual or animal may be ameliorated or treated with certain medical interventions Includes knowledge that, as a result of the condition, is ill, will or will be at risk.

  “Side effects” associated with chemotherapy or radiotherapy include abdominal pain, hyperacidity, reflux of stomach acid, hair loss (hair loss), anemia, loss of appetite, early satiety, joint pain (nodal pain), lethargy, ataxia , Hypernitrogenemia, hepatotoxicity, bronchitis, constipation, cystitis, deep vein thrombosis (DVT), dyspepsia, dyspnea, edema, esophagitis, granulocytopenia, gynecomastia, hematoma, hemorrhagic cystitis, leukocytes Decrease, mucositis, myalgia, myocarditis, nephrotoxicity, neutropenia, pancytopenia, pericarditis, sore throat, stomatitis, thrombocytopenia, dry mouth, skin dryness, flushing, hyperpigmentation, nail changes , Photosensitivity, radiation recall, and rash. In some embodiments, the side effects include side effects due to neutropenia (low white blood cell count). A low white blood cell count can lead to increased infection in the body. In some embodiments, the side effects include side effects due to anemia (low red blood cell count). A low red blood cell count can lead to side effects such as headache and fatigue. In some embodiments, the side effects include those caused by thrombocytopenia (low platelet count). A low platelet count can lead to side effects such as increased contusion formation, punctate bleeding, and bleeding (eg, nose, gums, rectum). Other side effects include side effects on the gastrointestinal tract (eg, nausea, abdominal pain, abdominal cramps, flatulence (gas), excessive stomach acid, acid reflux, loss of appetite, early satiety, etc.). Other side effects include hair loss (hair loss) and skin reactions (eg, dry skin, flushing, hyperpigmentation, nail changes, photosensitivity, radiation recall, and rash).

  The phrases “therapeutically effective amount” and “effective amount” are synonymous unless otherwise indicated and are sufficient to ameliorate the condition, disease or disorder to be treated. Means the amount of the compound of the invention. The determination of other factors related to effective administration of the compounds of the present invention to patients in need of treatment, including therapeutically effective amounts, as well as dosage forms, routes of administration, and frequency of administration, is faced. May depend on the characteristics of the condition being treated, such as the patient and condition being treated, the severity of the condition in the particular patient, the particular compound being used, Specific administration route to be administered, frequency of administration, and specific formulation to be used. Determination of a therapeutically effective treatment plan for a patient is possible within the level of a person in the medical or veterinary field. For clinical use, an effective amount is S. It can be the amount recommended by Food and Drug Administration, or equivalent foreign government agencies. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending upon the mammalian host being treated and the particular mode of administration.

  As used herein, the term “protective amount” is sufficient to reduce, prevent, or otherwise ameliorate harmful side effects of radiation therapy or chemotherapeutic drugs to normal cells. Describes an effective amount of an arsenic-containing compound that is administered to a subject concurrently, separately or sequentially with radiation therapy or one or more chemotherapeutic agents.

  As used herein, the term “tumor cell” is defined as a cell of a malignant mass, such as a tumor or cancer. The cell may be located within the tumor, on the surface of the tumor, or it may be associated with the tumor.

  As used herein, the terms “reducing”, “inhibiting”, and “ameliorating” when used in connection with modulating a pathological condition or disease state are generally Refers to the prevention and / or reduction of at least some of the negative consequences of the disease state. When used in the context of a biochemical event or pathway, the term generally refers to a net reduction in the size or activity of the pathway.

  It should be understood that the present invention is not limited to a particular chemotherapy or radiation therapy, which can of course vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. As used in this specification and the appended claims, the singular forms “a”, “an”, and “the” do not expressly indicate otherwise. Unless otherwise specified, includes singular and plural referents.

  As understood by those skilled in the art, for any and all purposes, particularly with respect to providing written explanations, all ranges disclosed herein are intended to cover all possible subranges and subranges thereof. Includes combinations. Any listed range is sufficient to describe and enable the same range to be decomposed into at least 2 equal parts, 3 equal parts, 4 equal parts, 5 equal parts, 10 equal parts, etc. Can be easily recognized. As a non-limiting example, each range discussed herein can be easily broken down into a lower third, a middle third, a top third, and so on. Also, as will be understood by those skilled in the art, all languages such as “to”, “at least”, “greater than”, “less than”, etc., include the listed numbers and are discussed above. As such, it refers to a range that can later be broken down into partial ranges. Finally, as understood by those skilled in the art, the range includes each individual member. Thus, for example, a group having 1-3 units refers to a group having 1 unit, 2 units, or 3 units. Similarly, a group having 1 to 5 units refers to a group having 1 unit, 2 units, 3 units, 4 units, or 5 units.

  Most types of cancer can be treated with either chemotherapy or radiation therapy. Arsenic is found in breast cancer, ovarian cancer, colorectal cancer, stomach cancer, lung cancer, kidney cancer, bladder cancer, prostate cancer, ureteral cancer, thyroid cancer, pancreatic cancer, cervical cancer, and esophagus. Cancers such as cancer, mesothelioma, head and neck cancer, hepatocellular carcinoma, melanoma, brain cancer, vulvar cancer, testicular cancer, sarcoma, intestinal cancer, skin cancer, leukemia, and lymphoma Useful in reducing side effects associated with chemotherapy or radiation therapy treatments. Various animal models for such cancers are known and can be used to explore the efficacy of arsenic, as well as administration and dosing protocols. In some embodiments, the subject receives a protective amount of arsenic followed by a chemotherapeutic agent that is not an arsenic-containing compound.

  In an embodiment, there is provided a method in which one or more arsenic compounds are administered to a patient together with the administration of a chemotherapeutic agent and / or radiation to the patient to inhibit the side effects of chemotherapy or radiation therapy in the cancer patient. Provided. The arsenic compound or compounds are administered in an amount effective to reduce side effects, but in an amount that is much less than an amount that would induce additional tumor formation. In some embodiments, the one or more arsenic compounds are administered to the patient prior to administration of chemotherapy or radiation therapy. In embodiments, the one or more arsenic compounds are administered substantially simultaneously with or after administration of chemotherapy or radiation therapy to the patient. The arsenic compound or compounds may be administered before, during and after chemotherapy or radiation therapy. Reduced severity of side effects after chemotherapy and radiation therapy enhances the quality of life experienced by patients receiving chemotherapy or radiation therapy. As an indication of such improved quality of life, cancer patients have a stronger appetite, better sleep, higher levels of energy, less pain compared to patients who have not been pretreated with arsenic It is observed to have less gastrointestinal disturbances, less hair loss, reduced incidence of infection, and desirable weight gain.

  Arsenic can be administered at any point near the administration of a treatment (eg, chemotherapy or radiation therapy) that produces the desired protective effect. In one embodiment, arsenic is administered to a subject prior to administration of the treatment, for example, 1-2 days, 1-3 days, 1-4 days, 1-5 days, or 1-10 days before treatment administration. The In some embodiments, arsenic is administered to the subject 1 day, 2 days, 3 days, 4 days, or 5 days before administration of the treatment. In a suitable embodiment, arsenic is administered 3 days prior to administration of treatment. During this period, administration occurs once daily, twice daily, three times daily, four times daily, six times daily, or substantially continuously (eg, by intravenous administration). Also good.

  Typically, an effective amount of an arsenic composition sufficient to achieve a protective effect ranges from about 5 μg per kilogram body weight per day to about 3,500 μg per kilogram body weight per day. . In some embodiments, an effective amount ranges from about 10 μg per kilogram body weight per day to about 1,000 μg per kilogram body weight per day. In other embodiments, the effective amount is from about 5 to about 1,500 μg / kg / day, from about 5 to about 1,000 μg / kg / day, from about 5 to about 850 μg / kg / day, from about 5 to about 500 μg. / Kg / day, about 5 to about 350 μg / kg / day, about 10 to about 500 μg / kg / day, about 15 to about 850 μg / kg / day, or about 15 to about 350 μg / kg / day. In other embodiments, the effective amount is from about 1 to about 30 μg / kg / day, from about 5 to about 30 μg / kg / day, from about 5 to about 25 μg / kg / day, from about 5 to about 20 μg / kg / day. , About 10 to about 20 μg / kg / day, or about 15 to about 20 μg / kg / day. In suitable embodiments, the dosage is about 15 μg / kg / day, about 20 μg / kg / day, about 25 μg / kg / day, about 30 μg / kg / day, about 35 μg / kg / day, about 40 μg / kg / day. About 50 μg / kg / day, about 100 μg / kg / day, about 150 μg / kg / day, about 250 μg / kg / day, or about 500 μg / kg / day.

  In some embodiments, an effective amount of the arsenic composition sufficient to achieve a protective effect in humans is about 5 to about 200 μg / kg / day, about 10 to about 150 μg / kg / day, about 15 To about 150 μg / kg / day, about 30 to about 150 μg / kg / day, about 30 to about 125 μg / kg / day, about 30 to about 100 μg / kg / day, about 30 to about 85 μg / kg / day, about 15 To about 50 μg / kg / day; about 1 to about 125 μg / kg / day, about 1.5 to about 125 μg / kg / day, about 3 to about 125 μg / kg / day, about 1.5 to about 62.5 μg / day kg / day, about 1 to about 40 μg / kg / day, or about 2 to about 85 μg / kg / day. In some embodiments, a suitable total amount of one or more arsenic compounds is about 31 to about 125 μg / kg / day, about 31 to about 85 μg / kg / day, about 35 to about 80 μg / kg / day, Or in the range of about 40 to about 70 μg / kg / day.

Examples of arsenic compounds that may be used include arsenic oxide (III) (arsenic trioxide (As ₂ O ₃ )), arsenic oxide (V) (As ₂ O ₅ ), arsenic selenide (III) (As ₂ Se ₃ ), Arsenic sulfide (II) (As ₂ S ₂ ), arsenic sulfide (III) (As ₂ S ₃ ), arsenic sulfide (V) (As ₂ S ₅ ), arsenic telluride (III) (As ₂ Te ₃ ) , Sodium arsenate (Na ₂ HAsO ₄ ), sodium arsenite (NaAsO ₂ ), potassium arsenate (KH ₂ AsO ₄ ), sodium arsenyl tartrate (NaC ₄ H ₄ AsO ₆ ), tetraarsenic tetrasulfide (As ₄ S ₄ ), and other arsenic derivatives, including but not limited to.

  In some embodiments, arsenic trioxide inhibits, reduces, or prevents damage to non-cancerous cells in a human subject during radiation or chemotherapeutic treatment of cancer cells in a human subject. Can be used for Arsenic trioxide can be administered in an amount ranging from about 1 μg / kg / day to about 125 μg / kg / day. In some embodiments, arsenic trioxide can be administered in an amount ranging from about 31 μg / kg / day to about 125 μg / kg / day. In some embodiments, arsenic trioxide can be administered in an amount ranging from about 31 μg / kg / day to about 85 μg / kg / day. In various embodiments, the arsenic trioxide is at least 5%, at least 10%, at least 15%, at least 20%, at least 25% compared to a control subject or control subject population that has not been administered arsenic trioxide, At least 30%, at least 40%, at least 50%, or more can inhibit, reduce, or prevent damage to non-cancerous cells.

  In some embodiments, sodium arsenite inhibits, reduces, or prevents damage to non-cancerous cells in a human subject during radiation or chemotherapeutic treatment of cancer cells in a human subject. Can be used to Sodium arsenite can be administered in an amount ranging from about 1 μg / kg / day to about 125 μg / kg / day. In some embodiments, sodium arsenite can be administered in an amount ranging from about 31 μg / kg / day to about 125 μg / kg / day. In some embodiments, sodium arsenite can be administered in an amount ranging from about 31 μg / kg / day to about 85 μg / kg / day. In some embodiments, sodium arsenite can be administered in an amount ranging from about 31 μg / kg / day to about 65 μg / kg / day. In various embodiments, the sodium arsenite is at least 5%, at least 10%, at least 15%, at least 20%, at least 25, compared to a control subject or control population that did not receive sodium arsenite. %, At least 30%, at least 40%, at least 50%, or more can inhibit, reduce, or prevent damage to non-cancerous cells.

  In some embodiments, arsenic trioxide is used to inhibit, reduce, or prevent side effects caused by chemotherapeutic agents and / or radiation therapy. Following administration of arsenic trioxide, radiation therapy or one or more chemotherapeutic agents can be administered to the human subject. In some embodiments, arsenic trioxide is given to the human subject at least one day prior to administration of radiation or one or more chemotherapeutic agents to the human subject. In some embodiments, arsenic trioxide is given to the human subject for at least 3 days prior to administration of the radiation or chemotherapeutic agent to the human subject. In various embodiments, the arsenic trioxide is at least 5%, at least 10%, at least 15%, at least 20%, at least 25% compared to a control subject or control subject population that has not been administered arsenic trioxide, At least 30%, at least 40%, at least 50%, or more can inhibit, reduce or prevent one or more side effects.

  In one aspect, a method for treating a disease state in a subject by administering a treatment having concomitant toxicity with administration of one or more protective doses of arsenic prior to the time of administration of the treatment is disclosed. Is done. Administration of a protective amount of an arsenic-containing compound is accompanied by amelioration of one or more side effects associated with chemotherapy or radiation therapy. For example, chemotherapy and radiation therapy used to treat cancer and certain immunological disorders can cause pancytopenia or a combination of anemia, neutropenia, and thrombocytopenia. Thus, the increase or replacement of hematopoietic cells is often critical to the success of such treatment.

  The effect of administering arsenic to a subject is a reduction in treatment toxicity (eg, hematopoietic toxicity), thus allowing high dose and dose high density protocols to be utilized in the treatment plan for a particular subject. . The overall benefit of practicing this embodiment is that administration of arsenic to a subject reduces or reduces the toxicity of a treatment, such as hematopoietic toxicity. Consequently, the maximum therapeutically acceptable dose for various treatment modalities can be varied. In some cases, the maximum therapeutically acceptable dose of treatment can be increased relative to the maximum therapeutically acceptable dose for a subject who has not been administered arsenic prior to treatment.

  In one embodiment, administration of a protective amount of arsenic is associated with an improvement in bone marrow status following exposure to chemotherapy or radiation therapy. The bone marrow status for the subject is improved if one or more of the following occurs: the density of bone marrow cells in the subject after cancer cell therapy (eg, radiation therapy and / or chemotherapy) Higher than if no pretreatment with the subject arsenic was performed prior to cancer cell therapy. The density of progenitor cells or stem cells in the bone marrow in a subject after cancer cell therapy is higher than if the subject was not pretreated with arsenic prior to cancer cell therapy. The mass of bone marrow tissue in a subject after cancer cell therapy (eg, radiation therapy and / or chemotherapy) is higher than if the subject was not pretreated with arsenic prior to cancer cell therapy. Alternatively, the rate of bone marrow cell proliferation in a subject after cancer cell therapy (eg, radiation therapy and / or chemotherapy) is greater than if the subject was not pretreated with arsenic prior to cancer cell therapy. Efficacy determination can be made at any time after treatment with a chemotherapeutic or radiological agent. For example, efficacy determinations can be made one day, two days, three days, four days, five days, six days, seven days, eight days, nine days after the agent (eg, chemotherapeutic agent or radiation) is delivered to the subject. This can be done after 10 days, or after. Bone marrow samples can be taken from any part of the bone marrow in the subject's body for this analysis. To compare the results with the expected results if no pretreatment with arsenic was performed, the data is one or more that receive the same or similar chemotherapy or radiation therapy but no arsenic From several control individuals, preferably from several individuals. Suitably, the control population is to be treated for the same condition, eg, for the same type of cancer. The amount is a protective amount if any benefit occurs as compared to the control population. For example, a bone marrow condition parameter (eg, a given value or mean value that is closer to a normal value (eg, higher than a value or mean value from a control individual or control group) than a value or mean value from a control individual or control group) If the treated individual or group has a progenitor cell type growth rate), the bone marrow status of the treated individual or group is improved. Optionally, statistical methods can be applied to this analysis as needed. In various embodiments, the protective effective amount is at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, or more, It can be an amount that improves the bone marrow condition.

  For chemotherapy, and in some cases, radiation therapy, the maximum therapeutically acceptable dose can be determined by monitoring the subject's complete blood count (“CBC”). Damage to the subject's bone marrow can be manifested by a low red blood cell count, low white blood cell count, low platelet count, or a combination thereof. In general, normal red blood cell counts range from about 4.5 million to about 6 million per microliter of blood. A red blood cell count of less than about 4 million per microliter of blood may indicate that the chemotherapeutic dose and / or radiation dose is higher than the maximum therapeutically acceptable dose. In general, normal white blood cell counts range from about 4,000 to about 11,000 per microliter of blood. A leukocyte count lower than about 3,500 per microliter of blood may indicate that the chemotherapeutic dose and / or radiation dose is higher than the maximum therapeutically acceptable dose. In general, a normal platelet count ranges from about 150,000 to about 400,000 cells per microliter of blood. Platelet cell counts lower than about 50,000 per microliter of blood can indicate that the chemotherapeutic dose and / or radiation dose is higher than the maximum therapeutically acceptable dose. Methods for determining the maximum therapeutically acceptable dose are described by Gurey, “How to calculate the dose of chemotherapeutic”, British Journal of Cancer (2002), 86: 1297, incorporated herein by reference. 1302.

  Accordingly, in some embodiments, the maximum amount that is therapeutically acceptable can be determined by monitoring a subject's CBC. The amount of chemotherapeutic agent that can be used before the one or more cell counts are lowered below a generally acceptable level is one or more arsenic compounds prior to administration of the one or more chemotherapeutic agents It has been found that this can be increased by administration to a subject. In general, increasing the amount of chemotherapeutic agent administered to a subject increases the effectiveness of the treatment (eg, increases the rate at which cancer cells are destroyed and / or decreases the rate of recurrence) ).

  During radiation therapy, the radiation dose that can be used before one or more cell counts are lowered below a generally acceptable level is the subject of the one or more arsenic compounds prior to administration to the subject of radiation. It has also been found that it can be increased by administration to. In general, increasing the amount of radiation administered to a subject increases the effectiveness of the treatment (eg, increases the rate at which cancer cells are destroyed and / or decreases the rate of recurrence).

  Other factors may be used to determine the maximum therapeutically acceptable dose. For example, the side effect of chemotherapy and radiation therapy can be injury to the intestinal mucosa. Because damage to the intestinal mucosa can affect the body's ability to absorb nutrients, this can also be a dose limiting side effect of radiation therapy and chemotherapy. In some embodiments, the maximum therapeutically acceptable dose of cancer cell therapy can be a dose that produces an acceptable amount of damage to the intestinal mucosa. In some embodiments, the subject's weight may be monitored to determine the status of the intestinal mucosa. For example, significant weight loss (eg, weight loss during treatment greater than 3 pounds from the subject's starting weight) may indicate unacceptable damage to the intestinal mucosa. The amount of chemotherapeutic agent that can be used before unacceptable damage to the intestinal mucosa is caused can be increased by administration to the subject of one or more arsenic compounds prior to administration of the one or more chemotherapeutic agents. It has been found. It is also possible that the radiation dose that can be used during radiation therapy before unacceptable damage to the intestinal mucosa is caused can be increased by administration of one or more arsenic compounds to the subject prior to administration of radiation to the subject. Has been found.

  Radiotherapeutic agents and factors include radiation and waves that induce DNA damage, such as gamma radiation, X-rays, UV radiation, microwaves, electron emission, radioisotopes, and the like. Treatment can be achieved by irradiating the localized tumor site with the type of radiation described above. All of these factors are most likely to cause extensive damage to DNA, DNA precursors, DNA replication and repair capabilities, and chromosome assembly and maintenance.

  The dose range for X-rays ranges from a daily dose of 50-200 X-rays over a long period (3-4 weeks) to a single dose of 2000-6000 X-rays. The dose range for the radioisotope is highly variable and depends on the half-life of the isotope, the intensity and type of radiation emitted, and the uptake by neoplastic cells.

  Chemotherapeutic agents include agents that directly crosslink to DNA, agents that intercalate into DNA, and agents that cause chromosomal and mitotic abnormalities by affecting nucleic acid synthesis. Examples of chemotherapeutic agents include doxorubicin, daunorubicin, mitomycin, actinomycin D, bleomycin, cisplatin, etoposide, tumor necrosis factor, taxol, vincristine, vinblastine, carmustine, melphalan, cyclophosphamide, chlorambucil, busulfan , Fluorouracil ("5FU"), and lomustine. Any of these agents can be used alone or in combination with other agents after pretreatment of the patient with one or more arsenic compounds. Pretreatment of a patient with one or more arsenic compounds can reduce the intensity and incidence of many of the side effects of the chemotherapeutic agents listed above without significantly inhibiting the efficacy of such agents, Reduce.

  Agents that directly cross-link nucleic acids, particularly DNA, are envisioned to cause DNA damage and have been shown herein, leading to synergistic antineoplastic multi-drug therapy. Agents such as cisplatin and other DNA alkylating agents can be used.

Agents that damage DNA also include compounds that interfere with DNA replication, mitosis, and chromosome segregation. Examples of these compounds include doxorubicin (also known as adriamycin), etoposide (also known as VP-16), verapamil, podophyllotoxin and the like. These compounds are widely used in clinical settings for the treatment of neoplasms, from 25-75 mg / m ² at 21 day intervals intravenously for adriamycin to intravenous or oral for etoposide. Administered by bolus injection at doses ranging from 35-100 mg / m ² .

  Doxorubicin hydrochloride, 5,12-naphthacenedione, (8s-cis) -10-((3-amino-2,3,6-trideoxy-a-L-lyxo-hexopyranosyl) oxy) -7,8,9 , 10-tetrahydro-6,8,11-trihydroxy-8- (hydroxyacetyl) -1-methoxy-hydrochloride (hydroxydaunorubicin hydrochloride, adriamycin) is used in the broad spectrum of antineoplastic. It binds to DNA, inhibits nucleic acid synthesis, inhibits mitosis, and promotes chromosomal abnormalities.

  When administered alone, it is the first-line drug for the treatment of goiter and primary hepatocellular carcinoma. It is an ovarian tumor, endometrial tumor, and breast tumor, bronchogenic oat cell carcinoma, non-small cell lung cancer, gastric adenocarcinoma, retinoblastoma, neuroblastoma, mycosis fungoides, pancreatic cancer, prostate cancer, bladder 31st first for the treatment of cancer, myeloma, diffuse histiocytic lymphoma, Wilms tumor, Hodgkin's disease, adrenal tumor, osteosarcoma soft tissue sarcoma, Ewing sarcoma, rhabdomyosarcoma, and acute lymphocytic leukemia It is a component of selective multidrug therapy. It is an alternative drug for the treatment of islet cell cancer, cervical cancer, testicular cancer, and adrenocortical cancer. It is also an immunosuppressant.

  Doxorubicin is difficult to absorb and is typically administered intravenously. The pharmacokinetics are multicompartmental. The distributed phase has a half-life of 12 minutes and 3.3 hours. The elimination half-life is about 30 hours. 40-50% is secreted into bile. Most of the rest is metabolized in the liver, some become active metabolites (doxorubicinol), but a few percent are excreted in the urine. If liver dysfunction is present, the dose is typically suppressed.

Suitable doses are intravenous, adult, 60-75 mg / m ² at 21-day intervals, or 25-30 mg / m ^{2 on} each of two consecutive days or three consecutive days repeated at 3 or 4 week intervals, Or 20 mg / m ² once a week. The lowest dose should be used in older patients if there is prior myelosuppression caused by previous chemotherapy or neoplastic bone marrow infiltration, or if this drug is combined with other myelopoietic agents It is. The dose should be reduced by 50% if serum bilirubin is between 1.2 mg / dL and 3 mg / dL, 75% if higher than 3 mg / dL. The total lifetime dose should not exceed 550 mg / m ^{2 in} patients with normal cardiac function and should not exceed 400 mg / m ² in those who have received mediastinal irradiation. Alternatively, 30 mg / m ^{2 on} each day for 3 consecutive days repeated every 4 weeks. Exemplary ^{doses, 10mg / m 2, 20mg /} m 2, 30mg / m 2, 50mg / m 2, 100mg / m 2, 150mg / m 2, 175mg / m 2, 200mg / m 2, 225mg / m 2 It may be ^{250mg / m 2, 275mg / m} 2, 300mg / m 2, 350mg / m 2, 400mg / m 2, 425mg / m 2, 450mg / m 2, 475mg / m 2, 500mg / m 2. Of course, all of these doses are exemplary, and any dose between these points is also expected to be useful in the present invention.

  Daunorubicin hydrochloride, 5,12-naphthacenedione, (8S-cis) -8-acetyl-10-((3-amino-2,3,6-trideoxy-aL-lyxo-hexanopyranosyl) Oxy) -7,8,9,10-tetrahydro-6,8,11-trihydroxy-10-methoxy-hydrochloride, also known as servidin, is commercially available. Daunorubicin intercalates into DNA, blocks DNA-directed RNA polymerase, and inhibits DNA synthesis. It can prevent cell division at doses that do not interfere with nucleic acid synthesis.

  In combination with other drugs, it is included in first-line chemotherapy in the acute phase of acute myeloid leukemia (as induction of remission), acute lymphocytic leukemia, and chronic myelocytic leukemia in adults. Oral absorption is poor and it must be given intravenously. The half-life of the distribution is 45 minutes and the elimination half-life is about 19 hours. Its active metabolite, daunorubicinol, has a half-life of about 27 hours. Daunorubicin is mostly metabolized in the liver and secreted into bile (about 40%). The dose must be reduced in liver or kidney failure.

Appropriate dose (base equivalent) is 45 mg / m ² / day for intravenous adults, under 60 years, every 3 or 4 weeks for 1 day, 2 days, or 3 days (30 mg for patients over 60 years) / M ² ), or 0.8 mg / kg / day for 3-6 days every 3 or 4 weeks. No more than 550 mg / m ² should be given in lifetime, except for 450 mg / m ² or less if you have ever received chest irradiation. Children, 25 mg / m ² once a week, except when under 2 years of age or body surface is less than 0.5 m (in which case an adult schedule based on weight is used). It is available in an injection form (base equivalent) 20 mg (as a base equivalent corresponding to 21.4 mg of hydrochloride). Exemplary ^{doses, 10mg / m 2, 20mg /} m 2, 30mg / m 2, 50mg / m 2, 100mg / m 2, 150mg / m 2, 175mg / m 2, 200mg / m 2, 225mg / m 2 It may be ^{250mg / m 2, 275mg / m} 2, 300mg / m 2, 350mg / m 2, 400mg / m 2, 425mg / m 2, 450mg / m 2, 475mg / m 2, 500mg / m 2. Of course, these doses are exemplary, and any dose between these points is also expected to be useful in the present invention.

  Mitomycin (also known as mutomycin and / or mitomycin-C) is an antibiotic isolated from a Streptomyces caespitosus broth that has been shown to have antitumor activity. This compound is heat stable, has a high melting point, and is easily dissolved in an organic solvent.

  Mitomycin selectively inhibits the synthesis of deoxyribonucleic acid (DNA). Guanine and cytosine content correlates with the degree of mitomycin-induced crosslinking. At high concentrations of this drug, cellular RNA and protein synthesis are also suppressed.

  In humans, mitomycin is rapidly cleared from serum after intravenous administration. After 30 mg bolus injection, the time required to reduce the serum concentration by 50% is 17 minutes. After intravenous injection of 30 mg, 20 mg, or 10 mg, the maximum serum concentrations were 2.4 mg / mL, 1.7 mg / mL, and 0.52 mg / mL, respectively. Clearance is mainly brought about by metabolism in the liver, but metabolism also occurs in other tissues. The rate of clearance is inversely proportional to the maximum serum concentration, which may be due to saturation of the degradation pathway.

  About 10% of the mitomycin dose is excreted unchanged in the urine. Since the metabolic pathway saturates at relatively low doses, the percentage of dose excreted in urine increases with increasing dose. In children, excretion of intravenous mitomycin is similar.

Actinomycin D _{(dactinomycin) (50-76-0); C 62 H} 86 N 12 O 16 (1255.43) is an antineoplastic drug that inhibits DNA-dependent RNA polymerase. It is a component of a first-line combination therapy for the treatment of choriocarcinoma, fetal rhabdomyosarcoma, testicular tumors, and Wilms tumor. Tumors that do not respond to systemic treatment may respond to local perfusion. Dactinomycin enhances radiation therapy. It is a secondary (centrifugal) immunosuppressant.

  Actinomycin D is used in combination with primary surgery, radiation therapy, and other drugs, particularly vincristine and cyclophosphamide. Anti-malignant tumor activity has also been confirmed in Ewing tumors, Kaposi sarcomas, and soft tissue sarcomas. Actinomycin D may be effective in women with advanced cases of choriocarcinoma. It also produces a consistent response in combination with chlorambucil and methotrexate in patients with metastatic testicular cancer. Response may also be observed in patients with Hodgkin's disease and non-Hodgkin's lymphoma. Actinomycin D has also been used to suppress immune responses, particularly rejection of kidney transplants.

Half of the dose is excreted intact in the bile, 10% is excreted in the urine, and the half-life is about 36 hours. The drug dose does not cross the blood brain barrier. Actinomycin D is supplied as a lyophilized powder (0/5 mg in each vial). The usual daily dose is 10-15 mg / kg, which is given intravenously for 5 days. Additional courses may be given at 3-4 week intervals if no toxic symptoms are caused. Daily injections of 100-400 mg are given to children for 10-14 days. In other dosing regimens, weekly maintenance doses of 3-6 mg / kg, a total of 125 mg / kg, and 7.5 mg / kg are used. Although it is safer to administer the drug to an intravenous drip tube, direct intravenous injection can be avoided with the precautionary measure of discarding the needle used to withdraw the drug from the vial to avoid a subcutaneous reaction. Is given. Exemplary ^{doses, 100mg / m 2, 150mg /} m 2, 175mg / m 2, 200mg / m 2, 225mg / m 2, 250mg / m 2, 275mg / m 2, 300mg / m 2, 350mg / m 2 It may be ^{400mg / m 2, 425mg / m} 2, 450mg / m 2, 475mg / m 2, 500mg / m 2. Of course, all of these doses are exemplary, and any dose between these points is also expected to be useful in the present invention.

  Bleomycin is a mixture of cytotoxic glycopeptide antibiotics isolated from a strain of Streptomyces verticillus. It is easy to dissolve in water.

  Although the exact mechanism of action of bleomycin is not known, available evidence appears to indicate that the primary mode of action is inhibition of DNA synthesis and some that inhibition of RNA and protein synthesis is lower There is evidence.

  In mice, high concentrations of bleomycin are found in the skin, lungs, kidneys, peritoneum, and lymphatic vessels. In contrast to the low concentrations found in hematopoietic tissue, skin and lung tumor cells have been found to have high concentrations of bleomycin. The low concentration of bleomycin found in the bone marrow may be related to the high levels of bleomycin degrading enzyme found in the tissue.

  In patients with> 35 mL creatinine clearance per minute, the final elimination half-life of bleomycin in plasma or serum is approximately 115 minutes. In patients with <35 mL creatinine clearance per minute, plasma or serum terminal elimination half-life increases exponentially as creatinine clearance decreases. In humans, 60% to 70% of the administered dose is recovered in urine as active bleomycin.

  Bleomycin should be considered palliative therapy. It has been shown to be useful in the management of the following neoplasms, either as a single agent or as a proven combination with other approved chemotherapeutic agents: Squamous cell carcinoma, including the mouth, tongue, tonsils, nasopharynx, oropharynx, sinus, palate, lips, buccal mucosa, gingiva, epiglottis, pharynx), skin, penis, cervix, and vulva. It has also been used to treat lymphomas and testicular cancer.

  Because of the possibility of an anaphylactoid reaction, lymphoma patients should be treated with no more than 2 units for the first 2 doses. If no acute reaction occurs, a standard dosing schedule may be followed.

  Improvement of Hodgkin's disease and testicular tumor is rapid and is observed within 2 weeks. If no improvement is seen by this time, the possibility of improvement is low. Squamous cell carcinoma responds more slowly and can take up to 3 weeks before any improvement is observed.

  Bleomycin may be given by intramuscular, intravenous, or subcutaneous routes.

Cisplatin is widely used to treat cancers such as metastatic testicular or ovarian cancer, advanced bladder cancer, head and neck cancer, cervical cancer, lung cancer, or other tumors. Cisplatin can be used alone or in combination with other agents, 15-20 mg / m < ² >, 5 days every 3 weeks, a total of 3 effective course doses used for clinical application . Exemplary ^{doses, 0.50mg / m 2, 1.0mg /} m 2, 1.50mg / m 2, 1.75mg / m 2, 2.0mg / m 2, 3.0mg / m 2, 4. It can be 0 mg / m ² , 5.0 mg / m ² , 10 mg // m ² Of course, all of these doses are exemplary and any dose between these points is also Is expected to be useful.

  Cisplatin is not absorbed orally and must therefore be delivered intravenously, subcutaneously, intratumorally or intraperitoneally by injection.

  In certain embodiments of the invention, cisplatin is used in combination with emodin or an emodin-like compound in the treatment of non-small cell lung cancer. However, it is clear that combinations of cisplatin and emodin or emodin-like compounds can be used to treat any other neu-mediated cancer.

  Etoposide, also known as VP16, is used primarily in the treatment of testicular tumors in combination with bleomycin and cisplatin and in small cell lung cancer in combination with cisplatin. It is also active against Kaposi's sarcoma associated with non-Hodgkin lymphoma, acute non-lymphocytic leukemia, breast carcinoma, and acquired immune deficiency syndrome (AIDS).

Etoposide is available as a solution (20 mg / ml) for intravenous administration and as a 50 mg liquid capsule for oral use. For small cell lung cancer, the intravenous dose (in combination therapy) can be at most 100 mg / m ² , or at least 2 mg / m ² , routinely 35 mg / m ² , daily for 4 days to 50 mg / m ² . m ² is also used daily for 5 days. If given orally, the dose should be doubled. Thus, the dose for small cell lung cancer may be at most 200-250 mg / m ² . Intravenous doses for testicular cancer (in combination therapy) are 50-100 mg / m ² , daily for 5 days, or 100 mg / m ² , administered 3 times every other day. The cycle of treatment is usually repeated every 3-4 weeks. This drug should be administered slowly during the 30-60 minute infusion to avoid hypotension and bronchospasm, possibly due to the solvent used in the formulation.

  Tumor necrosis factor (TNF; cachectin) is an inflammatory mediator that kills several types of cancer cells, activates cytokine production, activates macrophages and endothelial cells, promotes the production of collagen and collagenase, It is also a mediator of septic shock, a glycoprotein that promotes catabolic reactions, fever, and sleep. Some infectious agents cause tumor shrinkage through stimulation of TNF production. Since TNF can be extremely toxic when used alone in an effective dose, the optimal dosing regime will probably use it at lower doses in combination with other drugs. Its immunosuppressive effect is enhanced by γ-interferon, so the combination may be dangerous. TNF and interferon-α hybrids have also been found to have anti-cancer activity.

Taxol is an experimental mitotic inhibitor isolated from the bark of ash, Taxus brevifolia. It binds to tubulin (at a site different from that used by vinca alkaloids) and promotes microtubule assembly. Taxol has recently been clinically evaluated. It has activity against malignant melanoma and ovarian carcinoma. Maximum dose, 5 days per day 30 mg / ^{m 2,} or a 210~250mg / ^{m 2} given as once every 3 weeks. Of course, all of these doses are exemplary, and any dose between these points is also expected to be useful in the present invention. FIG. 6 is a diagram showing conjugation between a peptide and a liposome carrying the anticancer drug taxol through a polyethylene glycol adapter.

  Vincristine blocks mitosis and results in metaphase arrest. It seems likely that most of the biological activity of this drug can be explained by its ability to specifically bind to tubulin and to block the ability of proteins to polymerize into microtubules. . Through disruption of the mitotic apparatus microtubules, cell division stops in metaphase. The failure to properly separate chromosomes during mitosis probably results in cell death.

  Due to the relatively low toxicity of vincristine to normal bone marrow and epithelial cells, this agent is unusual among anti-neoplastic agents, and it is often included in combination with other myelosuppressive agents.

  Unpredictable absorption after oral administration of vinblastine or vincristine has been reported. At normal clinical dose, the peak concentration of each drug in plasma is about 0.4 mM.

  Vinblastine and vincristine bind to plasma proteins. They are highly concentrated in platelets and to a lesser extent in white blood cells and red blood cells.

  Vincristine has a multiphase pattern of clearance from plasma. The final half-life is about 24 hours. This drug is metabolized in the liver, but no biologically active derivative has been identified. In patients with liver dysfunction, the dose should be reduced. If the concentration of bilirubin in plasma is higher than 3 mg / dl (about 50 mM), a dose reduction of at least 50% is necessary.

Vincristine sulfate is available as a solution for intravenous injection (1 mg / ml). Vincristine, used with corticosteroids, is currently the selected treatment for introducing remission in childhood leukemia. The optimal dose for these drugs appears to be vincristine, intravenous, 2 mg / body surface area m ² , weekly, and prednisolone, oral, 40 mg / m ² daily. Adult patients with Hodgkin's disease or non-Hodgkin lymphoma usually receive vincristine as part of a complex protocol. When used in a MOPP regimen, the recommended dose of vincristine is 1.4 mg / m ² . High doses of vincristine appear to be better tolerated by children with leukemia than by adults who may experience severe neurotoxicity. Administration of this drug at a more frequent or higher dose every 7 days appears to increase toxic symptoms without a proportional improvement in response rate. Precautions to avoid extravasation during intravenous administration of vincristine should also be used. Vincristine (and vinblastine) can be infused into the arterial blood supply of tumors at doses several times greater than those that are comparable in toxicity and can be administered intravenously.

  Vincristine has been effective in Hodgkin's disease and other lymphomas. It appears to be slightly less beneficial than vinblastine when used alone in Hodgkin's disease, but when used with mechloretamine, prednisolone, and procarbazine (the so-called MOPP regimen), it is an advanced stage of the disease ( Preferred treatment for III and IV). In non-Hodgkin lymphoma, vincristine is an important agent, especially when used with cyclophosphamide, bleomycin, doxorubicin, and prednisolone. Vincristine is more useful than vinblastine in lymphocytic leukemia. Beneficial responses have been reported in patients with a variety of other neoplasms, especially Wilms tumor, neuroblastoma, brain tumor, rhabdomyosarcoma, and breast, bladder, male and female reproductive system cancers ing.

The dose of vincristine used is determined by the clinician according to the needs of the individual patient. 0.01-0.03 mg / kg or 0.4-1.4 mg / m ² can be administered, or 1.5-2 mg / m ² can be administered. Alternatively, 0.02 mg / m ² , 0.05 mg / m ² , 0.06 mg / m ² , 0.07 mg / m ² , 0.08 mg / m ² , 0.1 mg / m ² , 0.12 mg / m ² 0.14 mg / m ² , 0.15 mg / m ² , 0.2 mg / m ² , 0.25 mg / m ² can be given as a constant intravenous infusion. Of course, all of these doses are exemplary, and any dose between these points is also expected to be useful in the present invention.

  When cells are incubated with vinblastine, microtubule lysis occurs. Unpredictable absorption after oral administration of vinblastine or vincristine has been reported. At normal clinical dose, the peak concentration of each drug in plasma is about 0.4 mM. Vinblastine and vincristine bind to plasma proteins. They are highly concentrated in platelets and to a lesser extent in white blood cells and red blood cells.

  After intravenous injection, vinblastine has a multiphasic pattern of clearance from plasma. After distribution, the drug disappears from plasma with a half-life of about 1 hour and 20 hours.

  Vinblastine is metabolized in the liver to the biologically active derivative deacetylvinblastine. About 15% of the administered dose is detected intact in urine and about 10% is recovered in the stool after excretion in bile. In patients with liver dysfunction, the dose should be reduced. If the concentration of bilirubin in plasma is higher than 3 mg / dl (about 50 mM), a dose reduction of at least 50% is necessary.

Vinblastine sulfate is available in injectable preparations. This drug is given intravenously. Special precautions must be taken against subcutaneous migration, which can cause painful inflammation and ulcers. This drug should not be injected into limbs with impaired circulation. After a single dose of 0.3 mg / kg body weight, myelosuppression reaches its maximum in 7-10 days. If moderate leukopenia (about 3000 cells / mm ³ ) has not been reached, the weekly dose may be gradually increased by an increment of 0.05 mg / kg body weight. In a regimen designed to cure testicular cancer, vinblastine is used at a dose of 0.3 mg / kg every 3 weeks, regardless of blood cell count or toxicity.

  The most important clinical use of vinblastine is in the curative treatment of metastatic testicular tumors in combination with bleomycin and cisplatin. Beneficial responses where significant improvement can be observed in 50-90% of cases have been reported in various lymphomas, especially Hodgkin's disease. The effectiveness of vinblastine in a high proportion of lymphoma is not diminished even if the disease is refractory to alkylating agents. It is also active in Kaposi's sarcoma, neuroblastoma, and Reteller-Siebe disease (histiocytosis X), as well as breast carcinoma and choriocarcinoma in women.

The dose of vinblastine used is determined by the clinician according to the needs of the individual patient. 0.1-0.3 mg / kg can be administered, or 1.5-2 mg / m ² can also be administered. ^{Alternatively, 0.1mg / m 2, 0.12mg /} m 2, 0.14mg / m 2, 0.15mg / m 2, 0.2mg / m 2, 0.25mg / m 2, 0.5mg / m 2 ^{, 1.0mg / m 2, 1.2mg /} m 2, 1.4mg / m 2, 1.5mg / m 2, 2.0mg / m 2, 2.5mg / m 2, 5.0mg / m 2, 6 mg / m ² , 8 mg / m ² , 9 mg / m ² , 10 mg / m ² , 20 mg / m ² can be provided. Of course, all of these doses are exemplary, and any dose between these points is also expected to be useful in the present invention.

  Carmustine (sterile carmustine) is one of the nitrosoureas used to treat certain neoplastic diseases. It is 1,3 bis (2-chloroethyl) -1-nitrosourea. It is a lyophilized light yellow flake or agglomerated mass with a molecular weight of 214.06. It is highly soluble in alcohol and lipids and is difficult to dissolve in water. Carmustine is administered by intravenous infusion after reconstitution, as recommended. Sterile carmustine is generally available in 100 mg single dose vials of lyophilized material.

  Although it is generally agreed that carmustine alkylates DNA and RNA, it is not cross-resistant with other alkylating agents. Like other nitrosoureas, it can also inhibit several important enzymatic processes by carbamoylation of amino acids in proteins.

  Carmustine is a single agent or other approved chemotherapy in brain tumors such as glioblastoma, brainstem glioma, medullobladioma, astrocytoma, ependymoma, and metastatic brain tumor In established combination therapy with drugs, indicated as palliative therapy. It has also been used in combination with prednisolone to treat multiple myeloma. Carmustine is useful as a second-line therapy in combination with other approved drugs in the treatment of Hodgkin's disease and non-Hodgkin's lymphoma, in patients who relapse while being treated with first-line therapy, or in patients who do not respond to first-line therapy. Proven to be.

The recommended dose of carmustine as a single agent in previously untreated patients is 150-200 mg / m ² intravenously every 6 weeks. This may be given as a single dose or divided into daily injections such as 75-100 mg / m ² for 2 consecutive days. If carmustine is used in combination with other myelosuppressive drugs or in patients with depleted bone marrow reserves, the dose should be adjusted accordingly. The dose after the initial dose should be adjusted according to the patient's hematological response to the preceding dose. Other doses such as 10 mg / m ² , 20 mg / m ² , 30 mg / m ² , 40 mg / m ² , 50 mg / m ² , 60 mg / m ² , 70 mg / m ² , 80 mg / m ² , 90 mg / m ² Of course, it is understood that 100 mg / m ² may be used in the present invention. Those skilled in the art refer to “Remington's Pharmaceutical Sciences”, 17th edition. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. In any case, the person responsible for administration will determine the appropriate dose for the individual subject.

  Melphalan, also known as alkeran, L-phenylalanine mustard, phenylalanine mustard, L-PAM, or L-sarcorycin, is a phenylalanine derivative of nitrogen mustard. Melphalan is a bifunctional alkylating agent that is active against selected human neoplastic diseases. It is chemically known as 4- (bis (2-chloroethyl) amino) -L-phenylalanine.

  Melphalan is the active L-isomer of this compound and was first synthesized in 1953 by Bergel and Stock. The D-isomer, known as medphalan, is less active against certain animal tumors and the dose required to produce an effect on the chromosome is greater than that required for the L-isomer. The racemic (DL-) form is known as merphalan or sarcolicin. Melphalan is insoluble in water and has a pKa of about 2.1. Melphalan is available in the form of tablets for oral administration and has been used to treat multiple myeloma.

  Available evidence suggests that about one-third of patients with multiple myeloma show a favorable response to oral administration of this drug.

  Melphalan has been used to treat ovarian epithelial cancer. One commonly utilized regimen for the treatment of ovarian cancer is to administer melphalan as a single course at a dose of 0.2 mg / kg for 5 days daily. The course is repeated every 4 or 5 weeks depending on hematologic tolerance. Alternatively, the dose of melphalan used is at least 0.05 mg / kg / day, or at most 3 mg / kg / day, or any dose between these doses, or higher than these doses possible. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. In any case, the person responsible for administration will determine the appropriate dose for the individual subject.

Cyclophosphamide is 2H-1,3,2-oxazaphosphorin-2-amine, N, N-bis (2-chloroethyl) tetrahydro-, 2-oxide monohydrate. It is available from Mead Johnson under the name Cytoxan and from Adria under the name Neosar. Cyclophosphamide is prepared by diethylane catalysis under the action of triethylamine in 3-amino-1-propanol and N, N-bis (2-chloroethyl) phosphoramidic acid dichloride ((ClCH ₂ CH ₂ ) .sub.2N— Prepared by condensing POCl ₂ ). The condensation is two involving both hydroxyl and amino groups, thus leading to cyclization.

  Unlike other β-chloroethylaminoalkylating agents, it does not readily cyclize to the active ethyleneimonium form until activated by liver enzymes. Thus, the substance is stable and well tolerated in the gastrointestinal tract, is effective by oral and parenteral routes, and does not cause local blistering, necrosis, phlebitis, or even pain.

Suitable doses for adults include 1 to 5 mg / kg / day (usually in combination) or 1-2 mg / kg / day orally depending on gastrointestinal tolerance; 40-50 mg / kg in divided doses over 5 days, or 10-15 mg / kg, or 3-5 mg / kg every 7-10 days, twice a week, or 1.5-3 mg / kg / day . A dose of 250 mg / kg / day may be administered as an antineoplastic agent. Because of the deleterious effects of the gastrointestinal tract, the intravenous route is preferred for loading. During maintenance, a white blood cell count of 3000-4000 / mm ³ is usually desirable. This drug may also be administered intramuscularly, by infiltration or into body cavities. It is available in 100 mg, 200 mg, and 500 mg injectable dosage forms, and in 25 mg and 50 mg tablets. For details regarding doses for administration, those skilled in the art should refer to “Remington's Pharmaceutical Sciences” 15th edition, Chapter 61, which is incorporated herein by reference.

  Chlorambucil (also known as Leukeran) is a nitrogen mustard type bifunctional alkylating agent that has been found to be active against selected human neoplastic diseases. Chlorambucil is chemically known as 4- (bis (2-chloroethyl) amino) benzenebutanoic acid.

Chlorambucil is available in the form of tablets for oral administration. It is absorbed quickly and completely from the gastrointestinal tract. After a single oral dose of 0.6-1.2 mg / kg, the peak level of plasma chlorambucil is reached within 1 hour and the final half-life of the parent drug is estimated at 1.5 hours. 0.1-0.2 mg / kg / day or 3-6 mg / m ² / day, or alternatively 0.4 mg / kg can be used for antineoplastic treatment. Treatment plans are well known to those skilled in the art and can be found in “Physicians Desk Reference” and “Remington's Pharmaceutical Sciences” referenced herein.

  Chlorambucil is indicated for the treatment of malignant lymphoma including chronic lymphocytic (lymphocytic) leukemia, lymphosarcoma, giant follicular lymphoma, and Hodgkin's disease. It is not curative in any of these disorders and can result in clinically useful relief.

  Busulfan (also known as milleran) is a bifunctional alkylating agent. Busulfan is chemically known as 1,4-butanediol dimethanesulfonate.

  Busulfan is not a structural analog of nitrogen mustard. Busulfan is available in the form of tablets for oral administration. Each split tablet contains 2 mg busulfan and the inactive ingredients magnesium stearate and sodium chloride.

  Busulfan is indicated for palliative treatment of chronic myelogenous (myeloid, myelocytic, granulocytic) leukemia. Although not curative, busulfan reduces the total mass of granulocytes, reduces the symptoms of the disease, and improves the patient's clinical status. Approximately 90% of adults with chronic myelogenous leukemia that have not been previously treated will obtain hematological remission with the regression or stabilization of organ hypertrophy after use of busulfan. It has been shown to be superior to spleen irradiation in maintaining survival and hemoglobin levels, and equivalent to irradiation in controlling splenomegaly.

  Fluorouracil (“5FU”) (sold under the trade names Adrucil, Carac, Efudex, and Fluoroplex) is a drug that is a pyrimidine analog used in the treatment of cancer. Some of its main uses are in colorectal cancer and pancreatic cancer. It may also be used to treat inflammatory breast cancer.

Lomustine is one of the nitrosoureas used for the treatment of certain neoplastic diseases. It is 1- (2-chloroethyl) -3-cyclohexyl-1-nitrosourea. It is a yellow powder with an empirical formula of C ₉ H ₁₆ ClN ₃ O ₂ and a molecular weight of 233.71. Lomustine is soluble in 10% ethanol (0.05 mg / mL) and in absolute alcohol (70 mg / mL). Lomustine is relatively insoluble in water (<0.05 mg per mL). It is relatively unionized at physiological pH. Inactive ingredients in lomustine capsules are magnesium stearate and mannitol.

  Although it is generally agreed that lomustine alkylates DNA and RNA, it is not cross-resistant with other alkylating agents. Like other nitrosoureas, it can also inhibit several important enzymatic processes by carbamoylation of amino acids in proteins.

Lomustine may be given orally. After oral administration of radioactive lomustine at doses ranging from 30 mg / m ² to 100 mg / m ² , about half of the given radioactivity was excreted in the form of degradation products within 24 hours.

  The serum half-life of the metabolite ranges from 16 hours to 2 days. Tissue levels are comparable to plasma levels 15 minutes after intravenous administration.

  Lomustine is a single agent in addition to other treatment modalities or other approved chemistries in patients already undergoing appropriate surgical and / or radiation therapy methods for both primary and metastatic brain tumors. It has been shown to be useful in established combination therapies with therapeutic agents. It has also proven effective in second-line therapy for Hodgkin's disease in combination with other approved drugs in patients who relapse while being treated with first-line therapy or in patients who do not respond to first-line therapy .

The recommended dose of romustine in adults and children as a single agent in previously untreated patients is 130 mg / m ² as a single oral dose every 6 weeks. In individuals with impaired bone marrow function, the dose should be reduced to 100 mg / m ² every 6 weeks. If lomustine is used in combination with other myelosuppressive drugs, the dose should be adjusted accordingly. If determined by the clinician to be necessary for the individual to be treated, other doses such as 20 mg / m ² , 30 mg / m ² , 40 mg / m ² , 50 mg / m ² , 60 mg / m It is understood that m ² , 70 mg / m ² , 80 mg / m ² , 90 mg / m ² , 100 mg / m ² , 120 mg / m ² , or any dose between these values can be used.

  Surgical treatment for removal of cancer growth is another standard method for tumor and cancer treatment. This attempts to eliminate the entire cancer growth. However, surgery is generally combined with chemotherapy and / or radiation therapy to ensure destruction of any remaining neoplastic or malignant cells. Thus, surgery can be used in addition to using radiation therapy and chemotherapy to treat tumors.

  Induction of DNA damage is a major mode of action for both radiation therapy and chemotherapy to kill cancer cells, which also strongly activates p53. Based on numerous evidences, the acute toxicity induced by DNA damaging anti-cancer treatment is mainly mediated by p53, which when activated, intestinal epithelium, spleen, bone marrow, thyroid, tongue, testis, And induces massive apoptotic cell death in susceptible tissues including hair follicles, with serious pathological consequences. Consistent with these observations, there is an observation that cells with defective p53 are resistant to DNA damage-induced apoptosis. In addition, genetic studies have shown that p53-deficient mice are refractory to toxicity induced by radiation and chemotherapy. The p53-mediated pathological response to chemotherapy and radiation therapy makes suppression of p53 a possible approach for remission of adverse side effects, and patients are much more aggressive (and hence potentially It suggests that it may be possible to tolerate a more successful treatment plan. However, p53 is one of the most important tumor suppressors and therefore the potential cancer risk resulting from its inhibition needs to be addressed.

  p53 tumor suppressor is a transcription factor that regulates the expression of several genes whose products mediate cell cycle arrest, DNA repair, senescence, or apoptosis. An important role of p53 in preventing carcinogenesis is its universal inactivation in cancer cells, either through mutations that directly affect the p53 locus or through abnormalities in its normal regulation. Supported by. Since the DNA damage response pathway and the carcinogenic stress pathway come together in p53, both pathways are thought to be essential for the tumor suppressor function of p53. However, recent genetic studies provide strong evidence that the carcinogenic stress pathway rather than the DNA damage pathway is essential for p53-mediated tumor suppression. Using a genetically engineered mouse model where p53 status can reversibly switch between functional and inactive states in vivo, the p53-mediated DNA damage response is independent of tumor suppression, It has been shown to be the cause of pathological consequences. Interestingly, the delay in p53 recovery until the acute DNA damage response subsides retains protection against cancer development, and such protection is dependent on p19ARF. Mouse genetic studies in which endogenous p53 has been replaced by a mutant that cannot be phosphorylated by a DNA damage-activated protein kinase (ATM, ATR, or Chk2) have shown that acute DNA damage responses are p53-mediated tumors It is consistent with the opinion that it may not be necessary for containment. Knock-in mice were not capable of DNA damage-induced apoptosis, but were nevertheless completely protected from cancer development. Together, these studies have shown that temporary suppression of p53 activity can significantly reduce DNA damage-induced cytotoxicity without compromising tumor suppressor function, and as a temporary approach to protecting cancer therapy A rationale for exploring potential p53 inhibition is provided.

  Arsenic is a naturally occurring metalloid that induces oxidative stress by activating NADPH oxidase activity through a Ras-GTPase-dependent mechanism, resulting in an intracellular cluster of active oxygen. Exposure of humans, laboratory animals, and cultured cells to arsenic is associated with a variety of diverse effects. Arsenic is a definite human carcinogen, but there is much debate about the shape of the arsenic response curve, especially at low doses. This argument is further complicated by the fact that the mechanism of arsenic carcinogenesis remains unclear because there has been no consistent success in inducing cancer in animal models through arsenic exposure. Epidemiological studies including low-dose data also show that exposure to arsenic in drinking water at a concentration of less than about 60 ppb (0.8 μM) is associated with a lower bladder or lung cancer risk than control values. ing. However, when absorbed at toxic levels, arsenic causes serious health problems including cancer. For example, in many areas of Bangladesh, high concentrations of arsenic in drinking water are a particular health concern because it correlates with increased cancer rates. However, arsenic has also been shown to produce health benefits at lower doses. For example, during the 18th and early 20th centuries, an inorganic arsenic preparation known as Fowler's Solution (1% potassium arsenite) was used to treat a variety of diseases including skin cancer, hypertension, and arthritis. It was. Arsenic was even applied to the skin to improve facial appearance by women. The concentration-related hierarchy of responses to arsenic is well documented. For example, in human adult foreskin keratinocytes, arsenic treatment for 24 hours at a concentration of 5 μM or less is a transcription factor known to promote cell proliferation and cell survival, nuclear factor-κB (NF-κB). And induction of proliferation accompanied by enhanced activator protein 1 (AP-1) activity. A statistically significant decrease in cell viability was observed at concentrations above 10 μM. In support of the different nature of cellular effects induced by low and high concentrations of arsenic, genomic analysis has shown that low doses (5 μM, non-cytotoxic) and high doses (50 μM, cytotoxic) are almost completely Affect the expression of non-overlapping subsets, consistent with a qualitative switch from a pro-survival biological response at low dose to a pro-death response at high dose. Taken together, the information available indicates a biphasic dose response for arsenic. The effect induced by low-dose arsenic is not only different in magnitude and effect from high-dose arsenic, but also in nature, ie cytoprotective versus cytotoxicity.

  The following examples are included to demonstrate preferred embodiments of the invention. The techniques disclosed in the examples below are representative of the techniques discovered by the inventors to function well in the practice of the present invention and thus constitute a preferred mode for their practice. It should be understood by those skilled in the art that it can be considered. However, one of ordinary skill in the art, in view of the present disclosure, may make many changes to the specific embodiments disclosed and achieve similar or similar results without departing from the spirit and scope of the invention. You should understand that you can still get.

[Low dose arsenic suppresses p53 activity by inducing its cytoplasmic distribution]
Short-term treatment of cells with sodium arsenite at low doses (1-10 μM, 12 hours) is associated with upregulation of Hdm2 and cytoplasmic accumulation of p53. Through the MAPK pathway, low levels of arsenite stimulate P2 promoter-mediated expression of Hdm2, which then promotes P53 ubiquitination and subsequent nuclear export. As a result, the p53 response to genotoxic stress is impaired, as evidenced by the failure of p53 activation and apoptosis in response to UV irradiation or 5FU treatment. The ability of arsenite to interfere with p53 activation is further demonstrated by significant blunting of p53-dependent tissue damage induced by 5FU treatment when mice are given arsenite-containing water.

[Pretreatment of tumor-bearing mice with low doses of arsenic protects normal tissues from chemotherapy and radiation-induced killing but does not protect cancer cells]
In the experiment, lung cancer cell line A549 was used to create a mouse xenograft model to evaluate the effects of arsenic. Athymic nude mice (Balb c nu / nu, 4-6 weeks old) were purchased from Harlan laboratories. Mice were housed under aseptic conditions and maintained on a 12 hour light / 12 hour dark cycle, with food and water supplied ad libitum. Human lung cancer cells A549 (cells as a 50% suspension in Matrigel) as 3 million cells per mouse in a final volume of 100 μl were injected subcutaneously into the right flank of Balb c nude mice. When the average tumor volume reached approximately 100 mm ³ , the mice were randomized into the following groups: Control; Arsenite only; 5FU only; Arsenite and 5FU; X-ray irradiation only; Arsenite and X-ray irradiation. For arsenic pretreatment, mice were given water containing sodium arsenite (as 1.0 mg / L) for 3 days. The mice were then given either intravenous 5FU (30 mg / kg body weight) or 2 Gy total body irradiation (“TBI”) daily for a week. Tumor volume was measured periodically. Tumor volume was calculated using the equation: (volume = length × width × depth × 0.5236 mm ³ ). Body weight was also monitored throughout the experiment. Values are mean ± SE from two independent experiments with a total of 10 mice per group.

  In vehicle group or control group, tumor volume continued to increase with time (see FIGS. 1 and 2). Arsenic treatment did not produce any detectable effect on the growth of transplanted tumors, indicating that such short-term arsenic treatment did not promote or inhibit tumor progression. As expected, daily treatment for 1 week with either 5FU by intravenous treatment (30 mg / kg body weight) or irradiation with 2 Gy as total body irradiation (TBI) resulted in significant tumor regression. Significantly, as demonstrated by the observation that 5FU-induced or tumor-induced tumor regression is indistinguishable between the two groups pretreated with or without arsenic, Treatment had little effect on either 5FU-induced tumor suppression or radiation-induced tumor suppression (FIGS. 1 and 2). Thus, our data indicate that low dose arsenic pretreatment does not detectably affect the efficacy of 5FU and radiation, at least in the human lung cancer xenograft mouse model.

  To test whether low-dose arsenic can protect normal tissue from damage caused by 5FU or irradiation in these tumor-bearing mice, we monitored weight changes during the experimental period. did. The body weight of mice treated as described in the experiment was monitored during the experiment.

  As shown in FIGS. 1 and 2, the body weight of the control mice did not level off until week 7, and then increased slightly. Interestingly, arsenic-treated mice showed little weight loss. In marked contrast, mice treated with either 5FU or irradiation lost significant weight. This is probably caused by treatment toxicity because tumor growth in these animals was almost completely suppressed (see FIGS. 1 and 2). Significantly, such treatment-induced weight loss was effectively prevented by arsenic pretreatment, as demonstrated by minimal loss of body weight in mice given arsenic-containing water.

  To assess the effects of arsenic at the tissue level, we examined the two tissues that are most sensitive to chemotherapy and radiation treatment, small intestinal cells and bone marrow cells. Consistent with observations at the whole animal level, 5FU and radiation treatment were accompanied by severe damage to small intestinal cells and bone marrow cells, and such damage was significantly inhibited by low dose arsenic pretreatment. Taken together, the results show that short-term treatment with low dose arsenic is accompanied by significant protection of normal tissue without compromising the ability to kill 5FU and irradiated cancer cells.

[Duration of arsenic pretreatment to provide maximum protection]
Next, the inventors determined the duration of arsenic pretreatment that could provide maximum protection. We pretreated cancer-bearing mice with 1.0 mg / L arsenic in drinking water for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days. The animals were again treated with 5FU (30 mg / kg body weight) for 1 week daily to monitor body weight changes. The results show that 1 day and 2 days arsenic pretreatment resulted in weaker protection than 3 days pretreatment but there was no further increase in benefit from longer than 3 days pretreatment. Taken together, our data indicate that 3 days of arsenic pretreatment appears to be sufficient as optimal protection.

[Arsenic-mediated p53 inhibition is transient and reversible]
Recognizing the potential cancer risk associated with arsenic, we investigated whether low-dose arsenic-mediated p53 inhibition is reversible. After culturing for 3 days in a medium containing 5 μM sodium arsenite, MCF10A cells, a non-transformed human breast epithelial cell line, were cultured in medium without arsenic for 1 day, 3 days, 5 days. Recovery was performed for 7 days, 9 days, or 11 days and the p53 response to irradiation was evaluated. Radiation-induced p53 activation is reflected by induction of p53 protein abundance and p21 expression, and the cells can only be detected after being cultured in arsenic-free medium for 5 days, and almost completely recovered 7 days after arsenic removal did. Consistent with the Western results, immunostaining revealed p53 nuclear redistribution in cells cultured for 5 days in medium without arsenic. Taken together, the results for cultured cells indicate that arsenic-mediated p53 inhibition is transient and can be reversed completely once arsenic treatment is discontinued.

  Next, we investigated arsenic-mediated regulation of p53 activity in vivo using mice. To achieve this goal, we monitored the p53 response to irradiation using peripheral blood lymphocytes. Mice were given water containing arsenic for 3 days. Whole blood was collected from the animals 1 day, 3 days, 5 days, 7 days, 9 days, or 11 days after arsenic removal. Lymphocytes were isolated by using the Ficoll method. Cells were cultured for 24 hours, then irradiated with a dose of 2 Gy and harvested 3 hours after treatment. Western analysis was performed. Consistent with results from cell-based studies, after removal of arsenic, p53 activation by radiation recovered, albeit at a slightly slower reaction rate. Radiation-induced p53 accumulation and p21 expression was suppressed on day 0 and day 1 after irradiation, partially recovered on day 7 and fully returned to untreated levels on day 9. In summary, our data indicate that low-dose arsenic-induced p53 suppression is transient and reversible.

[Human breast MDA MB-321 xenograft female mouse]
Athymic nude mice (Balb c nu / nu, 4-6 weeks old) were purchased from Harlan laboratories. Mice were housed under aseptic conditions and maintained on a 12 hour light / 12 hour dark cycle, with food and water supplied ad libitum. Human breast cancer cells MDA-MB-231 (cells as a 50% suspension in Matrigel) as 3 million cells per mouse in a final volume of 100 ml were injected subcutaneously into the right flank of Balb c nude mice. When the average tumor volume reached approximately 100 mm ³ , the mice were randomized into the following groups: Control; 5FU only; Arsenite and 5FU. For arsenic pretreatment, mice were given water containing sodium arsenite (as 1.0 mg / L) for 3 days or given intraperitoneal (“IP”) treatment (10 μg / day, 3 days). . The mice were then treated with 5FU intravenously (30 mg / kg body weight) for 1 week daily. Tumor volume was measured periodically. Tumor volume was calculated using the equation: (volume = length × width × depth × 0.5236 mm ³ ). Body weight was monitored throughout the experiment. Values are mean ± SE from two independent experiments with a total of 10 mice per group. The results from this experiment are presented in FIG.

  As expected, tumor volume in the vehicle or control group continued to increase with time (FIG. 3). Daily treatment for 1 week with 5FU (30 mg / kg body weight, iv) resulted in significant tumor regression. Arsenic pretreatment is 5FU induced, as evidenced by the observation that 5FU induced tumor regression is indistinguishable between the two groups pretreated with or without sodium arsenite. There was little effect on tumor suppression (FIG. 3). To test whether low dose arsenic can protect normal tissue from damage caused by 5FU in these tumor-bearing mice, we monitored weight changes during the experimental period. Mice treated with 5FU showed a significant decrease in body weight, in contrast to control mice that showed little loss of body weight. As demonstrated by a minimal decrease in body weight of mice pretreated with arsenic trioxide, 5FU-induced weight loss was almost completely prevented by arsenic pretreatment.

[Human colon cancer xenograft (5FU)]
Athymic nude mice (Balb c nu / nu, 4-6 weeks old) were purchased from Harlan laboratories. Mice were housed under aseptic conditions and maintained on a 12 hour light / 12 hour dark cycle, with food and water supplied ad libitum. Human colon cancer cells SW-480 (cells as a 50% suspension in Matrigel) as 3 million cells per mouse in a final volume of 100 ml were injected subcutaneously into the right flank of Balb c nude mice. When the average tumor volume reached approximately 100 mm ³ , the mice were randomized into the following groups: Control; arsenite only, 5FU only; arsenite and 5FU. For arsenic pretreatment, mice were given water containing sodium arsenite (as 1.0 mg / L) for 3 days. The mice were then treated with 5FU intravenously (30 mg / kg body weight) daily for 1 week. Tumor volume was measured periodically. Tumor volume was calculated using the equation: (volume = length × width × depth × 0.5236 mm 3). Body weight was monitored throughout the experiment. Values are mean ± SE from two independent experiments with a total of 10 mice per group. The results from this experiment are presented in FIG.

  As expected, tumor volume in vehicle or control groups continued to increase with time (FIG. 4). Arsenic treatment did not produce any detectable effect on the growth of transplanted tumors, indicating that short-term treatment with such low doses of arsenic does not promote or inhibit tumor progression. Daily treatment for 1 week with 5FU (30 mg / kg body weight, iv) resulted in significant tumor regression. Arsenic pretreatment is 5FU induced, as evidenced by the observation that 5FU induced tumor regression is indistinguishable between the two groups pretreated with or without sodium arsenite. Little effect on tumor suppression. There was little difference between male and female mice in response to 5FU and arsenic treatment. Thus, our data indicate that short-term, low-dose arsenic pretreatment does not detectably affect the efficacy of 5FU, at least in human colon cancer xenograft mouse models.

  To investigate whether low dose arsenic can protect normal tissue from damage caused by 5FU in these tumor-bearing mice, we monitored weight changes during the experimental period. Mice treated with 5FU showed a significant decrease in body weight, in contrast to control and arsenic-treated mice that showed little weight loss. This is probably caused by treatment toxicity because the tumors in these animals almost completely regressed. Significantly, 5FU-induced weight loss was almost completely prevented by arsenic pretreatment, as demonstrated by minimal weight loss in both male and female mice fed arsenic-containing water It was done.

  To confirm the results of body weight measurements, we examined the effects of arsenic at the tissue level by examining the two tissues most sensitive to chemotherapeutic agent-induced damage, small intestinal cells and bone marrow cells. evaluated. Consistent with observations throughout the animals, 5FU treatment was accompanied by severe damage to small intestinal cells, as evidenced by significant changes in both small intestinal crypt size and morphology. Such damage was significantly ameliorated by low-dose arsenic pretreatment. The protective effect of arsenic is also evident in the bone marrow. Bone marrow cell depletion was clearly observed in 5FU-treated mice, but this myeloid reduction was greatly diminished in arsenic pretreated mice. Taken together, the results show that short-term treatment with low dose arsenic is accompanied by significant protection of normal tissue without compromising the ability to kill 5FU cancer cells.

[C57BL / 6 WT mice and R172P knock-in mice]
Our previous studies have shown that the mechanism underlying low-dose arsenic-mediated suppression of apoptosis is inhibition of p53. A mutant p53-expressing mouse model was used to test whether the observed protective effect of arsenic was mediated by p53 inactivation. 4-6 week old black C57BL / 6 WT mice or p53 R172P knock-in mice were obtained from Dr. Lazona. Mice were housed under aseptic conditions and maintained in a 12 hour light / 12 hour dark cycle, food and water were provided ad libitum, and mice were randomized into the following groups: Control; X-ray irradiation only; Arsenite and X-ray irradiation. For arsenic pretreatment, mice were given water containing arsenic trioxide (as 1.0 mg / L) for 3 days. Thereafter, the mice were irradiated with 2 Gy TBI daily for 1 week. Body weight was monitored throughout the experiment. Values are mean ± SE from two independent experiments with a total of 10 mice per group. The results from this experiment are presented in FIG. In contrast to wild-type littermates, arsenic provides little protection against 5FU-induced toxicity in p53 mutant mice, and arsenic exerts a selective protective effect on healthy normal tissues through suppression of p53 I confirmed the model to demonstrate.

[Dose-dependent study of sodium arsenite pretreatment]
Twelve different doses of sodium arsenite were tested on 6 male mice and 6 female mice per group. A sodium arsenite dose above 3500 μg / kg body weight by itself showed very obvious toxicity. As a result, we tested 7 dose groups from 15 μg / kg to 3500 μg / kg. Mice pretreated with arsenic or without arsenic for 3 days were given whole body irradiation at a dose of 2 Gy. Forty-eight hours after irradiation, animals were captured and blood samples and tissues were collected. We used the number of WBC (including lymphocytes, macrophages, and platelets), p53 activity in the GI tract, and GI form as markers for assessing toxicity. Based on these three parameters, when taken together, a protective arsenic dose range is determined, from 15 μg / kg body weight to 1000 μg / kg body weight. The results are presented in FIG.

[Histological study of cells after pretreatment with arsenic]
Mice pretreated with sodium arsenite were subjected to radiation therapy (2 Gy or 6 Gy) or chemotherapy (cisplatin or 5-FU) to assess damage to various cells. Table 1 shows renal cell status after chemotherapy or radiation therapy with or without arsenate pretreatment.

Table 2 shows the state of bone marrow cells after chemotherapy or radiation therapy with or without arsenate pretreatment.

Table 3 shows the status of spleen cells after chemotherapy or radiation therapy with or without arsenate pretreatment.

Table 4 shows the state of small intestinal cells after chemotherapy or radiation therapy, with or without arsenate pretreatment.

Table 5 shows the various tissue conditions after chemotherapy or radiation therapy with or without arsenate pretreatment.

[Dose conversion from animal studies]
Data collected from studies in mice can be used to determine the appropriate dosage range for use in humans. In one embodiment, the dose range for humans based on mouse studies can be determined using the following equation (1).
Human equivalent dose (mg / kg) = mouse dose (mg / kg) * 0.081
When other animals are used, the dosage ranges are as follows: Reagan-Shaw et al., “Dose translation from animal to human studies revised,” The FASEB Journal, Vol. 22, 2007, which is incorporated herein by reference. Adjustments can be made based on similar equations as taught on pages 659-661.

[Arsenic dose-limiting toxicity study]
Arsenic trioxide is available in sterile injectable solutions. The molecular formula in the solid state is As ₂ O ₃ . Its molecular weight is 197.8 grams. Although its mechanism of action is not fully understood, it is thought to inhibit growth and promote apoptosis in many different cancer cell lines. Consistent with currently used 10 mg (0.15 mg / kg FDA approved dose) delivered intravenously over 2 to 3 hours in 500 ml 5% glucose saline for patients with acute promyelocytic leukemia ) Plasma daily arsenic rapidly at an average peak level of 6.85 μmol / L (range) at t1 / 2α 0.89 ± 0.29 hours and t1 / 2β 12.13 ± 3.31 hours. 5.54 to 7.30 μmol / L). It has also been shown that continuous administration of arsenic did not cause changes in the plasma concentration of arsenic. Metabolism of arsenic trioxide involves reduction of pentavalent arsenic from trivalent arsenic by arsenate reductase, and methylation from trivalent arsenic to monomethylarsonic acid and from monomethylarsonic acid to dimethylarsinic acid by methyltransferase. Including. The main place of methylation appears to be the liver. Arsenic is stored in the liver, kidneys, heart, lungs, and nails. The normal excretion pattern is in urine. There are currently numerous NCI-supported clinical trials of arsenic trioxide in hematological and solid tumors. Although arsenic trioxide is a known human carcinogen, its effects on humans at very low levels are controversial and some data support the protective effect of arsenic against certain cancers doing.

For the hematological toxicity assessment of arsenic trioxide, a comparison of alternating chemotherapy cycles with and without arsenic is used, and the measurement of effects on hematological endpoints is studied using a repeated measures design. The dose of arsenic trioxide determined from the dose escalation is used as a constant dose for each patient for clinical toxicity assessment. Endpoints include changes in hematological parameters (white blood cell count, platelet count, and hematocrit) over time, comparing alternating cycles of chemotherapy. Toxicity assessments are performed only for patients whose in vitro p53 activation is blocked at the dose administered of arsenic trioxide. For these patients, arsenic trioxide is not administered for chemotherapy cycles 1, 3, and 5. Arsenic trioxide is administered for cycles 2, 4, and 6. Each patient receives the same dose of arsenic trioxide prior to an even cycle of chemotherapy.

[Overview]
Induction of DNA damage is a major mode of action for both radiation therapy and chemotherapy to kill cancer cells, which also strongly activates p53. Based on numerous evidences, the acute toxicity induced by DNA damaging anti-cancer treatment is mainly mediated by p53, which when activated, intestinal epithelium, spleen, bone marrow, thyroid, tongue, testis, And induces massive apoptotic cell death in susceptible tissues including hair follicles, with serious pathological consequences. Consistent with these observations, there is an observation that cells with defective p53 are resistant to DNA damage-induced apoptosis. In addition, genetic studies have shown that p53-deficient mice are refractory to toxicity induced by radiation and chemotherapy. The p53-mediated pathological response to chemotherapy and radiation therapy makes suppression of p53 a possible approach for remission of adverse side effects, and patients are much more aggressive (and hence potentially It suggests that it may be possible to tolerate a more successful treatment plan. However, p53 is one of the most important tumor suppressors and therefore the potential cancer risk resulting from its inhibition needs to be addressed.

  p53 tumor suppressor is a transcription factor that regulates the expression of several genes whose products mediate cell cycle arrest, DNA repair, senescence, or apoptosis. An important role of p53 in preventing carcinogenesis is its universal inactivation in cancer cells, either through mutations that directly affect the p53 locus or through abnormalities in its normal regulation. Supported by. Since the DNA damage response pathway and the carcinogenic stress pathway come together in p53, both pathways are thought to be essential for the tumor suppressor function of p53. However, recent genetic studies provide strong evidence that the carcinogenic stress pathway rather than the DNA damage pathway is essential for p53-mediated tumor suppression. Using a genetically engineered mouse model where p53 status can reversibly switch between functional and inactive states in vivo, the p53-mediated DNA damage response is independent of tumor suppression, It has been shown to be the cause of pathological consequences. Interestingly, the delay in p53 recovery until the acute DNA damage response subsides retains protection against cancer development, and such protection is dependent on p19ARF. A mouse genetic study in which endogenous p53 was replaced by a mutant that cannot be phosphorylated by DNA damage-activated protein kinase (ATM, ATR, or Chk2) has shown that acute DNA damage response is p53-mediated tumor suppression It is consistent with the opinion that it may not be necessary for us. Knock-in mice were not capable of DNA damage-induced apoptosis, but were nevertheless completely protected from cancer development. Together, these studies have shown that temporary suppression of p53 activity can significantly reduce DNA damage-induced cytotoxicity without compromising tumor suppressor function, and as a temporary approach to protecting cancer therapy A rationale for exploring potential p53 inhibition is provided.

  Arsenic is a naturally occurring metalloid that induces oxidative stress by activating NADPH oxidase activity through a Ras-GTPase-dependent mechanism, resulting in an intracellular cluster of active oxygen. Exposure of humans, laboratory animals, and cultured cells to arsenic is associated with a variety of diverse effects. Arsenic is a definite human carcinogen, but there is much debate about the shape of the arsenic response curve, especially at low doses. This argument is further complicated by the fact that the mechanism of arsenic carcinogenesis remains unclear because there has been no consistent success in inducing cancer in animal models through arsenic exposure. Epidemiological studies including low-dose data also show that exposure to arsenic in drinking water at a concentration of less than about 60 ppb (0.8 μM) is associated with a lower bladder or lung cancer risk than control values. ing. However, when absorbed at toxic levels, arsenic causes serious health problems including cancer. For example, in many areas of Bangladesh, high concentrations of arsenic in drinking water are a particular health concern because it correlates with increased cancer rates. However, arsenic has also been honed to produce health benefits at lower doses. For example, during the 18th and early 20th centuries, an inorganic arsenic preparation known as Fowler's Solution (1% potassium arsenite) was used to treat a variety of diseases including skin cancer, hypertension, and arthritis. It was. Arsenic was even applied to the skin to improve facial appearance by women. The concentration-related hierarchy of responses to arsenic is well documented. For example, in human adult foreskin keratinocytes, arsenic treatment for 24 hours at a concentration of 5 μM or less is a transcription factor known to promote cell proliferation and cell survival, nuclear factor-κB (NF-κB). And induction of proliferation accompanied by enhanced activator protein 1 (AP-1) activity. A statistically significant decrease in cell viability was observed at concentrations above 10 μM. In support of the different nature of cellular effects induced by low and high concentrations of arsenic, genomic analysis has shown that low doses (5 μM, non-cytotoxic) and high doses (50 μM, cytotoxic) are almost completely Affect the expression of non-overlapping subsets, consistent with a qualitative switch from a pro-survival biological response at low dose to a pro-death response at high dose. Taken together, the information available indicates a biphasic dose response for arsenic. The effect induced by low-dose arsenic is not only different in magnitude and effect from high-dose arsenic, but also in nature, ie cytoprotective versus cytotoxicity.

  While not intending to be bound by any particular theory, low-dose arsenic pretreatment can effectively protect normal tissue from DNA damage-induced pathological consequences through a mechanism of transient p53 suppression. it is conceivable that. Importantly, the observation is that protection is selective to normal tissue and cancer cells do not share this protective response because of their defective p53. We have also established a method using peripheral lymphocytes to monitor the effect of arsenic on p53 activity.

[Example-Effect of low dose arsenic on p53 in human subjects with malignant tumors]
The purpose of this study was to evaluate the activity of arsenic trioxide in blocking p53 activation in human patients. p53 activation was measured by an in vitro assay of patient lymphocytes.

[Method]
Patients older than 18 years who were diagnosed with malignant tumors other than leukemia who were to start chemotherapy known to suppress peripheral blood counts were enrolled in the study. The expected interval between each cycle of chemotherapy was a minimum of 2 weeks. Patients were treated with the first cycle of chemotherapy without arsenic pretreatment, followed by the second cycle of chemotherapy with arsenic pretreatment for 3 consecutive days (arsenic trioxide during cycle 2, day -3, Administered on days 2 and -1). Treatment was given intravenously at a dose of 0.005 mg / kg over 1/2 hour.

  The effect of arsenic on p53 inhibition was measured by p53 activation on day 1/1 of the cycle (at which time the patient received no arsenic or chemotherapy) and cycle 2 when the patient received arsenic pretreatment for 3 days. / Determined by comparison with p53 activation on day 1. Patients whose peripheral lymphocytes did not show p53 activation inducible by irradiation with 2 Gy in culture on cycle 1/1 day were not considered in the final results for this example.

  For the p53 activation assay, approximately 10 mL of whole blood was isolated from the patient on the first day of each cycle. Lymphocytes were then separated on a Ficoll gradient, cells were washed with PBS, divided into two aliquots, and incubated in medium at 37 ° C. prior to assay. Next, one aliquot was exposed to 2 Gy radiation to induce p53 activation. The other aliquot was not processed. Cells were harvested, lysed, and p53 activation was measured using PathScan® Phospho-p53 (Serl5) Sandwich ELISA kit (Cell Signaling, Technology, Danvers, Mass.).

  Briefly, this assay is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-p53 (Ser15) protein. p53 rabbit monoclonal antibody was coated on the microwells. After incubation with cell lysate, both non-phospho-p53 protein and phospho-p53 protein were captured by the coated antibody. After extensive washing, phospho-p53 (16G8) mouse monoclonal antibody was added to detect the captured phospho-p53 protein. Thereafter, the bound detection antibody was recognized using an HRP-linked anti-mouse antibody. HRP substrate TMB was added for color development. The magnitude of the optical density for this color development is proportional to the amount of phospho-p53 protein. P53 activation in treated (irradiated) and untreated samples was compared by examining the difference in optical density between the two samples. The difference in optical density between the treated and untreated samples was divided by the optical density of the untreated sample to obtain the percentage difference (ELISA percentage).

[result]
A preliminary cohort of 10 patients was studied. Patients included in this analysis are defined as patients completing at least one cycle of arsenic trioxide paired with a cycle without arsenic trioxide. One of 10 patients in the cohort (subject # 007) showed p53 activation inducible by irradiation with 2 Gy during culture on day 1/1 of the cycle, induced by the patient's lymphocytes It was shown to have p53. After arsenic pretreatment (cycle 2/1 day), the subject showed a decrease in p53 activation inducible by irradiation with 2 Gy during culture. The results for object # 007 are shown in Table 6 below.

  This result shows that at cycle 1/1 day the patient showed strong p53 induction. When patients received arsenic trioxide before the start of the next cycle of chemotherapy, the induction of p53 was greatly suppressed. It is implied that after in vitro exposure to 2 Gy radiation, there will be a close agreement between blocking p53 activation and protecting cell survival in the patient's peripheral lymphocytes. As such, low dose arsenic is useful in methods for protecting normal tissues of human subjects from the effects of chemotherapy in vivo.

  In this patent, certain US patents, US patent applications, and other materials (eg, articles) are incorporated by reference. However, the texts of such U.S. patents, U.S. patent applications, and other materials are referenced to the extent that there is no discrepancy between such text and the other descriptions and drawings presented herein. Is only incorporated by In the event of such a conflict, any such conflicting text in U.S. patents, U.S. patent applications, and other materials incorporated by such reference is not strictly incorporated by reference in this patent. .

  Further modifications and alternative embodiments of various aspects of the invention will be apparent to those skilled in the art in view of this description. Accordingly, this description is to be construed as illustrative only and is for the purpose of teaching those skilled in the art the general manner of carrying out the invention. It is to be understood that the forms of the invention shown and described herein are to be taken as examples of embodiments. Elements and materials may be substituted for those illustrated and described herein, parts and processes may be reversed, and certain features of the invention may be utilized independently, All will be apparent to the skilled person after enjoying the benefits of this description of the invention. Changes may be made in the elements described herein without departing from the spirit and scope of the invention as described in the following claims.

Claims (20)

For human subjects in need of radiation treatment and / or chemotherapeutic treatment for the treatment of cancer, one or more arsenic compounds in protective amounts from 1 μg / kg / day to 125 μg / kg / day Administering one or more of said arsenic compounds to a human subject at least one day prior to administration of radiation and / or one or more chemotherapeutic agents to the human subject; ,
Administering radiation and / or one or more chemotherapeutic agents to the human subject after one or more administrations of the arsenic compound, and radiotherapy and / or chemotherapeutic treatment of cancer cells in the human subject. A method of inhibiting, preventing or reducing damage to non-cancerous cells in a human subject during treatment.   The method of claim 1, wherein the one or more arsenic compounds comprises arsenic trioxide.   The method of claim 1, wherein the protective amount of the one or more arsenic compounds is from 31 μg / kg / day to 85 μg / kg / day.   The method of claim 1, wherein the protective amount of the one or more arsenic compounds is from 40 μg / kg / day to 70 μg / kg / day.   2. The method of claim 1, wherein one or more of the arsenic compounds is administered to the human subject at least 3 days prior to administration of radiation to the human subject.   The method of claim 1, wherein one or more of the arsenic compounds is administered to a human subject for at least 3 consecutive days prior to administration of radiation to the human subject.   The method of claim 1, wherein one or more of the arsenic compounds is administered orally to a human subject.   The method of claim 1, wherein one or more of the arsenic compounds is administered intravenously to a human subject.   The method of claim 1, wherein the subject is in need of chemotherapeutic treatment.   The method of claim 1, wherein the subject is in need of radiation treatment. Administering one or more arsenic compounds to a human subject;
Providing the subject with a therapeutically acceptable maximum dose of one or more chemotherapeutic agents after one or more administrations of the arsenic compound, wherein the arsenic compound is administered after the one or more administrations The one or more chemotherapeutic agents, wherein the maximum therapeutically acceptable dose of the one or more chemotherapeutic agents given is given in the absence of pretreatment with one or more arsenic compounds A method of chemotherapeutic treatment of cancer cells in a human subject comprising the step of higher than the therapeutically effective maximum dose of   The method of claim 11, wherein the one or more arsenic compounds comprises arsenic trioxide.   12. The method of claim 11, wherein the protective amount of one or more arsenic compounds is from 1 [mu] g / kg / day to 125 [mu] g / kg / day.   12. The method of claim 11, wherein the protective amount of one or more arsenic compounds is from 1 [mu] g / kg / day to 85 [mu] g / kg / day.   12. The method of claim 11, wherein the protective amount of one or more arsenic compounds is from 31 [mu] g / kg / day to 85 [mu] g / kg / day.   12. The method of claim 11, wherein one or more of the arsenic compounds is administered to a human subject at least 3 days prior to administration of radiation to the human subject.   12. The method of claim 11, wherein one or more of the arsenic compounds is administered to a human subject for at least 3 consecutive days prior to administration of radiation to the human subject.   12. The method of claim 11, wherein the maximum therapeutically acceptable dose is determined by monitoring the subject's CBC.   12. The method of claim 11, wherein the maximum therapeutically acceptable dose is determined by monitoring the subject's body weight. Administering one or more arsenic compounds to a human subject;
Administering to the subject a therapeutically acceptable maximum dose of radiation after one or more administrations of the arsenic compound, the radiation being provided after the one or more administrations of the arsenic compound; A maximum therapeutically acceptable dose of a cancer cell in a human subject comprising: a step higher than a therapeutically effective maximum dose of radiation given in the absence of pretreatment with one or more arsenic compounds. Method of radiation treatment.