WO2020123277A1

WO2020123277A1 - Protein kinase inhibitor prodrugs

Info

Publication number: WO2020123277A1
Application number: PCT/US2019/064866
Authority: WO
Inventors: Kan He
Original assignee: Princeton Drug Discovery Inc.
Priority date: 2018-12-10
Filing date: 2019-12-06
Publication date: 2020-06-18

Abstract

The present invention relates to ester and phosphate prodrugs of the protein kinase inhibitor N-(2-chloro-6-methylphenyl)-2-[[2-methyl-6-((S)-3-hydroxypyrrolidin-1- yl)pyrimidin4-yl]amino]thiazole-5-carboxarnide (compound X), stereoisomers, isomeric mixtures including racemic mixtures, and/or 4-deutero substituted derivatives thereof.

Description

PROTEIN KINASE INHIBITOR PRODRUGS

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority from U.S. Provisional application number

62/777,385 filed on December 10, 2018, the entire contents of which is incorporated herein by reference.

FIELD OF THE INVENTION

[0001] The present invention relates to prodrugs of the protein kinase inhibitor N (2-chloro-6-methylpheny!)-2-H2-mefoyl~6-((S)-3-hydroxypyrro!idin-1-yl)pyrimidin- 4-yl]amino]tiiiazoie-5-carboxamrde (compound X), stereoisomers, isomeric mixtures including racemic mixtures, and/or 4-deutero substituted derivatives thereof.

BACKGROUND OF THE INVENTION

[0002] The information provided herein is intended solely to assist the

understanding of the reader. None of the information provided nor references cited is admitted to be prior art to the present invention.

[0003] The identification of the molecular events that underlie the development of human diseases presente a major challenge in the design of improved strategies in the prevention, management, and cure of certain diseases (Lahiry P et al. Kinase mutations in human disease: interpreting genotype-phenotype

relationships. Nat Rev Genet 2010; H(t):60-74).

[0004] The role of aberrantly regulated protein tyrosine kinases (PTKs) in human diseases is the subject of intense investigation (Lahiry id ). Protein kinases are regulators of cellular signaling, and their functional dysregulation is common in carcinogenesis, autoimmune reactions, and many other disease states or conditions (Lahiry id.; Vargas L et al. Inhibitors of BTK and ITK: state of the new drugs for cancer, autoimmunity and inflammatory diseases. Scand J Immunol. 2013; 78(2): 130-9; Nobel ME et al. Protein kinase inhibitors: insights into drug design from structure. Science.2004; 303:1800-1805). The human genome encodes over 500 protein kinases that share a catalytic domain conserved in sequence and structure but which are notably different in how their catalysis is regulated (Manning G et al. The protein kinase complement of the human genome. Science. 2002; 298:1912-1934; Nobel id.). Protein kinases regulate key signal transduction cascades that control or are involved in the control of physiological functions, including cellular growth and proliferation, ceil differentiation, cellular development, cell division, stress response, transcription regulation, aberrant mitogenesis, angiogenesis, abnormal endothelial cell-cell or cell-matrix interactions during vascular development, inflammation, Jun-N- terminal kinase (JNK) signal transduction, and several other cellular processes

(Manning id). Protein kinase inhibitors have been established as promising drugs that inhibit overactive protein kinases in cancer cells (Gross S et al. Targeting cancer with kinase inhibitors. J Clin Invest. 2015;125(5):1780-1789; Vargas id).

[0005] A partial, non-limiting (1st of kinases includes: ABL, ACK, ARG, BLK, BMX, BRK, BTK, GSK, DDR1, DDR2, EGFR, EPHA1, FGR, FMS, FRK, FYN(isOfbrm a), FYN(isoform b), HCK, KIT, LCK, LYNa, PDGFRa, PDGFRp, SRC, SRM,

YES» PIK3CA/PIK3R1 (Manning id). Aberrant kinase activity has been observed in many disease states including benign and malignant proliferative conditions as well as diseases resulting from inappropriate activation of the immune and nervous systems.

[0006] Prodrug design is a widely known molecular modification strategy that aims to optimize the physicochemical and pharmacological properties of drugs to improve their solubility and pharmacokinetic features and decrease their toxicity. A lack of solubility is one of the main obstacles to drug development (Jornada DH_> et al.“The Prodrug Approach: A Successful Tool for improving Drug

Solubility" Molecules 2016, vot. 21, 42; doi:10.3390/molecules21010042), A prodrug is a poorly active or inactive compound containing the parental drug that undergoes some in vivo biotransfbrmation through chemical ex enzymatic cleavage, enabling the delivery of the active molecule at efficacious levels (Jornada id). The classical approach for prodrug design uses the non-specific strategy of covalently modifying the drug of interest by attaching hydrophilic functionalities (e.g., phosphate, sulfate) to increase the solubility of the parent drug, or lipophilic moieties (e.g., ester) to increase its passive permeability (Dahan A, est al.‘Modem Prodrug Design for Targeted Oral Drug Delivery” Molecules 2014, vol. 19, 16489-16505; doi:10.3390/molecules191016489).

SUMMARY OF INVENTION

[00071 The present Invention concerns novel ester and phosphate prodrugs of the protein kinase inhibitor N-(2-chloro-6-methy}phenyl)-2-[[2-methyl-6 ((S)-3- hydroxypyrrolidin-1-yl)pyrimidin-4-yi]aminoJthiazde-5-cartK>xamide (Compound X), stereoisomers, isomeric mixtures including racemic mixtures, and/or 4- deutero substituted derivatives thereof. Compound X has the following structure:

Ester prodrugs of Compound X have the following structure:

Ester Prodrugs of Compound X where R is methyl, ethyl, propyl, or butyl.

Phosphate prodrug of Compound X has the following structure:

[0008] Compounds IV, V, X, and XI inhibit the activity of one or more protein kinases and are expected to be useful in treating kinase-related diseases or conditions. Compounds IV, V, X, and XI are disclosed in US 10,479,786 B2, US

10,174,018 B2, US 2019/0071437 A1, and US 2018/0099961 A1.

BRIEF DESCRIPTION OF THE FIGURES

[0009] FIG 1, LC-MS/MS Chromatogram and Spectra oM 475-13 Incubated with Human Hepatocytes at 0-hour (Control).

[0010] FIG 2. LC-MS/MS Chromatogram and Spectra of 1475-13 Incubated with Human Hepatocytes for 1 hour.

[0011] FIG 3. LC-MS/MS Chromatogram and Spectra of 1475-13 Incubated with Hurhan Hepatocytes for 2 hours;

[0012] FIG 4. LC-MS/MS Chromatogram and Spectra of 1475-16 Incubated with Mouse Hepatocytes for 0 and 1 hour.

[0013] FIG 5. LC-MS/MS Chromatogram and Spectra of 1475-16 Incubated with Human Hepatocytes for 0 and 1 hour.

DESCRIPTION OF THE INVENTION

Definitions [0014] As used herein the following definitions apply unless clearly indicated otherwise. By“chemical structure” or“chemical substructure" is meant any definable atom or group of atoms that constitute an individually identifiable molecule, portion of a molecule, such as a substituent moiety, a core which is optionally substituted, and the like. Normally, chemical substructures of a ligand can have a role in binding of the ligand to a target molecule, or can influence toe three-dimensional shape, electrostatic charge, and/or conformational properties of the ligand.

[0015] The term "prodrug” is a compound that, upon in vivo administration, is metabolized by one or more steps or processes or otherwise converted to the biologically, pharmaceutically, or therapeutically active form of the compound. To produce a prodrug, the pharmaceutically active compound is modified such that the active compound will be regenerated by metabolic or hydrolytic processes.

[0016] The term“binds," in connection with the interaction between a target and a potential binding compound, indicates that the potential binding compound associates with the target to a statistically significant degree as compared to association with proteins generally (i.e., non-specific binding).

[0017] As used herein, the term“modulating" or“modulate” refers to an effect of altering a biological activity, especially a biological activity associated with a particular biomolecule such as a protein kinase. For example, an agonist or antagonist of a particular biomoiecuie modulates the activity of that biomolecule, e.g., an enzyme, by either increasing (e.g., agonist, activator), or decreasing (e.g., antagonist, inhibitor) its activity. This type of activity is typically indicated in terms Of an half maximal effective concentration (EG») or half maximal inhibitory concentration (ICso) for an activator or inhibitor, respectively. Additionally, inhibition activity can be expressed in percent inhibition and/or Ki.

[0018] As used herein in connection with compounds of the invention, the term “synthesizing* and like terms means chemical synthesis from one or more precursor materials. Further, by“assaying" is meant the creation of experimental conditions and the gathering of data regarding a particular result of the

experimental conditions. For example, enzymes can be assayed based cm their ability to act upon a detectable substrate. A compound or ligand csin be assayed based on its ability to bind to a particular target molecule or molecules.

[0019]“D,”“d,” and“²H^* refer to a deuterium atom, a stable isotope of hydrogen with a mass twee that of hydrogen (atomic weight of 2.0144). Hydrogen naturally occurs as a mixture of the isotopes hydrogen (¹H), deuterium (²H or D), and tritium (³H or T). The natural abundance of deuterium is about 0.015%. A person skilled in the art would recognize that all chemical compounds with a hydrogen atom actually are present as mixtures of the H and D isotopes, with about 0.015% being the deuterium isotope. Compounds with a level of deuterium that has been enriched to be greater than its natural abundance of

0.015% should be considered unnatural, and as a result novel, over their non- enriched counterparts. The D in structural formulas and chemical compounds herein refers to incorporation of D in amounts greater than 0.015%.

[00201 It is to be understood that the compounds provided herein may contain chiral centers. Such chiral centers may be of either the (R) or (S) configuration, or may be a mixture thereof. Thus, the compounds provided herein may be enantiomerically pure, or stereoisomeric or diastereomeric mixtures thereof, including racemic mixtures (about 50:50 ratio of enantiomers).

[0021] The term“pharmaceutically acceptable” means that the indicated material does not have proparties that would cause a reasonably prudent medical practitioner to avoid administration of the material to a patient, taking into consideration the disease or condition to be treated and the respective route of administration. For example, it is commonly required that such a material be essentially sterile, e.g., for injectable.

[00221 The term“pharmaceutically acceptable salts” refers to salts that are nontoxic in the amounts and concentrations at which they are administered. The preparation of such salts can facilitate the pharmacological use by altering the physical characteristics of a compound without preventing it from exerting its physiological effect. [0023] The term“pharmaceutically acceptable composition” refers to a

pharmaceutically active compound and one or more pharmaceutically acceptable carriers, excipients, and/or diluents.

[0024] The term“therapeutically effective” or“effective amount” is an amount of a preparation that alone, or together with further doses, and/or in combination with other therapeutic agents produces the desired response. This may involve halting toe progression of the disease or delaying the onset of or preventing the disease or condition from occurring, although it may also imply only slowing of the disease or condition temporarily.

[0025] The term“protein kinase-mediated disease or condition" refers to a disease or condition in which the biological function of a protein kinase affects the development, course, and/or symptoms of the disease or condition.

[0026] The term“mutants" refers to single or multiple amino add changes in a protein as compared to the wild-type protein amino add sequence.

[0027] Throughout the description and claims of this specification, the word "comprise" and variations of the word, such as "comprising” and“comprises,” means“including but not limited to," and is not intended to exclude, for example. other additives, components, integers, or steps. Protein Kinase inhibitors

[0028] The present invention concerns preparation and biological stability of prodrugs of deuteratsd and non-deuterated cyclic chemical compounds and salts thereof active on protein kinases in general, and in particular as inhibitors of protein kinases. These compounds include N-(2-chloro-6~methylphenyt)-2-P- methyl-6-((S)-3-hydroxypyrrolidin-1-yl)pyrimidin-4-yl]amirK)]thiazo|e-5- carboxamide (Compound X), stereoisomers (Compound XI), isomeric mixtures including racemic mixtures (mixture of Compound X and Compound XI), and/or 4-deutero substituted derivatives thereof (Compounds IV and V).

[0029] Compounds IV, V, X, and XI inhibit the activity of one or more protein kinases and are expected to be useful in treating kinase-related diseases or conditions. Compound IV, V, X and XI are disclosed in US 10,479,786 B2, US

10,174,018 B2, US 2019/0071437 A1, and US 2018/0099961 A1.

[0030] Compounds IV, V, X, and Xl provides a method for modulating the activity of a protein kinase selected from the group consisting of ABL, ACK. ARG, BIX, BMX, BRK, B7K, CSK, DDR1, DDR2, EGFR, EPHA1, FGR, FMS, FRK, FYN, HCK, KIT, LCK, LYN, PDGFRa, PDGFRp, SRC, SRM, YES, PIK3CAZP1K3R1 (see US 10,479,786 B2 for more information on the biological properties of these individual protein kinases). Compounds IV, V, X, and XI displayed greater than 50% inhibition at 10 nM of BTK, BMX, ABL, ABL(E255K), SRC, ACK, ARG, BLK,

DDR2, EPHA1, FGR, FMS, FRK, FYNfisoform a), HCK, LCK, LYNa, PDGFRa, PDGFRfr YES, and PIK3CA/PIK3R1. [0031] A further test of biological activity, compounds of the invention were assayed for inhibition of cell growth using diffuse large B-cell lymphoma ceil line SU-DHL-4 and chronic myelogenous leukemia cell line K-562. In this cell-based assay, the IC$o values for Compounds IV, V, X, and XI were all less than 20 nM.

[0032] The protein kinase mediated disease or condition is an autoimmune disease or a cancer. Preferably the autoimmune disease may be at least one of systemic lupus erythematosus (SUE), transplant rejection, multiple sclerosis (MS), systemic sclerosis (SSc), primary Sjogren’s syndrome (pSS), rheumatoid arthritis (RA), and psoriasis. Preferably, the cancer is at least one of Philadelphia chromosome-positive (Ph+) chronic myeloid leukemia (CML), Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL), diffuse large B- ceii lymphoma (DLBCL), chronic lymphocytic leukemia (CLL), follicular lymphoma, marginal zone lymphomas, mantle cell lymphoma (MCL),

Waldenstrom's macroglobu!inemia (WM), T-cell lymphomas, and multiple myeloma.

Administration

[0033] The prodrugs of Compounds IV, V, X, and XI including salts and/or isomeric mixtures thereof, can be formulated as pharmaceutical compositions containing pharmaceutically acceptable excipients this is suitable for oral, transdermai, or parenteral administration. The dosage can be determined by standard procedures taking into account factors such as the released active compound’s IQ»; the biological half-life of tile prodrug and releases active compound; the age, size, and weight of the subject; and the condition associated with the subject. In general, routine experimentation in clinical trials will determine specific ranges far optimal therapeutic effect for each therapeutic agent and each administrative protocol and adminlstiration to specific patients will be adjusted to within effective and safe ranges depending on the patient’s condition and responsiveness to initial administration. However, the ultimate administration protocol will be regulated according to the judgment of the attending clinician considering such factors as age, gender, condition, and size of the patient. Compounds described herein can be administered in single or multiple doses

Preparation of Pnodrugs

[0034] (S)-1 -<6-(5-(2-Chloro-6-methyiphenylcarbamoyl)thiazol-2-ylamino>2- methylpyrimidin-4-yl)pyrrolidin-3-yi acetate (1475-13): An acetate ester prodrug of /V-(2-chloro-6-methylphenyl)-2-[[2-methyl-6-(($)-3-hydroxypyrrolidin-1- yi)pyrimidin-4-yl]amino3thiazole-5-carboxamide (Compound X, 1475-5} was prepared by the synthetic method shown in Scheme 1.

[0035] A similar chemical process can be used to prepare other ester prodrugs of compound X, including the propionate, butanoate, and pentanoate ester prodrugs of compound X. Similarly, compound X can be replaced with its enanttomer /V-(2-chloro-6-methyiphenyi)~2-[[2-methyl-6-((R)-3-hydroxypyrrolidin- 1-yl)pyrimidin-4-y0amino]thiazole-5-carboxamide (compound XI), racemic mixtures (mixtures of Compound X and XI), and/or 4-deutero derivatives

(Compounds IV and V), and racemic mixture of 4-deutero derivatives thereof (mixture of Compounds IV and V).

[0036] (S)-1 -(6-(5-(2-chloro-6-methy}phenylcarbamoyl)thiazol-2-yl)amino-2- methylpyrimidii>4-yl)pynOlidin-3-yi dihydrogen phosphate (1475-16): A

phosphate ester prodrug of yV-(2-chloro-6-methylphenyi)-2-p-methyl-6-((S>3- hydroxypyrrolidin-1 -yl)pyrimidin-4-y(]aminoithiazoie-5-cartroxamide (Compound X) was prepared by the synthetic method shown in Scheme 2.

[0037] A similar chemical process may be used to prepare phosphate prodrugs of compounds IV, V, and XI and racemic mixtures.

[0038] The ester prodrugs of Compounds IV, V, X, and XI will undergo enzymatic and non-enzymatic hydrolysis. Scheme 3 shows the release of Compound X from the phosphate prodrug of Compound X. Similarly, the propionate, butanoate, and pentanoate esters of Compound X can undergo conversion to Compounds X. Compounds IV, V, and XI acetate, propanoate, butanoate, and pentanoate ester prodrugs can also undergo enzymatic and non-enzymatic hydrolysis to liberate Compounds IV, V, and XI.

Scheme 3

[0039] The phosphate prodrugs when administered undergo toe following reaction (Scheme 4) to produce compound X. Similarly, Compounds IV, V, and

XI phosphate prodrugs can also undergo enzymatic and non-enzymatic hydrolysis to liberate Compounds IV, V, and XI.

Scheme4

EXAMPLES

Example 1 :

Preparation of

1-(6-(5-(2-Chloro-6-methylphenylcarbamoyl)thiazol-2- ylamino)-2-methylpyrimidin-4-yl)pyrroiidin-3-yl acetate

[0040] Preparation of (S)-benzyl 3-acetoxypyrrolidine-1 -carboxylate (1475-11): To a stirred mixture of (S)~benzyl 3-hydroxypyrrolidine- 1 -carboxyiate (2 g, 9.04 mmol), pyridine (3 mL, 37,65 mmol, 4 eq), and dichtororhethane (40 mL) was added dropwise at 5°C acetic anhydride (1.7 mL, 17.98 mmol, 2 eq). After completion of the addition, the mixture was stirred at room temperature

overnight TLC analysis (EtOAc-hexanes, 1:4) showed the absence of starting material. The mixture was poured into ice-water and extracted with

dichloromethane (2 x 80 mL). The organic layer was washed with water (60 mL), dried (NaaSO*) and concentrated to give a pale brown liquid (2,7g), which was used in the next step without purification.

[0041] Preparation of (S)-(Pyrrolidin-3-yl) acetate (1475-11-2): A mixture of 1475- 11 (2.7g), methanol (30 mL), and Pd/C (5%) (200mg) was subjected to

hydrogenation using a Ha balloon overnight when TLC showed no starting material. The mixture was then filtered through a layer of ceiite followed by washing with methanol (5 mL). The filtrate and methanol wash were combined and concentrated under reduced pressure to give a pale brown liquid (1.2 g). which was used in next step without purification. [0042] Preparation of (S)-1-(6-(5-(2-Chioro-6-methylphenylcarbamoyl)thiazol-2- ytamino)-2-methylpyrimidin-4-yl)pyrrolidin-3-yl acetate (1475-13): To a mixture of the intermediate A (2-[(6-chloro~2-mefoylpyrimidin"4-yl)amino]~N (2-chloro-6- methylphenyl)thiazole*5-cariX)xamide; 300 mg, 0.76 mmol) and DMSO (10 mL) were added the amine 1475-11-2 ((SHPyrrolidin-S-yl) acetate: 246 mg, ca. 1,9 mmol, 2.5 eq), N ,N-diisopropy!ethy!amine (DIEA) (200 mg, 1.55 mmol, 2 eq), and 4-dimethylaminopyridine (DMAP; 3 mg) at room temperature. The mixture was then stirred at 110°C under nitrogen for 6 h. LC-MS analysis showed the formation of a product peak and disappearance of the peak corresponding to intermediate A. The mixture was cooled to room temperature, followed by addition of water (10 mL) and stirring at room temperature for 15 min. Filtration and washing with water gave foe crude solid product, which was subjected to column purification (EtOAc-hexanes, 1:4 then 1:1) to give 60 mg of 1475-13 9

(acetate ester of compound X) as an off-white solid. LC-MS: 486.90 (M+H).

Example 2

Preparation of (S)-1 -<6-(5-(2-chloro-6-methylphenyicarbamoyi)thia2ml-2-yi)amino-

2-methyl pyrimidin-4-yi)pynOlidin-3-yl dihydrogen phosphate (1475-16)

[0043] To a stirred mixture of the starting material 1475-5 (200 mg, 0.45 mmol) and pyridine (5 mL) at -10°C was added phosphorus oxychloride (0.2 mL, 2.15 mmol, 4.7eq) dropwise. After addition, tire mixture was stirred at 10° to 0°C for 3 h and LC-MS analysis showed the reaction was almost complete. The mixture was cooled to -10°C and water (2 mL) was added dropwise, followed by stirring at 0°C for 30 min. LC-MS analysis confirmed the formation of toe phosphate product The reaction mixture was mixed with 40 mL methylene chloride containing 1 mL diethytamihe, and the resultant mixture was centrifuged at 4000 rpm for 10 min. The resulted pellet from the previous step was re-suspended in 40 mL of distilled water, and the mixture was centrifuged at 4000 rpm for 10 min. The resulted pellet from toe previous step was re-suspended in 40 mL of acetonitrile, and toe mixture was centrifuged at 4000 rpm for 10 min. The resulted petlet was finally re-suspended in 40 mL of methylene chloride, and toe mixture was centrifuged at 4000 rpm for 10 min. The pellet was dried under nitrogen flow to yield approximately 105 mg of the target compound 1475-16 as a solid. LC-MS: 525.2 (M+H); Rt 8.94 min.

Example 3

Biological Stability

[0044] Hydrolysis of 1475-13 (prepared in Example 1) to compound X in human hepatocytes: Cryopreserved pooled human hepatocytes and hepatocyte thawing medium were obtained from Triangle Research Labs (Research Triangle Park, NC, USA). Thawing medium (50 mL) was prewarmed in a 37°C water bath. A vial of hepatocytes was removed from a liquid Na tank and quickly warmed up in a 37°C water bath using slow manual rotation. As soon as toe edge of toe frozen cells were separated from the wall of the vial, toe frozen cells were poured into the prewarmed hepatocyte thawing medium and the remaining cells in foe vial were collected using a pipette. The tube was centrifuged at 100g at 25*C for 5 minutes, foe supernatant was removed, and the cell peilet was resuspended in 8 ml of prewarmed William’s E buffer. The ceil numbers were counted in a hemocytometer. Ceil viability was determined using the trypan blue method. The hepatocyte concentration was adjusted with Wi!fianVs E buffer to 5 million cells/mL.

[0045] To determine the conversion of 1475-13 to compound X, the acetate ester prodrug of compound X, 1475-13 (10 mM), was ihcubated with the 0.5 ml of thawed pooled human hepatocytes at a concentration of 0.5 miilion cells/mL in William's E buffer at 37*C, 5% C0₂ and saturated humidity for 0 (control), 1 and 2 h. At foe end of each incubation time, foe reaction was stopped by the addition of 2x volume of acetonitrile. The mixtures were centrifuged at 4,000 rpm tor 15 min. The resulting supernatant was analyzed using an LC/MSZMS method on an AB Sciex 3000 MS/MS system coupled to a Shimadzu HPLC system. The analytes were separated on a Gemini 5u C18 110A (5 pm, 3.0 mm x 50 mm) reverse phase column from Phenomenex (Torrance, CA) eluted with 2 mM ammonium acetate buffer in water containing 0.1% formic acid and acetonitrile in a step linear gradient.

[0046] FIG 1-3 shows results for incubations for 0 h, 1 h, arid 2 h. After 1 h and 2 h, approximately 50% and 80% of the prodrug 1475-13 was converted to compound X, respectively. In summary, the prodrug 147513 of compound X (N- (2-chloro-6-methylphenyl>2-{[2-methyl-6-(3-hydroxypynOlidin-1-yl)pyrimidin-4- yf]amino]thiazole-5-cartX)xamide) is hydrolyzed by hepatocyte preparations to compound X, a potent inhibitor of protein kinases.

[0047] Conversion of 1475-16 to 14755 in human and mouse hepatocytes;

147516 (10 mM) was incubated with human or mouse primary hepatocytes (0.5 million cells/mL) in William’s E buffer for 4 hours at 37°C under 5% CO2 arid saturated humidity. The reaction was quenched with addition of 3x volume of cold methanol. The mixture was vortexed, and then centrifuged at 4000 rpm for 15 min. The supernatant was subjected for LC-MS/MS analysis, using Sciex API 3000 MS/MS system coupled with Shimadzu 10 series HPLC. The analytes were separated on a Gemini 5u C18 110A (5 pm, 3.0 mm x 50 mm) reverse phase column from Phenomenek (Torrance, CA) eluted with 2 mM ammonium acetate buffer in water containing 0.1% formic add and acetonitrile in a step linear gradient.

[0048] As shown in FIG 4 and 5, 147516 (peak at 59 min) was partially converted to Compound X (peak at 8.4 min) in both human and mouse hepatocytes. In condusion, 147516 as the phosphate prodrug of Compound X can be effectively converted to Compound X in hepatocytes.

Claims

We daim:

1. A compound having the structure

stereoisomers, isomeric mixtures tnduding racemic mixtures, 4-deutero substituted derivatives, and/or a salt thereof; where R is methyl, ethyl, propyl, or butyl.

2. The compound of daim 1, where R is methyl.

3. The compound of daim 1, where R is ethyl.

4. The compound of claim 1 , where R is propyl.

5. The compound of daim 1, where R is butyl.

6. A compound having the structure

stereoisomers, isomeric mixtures including racemic mixtures, 4-deutero substituted derivatives, and/or a salt thereof.

7. A pharmaceutical composition comprising a compound having the following structure

stereoisomers, isomeric mixtures including racemic mixtures, 4-deutero substituted derivatives, and/or a salt thereof, where R is methyl, ethyl, propyl, or butyl.

8. The pharmaceutical composition of claim 7, where R is methyl.

9. The pharmaceutical composition of claim 7, where R is ethyl.

10. The pharmaceutical composition of claim 7, where R is propyl.

11. The pharmaceutical composition of claim 7, where R is butyl.

12. The pharmaceutical composition of claim 7, further containing one or more pharmaceutically acceptable excipients.

13. A pharmaceutical composition comprising a compound having the following structure

14. The pharmaceutical composition of dairri 13, further containing one or more pharmaceutically acceptable excipients.

15. A method of treating a subject suffering from a protein kinase-mediated disease or condition, comprising administering to the subject a therapeutically effective amount of a compound having the following structure

stereoisomers, isomeric mixtures including racemic mixtures, 4-deutero substituted derivatives, and/or a salt thereof, where R is methyl, ethyl, propyl, or butyl, and wherein the protein kinase-mediated disease or condition is at least one of Philadelphia chromosome-positive (Ph+) chronic myeloid leukemia (CML), Philadelphia

chromosome-positive acute lymphoblastic leukemia (Ph+ ALL), diffuse large B-ceil lymphoma (DLBCL), chronic lymphocytic leukemia (CLL), follicular lymphoma, marginal zone lymphomas, mantle cell lymphoma (MCL), Waldenstrom’s macroglobulinemia (WM), T-ce!l lymphomas, multiple myeloma, systemic lupus erythematosus (SLE), transplant rejection, multiple sclerosis (MS), systemic sclerosis (SSc), primary Sjogren's syndrome (pSS), rheumatoid arthritis (RA), and psoriasis.

16. The method of daim 15, where R is methyl.

17. The method of daim 15, where R is ethyl.

18. The method of daim 15, where R is propyl.

19. The method of daim 15, where R is butyl.

20. A method of treating a subject suffering from a protein kinase-mediated disease or condition, comprising administering to the subject a therapeutically effective amount of a compound having the following structure

stereoisomers, isomeric mixtures including racemic mixtures, 4-deutero substituted derivatives, and/or a salt thereof, wherein the protein kinase-mediated disease or condition is at least one of Philadelphia chromosome-positive (Ph+) chronic myeloid leukemia (CML), Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL), diffuse iarge B-cell lymphoma (DLBCL), chronic lymphocytic leukemia (GLL), follicular lymphoma, marginal zone lymphomas, mantle cell lymphoma (MCL),

Waldenstrom’s macroglobulinemia (WM), T-cell lymphomas, multiple myeloma, systemic lupus erythematosus (SLE), transplant rejection, multiple sclerosis (MS), systemic sclerosis (SSc), primary Sjtigren’s syndrome (pSS), rheumatoid arthritis (RA), and psoriasis.