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This invention relates to compositions for and methods of treating, preventing or ameliorating cancer and other proliferative diseases as well as methods of inducing wound healing, treating cutaneous ulcers, treating gastrointestinal disorders, treating blood disorders such as anemias, immunomodulation, enhancing recombinant gene expression, treating insulin-dependent patients, treating cystic fibrosis patients, inhibiting telomerase activity, treating virus-associated tumors, especially EBV-associated tumors, augmenting expression of a tumor suppressor gene and inducing tolerance to an antigen. The methods of the invention use tricarboxylic acid substituted oxyalkyl esters.
1. A compound represented by the formula: ##STR19## wherein R is n-propyl, isopropyl, 1-methylpropyl, n-butyl, isobutyl, 1-methylbutyl, 2-methylbutyl, n-amyl, isoamyl, 1-, 2-, or 3-methylamyl, 2-ethylbutyl or benzyl, any of which can be optionally substituted with halo, trifluoromethyl, amino, alkoxy or aryl; or R is C3 to C5 linear alkenyl, optionally substituted with halo, trifluoromethyl, amino, alkoxy or aryl;
R1 and R2 are independently H, lower alkyl or lower alkenyl;
A is arylene or C1 to C8 alkylene, alkenylene or alkynylene, wherein alkylene, alkenylene and alkynylene are optionally substituted with one or more of hydroxy, halo, lower alkyl, alkoxy, carbonyl, thiol, lower alkylthio or aryl;
R3 is H, lower alkyl, aryl, or --CR1 R2 --O--C(O)--R; or
a pharmaceutically-acceptable salt thereof.
2. The compound of claim 1, wherein
R is n-propyl, 3-(phenyl)propyl, 2-propenyl or benzyl;
A is arylene or C1 to C8 alkylene, alkenylene or alkynylene, wherein alkylene, alkenylene and alkynylene are optionally substituted with one or more of hydroxy, halo, lower alkyl, alkoxy, carbonyl, thiol, lower alkylthio or aryl;
R3 is H, lower alkyl, or --CR1 R2 --O--C(O)--R; or
a pharmaceutically-acceptable salt thereof.
3. The compound of claim 1, wherein said compound is mono-(butyroyloxymethyl)glutarate, bis-(butyroyloxymethyl)glutarate, mono-(1-butyroyloxyethyl)glutarate, or bis-(1-butyroyloxyethyl)glutarate.
4. A pharmaceutical composition comprising a therapeutically-effective amount of a compound of any one of claims 1 to 3 and a pharmaceutically-effective carrier or diluent.
5. A pharmaceutical composition comprising a therapeutically-effective amount of a compound of any one of claims 1 to 3 with a therapeutically-effective amount of a pharmaceutical agent, wherein said agent is selected from the group consisting of a cytokine, an interleukin, an anti-cancer agent or anti-neoplastic agent, a chemotherapeutic agent, an antibody, a conjugated antibody, an immune stimulant, an antibiotic, a hormone antagonist or a growth stimulant.
6. The composition of claim 5 wherein said pharmaceutical agent comprises a cytotoxic agent.
7. The composition of claim 5 wherein said antibiotic is an antiviral nucleoside antibiotic selected from the group consisting of ganciclovir, acyclovir, and famciclovir.
8. The composition of claim 5 wherein said antibiotic is ganciclovir.
9. The composition of claim 5 wherein said chemotherapeutic agent is selected from the group consisting of alkylating agents, purine and pyrimidine analogs, vinca and vinca-like alkaloids, etopsides and etopside-like drugs, corticosteroids, nitrosoureas, antimetabolites, platinum based cytotoxic drugs, hormonal antagonists, anti-androgens and antiestrogens.
10. The composition of claim 5, wherein said cytokine is an interferon.
11. The composition of claim 5, wherein said immune stimulant is Corynebacterium parvum or a sarcolectin.
12. The composition of claim 5, wherein the chemotherapeutic agent is selected from the group consisting of tamoxifen, doxorubicin, 1-asparaginase, decarbazine, amacrine, procarbazine, hexamethylmelamine, mitosantrone and gemcitabine.
This invention relates to compositions for and methods of treating, preventing or ameliorating cancer and other proliferative diseases as well as methods of inducing wound healing, treating cutaneous ulcers, treating gastrointestinal disorders, treating blood disorders such as anemias, immunomodulation, enhancing recombinant gene expression, treating insulin-dependent patients, treating cystic fibrosis patients, inhibiting telomerase activity, treating virus-associated tumors, especially EBV-associated tumors, augmenting expression of tumor suppressor genes and inducing tolerance to antigens. The methods of the invention use tricarboxylic acid substituted oxyalkyl esters.
Butyric acid (BA) is a natural product. It is supplied to mammals from two main sources: 1) the diet, mainly from dairy fat, and 2) from the bacterial fermentation of unabsorbed carbohydrates in the colon, where it reaches mM concentrations (Cummings, Gut 22:763-779, 1982; Leder et al., Cell 5:319-322, 1975).
BA has been known for nearly the last three decades to be a potent differentiating and antiproliferative agent in a wide spectra of neoplastic cells in vitro (Prasad, Life Sci. 27:1351-1358, 1980). In cancer cells, BA has been reported to induce cellular and biochemical changes, e.g., in cell morphology, enzyme activity, receptor expression and cell-surface antigens (Nordenberg et al., Exp. Cell Res. 162:77-85, 1986; Nordenberg et al., Br. J. Cancer 56:493-497, 1987; and Fishman et al., J. Biol. Chem. 254:4342-4344, 1979).
Although BA or its sodium salt (sodium butyrate, SB) has been the subject of numerous studies, its mode of action is unclear. The most specific effect of butyric acid is inhibition of nuclear deacetylase(s), resulting in hyperacetylation of histones H3 and H4 (Riggs, et al., Nature 263:462-464, 1977). Increased histone acetylation following treatment with BA has been correlated with changes in transcriptional activity and the differentiated state of cells (Thorne et al., Eur. J. Biochem. 193:701-713, 1990). BA also exerts other nuclear actions, including modifications in the extent of phosphorylation (Boffa et al., J. Biol. Chem. 256:9612-9621, 1981) and methylation (Haan et al., Cancer Res. 46:713-716, 1986). Other cellular organelles, e.g., cytoskeleton and membrane composition and function, have been shown to be affected by BA (Bourgeade et al., J. Interferon Res. 1:323-332, 1981). Modulations in the expression of oncogenes and suppressor genes by BA were demonstrated in several cell types. Toscani et al., reported alterations in c-myc, p53 thymidine kinase, c-fos and AP2 in 3T3 fibroblasts (Oncogene Res. 3:223-238, 1988). A decrease in the expression of c-myc and H-ras oncogenes in B16 melanoma and in c-myc in HL-60 promyelocytic leukemia was also reported (Prasad et al., Biochem. Cell Biol. 68:1250-1255, 1992; and Rabizadeh et al., FEBS Lett. 328:225-229, 1993).
BA has been reported to induce apoptosis, i.e., programmed cell death. SB has been shown to produce apoptosis in vitro in human colon carcinoma, leukemia and retinoblastoma cell lines (Bhatia et al., Cell Growth Diff. 6:937-944, 1995; Conway et al., Oncol. Res. 7:289-297, 1993; Hague et al.; Int. J. Cancer 60:400-406, 1995). Apoptosis is the physiological mechanism for the elimination of cells in a controlled and timely manner. Organisms maintain a delicate balance between cell proliferation and cell death, which when disrupted can tip the balance between cancer, in the case of over accumulation of cells, and degenerative diseases, in the case of premature cell losses. Hence, inhibition of apoptosis can contribute to tumor growth and promote progression of neoplastic conditions.
The promising in vitro antitumor effects of BA and BA salts led to the initiation of clinical trials for the treatment of cancer patients with observed minimal or transient efficacy. [Novogrodsky et al., Cancer 51:9-14, 1983; Rephaeli et al., Intl. J. Oncol. 4:1387-1391, 1994; Miller et al., Eur. J. Cancer Clin. Oncol. 23:1283-1287, 1987].
Clinical trials have been conducted for the treatment of β-globin disorders (e.g., β-thalassemia and sickle-cell anemia) using BA salts. The BA salts elevated expression of fetal hemoglobin (HbF), normally repressed in adults, and favorably modified the disease symptoms in these patients (Stamatoyannopouos et al., Ann. Rev. Med. 43:497-521, 1992). In this regard, arginine butyrate (AB) has been used in clinical trials with moderate efficacy (Perrine et al., N. Eng. J. Med. 328:81-86, 1993; Sher et al., N. Eng. J. Med. 332:1606-1610, 1995). The reported side effects of AB included hypokalemia, headache, nausea and vomiting in β-thalassemia and sickle-cell anemia patients.
Butyric acid derivatives with antitumor activity and immunomodulatory properties have been reported in U.S. Pat. No. 5,200,553 and by Nudelman et al., 1992, J. Med. Chem. 35:687-694. The most active butyric acid prodrug reported in these references was pivaloyloxymethyl butyrate (AN-9). None of the compounds disclosed in these references included carboxylic acid-containing oxyalkyl compounds of this invention.
BA and/or its analogues have also been reported to increase the expression of transfected DNA (Carstea et al., 1993, Biophys. Biohem. Res. Comm. 192:649; Cheng et al., 1995, Am. J. Physical 268:L615-L624) and to induce tumor-restricted gene expression by adenovirus vectors (Tang et al., 1994, Cancer Gene Therapy 1:15-20). Tributyrin has been reported to enhance the expression of a reporter gene in primary and immortalized cell lines (Smith et al., 1995, Biotechniques 18:852-835).
However, BA and its salts are normally metabolized rapidly and have very short half-lives in vivo, thus the achievement and maintenance of effective plasma concentrations are problems associated with BA and BA salts, particularly for in vivo uses. BA and BA salts have required large doses to achieve even minimal therapeutic effects. Because of the high dosage, fluid overload and mild alkalosis may occur. Patients receiving BA emanate an unpleasant odor that is socially unacceptable.
While BA salts have been shown to increase HbF expression, and appear to hold therapeutic promise with low toxicity in cancer patients, they nevertheless have shown low potency in in vitro assays and clinical trials. There also remains a need to identify compounds as effective or more effective than BA or BA salts as differentiating or anti-proliferating agents for the treatment of cancers. Such compounds need to have higher potency than BA without the problems associated with BA (such as bad odor). Consequently, there remains a need for therapeutic compounds that either deliver BA to cells in a longer acting form or which have similar activity as BA but a longer duration of effectiveness in vivo.
The compounds of this invention address these needs and are more potent than BA or BA salts for treating cancers and other proliferative diseases, for treating gastrointestinal disorders, for wound healing and for treating blood disorders such as thalassemia, sickle cell anemia and other anemias, for modulating an immune response, for enhancing recombinant gene expression, for treating insulin-dependent patients, for treating cystic fibrosis patients, for inhibiting telomerase activity, for detecting cancerous or malignant cells, for treating virus-associated tumors, especially EBV-associated tumors, for augmenting expression of a tumor suppressor gene and for inducing tolerance to an antigen. For example, one of the advantages of the compounds of the invention is increased water solubility of the free carboxylic acids compounds of the invention and their salts, and easier administration, especially for intravenous administration.
Accordingly, one embodiment of the present invention is directed to a method of treating preventing or ameliorating cancer and other proliferative disorders using compounds represented by Formula (I): ##STR1## wherein R is benzyl, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, or C3 to C10 alkyl, alkenyl, or alkynyl, any of which can be optionally substituted with halo, trifluoromethyl, amino, hydroxy, alkoxy, carbonyl, aryl or heteroaryl;
R1 and R2 are independently H, lower alkyl or lower alkenyl;
A is aryl, heteroaryl, or C1 to C8 alkyl, alkenyl, or alkynyl, wherein alkyl, alkenyl and alkynyl are optionally substituted with one or more of hydroxy, halo, lower alkyl, alkoxy, carbonyl, thiol, lower alkylthio, aryl or heteroaryl;
R3 is H, alkyl, alkenyl, aryl, aralkyl, heteroaryl, heteroaralkyl, or --CR1 R2 --O--C(O)--R; and
pharmaceutically-acceptable salts thereof.
The above compounds can be used in all the methods of the invention. In a preferred embodiment, the compounds used in this and the other methods of the invention are those of Formula I, wherein
R is n-propyl; benzyl; 3-(phenyl)propyl; 2-chloroethyl; 2-propenyl; 2-(3-pyridyl)ethyl, 2-, 3- or 4-pyridylmethyl; or 3-(2-, 3- or 4-pyridyl)propyl;
A is C2 to C6 alkyl with up to three methyl substituents;
R3 is H or lower alkyl, or --CR1 R2 --O--C(O)--R; and
pharmaceutically-acceptable salts thereof.
In a more preferred embodiment, the compounds used in this and the other methods of the invention are mono-(butyroyloxymethyl)glutarate (GOMB), bis-(butyroyloxymethyl)glutarate (bisGOMB), mono-(1-butyroyloxyethyl)glutarate, bis-(1-butyroyloxyethyl)glutarate, and most preferably, GOMB.
The methods of the present invention are particularly useful for treating, preventing or ameliorating the effects of cancer and other proliferative disorders by acting as anti-proliferative or differentiating agents in subjects afflicted with such anomalies. Such disorders include but are not limited to leukemias, such as acute promyelocytic leukemia, acute myeloid leukemia, and acute myelomonocytic leukemia; other myelodysplastic syndromes, multiple myeloma such as but not limited to breast carcinomas, cervical cancers, melanomas, colon cancers, nasopharyngeal carcinoma, non-Hodgkins lymphoma (NHL), Kaposi's sarcoma, ovarian cancers, pancreatic cancers, hepatocarcinomas, prostate cancers, squamous carcinomas, other dermatologic malignancies, teratocarcinomas, T-cell lymphomas, lung tumors, gliomas, neuroblastomas, peripheral neuroectodermal tumors, rhabdomyosarcomas, and prostate tumors and other solid tumors. It is also possible that compounds of Formula (I) have anti-proliferative effects on non-cancerous cells as well, and may be of use to treat benign tumors and other proliferative disorders such as psoriasis. Preferred is the method for treating or ameliorating leukemia, squamous cell carcinoma and neuroblastoma.
Another embodiment of the present invention is directed to methods of treating, preventing or ameliorating cancer and other proliferative disorders by administering a therapeutically-effective amount of a compound of Formula (I) to a subject suffering from such disorders together with a pharmaceutical agent (e.g., a known antiproliferative, differentiating or oncostatic agents) to thereby enhance the action of these agents. Such agents include but are not limited to, cytokines, interleukins, anti-cancer agents, chemotherapeutic agents, antibodies, conjugated antibodies, immune stimulants, antibiotics, hormone antagonists, and growth stimulants. The compounds of the invention can be administered prior to, after or concurrently with any of the agents.
Yet another embodiment of the invention is directed to a method of ameliorating the effects of a cytotoxic agent which comprises adminstering a therapeutically-effective amount of an cytotoxic agent with a compound of Formula I to a mammalian patient for a time and in an amount to induce growth arrest of rapidly-proliferating epithelial cells of the patient and thereby protect those cells from the cytotoxic effects of the agent. The cytotoxic agent can be a chemotherapeutic agent, an anticancer agent, or radiation therapy. Rapidly proliferating rapidly-proliferating epithelial cells are found in hair follicles, the gastrointestinal tract, the bladder and the bone marrow, for example. Such cells include bone marrow stem cells, hair follicle cells, or intestinal cryt cells. In accordance with the invention the cytotoxic agent and the compound of Formula I can be administered simultanously, or the cytotoxic agent can be administered prior to or after the compound of the invention. Administration (simulataneously or separately) can be done systemically or topically as determined by the indication. In addition, when the cytotoxic agent is radiation therapy, the compounds of the invention can be administered to a cancer patient pre- or post-radiation therapy to treat or ameliorate the effects of cancer.
A still further embodiment of the invention is directed to a method of inducing wound healing, treating cutaneous ulcers or treating a gastrointestinal disorder by administering a therapeutically-effective amount of a compound of Formula (I) to a subject in need of such treatment. The cutaneous ulcers which can be treated in accordance with the methods of the invention include leg and decubitus ulcers, stasis ulcers, diabetic ulcers and atherosclerotic ulcers. With respect to wound healing, the compounds are useful in treating abrasions, incisions, burns, and other wounds. Gastrointestinal disorders treatable by the methods of the invention include colitis, inflammatory bowel disease, Crohn's disease and ulcerative colitis.
The invention is further directed to a method of treating blood disorders by administering a therapeutically-effective amount of a compound of Formula (I) to a patient. The blood disorders treatable in accordance with the invention include, but are not limited to, thalassemias, sickle cell anemias, infectious anemias, aplastic anemias, hypoplastic and hypoproliferative anemias, sideroblastic anemias, myelophthisic anemias, antibody-mediated anemias, anemias due to chronic diseases and enzyme-deficiencies, and anemias due to blood loss, radiation therapy and chemotherapy. In this regard, these methods can include increasing hemoglobin content in blood by administering a therapeutically-effective amount of a compound of Formula (I) to a subject.
Another embodiment of the invention is directed to a method of modulating an immune response in a host by administering an amount of a compound of Formula I effective to modulate said immune response. Modulation of the immune response includes enhancing cytokine secretion, inhibiting or delaying apoptosis in polymorphonuclear cells, enhancing polymorphonuclear cell function by augmenting hematopoietic growth factor secretion, inducing expression of cell surface antigens in tumor cells, and enhancing progenitor cell recovery after bone marrow transplantation.
A further embodiment of the invention relates to a method of enhancing recombinant gene expression by treating a recombinant host cell containing an expression system for a mammalian gene product of interest with an expression-enhancing amount of a compound of Formula I, wherein said gene product is encoded by a butyric acid-responsive gene. The host cells can be mammalian cells, insect cells, yeast cells or bacterial cells and the correspondingly known expression systems for each of these host cells. The gene product can be any protein or peptide of interest expression of which can be regulated or altered by butyric acid or a butyric acid salt. A butyric acid-responsive gene is a gene that has a promoter, enhancer element or other regulon that modulates expression of the gene under its control in response to butyric acid or a salt of butyric acid. For example, gene products contemplated for regulation in accordance with the invention include but are not limited to tumor suppressor genes (such as p53) and the γ-globin chain of fetal hemoglobin.
Yet a further embodiment of the invention is directed to a method of treating, preventing or ameliorating symptoms in insulin-dependent patients by administering an amount of a compound of Formula I effective to enhance insulin expression.
Yet another embodiment of the invention relates to a method of treating, preventing or ameliorating symptoms in cystic fibrosis patients by administering an amount of a compound of Formula I effective to enhance chloride channel expression.
Still another method of the invention is directed to a method of inhibiting telomerase activity in cancer cells by administering a telomerase-inhibiting amount of a compound of Formula I to the cells, wherein the amount is effective to decrease the telomerase activity of the cells and thereby inhibit the malignant progression of the cells. This method can be applied to in vivo or in vitro cells.
Another embodiment of this invention is directed to a method of treating, preventing or ameliorating virus-associated tumors by pre-, post or co-administering a therapeutically-effective amount of a compound of Formula I with a therapeutically-effective amount of an antiviral agent. Antiviral agents contemplated for use in the invention include ganciclovir, acyclovir and famciclovir, and preferably ganciclovir. The virus-associated tumors which can be treated, prevented or ameliorated in accordance with the invention include, but are not limited to, EBV-associated malignancy, Kaposi's sarcoma, AIDS-related lymphoma, hepatitis B-associated malignancy or hepatitis C associated malignancy. EBV-associated malignancies include nasopharyngeal carcinoma and non-Hodgkins' lymphoma and are preferred embodiments of the invention.
Further still the invention is directed to a method of augmenting gene expression, especially of a tumor suppressor gene, a butyric acid-responsive gene or a fetal hemoglobin gene, by treating a host or host cells with an expression-enhancing amount of a compound of Formula I. Preferably the host is a cancer patient. This method of the invention thus includes augmenting tumor suppressor gene expression in conjunction with ex vivo or in vivo gene therapy, i.e., the compound of the invention can be co-administered to the host during administration of gene therapy vectors or administration of the ex vivo transfected cells. Similarly, the compounds of the invention can be used to treat cells during the transfection step of ex vivo gene therapy. The hosts of the method therefore include cancer patients or other patients under going gene therapy. The host cells of the invention include hematopoietic cells such as stem cells and progenitor cells, e.g., or any other cell type used in ex vivo gene therapy.
Yet another embodiment of the invention is directed to a method of inducing tolerance to an antigen which comprises administering a therapeutically-effective amount of compound of Formula I. Preferably the antigen is a self-antigen.
Another aspect of the invention is directed to compounds represented by Formula (II): ##STR2## wherein R is n-propyl, isopropyl, 1-methylpropyl, n-butyl, isobutyl, 1-methylbutyl, 2-methylbutyl, n-amyl, isoamyl, 1-, 2- or 3-methylamyl, 2-ethylbutyl, benzyl, 2-pyridylmethyl, 3-pyridylmethyl, or 4-pyridylmethyl, any of which can be optionally substituted with halo, trifluoromethyl, amino, alkoxy, aryl, or heteroaryl; or R is C3 to C5 linear alkenyl, optionally substituted with halo, trifluoromethyl, amino, alkoxy, aryl, or heteroaryl;
R1 and R2 are independently H, lower alkyl or lower alkenyl;
A is aryl, heteroaryl, or C1 to C8 alkyl, alkenyl, or alkynyl, wherein alkyl, alkenyl and alkynyl are optionally substituted with one or more of hydroxy, halo, lower alkyl, alkoxy, carbonyl, thiol, lower alkylthio, aryl or heteroaryl;
R3 is H, lower alkyl, aryl, or --CR1 R2 --O--C(O)--R; and
pharmaceutically-acceptable salts thereof.
In a preferred embodiment, the compounds of the invention are those of Formula II,
R is n-propyl, 3-(phenyl)propyl, 2-propenyl, benzyl, 2-pyridylmethyl, 3-pyridylmethyl, or 4-pyridylmethyl or 3-(2-, 3- or 4-pyridyl)propyl;
A is C1 to C8 alkyl, alkenyl, or alkynyl, optionally substituted with halo, alkoxy, aryl, substituted aryl, heteroaryl, or substituted heteroaryl;
R3 is H, lower alkyl, or --CR1 R2 --O--C(O)--R; and
pharmaceutically-acceptable salts thereof.
In a more preferred embodiment, the compounds of the invention are GOMB, bisGOMB, mono-(1-butyroyloxyethyl)glutarate and bis-(1-butyroyloxyethyl)glutarate, and most preferably, GOMB.
Another embodiment of the present invention is drawn to pharmaceutical compositions comprising a therapeutically-effective amount of a compound of Formula (II) and a pharmaceutically-effective carrier or diluent.
A further embodiment of the present invention is directed to pharmaceutical compositions comprising a therapeutically-effective amount of a compound of Formula (II) with a pharmaceutical agent and a pharmaceutically-effective carrier or diluent. The pharmaceutical agents of the invention include but are not limited to cytokines, interleukins, anti-cancer agents, chemotherapeutic agents, antibodies, conjugated antibodies, immune stimulants, antibiotics, hormone antagonists or growth stimulants.
FIG. 1 is a graphic illustration showing the in vitro inhibition of cellular growth (clonogenicity) by GOMB and AB on established human neuroblastoma cell lines as a function of concentration (in μM).
FIG. 2 is a graphic illustration showing the average in vitro inhibition of growth by GOMB and AB on 53 primary human tumor cells as a function of concentration (in μM).
FIG. 3 is a graphic illustration showing growth inhibition measured by reduction of [3 H]-thymidine uptake in EBV-transformed cell line as a function of concentration of ganciclovir (GC)(panel A), GC and AB (panel B) and GC and GOMB (panel C). In panels B and C, the GC concentration is indicated in the inset.
FIG. 4 is a bar graph showing the percentage of NBT-positive human leukemia cells for cells treated with AB, AN-9 and GOMB.
FIG. 5 is a bar graph comparing the IC50 and IC90 values of GOMB and AN-9 on the indicated cell lines treated in the SRB assay.
The compounds herein described may have asymmetric centers. All chiral, diastereomeric, and racemic forms are included in the present invention. Many geometric isomers of olefins and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present invention.
By "stable compound" or "stable structure" is meant herein a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.
As used herein, "alkyl" means both branched- and straight-chain, saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. As used herein "lower alkyl" means an alkyl group having 1 to 5 carbon atoms. As used herein, "alkenyl" means hydrocarbon chains of either a straight or branched configuration and one or more unsaturated carbon-carbon bonds, such as ethenyl, propenyl, and the like. Lower alkenyl is an alkenyl group having 2 to 6 carbon atoms. As used herein, "alkynyl" means hydrocarbon chains of either a straight or branched configuration and one or more carbon-carbon triple bonds, such as ethynyl, propynyl and the like.
When alkyl, alkenyl, and alkynyl, or a variation thereof such as lower alkyl, are used to define the A moiety of Formulas I and II (or any other group that has two appendages) then these groups have the meanings as defined herein except that each term refers to a hydrocarbon chain having the specified number of carbon atoms. Similarly, when aryl and heteroaryl are used to define the A moiety of Formulas I and II, then these groups have the meanings defined herein except that there are two attachment points on the aromatic ring. When A is an alkyl chain, one preferred A moiety has up to three lower alkyl substituents and, even more preferably, up to three methyl groups.
As used herein, "aryl" includes "aryl" and "substituted aryl." Thus "aryl" of this invention means any stable 6- to 14-membered monocyclic, bicyclic or tricyclic ring, containing at least one aromatic carbon ring, for example, phenyl, naphthyl, indanyl, tetrahydronaphthyl (tetralin) and the like, optionally substituted with halo, alkyl, alkoxy, hydroxy, carboxy, cyano, nitro, amino or acylamino.
As used herein, the term "heteroaryl" includes "heteroaryl" and "substituted heteroaryl." Thus "heteroaryl" of this invention means a stable 5- to 10-membered monocyclic or bicyclic heterocyclic ring which is aromatic, and which consists of carbon atoms and from 1 to 3 heteroatoms selected from the group consisting of N, O and S and wherein the nitrogen may optionally be quaternized, and including any bicyclic group in which any of the above-defined heteroaryl rings is fused to a benzene ring. The heteroaryl ring may be attached to its pendant group at any heteroatom or carbon atom which results in a stable structure. The heteroaryl rings described herein may be optionally substituted on carbon, on a nitrogen atom or other heteroatom if the resulting compound is stable and all the valencies of the atoms have been satisfied. The substituents of the substituted heteroaryl groups are as for the substituted aryl groups (see above). Examples of heteroaryl groups include, but are not limited to, pyridyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl, benzofuranyl, benzothienyl, indolyl, indolenyl, quinolinyl, isoquinolinyl and benzimidazolyl.
As used herein and in the claim, "aralkyl", and "heteroaralkyl" refer to an aryl or heteroaryl group, respectively, as described above attached to an alkyl group as described above. Examples of heteroaralkyl groups include but are not limited to 2-, 3-, or 4-pyridylmethyl and 3-(2-, 3- or 4- pyridyl)propyl and the like.
The term "substituted", as used herein, means that one or more hydrogens on the designated atom are replaced with a selection from the indicated substituents, provided that the designated atom's normal valency is not exceeded, and that the substitution results in a stable compound.
The substituents of the invention include, as indicated, halo, hydroxy, alkyl, alkoxy, amino, acylamino, carboxy, carbonyl, cyano, nitro, and trifluoromethyl groups. These groups can be substituents for alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and saturated heterocyclic groups as indicated in accordance with the invention. A halo group is a halogen, and includes fluoro, chloro, bromo and iodo groups. The alkyl moiety of alkoxy, acyl, aralkyl, heteroaralkyl and the like is lower alkyl unless otherwise specified.
As used herein, "therapeutically-effective amount" refers to that amount necessary to administer to a host to achieve an anti-tumor effect; to induce differentiation and/or inhibition of proliferation of malignant cancer cells, benign tumor cells or other proliferative cells; to aid in the chemoprevention of cancer; to promote wound healing; to treat a gastrointestinal disorder; to treat a blood disorder or increase the hemoglobin content of blood; to modulate an immune response; to enhance gene expression; to augment expression of tumor suppressor genes; to enhance insulin expression; to enhance chloride channel expression or to induce tolerance to an antigen. Methods of determining therapeutically-effective amounts are well known.
As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds that are modified by making acid or base salts. Examples 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, and the like.
Pharmaceutically-acceptable salts of the compounds of the invention can be 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. The salts of the invention can also be prepared by ion exchange, for example. 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 in its entirety.
The "pharmaceutical agents" for use in the methods of the invention related to the coadministration of compounds of Formula I and Formula II, include but are not limited to anticancer agents as well as differentiating agents. For example, the pharmaceutical agent can be a cytokine, an interleukin, an anti-cancer agent or anti-neoplastic agent, a chemotherapeutic agent, an antibody, a conjugated antibody, an immune stimulant, an antibiotic, a hormone antagonist or a growth stimulant. The pharmaceutical agent can also be a cytotoxic agent. Cytotoxic agents include antiviral nucleoside antibiotic such as ganciclovir, acyclovir, and famciclovir.
As used herein, the "chemotherapeutic agents" include but are not limited to alkylating agents, purine and pyrimidine analogs, vinca and vinca-like alkaloids, etoposide and etoposide-like drugs, corticosteroids, nitrosoureas, antimetabolites, platinum-based cytotoxic drugs, hormonal antagonists, anti-androgens and antiestrogens.
The "cytokines" for use herein include but are not limited to interferon, preferably α, β or γ interferon, as well as IL-2, IL-3, G-CSF, GM-CSF and EPO.
As used herein, an "immune stimulant" is a substance such as C. parvum or sarcolectin which stimulates a humoral or cellular component of the immune system.
The chemotherapeutic agents of the invention include but are not limited to tamoxifen, doxorubicin, 1-asparaginase, decarbazine, amacrine, procarbazine, hexamethylmelamine, mitosantrone and gemcitabine.
The compounds of the present invention can generally be prepared by any method known in the art. For example, the compounds of the invention can be made by reacting the acid form of the RCOOH with a reagent of the formula ##STR3## or by similar reactions between any of the appropriate acids and the appropriate alkyl halides, in the presence of a base, where Y is a leaving group such as halogen, methane sulfonate or p-toulenesulfonate and R, R1, R2 and R3 are as defined herein. The above reagents are readily prepared according to literature procedures, see for example, Nudelman et al., J. Med. Chem. 35:687-694, 1992, and Japanese patent 07033709 (1995). The base can be a trialkylamine, pyridine, an alkali metal carbonate or other suitable base. The reaction can be carried out in the presence or absence of an inert solvent. Suitable solvents include, for example, acetone, benzene, toluene, tetrahydrofuran, ethyl acetate, acetonitrile, dimethylformamide, dimethyl sulfoxide, chloroform, dioxan or 1,2-dichloroethane.
The procedures outlined above can be improved by one skilled in the art by, for instance, changing the temperature, duration, stoichiometry or other parameters of the reactions. Any such changes are intended to fall within the scope of this invention.
The activities of the compounds of the invention can be measured using generally-accepted techniques known to those skilled in the art consistent with the activity of interest. For example, the activity of compounds useful as differentiating agents can be measured using standard methodology of the nitro-blue tetrazolium reduction assay (e.g., Rabizadeh et al., FEBS Lett. 328:225-229, 1993; Chomienne et al., Leuk. Res. 10:631, 1986; and Breitman et al. in Methods for Serum-free Culture of Neuronal and Lymphoid Cells, Alan R. Liss, New York, p. 215-236, 1984 which are hereby incorporated by reference in their entirety) and as described below. This in vitro assay has been deemed to be predictive and in fact correlative with in vivo efficacy (Castaigne et al., Blood 76:1704-1709, 1990).
Another assay which is predictive of differentiating activity is the morphological examination for the presence of Auer rods and/or specific differentiation cell surface antigens in cells collected from treatment groups, as described in Chomienne et al., (Blood 76:1710-1717, 1990 which is hereby incorporated by reference in its entirety) and as described below.
The compounds of the present invention also have anti-proliferative and anti-tumor activity. The anti-proliferation activity of compounds of the present invention can be determined by methods generally known to those skilled in the art. Generally-accepted assays for measuring viability and anti-proliferative activity are the trypan blue exclusion test and incorporation of tritiated thymidine, also as described by Chomienne, et al., above, which is incorporated herein by reference. Other assays which predict and correlate antitumor activity and in vivo efficacy are the human tumor colony forming assay described in Shoemaker et al., Can. Res. 45:2145-2153, 1985, and inhibition of telomerase activity as described by Hiyayama et al., J. Natl. Cancer Inst. 87:895-908, 1995, which are both incorporated herein by reference in their entirety. These assays are described in further detail below.
Cell Cultures
Human promyelocytic leukemia cells (HL-60), human pancreatic carcinoma cells (PaCa-2) and human breast adenocarcinoma cells, pleural effusion cells (MCF-7) can be cultured as follows. Cells are grown in RPMI media with 10% FCS, supplemented with 2 mM glutamine and incubated at 37� C. in a humidified 5% CO2 incubator. Alternatively, cells can be grown in any other appropriate growth medium and conditions which supports the growth of the cell line under investigation. Viability can be determined by trypan blue exclusion. Cells are exposed to a test compound, cultures are harvested at various time points following treatment and stained with trypan blue.
Cellular Staining to Detect Differentiation
Lipid staining and/or immunochemical staining of casein can be used as a marker for cellular differentiation of breast cancer cells (Bacus et al., Md. Carcin. 3:350-362, 1990). Casein detection can be done by histochemical staining of breast cancer cells using a human antibody to human casein as described by Cheung et al., J. Clin. Invest. 75:1722-1728, which is incorporated by reference in its entirety.
Nitro-Blue Tetrazolium (NBT) Assay
Cell differentiation of myeloid leukemia cells can be evaluated, for example, by NBT reduction activity as follows. Cell cultures are grown in the presence of a test compound for the desired time period. The culture medium is then brought to 0.1% NBT and the cells are 21 stimulated with 400 mM of 12-O-tetradecanoyl-phorbol-13-acetate (TPA). After incubation for 30 min at 37� C., the cells are examined microscopically by scoring at least 200 cells. The capacity for cells to reduce NBT is assessed as the percentage of cells containing intracellular reduced black formazan deposits and corrected for viability.
Cell Surface Antigen Immunophenotyping
Cell surface antigen immunotyping can be conducted using dual-color fluorescence of cells gated according to size. The expression of a panel of antigens from early myeloid (CD33) to late myeloid can be determined as described in Warrell, Jr. et al., New Engl. J. Med. 324:1385-1392, 1992, which is incorporated by reference herein in its entirety.
Apoptosis Evaluation
Apoptosis can be evaluated by DNA fragmentation, visible changes in nuclear structure or immunocytochemical analysis of Bcl-2 expression.
DNA fragmentation can be monitored by the appearance of a DNA ladder on an agarose gel. For example, cellular DNA is isolated and analyzed by the method of Martin et al., J. Immunol., 145:1859-1867, 1990 which is incorporated by reference herein in its entirety.
Changes in nuclear structure can be assessed, for example, by acridine orange staining method of Hare et al., J. Hist. Cyt., 34:215-220, 1986 which is incorporated by reference herein in its entirety.
Immunological detection of Bcl-2 can be performed on untreated cells and cells treated with the test compound. HL-60 cells are preferred but other cell lines capable of expressing Bcl-2 can be used. Cytospins are prepared and the cells are fixed with ethanol. Fixed cells are reacted overnight at 4� C. with the primary monoclonal antibody, anti-Bcl-2 at a dilution of 1:50. Staining is completed to visualize antibody binding, for example, using Strep A-B Universal Kit (Sigma) in accordance with the manufacturer's instructions. Identically-treated cells which received no primary antibody can serve as a non-specific binding control. Commercial kits are also available and can be used for detecting apoptosis, for example, Oncor's Apop Tag�.
Modulation of Gene Expression
The levels of expression from oncogene and tumor suppressor genes can be evaluated by routine methods known in the art such as Northern blotting of RNA, immunoblotting of protein and PCR amplification.
Mouse Cancer Model
Compounds can be examined for their ability to increase the life span of animals bearing B16 melanomas, Lewis lung carcinomas and myelomonocytic leukemias as described in Nudelman et al., J. Med. Chem. 35:687-694, 1992, or Rephaeli et al., Int. J. Cancer 49:66-72, 1991, which are incorporated by reference herein in their entireties.
For example, the efficacy of compounds of the present invention in a leukemia model can be tested as follows: Balb/c mice are injected with WEHI cells and a test compound or control solution is administered the following day. The life span of the treated animals is compared to that of untreated animals.
The efficacy of compounds of the present invention on primary tumors can also be tested with subcutaneously implanted lung carcinoma or B16 melanoma by measuring the mass of the tumor at the site of implantation every two weeks in control and treated animals.
The efficacy of compounds in xenografts can be determined by implanting the human tumor cells subcutaneously into athymic mice. Human tumor cell lines which can be used include, but are not limited to, prostate carcinoma (human Pc-3 cells), pancreatic carcinoma (human Mia PaCa cells), colon adenocarcinoma (human HCT-15 cells) and mammary adenocarcinoma (human MX-1 cells). Treatment with control solution or a test compound of the invention begins, for example, when tumors are approximately 100 mg. Anti-tumor activity is assessed by measuring the delay in tumor growth, and/or tumor shrinking and/or increased survival of the treated animals relative to control animals.
Telomerase Activity
High levels of telomerase activity is associated with the high proliferation rate found in cancer cells. Compounds which inhibit telomerase activity results in inhibition of cancer cell growth and de-differentiation. Commercially available telomerase assays can thus be used to assess the anticancer activities of compounds on cancer cell lines.
The chemoprevention activity of the compounds of the invention can be determined in the two-stage mouse carcinogenesis model of Nishimo et al. (supra).
Assay of Compounds
Compounds of the invention, their salts or metabolites, can be measured in a biological sample by any method known to those skilled in the art of pharmacology, clinical chemistry or the like. Such methods for measuring these compounds are standard methods and include, but are not limited to high performance liquid chromatography (HPLC), gas chromatography (GC), gas chromatography mass spectroscopy (GC-MS), radioimmunoassay (RIA), and others.
Dosage and Formulation
The compounds of the present invention can be administered to a mammalian patient to treat cancer or in any other method of the invention which involves treating a patient by any means that produces contact of the active agent with the agent's site of action in the body of the subject. Mammalian patients include humans and domestic animals. The compounds of the invention 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. The compounds can be administered alone, but are generally administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice. The pharmaceutical compositions of the invention may be adapted for oral, parenteral, transdermal, transmucosal, rectal or intranasal administration, and may be in unit dosage form, as is well known to those skilled in the pharmaceutical art. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, or intrasternal injection or infusion techniques.
The appropriate dosage administered in any given case will, of course, vary depending upon known factors, such as the pharmacodynamic characteristics of the particular agent and its mode and route of administration; the age, general health, metabolism, weight of the recipient and other factors which influence response to the compound; the nature and extent of the symptoms; the kind of concurrent treatment; the frequency of treatment; and the effect desired. A daily dosage of active ingredient can be expected to be about 10 to 10,000 milligrams per meter2 of body mass (mg/m2), with the preferred dose being 50-5,000 mg/M2 body mass.
Dosage forms (compositions suitable for administration) contain from about 1 mg to about 1 g 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.
The active ingredient can be administered orally in solid or semi-solid dosage forms, such as for example hard or soft-gelatin capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, disperse powders or granules, emulsions, and aqueous or oily suspensions. It can also be administered parenterally, in sterile liquid dosage forms. Other dosage forms include transdermal administration via a patch mechanism or ointment.
Compositions intended for oral use may be prepared according to any methods known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents including sweetening agents, flavoring agents, coloring agents, and preserving agents in order to provide a pharmaceutically elegant and palatable preparation.
Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. Such excipients may include, for example, inert diluents, such as calcium phosphate, calcium carbonate, sodium carbonate, sodium phosphate, or lactose; granulating disintegrating agents, for example, maize starch or alginic acid; binding agents, such as starch, gelatin, or acacia; and lubricating agents, for example, magnesium stearate, stearic acids or talc. Compressed tablets may be uncoated or may be sugar coated or film coated by known techniques to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration and adsorption in the gastrointestinal tract.
Hard gelatin capsules or liquid filled soft gelatin capsules contain the active ingredient and inert powdered or liquid carriers, such as, but not limited to calcium carbonate, calcium phosphate, kaolin, lactose, lecithin starch, cellulose derivatives, magnesium stearate, stearic acid, arachis oil, liquid paraffin, olive oil, pharmaceutically-accepted synthetic oils and other diluents suitable for the manufacture of capsules. Both tablets and capsules can be manufactured as sustained release-products to provide for continuous release of medication over a period of hours.
Aqueous suspensions contain the active compound in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, e.g., sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and gum acacia; dispersing or wetting agents, such as a naturally occurring phosphatide, e.g., lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or a condensation products of ethylene oxide with long chain aliphatic alcohols, e.g., heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol, e.g., polyoxyethylene sorbitol monooleate, or a condensation product of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, e.g., polyoxyethylene sorbitan monooleate. The aqueous suspensions can also contain one or more preservatives, for example ethyl, n-propyl, or p-hydroxy benzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose, saccharin, or sodium or calcium cyclamate.
Dispersable powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example, sweetening, flavoring, and coloring agents, can also be present.
Syrups and elixirs can be formulated with sweetening agents, such as glycerol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents.
The pharmaceutical compositions can be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3-butane diol.
In general, water, a suitable oil, saline, aqueous dextrose (glucose), polysorbate and related sugar solutions, emulsions, such as Intralipid� (Cutter Laboratories, Inc., Berkley Calif.) and glycols such as propylene glycol or polyethylene glycols are suitable carriers for parenteral solutions. Antioxidizing agents, such as but not limited to sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or combined, are suitable stabilizing agents. Also used can be citric acid and its salts and sodium EDTA. In addition, parenteral solutions can contain preservatives, such as but not limited to benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol.
The pharmaceutical compositions of the present invention also include compositions for delivery across cutaneous or mucosal epithelia including transdermal, intranasal, sublingual, buccal, and rectal administration. Such compositions may be part of a transdermal device, patch, topical formulation, gel, etc., with appropriate excipients. Thus, the compounds of the present invention can be compounded with a penetration-enhancing agent such as 1-n-dodecylazacyclopentan-2-one or the other penetration-enhancing agents disclosed in U.S. Pat. Nos. 3,991,203 and 4,122,170 which are hereby incorporated by reference in their entirety to describe penetration-enhancing agents which can be included in the transdermal or intranasal compositions of this invention.
Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field, which is incorporated herein by reference in its entirety.
Various modifications of the invention in addition to those shown and described herein will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.
The foregoing disclosure includes all the information deemed essential to enable those skilled in the art to practice the claimed invention. Because the cited patents or publications may provide further useful information these cited materials are hereby incorporated by reference in their entirety.
GOMB has the following structure ##STR4## and was synthesized as follows. To a stirred solution of chloromethyl butyrate (8.16g, 60 mmol) in acetone (50 mL), glutaric acid (7.9 g, 60 mmol) was added, followed by the dropwise addition of triethylamine (16 mL, 11.6 g, 120 mmol). The reaction mixture was heated at 70� C. for 2 h, and was further stirred at room temperature for 60 h, after which a large amount of precipitate was obtained. The precipitate was filtered, washed with acetone and the filtrate was partitioned between water (20 mL) and ethyl acetate (20 mL). The aqueous phase was brought to pH 8 with 1 M K2 CO3 and washed with ethyl acetate. The organic phase was extracted 3 more times with basic water (pH 8), and the washings were added to the aqueous phase. The washed organic phase (Solution A) was saved for isolation of bisGOMB. The combined aqueous phase was acidified with 2M HCl and extracted with ethyl acetate (4�20 mL). This second organic phase was collected, dried and evaporated to give the crude residue (3.44 g) which was dissolved in chloroform, washed with water and evaporated to give the pure GOMB, 3 g (22% yield).
To prepare bisGOMB, Solution A was dried and evaporated to give an oily residue(76 g) of crude bisGOMB. This residue was Kugelrohr distilled (190� C./0.5 mm Hg) to give 70 g (0.20 moles, 20% yield) of pure bisGOMB.
The compounds of Table 1 are those of Formula I having the specified groups and can be synthesized in a manner similar to that described above using the appropriate reagents.
TABLE 1__________________________________________________________________________Additional Formula I Compounds of the InventionR R1 R2 A R3__________________________________________________________________________n-C3 H7 -- CH3 H ##STR5## --C2 H5n-C3 H7 -- CH3 H ##STR6## ##STR7##C6 H5 (CH2)3 -- CH3 CH3 --CH2 --CH2 -- --H4-ClC6 H4 -- H H --CH═CH-- --CH32-pyridyl H H ##STR8## --H3-pyridyl H H ##STR9## ##STR10## ##STR11## H H --CH2 C.tbd.C--CH2 -- --H ##STR12## CH3 C2 H5 --(CH2)8 -- --CH3ClCH2 CH2 -- H H ##STR13## --C6 H5CH2 ═CHCH2 -- H CH3 --(CH2)3 -- --HCH3 --CH═CH2 -- n-C3 H7 H ##STR14## Hn-C3 H7 -- CH3 H ##STR15## ##STR16##C6 H5 CH2 -- C4 H9 H ##STR17## ##STR18##__________________________________________________________________________
Inhibition of tumor growth was tested using cell lines as follows:
The cell lines listed in Table 2 were grown to 70-80% confluence in complete media. Cells were harvested, washed in complete media, and counted. Cell viability was determined by trypan blue exclusion. The cells were placed into soft agar (0.12% in media) and plated at 5,000 viable cells per well onto an agarose underlayer (0.4%) in 24-well plates. After overnight culture, AB or GOMB was added at the indicated concentration. Control cells received media alone. As a control for cell death, cells were treated with a superlethal dose of 10 μg/ml of cisplatin. The dosage of AB or GOMB which inhibited fifty percent (or ninety percent) of cell proliferation (IC50 or IC90) was calculated using the Chou Analysis' Median Effective Dose equation.
The clonogenicity is determined as the percentage of clones in treated cultures relative to clones in media-treated controls cultures. A representative clonogenicity titration curve for AB and GOMB is shown with four neuroblastoma cell lines in FIG. 1. The IC50 and IC90 values of GOMB and AB for cancer cell lines are provided in Table 3.
The results demonstrate that GOMB is more a potent growth inhibitor than AB. The data show that GOMB and AB inhibit cell proliferation in a dose-dependent manner but that the cells are an order of magnitude more sensitive to GOMB. The ratio of IC50 AB:IC50 GOMB ranges between 6- to 460-fold with a median of 28.5 μM. Similarly the ratio of IC90 AB:IC90, GOMB ranges between 26-342-fold with a median value of 53 μM. The IC90 values are clinically important for assessing eradication of residual cancer disease.
TABLE 2______________________________________Human Tumor Cell LinesCell Lines Origin______________________________________MCF7-WT Breast CarcinomaMCF7-40F Breast CarcinomaPC3 Prostate CarcinomaLNCaP Prostate CarcinomaK562 ErytholeukemiaSK-N-SH NeuroblastomaNBAS-5 NeuroblastomaIMR-32 NeuroblastomaLA1-5S NeuroblastomaNBL-W-N NeuroblastomaSMS-KAN NeuroblastomaSK-N-MC Neuroblastoma______________________________________
TABLE 3______________________________________Inhibition of Established and Primary TumorCell Lines by AB and GOMB AB/GOMB AB GOMB IC50 IC90Cell Line IC50 .sup.(a) IC90 IC50 IC90 Ratio Ratio______________________________________MCF7-WT 1500 9473 43.5 97 34.5 97.7MCF7-40F 817 3183 65.9 85.3 12.4 37.3PC3 226 447 34.9 96.1 6.48 46.6LNCaP 780 6703 68.1 107 11.4 63K562 772 665 43.3 97 17.8 68.6SK-N-SH 998 3397 28 95 35.6 37.8NBAS-5 883 13030 16 38 23.2 343SK-N-MC 215 1314 16 35 13.4 37.5IMR-32 881 3566 23 41 38 87LA1-5S 1627 2675 39 59 42 45NBL-W-N 489 3074 25 70 19.6 44SMS-KAN 1138 2079 27 80 42 26Primary.sup.(b) 4645 2120 34 78 13.9 27.3______________________________________ .sup.(a) All concentrations are in μM. .sup.(b) Data from 53 primary tumor cell lines treated with AB and GOMB a described in Example 3.
The effect of GOMB was compared to that of AB on a variety of primary tumor cells as described in Example 2, except that,cells were seeded at 30,000 cells per well. The average IC50 value, IC90 value, AB/GOMB IC50 ratio and AB/GOMB IC90 ratio are provided in Table 3. The tested tumor cells were from 11 non-small cell lung carcinomas, 10 breast carcinomas, 10 gastric carcinomas, 10 ovarian carcinomas, and 10 CNS tumors. FIG. 2 shows the average tumor cell clonogenicity as a function of AB and GOMB for all 53 lines of tumor cells; GOMB is a more potent inhibitor of cell proliferation than is AB.
The effect of GOMB to AB was compared on EBV virus-associated tumors. EBV-positive cells from P3HR-1, a human lymphoma cell line, were incubated in growth medium with the specified concentration of GOMB, AB, ganciclovir (GC), or combination of these, for 72 hours, seeded in 96-well plates and pulsed with 1 μCi [3 H]-thymidine/well during the last 16 hours of exposure to the compounds. Cells were harvested with a cell harvester, using glass microfiber filters. The incorporation of [3 H]-thymidine was determined by retention of the acid insoluble fraction on filters and is expressed as CPM. The results of a representative experiment are shown in FIG. 3. FIG. 3A shows a titration curve using GC; FIG. 3B shows titration curves using GC and AB; and FIG. 3C shows similar titration curves for GC and GOMB; the GC concentration is indicated in the inset for panels B and C.
The results show that GOMB is 10-fold more potent than AB in inhibiting EBV-positive lymphoma cells, and importantly, that AB and GOMB interact synergistically with GC to inhibit proliferation of an EBV-associated tumor cell line.
Cancer cell differentiation was evaluated in a human leukemia cell line by nitroblue tetrazolium reduction (NBT) activity (Koeffler, Blood, 62: 709-721, 1983) or in a breast carcinoma cell line by lipid staining (Bacus et al., Mol. Carcinog. 3:350-362, 1990).
The differentiation ability of AB and GOMB with the human leukemia cell line HL-60 was compared to pivaloyoxylmethyl butyrate (AN-9). Briefly, HL-60 cells were incubated with the indicated concentration of AB, GOMB or AN-9 for three days, washed, resuspended in saline containing 0.1% NBT and stimulated with 0.4 μM phorbol ester for 30 minutes at 37� C. The cells were examined microscopically and at least 200 cells were scored. The results are provided graphically in FIG. 4 and demonstrate that GOMB is more active at inducing differentiation than either AB or AN-9.
In the case of the human breast carcinoma cell line Au565, 100%. Of the cells treated with 10 μM GOMB were positive for lipid staining whereas less than 1% of cells treated with the same concentration of AB were positive for lipid staining.
The inhibition of cell proliferation was measured in the indicated cancer cell lines using the sulforhoamine B (SRB) assay as described by Monks et al., J. Natl. Can. Inst. 83:757-766. The SRB assay is used to screen for anti-cancer drugs. A comparison of GOMB, AB and AN-9 demonstrates that GOMB exerts at least 100-fold greater activity than AB as measured by the IC50 and IC90 values (Table 4). GOMB also was 1.4- to 2.2-fold more active than AN-9 in 6 out of 8 of the IC50 and IC90 determinations. (Table 4 and FIG. 5).
TABLE 4__________________________________________________________________________Comparison of AB, GOMB and AN-9 in the SRB AssayCELL LINESCFPAC HL-60 HT-29 MCF7AGENT IC50 IC90 IC50 IC90 IC50 IC90 IC50 IC90__________________________________________________________________________AB >4.0 >4.0 1.644 3.455 >4.0 >4.0 >4.0 >4.0GOMB 0.0325 0.0410 0.0245 0.0536 0.0982 0.123 0.109 0.218AN-9 0.0559 0.0898 0.0390 0.0599 0.144 0.231 0.180 0.238GOMB/ 1.72 2.2 1.6 1.11 1.4 1.9 1.65 1.01AN9__________________________________________________________________________
Apoptosis was measured as described by Telford et al., 1991, Cell Physiol. 24:447-459, in HL-60 cells and in the human erthroblastic cell line, K-562. Cells were treated with COMB for 3 days, fixed, stained with propidium iodine and analyzed by flow cytometrly for cell cycle distribution. A distinct subpopulation of cells was observed in a region below the G0-G1 cells. This region consisted of cells with fragmented DNA. The increase in apoptotic cells in HL-60 and K-562 cells lines treated with varying concentrations of COMB is shown in Table 5.
Induction of apoptosis by COMB and AB in MCF-7 cells was evaluated by staining cells with the WAF antibody and by Western blot analysis as described by Bacus et al., 1996, Oncogene 12:2535-2547. The percentage of cells expressing WAF was 13% in the controls and increased 623% in GOMB-treated cells. The results demonstrate that GOMB induces apoptosis in a dose-dependent manner and that COMB induces apoptosis at a 10-fold lower concentration than does AB(Table 6).
TABLE 5______________________________________Induction of Apaptosis in Human Leukemia CellsGOMB % apoptotic cells % apoptotic cells(μM) HL-60 K-562______________________________________ 5.0 9.4 ND*12.5 14.9 925.0 15.8 1837.5 36.3 ND______________________________________ *ND, not determined.
TABLE 6______________________________________Apoptosis in Breast Cancer CellsConcentration % Cells Expressing WAF(μM) AB GOMB______________________________________10 ND* 3625 ND 3450 ND 79100 23 81______________________________________ *ND, not detected.
Hb Measurement: HB was measured by benzidine staining of K562 cells after 5 days exposure to GOMB or AB according to the procedure of Fibach et al. (1989) [full cite]. Quantitative measurement of HbF in K562 culture or human erythroid cultures was determined by ion-exchange high pressure liquid chromatography (HPLC) as described by Fibach et al., Blood 81:1630-1635, 1993.
K562 cells: K562 is an erthroblast cell line that develops some properties of erythroid, megakaryocyte or monocyte cells, depending on the specific stimulus, when induced by different chemicals. K562 cells were grown in RPMI with 10% FCS, supplemented with 2 mM glutamine. Cells were incubated at 37� C. in a humidified, 5% CO2 incubator.
Treatment of K562 cells with GOMB and AG showed that, on a molar basis, GOMB has a higher activity in inducing erythroid differentiation (hemoglobin accumulation) than does AB. This was evident from the higher proportion of Hb-containing cells per the total cell population (Table 7) as well as the total Hb content of the cultures (Tables 8). The extent of differentiation of the treated cultures was directly related to the drug dose. The diluents, DMF and water, had no effect on cell growth, cell viability or differentiation.
Erythroid cells: Erythroid cells were isolated from peripheral blood of healthy individuals by the method of Fibach (1993). The results showed that 0.2 mM GOMB increased HbF by 49.5% in cultured erythroid cells relative to untreated control cultures.
TABLE 7______________________________________Percentage K562 Cells Containing HbFConcentration (mM) GOMB AB______________________________________0.01 6.3 � 1.15 ND0.05 12.3 � 5.3 3 � 20.10 29 ND______________________________________
TABLE 8______________________________________HbF Synthesis in K562 CellsConcentration (mM) GOMB AB______________________________________0.01 1.54* 0.960.05 3.21 0.950.10 5.0 1.80.50 -- 3.69______________________________________ *HbF in μg/ml
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Verdict in Russian punk trial set for Aug. 17
Prosecutors and lawyers for church employees, who were described as the injured party in the case, insisted that they saw no political motives behind the band's actions, only blasphemy and hatred toward Orthodox believers.
"How did it happen that our performance, which was a small and clumsy stunt, brought so much trouble?" Alekhina told the court. "How can this happen in a healthy society? And now it takes thousands of people around the world to prove the obvious, to prove that the three of us are innocent."
Supporters in the courtroom greeted Alekhina's speech with enthusiastic applause, to which the judge responded, "This is not a theater."
About a dozen Pussy Riot supporters also gathered outside the courthouse, and at least three, including a protester wearing a balaclava, were detained, the Interfax news agency reported.
Before their church stunt, the band became an Internet sensation for performing a song that praised last winter's massive anti-Putin protests from a spot on Red Square used in the past for announcing czar's decrees. The group members have described themselves as feminists and accused the Russian leadership of infringing on the rights of women and the gay and lesbian community.
The Associated Press

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Doody's Review ServiceReviewer: Janice K. Warrington, MSNEd, RN (Delaware Technical & Community College)
Description: This is a textbook for those wishing to become certified nursing assistants.
Purpose: It provides basic, need-to-know information for individuals learning to practice the art of nursing as basic caregivers. This is certainly a worthy objective and the book provides basic information.
Audience: Nursing assistant students are the intended audience. Barbara Hegner, MS, RN, is former professor emeritus, Long Beach City College. Barbara Acello, MS, RN, an independent nurse consultant and educator, has worked in long-term care for more than 33 years and helped to write and develop mandatory state curricula for nurse aides and EMTs.
Features: The book provides detailed descriptions for the various procedures involved in providing basic nursing care — bed making, bathing, mobility, and so on. It includes unit reviews and testing material, detailed procedures, and text alerts on cultural aspects of care, critical information, and safety issues. The many color illustrations and pictures are, for the most part, relevant, but some are outdated. More importantly, in some cases, the book does not provide the most current evidence-based practice regarding procedures and the abbreviations list includes abbreviations that are no longer being used, according to Patient Safety Guidelines.
Assessment: This is a good basic care book. The next edition hopefully will include more up-to-date evidence-based practice and guidelines based on Patient Safety Guidelines.


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The Top Seven Accessory Trends for Fall 2011 with StyleSight, Day 4 It's the skins… Snakeskin, Python, Croc and Stingray … Snakeskin whether in print or genuine python, was clearly on almost every major designer's runway for Fall 2011 from Chloe, to Dries Van Noten to Prada, skin is back in. From classic boots to aviator…Published on September 06, 2011
More On: Dries Van Noten
Men's Style- Best of Men's Paris Lanvin, Dries Van Noten, Carven Collections for Spring Summer 2012
Runway Review: An edited look at cutting edge Paris men's fashion shows. Men's Style- Best of Men's Paris Lanvin, Dries Van Noten, Carven Collections for Spring Summer 2012. Khaki pants and polo shirt guys need not apply.There's a bounty of fashion-forward styles for guys who are not afraid to go the extra style mile.>> MORE: Fashion Shows and Runway ReviewsWhile the styling may be a bit risky for the everyday man, there certainly are some very clean and modern pieces that can easily be adapted to any cool guy's wardrobe.Take away the runway theatricality and there are plenty of looks to think about here. Our faves…Lanvin-Published on July 14, 2011
Carven, Dries Van Noten, fashion shows, Lanvin, Lucas Ossendrijve, men's style, Naveed Hussain, Paris, runway, runway review, Runway Trends & Reviews, spring, spring 2012, spring. summer
Men's Style- Best of Menswear Fall 2011 Runway, The Avant Gardes
Runway Review: Dries Van Noten, Jil Sander, Alexander McQueen, Marni, Miharayasuhiro.When it comes to men's style, there are two schools on the runway- the classics and the risk takers.We've shown you some of my favorite mens fashion shows from Europe from John Varvatos (the personal favorite for my hubby) , Louis Vuitton, Burberry Prorsum, Emporio Armani, Giorgio Armani, and even some super funk by way of Lady Gaga at Mugler.Then, there are the fashion risk takers. The men's fashion that has a bit more edge than the average guy can pull off, but plenty of style for those up to taking the chance! Naveed Hussain curates his favorite menswear fall 2011 collection from the catwalks of Paris and Milan for those guys wanting to take the fashion plunge! — S.H. After 9+ weeks of women's pre-fall came a month of menswear sandwiched in between the couture shows that just started the other day. The speed at which collections are being thrust upon fashion patrons now is head spinning to say the least. How can one remember what to buy, especially when the seasons deliveries aren't in sync with the reality of the weather at the time?>> MORE: Fashion Shows and Runway ReviewsPublished on January 26, 2011
Alexander McQueen, catwalk, Dries Van Noten, fall, FALL / WINTER 2011, fashion show, fashion show photos, fashion shows, Fashion Week, Fashion Weeks, Jil Sander, Marni, Men's Fashion Show Photos, men's style, menswear, Miharayasuhiro, Milan, Milan Fashion Week, Naveed Hussain, Paris, Paris Fashion Week, photos, Raf Simons, runway, Runway Trends & Reviews, Sarah Burton, VagbondNYC
Runway Review: Menswear, best of the classics fall 2010, Milan & Paris
"With the end of the last decade's heady maximalism behind us, and the dawn of a new era upon us with a loud call being heard for palette cleansing essentials in a time when the economy ain't so pretty, many are looking for easy, fuss-free propositions that are not only right for the times, but right for the future even.The day of instantly recognizable runway pieces is long gone and many venerable labels are looking back at not only their own heritages, but also the fundamentals of a smart wardrobe.>> See Runway Review: Menswear, best of cool for fall 2010The urban warrior, the svelte mod, the 80's electro playboy and the avant-garde techy futuro man we've seen many times around throughout the last decade have all taken a backseat (but that's not to say things are about to get too classic). In his place comes a clever use of house signatures for Fall/Winter 2010 that epitomize easy sophistication from institutions that you can always turn to for guaranteed quality.>> See Fall 2010 Fashion Show Photo GalleriesFour stand-out collections with "investment" potential & forward thinking clarity in this latest menswear movement were Giorgio Armani, Dries Van Noten, Lanvin & Prada."Published on January 28, 2010
Alber Elbaz, Armani, catwalk, classic, Dries Van Noten, fall, fall 2010, fashion shows, Fashion Week, Giorgio Armani, heritage, Lanvin, Lucas Ossendrijver, menswear, Milan, Naveed Hussain, Paris, Prada, runway, runway review, Runway Trends & Reviews, VagabondNYC


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You can now filter search results with text. Example: "person: john" will search only person pages for "john".
Welcome to Killville! Hope you survive the experience!...but probably you won't.
As if the Earth getting destroyed wasn't bad enough, this section of Battleworld has gone to hell...literally.
Natasha finally confronts the leader of CHAOS and does exactly what she needs to. In record time, in fact.
Character »
Red Ryan appears in 172 issues.
Member of the Challengers of the Unknown.
He is a member of the Sea Devils.
Jetsetter, maid of mystery.
After being murdered by the League of Assassins during a performance, circus acrobat Boston Brand was turned into a ghost by the Hindu goddess Rama Kushna. Becoming the superhero Deadman, Boston must aid the people he possesses to avoid eternity in Hell. He is currently part of Justice League Dark.
Honorary member of the Challengers of the Unknown
Walter "Prof" Haley is the leader of the Challengers of the Unknown team.
Amalgam version of Red Ryan and Human Torch
Kara Zor-El is Superman's cousin and last survivor of Krypton's Argo City. She has a brash and defiant personality that she developed in response to the destruction of Krypton. Currently, she is protecting Earth in Superman's stead at his request.
Botanist Alec Holland became the avatar of the Green, known as the Swamp Thing, following his death in a swamp as a result of a horrific accident. With the ability to control any form of plant life, Swamp Thing uses his powers to protect both the human and the plant worlds.
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Business Management and Economic Resources for Educators
To promote engagement between the educational and business communities, The Conference Board invites selected business school, college, and university presidents, deans, and faculty to become educational associate members.
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This membership can help you keep current on emerging business challenges and best practices, as well as the concerns of senior executives. Our issue-focused research — combined with our members' own experiences and the insights of our peer networks — promotes a fully informed, enterprise-wide perspective organized into three practice areas:
As an Educational Associate member, you'll benefit from research that is presented in a variety of formats. Here are some examples that have generated particular interest among executives in our member companies:
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Monrovia — The government of Liberia has instituted several measures, including prohibiting the gathering of people in the wake of the Ebola outbreak in the country, but it has been disclosed that in some counties cultural practices such as Sande bush where girls are gathered in one location, doing everything in common is still being practiced.
With Liberia now leading the number of cases and deaths ahead of both Guinea and Sierra Leone, rural dwellers are still running and operating Sande bushes that are considered traditional Bush Schools, in some countries, including the hometown of President Ellen Johnson Sirleaf, Bomi County, Minister of Gender and Development has told FrontPageAfrica.
Gender Minister Julia Duncan Cassell told FrontPageAfrica that the practice is dangerous especially with the ongoing Ebola crisis. "We have received reports that currently in Bomi County, there are Sande bushes being run. Bringing a group of girls together, put them at risk, because they do everything in common," said Minister Cassel.
Dangerous practice
Minister Cassel described such practice as dangerous and indicated that people are using the closure of schools as a means to gather the girls for such practice that exposes them to contracting the Ebola virus. Minister Cassell: "It is dangerous to run Sande bushes, and bring a group of people together. But I think people are taking advantage of the Ebola situation because schools are closed."
Minister Cassel noted that upon getting the news, she immediately informed the Minister of Internal Affairs Mr. Morris Dukuly, who informed his Assistant minister for culture to go and ensure that those Sande bushes are closed.
The Gender Minister observed that at this time parents should spend quality time with their children and get to know them instead of leaving them to be in such environment as Sande Bushes. "Parents should do things in their communities to keep the children busy. We are also asking that parents should engage their children and tell them stories of where they came from and who were their grandparents?
Use the opportunity while you are staying home so that your children can be protected. We are working with UNICEF to come up with some programs for the children to keep themselves busy and stimulate the minds while they are out of school", the Gender Minister told FPA.
Meima Sirleaf Karneh, Assistant Minister for Research and Technical Services at the Ministry of Gender, said during the first Ebola outbreak, the Ministry met with the female traditional Zoes educating them about the danger in clustering the children because the virus spreads through contact, and told them that by having the girls together in the bush, it is very dangerous at this time.
"After we did the awareness, we received information that there were Sande bushes ongoing in Bomi. And we reported the situation to the Internal Affairs through Mamatouma, who is the Chief Zoe, and she promised to have it investigated, because they were already told not to run the bush and put people at risk", Assistant Minister Karneh said
Mrs. Karneh said during their visitation in several communities, there were no Sande activities going; this she observed was due to the fact that normal schools were in section and traditional educational schools could not interrupt the formal schools.
Undermining Ebola fight
The Assistant Gender Minister described such practice as undermining the fight against the deadly virus. "I do not think people should take advantage of the Ebola situation and exploit it. Because running a Sande bush at this time, is undermining the process of fighting the Ebola virus. I think it is wrong and dangerous to do that, because when one person in the family gets it, everybody will get it because it is a silent killer", she warned.
Speaking about the importance of girls staying in school, Assistant Minister Karneh said in as much as they know traditional education is good, but in the modern world, one cannot easily succeed with it.
"When you take the child from the formal education and put her in the bush school, the school year is not waiting for them while their friends are going ahead and she is growing and left behind. At the end of the time, after she leaves the Sande, one old man would come and pay her bride price or dowry and she starts having children, and that is a violation of the girl's right," she said.
Sande activities ongoing in several counties
According to Minister Cassel, the practice is also being carried out in Lofa County where she disclosed that during a visit to the County by Vice President Joseph N. Boakai, he frowned on the practice after noticing they are taking place in one of two counties worse affected by the virus. Also in Grand Cape Mount County the practice is said to be taking place at the moment.
Bomi and Grand Cape Mount where the Sande practice is said to be taking place are amongst two of the counties quarantined by the Government to prevent further spread of the Ebola virus.
The Government of Liberia in 2012 suspended the practice of Sande from time indefinite and warned traditional leaders to adhere to the mandate. There has been widespread western and national condemnation of the aged old traditional practice which has existed in Liberia and other African countries for centuries.


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Press Releases | Returned : 21 results
The Dodgers will offer a total of seven promotions from June 4-10 for the club's sixth homestand of the season. Two theme nights will also take place. Tickets are available for all promotional dates listed below by calling 866-DODGERS or by visiting www.dodgers.com/promotions.
The Los Angeles Dodgers today acquired infielder Alberto Callaspo, left-handed reliever Ian Thomas, minor league right-handed pitcher Juan Jaime and left-handed pitcher Eric Stults from the Atlanta Braves in exchange for infielder Juan Uribe and right-handed pitcher Chris Withrow, who was on the 60-day disabled list. Callaspo will be in uniform for the Dodgers tonight, wearing No. 5, while Los Angeles optioned Thomas to Triple-A Oklahoma City, assigned Jaime to extended Spring Training in Arizona and designated Stults for assignment.
The Los Angeles Dodgers today recalled outfielder Chris Heisey from Triple-A Oklahoma City and designated right-handed pitcher Sergio Santos for assignment.
Hollywood Stars Night, one of the most popular promotional nights on the Los Angeles Dodgers schedule for many years, returns on Saturday, June 6 prior to the Dodgers taking on the St. Louis Cardinals.
The Los Angeles Dodgers today placed catcher Yasmani Grandal on the seven-day concussion disabled list and recalled catcher Austin Barnes (#65) from Triple-A Oklahoma City.
The Dodgers' fifth homestand of the season begins with a three-game series against the San Diego Padres tomorrow, May 22 (RHP Andrew Cashner vs. RHP Zack Greinke), Saturday, May 23 (RHP Ian Kennedy vs. RHP Mike Bolsinger) and Sunday, May 24 (RHP James Shields vs. RHP Carlos Frias).
The Dodgers announced today the third annual LGBT night at Dodger Stadium which will be held on Friday, June 19 (vs. SF, 7:10 p.m.). A special event ticket package that includes a game ticket for June 19 and an exclusive Dodger LGBT Night T-shirt (image attached) is available for purchase now exclusively by visiting www.dodgers.com/LGBT.
The Los Angeles Dodgers today announced the signings of Cuban infielder Hector Olivera to a six-year contract and Cuban right-handed pitcher Pablo Fernandez to a minor league deal. To make room for Olivera on the 40-man roster, Los Angeles designated left-handed pitcher Eury De La Rosa for assignment.
The Los Angeles Dodgers and the Chicago White Sox have teamed up to conduct the first-ever Dodgers-White Sox Baseball Fantasy Camp at Camelback Ranch, the teams' spring training facility in Glendale, Arizona. This authentic baseball experience for adults ages 21 years and older will be held January 18-24, 2016.
Today the Dodgers wish to thank their fans who purchased tickets to last night's game, played under heavy rain and wet conditions, with a special ticket offer for their patience and support. Starting now, 12:00 p.m., the Dodgers will offer a limited number of $20 Reserve Level tickets to games against the Braves on Memorial Day, Monday, May 25 (5:10 p.m.), Tuesday, May 26 (7:10 p.m.) and Wednesday, May 27 (7:10 p.m.)
The Los Angeles Dodgers today placed right-handed pitcher Pedro Baez on the 15-day disabled list (right pectoral strain) and recalled left-handed pitcher Daniel Coulombe from Triple-A Oklahoma City.
The Los Angeles Dodgers Foundation (LADF) announced today its 2015 Dodgers RBI Opening Day for the Los Angeles County Department of Parks and Recreation which will take place at Franklin D. Roosevelt Park, 7600 Graham Avenue in Los Angeles, Saturday, May 16 from 10:00 a.m. to 2:00 p.m. Dodger Manager Don Mattingly will attend the Dodgers RBI Opening Day and share brief remarks. Through his charity, Mattingly Charities, he will contribute batting gloves, metal bats and bat bags to this season's Dodgers RBI program. More than 400 participants are expected to attend and Dodger alumni Al Downing and Lee Lacy will partake.
The Dodgers' fourth homestand of the season concludes with a four-game series against the Colorado Rockies, Thursday, May 14 (RHP Chad Bettis vs. LHP Brett Anderson), Friday, May 15 (RHP Eddie Butler vs. LHP Clayton Kershaw), Saturday, May 16, (LHP Jorge De La Rosa vs. RHP Zack Greinke) and Sunday, May 17 (RHP Kyle Kendrick vs. RHP Mike Bolsinger). Games Thursday and Friday against the Rockies begin at 7:10 p.m. with auto gates opening at 4:40 p.m. and stadium gates opening at 5:10 p.m., Saturday's game is at 6:10 p.m. with auto gates opening at 3:30 p.m. and stadium gates at 3:30 p.m. and Sunday's contest starts at 1:10 p.m. with auto gates opening at 10:40 a.m. and stadium gates at 11:10 a.m. Tickets for all games against Colorado can be purchased by calling 866-DODGERS or by visiting www.dodgers.com/tickets.
The Los Angeles Dodgers today recalled right-handed pitcher Mike Bolsinger and optioned outfielder Chris Heisey to Triple-A Oklahoma City.
The Los Angeles Dodgers today announced the April winners of the Dodger Pride Awards, which are being classified into three categories: "Strong Mind Pitcher of the Month," "Strong Mind Position Player of the Month" and "Strong Mind Teammate of the Month." The winners of the pitcher and position player of the month are selected based on a strict statistical method incorporating several baseball measureables, including best first-pitch strike percentage, best 1-1 count strike percentage, best overall strike percentage, best walk-strikeout ratio, best swing percentage on strikes and lowest chase rate. The teammate of the month is voted on by players, coaches and staff.
The Dodgers' fourth homestand of the season begins with a three-game series against the Miami Marlins, Monday, May 11, Tuesday, May 12 and Wednesday, May 13. Contests on Monday and Tuesday against the Marlins take place at 7:10 p.m. with auto gates opening at 4:40 p.m. and stadium gates opening at 5:10 p.m. Wednesday's game begins at 4:50 p.m. with auto gates opening at 2:20 p.m. and stadium gates at 2:50 p.m.
The Los Angeles Dodgers today recalled outfielder Chris Heisey from Triple-A Oklahoma City and optioned right-handed pitcher Joe Wieland to Triple-A.
The Los Angeles Dodgers today recalled right-handed pitcher Joe Wieland from Triple-A Oklahoma City and optioned left-handed pitcher Daniel Coulombe to Triple-A.
Major League Baseball has announced the winners of the 2015 Honorary Bat Girl contest, which recognizes baseball fans who have been affected by breast cancer and who demonstrate a commitment to eradicating the disease. The Honorary Bat Girl for the Dodgers is Clara Gonzales of Los Angeles. Gonzales was diagnosed with stage III breast cancer in May 2014. Ten days after her diagnosis, she faced unemployment and lack of health insurance. Gonzales was able to receive support from the Komen Breast cancer society and began chemotherapy and radiation treatment. She is a life-long Dodger fan who wore Dodger blue and a Dodger wig during chemotherapy sessions and despite her illness, attends games regularly. Gonzales will throw the ceremonial first pitch prior to the 1:10 p.m. game against Colorado on May 17. She will also be presented with a customized jersey by Majestic Athletic and an engraved full-sized pink bat from Louisville Slugger.
The Los Angeles Dodgers today acquired right-handed pitcher Matt West from the Toronto Blue Jays in exchange for cash considerations and moved left-handed pitcher Hyun-Jin Ryu to the 60-day disabled list.
The Los Angeles Dodgers today acquired infielder Andy Wilkins from the Blue Jays in exchange for cash considerations and designated right-handed pitcher Scott Baker for assignment.


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Origo Metho 2B Recess Stove,4100
This Origo stove is equiped with non-pressurised alcohol burners.
Pot holders are available as a separate item, unit arrives as shown, with burner grid and 2 fuel canisters.
This unit is recessed into a bench top, with a fully enclosed base where the cannisters are held in place. A recess 150mm deep and 575 x 320 is required. Full top size is 620mm x 370mm
Unit should be mounted in a well ventilated area.
Click to View Product Manual (3,114kb) Click to View Product Manual (4.3Mb)
We have been selling this product, or an earlier model, for several years now. We usually stock at least a couple of these. If you order more than the 1 we have in stock, the delay is around 4 days to get more.


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Bigger trouble than shutdown looming
WASHINGTON President Barack Obama laid the blame for the government's partial shutdown at the feet of House Speaker John Boehner on Thursday, escalating a confrontation that is running the risk of a potentially damaging clash over the nation's borrowing authority.
The Treasury Department warned that a deadlock over raising the nation's debt limit could touch off a new recession even worse than the last one that Americans are still recovering from. Worry about prospects for resolving the debt question within the next two weeks deepened as the shutdown standoff dragged on.
The shutdown showdown grew more personal Thursday.
Speaking at a construction company in Washington's Maryland suburbs, Obama cast the House speaker as a captive of a small band of conservative Republicans who want to extract concessions in exchange for passing a short term spending bill that would restart the partially shuttered government.
"The only thing preventing people from going back to work and basic research starting back up and farmers and small business owners getting their loans, the only thing that is preventing all that from happening right now, today, in the next five minutes is that Speaker John Boehner won't even let the bill get a yes or no vote because he doesn't want to anger the extremists in his party," Obama said.
Boehner answered by batting blame back toward Obama and his "my-way-or-the-highway approach." Boehner said that if the president would negotiate to fix flaws in "Obamacare," the shutdown could end.
"The president's insistence on steamrolling ahead with this flawed program is irresponsible," said Boehner, R-Ohio.
Majority Leader Eric Cantor said the House would continue on its course of passing separate bills to remedy "situations that are in critical stages" because of the partial government shutdown that began Tuesday.
The House was expected to vote to for more money for National Guard and Reserves and for veterans programs during the day, and officials said legislation to help Head Start and possibly the WIC (Women, Infants and Children program) could soon be drafted, as well.
Senate Democrats made clear they will not agree to reopen the government on a piecemeal basis. "You can't fall for that legislative blackmail or it will get worse and worse and worse," said Sen. Chuck Schumer of New York.
Senate Democrats and Obama said the House must send them a measure that would restart all of government with no strings attached.
"Take a vote," Obama urged Boehner in his speech. "Stop this farce and end this shutdown right now."
It might not be so simple, however. Moderate Republicans have said they think they could provide enough votes to join with minority Democrats and push a bill through the House reopening the government with no restrictions on the health care law.
But under pressure from House GOP leaders, they failed to join Democratic efforts on Wednesday aimed at forcing the chamber to consider such legislation.
In the Senate, GOP Leader Mitch McConnell of Kentucky said the problem was "Democrats' refusal to apply simple fairness when it comes to Obamacare."


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Ground Weapon Scaling (size)
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Lt. Commander
Join Date: Dec 2007
Posts: 120
# 30
Originally Posted by
Sure you can state your opinion! That's fine; but when you do, please try and back it up with something constructive.
You merely stated that you think weapon scaling would be unrealistic. You said this without offering one reason why and seemingly ignoring the rest of the discussion that went on surrounding the topic you addressed.
Can you refute the reasons we have listed as to WHY weapon scaling would in-fact be realistic for the 25th century? More specifically can you refute why it would be even more realistic for the KDF with their varied species?
I apologize if I was terse, but it's hard to have a discussion with someone who does not bring anything to the table other than "I don't like it... It's unrealistic" and does not explain why.
With all due respect.
My point was not to make this game more unrealistic than necessary.
In any Star Trek series have you ever seen a Rifle, or other pices of equipment scaled to the person using it?
Not everyone carries his/her personal weapons all day around and weapons are usually stored in secured weapons stores or lockers on every starfleet ship.
Wouldnt it be a bit complicated to find everyone a fitting weapon every time the captain calls red alert?
Of course not, this equipment is standardised. I know some MK I weapons can be replicated, but most other weapons not.
How would you refuel them, would you reload them with bigger bullets, bigger energy cells, or the same as smaller versions have.
How comes that bigger versions do just the same damage as smaller version, althrough there is more room for (fill in some damage producing technobrabble stuff)?
No i wouldn't like to see scaling Weapon models they would just make the game even more unrealistic then necessary.
Live long and prosper.