Moses R. Repaglinide in combination therapy with metformin in type 2 diabetes. The structure—activity relationship between peroxisomeproliferator-activated. One example of an enhancing amount of metformin is about mg/day. cyclophosphamide, ifosfamide, cytarabine, 6-mercaptopurine, 6-thioguanine, . used herein, refers to compounds with similar chemical structure and similar function. Pharmacokinetic-pharmacodynamic relationships of the anthracycline. A zinc(II) amidine complex: Tandem synthesis, structure and self assembly. Article .. Metformin is currently the first-line drug in the treatment of type II diabetes. . of melphalan are described and structure-activity relationships are discussed.
The invention is further directed to a tumor inhibiting pharmaceutical composition comprising an enhancing amount of metformin and a reduced amount of one or more chemotherapeutic agents, wherein the tumor inhibiting amount of the chemotherapeutic agent s is an amount that results in decreased side effects. In one embodiment of the methods described herein, the chemotherapeutic agent is not inhibitory to cancer stem cells.
In another embodiment of the methods described herein, the chemotherapeutic agent is inhibitory to cancer stem cells. Combination therapies with cancer stem cell inhibitory and non-inhibitory chemotherapeutics is also envisioned.
In one embodiment, the enhancing amount of metformin is administered with a cocktail of standard chemotherapeutic agents e. The effect of co-administration of metformin, as discussed herein, is also expected to enhance other forms of anti-tumor therapies, e.
This includes other agents e. Such agents can be tested in assays e. The ability of metformin to enhance a given chemotherapetuic agent or treatment thereby allowing a reduced amount to be given to a subject, is within the ability of the skilled practitioner.
A Structural Basis for Biguanide Activity.
For example, enhancement of a chemotherapeutic agent or tumor treatment is evidenced by increased efficacy of the agent or treatment when in combination with metformin administration, as compared to one or more appropriate controls lacking metformin administration. Efficacy of treatment can be judged by an ordinarily skilled practitioner. Efficacy can be assessed in animal models of cancer and tumor, for example treatment of a rodent with a cancer, and any treatment or administration of the compositions or formulations that leads to a decrease of at least one symptom of the tumor, for example a reduction in the size of the tumor or a slowing or cessation of the rate of growth of the tumor indicates effective treatment.
Efficacy for any given formulation can also be judged using an experimental animal model of cancer, e. When using an experimental animal model, efficacy of treatment is evidenced when a reduction in a symptom of the tumor, for example a reduction in the size of the tumor or a slowing or cessation of the rate of growth of the tumor occurs earlier in treated, versus untreated animals. Experiments detailed in the Examples section below indicate that metformin selectively kills cancer stem cells, and that this killing occurs when the cancer stem cells are exposed to relatively low concentrations of the metformin.
Such a subject may be, for example, predisposed for tumor development e. Without limitation, examples of such genetic predispositions include predisposing mutations in the brca1, brcaII, rb,or p53 gene. In one embodiment, the subject has previously received chemo or radiation therapy and is at high risk for developing a secondary cancer.
One such example is a subject who was treated for childhood leukemia or lymphoma. In one embodiment, the metformin is administered by the methods described herein, to contact a precancerous lesion e. In another embodiment, the metformin is administered following removal of such a lesion e. Another aspect of the present invention relates to the treatment of bone marrow or peripheral blood bone marrow stem cells samples with metformin prior to autologous transplants in the treatment of blood cancer, to thereby reduce cancer stem cells.
Such treatment will decrease the likelihood of reseeding stem cells. In one embodiment, the metfomin can be administered to the subject receiving the transplant after the transplant has taken place e. Another aspect of the present invention relates to the administration of low doses of metformin for long-term cancer prevention in a subject. In one embodiment, such administration is in the form of a dietary supplement or a regular food supplemented with the metformin e. Such formulations of food and dietary supplements are also encompassed by the present invention.
Another aspect of the present invention relates to an assay for testing derivatives of metformin for the ability to enhance chemotherapeutic agents, tumor killing agents, and other therapies, in killing cancer cells.
The cell assays in described in the Examples section below can be adapted for such assays by the skilled artisan. Dosage and Administration In therapeutic applications, the standard dosages and administration schedule of the chemotherapeutic agent or therapy used can vary depending on a number of variables, such as combinations of cytotoxic agents or therapies being administered, the tumor type, the age, weight, and clinical condition of the recipient patient, the route of administration, and the experience and judgment of the clinician or practitioner administering the therapy.
In another embodiment, the therapeutic amount of metformin e. Administration is performed such that the administered agents e.
Suitable routes of administration are known in the art. The agents described herein may be administered in any manner found appropriate by a clinician, such as those described in the Physicians' Desk Reference, For example, parenterally, enterally, topically. The combined agents, or each agent individually can be administered by any means known in the art. Such modes include oral, rectal, nasal, topical including buccal and sublingualor parenteral including subcutaneous, intramuscular, intravenous, and intradermal administration.SAR of Parasympatholytic agents/ antimuscarinic agents/ anticholinergic agents
The metformin and enhanced agent can be adminsitered systemically, or can be administered locally to the, or near the tumor site e. In one embodiment, the metformin and enhanced therapeutic agents e.
Administration can be pre-operative or post-operative, or both. In one embodiment, the metformin is adminstered three times a day e. Administration of metformin when applicable with a chemotherapeutic agent in the methods described herein can be for extended period of time e. In one embodiment, the metformin is administered more often than the chemotherapeutic agent. For example, a subject can be administered the chemotherapeutic agent s e.
Generally, the dose and administration scheduled should be sufficient to result in slowing, and preferably regressing, the growth of the tumor s and also preferably causing complete regression of the tumor. In some cases, regression can be monitored by a decrease in blood levels of tumor specific markers.
An effective amount of a pharmaceutical agent is that which provides an objectively identifiable improvement as noted by the clinician or other qualified observer. Regression of a tumor in a patient is typically measured with reference to the diameter of a tumor.
Decrease in the diameter of a tumor indicates regression. Regression is also indicated by failure of tumors to reoccur after treatment has stopped.
USA1 - Use of metformin in cancer treatment and prevention - Google Patents
The metformin and chemotherapeutic agents in combination, or separately, are delivered at periodic intervals that can range from several times a day to once per month. As noted above, the agents are administered until the desired therapeutic outcome has been obtained. Additionally, in order to avoid side-effects, not all components of the combination may require delivery at each administration. Therapeutic Agents Currently available cytotoxic drugs can be broadly divided by their mechanism of action into four groups: The choice of a particular cytotoxic agent to treat an individual with cancer is influenced by many factors, including the type of cancer, the age and general health of the patient, and issues of multidrug resistance.
The composition of the invention can utilize a variety of cytotoxic agents, including but not limited to the following agents including possible sources: Preferred cytotoxic agents include cyclophosphamide, ifosfamide, cytarabine, 6-mercaptopurine, 6-thioguanine, doxorubicin, daunorubicin, mitoxantrone, and vincristine.
The most preferred cytotoxic agent are cyclophosphamide and ifosfamide. Chemotherapeutic agents are known in the art and include at least the taxanes, nitrogen mustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas, triazenes; folic acid analogs, pyrimidine analogs, purine analogs, vinca alkaloids, antibiotics, enzymes, platinum coordination complexes, substituted urea, methyl hydrazine derivatives, adrenocortical suppressants, or antagonists.
More specifically, the chemotherapeutic agents may be one or more agents chosen from the non-limiting group of steroids, progestins, estrogens, antiestrogens, or androgens. Even more specifically, the chemotherapy agents may be azaribine, bleomycin, bryostatin-1, busulfan, carmustine, chlorambucil, carboplatin, cisplatin, CPT, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, dexamethasone, diethylstilbestrol, doxorubicin, ethinyl estradiol, etoposide, fluorouracil, fluoxymesterone, gemcitabine, hydroxyprogesterone caproate, hydroxyurea, L-asparaginase, leucovorin, lomustine, mechlorethamine, medroprogesterone acetate, megestrol acetate, melphalan, mercaptopurine, methotrexate, methotrexate, mithramycin, mitomycin, mitotane, paclitaxel, phenyl butyrate, prednisone, procarbazine, semustine streptozocin, tamoxifen, taxanes, taxol, testosterone propionate, thalidomide, thioguanine, thiotepa, uracil mustard, vinblastine, or vincristine.
The use of any combinations of chemotherapy agents is also contemplated. Other suitable therapeutic agents are selected from the group consisting of radioisotope, boron addend, immunomodulator and chemosensitizing agent See, U.
Other suitable chemotherapeutic agents, such as experimental drugs, are known to those of skill in the art. It is well known in the art that various methods of radionuclide therapy can be used for the treatment of cancer and other pathological conditions, as described, e.
In another embodiment, different isotopes that are effective over different distances as a result of their individual energy emissions are used as first and second therapeutic agents.
Such agents can be used to achieve more effective treatment of tumors, and are useful in patients presenting with multiple tumors of differing sizes, as in normal clinical circumstances. Few of the available isotopes are useful for treating the very smallest tumor deposits and single cells. In these situations, a drug or toxin may be a more useful therapeutic agent. Accordingly, in preferred embodiments of the present invention, isotopes are used in combination with non-isotopic species such as drugs, toxins, and neutron capture agents.
Many drugs and toxins are known which have cytotoxic effects on cells, and can be used in connection with the present invention. They are to be found in compendia of drugs and toxins, such as the Merck Index, Goodman and Gilman, and the like, and in the references cited above.
Drugs that interfere with intracellular protein synthesis can also be used in the methods of the present invention; such drugs are known to those skilled in the art and include puromycin, cycloheximide, and ribonuclease.
Radiation Therapy A variety of radiation therapies are used in tumor therapy. Applicants envision the use of enhancing amounts of metaformin to allow reduced amounts of any one or a combination of such radiation therapies in tumor treatment. For some types of tumors, radiation may be given to areas that do not have evidence of tumors. This is done to prevent tumor cells from growing in the area receiving the radiation. This technique is called prophylactic radiation therapy.
Radiation therapy also can be given to help reduce symptoms such as pain from cancer that has spread to the bones or other parts of the body. This is called palliative radiation therapy.
Radiation may come from a machine outside the body external radiationmay be placed inside the body internal radiationor may use unsealed radioactive materials that go throughout the body systemic radiation therapy. The type of radiation to be given depends on the type of cancer, its location, how far into the body the radiation will need to go, the patient's general health and medical history, whether the patient will have other types of cancer treatment, and other factors.
Most people who receive radiation therapy for cancer have external radiation. Some patients have both external and internal or systemic radiation therapy, either one after the other or at the same time. External radiation therapy usually is given on an outpatient basis; most patients do not need to stay in the hospital. External radiation therapy is used to treat most types of cancer, including cancer of the bladder, brain, breast, cervix, larynx, lung, prostate, and vagina.
In addition, external radiation may be used to relieve pain or ease other problems when cancer spreads to other parts of the body from the primary site. Intraoperative radiation therapy IORT is a form of external radiation that is given during surgery. IORT is used to treat localized cancers that cannot be completely removed or that have a high risk of recurring coming back in nearby tissues.
After all or most of the cancer is removed, one large, high-energy dose of radiation is aimed directly at the tumor site during surgery nearby healthy tissue is protected with special shields. The patient stays in the hospital to recover from the surgery. IORT may be used in the treatment of thyroid and colorectal cancers, gynecological cancers, cancer of the small intestine, and cancer of the pancreas.
It is also being studied in clinical trials research studies to treat some types of brain tumors and pelvic sarcomas in adults.
Prophylactic cranial irradiation PCI is external radiation given to the brain when the primary cancer for example, small cell lung cancer has a high risk of spreading to the brain. Internal radiation therapy also called brachytherapy uses radiation that is placed very close to or inside the tumor. The radiation source is usually sealed in a small holder called an implant. Implants may be in the form of thin wires, plastic tubes called catheters, ribbons, capsules, or seeds.
The implant is put directly into the body. Internal radiation therapy may require a hospital stay. Internal radiation is usually delivered in one of two ways, each of which uses sealed implants. Interstitial radiation therapy is inserted into tissue at or near the tumor site. It is used to treat tumors of the head and neck, prostate, cervix, ovary, breast, and perianal and pelvic regions.
Intracavitary or intraluminal radiation therapy is inserted into the body with an applicator. It is commonly used in the treatment of uterine cancer. Researchers are also studying these types of internal radiation therapy for other cancers, including breast, bronchial, cervical, gallbladder, oral, rectal, tracheal, uterine, and vaginal.
Systemic radiation therapy uses radioactive materials such as iodine and strontium The materials may be taken by mouth or injected into the body. Systemic radiation therapy is sometimes used to treat cancer of the thyroid and adult non-Hodgkin lymphoma.
Tumors Tumors to be treated by the methods and compositions of the present invention may be malignant e. Examples of benign tumors for treatment include thyroid adenomas, adrenocortical adenomas, and pituitary adenomas, benign brain tumors e. These tumours are composed of atypical cells, having a capacity for autonomous growth, an imprecise delimitation, an ability to invade neighbouring tissues and vessels and a tendency to disseminate by the production of metastases.
Without limitation, examples of cancers which can be treated by the methods and compositions described herein include bladder cancer, melanoma, breast cancer, non-Hodgkin lymphoma, colon and rectal cancer, pancreatic cancer, endometrial cancer, prostate cancer, kidney renal cell cancer, skin cancer, nonmelanomaleukemia, thyroid cancer, lung cancer, cervical cancer, ovarian cancer, testicular cancer. Primary and metastatic growth of the following tumors can be inhibited by the above-described methods: Since metformin can cross the blood brain barrier, it's administration, according to the methods described herein, can be useful in treating or preventing central nervous system tumors, or preventing the spread of cancers to the central nervous system.
The pharmaceutical compositions of this invention may be in the dosage form of solid, semi-solid, or liquid such as, e. Preferably the compositions are administered in unit dosage forms suitable for single administration of precise dosage amounts.
The compositions may also include, depending on the formulation desired, pharmaceutically-acceptabl-e, nontoxic carriers or diluents, which are defined as vehicles commonly used to formulate pharmaceutical compositions for animal or human administration.
Compositions may be provided as sustained release or timed release formulations. The carrier or diluent may include any sustained release material known in the art, such as glyceryl monostrearate or glyceryl distearate, alone or mixed with a wax.
The controlled delivery can be exercised by selecting appropriate macromolecules for example polyesters, polyamino acids, polyvinyl pyrrolidone, ethylenevinylacetate, methylcellulose, carboxymethylcellulose, and protamine sulfate and the concentration of macromolecules as well as the methods of incorporation in order to control release.
Microencapsulation may also be used. The timed release formulation can provide a combination of immediate and pulsed release throughout the day. The diluent is selected so as not to affect the biological activity of the combination. Examples of such diluents are distilled water, physiological saline, Ringer's solution, dextrose solution, and Hank's solution.
US20120220664A1 - Use of metformin in cancer treatment and prevention - Google Patents
In addition, the pharmaceutical composition of formulation may also include other carriers, adjuvants, emulsifiers such as poloxamers, or nontoxic, nontherapeutic, nonimmunogenic stabilizers and the like. Effective amounts of such diluent or carrier will be those amounts which are effective to obtain a pharmaceutically acceptable formulation in terms of solubility of components, or biological activity, and the like.
Another apsect of the present invention relates to a formulation for treating cancer with the above drug combination. In one embodiment, the formulation includes a controlled-release device where one or several of the drugs are being released in a delayed fashion. Such formulation can be in the form of a tablet or a pill which releases different doses of drugs in different time intervals after being taken orally. Another aspect of the present invention relates to a kit for the treatment of a subject by the methods disclosed herein e.
The kit comprises one or more vials of the metformin and one or more vials of the chemotherapeutic agent s either together or in separate vialsat the doses provided above. The kit may further contain instructions describing their use in combination. The kit may include a formulation of both the metformin together with one or more of the chemotherapeutic agents.
Method for Screening for an Agent that Modulates a Chemotherapeutic Agent or Agents that are Modulated by Metformin The present invention provides for methods to screen for agents e.
In the methods, a metformin derivative is administered with a known chemotherapeutic agent to the cells, and its ability to kill the cells is determined by measuring an indicating parameter of the cells e. The cell viability is compared to an appropriate control which has not received the metformin derivative, and an enhanced killing e.
In the methods, a test compound is administered with metformin or an identified metformin derivative to the cells, and its ability to kill the cells is determined by measuring an indicating parameter of the cells e. The cell viability is compared to an appropriate control which has not received the test compound, and an enhanced killing e.
The test compounds are conveniently added in solution, or readily soluble form, to the medium of cells in culture. The agents may be added in a flow-through system, as a stream, intermittent or continuous, or alternatively, adding a bolus of the compound, singly or incrementally, to an otherwise static solution. In a flow-through system, two fluids are used, where one is a physiologically neutral solution, and the other is the same solution with the test compound added.
The first fluid is passed over the cells, followed by the second. In a single solution method, a bolus of the test compound is added to the volume of medium surrounding the cells.
The overall concentrations of the components of the culture medium should not change significantly with the addition of the bolus, or between the two solutions in a flow through method.
In some embodiments, agent formulations do not include additional components, such as preservatives, that may have a significant effect on the overall formulation. Thus in one embodiment, formulations consist essentially of a test agent and a physiologically acceptable carrier, e.
However, if a compound is liquid without a solvent, the formulation may consist essentially of the compound itself. A plurality of assays may be run in parallel with different agent concentrations to obtain a differential response to the various concentrations. As known in the art, determining the effective concentration of an agent typically uses a range of concentrations resulting from 1: The concentrations may be further refined with a second series of dilutions, if necessary.
Typically, one of these concentrations serves as a negative control, i. The test compound can be any molecule, compound, or other substance which can be administered to a test animal.
In some cases, the test agent does not substantially interfere with animal viability. Suitable test compounds may be small molecules, biological polymers, such as polypeptides, polysaccharides, polynucleotides, and the like. Test compounds can be identified that are therapeutically effective, such as anti-proliferative agents, or as lead compounds for drug development.
In some embodiments, test compound can be from diversity libraries, such as random or combinatorial peptide or non-peptide libraries. Many libraries are known in the art, such as, for example, chemically synthesized libraries, recombinant phage display libraries, and in vivo translation-based libraries.
Examples of chemically synthesized libraries are described in Fodor et al. Saline control was normal and injected intraperitoneally I. Control diabetic group was injected i. Hyperglycemia was induced by a single i. Briefly, rats were weighed and injected with STZ dissolved in a citrate buffer 0. After 72 h blood samples were withdrawn from the retroorbital venous plexus under light ether anesthesia and the plasma was separated by centrifugation for the determination of glucose level.
The treatment was carried out for 30 days after 72 h from STZ injection. At 7 days post-induction of hyperglycemia, blood glucose was assayed by the glucose oxidase method, using a glucometer.
The animals were carefully monitored every day and weighed every week during the experiment. Collection of blood and organs: Blood samples of the fasted rats were collected from the medial retro-orbital venous plexus immediately with capillary tubes Micro Haematocrit Capillaries, Mucaps [ 13 ]. About 9 mL of blood collected in two tubes from each animal, one with EDTA for obtaining plasma, the second was allowed to clot for 30 min.
Then, the blood in two tubes was centrifuged at 3, rpm for 15 min to separate serum and plasma for different biochemical analyses. Triglycerides, cholesterol and high density lipoprotein-cholesterol HDL-c were determined using the commercial kits. Low density lipoprotein-cholesterol LDL-c levels were calculated by using the following formula of Muruganandan et al. Volatile low density lipoprotein-cholesterol VLDL-c levels were calculated by using the following formula of Prakasam et al.
The risk ratio was calculation by dividing the total cholesterol by HDL-c. Antioxidant abilities Assessment of lipid peroxidation as oxidative indicator: Bioindicator in tissues the thiobarbituric acid reactive substances TBARS levels as an index of malondialdehyde MDA production were measured by the method described by Ohkawa et al.
A Structural Basis for Biguanide Activity.
MDA, an end product of lipid peroxidation reacts with TBA-TCA complex to form a colour complex at high temperature exhibiting an absorption maximum at nm. Determination of enzymatic and non-enzymatic antioxidants: Illumination of riboflavin in the presence of O2 and electron donor like methionine generates superoxide anions and this has been used as the basis of ability of SOD.
The reduction of NBT by superoxide radicals to blue colour formazan was followed at nm [ 17 ]. Reduced glutathione level GSH as nonenzymatic antioxidant was estimated based on the method of Beutler et al. Determination of blood glucose level, Hb, HbA1c: Glucose was estimated by O-toluidine method of Sasaki et al.
Hb was estimated by cyanmethaemoglobin method of Drabkin and Austin [ 20 ]. HbA1c was estimated by the method of Sudhakar and Pattabiraman[ 21 ] with modification by Bannon [ 22 ].
Insulin level and C-peptide: Insulin in pancreatic homogenates was determined by Immulite Insulin Diagnostic Products Corporation, Los Angeles which depends on a two-site chemiluminescent enzymelabelled immunometricability [ 23 ] Serum C-peptide was measured by radioimmunoassay Medgenix Diagnostics as described by Kumar et al.
All chemicals and reagents were of pure analytical grade. At time of death, pancreas tissues were dissected, cleared of lymph nodes and fat, blotted, washed from blood and weighed. The pancreas was immediately homogenized in 5 ml cold 2 M acetic acid for 5 s. The extract was centrifuged at 15 r. The third portion of the pancreas was immediately cut into small cubes and transferred to ice-cold fixation buffer 1. The synthesized Cr III complex is leaf green and soluble in dimethylsulfoxide and dimethylformamidepartially soluble in hot methanol and insoluble in water and some other organic solvents.
The slightly electrolytic value may be due to the contribution of the one chloride anion in the outer sphere of chelating skeleton of the Cr III metformin complex. The infrared absorption bands are one of the important tools of analyses used for determining the mode of chelations. The most significant bands of metformin HCl ligand can be classified into two groups: According to the two fundamental vibrational groups mentioned above, the metformin HCl free ligand can be interpreted as follows: