WO2003068157A2 - Kinase inhibitors and methods of use thereof - Google Patents

Abstract

The invention relates in some aspects to kinase inhibitors and methods of use thereof. The invention also relates to pharmaceutical preparations and their uses as anti-inflammatory and anti-cancer drugs.

Description

Kinase Inhibitors and Methods of Use Thereof

Related Applications

This application claims priority to U.S. Provisional Application No. 60/355,855, filed February 11, 2002, hereby incorporated by reference.

Field of the Invention

The present invention relates to kinase inhibitors and methods of use thereof. In some aspects, the invention relates to pharmaceutical preparations and their uses as anti- inflammatory and anti-cancer drugs.

Background of the Invention

Cytokines transmit signals between cells that govern aspects of their behavior. The proper transmission of these signals among immune system cells can make a critical difference in the outcomes of immune responses while inappropriate signals may cause pathologic states, such as autoimmune diseases, immunodeficiencies, and cancers. Signal transduction, the process by which cells respond when they encounter cytokines includes phosphorylation and activation of certain cellular protein kinases. One class of kinases, the Janus kinases (JAK), transduces signals from a number of cytokines including interleukins and interferons and are important for the lymphoid and myeloid cell lineages. JAK kinases play a central role in the regulation of the behavior of cells that participate in the immune response and inflammation.

Summary of the Invention

The present invention relates to kinase inhibitors, pharmaceutical preparations comprising these inhibitors, and their use as anti-inflammatory and anti-cancer drugs. In one aspect, the invention provides pharmaceutical preparations comprising a compound of Formula I:

wherein R¹ and R⁵ are H, OC(O)R\ OC(O)OR', OC(S)R', OC(S)OR', and OP(OR')(OR'); R² and R⁴ are independently selected from H, OH, SH, halo, nitro, alkyl, alkenyl, alkynyl, aryl, alkylthio, alkoxy, aryloxy, amino, NR'₂, CH₂S(CH₂)_nAr; or two of R¹, R², R³, R⁴ and R⁵ taken together are alkyl, forming a ring; R³ is selected from H, OH, SH, halo, nitro, alkyl, alkenyl, alkynyl, alkylthio, alkoxy, aryloxy, amino, NR'₂, CH₂S(CH₂)_nAr, CO₂R', COR', O(CH₂)_mCO₂R', SO₃R', OPO₃R'₂; R⁶ is selected from H, OH, SH, halo, nitro, alkyl, alkenyl, alkynyl, aryl, alkylthio, alkoxy, aryloxy, amino, NR'₂, CH₂S(CH₂)_nAr;

R⁷ and R⁸ are independently selected from H, alkyl, alkenyl, alkynyl, aryl, alkylthio, alkoxy, aryloxy, amino, NR'_2;

R⁹, R¹⁰, R¹¹, R¹², and R¹³ are independently selected from H, OH, alkyl, haloalkyl, alkenyl, alkynyl, aryl, alkylthio, alkoxy, aryloxy, amino, NR'₂, CO₂R', COR', CN, and NO₂ provided that at least one of R⁹, R¹⁰, R^u, R¹², and R¹³ is haloalkyl; or two of R⁹, R¹⁰, R¹¹, R¹², and R¹³ taken together are methylenedioxy; wherein R' is selected from H, Li⁺, Na⁺, K⁺, Mg⁺², Ca⁺², alkyl, alkenyl, alkynyl, and aryl;

' n is an integer ranging from 0 to 4; and is an integer ranging from 1 to 4; and a pharmaceutically acceptable carrier.

Although Formula I is drawn with an (E) configuration about the double bond, the formula also encompasses compounds having a (Z) orientation about the double bond.

Ar denotes an aryl group, including phenyl, benzyl, pyridyl, thienyl, furyl, indolyl, pyrrolyl, thiazolyl, and imidazolyl.

Haloalkyl encompasses Cι-C₆, branched or straight chain alkyl groups that have at least one halo substituent. Halo substituents, as used herein, encompass the atoms F, CI, Br, At, and I. Examples of the haloalkyl substiUients include CX₃, CHX₂, CH₂X, CXYZ, CHXY, CH₂CXYZ, CXYCH3 wherein X, Y, and Z are each selected from halo atoms.

In some embodiments, the haloalkyl substituents are tetrahalogenated. In other embodiments, the haloalkyl substituents are trihalogenated. In still other embodiments, the haloalkyl substituents are dihalogenated or monohalogenated.

In some embodiments, the pharmaceutical preparations of the invention comprise a compound of Formula I wherein R¹⁰ is a haloalkyl substituent. In other embodiments, R¹¹ is a haloalkyl substituent. The haloalkyl substituents, in some embodiments, are fluoroalkyl groups. In yet other embodiments, the haloalkyl substituents are CF₃. In some embodiments of the invention, pharmaceutical compositions having a compound of Formula I with hydrogen substituents at R¹ and R⁵ are provided. In some of these embodiments, the substituents at positions R⁹, R¹¹, R¹², and R¹³ are each hydrogen. In others of these embodiments, the substituents at positions R⁹, R¹⁰, R¹², and R¹³ are each hydrogen. In all of these embodiments, R² and R³ are optionally each OH. In other embodiments of the invention, R⁶, R⁷, and R⁸ are each H. In some of these embodiments R² and R³ are each OH. In some of these embodiments, R¹, R⁴, and R⁵ are each H. In still others of these embodiments, R⁹ and R¹³ are each H.

In other embodiments of the invention, R¹, R³, and R⁵ are each H. In some of these embodiments, R² and R⁴ are each OH. In some embodiments of the invention, R⁹, R¹⁰, R¹¹, R¹² are independently selected from alkoxy and hydrogen. In some of these embodiments R⁹, R¹⁰, and Rⁿ are hydrogen and R¹² and R¹³ are alkoxy.

In some embodiments of the invention, pharmaceutical preparations comprising a compound of Formula I wherein R⁷ is alkyl and R⁸ is H are provided. Generally, when R⁷ and R⁸ are different substituents, the compounds of Formula I may be a racemic mixture, or chiral compounds with an (R) configuration or an (S) configuration.

In another aspect of the invention, pharmaceutical preparations comprising a compound of Formula I is provided wherein the carbocyclic ring having the haloalkyl substituent is replaced with a heterocyclic ring having a haloalkyl substituent. Heterocyclic rings include, but are not limited to, pyridyl, thienyl, furyl, indolyl, pyrrolyl, thiazolyl, and imidazolyl. In one aspect of the invention, pharmaceutical preparations comprising a compound of Formula II:

and a pharmaceutical ly-acceptable carrier are provided.

In another aspect of the invention, a pharmaceutical preparation comprising a compound of Formula III:

and a pharmaceutically acceptable carrier are provided.

In another aspect of the invention, a pharmaceutical preparation comprising a compound of Formula V:

and a pharmaceutically acceptable carrier are provided.

According to another aspect of the invention, pharmaceutical preparations comprising compounds of Formula IV:

and a pharmaceutically-acceptable carrier are provided wherein X and Y are each independently selected from H, CH₃, CH(CH₃)₂, OCH3, CI, and F, are provided. Additionally, X and Y together may form a six-membered ring fused with the given carbocylic ring such that a naphthalene structure is formed,. Some embodiments include: (a) Y is H and X is methyl, (b) Y is methyl and X is H, (c) Y is isopropyl and X is H, (d) Y is F and X is H, (e) Y is CI and X is H, (f) Y is H and X is F, (g) Y is H and X is H, (h) Y is OCH₃ and X is H, and (i) Y is OCH₃ and X is methyl.

In one aspect of the invention compounds of Formula I, described above, are provided. In another aspect of the invention compounds of Formula IV, described above, are provided.

According to another aspect of the invention, a compound of Formula II:

is provided. In still another aspect of the invention, a compound of Formula III:

is provided.

According to yet another aspect of the invention, a compound of Formula V:

is provided. The compounds of Formulas I, II, III, IV, and V and pharmaceutical preparations comprising compounds I, II, III, IV, and V have a variety of pharmaceutical uses, including uses as therapeutic agents for a variety of conditions and disease types.

According to another aspect of the invention, methods of treating inflammatory conditions are provided. The methods involve administering to a subject in need of such treatment a pharmaceutical preparation comprising a compound of Formula I and a pharmaceutically acceptable carrier in an effective amount to treat the inflammatory condition. In one embodiment, a method of treating inflammatory conditions comprising administering to a subject in need of such treatment a pharmaceutical preparation comprising a compound of Formula II in an effective amount to treat the inflammatory condition is provided. In another embodiment, a method of treating inflammatory condition comprising administering to a subject in need of such treatment a pharmaceutical preparation comprising a compound of Formula III in an effective amount to treat the inflammatory condition is provided. In another embodiment, a method of treating inflammatory conditions comprising administering to a subject in need of such treatment a pharmaceutical preparation comprising a compound of Formula IV in an effective amount to treat the inflammatory condition is provided. In still another embodiment, a method of treating inflammatory conditions comprising administering to a subject in need of such treatment a pharmaceutical preparation comprising a compound of Formula V in an effective amount to treat the inflammatory condition is provided. Inflammatory conditions include, but are not limited to, autoimmune disorders, psoriasis, rheumatoid arthritis, experimental autoimmune encephalomyelitis (EAE), Crohn's disease, ulcerative colitis, allergic inflammatory diseases, such as asthma, excema, contact dermatitis, latex dermatitis, inflammatory bowel disease, anathylaxis, allergic rhinitis (hayfever), atopic dermatitis, graft versus host disease, and multiple sclerosis. According to another aspect of the invention, methods for treating unwanted mammalian cell proliferation are provided.

Mammalian cell proliferative disorders include cancer, psoriasis, actinic keratosis, post-angioplasty cell proliferation, and other situations involving unwanted mammalian cell proliferation as will be recognized by those of ordinary skill in the art.

In a particular embodiment, methods to treat cancer involve administering to a subject in need of such treatment a pharmaceutical preparation comprising a compound of Formula I in an effective amount to treat the cancer are provided. In one embodiment, the method of treating cancer comprises administering a compound of Formula II in an effective amount to treat cancer. In another embodiment, the method comprises administering to a subject a compound of Formula III in an effective amount to treat cancer. In still another embodiment, the method comprises administering to a subject a compound of Formula IV in an effective amount to treat cancer. In yet another embodiment, the method comprises administering to a subject a compound of Formula V in an effective amount to treat cancer. In some embodiments, the cancer is a leukemia, lymphoma, or solid tumor. Other cancers treatable by the methods of the invention include, biliary tract cancer, brain cancer, breast cancer, cervical cancer, choriocarcinoma, colon cancer, endometrial cancer, esophageal cancer, gastric cancer, hematological neoplasms, intraepithelial neoplasms, liver cancer, lung cancer, neuroblastomas, oral cancer, ovarian cancer, pancreas cancer, prostate cancer, rectal cancer, sarcomas, skin cancer, testicular cancer, thyroid cancer, and renal cancer.

As used herein, a "subject" is a human, non-human primate, dog, cat, horse, rabbit, sheep, goat, cow, pig, poultry, or rodent. In an important embodiment, the subject is a human. The compounds and pharmaceutical preparations of the invention can also be used to inhibit enzymes, such as kinases. In some embodiments, the pharmaceutical compositions and compounds of the invention are useful to inhibit kinases, including, but not limited to, JAK kinases. In some embodiments, the compounds and pharmaceutical preparations of the invention are useful to preferentially inhibit specific classes of kinases, specific kinases, or even specific JAK kinases. One of skill in the art can readily determine which compounds can preferentially inhibit specific kinases, such as JAK1, JAK2, and TYK2. The pharmaceutical preparations and compounds of the invention may optionally be used in combination with other kinase inhibitors, other nonkinase inhibitor anti-inflammatory agents or other nonkinase inhibitor anti-cancer agents.

These and other aspects of the invention will be described in more detail below.

Detailed Description

From the foregoing discussion, those of ordinary skill in the art will readily recognize that the pharmaceutical preparations, methods, and compounds of the invention have a variety of uses. The compounds can be used in vivo, and in vitro, such as the use of the compounds as laboratory tools. The compounds and preparations of the invention can be used medically or experimentally to inhibit kinases, or to inhibit to signal transduction of IL- 2, IL-3, IL-7, IL-9, IL-12, IL-15, IL-21, GM-CSF, and other cytokines.

The preparations and compounds of the invention may be used to treat disorders. In such settings, the preparations and compounds of the invention are administered to subjects in need of such treatment in effective amounts to treat the disorders. An "effective amount" as used herein means that amount necessary to delay the onset of, inhibit the progression of, halt altogether the onset or progression of, or diagnose a particular condition being treated. When administered to a subject, effective amounts will depend on the particular condition being treated and the desired outcome. It will also depend on the particular condition being treated, the severity of the condition, individual patient parameters, including age, physical condition, size, weight, concurrent treatment, frequency of treatment, mode of administration, and other factors well-known to medical practitioners. For therapeutic applications, an effective amount is that amount sufficient to achieve a medically desirable result. These factors are well-known to those of ordinary skill in the art and can be addressed with no more than routine experimentation. It is preferred generally that a maximum dose be used, that is, the highest safe dose according to sound medical judgment.

Generally, daily oral doses of active compounds will be from about 0.01 milligrams/kg per day to 1000 milligrams/kg per day. It is expected that oral doses in the range of 0.5 to 50 milligrams/kg, in one or several administrations per day, will yield the desired results. Dosage may be adjusted appropriately to achieve desired drug levels, local or systemic, depending upon the mode of administration. For example, it is expected that intravenous administration would be from an order to several orders of magnitude lower dose per day. In the event that the response in a subject is insufficient at such doses, even higher doses (or effective higher doses by a different, more localized delivery route) may be employed to the extent that patient tolerance permits. Multiple doses per day are contemplated to achieve appropriate systemic levels of compounds. The term "pharmaceutically acceptable carrier" as used herein means one or more compatible solid or liquid filler, diluents or encapsulating substances which are suitable for administration into a human or animal. The term "carrier" denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate the application. The components of the pharmaceutical compositions also are capable of being co-mingled with the molecules of the present invention, and with each other, in a manner such that there is no interaction which would substantially impair the desired pharmaceutical efficacy.

When administered, the preparations of the invention are applied in pharmaceutically acceptable amounts and in pharmaceutically acceptable compositions. Such preparations may routinely contain salts, buffering agents, preservatives, compatible carriers, and optionally other therapeutic ingredients. When used in medicine the salts should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts may conveniently be used to prepare pharmaceutically acceptable salts thereof and are not excluded from the scope of the invention. Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulphuric, nitric, phosphoric, maleic, acetic, salicylic, p-toluenesulfonic, tartaric, citric, methanesulfonic, formic, succinic, naphthalene-2-sulfonic, pamoic, 3-hydroxy-2- naphthalenecarboxylic, and benzene sulfonic. Also, pharmaceutically acceptable salts can be prepared as alkaline metal or alkaline earth salts, such as sodium, ammonium, magnesium, potassium or calcium salts of the carboxylic acid group.

Suitable buffering agents include: acetic acid and salts thereof (1-2% W V); citric acid and salts thereof (1-3% W/V); boric acid and salts thereof (0.5-2.5% W/V); and phosphoric acid and salts thereof (0.8-2% W/V).

Suitable preservatives include benzalkonium chloride (0.003-0.03% W/V); chlorobutanol (0.3-0.9% W/V); parabens (0.01-0.25% W/V) and thimerosal (0.004-0.02% W/V). A variety of administration routes are available. The particular mode selected will depend, of course, upon the particular combination of drugs selected, the severity of the condition or disorder being treated, or prevented, the condition of the patient, and the dosage required for therapeutic efficacy. The methods of this invention, generally speaking, may be practiced using any mode of administration that is medically acceptable, meaning any mode that produces effective levels of the active compounds without causing clinically unacceptable adverse effects. Such modes of administration include oral, rectal, topical, transdermal, sublingual, infusion, parenteral, intravenous, intramuscular, intracavity, as an aerosol, aural (e.g., via eardrops), intranasal, inhalation, or subcutaneous. Direct injection could also be employed for local delivery to the site of injury. Oral administration may be preferred for prophylactic treatment because of the convenience of the subject (patient) as well as the dosing schedule.

The compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the compounds of the invention into association with a carrier which constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and intimately bringing the compounds of the invention into association with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product. Compositions suitable for parenteral administration conveniently comprise a sterile aqueous preparation of the compounds of the invention is preferably isotonic with the blood of the recipient. This aqueous preparation may be formulated according to known methods using those suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenteral ly-acceptable dilutant or solvent, for example as a solution in 1, 3-butane diol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono or di-glycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. Carrier formulations suitable for oral, subcutaneous, intravenous, intramuscular, etc. are well known in the art. Compositions suitable for oral administration may be presented as discrete units such as capsules, cachets, tablets, syrups, elixirs or lozenges, each containing a predetermined amount of the compounds of the invention. Compositions suitable for any pulmonary delivery typically are formulated and/or are contained in a nebulizer. Other delivery systems can include time-release, delayed release or sustained release delivery systems. Such systems can avoid repeated administrations of the compounds of the invention, increasing convenience to the subject and the physician. Many types of release delivery systems are available and known to those of ordinary skill in the art. They include polymer based systems such as polylactic and polyglycolic acid, polyanhydrides and polycaprolactone, nonpolymer systems that are lipids including sterols such as cholesterol, liposomes; phoshpholipids; hydrogel release systems; silastic systems; peptide based system; implants and the like. Specific examples include, but are not limited to: (a) erosional systems in which the polysaccharide is contained in a form within a matrix, found in U.S. Patent Nos. 4,452,775, 4,675,189, and 5,736,152, and (b) diffusional systems in which an active component permeates at a controlled rate from a polymer such as described in U.S. Patent Nos. 3,854,480, 5,133,974 and 5,407,686. In addition, pump-based hard wired delivery systems can be used, some of which are adapted for implantation.

Use of a long-term sustained release implant may be particularly suitable for treatment of chronic conditions. "Long-term" release, as used herein, means that the implant is constructed and arranged to deliver therapeutic levels of the active ingredient for at least 7 days, and preferably 30-60 days. The implant may be positioned at the site of injury. Long-term sustained release implants are well-known to those of ordinary skill in the art and include some of the release systems described above. Important conditions treatable using the methods, preparations, and compounds of the invention include inflammatory conditions and unwanted mammalian cell proliferation.

One inflammatory condition treatable by the methods, preparations, and compounds of the invention is inflammation. "Inflammation" as used herein, is a localized protective response elicited by a foreign (non-self) antigen, and/or by an injury or destruction of tissue(s), which serves to destroy, dilute or sequester the foreign antigen, the injurious agent, and/or the injured tissue. Inflammation occurs when tissues are injured by viruses, bacteria, trauma, chemicals, heat, cold, or any other harmful stimuli. In such instances, the classic weapons of the immune system (T cells, B cells, macrophages) interface with cells and soluble products that are mediators of inflammatory responses (neutrophils, eosinophils, basophils, kinin and coagulation systems, and complement cascade).

A typical inflammatory response is characterized by (i) migration of leukocytes at the site of antigen (injury) localization; (ii) specific and nonspecific recognition of "foreign" and other (necrotic/injured tissue) antigens mediated by B and T lymphocytes, macrophages and the alternative complement pathway; (iii) amplification of the inflammatory response with the recruitment of specific and nonspecific effector cells by complement components, lymphokines and monokines, kinins, arachidonic acid metabolites, and mast cell/basophil products; and (iv) macrophage, neutrophil and lymphocyte participation in antigen destruction with ultimate removal of antigen particles (injured tissue) by phagocytosis. The ability of the immune system to discriminate between "self and "non-self (foreign) antigens is therefore vital to the functioning of the immune system as a specific defense against "non-self antigens. "Non-self antigens are those antigens on substances entering a subject, or exist in a subject but are detectably different or foreign from the subject's own constituents, whereas "self antigens are those which, in the healthy subject, are not detectably different or foreign from its own constituents. However, under certain conditions, including in certain disease states, an individual's immune system will identify its own constituents as "non-self," and initiate an immune response against "self-antigens," at times causing more damage or discomfort as from, for example, an invading microbe or foreign material, and often producing serious illness in a subject.

In one embodiment, the inflammation is caused by an immune response against a "self-antigen," and the subject in need of treatment according to the invention has an autoimmune disease. "Autoimmune disease" as used herein, results when a subject's immune system attacks its own organs or tissues, producing a clinical condition associated with the destruction of that tissue, as exemplified by diseases such as rheumatoid arthritis, uveitis, insulin-dependent diabetes mellitus, hemolytic anemias, rheumatic fever, Crohn's disease, Guillain-Barre syndrome, psoriasis, thyroiditis, Graves' disease, myasthenia gravis, glomerulonephritis, autoimmune hepatitis, multiple sclerosis, systemic lupus erythematosus, etc. Autoimmune disease may be caused by a genetic predisposition alone, by certain exogenous agents (e.g., viruses, bacteria, chemical agents, etc.), or both. Some forms of autoimmunity arise as the result of trauma to an area usually not exposed to lymphocytes, such as neural tissue or the lens of the eye. When the tissues in these areas become exposed to lymphocytes, their surface proteins can act as antigens and trigger the production of antibodies and cellular immune responses which then begin to destroy those tissues. Other autoimmune diseases develop after exposure of a subject to antigens which are antigenically similar to, that is cross-reactive with, the subject's own tissue. In rheumatic fever, for example, an antigen of the streptococcal bacterium, which causes rheumatic fever, is cross-reactive with parts of the human heart. The antibodies cannot differentiate between the bacterial antigens and the heart muscle antigens, consequently cells with either of those antigens can be destroyed.

Other autoimmune diseases, for example, insulin-dependent diabetes mellitus (involving the destruction of the insulin producing beta-cells of the islets of Langerhans), multiple sclerosis (involving the destruction of the conducting fibers of the nervous system) and rheumatoid arthritis (involving the destruction of the joint-lining tissue), are characterized as being the result of a mostly cell-mediated autoimmune response and appear to be due primarily to the action of T cells (See, Sinha et al., Science, 1990, 248:1380). Yet others, such as myesthenia gravis and systemic lupus erythematosus, are characterized as being the result of primarily a humoral autoimmune response. In some embodiments, the subject has rheumatoid arthritis, multiple sclerosis, or uveitis.

In further embodiments, the inflammation is caused by an immune response against "non-self-antigens" (including antigens of necrotic self-material), and the subject in need of treatment according to the invention is a transplant recipient, has atherosclerosis, has suffered a myocardial infarction and/or an ischemic stroke, has an abscess, and/or has myocarditis. This is because after cell (or organ) transplantation, or after myocardial infarction or ischemic stroke, certain antigens from the transplanted cells (organs) can stimulate the production of immune lymphocytes and/or autoantibodies, which later participate in inflammation/rejection (in the case of a transplant), or attack cardiac or brain target cells causing inflammation and aggravating the condition (Johnson et al., Sem. Nuc. Med. 1989, 19:238; Leinonen et al, Microbiol. Path.,1990, 9:67; Montalban et al., Stroke, 1991, 22:750). It is believed that necrotic cells from the heart or the brain may illicit a similar response. In some embodiments the compounds and pharmaceutical preparations of the invention are used to treat unwanted mammalian cell proliferation.

Important cancers treatable by the methods of the invention are: biliary tract cancer; brain cancer, including glioblastomas and medulloblastomas; breast cancer; cervical cancer; choriocarcinoma; colon cancer; endometrial cancer; esophogeal cancer; gastric cancer; hematological neoplasms, including acute lymphocytic and myelogeneous leukemia, multiple myeloma, AIDS associated leukemias and adult T-cell leukemia; lymphoma; intraepithelial neoplasms, including Bowen's disease and Paget's disease; liver cancer; lung cancer; lymphomas, including Hodgkin's disease and lymphocytic lymphomas; neuroblastomas; oral cancer, including squamous cell carcinoma; ovarian cancer, including those arising from epithelial cells, stromal cells, germ cells and mesenchymal cells; pancreas cancer; prostate cancer, rectal cancer; sarcomas, including leiomyosarcoma, rhabdomyosarcoma, liposarcoma, fibrosarcoma and osteosarcoma; skin cancer, including melanoma, Kaposi's sarcoma, basocellular cancer and squamous cell cancer; testicular cancer, including germinal tumors (seminoma, non-seminoma (teratomas, choriocarcinomas)), stromal tumors and germ cell tumors; thyroid cancer, including thyroid adenocarcinoma and medullar carcinoma; and renal cancer including adenocarcinoma and Wilms' tumor.

In general, an effective amount for treating cancer will be that amount necessary to inhibit mammalian cancer cell proliferation in situ. Those of ordinary skill in the art are well- schooled in the art of evaluating effective amounts of anti-cancer agents.

The compounds of the invention may be delivered with other other kinase inhibitors, other nonkinase inhibitor anti-inflammatory agents or other nonkinase inhibitor anti-cancer agents in the form of cocktails or individually, yet close enough in time to have a synergistic effect on the treatment. A cocktail is a mixture of any one of the compounds of the invention with another drug and/or supplementary potentiating agent which is not a compound of the invention. The use of such cocktails in the treatment of cancer and inflammatory conditions is routine. In this embodiment, a common administration vehicle (e.g., pill, tablet, implant, injectable solution, etc.) could contain both compound of the invention and the other anti- cancer, supplementary potentiating, or anti-inflammatory agent. ^• Anti-cancer agents include anti-cancer drugs. Anti-cancer drugs are well known and include: Acivicin; Aclarubicin; Acodazole Hydrochloride; Acronine; Adozelesin; Aldesleukin; Altretamine; Ambomycin; Ametantrone Acetate; Aminoglutethimide; Amsacrine; Anastrozole; Anthramycin; Asparaginase; Asperlin; Azacitidine; Azetepa;

Azotomycin; Batimastat; Benzodepa; Bicalutamide; Bisantrene Hydrochloride; Bisnafide

Dimesylate; Bizelesin; Bleomycin Sulfate; Brequinar Sodium; Bropirimine; Busulfan;

Cactinomycin; Calusterone; Caracemide; Carbetimer; Carboplatin; Carmustine; Carubicin Hydrochloride; Carzelesin; Cedefingol; Chlorambucil; Cirolemycin; Cisplatin; Cladribine;

Crisnatol Mesylate; Cyclophosphamide; Cytarabine; Dacarbazine; Dactinomycin;

Daunorubicin Hydrochloride; Decitabine; Dexormaplatin; Dezaguanine; Dezaguanine

Mesylate; Diaziquone; Docetaxel; Doxorubicin; Doxorubicin Hydrochloride; Droloxifene;

Droloxifene Citrate; Dromostanolone Propionate; Duazomycin; Edatrexate; Eflornithine Hydrochloride; Elsamitrucin; Enloplatin; Enpromate; Epipropidine; Epirubicin

Hydrochloride; Erbulozole; Esorubicin Hydrochloride; Estramustine; Estramustine Phosphate

Sodium; Etanidazole; Etoposide; Etoposide Phosphate; Etoprine; Fadrozole Hydrochloride;

Fazarabine; Fenretinide; Floxuridine; Fludarabine Phosphate; Fluorouracil; Flurocitabine;

Fosquidone; Fostriecin Sodium; Gemcitabine; Gemcitabine Hydrochloride; Hydroxyurea; Idarubicin Hydrochloride; Ifosfamide; Ilmofosine; Interferon Alfa-2a; Interferon Alfa-2b;

Interferon Alfa-nl ; Interferon Alfa-n3; Interferon Beta-I a; Interferon Gamma-I b; Iproplatin;

Irinotecan Hydrochloride; Lanreotide Acetate; Letrozole; Leuprolide Acetate; Liarozole

Hydrochloride; Lometrexol Sodium; Lomustine; Losoxantrone Hydrochloride; Masoprocol;

Maytansine; Mechlorethamine Hydrochloride; Megestrol Acetate; Melengestrol Acetate; Melphalan; Menogaril; Mercaptopurine; Methotrexate; Methotrexate Sodium; Metoprine;

Meturedepa; Mitindomide; Mitocarcin; Mitocromin; Mitogillin; Mitomalcin; Mitomycin;

Mitosper; Mitotane; Mitoxantrone Hydrochloride; Mycophenolic Acid; Nocodazole;

Nogalamycin; Ormaplatin; Oxisuran; Paclitaxel; Pegaspargase; Peliomycin; Pentamustine;

Peplomycin Sulfate; Perfosfamide; Pipobroman; Piposulfan; Piroxantrone Hydrochloride; Plicamycin; Plomestane; Porfimer Sodium; Porfiromycin; Prednimustine; Procarbazine

Hydrochloride; Puromycin; Puromycin Hydrochloride; Pyrazofϊirin; Riboprine; Rogletimide;

Safingol; Safingol Hydrochloride; Semustine; Simtrazene; Sparfosate Sodium; Sparsomycin;

Spirogermanium Hydrochloride; Spiromustine; Spiroplatin; Streptonigrin; Streptozocin;

Sulofenur; Talisomycin; Tecogalan Sodium; Tegafur; Teloxantrone Hydrochloride; Temoporfin; Teniposide; Teroxirone; Testolactone; Thiamiprine; Thioguanine; Thiotepa;

Tiazofurin; Tirapazamine; Topotecan Hydrochloride; Toremifene Citrate; Trestolone

Acetate; Triciribine Phosphate; Trimetrexate; Trimetrexate Glucuronate; Triptorelin; Tubulozole Hydrochloride; Uracil Mustard; Uredepa; Vapreotide; Verteporfin; Vinblastine Sulfate; Vincristine Sulfate; Vindesine; Vindesine Sulfate; Vinepidine Sulfate; Vinglycinate Sulfate; Vinleurosine Sulfate; Vinorelbine Tartrate; Vinrosidine Sulfate; Vinzolidine Sulfate; Vorozole; Zeniplatin; Zinostatin; and Zorubicin Hydrochloride. Other anti-cancer drugs include: 20-epi-l,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein- 1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorlns; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine; clomifene analogues; clotrimazole; collismycin A; collismycin B; combretastatin A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin; dexifosfamide; dexrazoxane; dexverapamil; diaziquone; didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine; dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docosanol; dolasetron; doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride; estramustine analogue; estrogen agonists; estrogen antagonists; etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride; flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane; fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine; ilomastat; imidazoacridones; imiquimod; immunostimulant peptides; insulin-like growth factor-1 receptor inhibitor; interferon agonists; interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; irinotecan; iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemia inhibiting factor; leukocyte alpha interferon; leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole; linear polyamine analogue; lipophilic disaccharide peptide; lipophilic platinum compounds; lissoclinamide 7; lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysin inhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone; meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone; mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonal antibody, human chorionic gonadotrophin; monophosphoryl lipid A+myobacterium cell wall sk; mopidamol; multiple drug resistance gene inhibitor; multiple tumor suppressor 1 -based therapy; mustard anticancer agent; mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N- substituted benzamides; nafarelin; nagrestip; naloxone +pentazocine; napavin; naphterpin; nartograstim; nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn; O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin; osaterone; oxaliplatin; oxaunomycin; paclitaxel analogues; paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A; placetin B; plasminogen activator inhibitor; platinum complex; platinum compounds; platinum-triamine complex; porfimer sodium; porfiromycin; propyl bis-acridone; prostaglandin J2; proteasome inhibitors; protein A-based immune modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists; raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide; rohitukine; romurtide; roquinimex; rubiginone Bl ; ruboxyl; safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine; senescence derived inhibitor 1 ; sense oligonucleotides; signal transduction inhibitors; signal transduction modulators; single chain antigen binding protein; sizofiran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-cell division inhibitors; stipiamide; stromelysin inhibitors; sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; synthetic glycosaminoglycans; tallimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomerase inhibitors; temoporfin; temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thalidomide; thiocoraline; thrombopoietin; thrombopoietin mimetic; thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocene dichloride; topotecan; topsentin; toremifene; totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase receptor antagonists; vapreotide; variolin B; vector system, erythrocyte gene therapy; velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer. Kinase inhibitors include, but are not limited to, inhibitors of JAK kinases, Cdc7 kinases, KSS1 kinases, ERK kinases, abl kinases, cdk2 kinases, cdc2 kinases, cyclic-GMP- dependent kinases, Ca²⁺/calmodulin-dependent kinases, myosin light chain kinases, TGF-β receptor kinases, Mos kinases, Raf kinases, Lck kinases, Src kinases, EGF receptor kinases, PDGF receptor kinases, Weel kinases, tyrosine kinases, cyclic AMP-dependent kinases, protein kinase C, adenosine kinases, as well as other kinase inhibitors. Some specific examples of kinase inhibitors include STI571 (Gleevec™), N-(trifluoromethylphenyl)-5- methylisoxazol-4-carboxamide, 3-[(2,4-dimethylpyrrol-5-yl)methylidenyl)indolin-2-one, 17- (allyIamino)-17-demethoxygeldanamycin, 4-(3-chloro-4-fluorophenylamino)-7-methoxy-6- [3-(4-morpholinyl)propoxyl]quinazoline, N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4- quinazolinamine, BIBX1382, 2,3,9,10,1 l,12-hexahydro-10-(hydroxymethyl)-10-hydroxy-9- methyl-9, 12-epoxy -1 H-diindolo[ 1 ,2,3-fg:3',2', 1 '-kl]pyrrolo[3,4-i][ 1 ,6]benzodiazocin- 1 -one, SH268, genistein, CEP2563, 4-(3-chlorophenylamino)-5,6-dimethyl-7H-pyrrolo [2,3- d]pyrimidinemethane sulfonate, 4-(3-bromo-4-hydroxyphenyl)amino-6,7- dimethoxyquinazoline, 4-(4'-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, SU6668, N-4- chlorophenyl-4-(4-pyridylmethyl)-l-phthalazinamine, EMD121974, H7, staurosporine, SP- 203580, PD98059, isoflavones such as, CGP 41251, fiavopiridol, p21, cipl, olomoucine, p27kipl, MLCK inhibitors, iodotubercidin, H7, and indolocarbazoles.

Anti-inflammatory agents include: Alclofenac; Alclometasone Dipropionate; Algestone Acetonide; Alpha Amylase; Amcinafal; Amcinafide; Amfenac Sodium; Amiprilose Hydrochloride; Anakinra; Anirolac ; Anitrazafen; Apazone; Balsalazide Disodium; Bendazac; Benoxaprofen ; Benzydamine Hydrochloride; Bromelains; Broperamole; Budesonide; Carprofen; Cicloprofen; Cintazone; Cliprofen; Clobetasol

Propionate; Clobetasone Butyrate; Clopirac; Cloticasone Propionate; Cormethasone Acetate; Cortodoxone; Deflazacort; Desonide; Desoximetasone; Dexamethasone Dipropionate; Diclofenac Potassium; Diclofenac Sodium; Diflorasone Diacetate; Diflumidone Sodium; Diflunisal ; Difluprednate; Diftalone; Dimethyl Sulfoxide; Drocinonide; Endrysone; Enlimomab ; Enolicam Sodium ; Epirizole ; Etodolac; Etofenamate ; Felbinac; Fenamole; Fenbufen; Fenclofenac; Fenclorac; Fendosal; Fenpipalone; Fentiazac; Flazalone; Fluazacort; Flufenamic Acid; Flumizole; Flunisolide Acetate; Flunixin ; Flunixin Meglumine ; Fluocortin Butyl; Fluorometholone Acetate; Fluquazone; Flurbiprofen ; Fluretofen; Fluticasone Propionate; Furaprofen; Furobufen; Halcinonide; Halobetasol Propionate; Halopredone Acetate; Ibufenac ; Ibuprofen; Ibuprofen Aluminum; Ibuprofen Piconol; Ilonidap;

Indomethacin; Indomethacin Sodium; Indoprofen ; Indoxole ; Intrazole; Isoflupredone Acetate; Isoxepac; Isoxicam; Ketoprofen; Lofemizole Hydrochloride ; Lornoxicam ; Loteprednol Etabonate; Meclofenamate Sodium; Meclofenamic Acid; Meclorisone Dibutyrate; Mefenamic Acid ; Mesalamine; Meseclazone; Methylprednisolone Suleptanate; Morniflumate; Nabumetone; Naproxen ; Naproxen Sodium ; Naproxol ; Nimazone; Olsalazine Sodium; Orgotein ; Orpanoxin; Oxaprozin; Oxyphenbutazone; Paranyline Hydrochloride; Pentosan Polysulfate Sodium; Phenbutazone Sodium Glycerate; Pirfenidone ; Piroxicam; Piroxicam Cinnamate; Piroxicam Olamine; Pirprofen; Prednazate; Prifelone; Prodolic Acid; Proquazone; Proxazole; Proxazole Citrate ; Rimexolone; Romazarit ; Salcolex ; Salnacedin; Salsalate ; Sanguinarium Chloride ; Seclazone ; Sermetacin; Sudoxicam; Sulindac; Suprofen; Talmetacin; Talniflumate ; Talosalate ; Tebufelone ; Tenidap; Tenidap Sodium; Tenoxicam; Tesicam; Tesimide; Tetrydamine ; Tiopinac ; Tixocortol Pivalate;

Tolmetin; Tolmetin Sodium; Triclonide; Triflumidate; Zidometacin; and Zomepirac Sodium.

As used herein, alkyl, alkenyl, alkynyl, alkylthio, and alkoxy refer to hydrocarbons Ci-Cβ in length, substituted or branched. Typical substituents include, but are not limited to, halo, hydroxy, as well as thio and amino groups. Aryl refers to a group, which may be substituted, comprising a benzene ring structure, such as benzyl and napthalyl, and having from 6 to 20 carbon atoms.

The starting materials for synthesizing the compounds of the invention are generally available, and most can be obtained, for example, from Aldrich Chemical Co. (St. Louis, MO) and Novabiochem (San Diego, CA). The compounds of Formulas I, II, III, IV, and V can be prepared following the procedures of Shi et al., Biotechnology and Bioengineering (Combinatorial Chemistry), 1998, 61(1):7-12. As will be recognized by those of skill in the art, the compounds may be synthesized, isolated and/or purified using any known method using no more than routine skill. Chiral compounds may be optionally enantiomerically enhanced or purified as well.

Example 1

Identification of Kinase Inhibitory Activity A combinatorial chemical library of six hundred variants of the known JAK kinase inhibitor AG490 was produced following the directed sorting procedure of Shi et al. (supra). 1.5 μmole of each compound was placed in a 96-well microtiter plate and dissolved in dimethylsulfoxide.

The ability of each compound to inhibit JAK2 and JAK3 kinases was determined using a cell-based assay with two factor-dependent urine cell lines, Baf3 (Palacios, R., et al., Interleukin-3 supports growth of mouse pre-B-cell clones in vitro. Nature, 1984. 309:126- 31. and Palacios, R. and M. Steinmetz, Il-3-dependent mouse clones that express B-220 surface antigen, contain Ig genes in germ-line configuration, and generate B lymphocytes in vivo. Cell, 1985. 4J.:727-34.) and 2E8 (Pietrangeli, C.E., et al, Stromal cell lines which support lymphocyte growth: characterization, sensitivity to radiation and responsiveness to growth factors. Eur J Immunol, 1988.18:863-72). The Baf3 cell line is dependent upon IL-3, which utilizes the JAK2 kinase. The 2E8 cell line is dependent upon IL-7, which utilizes the JAK3 kinase. The factor-dependent cells were maintained in Dulbecco's Modified Eagle medium with 10% fetal calf serum and saturating levels of recombinant murine IL-7 or IL-3, as appropriate.

Assays were performed by incubating 10,000 to 100,000 cells in 0.1 ml of media in the presence or absence of IL-7 or IL-3 and the test compounds. After 40 hours, 0.5 μCi of ³H-thymidine (New England Nuclear, NET 027, 6.7 Ci/mmol) in 25 μl of media was added to each well. After 8 additional hours, the cultures were harvested, and the cells were lysed by hypotonic shock. The macromolecules were captured on glass fiber filters using a Tomtec automated harvester. The radioactivity incorporated was determined by scintillation counting using a Wallac filtermat counter. Figure 1 shows the ³H-thymidine uptake of IL-7- dependent 2E8 cells in the presence of AG490 (D), Formula II (o) and Formula III (0). Incorporated ³H-thymidine was quantitated by scintillation counting and is plotted on the y- axis in Figure 1. Formulas II and III were found to be approximately 4 to 8 times more potent inhibitors of both cell lines than the known kinase inhibitor, AG490. The IC50S measured for Formulas I and II are each less than 5 μM.

The IC₅₀ measured for this preparation of AG490 was approximately 20 μM. The AG490 used in this experiment was prepared as a component of the combinatorial library. Other experiments using commercial AG490 (Sigma) have yielded an IC50 of 10 μM. The discrepancy in the measurement is most likely because the combinatorial preparations were not fully purified.

The relative inhibition measured for several compounds is shown in Table 1 and exemplary structures and their percent inhibition are listed directly below Table 1 :

Table 1

490 43% Inhibition

B9 100% Inhibition

5H4 100% Inhibition

5A1 1 98% Inliibition

7C9 95% Inhibition

2D7 92% Inhibition

10

6E1183% Inhibition

15

Example 2

Identification of Preferential Kinase Inhibitory Activity Four JAK kinases, JAK1, JAK2, JAK3, and TYK2, are intracellular molecules that regulate the behavior of numerous types of cells in response to specific extracellular signals. These kinases are thought to mediate the pathological behavior of lymphoid and myeloid cells. JAK3 kinase is unique in that it has been found to be normally expressed only in lymphoid cells. Inflammatory responses and malignancies involving lymphoid cells are often dependent upon the action of JAK3 kinase. Accordingly, inhibitors of JAK3 kinase are thought to be valuable therapeutic agents. Most inhibitors of JAK3 kinase also inhibit JAK2 kinase. Since JAK2 kinase is thought to be required for the action of certain vital signaling molecules, such as erythropoeitin, it is desirable to have selective inhibitors of JAK3 that do not inhibit JAK2. Selective inhibitors of JAK3 that do not interfere with JAK2 may be useful as therapeutic agents in the treatment of, for example, inflammatory disorders and lymphoid malignancies.

While JAK2 is an essential component of the signal transduction apparatus by which many different types of cells respond to several different cytokines, JAK3 expression is only observed in lymphoid cells. Therefore a JAK3-specific inhibitor would only affect (or preferentially affect) lymphocytes and not interfere as much or at all with the action of JAK2 in various non-lymphoid cells. The different roles of JAK2 and JAK3 are highlighted by the phenotypes of mice lacking functional JAK2 or JAK3 genes. JAK2 is required for erythropoietin signaling and accordingly JAK2 deficient mouse die from anemis as embryos (Neubauer et al, Cell, 93(3): 397-409 (1998)). In contrast, JAK3 deficient mice have a severe deficiency in lymphocytes but are otherwise viable (Thomis et al, Science, 270 (5237):794-7 (1995)).

While not wishing to be bound by theory, a selective inhibitor of JAK3 is thought to be capable of suppressing the cellular responses of lymphocytes to IL-2, IL-4, IL-7, IL-9, 1L- 15, and IL21, thereby inhibiting the growth and action of normal, pathologic or malignant lymphocytes while avoid toxic side effects of inhibition of JAK2. The library of AG490 variants, described above, was screened for inhibition of proliferation of IL-7-dependent 2E8 cells (TIB-230, ATCC, Manassis, VA) and IL-3- dependent Ba/F3 cells (ACC 300, DSMZ, Braunschweig, Germany). A unique derivative of AG490 was identified, 5F5 (Formula V), that has diminished activity as an inhibitor of IL-3 - dependent proliferation of Ba/F3 cells yet retains its activity as an inhibitor of IL-7-dependent proliferation of 2E8 cells. 2E8 cells and Ba/F3 cells were grown in the presence of AG490 or compound 5F5 (Formula V). Cells were cultured in the presence of excess concentrations of recombinant murine IL-3 (Ba/F3) or IL-7 (2E8) and varying concentrations of tyrphostins AG490 or 5F5. Cell proliferation was measured as ³H-thymidine uptake. The responses of both cell lines to increasing concentrations of AG490 are indistinguishable. The response of the 2E8 cells to 5F5 is comparable to their response to AG490, however, the Ba/F3 cells are affected significantly less by 5F5 than they are by AG490. These results indicate that 5F5 is a more potent inhibitor of JAK3 than it is of JAK2. 5F5 has been shown to be more than 10 times more effective at inhibiting JAK3 than it is an inhibiting JAK2.

Each of the patents, patent applications and references that are recited in this application are herein incorporated in their entirety by reference. Having described the presently preferred embodiments, and in accordance with the present invention, it is believed that other modifications, variations and changes will be suggested to those skilled in the art in view of the teachings set forth herein. It is, therefore, to be understood that all such variations, modifications, and changes are believed to fall within the scope of the present invention as defined by the appended claims. What is claimed is:

Claims

1. A pharmaceutical preparation comprising a compound of the formula:

wherein R¹ and R⁵ are H; R² and R⁴ are independently selected from H, OH, SH, halo, nitro, alkyl, alkenyl, alkynyl, aryl, alkylthio, alkoxy, aryloxy, amino, NR'₂, CH₂S(CH₂)_nAr; or two of R¹, R², R³, R⁴ and R⁵ taken together are alkyl, forming a ring; R³ is selected from H, OH, SH, halo, nitro, alkyl, alkenyl, alkynyl, alkylthio, alkoxy, aryloxy, amino, NR'₂, CH₂S(CH₂)_nAr, CO₂R', COR', O(CH₂)_mCO₂R', SO₃R\ OPO₃R'₂; R⁶ is selected from H, OH, SH, halo, nitro, alkyl, alkenyl, alkynyl, aryl, alkylthio, alkoxy, aryloxy, amino, NR'₂, CH₂S(CH₂)_nAr;

R⁷ and R⁸ are independently selected from H, alkyl, alkenyl, alkynyl, aryl, alkylthio, alkoxy, aryloxy, amino, NR'₂;

R⁹, R¹⁰, R¹¹, R¹², and R¹³ are independently selected from H, OH, alkyl, haloalkyl, alkenyl, alkynyl, aryl, alkylthio, alkoxy, aryloxy, amino, NR'₂, , CO₂R', COR', CN, and NO₂ provided that at least one of R⁹, R¹⁰, R¹¹, R¹², and R¹³ is haloalkyl; or two of R⁹, R¹⁰, R^π, R¹², and R¹³ taken together are methylenedioxy;

R' is selected from H, Li⁺, Na⁺, K⁺, Mg⁺², Ca⁺², alkyl, alkenyl, alkynyl, and aryl; n is an integer ranging from 0 to 4; and m is an integer ranging from 1 to 4; and a pharmaceutically acceptable carrier.

2. A pharmaceutical preparation according to claim 1 wherein the haloalkyl has a formula selected from CX₃, CHX₂, CH₂X, CXYZ, CHXY, CH₂CXYZ, CXYCH₃ wherein X, Y, and Z are independently selected from F, Cl, Br and I.

3. A pharmaceutical preparation according to claim 1 wherein the haloalkyl is tetrahalogenated.

4. A pharmaceutical preparation according to claim 1 wherein the haloalkyl is trihalogenated.

5. A pharmaceutical preparation according to claim 1 wherein the haloalkyl is dihalogenated.

6. A pharmaceutical preparation according to claim 1 wherein the haloalkyl is monohalogenated.

7. A pharmaceutical preparation according to claim 1 wherein R¹⁰ is haloalkyl.

8. A pharmaceutical preparation according to claim 1 wherein R¹ ' is haloalkyl.

9. A pharmaceutical preparation according to claim 7 or 8 wherein the haloalkyl is a fluoroalkyl.

10. A pharmaceutical preparation according to claim 9 wherein the haloalkyl is CF₃.

Q 1 1 1 11 11. A pharmaceutical preparation according to claim 7 wherein the R , R , R , and R are each H.

12. A pharmaceutical preparation according to claim 8 wherein the R⁹, R¹⁰, R¹², and R¹³ are each H.

13. A pharmaceutical preparation according to claim 1 wherein R² and R³ are each OH.

14. A pharmaceutical preparation according to claim 1 or 13 wherein R¹, R⁴ and R⁵ are each H.

15. A pharmaceutical preparation according to claim 1 or 13 wherein R⁶, R⁷, and R⁸ are each H.

16. A pharmaceutical preparation according to claim 15 wherein R and R are each H.

17. A pharmaceutical preparation according to claim 1 or 13 wherein R⁷ is alkyl and R⁸ is H.

18. A pharmaceutical preparation according to claim 17 wherein the compound has a stereocenter with an (R) configuration.

19. A pharmaceutical preparation according to claim 17 wherein the compound has a stereocenter with an (S) configuration.

20. A pharmaceutical preparation according to claim 1 comprising a compound of the formula:

and a pharmaceutically acceptable carrier.

21. A pharmaceutical preparation according to claim 1 comprising a compound of the formula:

and a pharmaceutically acceptable carrier.

22. A pharmaceutical preparation according to claim 1 comprising a compound of the formula:

and a pharmaceutically acceptable carrier.

23. A method of treating an inflammatory condition comprising administering to a subject in need of such treatment a pharmaceutical preparation of any one of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and 22 in an effective amount to treat the inflammatory condition.

24. A method of treating an inflammatory condition comprising administering to a subject in need of such treatment a compound of the formula:

in an effective amount to treat the inflammatory condition.

25. A method of treating an inflammatory condition comprising administering to a subject in need of such treatment a compound of the formula:

in an effective amount to treat the inflammatory condition.

26. A method of treating an inflammatory condition comprising administering to a subject in need of such treatment a compound of the formula:

in an effective amount to treat the inflammatory condition.

27. A method of treating unwanted mammalian cell proliferation comprising administering to a subject in need of such treatment a pharmaceutical preparation of any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and 22 in an effective amount to treat the cell proliferation.

28. The method of claim 27 wherein the unwanted mammalian cell proliferation is a cancer.

29. The method of claim 28 wherein the cancer is a leukemia, a lymphoma, or a solid tumor.

30. A method of treating unwanted mammalian cell proliferation comprising administering to a subject in need of such treatment a compound of the formula:

in an effective amount to treat the cell proliferation.

31. A method of treating unwanted mammalian cell proliferation comprising administering to a subject in need of such treatment a compound of the formula:

in an effective amount to treat the cell proliferation.

32. A method of treating unwanted mammalian cell proliferation comprising administering to a subject in need of such treatment a compound of the formula:

in an effective amount to treat the cell proliferation.

33. A method of any one of claims 30, 31, and 32 wherein the unwanted mammalian cell proliferation is a cancer.

34. The method of claim 33, wherein the cancer is a leukemia, a lymphoma, or a solid tumor.

35. A compound of the formula:

36. A compound of the formula:

37. A compound of the formula: