text:
This invention is a method of treating cancer, both carcinomas and sarcomas, and viral infections, in particular HIV through the administration of a pharmaceutical composition containing a benzimidazole derivative. The composition is also claimed. The benzimidazole derivative is selected from the group consisting of:
wherein X is hydrogen, halogen, alkyl of less than 7 carbon atoms or alkoxy of less than 7 carbon atoms; n is a positive integer of less than 4; Y is hydrogen, chlorine, nitro, methyl, ethyl or oxychloro; R is hydrogen, alkylaminocarbonyl wherein the alkyl group has from 3 to 6 carbon atoms, or an alkyl group of from 1 to 8 carbon atoms and R2 is 4-thiazolyl, NHCOOR1 wherein R1 is aliphatic hydrocarbon of less than 7 carbon atoms, prodrugs, pharmaceutically acceptable salts and mixtures thereof and a pharmaceutically acceptable carrier
1. A method for selectively inhibiting growth of colon cancer cells containing abnormal p53, the method comprising administering to a patient in need thereof a therapeutically effective amount of a composition comprising a benzimidazole compound of the formula:
X is hydrogen, halogen, alkyl of less than 7 carbon atoms, or alkoxy of less than 7 carbon atoms;
n is a positive integer of less than 4;
Y is hydrogen, chloro, nitro, methyl, ethyl, or oxychloro;
R is hydrogen, an alkyl group of from 1 to 8 carbon atoms, or alkylaminocarbonyl wherein the alkyl group has from 3 to 6 carbon atoms; and
R2 is NHCOOR1, wherein R1 is an aliphatic hydrocarbon of less than 7 carbon atoms;
or a pharmaceutically acceptable salt thereof, or mixtures thereof, wherein growth of colon cancer cells containing abnormal p53 is selectively inhibited as compared to growth of cells containing normal p53.
3. The method of
4. The method of
5. A method for selectively inhibiting growth of colon cancer cells containing abnormal p53, the method comprising administering to a patient in need thereof a therapeutically effective amount of a composition comprising 2-(methoxycarbonylamino)benzimidazole wherein growth of colon cancer cells containing abnormal p53 is selectively inhibited as compared to growth of cells containing normal p53.
6. The method of
7. The method of
[0001] This is a continuation of application Ser. No. 09/748,651, filed Dec. 22, 2000 which is a divisional application of U.S. Ser. No. 09/264,942, filed Mar. 9, 1999, now U.S. Pat. No. 6,262,093, which is a continuation-in-part application of U.S. Ser. No. 08/ 927,550, filed Sep. 6, 1997, now U.S. Pat. No. 5,880,144, which is a divisional of application U.S. Ser. No. 08/771,193, filed Dec. 20, 1996, now U.S. Pat. No. 5,767,138, and which is a divisional of U.S. Ser. No. 08/420,914, filed Apr. 12, 1995, now abandoned. Application U.S. Ser. No. 09/264,942 is also a continuation-in-part of application U.S. Ser. No. 09/081,384, filed May 19, 1998, now abandoned, and also a continuation-in-part of application U.S. Ser. No. 09/081,627, filed May 19, 1998, now abandoned.
[0002] This invention is a method of treating cancer, both carcinomas and sarcomas, and viral infections, in particular HIV through the administration of a pharmaceutical composition containing a benzimidazole derivative. The composition is also claimed.
[0003] Cancers are the leading cause of death in animals and humans. The exact cause of cancer is not known, but links between certain activities such as smoking or exposure to carcinogens and the incidence of certain types of cancers and tumors has been shown by a number of researchers.
[0004] Many types of chemotherapeutic agents have been shown to be effective against cancers and tumor cells, but not all types of cancers and tumors respond to these agents. Unfortunately, many of these agents also destroy normal cells. The exact mechanism for the action of these chemotherapeutic agents are not always known.
[0005] Despite advances in the field of cancer treatment the leading therapies to date are surgery, radiation and chemotherapy. Chemotherapeutic approaches are said to fight cancers that are metastasized or ones that are particularly aggressive. Such cytocidal or cytostatic agents work best on cancers with large growth factors, i.e., ones whose cells are rapidly dividing. To date, hormones, in particular estrogen, progesterone and testosterone, and some antibiotics produced by a variety of microbes, alkylating agents, and anti-metabolites form the bulk of therapies available to oncologists. Ideally cytotoxic agents that have specificity for cancer and tumor cells while not affecting normal cells would be extremely desirable. Unfortunately, none have been found and instead agents which target especially rapidly dividing cells (both tumor and normal) have been used.
[0006] The development of materials that would target tumor cells due to some unique specificity for them would be a breakthrough. Alternatively, materials that were cytotoxic to tumor cells while exerting mild effects on normal cells would be desirable.
[0007] HIV and other viral infections are another leading cause of death. HIV is a disease in which a virus is replicated in the body which attacks the body's immune system. The HIV virus is not easily destroyed nor is there a good mechanism for keeping the host cells from replicating the virus. Herpes Simplex is another viral infection which is difficult, if not impossible, to cure. A method of treating these diseases and other viral infections is highly desirable. A material which would target the HIV virus and inhibit viral replication is highly desirable.
[0008] The benzimidazole derivatives used herein to treat cancer and/or viral infection have been used as fungicides and as antihelmetics.
[0009] A method of treating cancer, in particular, treating cancers in warm blooded animals and humans, comprising administering a therapeutically effective amount of a composition comprising a benzimidazole compound selected from the group consisting of:
[0010] wherein X is hydrogen, halogen, alkyl of less than 7 carbon atoms or alkoxy of less than 7 carbon atoms; n is a positive integer of less than 4; Y is hydrogen, chlorine, nitro, methyl, ethyl or oxychloro; R is hydrogen, alkylaminocarbonyl wherein the alkyl group has from 3 to 6 carbon atoms, or an alkyl group of from 1 to 8 carbon atoms and R2 is 4-thiazolyl, NHCOOR1 wherein R1 is aliphatic hydrocarbon of less than 7 carbon atoms, and preferably an alkyl group of less than 7 carbon atoms is claimed. Preferably the compositions contain:
[0011] wherein R is an alkyl of 1 through 8 carbon atoms and R2 is selected from the group consisting of 4-thiazolyl, NHCOOR1, wherein R1 is methyl, ethyl or isopropyl and the non-toxic, pharmaceutically acceptable acid salts with both organic and inorganic acids. The most preferred compounds are 2-(4-thiazolyl)benzimidazole, methyl-(butylcarbamoyl)-2-benzimidazolecarbamate and 2-methoxycarbonylamino-benzimidazole and those wherein Y is chloro.
[0012] In the present invention it has been discovered that the compounds described above are useful for the inhibition of HIV and the treatment of HIV infection and similar retrovirus infections. The present invention also provides methods for the treatment of HIV infection comprising administering to a host infected with HIV a pharmaceutically or therapeutically effective or acceptable amount of a compound as described above, particularly those wherein R is 4-thiazolyl.
[0013] The present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound as described above.
[0014] These compositions have been discovered to inhibit the growth of cancer or other tumors in humans or animals and to induce apoptosis of cancer cells by administration of a therapeutically effective amount of the composition, preferably by administering a benzimidazole compound to the site of the cancer.
[0015] More specifically, this invention provides an anti-cancer composition comprising a pharmaceutical carrier and a benzimidazole derivative as defined herein along with a method for treating such cancers. These compositions can induce apoptosis in cancer cells.
[0016] These compositions are also effective against viruses and are used to treat viral infections and this invention provides a method of treating viral infections such as herpes, hepatitis, influenza and rhinoviruses.
[0017] A. Definitions:
[0018] As used herein, a "pharmaceutically acceptable" component is one that is suitable for use with humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a reasonable benefit/risk ratio.
[0019] As used herein, the term "safe and effective amount" refers to the quantity of a component which is sufficient to yield a desired therapeutic response without undue adverse side effects (such as toxicity, irritation, or allergic response) commensurate with a reasonable benefit/risk ratio when used in the manner of this invention. By "therapeutically effective amount" is meant an amount of a compound of the present invention effective to yield the desired therapeutic response. For example to inhibit HIV infection or treat the symptoms of infection in a host or an amount effective to delay the growth of or to cause a cancer, either a sarcoma or lymphoma, to shrink. The specific safe and effective amount or therapeutically effective amount will, vary with such factors as the particular condition being treated, the physical condition of the patient, the type of mammal being treated, the duration of the treatment, the nature of concurrent therapy (if any), and the specific formulations employed and the structure of the compounds or its derivatives.
[0020] As used herein, a "pharmaceutical salts" is salt of the benzimidazole derivatives which are modified by making acid or base salts of the compounds. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids. Preferably the salts are made using an organic or inorganic acid. These preferred acid salts are chlorides, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates, malates, citrates, benzoates, salicylates, ascorbates, and the like.
[0021] As used herein, a "pharmaceutical carrier" is a pharmaceutically acceptable solvent, suspending agent or vehicle for delivering the benzimidazole derivatives to the animal or human. The carrier may be liquid or solid and is selected with the planned manner of administration in mind.
[0022] As used herein, "cancer" refers to all types of cancer or neoplasm or malignant tumors found in mammals, including carcinomas and sarcomas. Examples of cancer are cancer of the brain, breast, cervix, colon, head & neck, kidney, lung, non-small cell lung, melanoma, mesothelioma, ovary, sarcoma, stomach, uterus and Medulloblastoma.
[0023] As used herein, the "benzimidazole derivatives" are the benzimidazoles, and their salts and also their prodrugs. The exact benzimidazoles are described in detail below. The preferred materials are the products sold under the names "Thiabendazole�", "Benomyl�" and "Carbendazim�" by BASF and Hoechst, DuPont and MSD-AgVet.
[0024] As used herein "viruses" includes viruses which infect animals or mammals, including humans. Viruses includes retorviruses, HIV, influenza, polio viruses, herpes, herpes simplex, rhinoviruses, hepatitis, and the like.
[0025] B. The Benzimidazole Derivatives
[0026] The benzimidazole derivatives which are known for their antifungal activities. They are systemic fungicides used to prevent and eradicate fungi. The compounds have the following structure:
[0027] wherein X is hydrogen, halogen, alkyl of less than 7 carbon atoms or alkoxy of less than 7 carbon atoms; n is a positive integer of less than 4; Y is hydrogen, chlorine, nitro, methyl, ethyl or oxychloro; R is hydrogen, alkylaminocarbonyl wherein the alkyl group has from 3 to 6 carbon atoms or an alkyl group having from 1 to 8 carbons, and R2 is 4-thiazolyl, NHCOOR1 wherein R1 is aliphatic hydrocarbon of less than 7 carbon atoms, and preferably and alkyl group of less than 7 carbon atoms. Preferably the compositions are:
[0028] wherein R is an alkyl of 1 through 8 carbon atoms and R2 is selected from the group consisting of 4-thiazolyl, NHCOOR1, wherein R1 is methyl, ethyl or isopropyl and the non-toxic, pharmaceutically acceptable acid salts with both organic and inorganic acids.
[0029] The most preferred compounds are 2-(4-thiazolyl)benzimidazole, methyl-(butylcarbamoyl)-2-benzimidazolecarbamate and 2-methoxycarbonylamino-benzimidazole and the compounds wherein Y is chloro and X is hydrogen.
[0030] The benzimidazole compounds also include prodrugs. "Prodrugs" are considered to be any covalently bonded carriers which release the active parent drug according to the formula of the benzimidazole derivatives described above in vivo when such prodrug is administered to a mammalian subject. Prodrugs of the benzimidazole compounds are prepared by modifying functional groups present in the compounds in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compounds. Prodrugs include compounds wherein hydroxy, amine, or sulfhydryl groups are bonded to any group that, when administered to a mammalian subject, cleaves to form a free hydroxyl, amino, or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, or benzoate derivatives of alcohol and amine functional groups in the benzimidazole derivatives; phosphate esters, dimethylglycine esters, aminoalkylbenzyl esters, aminoalkyl esters and carboxyalkyl esters of alcohol and phenol functional groups in the benzimidazole derivatives; and the like.
[0031] The pharmaceutically acceptable salts of the benzimidazole derivatives include the conventional non-toxic salts or the quaternary ammonium salts of the benzimidazole derivatives formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like.
[0032] The pharmaceutically acceptable salts of the present invention are synthesized from the benzimidazole derivatives which contain a basic or acidic moiety by conventional chemical methods. Generally, such salts are prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, the disclosure of which is hereby incorporated by reference. The disclosures of all of the references cited herein are hereby incorporated herein by reference in their entirety.
[0033] Synthesis
[0034] The benzimidazole derivatives are prepared in a number of ways well known to one skilled in the art of organic synthesis. The benzimidazole derivatives are synthesized using the methods described below, together with synthetic methods known in the art of synthetic organic chemistry, or variations thereon as appreciated by those skilled in the art. Preferred methods include but are not limited to those methods described below. Each of the references cited below are hereby incorporated herein by reference.
[0035] These compounds are prepared according to the method described in U.S. Pat. No. 3,738,995 issued to Adams et al, Jun. 12, 1973. The thiazolyl derivatives are prepared according to the method described in Brown et al., J. Am. Chem. Soc., 83 1764 (1961) and Grenda et al., J. Org. Chem., 30, 259 (1965).
[0036] C. Dosage and Dosage Delivery Forms
[0037] The type of compound and the carrier and the amount will vary widely depending on the species of the warm blooded animal or human, body weight, and tumor being treated. The dosage administered will vary depending upon known factors, such as the pharmacodynamic characteristics of the particular agent and its mode and route of administration; the age, health and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment; the frequency of treatment; and the effect desired.
[0038] The benzimidazole is preferably micronized or powdered so that it is more easily dispersed and solubilized by the body. Processes for grinding or pulverizing drugs are well known in the art. For example, a hammer mill or similar milling device are used. The preferred particle size is less than about 100μ and preferably less than 50μ.
[0039] The dosage administered will vary depending upon known factors such as the pharmacodynamic characteristics of the particular active ingredient, and its mode and route of administration; age, sex, health, metabolic rate, absorptive efficiency and/or weight of the recipient; nature and extent of symptoms; kind of concurrent treatment, frequency of treatment; and the effect desired.
[0040] A "tumor growth inhibiting amount" of the benzimidazole derivatives is that amount which is effective to inhibit or slow the growth of a tumor.
[0041] Dosage forms (compositions) suitable for internal administration contain from about 1.0 milligram to about 5000 milligrams of active ingredient per unit. In these pharmaceutical compositions, the active ingredient will ordinarily be present in an amount of about 0.5-95% by weight based on the total weight of the composition. Based on the body weight of the patent, the dosage may be administered in one or more doses several times per day or per week. Multiple dosage units may be required to achieve a therapeutically effective amount. For example, if the dosage form is 1000 mg, and the patient weighs 40 kg, one pill will provide a dose of 25 mg per kg for that patient. It will provide a dose of only 12.5 mg/kg for a 80 kg patient.
[0042] The compounds have shown dose responsiveness in vivo against viruses and cancers in mice at 500 mg/kg, 2500 mg/kg, 3500 mg/kg, 4000 mg/kg, 5000 mg/kg and 6000 mg/kg. Generally a dosage effective in mice translates to about {fraction (1/12)}of the dosage required in humans. By way of general guidance, for humans a dosage of as little as about 30 milligrams (mg) per kilogram (kg) of body weight and up to about 10000 mg per kg of body weight is suitable. Preferably from 50 mg/kg to about 5000 mg/kg of body weight is used. Most preferably the doses are between 100 mg/kg to about 3000 mg/kg of body weight. However, a dosage of between about 2 milligrams (mg) per kilogram (kg) of body weight to about 400 mg per kg of body weight is also suitable for some indications.
[0043] Intravenously, the most preferred doses may range from about 1 to about 1000 mg/kg/minute during a constant rate infusion. Benzimidazole derivatives may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three, or four times daily. The benzimidazole derivatives are given in one or more doses on a daily basis or from one to three times a week.
[0044] The benzimidazole derivatives may also be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittant throughout the dosage regimen.
[0045] Generally, the dosage in man is lower than for small warm blooded mammals such as mice. A dosage unit may comprise a single compound or mixtures thereof with other compounds or other cancer inhibiting compounds or tumor growth inhibiting compounds or anti-viral compounds. The dosage unit can also comprise diluents, extenders, carriers and the like. The unit may be in solid or gel form such as pills, tablets, capsules and the like or in liquid form suitable for oral, rectal, topical, intravenous injection or parenteral administration or injection into or around the tumor.
[0046] The benzimidazole derivatives are typically mixed with a pharmaceutically acceptable carrier. This carrier can be a solid or liquid and the type is generally chosen based on the type of administration being used. The active agent can be coadministered in the form of a tablet or capsule, as an agglomerated powder or in a liquid form. Examples of suitable solid carriers include lactose, sucrose, gelatin and agar. Capsule or tablets are easily formulated and can be made easy to swallow or chew; other solid forms include granules, and bulk powders. Tablets may contain suitable binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents. Examples of suitable liquid dosage forms include solutions or suspensions in water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and/or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules. Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners, and melting agents. Oral dosage forms optionally contain flavorants and coloring agents. Parenteral and intravenous forms would also include minerals and other materials to make them compatible with the type of injection or delivery system chosen.
[0047] The benzimidazole derivatives of this invention are administered as treatment for cancer and viral infections, including retroviral, by any means that produces contact of the active agent with the agent's site of action in the body. The antitumor compounds (active ingredients) of this invention are administered to inhibit tumors by any means that produces contact of the active ingredient with the agent's site of action in the body of a mammal. They can be administered by any conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutic agents. They can be administered alone, but generally are administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.
[0048] The benzimidazole derivatives are administered in oral dosage forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. The benzimidazole derivatives may also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.
[0049] In the methods of the present invention, the compounds herein described in detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, excipients, or carriers (collectively referred to herein as a pharmaceutically acceptable carrier or carrier materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
[0050] For instance, for oral administration in the dosage unit form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like.
[0051] For oral administration in liquid dosage form, the oral drug components are combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents, and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.
[0052] The benzimidazole derivatives can also be administered in the form of liposome delivery systems, such as small unilamallar vesicles, large unilamallar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.
[0053] Benzimidazole derivatives may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxylpropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates, and crosslinked or amphipathic block copolymers of hydrogels.
[0054] The active ingredient can be administered orally in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions. It can also be administered parentally, in sterile liquid dosage forms.
[0055] Gelatin capsules may contain the active ingredient and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.
[0056] Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance. In general, water, a suitable oil, saline, aqueous dextrose (glucose), and related sugar solutions and glycols such as propylene glycol or polyethylene glycols are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain a water soluble salt of the active ingredient, suitable stabilizing agents, and if necessary, buffer substances. Antioxidizing agents such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or combined, are suitable stabilizing agents. Also used are citric acid and its salts and sodium EDTA. In addition, parenteral solutions can contain preservatives, such as benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol. Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field.
[0057] Useful pharmaceutical dosage forms for administration of the compounds of this invention are illustrated as follows:
[0058] Capsules
[0059] A large number of unit capsules are prepared by filling standard two-piece hard gelatin capsules each with 100 to 500 milligrams of powdered active ingredient, 5-150 milligrams of lactose, 5-50 milligrams of cellulose, and 6 milligrams magnesium stearate.
[0060] Soft Gelatin Capsules
[0061] A mixture of active ingredient in a digestible oil such as soybean oil, cottonseed oil or olive oil is prepared and injected by means of a positive displacement pump into gelatin to form soft gelatin capsules containing 100-500 milligrams of the active ingredient. The capsules are washed and dried.
[0062] Tablets
[0063] A large number of tablets are prepared by conventional procedures so that the dosage unit was 100-500 milligrams of active ingredient, 0.2 milligrams of colloidal silicon dioxide, 5 milligrams of magnesium stearate, 50-275 milligrams of microcrystalline cellulose, 11 milligrams of starch and 98.8 milligrams of lactose. Appropriate coatings may be applied to increase palatability or delay absorption.
[0064] Injectable
[0065] A parenteral composition suitable for administration by injection is prepared by stirring 1.5% by weight of active ingredient in 10% by volume propylene glycol and water. The solution is made isotonic with sodium chloride and sterilized.
[0066] Suspension
[0067] An aqueous suspension is prepared for oral administration so that each 5 ml contain 100 mg of finely divided active ingredient, 200 mg of sodium carboxymethyl cellulose, 5 mg of sodium benzoate, 1.0 g of sorbitol solution, U.S.P., and 0.025 ml of vanillin.
[0068] The present invention also includes pharmaceutical kits useful, for example, for the treatment of HIV infection, which comprise one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a benzimidazole derivative. Such kits may further include, if desired, one or more of various conventional pharmaceutical kit components, such as, for example, containers with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent to those skilled in the art. Printed instructions, either as inserts or as labels, indicating quantities of the components to be administered, guidelines for administration, and/or guidelines for mixing the components, may also be included in the kit. In the present disclosure it should be understood that the specified materials and conditions are important in practicing the invention but that unspecified materials and conditions are not excluded so long as they do not prevent the benefits of the invention from being realized.
[0069] The following examples are illustrative and are not meant to be limiting to the invention.
[0070] Colon, Breast and Lung Tumor Cells Test
[0071] The following cell culture tests were performed to test the toxicity of the benzimidazole compounds on colon, breast and lung human tumor cells. The viability of the cells were tested by looking at MTT (3-[4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide) reduction. MTT assay is a well known measure of cell viability.
[0072] The colon tumor cells (HT29 from American Type Culture Collection (ATCC) and the breast cells (MX1 from cell lines from ATCC) were cultured in Eagle's Miminal Essential Medium with 10% fetal bovine serum. The lung tumor cells (A549 from ATCC cell lines) were cultured in Ham's F12 medium with 10% fetal bovine serum.
[0073] The tumor cells were passaged and seeded into culture flasks at the desired cell densities. The culture medium was decanted and the cell sheets were washed twice with phosphate buffered saline (PBS). The cells were trypsinized and triturated prior to seeding the flasks. Unless otherwise indicated the cultures were incubated at 37�1� C. in a humidified atmosphere of 5�1% carbon dioxide in air. The cultures were incubated until they were 50-80% confluent.
[0074] The cells were subcultured when the flasks were subconfluent. The medium was aspirated from the flasks and the cell sheets rinsed twice with PBS. Next, the Trypsin Solution was added to each flask to cover the cell sheet. The Trypsin Solution was removed after 30-60 seconds and the flasks were incubated at room temperature for two to six minutes. When 90% of the cells became dislodged, growth medium was added. The cells were removed by trituration and transferred to a sterile centrifuge tube. The concentration of cells in the suspension was determined, and an appropriate dilution was made to obtain a density of 5000 cells/ml. The cells were subcultured into the designated wells of the 96-well bioassay plates (200 microliter cell suspension per well), PBS was added to all the remaining wells to maintain humidity. The plates were then incubated overnight before test article treatment.
[0075] Each dose of test article was tested by treating quadruplicate wells of cultures with 100 microliter of each dilution. Those wells designated as solvent controls received an additional 100 microliter of methanol control; negative controls wells received an additional 100 microliters of treatment medium. PBS was added to the remaining wells not treated with test article or medium. The plates were then incubated for approximately 5 days.
[0076] At the end of the 5 day incubation, each dose group was examined microscopically to assess toxicity. A 0.5 mg/ml dilution of MTT was made in treatment medium, and the dilution was filtered through a 0.45 micrometer filter to remove undissolved crystals. The medium was decanted from the wells of the bioassy plates. Immediately thereafter, 2000 microliter of the filtered MTT solution was added to all test wells except for the two untreated blank test wells. The two blank wells received 200 microliters of treatment medium. The plates were returned to the incubator for about 3 hours. After incubation, the MTT containing medium was decanted. Excess medium was added to each well and the plates were shaken at room temperature for about 2 hours.
[0077] The absorbance at 550 mn (OD550) of each well was measured with a Molecular Devices (Menlo Park, Calif.) VMax plate reader.
[0078] The mean OD550 of the solvent control wells and that of each test article dilution, and that of each of the blank wells and the positive control were calculated. The mean OD550 of the blank wells was subtracted from the mean of the solvent control wells, and test article wells, respectively to give the corresponding mean OD550.
[0079] Dose response curves were prepared as semi-log plots with % of control on the ordinate (linear) and the test article concentration on the abscissa (logarithmic). The EC50 was interpolated from the plots for each test article.
[0080] For the test articles administered in methanol, separate responses were prepared to correct for the methanol data.3
[0081] Adriamycin was used as a positive control. In all cases, it was more toxic than any of the test materials by one or two logs. Adriamycin is one of the more potent agents in current use and one with significant side effects. The peak plasma concentration of other, quite effective chemotherapeutic agents may be 10 to 50 times higher than that of Adriamycin.
[0082] The EC50 is the concentration at which one half of the cells are killed.
[0083] In normal healthy cells, the following results were obtained. As is evident, the benomyl and carbendazim were much less toxic to normal healthy cells than adriamycin.
[0084] In a related study using lung tumor cells (A-549) breast tumor cells (MCF-7) and colon tumor cells (HT-29), thiabendazole effectively killed these cells. Table 3 summarizes the results
[0085] These experiments show that these compositions are effective in killing tumor cells of the breast, colon and lung type.
[0086] Carbendazim has shown broad-scale efficacy against multiple cancer types both in vitro and in vivo. The cancers tested include colon, lung, breast, prostate, pancreatic, leukemia, melanoma, neuroblastoma, ovarian, neck and head, and brain. Also multiple cell lines were tested in almost instances.
[0087] The initial efficacy is comparable to existing best available drugs. But with carbendazim, the tumors do not recur or reappear as happens with Cytoxan and Taxol, which otherwise are quite good against breast cancer. Similarly, pancreatic cancer does not appear to come back as often happens with Gemcitabine treatment.
[0088] Carbendazim is particularly good in mouse melanoma in mice, which many people believe is the best predictive model for efficacy in humans. It has shown outstanding broad and good results in the human tissue cloning test. This is an in vitro test on conventionally treated and recently excised human tumors.
[0089] Carbendazim is equally effective against p53 deficient/defective cell lines, unlike most existing drugs. It appears that carbendazimn induces apoptosis in cancer cells at sub-lethal concentrations to normal cells.
[0090] Other Benefits of Carbendazim are:
[0091] Its oral LD50 in mice is quite high (11,000 mg/kg), a low overall toxicity unlike most cancer drugs. For perspective the LD50 of table salt is 3750 mg/kg.
[0092] It is effective in cancers that form tumors and those that do not, e.g. both carcinomas and sarcomas.
[0093] The results of these studies are provided in more detail below.
[0094] Some of the pharmacological effects of carbendazim were demonstrated by studying its ability to induce apoptosis in cancer cells, studying its effect on p53-abnornal cell lines and determining during which cellular life cycle phase carbendazim exerts its effects.
[0095] Apoptosis Study:
[0096] Apoptosis is a specific type of cell death which differs from necrosis and is characterized by specific morphological, biochemical and micellular cell changes. Abnormalities in p53 expression are generally inked with the prevention of apoptosis and p53 abnormalities are common in human tumors which are resistant to conventional cytotoxic agents.
[0097] Summary of test results: The extent of apoptosis is measured in human tumor cell lines after treatment for 1, 2, 3 and 4 days with carbendazim. At each of these time points, the cells are harvested and assayed using the terminal deoxynucleotidyl transferase (TdT) assays. Both microscopy and flow cytometry were used for the TdT assay.
[0098] In MCF7, HT29, B16 and SK-MES cell lines, there was a concentration dependent effect on cell growth. In most cases at concentrations of carbendazim greater than 1 μg/ml, the growth rate was significantly slower than in the untreated samples. After completion of the studies on the MCF7 and HT29 cell lines, it was evident that the 0.1 μg/ml concentration had little effect on either apoptosis or cell growth. Therefore subsequent assays were with 5 μg/ml instead of 0.1 μg/ml.
[0099] The growth of MCF7 (breast cancer) cells was not significantly affected by carbendazim below 10 μg/ml as shown by either method. However, at 10 μg/ml the increase in apoptosis was evident at days 3 and 4. The increase in apoptosis was low, less than 10% at the high concentration. After normalization for the cell growth, carbendazim at 10 μg/ml had an effect at day 4.
[0100] The growth of HT 29 (colon) cancer cells was not largely slowed down by carbendazim below 10 μg/ml. Concentration-dependent increase in apoptosis was observed at days 3 and 4, reaching >25% apoptosis in the presence of 10 μg/ml carbendazim. After normalization for the cell growth rates, the concentration-response effect was seen at day 4.
[0101] At all concentrations, carbendazim affected the growth rates in B16 murine melanoma cell line. Some concentration dependent effects on apoptosis were seen at days 1-4 by TdT microscopy and days 2-4 by flow cytometry. The concentration effects at days 1-4 were much more evident after normalization of the apoptosis for the growth rates.
[0102] The growth of SK-MES cells was slowed done by all concentrations of carbendazim past day 1. The percent of apoptosis showed a concentration-response effect at days 1-4 by microscopic TdT assay and by flow cytometry at days 3 and 4. A normalized graph showed a concentration response effect of the compound on apoptosis at all days.
[0103] Conclusion: At concentrations less than 10 μg/ml, the greatest response to carbendazim was seen in the SK-MES lines (lung), followed by B16 (murine melanoma), HT29 (colon), and MCF7 (breast) cells. The HT29, SK-MES and B16 cell lines express abnormal p53. Accordingly carbendazim can induce apoptosis in p53 abnormal cell lines.
[0104] Selectivity in Killing p53 Abnormal Cell Lines
[0105] Carbendazim provides in vivo activity against HT29 tumor cells which express abnormal p53.
[0106] Summary of test results: Pairs of tumors of the same type were chosen, one expressing normal p53 and the other abnormal p53. Breast lines used were MCF7 for the normal p53, V4B, a MCF7 cells transformed with an empty vector, and VM4K, an MCF7 cells transfected with a vector encoding abnormal p53. In the colorectal cancer model HCT116 with a normal p53 was used and DLD-1 was used as the abnormal p53 cell line. The tumor cells were grown the presence and absence of carbendazim for 7 days. Cell growth rates were determined in each group by counting the cell numbers daily using a Coulter counter.
[0107] In the breast model at day 7, using 1 μg/ml of carbendazim, cell counts in all three cell lines were between 50% and 60% of the control. At concentrations of 5 and 10 μg/ml the MCF7 cell counts were 45% and 36% of the control. The V4B and VM4K cell lines were less than 10% of the respective controls. The data from the VM4K cell line would also indicate the carbendazim is again selectively killing the p53 abnormal cells, however, there was also a marked decrease in the cell numbers in the V4B line carrying only the empty vector. In light of the controversy regarding the p53 status in MCF7 cells, which some researchers claim to express abnormal p53, these differences may be more difficult to interpret.
[0108] At day 7 in the colorectal model, at 1 μg/ml concentration carbendazim cell counts in the DLD1 line (abnormal p53) were only 34% of the control. The HCT 116 cell line, (normal p53) showed cell counts that were 78% of the control suggesting that carbendazim may selectively kill p53 abnormal cells. At carbendazim concentrations of 5 and 10 μg/ml, cell counts were less than 10% of the control in both the DLD-1 and HCT116 lines indicating that at the larger concentration, the drug was equally toxic to cell lines containing abnormal and normal p53.
[0109] Conclusion: At higher concentrations, 5 and 10 μg/ml, the drug was equally toxic to cell lines containing abnormal p53 and normal p53 in both colon and breast cancers.
[0110] A dose response effect of carbendazim in a human tumor cloning forming units study (HTCU) is summarized below. This is an in vitro test of treatments on conventionally treated, then recently excised human tumors. It is an important study because carbendazim is showing effectiveness against cell lines which have survived conventional treatment and which themselves have not had undergone too many passages (due to recent excision). This is significant since long living cell lines undergo changes (passages), some of which may affect their resistance to some drugs, and hemotherapeutic agent resistant cells can be formed. The data show the activity as the tumors tested that had <50% survival (a high number is desirable).
[0111] Solid tumors removed by patients are minced into 2 to 5 mm fragments and immediately placed in McCoy's Medium 5A plus 10% heat inactivated newborn calf serum plus 1% penicillin/streptomycin. Within 4 hours, these solid tumors are mechanically disassociated with scissors, forced through No. 100 stainless steel mesh, through 25 gauge needles, and then washed with McCoy's medium as described above. Ascitic, pleural, pericardial fluids and bone marrow are obtained by standard techniques. The fluid or marrow is placed in sterile containers containing 10 units of preservative free heparin per ml. of malignant fluid or marrow. After centrifugation at 150�g for 10 minutes, the cells are harvested and washed with McCoy's medium plus 10% heat inactivated calf serum. The viability of cell suspensions is determined on a hemocytometer with trypan blue.
[0112] Cells to be cloned are suspended in 0.3% agar in enriched CMRL1066 supplemented with 15% heat inactivated horse serum, penicillin (100 units/ml), streptomycin (2 mg/ml), glutamine (2 mM), insulin (3 units/ml), asparagine (0.6 mg/ml), and HEPES buffer (2 mM). For the continuous exposure test each compound is added to the above mixture. Cells are placed in 35 mm petri dishes in a top layer of agar over an underlayer of agar to prevent growth of fibroblasts. Three plates are prepared for each data point. The plates are placed in a 37� C. incubator, and are removed on day 14 for counting of the number of colonies in each plate. The number of colonies (defined as 50 cells) formed in the 3 compound treated plates is compared to the number of colonies formed in the 3 control plates, and the percent colonies surviving at the concentration of compound can be estimated. Three positive control plates are used to determine survival rate. Orthosodium vanadate at 200 μg/ml is used as the positive control. If there is <30% colonies in the positive control when compared to the untreated control, the test is evaluated.
[0113] (4-thiazolyl)-1H-benzimidazole shows efficacy in the Human Tumor Colony Forming Units test described above when tested using continuous exposure of the cells to the (4-thiazolyl)-1H-benzimidazole.
[0114] The following table summarizes these results showing positive results on a number of cancer types:
[0115] The dose response effect of carbendazim in mice infected with various cancer types was studied in standard screening tests.
[0116] The results of these studies conducted to study the dosage response of carbendazim on various cancer types are summarized below. These results are representative of a number of tests in which a known chemotherapeutic agent is used as the control so that the efficacy of the carbendazim can be compared to it. Efficacy of carbendazim has been shown in the following cancers:
[0117] In a Prostate cancer model doses of 4000, 5000 and 6000 (mg/kg given once weekly) were as effective as Mitoxantrone at 70 days. It was better than Cytoxan at these doses of 4000, 5000 and 6000 (mg/kg given twice weekly) through 40 days. The tables show tumor weight in mg for each dose. The carbendazim was given once a week (p.o) and the Mitoxantrone was given by i.v. (q.d.x5).
[0118] Both the Cytoxan and the carbendazim were given p.o. twice a week
[0119] In the Colon—HT29 mouse model carbendazim at doses of 4000, 5000 and 6000 (mg/kg given twice weekly) it is better than Cytoxan in this model. At a dose of 3000 (mg/kg given twice weekly) it is also better than Cytoxan.
[0120] The carbendazim and Cytoxan were given twice weekly.
[0121] In the Breast—MX-1 model, carbendazim at 4000, 5000, and 6000 (mg/kg, twice weekly) was dose responsive in slowing the growth of the tumor and was better than Cytoxan. It is also very effective in MCF-7L breast line. At a dose of 3000, (mg/kg given twice weekly) it was equivalent to Navelbine.
[0122] The Cytoxan and the carbendazim were given twice weekly, p.o.
[0123] When the tumor was shrunk with Taxol first and then the carbendazim therapy started when the tumor began to grow again on day 130, the carbendazim treatment was begun and the tumor subsequently shrunk to zero. The Navelbine was given 1.6 mg./kg, qdx5, i.p; the Taxol was given 16 mg/kg, qdx5, i.p; and the carbendazim was dosed twice weekly, p.o. The carbendazim treatment was better than Navelbine and as good as Taxol in this study.
[0124] In a third study carbendazim was tested in mice MX-1 model with the mice receiving estrogen tablets. Estrogen accelerates the growth of the breast cancer. At doses of 2000, 4000 and 6000 (mg/kg given once weekly) it was better than both Navelbine (1.6 mg/kg, qdx5, i.p.) and Taxol (16 mg/kg, qdx5, i.p.) in this faster growing MX-1 cancer.
[0125] In the Pancreas (Mia-PaCa) model, carbendazim at 3000 and 4000 (mg/kg, twice weekly) is as good as or better than gemcitabine. At 2000 mg/kg the carbendazim was not as effective after 21 days. Gemcitabine was given on days 1,4,7 and 10 i.p.; the carbendazim was given p.o. twice weekly.
[0126] In the Panc-01 model for pancreatic cancer carbendazim at a dose of 5000 mg/kg was better than Gemcitabine at 32 days. The Gemcitabine is given i.p., q3dx4 and the carbendazim is given p.o. twice weekly to the end.
[0127] In the Neuroblastoma (SK-N-MC) model a decrease in tumor growth at 5000(mg/kg) is shown and there is favorable activity early in the study compared to topotecan. It is expected, based on learning from the other studies, that a lower dose would be effective. The topotecan is given i.p. qdx5 and the carbendazim is given p/.o. twice weekly.
[0128] In the Rhabdomysarcoma model, a childhood cancer, at day 29 carbendazim at doses of 3000, 4000 and 5000 (mg/kg given once weekly, p.o.) is quite effective against this tumor with no toxicity in the 3000 (mg/kg) group. Topotecan (i.p., qdx5) is quite active against this model.
[0129] In the Lung cancer model MV522 all groups received Taxol and then received treatments shown starting 10 days later. At doses of 2000, 3000, 4000 and 5000 (mg/kg given twice weekly, p.o.) tumor growth was suppressed in a dose response and the tumor was shrunk in the 5000 mg/kg group. Day 1 is the start of the treatment with carbendazim.
[0130] In a Medulloblastoma model (IMR32) a dosage dependent effect is seen with carbendazim at doses of 5000, 4000, and 3000 (mg/kg, p.o. twice weekly). It is compared with Topotecan (i.p. qdx5).
[0131] In the Murine Melanoma—B16 mice model doses of 4000, 5000 and 6000 (mg/kg) were at least equal in tumor suppression compared to cytoxan at 30-60 days. Since this tumor is a liquid tumor, there is no change in tumor weight. The results are summarized in Table 1 below.
[0132] In the P-388 model for leukemia there was a dose responsive effect and the results were good. See Tables 2 and 3.
[0133] The data in Tables 1, 2 and 3 are reported in T/C which is interpreted using the following scale:
[0134] long term survivors for P388 is >30 days, for B16 it is >60 days. The NCI Measure of Success is T/C=125.
[0135] This study was performed in black mice injected i.p. with B-16 mouse melanoma which many researchers believe is the most predictive model for efficacy in humans. Carbendazim was equally effective as Cytoxan in this model at treating B-16. The activity is dose responsive. Death is the end point in this, model.
[0138] Accordingly, carbendazim retards the growth of both solid and liquid tumors in vivo.
[0139] These studies confirm previous in vivo studies in which carbendazim was tested in various in vivo models for different types of cancer.
[0140] Additional in vivo cancer studies are presented in a tabular form below. The data is presented in the following format—dose regimen in parenthesis, dosage in mg/kg: tumor growth suppression (tg supp.), toxic deaths/number of test subjects (deaths), % weight change (wt).
[0141] Carbendazim demonstrated the ability to reduce tumor growth in mice models for breast, lung, colon, murine melanoma and leukemia. The data are summarized in Table 5 and Table 6 below.
[0142] In the same test 2-(4-thiazolyl)-1H-benzimidazole showed no activity against MXI breast cancer tumors implanted subcutaneously under the mice skin.
[0143] In the same test 2-(4-thiazolyl)-1H-benzimidazole showed no activity against P388.
[0144] In the same test 2-(4-thiazolyl)-1H-benzimidazole showed no activity against B16.
[0145] The in vivo and in vitro data support the assertion that carbendazim has broad scale efficacy against multiple cancer types.
[0146] Initial efficacy of carbendazim appears to be comparable to the best available drugs used for the treatment of any particular cancer type. Furthermore, with continuous treatment, breast cancers do not come back as usually happens with cytoxan and taxol in breast cancer or gemcitabine in pancreatic cancer.
[0147] It is equally effective against p53 deficient/defective cell lines, unlike most existing cancer treatment drugs.
[0148] It is excellent in B16 mouse melanoma, which is believed by many people to be the best predictive model for efficacy in humans.
[0149] These same benzimidazole derivatives are effective against viruses including HIV, influenza, rhinoviruses and herpes viruses. The benzimidazole derivatives can be used alone or in combination with other fungicides.
[0150] The following examples illustrate the effectiveness of thiabendazole, 2-(4-thiazolyl)-1H-benzimidazole, against HIV and the benzimidazoles derivatives against a number of viruses.
[0151] The results of these HIV studies are summarized in more detail below:
[0152] Thiabendazole is effective at totally suppressing virus production in chronically infected cells. The extra cellular viral count goes effectively to zero or non-detectable levels. Thiabendazole does not kill the chronically infected cells though it does reduce the rate of cell proliferation at active concentrations. Thiabendazole does not affect CD4 expression in uninfected cells. At effective concentrations thiabendazole slows but does not alter the normal cellular RNA or protein synthesis of either infected or non-infected cells. Thiabendazole is effective in a variety of chronically infected cell types (this effect is not cell type specific.)
[0153] Thiabendazole is effective against a variety of HIV virus strains. (Not virus strain specific—although some variance by strain is observed; SK-1>IIIB>RF) Also thiabendazole is not effective on SIV in vitro or in vivo.
[0154] After 20 months no resistant virus strains to thiabendazole have developed in tests designed to do so. Resistance develops in six months or less in this test for existing HIV drugs with resistance strains for protease inhibitors developing in about 3-4 months.
[0155] Thiabendazole does not adversely affect the activity of existing HIV drugs, AZT, 3TC, ddC, dd1 or protease inhibitors (saquinavir and indinavir) in acutely infected cells, nor do any of these existing drugs interfere with the efficacy of thiabendazole in chronically infected cells. It is used in combination with these drugs. Thiabendazole is also effective against protease inhibitor resistant viruses.
[0156] Thiabendazole confers temporary suppression of viral production from 4 to 80 days after treatment stops. This is unique and a useful feature whenever one has problems with compliance.
[0157] The results of these studies are summarized in detail below:
[0158] Thiabendazole was tested in chronically infected HIV virus. These cell populations contain integrated copies of the HIV genome and constituitively produce HIV at relatively high levels (CEM-SK1, U937-SK1 and H9-SK1 from Frederick Research Center, Maryland) or are latently infected and only produce virus after stimulation with phorbol esters, tumor necrosis factor or IL6 (U1 and ACH2). Virus production was reduced in all cell lines tested and thiabendazole did not stimulate virus production from the latently infected cells. Reductions in virus production were observed when quantifying supernatant reverse transcriptase activity, supernatant p24 as well as intracellular p24, indicating the compound inhibits virus production at a step of replication prior to production of intracellular proteins.
[0159] Quantification of the infectivity of virions produced from the infected cells demonstrates reductions in the number of infectious virions in parallel with reductions in supernatant RT or p24, indicating the compound reduces the amount of virus produced, but not the quality of the virions. Inhibition of virus production from the chronically infected cells was observed at concentrations which were nontoxic to the target class. Thiabendazole inhibited virus production at concentrations greater than 1-10 μg/ml.
[0160] Toxicity to the chronically infected cells was similar to that observed with the uninfected cells. Evaluation of thiabendazole on chronically infected cells was performed by evaluation of thymidine (DNA), uridine (RNA) and leucine (protein) incorporation into cellular macromolecules. Inhibition of cellular macromolecule synthesis paralleled the toxicity of the compound as would be expected and did not occur at lower nontoxic concentrations found to inhibit virus production from the chronically infected cells.
[0161] After 28 days of treatment on chronically infected cells, the toxicity of the compound to the target cells appeared similar in both uninfected and chronically infected cells. The compound does not preferentially kill HIV-infected cells. Reductions in the level of virus production were stable and were observed at concentration greater than 10 μg/ml for thiabendazole.
[0162] These results suggest thiabendazole can quickly reduce the level of virus production from cell populations chronically infected with HIV-1 and the antiviral effect is maintained with prolonged compound exposure. This reduction of virus production occurs at concentrations which are nontoxic to the host cell and which have no effect on the synthesis of cellular DNA, RNA and protein.
[0163] Chronically infected HIV cells were cultured in the presence of thiabendazole at 1 μg/ml for the first month, 5 μg/ml for the second month, 10 μg/ml for the third month, 20 and 40 μg/ml for the fourth month and 80 μg/ml for the fifth and sixth months. At the end of each month, the cells were evaluated for virus production compared to chronically infected cells not treated with the compound. For each of the six months of treatment experience, no change in the antiviral effect of the compound was noticed and the toxicity of the compound remains identical. Thiabendazole remains active against HIV and that resistance was not rapidly achieved via the selection of resistant viruses or adaptation of the cells to prevent compound induced toxicity. Virus production remains totally suppressed from cultures treated with thiabendazole at 40 and 80 μg/ml.
[0164] The present invention includes a method of treatment HIV with the benzimidazole compounds without inducing formation of benzimidazole derivative or thiabendazole resistant HIV.
[0165] Chronically infected cells which were treated with compound for prolonged periods were washed free of compound and cultured to determine if, and when, virus production would resume. Cultures in which treatment resulted in the total elimination of virus production were used in these assays. These cultures included chronically infected cells cultured in the presence of 20, 40, and 80 μg/ml of thiabendazole. Within 4 days virus production resumed from the cells cultured in the presence of the lower concentrations of thiabendazole (20 μg/ml and 4 μg/ml). Virus production resumed at the 40 μg/ml concentration of thiabendazole by day 12. At the highest concentrations virus production was observed at approximately day 70.
[0166] The present invention includes a method of treatment HIV with the benzimidazole derivative or thiabendazole and delaying the reappearance of HIV in plasma following initial treatment of HIV with an antiviral agent or thiabendazole.
[0167] Cells which were pretreated with thiabendazole for a long period of time were washed free of compound and used as a target cell population. The cells were split into 3 populations and labeled Group 1, 2 or 3. Group 1 was treated with the compound for 24 hours (at the same concentration used in the prolonged treatment phase), washed free of compound and cultured in the presence of infectious virus and fresh compound. Group 2 was pretreated for 24 hours, washed free of compound and cultured in the presence of infectious virus only. Group 3 was cultured for both the pretreatment and the infection phases in fresh medium only (no virus or compound). Virus production from the cell populations was identical irrespective of the culture conditions. These results indicate that the chronically infected cells treated for prolonged periods were not super-infected with HIV.
[0168] Chronic HIV-1 infected cells U1 were derived from an acute HIV-1 infection of the promonocytic cell line, U937. The chronic HIV-1 infected cells, ACH-2 were derived from an acute HIV-1 infection of the T cell line, A3.01.
[0169] These cells were cultured in medium and the phorbol ester, PMA. PMA causes the cells (both U1 and ACH-2) to be activated and not divide but it also causes the U-1 cells to differentiate. This results in fewer cells in the PMA-treated cultures than the media alone cultures. Cell viability was measured when these cell lines were treated with the test compound.
[0170] Both cell lines constituitively produce a small amount of HIV-1. ACH-2 cell lines tend to produce more HIV-1 than U1 cells as shown by p-24 ELISA. When either cell line is cultured in the presence of PMA there is an -increase in the quantity of HIV-1 produced as measured by the p-24 antigen ELISA.
[0171] In addition, the number of institute positive HIV mRNA expressing cells per microscopic field is measured. Comparisons can be made from these numbers since the same number of cells were adhered to the glass slides for each drug concentration (10�106 cells/ml).
[0172] These cells were treated with test samples. Thiabendazole at 60 μg/ml suppressed replication in the HIV monocytes by 74% and the T-cell HIV replication was increased by 26%. The positive control was interferon which suppressed HIV monocytes replication by 80%. AZT showed no activity in this model.
[0173] 2-(Methoxycarbonylamino)benzimidazole suppressed replication in the HIV monocytes by 9% and the T-cell HIV replication was increased by 44%. The positive control was interferon which suppressed HIV monocytes replication by 80% and suppressed T-cell HIV replication by 60%.
[0174] In an in vitro acute model for HIV 2-(methoxycarbonylamino)benzimidazole inhibited viral replication by 100% at 4 μg/ml and AZT inhibited viral replication by 98% at 1 μg/ml. 2-(4-thiazolyl)-1H-benzimidazole inhibited viral replication by 98% at 60 μg/ml.
[0175] The therapeutic index (TI), the ratio of the toxic dose of drug to efficacious dose of drug for 2-(4-thiazolyl)-1H-benzimidazole is 2.8 versus 12, 500 for AZT. The TI for 2-(methoxycarbonylamino)benzimidazole is 1.8.
[0176] In an in vivo herpes screening test of 2-(4-thiazolyl)-1H-benzimidazole at a dose of 200 mg/kg dose, 10% of the mice survived with a 10.4 mean death date; at 100 mg/kg dose 50% of the mice survived with a 9.2 mean death date. The positive control was acyclovir at 75 mg/kg dose; 60% of the mice survived with a mean death date of 17.2 days. In the same test 2-(methoxycarbonylamino)benzimidazole showed no activity.
[0177] Both 2-(4-thiazolyl)-1H-benzimidazole and 2-(methoxycarbonylamino) benzimidazole were tested in an in vitro influenza model and showed no activity.
[0178] In an in vivo model for influenza 2-(4-thiazolyl)-1H-benzimidazole was tested at 200 mg/kg, 67% of the mice survived with a mean death date of 8 days; at 100 mg/kg, 62% survived with a mean death date of 8.7 days. The positive control was amantadine (75 mg/kg) with 100% of the mice surviving for 21 days. 2-(Methoxycarbonylamino) benzimidazole was not active in the same test.
[0179] Both 2-(4-thiazolyl)-1H-benzimidazole and 2-(methoxycarbonylamino) benzimidazole were tested in an in vitro herpes model and showed no activity.
[0180] Both 2-(4-thiazolyl)-1H-benzimidazole and 2-(methoxycarbonylamino) benzimidazole were tested in an in vitro rhinovirus model and compared to A-36683. The therapeutic index (TI), the ratio of the toxic dose of drug to efficacious dose of drug, for 2-(4-thiazolyl)-1H-benzimidazole is 1-2 and for 2-(methoxycarbonylamino) benzimidazole is 1-3 versus 1000-3200 for A-36683.
[0181] The demonstrated effectiveness of the compounds of the present invention in the human breast and lung tumor xenograft models indicate that the compounds of the present invention are useful for the treatment of solid tumors in man, and, in particular, tumors of the breast and lung. This conclusion is further supported by published analyses correlating pre- clinical test results with clinical efficacy of anti-cancer agents. For example, see: Goldin and Venditti (1980) Recent Results Cancer Research 76: 176-191; Goldin et al. (1981) Eur. J. Cancer 17: 129-142; Mattem et al. (1988) Cancer and Metastasis Review 7: 263-284; Jackson et al. (1990) Cancer Investigations 8: 39-47. Based on these published analyses, the exceptional high level of antitumor activity exhibited by the presently claimed compounds provide strong evidence that the compounds claimed in present invention have therapeutic utility in the treatment of cancer in man and that they will improve the quality of life of the patient.
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Obama told ABC's Chris Cuomo on "Good Morning America" Thursday that this was "standard practice in Illinois" to try to improve bills lawmakers didn't agree with. (But that doesn't explain why it appeared to be more standard for him than some of his colleagues.)
"I worked on tons of tough bills. I was a leader on very controversial stuff," Obama said.
And he said he's glad to take the incoming fire, since it means he's a factor in the race.
"It actually beats the alternative," Obama said. "We wouldn't be doing well if people weren't confident that I could lead this country. . . . This election could be a defining moment for our politics."
Clinton, D-N.Y., would probably agree with that assessment. And she looks like she's going into the final pre-Christmas campaign days with some of her old swagger back.
"It's really picking up steam, and that's what I feel," she tells ABC's Cynthia McFadden in a "Nightline" piece that aired Wednesday night.
Clinton believes voters will ultimately look at her record: "We've gone through trying to decide, who would you rather have a beer with, and look at the results. I think we want to say, well, who would be the best president?"
And Clinton concedes that the troop surge in Iraq is having military benefits, but says it's irrelevant because Iraqi leaders aren't making political progress.
"So I don't think it matters at all. I think we need to begin to bring our troops home. In fact, I would argue that we not only need to bring them home because they should come home, but we need to bring them home because it's the only way to get the Iraqi government to focus on what it must do."
Among the Republicans, as we bid adieu to Tancredo, R-Colo., fresh evidence emerges of just how crazily tight the contest is. Romney's attacks notwithstanding, Huckabee, R-Ark., is the clear Iowa leader in the new ABC News/Washington Post poll: It's 35-27, and none of the other candidates are looking like they'll be factors on Jan. 3.
"Religion is driving the Republican presidential race in Iowa, with Mike Huckabee taking the lead on the strength of overwhelming support from evangelical voters -- and Mitt Romney falling behind over concerns about his Mormon faith," ABC Polling Director Gary Langer writes.
"Romney, for his part, holds a slight lead among the nearly eight in 10 Iowa Republicans who say his religion doesn't matter in their vote. But the remaining two in 10 say his Mormon religion makes them less likely to support him, and they overwhelmingly favor Huckabee by a large enough margin to put him in front overall."
"Republican women, particularly those who describe themselves as evangelicals and those who attend church regularly, are the primary force behind Huckabee's recent increases," Jon Cohen and Chris Cillizza write in The Washington Post.
"Women now support him over Romney by an 18-point margin; men divide their votes about evenly between the two."
And in a new national poll, "Rudy Giuliani has lost his national lead in the Republican field after a flurry of negative publicity about his personal and business activities, setting the stage for what could be the party's most competitive nomination fight in decades," John Harwood reports in writing up the new Wall Street Journal/NBC numbers.
"The results signal a dramatic shift in the nature of the Republican race: In a party with a history of rewarding established front-runners, there's no longer a front-runner of any kind."

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Who Should Go Home on 'American Idol'?
The field is down to thirteen finalists. Though last night was much better than last Wednesday night's performances, there were still more bad singers versus good singers -- in my opinion.
The field is down to thirteen finalists. Though last night was much better than last Wednesday night's performances, there were still more bad singers versus good singers -- in my opinion.
Thanks for joining us for a really great show! Check out the photos from the evening.
"Lover, lover you don't treat me no good no more", are the very popular hooky lyrics in the course of Arista Recording artist Jerrod Niemann's first ever country #1 titled "Lover, Lover".
The Music and Memories of February 28
It seems like every time you turn around it's time for another awards show. Well, here we go again. This Sunday night, it's time for Hollywood's biggest night: "The Academy Awards", otherwise known as "The Oscars". This will be two years in a row for country music to be part of "The Oscars".
Maxim Magazine recently released their list of the Hottest Women in Country Music and surprisingly Gwyneth Paltrow comes in at number one.
At 21 years old, according to Billboard, top country music earners Toby Keith, Reba, Sugarland and even the king, George Strait, didn't make as much money as Taylor Swift did in 2010.
Let's take a look at some other Grammy nominees.
On February 13, Sunday night will be the 53rd Grammy Awards. As I looked over the list of nominees, and I kept seeing this one name appear to be nominated in a few categories, and not just for country music.
In the country music industry, there are a lot of artists that sing a romantic love song at one point or another. Either the song is about loving and staying or loving and leaving.
Valentines day is coming up next week and CMT will be having a special this weekend counting down the Top 40 Greatest Love Songs. Included in the countdown will be interviews with stars such as Jason Aldean, Kenny Chesney, Lady Antebellum, Miranda Lambert, Keith Urban and Rascal Flatts. The special premieres Saturday at 8p.
Saturday night at Proctors proved to be a great one, with fantastic performances from Craig Campbell, Darryl Worley, Joe Nichols, and our great listeners in the audience.


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GQ's digital edition designed specially for your device
Only £1 an issue!
By GQ magazine 23 January 14
It's time to upgrade your life. From bespoke smartphones to the hippest hotels - from the slickest suede bomber to the starlets set for Hollywood glory, GQ has compiled a cool century of everything you will want to wear, drive, eat, hear, drink, watch and experience in 2014. Get ready for the shock of the new...


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The New Jersey shore areas are being hammered as Hurricane Sandy continues to wreak havoc along the East Coast.
Access points to and from Ocean City are all blocked due to flooding. Ocean City Patch reports that the city has declared a State of Emergency and has banned local travel.
Breakingnews.com reports a tweet from Aubrey J. Whelan regarding the Atlantic City boardwalk. According to police, the famed boardwalk's north end is collapsing.This is happening while wave heights 100 miles off the coast of New Jersey are likely to hit 30 feet or more.
According to an Associated Press report on phillyburbs.com, more than 5,000 homes and businesses had lost power as of Monday morning. Most of those buildings were in Ocean and Cumberland counties.
Governor Christie announced that the Garden State Parkway is closed in both directions from Exit 38 to the south end. Back in Atlantic City, Christie reminded people in areas likely to be hit the worst as only he can, "Don't be stupid. Get out."
Many people didn't listen elsewhere. CNN reported that rescue efforts were taking place in Oak Orchard, Delaware, a community that was under a mandatory evacuation order. The National Guard and firemen and police were rescuing people who remained Monday morning.
More than half of the 590 school districts in New Jersey closed on Monday, and 247 districts will remain closed on Tuesday.
The National Weather Service reports that a high wind warning will remain in effect until 9 a.m. on Tuesday. Winds will increase to 40 to 55 mph by 2 p.m. with gusts of 65 to 75 mph inland but 75 to 85 mph along the coasts late this afternoon.The strongest winds are expected to occur this afternoon and early evening. Wind gusts will still be 50 to 60 mph by Tuesday morning.


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This is the latest screencast I did with Adam in the team. This time it was on iSCSC and he provided a quick introduction to iSCSI, which is a technology which is part of Windows Storage Server which allows SCSI commands to be shared over the network to enable cheap and simple network storage options. In this 11min screen cast, Adam walks through iSCSI basics.
How to setup iSCSI on Windows Server 2008 [11 mins]


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Dotsie wrote:Jan - being far too nice to Pooh lately, agreeing with him all the time. A guilty conscience? Or trying to deflect suspicion?
Willem wrote:My obsession with counting and numbers isn't to be blamed on a hypothetical calculating murderous' mind - I'm a Math teacher, so I belong to a whole different league of criminals.
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And now Ashton Kutcher explains the importance of privacy, while being quoted in an international magazine.
The model-turned-TV-actor has opened up about his budding relationship with stunner Mila Kunis. And, while things may be heating up for the former "That '70s Show" co-stars, he vows that you'll be the last to hear about it.
The "Two and a Half Men" actor spoke at length about the subject with Elle magazine.
"You know, I've learned the hard way how valuable privacy is," he said. "And I've learned that there are a lot of things in your life that really benefit from being private. And relationships are one of them. And I am going to do everything in my power to have this relationship be private."
For all other details, the stud urges fans to "fill in the blanks."
Ashton also recalled the media mess resulting from his fling in 2011 with a 22-year-old, who proceeded to sell the details of their encounter to the press. His excuse for that more or less reads: When you're super famous like me, these things just kind of happen.
"You start to have suspicions about people's intentions that you're becoming close with," he said. "Because I think there's been an artificial value that's been put on fame in the world—people don't want to do something. They just want to be famous."
Read Ashton's full interview here.


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CROWD PLEASERS; Moms Read With Heart, Dads With a Firm Jaw
By JANET MASLIN
Published: May 7, 2004
Mother likes to read. She takes a special interest in American history. But she's getting tired of all those patriarchs whose every harumph has been analyzed by biographers in the throes of Founding Father Fever. Instead, Mother wonders, while Benjamin Franklin spent years in Paris admiring the wiles of Frenchwomen and studying their exact methods for putting on makeup, who was back in Philadelphia minding the store?
This is the time of year when such questions are easily answered. The publishing flurry that surrounds Mother's Day means a spate of books with conspicuously feminine points of view. Fitness, gossip, middle-aged romance, the dating woes of bright young things and anything about Jacqueline Kennedy Onassis: these are standard subjects in the parade of Mother's Day titles. They are not to be confused with the sports lore, war stories and tough-but-fair paternal advice that arrive for Father's Day.
Would it be wrong to generalize too much about who reads what, genderwise? Maybe there are men who find it interesting that as a birthday gift to one of his sisters, Benjamin Franklin decided on a spinning wheel instead of a tea table (''when I considered that the character of a good housewife was preferable to that of being only a pretty gentlewoman''). Maybe there are women whose favorite writer is Tom Clancy. But it's reasonable to assume that there is a gender gap between a book called ''The Bases Were Loaded (and So Was I)'' and anything with ''Shopaholic'' in its title.
Cokie Roberts's ''Founding Mothers'' is tailormade to the specifications of Mother's Day. Ms. Roberts has had the smart (if not brand-new) idea of collecting Revolutionary War-era stories about women who were notable, whether by accomplishment or association. Even when the facts are familiar, they take on a new aspect when viewed from a female perspective. To Esther Burr, her son Aaron was ''a little dirty noisy boy'' who was ''very sly and mischievous'' and required ''a good governor to bring him to terms.'' This woman seemed to know, as historians would later, that Aaron might be someone only a mother could love.
Ms. Roberts's book benefits not only from the sheer Mom-would-be-crazy-about-this nature of its premise but also from the author's high profile in television news. Its Father's Day counterpart, from another highly visible news figure, is ''Big Russ and Me'' by Tim Russert, an autobiography full of tough love from the author's father. Here's Buffalo-based anecdotal advice about the value of honesty, loyalty, responsibility and a firm handshake. Throw in a little baseball (it does), and the whole thing is a testament to solid values, hard work and filial devotion.
Both these books have the advantage of being reputable, a quality that heightens their seasonal, parent-friendly cachet. The same can hardly be said for ''Farewell, Jackie: A Portrait of Her Final Days'' by Edward Klein (a former editor of the New York Times Magazine and also the author of a 1989 Vanity Fair article entitled ''Jackie, Yo!''). This is the author's fifth and flimsiest effort to repackage Kennedy family history. It includes a chapter entitled ''The Pluses and Minuses of Fame.'' And it illustrates the minuses all too clearly: if you're famous enough, somebody may try to squeeze gossip out of your yoga teacher or your Italian maid.
Fatal illness gets the sob-sister treatment here, in a book that is clearly aimed at women. (When Mrs. Onassis developed non-Hodgkin's lymphoma, the reader learns, she decided that it was O.K. to start eating dessert.) But when the heartbreak has a masculine angle, soap opera won't do. In fact, heartbreak won't do. John Feinstein's ''Caddy for Life'' pointedly subjugates the sad part of its story to the life-affirming, triumphant part.
Although Bruce Edwards, the subject of Mr. Feinstein's book, has the most sports-related of all terminal illnesses (ALS, or Lou Gehrig's disease), Mr. Feinstein chooses to be manfully big-hearted rather than morbid. His is a multi-hanky book, but it is no more pessimistic -- or less bear-huggingly sentimental -- than such classic sports-tragedy dramas as ''Bang the Drum Slowly'' and ''Brian's Song.''
In telling the story of Mr. Edwards's enduring caddy-player partnership with the golf star Tom Watson, ''Caddy for Life'' accentuates the heroism of a big, lovable guy and emphasizes his rich, full life rather than his precarious medical situation. By the end of the book, Mr. Edwards has named a new dog Hope. And the final image here is a glimpse of caddy and player together, ''closer than brothers.'' On the other hand, as Mr. Watson points out, this should not be seen as a bittersweet story. ''I can honestly say it's a lot more bitter than sweet,'' Mr. Watson says.
Maybe Dad likes his sports stories a little tougher. In that case, ''The Bases Were Loaded (and So Was I)'' by Tom Callahan combines lively, swaggering sports reporting with machismo to burn. ''That was the cover that would have gagged a goat,'' Mr. Callahan writes, about the Time magazine photo of the boxer Gerry Cooney posing with Sylvester Stallone to promote ''Rocky III.'' This is not the kind of book that takes such outrages lightly.


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Confessed 'Boston Strangler' body to be exhumed
(Boston Globe) The body of Albert H. DeSalvo will be exhumed to allow for forensic testing that may answer whether he was the Boston Strangler who terrorized Greater Boston during the 1960s.
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Is he EVER going to propose?
my boyfriend and I have been together for 6 years. we have talked about marriage and kids many times before. we both want the same things in life and... Show More
Most Helpful Opinion
What is stopping you from proposing to him? So its not the 'traditional' way, but plenty of women ask the man these days. If you wanna marry him that bad then just do it. You shouldn't be talking about ultimatems yets because you haven't explored all you options. If he says no if you ask him, then I think that would be the time for ultimatems. But how do you know he is not saving up for a nice engagement ring already? Just don't be too hastey or you'll do something you'll regret. but I do suggest you asking him if he doesn't ask you soon.
Worse case scenario: he says no, you leave him. If he doesn't ask you soon your going to leave him anyway, so what will it hurt?
Hope this helped, good luck :)
What Guys Said 0
Be the first guy to share an opinion and earn 1 extra Xper Point!
What Girls Said 3
Don't talk about marriage with him anymore. Don't think about marriage with him. Just STOP!
If he truly loves you, he shouldn't make you wait. Do what you want with your life, make the big plans in your life without factoring him in. Just live YOUR life. It is YOUR life, him Not included. You have no obligation to wait around for him. If he wants you in his life, then he will make his plans around you.
Have an opinion?


text: We decided this time around we will use our time share holidays with dalmia resorts in Tirupati . We had to visit the temple for our venduthal . We travelled to the resort with our parents . As we had earlier travelled to this resort companies other property in Mussorie we knew what to expect from this resort and unfortunately... More


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Originally written on 5thRound.com | Last updated 10/31/12
Move over, Jon Jones and Anderson Silva. Reigning UFC heavyweight champ Junior dos Santos (Pictured) might be the next mixed martial artist to don the world famous swoosh inside the Octagon. On this week's edition of FUEL TV's UFC Tonight, it was reported dos Santos is close to signing with Nike. Helping add fuel to the fire was the fact dos Santos was sporting a Nike T-shirt during his interview on Tuesday's show. If the report holds true, 2Santos will join Jones and Silva as the only UFC fighters to endorse the most popular athletic label on the planet. "Bones" and "The Spider" reportedly have global agreements. No word if dos Santos will receive that kind of exposure or would simply represent Nike in his homeland of Brazil. Dos Santos has been sponsored by Pretorian in his last two fights – in November's championship clinching knockout victory over Cain Velasquez and in his only title defense when he put Frank Mir down at UFC 146. The likeable slugger will defend his hardware against Velasquez on December 29th at UFC 155 in Las Vegas.
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Need to know: CV cliches Why you can`t find a jobHF interviews
Facebook is spending its wealth.
The social networking giant has reached an agreement to buy Instagram for $1 billion in cash and stock. That means even before it launches its massive IPO, Facebook is using its market wealth to grow. (A post that at 11 minutes old had over 34,000 likes on it. )
Here's what Mark Zuckerberg has posted on his own "Timeline" on Facebook.
News roundup from Australian and offshore newspapers, newswires and websites on Monday, March 12, 2012. Deal Journal Australia has not verified the stories from other media organizations and does not vouch for their accuracy.
Glencore in £3.5bn bid for Canadian grain giant Viterra
Glencore has made what is believed to be a £3.5bn approach for Canadian grain giant Viterra in an attempt to bolster its agricultural business in North America. [The Telegraph]
Financing to Value Instagram at $500 Million
Instagram is poised to raise a new round of financing that will value the popular photo-sharing application at as much as $500 million, people familiar with the matter said, despite some skepticism that the fast-growing company will find a way to make money. [WSJ]
Deal Journal Australia is an up-to-the-minute take on the deals and deal makers that shape the Australian landscape, including mergers and acquisitions, capital raisings, private equity and debt markets. In short, wherever money changes hands. Deal Journal Australia is updated throughout each trading day with exclusive commentary, analysis, data, news flashes and profiles. The Wall Street Journal's Gillian Tan is the lead writer, with contributions from other Journal and Dow Jones reporters and editors. Send news items, comments and questions to email@example.com.
Content engaging our readers now, with additional prominence accorded if the story is rapidly gaining attention. Our WSJ algorithm comprises 30% page views, 20% Facebook, 20% Twitter, 20% email shares and 10% comments.
FINS has become a part of DHI in order to improve your job search experience. eFinancialCareers will provide you with the best possible service, the best job opportunities and editorial content and the best talent. With our long track record of helping finance professionals develop their career, from graduating to landing top positions in the industry, we will continue to deliver.
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