WO2005085188A2

WO2005085188A2 - Compounds and methods for anti-tumor therapy

Info

Publication number: WO2005085188A2
Application number: PCT/US2005/006571
Authority: WO
Inventors: Karl J. Fisher; Matt Abelman
Original assignee: Compass Pharmaceuticals Llc
Priority date: 2004-03-02
Filing date: 2005-03-01
Publication date: 2005-09-15
Also published as: WO2005085188A3

Abstract

The present invention provides novel compounds and pharmaceutical compositions thereof, as well as methods for using the compounds and pharmaceutical compositions for treating tumors.

Description

Compounds and Methods for Anti-Tumor Therapy

Cross Reference This application claims the benefit of U.S. Provisional Patent Application Serial No. 60/549,243 filed March 2, 2004, which is hereby referenced in its entirety.

Field of the Invention The invention relates to the fields of chemistry, pharmaceuticals, and cancer.

Summary of the Invention The present invention provides novel compounds and pharmaceutical compositions thereof, as well as methods for using the compounds and pharmaceutical compositions for treating tumors.

Detailed Description of the Invention The present invention provides novel compounds and pharmaceutical compositions thereof, as well as methods for using the compounds and pharmaceutical compositions for treating tumors. Examples of specific tumor types that the compounds may be used to treat include, but are not limited to sarcomas, carcinomas (including but not limited to lung, renal cell, ovarian, melanoma, liver, bladder, and pancreatic carcinomas), and mesotheliomas. The compounds of the invention are disclosed below. These compounds can be administered individually or in combination, usually in the form of a pharmaceutical composition. Such compositions are prepared in a manner well known in the pharmaceutical art and comprise at least one active compound. Accordingly, a further aspect of the present invention also includes pharmaceutical compositions comprising as active ingredient compounds of the invention associated with a pharmaceutically

acceptable carrier. The compounds of the invention include pharmaceutically acceptable salts, esters, amides, and prodrugs therof, including but not limited to carboxylate salts, amino acid addition salts, esters, amides, and prodrugs of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention. The term "salts" refers to the relatively non-toxic, inorganic and organic acid addition salts of compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds or by separately reacting the purified compound in its free base form with a suitable organic or inorganic acid and isolating the salt thus formed. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts, and the like. These may include cations based on the alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations including, but not limited to ammonium, tetramethylammonium, tefraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. (See, for example, Berge S.M. et al., "Pharmaceutical Salts," J. Pharm. Sci, 1977;66:1-19 which is incorporated herein by reference.) Examples of pharmaceutically acceptable, non-toxic esters of the compounds of this invention include C^C_g alkyl esters, wherein the alkyl group is a straight or

branched, substituted or unsubstituted, C₅-G. cycloalkyl esters, as well as arylalkyl esters

such as benzyl and triphenylmethyl. C_j-C₄ alkyl esters are preferred, such as methyl,

ethyl, 2,2,2-trichloroethyl, and tert-butyl. Esters of the compounds of the present invention may be prepared according to conventional methods. Examples of pharmaceutically acceptable, non-toxic amides of the compounds of this invention include amides derived from ammonia, primary C_j-C₆ alkyl amines and

secondary C.-C₆ dialkyl amines, wherein the alkyl groups are straight or branched. In the

case of secondary amines, the amine may also be in the form of a 5- or 6-membered heterocycle containing one nitrogen atom. Amides derived from ammonia, C.-C₃ alkyl

primary amines and C.-C₂ dialkyl secondary amines are preferred. Amides of the

compounds of the invention may be prepared according to conventional methods. The term "prodrug" refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formulae, for example, by hydrolysis in blood. A thorough discussion of prodrugs is provided in T. Higuchi and N. Stella, "Pro-drugs as Novel Delivery Systems," Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are hereby incorporated by reference. hi a further aspect, the present invention provides methods for treating tumors comprising administering to a patient with a tumor an amount effective of one or more of the compounds of the invention to treat the tumor. Suitable tumors for treatment are as described above. As used herein, the term "effective amount" means a dosage sufficient to produce a desired result. The desired result can be subjective or objective improvement in the recipient of the dosage, a decrease in tumor size, time to progression of disease, and/or

survival. The compounds of the invention can be administered as the sole active pharmaceutical agent, or they can be used in combination with one or more other anti- tumor agents, including but not limited to chemotherapeutic agents. When administered as a combination, the therapeutic agents can be formulated as separate compositions that are given at the same time or different times, or the therapeutic agents can be given as a single composition. The compounds can further be administered in conjunction with other tumor treatments, including but not limited to radiation therapy and surgical removal of the tumor. The compounds may be made up in a solid form (including granules, powders or suppositories) or in a liquid form (e.g., solutions, suspensions, or emulsions). The compounds of the invention may be applied in a variety of solutions and may be subjected to conventional pharmaceutical operations such as sterilization and/or may contain conventional adjuvants, such as preservatives, stabilizers, wetting agents, emulsifiers, buffers etc. For administration, the compounds are ordinarily combined with one or more adjuvants appropriate for the indicated route of administration. The compounds may be admixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, stearic acid, talc, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, acacia, gelatin, sodium alginate, polyvinylpyrrolidine, and/or polyvinyl alcohol, and tableted or encapsulated for conventional administration. Alternatively, the compounds of this invention may be dissolved in saline, water, polyethylene glycol, propylene glycol, carboxyn etliyl cellulose colloidal solutions, ethanol, corn oil, peanut oil, cottonseed oil, sesame oil, tragacanth gum, and/or various buffers. Other adjuvants and modes of administration are well known in the pharmaceutical art. The carrier or diluent may include time delay material, such as glyceryl monostearate or glyceryl distearate alone or with a wax, or other materials well known in the art. Pharmaceutical compositions containing the compounds described herein are administered to an individual having a tumor. In therapeutic applications, compositions are administered to a human patient in an amount sufficient to cause regression of the tumor, or at least partially arrest tumorigenesis and metastasis. Amounts effective for this use depend on factors including, but not limited to, the nature of the compound (specific activity, etc.), the manner of administration, the stage and severity of the cancer, the weight and general state of health of the patient, and the judgment of the prescribing physician. The active compounds are effective over a wide dosage range. For example,

dosages per day will normally fall within the range of about 1 μg/kg body weight to about

100 mg/kg of body weight. It will be understood that the amount of the compound actually administered will be determined by a physician, in the light of the relevant circumstances including the condition to be treated, the choice of compound to be administered, the chosen route of administration, the age, weight, and response of the individual patient, disorders affecting the heart, and other specific organ dysfunction, and therefore the above dosage ranges are not intended to limit the scope of the invention in any way. The compounds of the invention may be administered by any suitable route, including orally, parentally, by inhalation or rectally in dosage unit formulations containing conventional pharmaceutically acceptable carriers, adjuvants, and vehicles, including liposomes. The term parenteral as used herein includes, subcutaneous, intravenous, intraarterial, intramuscular, intrasternal, intratendinous, intraspinal, intracranial, intrathoracic, infusion techniques, intracavity, or intraperitoneally. In yet further aspects, the invention provides an article of manufacture comprising packaging material and the above pharmaceutical compositions. The instant invention may be embodied in other forms or carried out in other ways without departing from the spirit or essential characteristics thereof. The present disclosure and enumerated examples are therefore to be considered as in all respects illustrative and not restrictive, and all equivalency are intended to be embraced therein. One of ordinary skill in the art would be able to recognize equivalent embodiments of the instant invention, and be able to practice such embodiments using the teacliing of the instant disclosure and only routine experimentation.

Examples

Example 1. Tissue Processing Excess tissue specimens obtained from organs and tissues such as lung and testicle were obtained freshly at the time of surgery and samples were sent for pathological testing. For diagnosis and grading of tissue samples (ie: prior to processing), hematoxylin and eosin stained tissue sections were examined by a pathologist. If the diagnosis and grading of the tissue concurred with the determination made by the surgical pathologist that provided the tissue, then the tissue was used in the screen. If there was no agreement, then two additional pathologists served as referees. If no consensus was reached, then the tissue was discarded. The remaining tissue was used to prepare cell suspensions. The tissue was initially treated enzymatically via standard methods until only undigested material remained. The digested cell suspension was filtered through one or more screens of between 40 micron and 100 micron porosity. The resulting cell suspension was further purified via isokinetic density centrifugation. Additional normal cells were removed from the cell suspension by negative immunoselection with a combination of monoclonal antibodies linked to magnetic beads (Dynal) that were used according to the manufacturers' instructions. The remaining cells were placed into appropriate medium, frozen down in 1.0 mL aliquots, and stored until use.

Example 2. General Screen/Bioassay Procedures After tissue processing, the relative purity of the resulting cell suspension was determined by cytological examination after pap staining. Only those cell preparations greater than 80% tumor cells were used for testing of candidate compounds. If there was any doubt about the percentage of tumor cells in the cell preparation, additional pathologists served as referees to make a determination. Cell preparations that passed histological and cytological examination for diagnosis, grading, and cell purity were thawed at 37°C and resuspended in tissue culture ^• medium designed to maintain the cells during the incubation period. The live and dead cells were counted and the cells were diluted in culture medium to 1.0 x 10³ live cells/test well for tumor cells and 3.3 x 10³ live cells/test well for normal cells. The cells were added to microtiter plates and incubated at 37°C overnight with 10 μM of the candidate compounds that were added at 1/10th the volume of the cell suspension. Alamar Blue (Accumed International, Westlake OH) was then added to the cells at 1/10 the volume of the well, and the cells were further incubated at 37°C for various times. Alamar Blue dye measures cellular re-dox reactions (ie: cellular respiration) whereby a spectral shift occurs upon reduction of the dye. (Excitation 530

nm; emission 590 nm) The kinetics of cellular re-dox reactions were subsequently measured at various times, for example at 3 hours, 3 days, and 5 days post-dye addition. These measurements, in comparison with control cells (untreated with compound) and media controls (test wells without cells) provide the percent inhibition of cellular mitochondrial respiration as a result of candidate compound treatment, as well as IC₅o determinations. The Alamar Blue data were subsequently confirmed by microscopic observation, and by the use of calcein AM (Molecular Probes, Eugene OR), a cell permeant esterase substrate that measures both esterase activity and cell membrane activity. If the cell is alive, the dye is converted into a fluorogenic substrate by intracellular esterases and is retained by the cell (excitation 485 nm; emission 530 nm). If the cells are dead, the calcein AM rapidly leaks from the cells and is not converted into a fluorogenic substrate. Thus, the assay is useful for cytotoxicity testing.

Example 3. Anti-Tumor Screen In a blinded fashion, approximately 10,000 compounds were tested at a rate of 1,000-4,000 compounds per run set against colon, lung, and sarcoma tumors. The anti- tumor screen utilized was composed of three tiers as follows. In screen 1, patient tumor cells were tested in singles, with candidate compounds at a concentration of 10 μM.

Samples that showed at least 80% inhibition (compared to cell and media controls) and/or two standard deviations from the mean of the plate samples were advanced. In the first part of the second test (screen 2a), the compounds were re-tested, in duplicate, at 10 μM concentrations on patient tumors. Compounds that re-confirmed were then tested, in duplicate, at 10 μM concentration on patient normal cells. Samples that exhibited at least 80% inhibition on tumor cells and no more than 20% inhibition of normal cells were tested in the second part (screen 2b). Using the screen disclosed above, the compounds disclosed in Figures 1 and 2 were demonstrated to have anti-tumor activity. These data clearly show that the compounds of the invention can be used as an anti-tumor agent against sarcomas, mesotheliomas, and a wide range of carcinomas.

Claims

We claim:

1. A compound selected from the group consisting of those compounds disclosed in Figures 1 and 2, or salts thtereof.

2. A pharmaceutical composition comprising one or more compounds selected from those disclosed in Figures 1 and 2, or salts thereof, and a pharmaceutically acceptable carrier.

3. Use of a compound according to claim 1 for the preparation of a medicament to treat a tumor.

4. Use of a pharmaceutical composition according to claim 2 for the preparation of a medicament to treat a tumor.