Compositions and methods for reducing cell viability

This invention is directed to compositions, methods and kits that can be used for reducing cell viability, inducing cell apoptosis, and inhibiting cell proliferation, such as for the treatment of cancer. The invention is also directed to compositions, methods and kits that can be used for treating inflammatory conditions, such as by modulating inflammatory cytokines.

All patents, patent applications and publications cited herein are hereby incorporated by reference in their entirety. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described and claimed herein.

FIELD OF THE INVENTION

This invention is directed to compositions, methods and kits that can be used for reducing cell viability, inducing cell apoptosis, and inhibiting cell proliferation, such as for the treatment of cancer. The invention is also directed to compositions, methods and kits that can be used for treating inflammatory conditions, such as by modulating inflammatory cytokines.

BACKGROUND OF THE INVENTION

Cyclooxygenase (COX) is the key enzyme of arachidonic acid metabolism. Arachidonic acid metabolite, generated by COX, is implicated in cancer pathogenesis, for example the colorectal cancer (CRC) pathogenesis. The development of anti-cancer drugs targeting COX-2 has been limited by unwanted toxic side effects.

SUMMARY OF THE INVENTION

The invention provides for compounds for treating cancer.

Aspects of the invention are directed towards a compound of Formula I:

In an embodiment, the aryl is phenyl.

In an embodiment, the compound is:

In an embodiment, the compound is:

Aspects of the invention are directed towards a compound of Formula II:

In an embodiment, the aryl is phenyl.

In an embodiment, the compound is:

In an embodiment, the compound is:

Aspects of the invention are further directed towards methods of reducing cell viability comprising contacting a cell with an effective amount of a compound described herein.

Aspects of the invention are still further directed towards methods of inducing apoptosis of a cell comprising contacting a cell with an effective amount of a compound described herein.

Aspects of the invention are also directed towards methods of inhibiting cell proliferation comprising contacting a cell with an effective amount of a compound described herein.

Still further, aspects of the invention are directed towards methods of regulating epithelial-mesenchymal transition (EMT) comprising contacting a cell with an effective amount of a compound described herein.

Yet further, aspects of the invention are directed towards methods of inhibiting tumor growth comprising contacting a cell with an effective amount of a compound described herein.

In embodiments, the compound comprises

In embodiments, the compound comprises

Embodiments comprise a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable carrier.

Embodiments can further comprise a second active agent, for example a non-steroidal anti-inflammatory drug (NSAID), a selective COX-2 inhibitor, a non-selective COX inhibitor, or a combination thereof. Non-limiting examples of the selective COX-2 inhibitor comprises celecoxib, refecoxib, parecoxib and valdecoxib.

Aspects of the invention are directed towards methods of reducing cell viability comprising contacting a cell with an effective amount of a compound of Formula IV

Aspects of the invention are directed towards methods of inducing apoptosis of a cell comprising contacting a cell with an effective amount of a compound of Formula IV:

In embodiments, the compound modulates levels of survivin, Bcl-2, and Bax.

Aspects of the invention are directed towards methods of inhibiting cell proliferation comprising contacting a cell with an effective amount of a compound of Formula IV:

In embodiments, cell proliferation is indicated by measuring TCF/LEF1 activity.

Aspects of the invention are directed towards methods of regulating epithelial-mesenchymal transition (EMT) comprising contacting a cell with an effective amount of a compound of Formula IV:

Aspects of the invention are directed towards methods of inhibiting tumor growth comprising contacting a cell with an effective amount of a compound of Formula IV:

In embodiments, tumor growth is inhibited by inducing cellular apoptosis and/or inhibiting cellular proliferation.

In embodiments, the aryl group is phenyl.

In embodiments, the compound comprises the following chemical structure:

In embodiments, the compound comprises the following chemical structure:

In embodiments, the compound comprises the following chemical structure:

In embodiments, the cell comprises a cancer cell, such as a colorectal/colon cancer cell, pancreatic cancer cell, or lung cancer cell.

Aspects of the invention are directed towards methods of suppressing an inflammatory response comprising contacting a cell with an effective amount of a compound described herein.

Aspects of the invention are directed towards methods of modulating an inflammatory cytokine in a cell comprising contacting a cell with an effective amount of a compound described herein. For example, the cytokine comprises a pro-inflammatory cytokine (IL-6, IL-8, TNFα, IL-1β), an anti-inflammatory cytokine (IL4), and/or a chemokine (CCL2).

In embodiments, the compound is administered to a subject prior to contacting.

In embodiments, the compound is administered in a single dose.

In embodiments, the compound is administered continuously.

In embodiments, the compound is administered at intervals of about 4 hours, 12 hours, or 24 hours.

In embodiments, the compound is administered orally, parentally, transdermally, or nasally.

In embodiments, the compound exhibits anti-cancer effect while the risk of toxicity is reduced.

In embodiments, the compound exhibits anti-inflammatory effect while the risk of toxicity is reduced

Embodiments can further comprise measuring serum biomarkers indicative of cardiac function, stomach function, liver function, kidney function, or a combination thereof. Non-limiting examples of biomarkers indicative of cardiac function comprise serum cardiac troponin-I (cTnI), serum creatine kinase-MB (CK-MB), or a combination thereof. Non-limiting examples of biomarkers indicative of liver function and/or kidney function comprise serum enzymes, total protein, and total albumin.

Aspects of the invention are also directed towards a medical kit suitable for the treatment of cancer, comprising printed instructions for administering a compound described herein to a subject afflicted with a cancer; or a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable carrier. In embodiments, the compound is a selective, COX-2 inhibitor.

Aspects of the invention are also directed towards methods of treating a neurological disorder comprising administering to a subject afflicted with a neurological disorder a therapeutically effective amount of a compound described herein.

Other objects and advantages of this invention will become readily apparent from the ensuing description.

DETAILED DESCRIPTION OF THE INVENTION

Abbreviations and Definitions

The singular forms “a”, “an” and “the” include plural reference unless the context clearly dictates otherwise. The use of the word “a” or “an” when used in conjunction with the term “comprising” in the claims and/or the specification may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.”

Wherever any of the phrases “for example,” “such as,” “including” and the like are used herein, the phrase “and without limitation” is understood to follow unless explicitly stated otherwise. Similarly “an example,” “exemplary” and the like are understood to be nonlimiting.

The term “substantially” allows for deviations from the descriptor that do not negatively impact the intended purpose. Descriptive terms are understood to be modified by the term “substantially” even if the word “substantially” is not explicitly recited.

The terms “comprising” and “including” and “having” and “involving” (and similarly “comprises”, “includes,” “has,” and “involves”) and the like are used interchangeably and have the same meaning. Specifically, each of the terms is defined consistent with the common United States patent law definition of “comprising” and is therefore interpreted to be an open term meaning “at least the following,” and is also interpreted not to exclude additional features, limitations, aspects, etc. Thus, for example, “a process involving steps a, b, and c” means that the process includes at least steps a, b and c. Wherever the terms “a” or “an” are used, “one or more” is understood, unless such interpretation is nonsensical in context.

Cyclooxygenase (COX) is the key enzyme of arachidonic acid metabolism. Arachidonic acid metabolite, generated by COX, is implicated in cancer pathogenesis, for example the colorectal cancer (CRC) pathogenesis. Multiple lines of evidence indicate that cyclooxygenase-2 (COX-2) is a bonafide pharmacological target for anticancer therapy; however, subsequent development has been limited by unwanted toxic side effects, such as cardiotoxicity and gastric injury. See, for example, Casanova I, Parreño M, Farré L, Guerrero S, Céspedes M V, Pavon M A, Sancho F J, Marcuello E, Trias M, Mangues R Int J Cancer. 2006, 118 (10), 2381-9.

Epidemiological studies show that use of non-steroidal anti-inflammatory drugs (NSAIDs), prototypic inhibitors of COX, is associated with a reduced risk of several malignancies, including colorectal cancer. NSAIDs blockcellular proliferation and induce apoptosis in malignant cells and thereby inhibit tumor growth. Further, several studies have demonstrated that tumor formation and growth are reduced in animals that are either engineered to be COX-2 deficient or treated with a selective COX-2 inhibitor. In clinical trial, treatment with celecoxib, a selective COX-2 inhibitor, reduced the number of colorectal polyps in patients with familial adenomatous polyposis (FAP).

In recent years, several selective COX-2 inhibitors were designed to overcome the problems of toxicity, such as cardiotoxicity and gastric injury (associated with classical NSAIDs). However, most of these drugs were voluntary withdrawn from the market due to cardiovascular toxicity and their effects on gastric injury. Therefore, there is an unmet need to develop effective anti-inflammatory agents with anticancer properties that are non-toxic or with reduced toxicity. Aspects of the invention are directed towards the design, synthesis, characterization and preclinical and clinals uses of novel Prostaglandin Endoperoxide Synthetase-2 (PTGS-2, gene encoding for COX-2) inhibitors or COX-2 inhibitors for the prevention and/or treatment of inflammatory diseases and cancer which are non-toxic or demonstrate reduced toxicity, and which are safe for ulcerogenic, hepatic, and renal funcations and cardiotoxic liabilities.

Aspects of the invention are directed towards compounds that exhibit anti-cancer and/or anti-inflammatory effects which the risk of toxicity is reduced. For example, the compounds can exhibit reduced cardiotoxicity, gastrotoxicity, hepatotoxicity, nephrotoxicity, or any combination thereof.

Aspects of the invention are directed towards a compound of Formula I:

For example, the compound is:

In embodiments, the aryl is phenyl, for example unsubstituted phenyl.

Aspects of the invention are directed towards a compound of Formula II:

Where in

R2is an aryl; and

For example, the compound is:

In embodiments, the aryl is phenyl, for example unsubstituted phenyl.

Aspects of the invention are further directed towards a compound of Formula III:

For example, the compound is

In embodiments, the aryl group (Ar), for example, naphthyl or phenyl non-limiting examples of which comprise 4-Methylphenyl, 4-Methoxyphenyl, 4-Chlorophenyl, or 4-Nitrophenyl.

In embodiments, the compound is a selective COX-2 inhibitor.

Aspects of the invention are further directed towards a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable carrier.

In embodiments, the pharmaceutical composition further comprises a second active agent. For example, the second active agent comprises a non-steroidal anti-inflammatory drug (NSAID), a selective COX-2 inhibitor (such as celecoxib, refecoxib, parecoxib and valdecoxib), a non-selective COX inhibitor, or a combination thereof.

Embodiments of the invention demonstrate to have no or reduced levels of toxicity relative to known selective COX-2 inhibitors, such as cardiotoxicity or gastric injury. “Toxicity” can refer to any adverse and/or undesired effect of a compound or pharmaceutical composition on the metabolism or functioning of a cell, tissue, organ or body part, or subject. The amount of toxicity associated with a compound or composition can vary with several conditions including, but not limited to, the amount of compound or composition present, the components present in the composition, the formulation of the composition, the environmental conditions and physiological state of the cell, tissue, organ or body part, or subject, etc. For example, toxicity can refer to cardiotoxicity, gastrotoxicity, hepatotoxicity, or nephrotoxicity.

“Cardiac toxicity,” which can also be referred to as “cardiotoxicity,” can refer to any toxic side effect on the cardiac system, and can be caused by a medicinal agent administered for the purpose of treating a medical disorder. Cardiac toxicity can be evaluated, for example, based on any one or more of: incidence of symptomatic left ventricular systolic dysfunction (LVSD) or congestive heart failure (CHF), or decrease in left ventricular ejection fraction (LVEF).

“Gastric toxicity,” which can also be referred to as “gastrotoxicity,” can refer to any toxic side effect on the stomach, and can be caused by a medicinal agent administered for the purpose of treating a medical disorder. Gastrotoxicity can be evaluated, for example, based on any one or more of ulcerogenic liability study. See, for example, I. Szelenyl, K. Thiemer, Distention ulcer as a model for testing of drugs for ulcerogenic side effects, Arch. Toxicol. 41 (1978) 99-105.

The term “hepatotoxicity” can refer to the chemical- or drug-induced liver damage. Drug-induced liver injury or damage is a cause of acute or chronic liver disease. Hepatotoxicity can be caused by certain medicinal agents, when taken in overdoses or sometimes even when introduced within therapeutic ranges. Hepatotoxicity can be evaluated, for example, based on any one or more of liver function tests, such as serum biochemical analysis (A. G. Banerjee, N. Das, S. A. Shengule, R. S. Srivastava and S. K. Shrivastava, Eur J Med Chem. 2015, 101, 81-95) of serum glutamic oxaloacetate transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase reflecting hepatic functions; or histopathological assessment of liver (A. E. Galigher, E. N. Kozloff, second ed., in: Lea, Febiger (Eds.), Essentials of Practical Microtechnique, vol. 20, Hagerstown Publications, Maryland, USA, 1971, p. 77).

The term “nephrotoxicity” can refer to a condition in which impairment of renal function occurs, and can be caused by a medicinal agent administered for the purpose of treating a medical disorder. Nephrotoxicity can be evaluated, for example, based on any one or more of kidney function tests, such as serum biochemical analysis (e.g., albumin, creatinine and Blood Urea Nitrogen; see for example, A. G. Banerjee, N. Das, S. A. Shengule, R. S. Srivastava and S. K. Shrivastava, Eur J Med Chem. 2015, 101, 81-95); or histopathological assessment of kidney (see, for example, E. Galigher, E. N. Kozloff, second ed., in: Lea, Febiger (Eds.), Essentials of Practical Microtechnique, vol. 20, Hagerstown Publications, Maryland, USA, 1971, p. 77)

In some embodiments, the compound or composition comprising the same has a reduced risk of toxicity, for example when compared to known COX-2 inhibitors such as NSAIDs, when administered to a subject in vivo. For example, the composition can reduce the risk of toxicity by at least about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50%.

The log P value of a compound is a measure of the compound's hydrophilicity. All the synthesized compounds were evaluated for their experimental log P values using shake-flask method. See, for example, S. O. Podunavac-kuzmanovic, D. D. Cvetkovic, D. J. Barna, The effect of lipophilicity on the antibacterial activity of some 1-benzylbenzimidazole derivatives, J. Serb. Chem. Soc. 73 (2008) 967-978.

Cancer

Aspects of the invention are directed towards compositions and methods for the treatment of cancer. “Cancer” refers to a general term for diseases caused by any type of tumor, such as a malignant tumor. Malignant, as applies to tumors, refers to tumors resulting from abnormal uncontrolled growth of cells and includes, but is not limited to, colorectal cancer, colon cancer, pancreatic cancer, or lung cancer.

Thus, compounds of the invention can be considered “anti-cancer agents” (also referred to as anti-neoplastic agents or anti-tumor agents), which can refer to any agent used in the treatment of cancer or neoplasm. Anti-cancer agents, when used alone or in combination with other compounds, can alleviate, reduce, ameliorate, prevent, or place or maintain in a state of remission of clinical symptoms or diagnostic markers associated with neoplasm, tumor or cancer, and can be used in methods, combinations and compositions provided herein.

Neurological Disorders

Aspects of the invention are directed towards compositions and methods for the treatment of neurological disorders. “Neurological disorder” or “neurological disease” can refer to a general term for diseases of the brain, spine and the nerves that connect them. Non-limiting examples of diseases of the nervous system comprise brain tumors, epilepsy, Parkinson's disease, stroke, and frontotemporal dementia.

Thus, compounds of the invention can be used in the treatment of a neurological disorder, and when used alone or in combination with other compounds, can alleviate, reduce, ameliorate, prevent, or place or maintain in a state of remission of clinical symptoms or diagnostic markers associated with the neurological disorder, and can be used in methods, combinations and compositions provided herein.

Pharmaceutical Combinations

Aspects of the invention are directed towards compositions and methods for the treatment of cancer and inflammatory conditions. Embodiments can comprise selective COX-2 inhibitors as described herein, and can be a component of pharmaceutical combinations for the treatment of cellular proliferative diseases (such as cancer) and/or inflammatory conditions.

The pharmaceutical combinations of the present invention comprise compounds as described herein, such as compounds of Formula I, II and III or IV, in an admixture along with a pharmaceutically acceptable carrier prepared according to conventional pharmaceutical techniques. According to the invention, a pharmaceutically acceptable carrier can comprise any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Non-limiting examples of pharmaceutically acceptable carriers comprise solid or liquid fillers, diluents, and encapsulating substances, including but not limited tolactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starches, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl benzoate, propyl benzoate, talc, magnesium stearate, and mineral oil. The amount of the carrier employed in conjunction with the combination is sufficient to provide a practical quantity of material per unit dose of the compound.

The use of such media and agents for pharmaceutically active substances is well known in the art. Any conventional media or agent that is compatible with the active compound can be used. Supplementary active compounds can also be incorporated into the compositions.

Various oral dosages forms can be employed, non-limiting examples of which comprise solid forms such as tablets, capsules, granules, suppositories and/or powders. Tablets can be compressed, tablet triturates, enteric-coated, sugar-coated, film-coated or multiple compressed, containing suitable binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents. Liquid oral dosage forms comprise aqueous solutions, emulsions, suspensions, syrups, aerosols and/or reconstituted solutions and/or suspensions. The composition may alternatively be formulated for external topical application, or in the form of a sterile injectable solution.

The present invention also comprises the formation of pharmaceutically acceptable, stable salts of the compounds as described herein with metals or amines. Non-limiting examples of metals used as cations comprise alkali metals such as Na+or K+and alkaline-earth metals such as Mg2+and Ca2+. Non-limiting examples of amines comprise N,N-dibenzylethylenediamine, chloro-procaine, choline, diethanolamine, ethylenediamine, N-methylglucamine and procaine.

As an exemplary embodiment, pharmaceutical combinations of the invention can be administered orally, either in the form of tablets containing excipients such as starch or lactose, or in capsules, either alone or mixed with excipients, or in the form of syrups or suspensions containing coloring or flavoring agents. They can also be injected parenterally, for example intramuscularly, intravenously or subcutaneously. In parenteral administration, they can be used in the form of a sterile aqueous solution which can contain other solutes, such as, for example, any salt or glucose in order to make the solution isotonic.

The compounds of the present invention can be administered to a subject for the treatment of cancer or an inflammatory condition, for example orally, either covered in gelatin capsules or compressed in lozenges. For oral therapeutic administration, said compounds can be mixed with excipients and used in the form of lozenges, tablets, capsules, elixirs, suspensions, syrups, wafers, chewing gum, and the like. These preparations could contain at least 0.5% of active compound, but can vary depending on each form, in particular between 4% and 75% approximately of the weight of each unit. The amount of active compound in such compositions should be that which is necessary for obtaining the corresponding dosage. For example, the compositions and preparations as described herein can be prepared in such a way that each oral dosage unit can contain between 0.1 mg and 300 mg of the active compound.

In parenteral therapeutic administration, the active compounds of this invention can be incorporated in a solution or suspension. Such preparations, for example, can contain at least 0.1% of the active compound, but can vary between 0.5% and 50% approximately of the weight of the preparation. For example, such preparations comprise about 0.1%, 0.5%, 1%, 5%, 10%, 15%, 25%, 30%, 35%, 40%, 45%, 50%, of the weight of the preparation. The amount of active compound in such compositions should be that which is necessary for obtaining the corresponding dosage. The compositions and preparations as described herein can be prepared in such a way that each parenteral dosage unit can contain between 0.01 mg and 1000 mg, for example between about 0.5 mg and 100 mg of the active compound, for example. While intramuscular administration can be given in a single dose or divided into up to multiple doses, such as three doses, intravenous administration can include a drip device for giving the dose by venoclysis. Parenteral administration can be performed by means of ampoules, disposable syringes or multiple-dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use can include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers can include physiological saline, bacteriostatic water, Cremophor EM™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In embodiments, the composition can be sterile and should be fluid to the extent that easy syringability exists. It can be stable under the conditions of manufacture and storage and can be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, a pharmaceutically acceptable polyol like glycerol, propylene glycol, liquid polyetheylene glycol, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, and thimerosal. In many cases, it can be useful to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can occur by including an agent in the composition which delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated herein, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated herein. In the case of sterile powders for the preparation of sterile injectable solutions, examples of useful preparation methods are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed.

Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

The compounds of the present invention can be administered to a subject in a single dose for the treatment of cancer or an inflammatory condition, or as multiple doses over a period of time. Further, the compound can be administered at intervals of about 4 hours, 8 hours, 12 hours, 24 hours, or longer. In embodiments, the compounds of the invention can be administered continuously over a period of time, such as for 4 hours, 8 hours, 12 hours, 24 hours, or longer.

Of necessity, there will be variations which will depend on the weight and conditions of the subject to be treated and on the particular administration route selected.

Methods of Treatment

Embodiments can be used for the treatment of cancer or an inflammatory condition. “Treatment” can refer to an approach for obtaining beneficial or desired clinical results. For example, the term “treating cancer” or “treatment of cancer” can refer to administration of a compound to a subject afflicted with a cancerous condition alleviates the cancerous condition by killing the cancerous cells, but also to an effect that results in the inhibition of growth and/or metastasis of the cancer. Non-limiting examples of cancers that can be treated by aspects of the invention comprise colorectal cancer, colon cancer, pancreatic cancer, lung cancer, and the like.

Aspects of the invention are directed towards a method of reducing cell viability by contacting a cell, such as a cancer cell, with an effective amount of a compound as described herein.

Aspects of the invention are further directed towards a method of inducing apoptosis of a cell, such as a cancer cell, by contacting a cell with an effective amount of a compound as described herein. In embodiments, apoptosis comprises caspase-dependent apoptosis, such as apoptosis dependent upon caspase 3 and/or caspase 7. In other embodiments, the compound can induce apoptosis by modulating levels of apoptosis-regulatory proteins, such as survivin, Bcl-2, and Bax.

Aspects of the invention are still further directed towards a method of inhibiting cell proliferation of a cell, such as a cancer cell, by contacting a cell with an effective amount of a compound as described herein. Cell proliferation can be indicated, for example, by measuring TCF/LEF1 activity.

Aspects of the invention are also directed towards method of regulating epithelial-mesenchymal transition (EMT) by contacting a cell, such as a cancer cell, with an effective amount of a compound described herein.

Further, aspects of the invention are directed towards a method of inhibiting growth of a cancer cell or tumor by contacting a cell, such as a cancer cell, with an effective amount of a compound described herein. For example, tumor growth can be inhibited by inducing cellular apoptosis and/or inhibiting cellular proliferation.

In embodiments, the compound is a compound of Formula I or II. For example, the compound is:

In other examples, the compound is:

In embodiments, the compound is a compound of Formula IV:

R5is an aryl group; and

R6is O or S.

For example, the compound is:

In other examples, the compound is:

In still other examples, the compound is:

As described herein, the aryl group is a napthyl or a phenyl, non-limiting examples of which comprise 4-Methylphenyl, 4-Methoxyphenyl, 4-Chlorophenyl, or 4-Nitrophenyl.

Still further, aspects of the invention are directed towards methods of suppressing an inflammatory response comprising contacting a cell with an effective amount of a compound as described herein.

In embodiments, aspects are directed towards methods of modulating inflammatory cytokines (i.e., cytokines that regulate the inflammatory response) in a cell comprising contacting a cell with an effective amount of a compound as described herein. Non-limiting examples of such inflammatory cytokines comprise pro-inflammatory cytokine (such as IL-6, IL-8, TNFα, IL-1β), anti-inflammatory cytokine (such as IL4), and/or chemokines (such as CCL2).

Aspects of the invention are directed towards methods of administering to a subject compounds that exhibit anti-cancer and/or anti-inflammatory effects while the risk of toxicity is reduced. For example, the compounds can exhibit reduced cardiotoxicity, gastrotoxicity, hepatotoxicity, nephrotoxicity, or any combination thereof. Embodiments can further comprise measuring serum biomarkers to determine toxicity. For example, embodiments can further comprise measuring biomarkers indicative of cardiac function (such as serum cardiac troponin-I (cTnI), serum creatine kinase-MB (CK-MB), or a combination thereof), stomach function, liver function and/or kidney function (such as serum enzymes, total protein, and total albumin), or any combination thereof.

In embodiments, a compound as described herein is used as the only physically active compound in the treatment of cancer without a second active agent.

In other embodiments, the compositions as described herein can be administered to a subject concurrently with and/or in combination with a second active ingredient, such as a non-steroidal anti-inflammatory drug (NSAID), a selective COX-2 inhibitor, a non-selective COX inhibitor, or a combination thereof. Nonsteroidal anti-inflammatory drugs (NSAIDs) block the COX enzymes and reduce prostaglandins throughout the body. As a consequence, ongoing inflammation is reduced. Non-limiting examples of NSAIDs comprise aspirin, celecoxib, diclofenac, diflunisal, etodolac, ibuprofen, indomethacin, ketoprofen, ketorolac, nabumetone, naproxen, oxaprozin, piroxicam, salsalate, sulindac, and tolmetin.

In embodiments, the compound can be administered to a subject prior to contacting a cell. Compounds of as described herein can be incorporated into pharmaceutical compositions suitable for administration to a subject in need thereof. Such compositions can comprise a compound as described herein and a pharmaceutically acceptable carrier. Thus, in some embodiments, the compounds of the invention are present in a pharmaceutical composition.

For example, a pharmaceutical compositions comprising a compound as described herein can be used for treating cancer, such as a therapeutically effective amount of a compound as described herein in admixture with a pharmaceutical acceptable carrier or excipient. For example, a therapeutically effective amount the compound can be administered to a subject in need thereof so as to reduce the size of a tumor, or to prevent the tumor from growing.

Embodiments comprise administering to a subject an effective amount of a composition as described hereinfor the treatment of cancer or inflammatory conditions.

An “effective amount”, “sufficient amount” or “therapeutically effective amount” can refer to an amount sufficient to effect beneficial or desired clinical result, such as killing the cancerous cells, inhibiting the growth of the cancer, inhibiting the metastasis of the cancer, and/or reducing or inhibiting inflammation.

Specific compositions as described herein can be administered to a subject by any suitable means, such as oral, intravenous, parenteral, subcutaneous, intrapulmonary, topical, intravitreal, dermal, transmucosal, rectal, and intranasal administration. Parenteral infusions include intramuscular, intravenous, intraarterial, or intraperitoneal administration. The compounds can also be administered transdermally, for example in the form of a slow-release subcutaneous implant or as a transdermal patch. They can also be administered by inhalation. Although direct oral administration may cause some loss of desired activity, the compounds can be packaged in such a way to protect the active ingredient(s) from digestion by use of enteric coatings, capsules or other methods known in the art.

Controlled-release pharmaceutical products have a common goal of improving drug therapy over that achieved by their non-controlled counterparts. The use of an optimally designed controlled-release preparation in medical treatment is characterized by a minimum of drug substance being employed to cure or control the condition in a minimum amount of time. Advantages of controlled-release formulations include extended activity of the drug, reduced dosage frequency, and increased patient compliance. In addition, controlled-release formulations can be used to affect the time of onset of action or other characteristics, such as blood levels of the drug, and can thus affect the occurrence of side (e.g., adverse) effects.

Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include, for example, the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; anti-inflammatory agents; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediamine tetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide.

The dosage can vary depending upon known factors such as the pharmacodynamic characteristics of the active ingredient and its mode and route of administration; time of administration of active ingredient; age, sex, health and weight of the recipient; nature and extent of symptoms; kind of concurrent treatment, frequency of treatment and the effect desired; and rate of excretion.

A therapeutically effective dose can depend upon a number of factors known to those of ordinary skill in the art. The dose(s) can vary, for example, depending upon the identity, size, and condition of the subject or sample being treated, further depending upon the route by which the composition is to be administered, if applicable, and the effect which the practitioner desires. These amounts can be readily determined by the skilled artisan.

In some embodiments, the therapeutically effective amount is at least about 0.1 mg/kg body weight, at least about 0.25 mg/kg body weight, at least about 0.5 mg/kg body weight, at least about 0.75 mg/kg body weight, at least about 1 mg/kg body weight, at least about 2 mg/kg body weight, at least about 3 mg/kg body weight, at least about 4 mg/kg body weight, at least about 5 mg/kg body weight, at least about 6 mg/kg body weight, at least about 7 mg/kg body weight, at least about 8 mg/kg body weight, at least about 9 mg/kg body weight, at least about 10 mg/kg body weight, at least about 15 mg/kg body weight, at least about 20 mg/kg body weight, at least about 25 mg/kg body weight, at least about 30 mg/kg body weight, at least about 40 mg/kg body weight, at least about 50 mg/kg body weight, at least about 75 mg/kg body weight, at least about 100 mg/kg body weight, at least about 200 mg/kg body weight, at least about 250 mg/kg body weight, at least about 300 mg/kg body weight, at least about 3500 mg/kg body weight, at least about 400 mg/kg body weight, at least about 450 mg/kg body weight, at least about 500 mg/kg body weight, at least about 550 mg/kg body weight, at least about 600 mg/kg body weight, at least about 650 mg/kg body weight, at least about 700 mg/kg body weight, at least about 750 mg/kg body weight, at least about 800 mg/kg body weight, at least about 900 mg/kg body weight, or at least about 1000 mg/kg body weight.

A therapeutically effective dose can depend upon a number of factors known to those of ordinary skill in the art. The dose(s) can vary, for example, depending upon the identity, size, and condition of the subject or sample being treated, further depending upon the route by which the composition is to be administered, if applicable, and the effect which the practitioner desires. These amounts can be readily determined by the skilled artisan.

A compound as described herein or composition comprising the same can be administered to the subject one time (e.g., as a single injection or deposition). Alternatively, administration can be once or twice daily to a subject in need thereof for a period of from about 2 to about 28 days, or from about 7 to about 10 days, or from about 7 to about 15 days. It can also be administered once or twice daily to a subject for a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 times per year, or a combination thereof.

Single unit dosage forms of the disclosure are suitable for oral, mucosal (e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g., subcutaneous, intravenous, bolus injection, intramuscular, or intraarterial), topical (e.g., eye drops or other ophthalmic preparations), transdermal (e.g., cream, lotion, or dermal spray) or transcutaneous administration to a subject. Examples of dosage forms include, but are not limited to: tablets; caplets; capsules, such as soft elastic gelatin capsules; cachets; troches; lozenges; dispersions; suppositories; powders; aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage forms suitable for oral or mucosal administration to a patient, including suspensions (e.g., aqueous or non-aqueous liquid suspensions or solutions, oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions, and elixirs; liquid dosage forms suitable for parenteral administration to a patient; eye drops or other ophthalmic preparations suitable for topical administration; and sterile solids (e.g., crystalline or amorphous solids) that can be reconstituted to provide liquid dosage forms for parenteral administration to a subject.

The composition, shape, and type of dosage forms of the disclosure will typically vary depending on their use. Further, the dosage can vary depending upon known factors such as the pharmacodynamic characteristics of the active ingredient and its mode and route of administration; time of administration of active ingredient; age, sex, health and weight of the recipient; nature and extent of symptoms; kind of concurrent treatment, frequency of treatment and the effect desired; and rate of excretion.

For example, a dosage form used in the acute treatment of a disease can contain larger amounts of one or more of the active agents it comprises than a dosage form used in the chronic treatment of the same disease. Similarly, a parenteral dosage form can contain smaller amounts of one or more of the active agents it comprises than an oral dosage form used to treat the same disease. These and other ways in which specific dosage forms encompassed by this disclosure will vary from one another will be readily apparent to those skilled in the art. See, e.g., Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing, Easton Pa. (1990).

Any of the therapeutic applications described herein can be applied to any subject in need of such therapy, including, for example, a mammal such as a mouse, a rat, a dog, a cat, a cow, a horse, a rabbit, a monkey, a pig, a sheep, a goat, or a human. In some embodiments, the subject is a mouse, rat, pig, or human. In some embodiments, the subject is a mouse. In some embodiments, the subject is a rat. In some embodiments, the subject is a pig. In some embodiments, the subject is a human.

Medical Kits

Aspects of the invention are directed towards a medical kit suitable for the treatment of cancer or an inflammatory condition comprising printed instructions for administering a compound as described herein to a subject in need thereof; and/or a pharmaceutical composition comprising a compound as described herein and a pharmaceutically acceptable carrier. A“kit” or “medical kit” of the disclosure comprises a dosage form of a compound of the disclosure or a pharmaceutically acceptable salt, solvate, hydrate, stereoisomer, prodrug, or clathrate thereof. A kit can also include two or more compounds as described herein, either in combination, such as in a single tablet, or provided separately, such as in two or more tablets.

Kits can further comprise additional active agents, examples of which are described herein. Kits of the disclosure can further comprise devices that are used to administer the active ingredients. Examples of such devices include, but are not limited to, syringes, drip bags, patches, and inhalers. Kits can also comprise printed instructions for administering the compound to a subject.

Kits of the invention can further comprise pharmaceutically acceptable vehicles that can be used to administer one or more active ingredients. For example, if an active ingredient is provided in a solid form that must be reconstituted for parenteral administration, the kit can comprise a sealed container of a suitable vehicle in which the active ingredient can be dissolved to form a particulate-free sterile solution that is suitable for parenteral administration. Examples of pharmaceutically acceptable vehicles include, but are not limited to: Water for Injection USP; aqueous vehicles such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

EXAMPLES

Benzil (1 Mol Eq.) was dissolved in glacial acetic acid and added to a solution of thiosemicarbazide (2.2 Mol. Eq.) in hot water. The mixture was refluxed for 4 hr., and the precipitate that appeared was filtered while hot and washed with water until GAA removed. Then it was washed with hexane to remove water. Then yellowish color solid product was obtained. The yellowish crystals obtained were recrystallized from ethanol to give orange crystals.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific substances and procedures described herein. Such equivalents are considered to be within the scope of this invention, and are covered by the following claims.