CDK inhibitors and their uses

Abstract

The present disclosure provides compounds and compositions that are CDK inhibitors selective for CDK4 and / or CDK6, and methods of use thereof. [Selection diagram] None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of and priority to U.S. Provisional Patent Application No. 62 / 711,192, filed July 27, 2018, which is incorporated herein by reference in its entirety. [Background technology]

[0002] CDK4 and CDK6 are cyclin-dependent kinases that control the transition between G1 and S phases of the cell cycle. S phase is the period during which cells synthesize new DNA and prepare to divide during the process of mitosis. CDK4 / 6 activity is typically deregulated and overactive in cancer cells. There may be amplification or overexpression of the genes that code for the cyclins or the genes that code for the CDKs themselves. In addition, loss of endogenous inhibitors of CDK4 (also known as INK4 inhibitors) due to gene deletion, mutation, or promoter hypermethylation may also lead to overactivity of CDK4 and CDK6.

[0003] Attempts have been made to prepare compounds that inhibit CDK4 / 6 activity, and some such compounds have been disclosed in the art.However, considering the number of pathological responses mediated by CDK4 / 6, there is still a need for inhibitors of CDK4 / 6 that can be used in the treatment of various conditions, including cancer. Summary of the Invention

[0004] In certain embodiments, a compound of formula (I):

[0005] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, X is independently in each occurrence halo, preferably fluoro; X 1 is O or NRX1 and preferably X 1 is O, R X1 is H or alkyl, preferably lower alkyl; R 1 is alkyl, preferably lower alkyl, and R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, optionally substituted aminoalkyl, or optionally substituted amidoalkyl; R 1 and R 2 together with the carbon atom connecting them together to form an optionally substituted 5- or 6-membered heterocycle (e.g., a pyrrolidine ring, a piperidine ring, a piperazine ring, a pyrrolidine ring, a tetrahydropyran ring, a tetrahydrofuran ring, or a morpholine ring, each of which is optionally substituted). is provided herein.

[0006] In certain embodiments, a compound of formula (Ia):

[0007] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, X is independently in each occurrence halo, preferably fluoro; R 1 is alkyl, preferably lower alkyl, and R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, optionally substituted aminoalkyl, or optionally substituted amidoalkyl; R 1 and R 2together with the carbon atom between which they are linked form an optionally substituted 5- or 6-membered heterocyclic ring. is provided herein.

[0008] In certain embodiments, a compound of formula (Ib):

[0009] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, X is independently in each occurrence halo, preferably fluoro; R X1 is H or alkyl, preferably lower alkyl; R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, optionally substituted aminoalkyl, or optionally substituted amidoalkyl. is provided herein.

[0010] In some embodiments, R 1 is C1-C4-alkyl (e.g., methyl, ethyl).

[0011] In some embodiments, R 2 is optionally substituted C1-C4-alkyl or (CH2) n R 2a [In the formula, R 2ais optionally substituted C1-C4-alkyl, optionally substituted C1-C4-haloalkyl, optionally substituted C2-C4-alkenyl or optionally substituted C1-C4-hydroxyalkyl, optionally substituted C1-C4-alkoxy-C1-C4-alkyl, optionally substituted C1-C4-alkylamino-C1-C4-alkyl or optionally substituted C1-C4-alkylamino-C1-C4-haloalkyl, n is an integer having a value of 1 or 2. It is.

[0012] In certain embodiments, R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl or optionally substituted aminoalkyl.

[0013] For example, in certain embodiments, R 2 is substituted C1-C4-alkyl. In another embodiment, R 2 is methyl, ethyl, propylenyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, (CH2)2OH, -(CH2CH(CH3))OH, (CH2)2O(CH2CH3), -(CH2)2OCH2CH3, -(CH2)2N(H)(CH3), -(CH2)2N(H)(C(CH3)3), -(CH2)2N(H)(C(O)CH3), -(CH2)2N(H)(CH2CH2F), -(CH2)2N(CH3)(CH2CH2F), -(CH2)2N(CH3)2, -(CH2)2N(CH2CH3)2, -(CH2)2N(CH2CH3)2, -(CH2CH(CH3))N(CH3) 2、 -CHC(O)-NHCH 3、 -CH2C(O)-N(CH3)2, -CH2C(O)-N(CH2CH3)2, Or -CH2C(O)-heterocyclyl, such as -CH2C(O)-N-linked heterocyclyl.

[0014] In some embodiments, R 2 (CH2) n C(O)NR 2a R 2b or (CH2) n NR 2a R 2b [In the formula, R 2a and R 2b each independently represents H, alkyl, haloalkyl, alkenyl, (CR c R d ) m OR 2e or -C(O)alkyl; R c , R d and R e are each independently H or alkyl, preferably lower alkyl; n is an integer having a value of 1 or 2; m is an integer having a value from 2 to 5. It is.

[0015] Alternatively, in some embodiments, R 2 (CH2) n C(O)NR 2a R 2b or (CH2) n NR 2a R 2b [In the formula, R 2a and R 2b together with the nitrogen atom which connects them together form an optionally substituted 3- to 6-membered heterocycle; n is an integer having a value of 1 or 2. It is.

[0016] For example, R 2a and R 2b together with the nitrogen atom connecting them,

[0017] [ka] [In the formula, Each R ab is independently halo, hydroxyl, alkyl, haloalkyl, or alkoxy; X 2 O, NR x1 or CR x2 R x3 and R x1 , R x2 and R x3 are each independently H or alkyl, preferably lower alkyl; z is an integer having a value between 0 and 2. may form an optionally substituted heterocycle selected from the following:

[0018] In certain embodiments of the compounds of Formula (I) or Formula (Ia), R 1 and R 2 together with the carbon atoms that connect them together form the structure

[0019] [ka] [In the formula, X 3 is NR Y1a or CR Y1b R Y1c and X 4 is O or CR Y2a R Y2b and R Y1a is H, alkyl, -C(O)R Y1aa or -S(O)alkyl; R Y1aa is alkyl or alkoxy; R Y1b , R Y1c , R Y2a and R Y2b are each independently H or alkyl, preferably lower alkyl. It forms a heterocycle having the following structure:

[0020] In some such embodiments, the heterocycle is

[0021] [ka] is selected from.

[0022] In some embodiments, the compound of formula (I)

[0023] [ka] or a pharma- ceutically acceptable salt thereof.

[0024] In certain embodiments, the present disclosure provides a compound of formula (IVa) or (IVb):

[0025] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, X', independently at each occurrence, is halo, preferably F; R X1' is independently at each occurrence H or lower alkyl; R 1' is C1-C3 alkyl, R 2' is hydroxyalkyl or (CR 2c' 2) n' NR 2a' R 2b' and R 2a' is H, lower alkyl, acyl, or haloalkyl; R 2b' is H, lower alkyl, acyl, or haloalkyl; R 2a' and R 2b' together with the N atom that connects them together form R ab' z' or forming a 4-, 5- or 6-membered heterocyclic ring optionally substituted with R 1' and R 2'together with the C atom(s) which connect them together form a 5- or 6-membered heterocycle which is optionally substituted by acyloxy, R ab' is, when present, independently at each occurrence, halo, hydroxy, lower alkyl, or alkoxy; Each R 2c' is independently H or alkyl, preferably methyl; n' is an integer having a value of 1 or 2; z' is an integer having a value of 0, 1, or 2. and wherein the compound has a CDK4 K i or a pharma- ceutically acceptable salt thereof.

[0026] In some embodiments, the present invention provides a method for the preparation of a compound of formula (V):

[0027] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, R 3a and R 3b together with the nitrogen atom to which they are attached form an optionally substituted [3.3] spiro ring moiety, wherein the optionally substituted [3.3] spiro ring moiety optionally contains at least one additional heteroatom selected from O, S and SO2. With respect to the above, excluding the compound

[0028] [ka] isn't it.

[0029] In some embodiments, the compound has the formula (Va):

[0030] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, R 3a' and R 3b' together with the nitrogen atom to which they are attached

[0031] [ka] forming a structure selected from Each R ab is independently halo, hydroxyl, alkyl, haloalkyl, or alkoxy; z is 0, 1 or 2. has.

[0032] In certain embodiments, R 3a' and R 3b' together with the nitrogen atom to which they are attached,

[0033] [ka] Forming Each R ab are independently haloes, z is 2.

[0034] In some embodiments, R 3a' and R 3b' together with the nitrogen atom to which they are attached,

[0035] [ka] where R ab is fluoro.

[0036] The present disclosure also relates to a composition (e.g., a pharmaceutical composition) comprising a compound of Formula (I), Formula (IVa), Formula (IVb), Formula (V) or Formula (Va) and a carrier (e.g., a pharma- ceutically acceptable carrier, such as a diluent or excipient).

[0037] In certain aspects, the disclosure provides a method of treating a condition or disorder in a subject in need thereof, comprising administering to the subject a compound of Formula (I), Formula (Ia), Formula (Ib), Formula (IVa), Formula (IVb), Formula (V) or Formula (Va) or a pharma- ceutically acceptable salt thereof. Such conditions and disorders include, but are not limited to, cancer, viral infections, inflammatory diseases, cardiovascular diseases, neurodegenerative disorders, glomerulonephritis, myelodysplastic syndromes, myocardial infarction associated with ischemic injury, stroke and reperfusion injury, arrhythmias, atherosclerosis, toxin-induced or alcohol-related liver disease, blood diseases, degenerative diseases of the musculoskeletal system, and eye diseases.

[0038] In certain other aspects, provided herein is a method of sensitizing cancer and / or tumor cells to a chemotherapeutic agent or radiation in a subject in need thereof, comprising administering to the subject an inhibitor of CDK4 and / or CDK6 in an amount sufficient to arrest the cancer and / or tumor cell cycle and thereby sensitize the cancer and / or tumor cells in the mammal to a chemotherapeutic agent or radiation, wherein the inhibitor of CDK4 and / or CDK6 is a compound of Formula (I), Formula (Ia), Formula (Ib), Formula (IVa), Formula (IVb), Formula (V) or Formula (Va), or a pharmaceutically acceptable salt thereof.

[0039] In yet another aspect, provided herein is a method of inhibiting CDK4 and / or CDK6 in a cell, comprising contacting the cell with a compound of Formula (I), Formula (Ia), Formula (Ib), Formula (IVa), Formula (IVb), Formula (V) or Formula (Va) or a pharma- ceutically acceptable salt thereof, such that CDK4 and / or CDK6 enzymes are inhibited in the cell. [Brief description of the drawings]

[0040] [Figure 1] FIG. 1 shows the distribution of abemaciclib IC50 by cancer type. [Diagram 2]FIG. 1 shows the distribution of IC50 of compound A1 (mesylate salt) by cancer type. [Diagram 3] FIG. 1 shows the IC50 distribution of compound A22 (mesylate salt) by cancer type. [Figure 4] FIG. 1 shows the IC50 distribution of compound A23 (mesylate salt) by cancer type. [Diagram 5] FIG. 1 shows the distribution of IC50 of compound A2 (mesylate salt) by cancer type. [Figure 6] Figure 1 shows the relative selectivity of CDK4 / 6 inhibitors palbociclib (200nM), ribociclib (200nM) and abemaciclib (200nM). Compared to palbociclib and ribociclib, abemaciclib hits extra kinases: CDK16, CDK7, DYRK1B, GSK3B, JNK1 / 2 / 3, PIM1, ROCK2, PRKCE, and ribociclib is the cleanest, but hits ULK2. [Figure 7] FIG. 1 shows kinase inhibition screening of certain compounds at 200 nM compared to abemaciclib at 200 nM. [Figure 8] FIG. 1 shows kinase inhibition screening of certain compounds at 2000 nM compared to abemaciclib at 2000 nM. [Figure 9] Figure 1 shows a single dose PK study at 8.3mg / kg investigating PO vs IP and comparative exposure with abemaciclib. This investigative mouse PK study suggested variability in PK for the test compounds. For example, compounds A1 and A2 administered by PO have slow metabolism and longer half-life compared to abemaciclib, and have low Cmax. Compound A49 administered by PO has high Cmax and is rapidly metabolized. Compounds A1 and A2 administered by IP achieved similar bioavailability to abemaciclib. [Figure 10]Figure 1 shows efficacy data from the first ER+ breast cancer cell line xenograft study over 26 days with certain compounds of the present invention. Compound A1 is administered IP at 60mg / kg, suspended for 2 days, and resumed at 20mg / kg QD from day 5 onwards. Compound A22 is administered PO and increased to 120mg / kg from day 12 onwards. Compound A49 is administered PO and administration is stopped on day 19. ZR751 cell line is used in this study. [Figure 11] Figure 1 shows efficacy data from the first ER+ breast cancer cell line xenograft study over 26 days with certain compounds of the present invention. Compound A1 is administered IP at 60mg / kg, suspended for 2 days, and resumed at 20mg / kg QD from day 5 onwards. Compound A22 is administered PO and increased to 120mg / kg from day 12 onwards. Compound A49 is administered PO and administration is stopped on day 19. ZR751 cell line is used in this study. [Figure 12A] FIG. 1 shows results from the first ER+ breast cancer cell line xenograft study. [Figure 12B] FIG. 1 shows results from the first ER+ breast cancer cell line xenograft study. [Figure 13] FIG. 1 shows the results of a follow-up PK study for certain compounds of the invention, each dosed at 100 mg / kg by PO. [Figure 14] FIG. 1 shows an exemplary treatment regimen for a second ER+ breast cancer cell line xenograft study. The ZR751 cell line was used in this study. [Figure 15] FIG. 13 shows results of a second xenograft efficacy study at day 22. [Figure 16] FIG. 13 shows results of a second xenograft efficacy study at day 22. [Figure 17] FIG. 1 depicts PK results from a second xenograft efficacy study determined from plasma collected at 240 and 1,440 minutes post-dose on study day 10. [Figure 18A] FIG. 1 shows comparative AUC measurements for compounds A2, A1, A23 and abemaciclib at doses of 100-1000 mg / kg. [Figure 18B] FIG. 1 shows comparative PK curves for compounds A2, A1, A23 and abemaciclib at doses of 100-1000 mg / kg. [Figure 19] FIG. 1 shows an exemplary design for a 14-day dose tapering study. [Figure 20] 1 shows the results of a 14-day dose tapering study of Compound A2. The maximum tolerated dose of Compound A2 was between about 278 and about 399 mg / kg QD. [Figure 21] 1 shows the results of a 14-day dose tapering study of Compound A2. The maximum tolerated dose of Compound A2 was between about 278 and about 399 mg / kg QD. [Figure 22] 1 shows the results of a 14-day dose tapering study of Compound A1. The maximum tolerated dose of Compound A1 was less than about 400 mg / kg QD. [Diagram 23] 1 shows the results of a 14-day dose tapering study of Compound A1. The maximum tolerated dose of Compound A1 was less than about 400 mg / kg QD. [Figure 24]

[0023] Figure 1 shows the results of a 14-day dose tapering study on Compound A23. The maximum tolerated dose of Compound A23 was approximately 200 mg / kg QD. [Diagram 25]

[0023] Figure 1 shows the results of a 14-day dose tapering study of Compound A23. The maximum tolerated dose of Compound A23 was approximately 200 mg / kg QD. [Figure 26] 1 shows the results of a 14-day dose tapering study of abemaciclib. The maximum tolerated dose of abemaciclib was between about 100 and about 150 mg / kg QD. [Figure 27] 1 shows the results of a 14-day dose tapering study of abemaciclib. The maximum tolerated dose of abemaciclib was between about 100 and about 150 mg / kg QD. [Figure 28] FIG. 1 shows an exemplary treatment regimen for a xenograft study involving 8 mice per experimental group. Blood was collected on the first and last treatment days. [Figure 29]FIG. 1 shows xenograft efficacy data over the 26 day study and on the final day (day 26), respectively. [Diagram 30] FIG. 1 shows xenograft efficacy data over the 26 day study and on the final day (day 26), respectively. [Diagram 31] 1 is a graphical representation of animal weight data over the 26 day period of the xenograft study. [Diagram 32] FIG. 1 shows PK data for compounds A1, A2, A23, and abemaciclib at the day 25 blood draw of the xenograft study. [Diagram 33] FIG. 1 shows accumulation of compounds of the invention versus abemaciclib over a 26 day xenograft study. [Diagram 34] FIG. 13 tabulates body weight data over 25 days of treatment in the xenograft study. [Figure 35A] FIG. 13 tabulates animal weight data on day 26 (final day) of the xenograft study. [Figure 35B] FIG. 13 tabulates animal weight data on day 26 (final day) of the xenograft study. [Diagram 36] FIG. 1 shows a summary of results from a xenograft efficacy study. [Figure 37] FIG. 1 shows a summary of the log difference in mean IC50 of certain compounds between resistant and sensitive cell lines. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0041] Abemaciclib (brand name Verzenio(TM)):

[0042] [ka] Abemaciclib inhibits the enzymes cyclin-dependent kinase 4 (CDK4) and cyclin-dependent kinase 6 (CDK6). These enzymes are involved in phosphorylating and activating retinoblastoma proteins that play a role in cell cycle progression from G1 (first gap) to S (synthesis) phase. Blocking this pathway prevents cells from progressing to S phase, thereby inducing apoptosis (cell death). The compounds disclosed herein share certain structural features of abemaciclib and may also act as selective CDK inhibitors for CDK4 and / or CDK6.

[0043] definition Unless otherwise specified herein, scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. In general, the nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics, and protein and nucleic acid chemistry described herein are those well known and commonly used in the art.

[0044] The methods and techniques of the present disclosure, unless otherwise indicated, are generally performed according to conventional methods well known in the art and as described in the various general and more specific references cited and discussed throughout this specification. See, for example, "Principles of Neural Science", McGraw-Hill Medical, New York, NY (2000); Motulsky, "Intuitive Biostatistics", Oxford University Press, Inc. (1995); Lodish et al., "Molecular Cell Biology, 4th ed.", WH Freeman & Co., New York (2000); Griffiths et al., "Introduction to Genetic Analysis, 7th ed.", WH Freeman & Co., NY (1999); and Gilbert et al., "Developmental Biology, 6th ed.", Sinauer Associates, Inc., Sunderland, MA (2000).

[0045] Chemical terms used herein, unless otherwise defined herein, are used according to conventional usage in the art as exemplified by "The McGraw-Hill Dictionary of Chemical Terms", Parker S., Ed., McGraw-Hill, San Francisco, CA (1985).

[0046] All of these and any other publications, patents and published patent applications referenced herein are specifically incorporated herein by reference. In case of conflict, the present specification, including its specific definitions, will control.

[0047] "Patient," "subject," or "individual" are used interchangeably and refer to either a human or non-human animal. These terms include mammals, such as humans, primates, livestock animals (including cows, pigs, etc.), pet animals (e.g., dogs, cats, etc.), and rodents (e.g., mice and rats).

[0048] "Treating" a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. As used herein, and as is well understood in the art, "treatment" is an approach to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results may include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, whether detectable or undetectable, reduction in the extent of disease, stabilization of the disease state (i.e., not worsening), prevention of disease spread, delay or slowing of disease progression, improvement or palliation of the disease state, and remission (whether partial or total). "Treatment" may also mean prolonging survival compared to expected survival if not treated.

[0049] The term "preventing" is art-recognized and well understood in the art when used in connection with a condition, e.g., local recurrence (e.g., pain), a disease, e.g., cancer, a complex syndrome, e.g., heart failure, or any other medical condition, and includes administration of a composition that reduces the frequency or delays the onset of symptoms of a medical condition in a subject compared to subjects not receiving the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment compared to an untreated control population by a statistically and / or clinically significant amount, and / or delaying the appearance of detectable cancerous growths in a treated population relative to an untreated control population.

[0050] "Administering" or "administration" of a substance, compound or agent to a subject can be performed using one of a variety of methods known to those skilled in the art. For example, the compound or agent can be administered intravenously, intraarterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (e.g., by absorption through the skin's channels). The compound or agent can also be suitably introduced by rechargeable or biodegradable polymeric or other devices, such as patches and pumps, or formulations that provide sustained, slow or controlled release of the compound or agent. Administration can also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0051] Suitable methods for administering a substance, compound or agent to a subject will also depend, for example, on the age and / or physical condition of the subject, and the chemical and biological properties of the compound or agent (e.g., solubility, digestibility, bioavailability, stability and toxicity). In some embodiments, the compound or agent is orally administered to the subject, for example, by ingestion. In some embodiments, the orally administered compound or agent is in a sustained or timed release formulation or is administered using such a sustained or timed release device.

[0052] As used herein, the phrase "co-administration" refers to any form of administration of two or more different therapeutic agents, such that a second agent is administered while a previously administered therapeutic agent is still effective in the body (e.g., two agents are effective in a patient at the same time, which may include a synergistic effect of the two agents). For example, different therapeutic compounds can be administered either simultaneously or sequentially, either in the same formulation or in separate formulations. Thus, an individual undergoing such treatment can benefit from the combined effect of the different therapeutic agents.

[0053] The term "acyl" is art-recognized and refers to a group represented by the general formula: hydrocarbylC(O)-, preferably alkylC(O)-.

[0054] The term "acylamino" is art recognized and refers to an amino group substituted with an acyl group and can be represented, for example, by the formula: hydrocarbylC(O)NH-.

[0055] The term "acyloxy" is art-recognized and refers to a group represented by the general formula: hydrocarbylC(O)O-, preferably alkylC(O)O-.

[0056] The term "alkoxy" refers to an alkyl group, preferably a lower alkyl group, having an oxygen attached to an alkyl group, preferably a lower alkyl group. Representative alkoxy groups include methoxy, trifluoromethoxy, ethoxy, propoxy, tert-butoxy, and the like.

[0057] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group and can be represented by the general formula: alkyl-O-alkyl.

[0058] The term "alkenyl" as used herein refers to an aliphatic group containing at least one double bond, and is intended to include both "unsubstituted alkenyl" and "substituted alkenyl", the latter of which refers to an alkenyl moiety having a substituent replacing a hydrogen on one or more carbons of the alkenyl group. Such substituents may occur on one or more carbons that are or are not included in one or more double bonds. Furthermore, such substituents include all of those contemplated for alkyl groups, as described below, except where stability prohibits their use. For example, substitution of alkenyl groups with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0059] An "alkyl" group or "alkane" is a straight or branched chain non-aromatic hydrocarbon that is fully saturated. Typically, a straight or branched chain alkyl group has from 1 to about 20 carbon atoms, preferably from 1 to about 10 carbon atoms, unless otherwise specified. Examples of straight and branched chain alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl and octyl. C1-C6 straight or branched chain alkyl groups are also referred to as "lower alkyl" groups.

[0060] Furthermore, the term "alkyl" (or "lower alkyl"), as used throughout the specification, examples, and claims, is intended to include both "unsubstituted alkyl" and "substituted alkyl," the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents, unless otherwise specified, include, for example, halogen (e.g., fluoro), hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. In preferred embodiments, the substituents on the substituted alkyl are selected from C1-C6 alkyl, C3-C6 cycloalkyl, halogen, carbonyl, cyano, or hydroxyl. In more preferred embodiments, the substituents on the substituted alkyl are selected from fluoro, carbonyl, cyano, or hydroxyl. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, where appropriate. For example, substituents on the substituted alkyl include substituted and unsubstituted amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl, and sulfonates), and silyl groups, as well as ethers, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, and the like. Exemplary substituted alkyls are described below. Cycloalkyls can be further substituted with alkyls, alkenyls, alkoxy, alkylthio, aminoalkyls, carbonyl-substituted alkyls, -CF3, -CN, and the like.

[0061] The term “C x ~C y" when used in conjunction with a chemical moiety, such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups that contain x to y carbons in the chain. For example, the term "C x ~C y "Alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group, including straight chain alkyl and branched chain alkyl groups, containing x to y carbons in the chain, including haloalkyl groups. Preferred haloalkyl groups include trifluoromethyl, difluoromethyl, 2,2,2-trifluoroethyl, and pentafluoroethyl. CO alkyl refers to a hydrogen when the group is in a terminal position, and a bond when the group is internal. The term "C2-C y Alkenyl" and "C2-C y "Alkynyl" refers to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double or triple bond respectively.

[0062] The term "alkylamino," as used herein, refers to an amino group substituted with at least one alkyl group.

[0063] The term "alkylthio," as used herein, refers to a thiol group substituted with an alkyl group and can be represented by the general formula: alkylS-.

[0064] The term "alkynyl" as used herein refers to an aliphatic group containing at least one triple bond, and is intended to include both "unsubstituted alkynyl" and "substituted alkynyl", the latter of which refers to an alkynyl moiety having a substituent replacing a hydrogen on one or more carbons of the alkynyl group. Such a substituent may be present on one or more carbons that are or are not included in one or more triple bonds. Furthermore, such substituents include all of those contemplated for alkyl groups, as described above, except where stability prohibits their use. For example, substitution of alkynyl groups with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

[0065] The term "amide," as used herein, refers to the group:

[0066] [ka] [In the formula, each R A independently represent hydrogen or a hydrocarbyl group, or two R A together with the N atom to which they are attached complete a heterocycle having 4-8 atoms in the ring structure. Refers to...

[0067] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines, and their salts, e.g.,

[0068] [ka] [In the formula, each R A independently represent hydrogen or a hydrocarbyl group, or two R A together with the N atom to which they are attached complete a heterocycle having 4-8 atoms in the ring structure. It refers to a portion that can be represented by:

[0069] The term "aminoalkyl," as used herein, refers to an alkyl group substituted with an amino group.

[0070] The term "aryl" as used herein includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. Preferably, the ring is 6 or 10-membered, more preferably 6-membered. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is aromatic, and the other cyclic rings can be, for example, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

[0071] A "cycloalkyl" group is a fully saturated cyclic hydrocarbon. "Cycloalkyl" includes monocyclic and bicyclic rings. Typically, a monocyclic cycloalkyl group has from 3 to about 10 carbon atoms, more typically from 3 to 8 carbon atoms, unless otherwise specified. The second ring of a bicyclic cycloalkyl can be selected from saturated, unsaturated, and aromatic rings. Cycloalkyl includes bicyclic molecules in which one, two, or three or more atoms are shared between the two rings (e.g., fused bicyclic compounds, bridged bicyclic compounds, and spirocyclic compounds).

[0072] The term "fused bicyclic compound" refers to a bicyclic molecule in which two rings share two adjacent atoms. In other words, the rings share one covalent bond, i.e., the so-called bridgehead atoms are directly bonded (e.g., α-thujene and decalin). For example, in a fused cycloalkyl, each of the rings shares two adjacent atoms with the other ring, and the second ring of the fused bicyclic cycloalkyl can be selected from saturated, unsaturated, and aromatic rings. A "cycloalkenyl" group is a cyclic hydrocarbon containing one or more double bonds.

[0073] The term "bridged bicyclic compound" refers to a bicyclic molecule in which two rings share three or more atoms and a bridge containing at least one atom separates the two bridgehead atoms. For example, norbornane, also known as bicyclo[2.2.1]heptane, can be thought of as a pair of cyclopentane rings each sharing three of their five carbon atoms.

[0074] The term "spirocyclic compound" refers to a bicyclic molecule in which the two rings have only one single atom in common, the spiroatom.

[0075] The terms "halo" and "halogen" as used herein mean halogens and include chloro, fluoro, bromo and iodo.

[0076] The term "heteroalkyl," as used herein, refers to a saturated or unsaturated chain of carbon atoms and at least one heteroatom, where no two heteroatoms are adjacent.

[0077] The terms "heteroaryl" and "hetaryl" include substituted or unsubstituted aromatic monocyclic structures, preferably 5-7 membered rings, more preferably 5-6 membered rings, which ring structures contain at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heteroaryl" and "hetaryl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is heteroaromatic, and the other cyclic rings can be, for example, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.

[0078] The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen and sulfur.

[0079] The terms "heterocyclyl", "heterocycle" and "heterocyclic" refer to substituted or unsubstituted non-aromatic ring structures, preferably 3-10 membered rings, more preferably 3-7 membered rings, which ring structures contain at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1 or 2 heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is heterocyclic, for example, the other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and / or heterocyclyl. Heterocyclyl groups include, for example, pyrrolidine, piperidine, piperazine, pyrrolidine, tetrahydropyran, tetrahydrofuran, morpholine, lactones, lactams, and the like.

[0080] The term "hydroxyalkyl," as used herein, refers to an alkyl group substituted with a hydroxy group.

[0081] The term "lower" when used in conjunction with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, is meant to include groups in which there are 10 or fewer non-hydrogen atoms in the substituent, preferably 6 or fewer. "Lower alkyl" refers, for example, to alkyl groups containing 10 or fewer, preferably 6 or fewer carbon atoms. In certain embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, respectively, whether they appear alone or in combination with other substituents, for example, in the recitation of hydroxyalkyl and aralkyl (where, for example, atoms in aryl groups are not counted when counting carbon atoms in an alkyl substituent).

[0082] The term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbons of the backbone. It will be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is in accordance with the permissible valences of the substituted atom and substituent, as well as that the substitution results in a stable compound that does not spontaneously undergo transformation, for example, by rearrangement, cyclization, elimination, and the like. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. The substituents may include any of the substituents described herein, such as halogen, hydroxyl, carbonyl (e.g., carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (e.g., thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. In preferred embodiments, the substituents on the substituted alkyl are selected from C1-C6 alkyl, C3-C6 cycloalkyl, halogen, carbonyl, cyano, or hydroxyl. In more preferred embodiments, the substituents on the substituted alkyl are selected from fluoro, carbonyl, cyano, or hydroxyl. It will be appreciated by those of skill in the art that, where appropriate, the substituents may themselves be substituted. Unless specifically stated as "unsubstituted," references herein to chemical moieties are understood to include substituted variants. For example, reference to an "aryl" group or moiety implicitly includes both substituted and unsubstituted variants.

[0083] "Protecting group" refers to a group of atoms that, when attached to a reactive functional group in a molecule, masks, reduces or prevents the reactivity of the functional group. Typically, a protecting group can be selectively removed as desired during the synthetic process. Examples of protecting groups can be found in Greene and Wuts, Protective Groups in Organic Chemistry, 3 rd Ed., 1999, John Wiley & Sons, NY, and Harrison et al., Compendium of Synthetic Organic Methods, Vols. 1-8, 1971-1996, John Wiley & Sons, NY. Representative nitrogen protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilyl-ethanesulfonyl ("TES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl ("FMOC"), nitro-veratryloxycarbonyl ("NVOC"), and the like. Representative hydroxyl protecting groups include, but are not limited to, those in which the hydroxyl group is acylated (esterified) or alkylated, such as benzyl and trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers (e.g., TMS or TIPS groups), glycol ethers, such as ethylene glycol and propylene glycol derivatives, and allyl ethers.

[0084] The term "modulate" as used herein includes inhibiting or suppressing a function or activity (eg, cell proliferation), as well as enhancing a function or activity.

[0085] The phrase "pharmacologically acceptable" is art-recognized. In certain embodiments, this term includes compositions, excipients, adjuvants, polymers, and other materials and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, and are commensurate with a reasonable benefit / risk ratio.

[0086] "Pharmaceutically acceptable salt" or "salt" is used herein to refer to acid addition salts or base addition salts that are suitable or compatible with the treatment of patients.

[0087] The term "pharmaceutically acceptable acid addition salts" as used herein means any non-toxic organic or inorganic salt of any base compound disclosed herein. Exemplary inorganic acids that form suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Exemplary organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid, and salicylic acid, as well as sulfonic acids such as p-toluenesulfonic acid and methanesulfonic acid. Either mono- or di-acid salts can be formed, and such salts can exist in either hydrated, solvated, or substantially anhydrous form. In general, acid addition salts of the compounds disclosed herein are more soluble in water and various hydrophilic organic solvents, and generally exhibit higher melting points compared to their free base forms. The selection of an appropriate salt will be known to one of skill in the art. Other pharmaceutically unacceptable salts, such as oxalates, may be used, for example, in the isolation of compounds of the invention for laboratory use or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0088] The term "pharmaceutically acceptable base addition salt" as used herein means any non-toxic organic or inorganic base addition salt of any acid compound of the present invention, or any of its intermediates. Exemplary inorganic bases that form suitable salts include lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, or barium hydroxide. Exemplary organic bases that form suitable salts include aliphatic, alicyclic, or aromatic organic amines, such as methylamine, trimethylamine, and picoline, or ammonia. The selection of appropriate salts will be known to those skilled in the art.

[0089] Many of the compounds useful in the methods and compositions of the present disclosure have at least one stereocenter in their structure. This stereocenter can be in the R or S configuration, and the R and S designations are used according to the rules set forth in Pure Appl. Chem. (1976), 45, 11-30. The present disclosure contemplates all stereoisomeric forms of the compounds, salts, prodrugs, or mixtures thereof, including enantiomeric and diastereoisomeric forms (including all possible mixtures of stereoisomers). See, for example, WO 01 / 062726.

[0090] Furthermore, certain compounds containing alkenyl groups can exist as Z (zusammen) or E (entgegen) isomers, and in each case the present disclosure includes both mixtures and the separate individual isomers.

[0091] Some of the compounds may also exist in tautomeric forms. Such forms, although not explicitly shown in the formulae depicted herein, are intended to be included within the scope of the present disclosure.

[0092] "Prodrug" or "Pharmaceutically acceptable prodrug" refers to a compound that is metabolized, e.g., hydrolyzed or oxidized, in the host after administration to form a compound of the present disclosure (e.g., a compound of the present invention). Typical examples of prodrugs include compounds that have a biologically labile or cleavable (protecting) group on a functional moiety of an active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated, acylated, deacylated, phosphorylated, or dephosphorylated to generate an active compound. Examples of prodrugs that use esters or phosphoramidates as biologically labile or cleavable (protecting) groups are disclosed in U.S. Patent Nos. 6,875,751, 7,585,851, and 7,964,580, the disclosures of which are incorporated herein by reference. The prodrugs of the present disclosure are metabolized to generate a compound of the present invention or a pharmaceutically acceptable salt thereof. The present disclosure includes within its scope prodrugs of the compounds described herein. Conventional procedures for the selection and preparation of suitable prodrugs are described, for example, in “Design of Prodrugs” Ed. H. Bundgaard, Elsevier, 1985.

[0093] The phrase "pharmaceutically acceptable carrier" as used herein means a pharma- ceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, useful in formulating a drug for pharmaceutical or therapeutic use.

[0094] As used herein, "inhibitors of CDK4 / 6" or "CDK4 / 6 inhibitor therapy" refers to compounds or compositions that inhibit the activity of CDK4 / 6, e.g., to phosphorylate serine or threonine residues on proteins, or that inhibit the interaction of CDK4 / 6 with other proteins that may be in a signaling pathway.

[0095] As used herein, "sensitive to a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor" or "CDK4 / 6i-sensitive cancer" refers to a cell or cancer that has reduced proliferation in the presence of a CDK4 / 6 inhibitor compared to the absence of such an inhibitor. Sensitivity may refer to the cytotoxic or cytostatic effect of a CDK4 / 6 inhibitor on the cell. It is contemplated that a sensitive cell line may have a 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25-fold or more change in proliferation rate in the presence of a CDK4 / 6 inhibitor. Sensitivity may also be measured by changes in the genomic sequence or copy number of a gene, an increase or decrease in specific protein expression or mRNA expression, or other measurements disclosed herein that are indicative of sensitivity.

[0096] The term "response to CDK4 and / or CDK6 inhibitors" relates to any response of a hyperproliferative disorder (e.g., cancer) to an agent that inhibits CDK4 or CDK6, preferably to the change in tumor mass and / or volume after the initiation of chemotherapy. Hyperproliferative disorder response may be evaluated, for example, for efficacy or in a neoadjuvant or adjuvant setting, where the size of the tumor after systemic intervention can be measured by CT, PET, mammogram, ultrasound or palpation and compared to the initial size and dimensions. Response may also be evaluated by caliper measurement or pathological examination of the tumor after biopsy or surgical resection. Response may be recorded in a quantitative manner, such as the percentage change in tumor volume, or in a qualitative manner, such as "pathological complete response" (pCR), "clinical complete response" (cCR), "clinical partial response" (cPR), "clinical stable disease" (cSD), "clinical progressive disease" (cPD) or other qualitative criteria. The assessment of hyperproliferative disorder response may be performed early, for example, hours, days, weeks, or preferably months, after the initiation of neoadjuvant or adjuvant therapy. A typical end point for response assessment is the end of neoadjuvant chemotherapy or the surgical removal of residual tumor cells and / or tumor bed. This is typically 3 months after the initiation of neoadjuvant therapy. In some embodiments, the clinical efficacy of the therapeutic treatment described herein can be determined by measuring the clinical benefit rate (CBR). The clinical benefit rate is measured by determining the percentage of patients in complete remission (CR), the number of patients in partial remission (PR), and the number of patients with stable disease (SD) at least 6 months after the end of treatment. An abbreviation of this formula is CBR=CR+PR+SD over 6 months. In some embodiments, the CBR for a particular cancer treatment regimen is at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or more.Further criteria for assessing response to cancer therapy relate to "survival", including all of the following: survival to death, also known as overall survival (wherein the death may be cause-unrelated or tumor-related); "recurrence-free survival" (wherein the term recurrence includes both local and distant recurrence); metastasis-free survival; disease-free survival (wherein the term disease includes cancer and its associated diseases). The length of survival can be calculated by reference to a defined starting point (e.g., time of diagnosis or start of treatment) and an end point (e.g., death, recurrence or metastasis). In addition, criteria for efficacy of treatment can be expanded to include response to chemotherapy, probability of survival, probability of metastasis within a given period, and probability of tumor recurrence. For example, to determine an appropriate threshold, a particular cancer treatment regimen can be administered to a population of subjects, and the results can be correlated with biomarker measurements determined before administering any cancer therapy. The outcome measurement can be a pathological response to therapy given in a neoadjuvant setting. Alternatively, outcome measures, such as overall survival and disease-free survival, can be monitored for a period of time for subjects after cancer treatment for which biomarker measurements are known. In certain embodiments, the dose administered is a standard dose known in the art for cancer therapeutics. The period for which a subject is monitored can vary. For example, a subject can be monitored for at least 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 45, 50, 55, or 60 months.

[0097] The term "survival" includes all of the following: survival until death, also known as overall survival (wherein the death may be cause-unrelated or tumor-related); "recurrence-free survival" (wherein the term recurrence includes both local and distant recurrence); metastasis-free survival; disease-free survival (wherein the term disease includes cancer and its associated diseases). The length of survival can be calculated by reference to a defined starting point (e.g., time of diagnosis or treatment initiation) and an end point (e.g., death, recurrence, or metastasis). In addition, the criteria for efficacy of treatment can be expanded to include response to chemotherapy, probability of survival, probability of metastasis within a given period, and probability of tumor recurrence.

[0098] As used herein, "resistant to a cyclin-dependent kinase 4 / 6 (CDK4 / 6) inhibitor" or "CDK4 / 6i-resistant cancer" refers to a cell or cancer that has normal (or baseline) growth in the presence of a CDK4 / 6 inhibitor, substantially similar to that in the absence of such inhibitor. Resistance can be measured by the relative maintenance of cell proliferation rate in the presence of a CDK4 / 6 inhibitor, or by changes in the genomic sequence or copy number of a gene, an increase or decrease in specific protein expression or mRNA expression, or other measurements disclosed herein that are indicative of resistance.

[0099] The term "sensitize" refers to altering cancer or tumor cells in a manner that allows for more effective treatment of the associated cancer with cancer therapy (e.g., anti-immune checkpoint therapy, chemotherapy and / or radiation therapy). In some embodiments, normal cells are not affected by immune checkpoint therapy to the extent that they are unduly damaged. Increased susceptibility or decreased susceptibility to a therapeutic treatment is measured according to methods known in the art for the particular treatment and described herein below, including, but not limited to, cell proliferation assays (Tanigawa N, Kern DH, Kikasa Y, Morton DL, Cancer Res 1982; 42: 2159-2164), and cell death assays (Weisenthal LM, Shoemaker RH, Marsden JA, Dill PL, Baker JA, Moran EM, Cancer Res 1984; 94: 161-173; Weisenthal LM, Lippman ME, Cancer Treat Rep 1985; 69: 615-632; Weisenthal LM, In: Kaspers GJL, Pieters R, Twentyman PR, Weisenthal LM, Veerman AJP, eds. Drug Resistance in Leukemia and Lymphoma. Langhorne, PA: Harwood J. Med. 1999; 10: 1111-1123 ... Academic Publishers, 1993: 415-432; Weisenthal LM, Contrib Gynecol Obstet 1994; 19: 82-90). Sensitivity or resistance can also be measured in animals by measuring the reduction in tumor size over a period of time, e.g., 6 months in humans and 4-6 weeks in mice.A composition or method sensitizes a response to a therapeutic treatment when the increase in treatment sensitivity or the reduction in resistance is 25% or more, e.g., 30%, 40%, 50%, 60%, 70%, 80% or more, to 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 15-fold, 20-fold or more, compared to the treatment sensitivity or resistance in the absence of such composition or method. Determining sensitivity or resistance to a therapeutic treatment is routine in the art and within the skill of the artisan. It should be understood that any method described herein for enhancing the effectiveness of cancer treatment is equally applicable to the method of sensitizing hyperproliferative or other cancerous cells (e.g., resistant cells) to cancer treatment.

[0100] Compounds of the Invention The present disclosure provides a compound of formula (I):

[0101] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, X 1 is O or NR X1 and R X1 is H or alkyl, preferably lower alkyl; R 1 is alkyl, preferably lower alkyl, and R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, optionally substituted aminoalkyl, or optionally substituted amidoalkyl; R 1 and R 2 together with the carbon atom between which they are linked form an optionally substituted 5- or 6-membered heterocyclic ring. to provide.

[0102] In certain embodiments, R 2is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, or optionally substituted aminoalkyl. In certain embodiments, the formula (Ia):

[0103] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, X is independently in each occurrence halo, preferably fluoro; R 1 is alkyl, preferably lower alkyl, and R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, optionally substituted aminoalkyl, or optionally substituted amidoalkyl; R 1 and R 2 together with the carbon atom between which they are linked form an optionally substituted 5- or 6-membered heterocyclic ring. is provided herein.

[0104] In certain embodiments, R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl or optionally substituted aminoalkyl.

[0105] In certain embodiments, a compound of formula (Ib):

[0106] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, X is independently in each occurrence halo, preferably fluoro; R X1 is H or alkyl, preferably lower alkyl; R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, optionally substituted aminoalkyl, or optionally substituted amidoalkyl. is provided herein.

[0107] In certain embodiments, R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl or optionally substituted aminoalkyl.

[0108] In some embodiments, the compound of formula (I) has the formula (II):

[0109] [ka] or a pharma- ceutically acceptable salt thereof.

[0110] In some such embodiments, the compound of formula (II) has the formula (IIa):

[0111] [ka] Alternatively, the compound of formula (II) has the structure of formula (IIb):

[0112] [ka] or a pharma- ceutically acceptable salt thereof.

[0113] In some embodiments, the compound of formula (I) has formula (III):

[0114] [ka] or a pharma- ceutically acceptable salt thereof.

[0115] In some such embodiments, the compound of Formula (III) has the formula (IIIa):

[0116] [ka] Alternatively, the compound of formula (III) has the structure of formula (IIIb):

[0117] [ka] or a pharma- ceutically acceptable salt thereof.

[0118] In certain embodiments of compounds of Formula (I), (Ia), (Ib), (III), (IIIa) and (IIIb), R X1 is H or methyl. In a preferred embodiment, R X1 is H.

[0119] In some embodiments of the disclosed compounds, R 1 is C1-C4-alkyl. In some such embodiments, R 1 is methyl or ethyl. In certain embodiments, R 1 is methyl.

[0120] In some embodiments of the disclosed compounds, R 2 is optionally substituted C1-C4-alkyl or (CH2) n R 2a where R 2ais optionally substituted C1-C4-alkyl, optionally substituted C1-C4-haloalkyl, optionally substituted C2-C4-alkenyl, or optionally substituted C1-C4-hydroxyalkyl, optionally substituted C1-C4-alkoxy-C1-C4-alkyl, optionally substituted C1-C4-alkylamino-C1-C4-alkyl, or optionally substituted C1-C4-alkylamino-C1-C4-haloalkyl, and n is an integer having a value of 1 or 2. In some such embodiments, R 2 is substituted C1-C4-alkyl. In another embodiment, R 2 are methyl, ethyl, propylenyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, (CH2)2OH, -(CH2CH(CH3))OH, (CH2)2O(CH2CH3), -(CH2)2OCH2CH3, -(CH2)2N(H)(CH3), -(CH2)2N(H)(C(CH3)3), -(CH2)2N(H)(C(O)CH3), -(CH2)2N(H)(CH2CH2F), -(CH2)2N(CH3)(CH2CH2F), -(CH2)2N(CH3)2, -(CH2)2N(CH2CH3)2, -(CH2)2N(CH2CH3)2, -(CH2CH(CH3))N(CH3)2, -CH2C(O)-NHCH 3、 -CH2C(O)-N(CH3)2, -CH2C(O)-N(CH2CH3)2, or -CHC(O)-heterocyclyl, e.g., -CHC(O)-N-linked heterocyclyl. 2is methyl, ethyl, propylenyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, (CH2)2OH, -(CH2CH(CH3))OH, (CH2)2O(CH2CH3), -(CH2)2OCH2CH3, -(CH2)2N(H)(CH3), -(CH2)2N(H)(C(CH3)3), -(CH2)2N(H)(C(O)CH3), -(CH2)2N(H)(CH2CH2F), -(CH2)2N(CH3)(CH2CH2F), -(CH2)2N(CH3)2, -(CH2)2N(CH2CH3)2, -(CH2)2N(CH2CH3)2, or -(CH2CH(CH3))N(CH3)2 It is.

[0121] In other embodiments of the disclosed compounds, R 2 (CH2) n C(O)NR 2a R 2b where R 2a and R 2b each independently represents H, alkyl, haloalkyl, alkenyl, (CR c R d ) m OR e or -C(O)alkyl, R c , R d and R e are each independently H or alkyl, preferably lower alkyl; n is an integer having a value of 1 or 2; and m is an integer having a value of 2-5.

[0122] In other embodiments of the disclosed compounds, R 2 (CH2) n NR 2a R 2b where R 2a and R 2b each independently represents H, alkyl, haloalkyl, alkenyl, (CR c R d ) m OR e or -C(O)alkyl, R c , R d and R eare each independently H or alkyl, preferably lower alkyl; n is an integer having a value of 1 or 2; and m is an integer having a value of 2-5.

[0123] In still other embodiments of the disclosed compounds, R 2 (CH2) n C(O)NR 2a R 2b where R 2a and R 2b together with the nitrogen atom which connects them together to form an optionally substituted 3- to 6-membered heterocycle, and n is an integer having a value of 1 or 2. In some such embodiments, R 2a and R 2b together with the nitrogen atom connecting them,

[0124] [ka] [In the formula, Each R ab is independently halo, hydroxyl, alkyl, haloalkyl, or alkoxy; X 2 O, NR x1 or CR x2 R x3 and R x1 , R x2 and R x3 are each independently H, halo, alkyl, preferably lower alkyl, or alkoxy; z is an integer having a value between 0 and 2. Form an optionally substituted heterocycle selected from:

[0125] In still other embodiments of the disclosed compounds, R 2 (CH2) n NR 2a R 2b where R 2a and R 2btogether with the nitrogen atom which connects them together to form an optionally substituted 3- to 6-membered heterocycle, and n is an integer having a value of 1 or 2. In some such embodiments, R 2a and R 2b together with the nitrogen atom connecting them,

[0126] [ka] [In the formula, Each R ab is independently halo, hydroxyl, alkyl, haloalkyl, or alkoxy; X 2 O, NR x1 or CR x2 R x3 and R x1 , R x2 and R x3 are each independently H, halo, alkyl, preferably lower alkyl, or alkoxy; z is an integer having a value between 0 and 2. Form an optionally substituted heterocycle selected from:

[0127] For example, an optionally substituted heterocycle is

[0128] [ka] You can choose from.

[0129] In some embodiments of the disclosed compounds, R 1 and R 2 together with the carbon atoms that connect them together form the structure

[0130] [ka] [In the formula, X 3 is NR Y1a or CRY1b R Y1c and X 4 is O or CR Y2a R Y2b and R Y1a is H, alkyl, -C(O)R Y1aa or -S(O)alkyl; R Y1aa is alkyl or alkoxy; R Y1b , R Y1c , R Y2a and R Y2b are each independently H or alkyl, preferably lower alkyl. It forms a heterocycle having the following structure:

[0131] In some such embodiments, the heterocycle is

[0132] [ka] is selected from.

[0133] In some embodiments of the disclosed compounds, X 1 is O or NR X1 and R X1 is H or alkyl, preferably lower alkyl; R 1 is methyl, and R 2 is optionally substituted alkyl, optionally substituted hydroxyalkyl, or (CR 2c 2) n NR 2a R 2b or R 1 and R 2 together with the carbon atom(s) connecting them together form a 5- or 6-membered heterocyclic ring having one N atom, which N atom is optionally substituted with lower alkyl; R 2ais H, methyl or ethyl, R 2b is H, methyl or ethyl, Each R 2c is independently H or alkyl, preferably methyl; n is an integer having a value of 1 to 4.

[0134] In some embodiments of the disclosed compounds, X 1 is O or NR X1 and R X1 is H, methyl or ethyl, R 1 is methyl, R 2 (CR 2c 2) n NR 2a R 2b and R 2a is H, methyl or ethyl, R 2b is H, methyl or ethyl, Each R 2c is independently H or alkyl, preferably methyl; n is an integer having a value of 1 to 4.

[0135] In some embodiments, each R 2c is H. In other embodiments, at least one R 2c is alkyl, preferably methyl, and the remainder are H.

[0136] In certain embodiments of the disclosed compounds, X 1 is O. In some such embodiments, (CR 2c 2) n NR 2a R 2b In the 2c is optionally alkyl, and the remainder are H. In other embodiments, X 1 is NR X1 In some such embodiments, (CR2c 2) n NR 2a R 2b In the 2c is optionally methyl, the remainder are H.

[0137] In certain embodiments of the disclosed compounds, R 2a and R 2b However, both cannot be H.

[0138] In some embodiments of the disclosed compounds, X 1 is O, R 1 is methyl, R 2 is optionally substituted hydroxyalkyl or optionally substituted C1-C4 alkyl-NHR 2a where R 2a is methyl or ethyl, or R 1 and R 2 together with the carbon atom(s) connecting them together form a 5- or 6-membered heterocycle having one N atom that is optionally substituted with lower alkyl.

[0139] In some embodiments of the disclosed compounds, X 1 is O or NR X1 and R X1 is H or alkyl, preferably lower alkyl; R 1 is methyl or ethyl, R 2 is lower alkyl, (CH2) n OH or (CR 2c 2) n NR 2a R 2b or R 1 and R 2together with the carbon atoms connecting them together form a 5- or 6-membered heterocyclic ring having one N atom substituted with -C(O)oxyalkyl; R 2a is H or lower alkyl optionally substituted with one or more halogens; R 2b is H or lower alkyl optionally substituted with one or more halogens, and R 2a and R 2b together with the N atom that connects them together form R ab z forming a 3-, 4- or 5-membered heterocyclic ring optionally substituted with R ab is halogen, hydroxyl, lower alkyl, haloalkyl, oxyalkyl; Each R 2c is independently H or alkyl, preferably methyl; z is an integer having a value of 0 to 2, n is an integer having a value of 2 to 4.

[0140] In some such embodiments, R 2a and R 2b However, both cannot be H.

[0141] In some embodiments of the disclosed compounds, X 1 is O or NR X1 and R X1 is H or alkyl, preferably lower alkyl; R 1 is methyl, R 2 is C1-C2 alkyl or (CH2) n NR 2a R 2b or R 1 and R 2together with the carbon atom(s) connecting them together form a 5- or 6-membered heterocyclic ring having one N atom optionally substituted with -C(O)alkyl; R 2a is unsubstituted lower alkyl; R 2b is unsubstituted lower alkyl, n is an integer having a value of 2 to 4.

[0142] In some embodiments of the disclosed compounds, X 1 is O or NR X1 and R X1 is H or alkyl, preferably lower alkyl; R 1 is methyl, R 2 is C1-C2 alkyl or (CR 2c 2) n NR 2a R 2b and R 2a is unsubstituted lower alkyl; R 2b is unsubstituted lower alkyl; Each R 2c is independently H or alkyl, preferably methyl; n is an integer having a value of 2 to 4.

[0143] In some embodiments of the disclosed compounds, X 1 is O or NR X1 and R X1 is H or alkyl, preferably lower alkyl; R 1 is alkyl, preferably lower alkyl; R 2 is C1-C3 alkyl, C1-C3 alkenyl, optionally substituted hydroxyalkyl, optionally substituted alkoxyalkyl or (CR 2c 2) n NR2a R 2b or R 1 and R 2 are taken together with the carbon atoms connecting them to form a 5- or 6-membered heterocycle having one heteroatom selected from N and O, and are optionally substituted with lower alkyl, carbonyl, tert-butyloxycarbonyl, -C(O)oxyalkyl or -S(O)2alkyl; R 2a and R 2b are each independently H, alkyl, or -C(O)alkyl; R 2a and R 2b together with the N atom that connects them together to form a 3-6 membered heterocycle in which one C is optionally replaced by O, and the heterocycle is (R ab ) z and optionally substituted with Each R ab are independently halogen, hydroxyl or lower alkyl; Each R 2c is independently H or alkyl, preferably methyl; z is an integer having a value of 1 or 2; n is an integer having a value of 2 to 4.

[0144] In certain embodiments, the compound of formula (I)

[0145] [ka] TIFF0007611822000033.tif215150TIFF0007611822000034.tif209149TIFF0007611822000035.tif205149TIFF0007611822000036.tif250149TIFF0007611822000037.tif244149TIFF0007611822000038.tif188149, or a pharma- ceutically acceptable salt thereof.

[0146] In certain embodiments, the compound of formula (I)

[0147] [ka] TIFF0007611822000040.tif217150TIFF0007611822000041.tif209150TIFF0007611822000042.tif205150TIFF0007611822000043.tif251150TIFF0007611822000044.tif196150TIFF0007611822000045.tif98150, or a pharma- ceutically acceptable salt thereof.

[0148] In some such embodiments, the compound of formula (I)

[0149] [ka] TIFF0007611822000047.tif198151 or a pharma- ceutically acceptable salt thereof.

[0150] In a preferred embodiment, the compound of formula (I)

[0151] [ka] or a pharma- ceutically acceptable salt thereof.

[0152] In another preferred embodiment, the compound of formula (I)

[0153] [ka] or a pharma- ceutically acceptable salt thereof.

[0154] In yet another preferred embodiment, the compound of formula (I)

[0155] [ka] TIFF0007611822000051.tif101152 or a pharma- ceutically acceptable salt thereof.

[0156] In yet another preferred embodiment, the compound of formula (I)

[0157] [ka]

[0158] [ka] or a pharma- ceutically acceptable salt thereof.

[0159] In another preferred embodiment, the compound of formula (I)

[0160] [ka] TIFF0007611822000055.tif98151 or a pharma- ceutically acceptable salt thereof.

[0161] In yet another preferred embodiment, the compound of formula (I)

[0162] [ka] TIFF0007611822000057.tif251151 or a pharma- ceutically acceptable salt thereof.

[0163] In yet another preferred embodiment, the compound of formula (I)

[0164] [ka]

[0165] [ka] or a pharma- ceutically acceptable salt thereof.

[0166] In another preferred embodiment, the compound of formula (I)

[0167] [ka] or a pharma- ceutically acceptable salt thereof.

[0168] In some embodiments, the compound of formula (I)

[0169] [ka] or a pharma- ceutically acceptable salt thereof.

[0170] In some embodiments, the compound is a mesylate salt.

[0171] In certain embodiments, the present invention provides a compound represented by formula (IVa) or (IVb):

[0172] [ka] A compound having the structure of TIFF0007611822000063.tif57144, or a pharma- ceutically acceptable salt thereof. [In the formula, X', independently at each occurrence, is halo, preferably F; R X1' is independently at each occurrence H or lower alkyl; R 1' is C1-C3 alkyl, R 2' is hydroxyalkyl or (CR 2c' 2) n' NR 2a' R 2b' and R 2a'is H, lower alkyl, acyl, or haloalkyl; R 2b' is H, lower alkyl, acyl, or haloalkyl; R 2a' and R 2b' together with the N atom that connects them together form R ab' z' or forming a 4-, 5- or 6-membered heterocyclic ring optionally substituted with R 1' and R 2' together with the C atom(s) which connect them together form a 5- or 6-membered heterocycle which is optionally substituted by acyloxy, R ab' is, when present, independently at each occurrence, halo, hydroxy, lower alkyl, or alkoxy; Each R 2c' is independently H or alkyl, preferably methyl; n' is an integer having a value of 1 or 2; z' is an integer having a value of 0, 1, or 2. and wherein the compound has a CDK4 K i or a pharma- ceutically acceptable salt thereof.

[0173] In some embodiments, the compound of formula (IVa) or (IVb) has an average IC of 150 nM or less for the drug-sensitive cell lines of Table 2. 50 has.

[0174] In certain embodiments, the average IC of the compound of formula (IVa) or (IVb) against the drug-sensitive cell lines in Table 2 50 is the average IC of the same compound of formula (IVa) or (IVb) against the drug-resistant cell lines in Table 2 50 It is at least about five times more potent than

[0175] In certain embodiments, the compound of formula (IVa) or (IVb) has a P of about 0.07 or greater. app Has an AB score.

[0176] In certain embodiments, the compound of formula (IVa) or (IVb) has a half-life of about 25 minutes or greater.

[0177] In some embodiments, the compound of formula (IVa) or (IVb)

[0178] [ka] TIFF0007611822000065.tif49144 or a pharmaceutical salt thereof.

[0179] In some embodiments, the present invention provides a method for the preparation of a compound of formula (V):

[0180] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, R 3a and R 3b together with the nitrogen atom to which they are attached form an optionally substituted [3.3] spiro ring moiety, wherein the optionally substituted [3.3] spiro ring moiety optionally contains at least one additional heteroatom selected from O, S and SO2. With respect to the above, excluding the compound

[0181] [ka] isn't it.

[0182] In some embodiments, the compound has the formula (Va):

[0183] [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, R 3a' and R 3b'together with the nitrogen atom to which they are attached

[0184] [ka] forming a structure selected from Each R ab is independently halo, hydroxyl, alkyl, haloalkyl, or alkoxy; z is 0, 1 or 2. has.

[0185] In certain embodiments, R 3a' and R 3b' together with the nitrogen atom to which they are attached,

[0186] [ka] Forming Each R ab are independently haloes, z is 2.

[0187] In some embodiments, R 3a' and R 3b' together with the nitrogen atom to which they are attached,

[0188] [ka] where R ab is fluoro.

[0189] In certain embodiments, the compound

[0190] [ka] or a pharma- ceutically acceptable salt thereof.

[0191] Treatment Method The compounds of the present invention are inhibitors of CDK4 / 6 and may therefore be useful in treating diseases whose underlying pathology is mediated (at least in part) by CDK4 / 6, including cancer and other diseases in which there is a disorder of cell proliferation, apoptosis or differentiation.

[0192] Examples of cancers that may be treated with the compounds of the invention include, but are not limited to, carcinomas such as cancers of the bladder, breast, colon (e.g., colorectal carcinoma, e.g., colon adenocarcinoma and colon adenoma), kidney, epidermis, liver, lung (e.g., adenocarcinoma, small cell lung carcinoma and non-small cell lung carcinoma), esophagus, gallbladder, ovary, pancreas (e.g., exocrine pancreatic carcinoma), stomach, cervix, thyroid, nose, head and neck, prostate, and skin (e.g., squamous cell carcinoma). Other examples of cancers that may be treated with the compounds of the invention include hematopoietic tumors of the lymphoid lineage (e.g., leukemia, acute lymphocytic leukemia, mantle cell lymphoma, chronic lymphocytic leukemia, B cell lymphoma (e.g., diffuse large B cell lymphoma), T cell lymphoma, multiple myeloma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, and Burkitt's lymphoma); hematopoietic tumors of the myeloid lineage, such as acute and chronic myeloid leukemia, myelodysplastic syndrome, and promyelocytic leukemia. Other cancers include follicular thyroid carcinoma; tumors of mesenchymal origin, such as fibrosarcoma or rhabdomyosarcoma; tumors of the central or peripheral nervous system, such as astrocytoma, neuroblastoma, glioma, or neurilemmoma; melanoma; seminoma; teratocarcinoma; osteosarcoma; xeroderma pigmentosum; retinoblastoma; keratoctanthoma; follicular thyroid carcinoma; and Kaposi's sarcoma.

[0193] One group of cancers includes human breast cancer (e.g., primary breast tumors, node-negative breast cancer, invasive ductal adenocarcinoma of the breast, non-endometrioid breast cancer); and endometrial cancer. Another subset of cancers in which compounds with CDK4 / 6 inhibitory activity may be particularly therapeutically beneficial includes glioblastoma multiforme, T-cell ALL, sarcoma, familial melanoma, and melanoma. CDK4 / 6 inhibitors are also useful in the treatment of viral infections, such as herpes viruses, pox viruses, Epstein-Barr virus, Sindbis virus, adenovirus, HIV, HPV, HCV, and HCMV; prevention of AIDS development in HIV-infected individuals; chronic inflammatory diseases, such as systemic lupus erythematosus, autoimmune-mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease, and autoimmune diabetes; cardiovascular diseases, such as cardiac hypertrophy, restenosis, atherosclerosis; neurodegenerative disorders, such as Alzheimer's disease, AIDS-associated dementia, Parkinson's disease, amyotropic lateral sclerosis, retinitis pigmentosa, spinal muscular atrophy, and the like. atropy and cerebellar degeneration; glomerulonephritis; myelodysplastic syndromes, ischemic injury-related myocardial infarction, stroke and reperfusion injury, arrhythmias, atherosclerosis, toxin-induced or alcohol-related liver disease, hematological diseases such as chronic anemia and aplastic anemia; degenerative diseases of the musculoskeletal system such as osteoporosis and arthritis, aspirin-sensitive sinusitis, cystic fibrosis, multiple sclerosis, kidney disease, age-related macular degeneration, eye diseases including uveitis, and cancer pain.

[0194] Furthermore, the compounds of the present invention may be useful for treating tumors with amplification of CDK4 and CDK6 genes, as well as tumors that overexpress cyclin partners of cyclin-dependent kinases. In particular, the compounds of the present invention may be useful for treating RB+ve (retinoblastoma protein positive) tumors, including tumors with mutations in Ras, Raf, growth factor receptors, or overexpression of growth factor receptors. In addition, the compounds of the present invention may be useful for treating RB-ve tumors.

[0195] The compounds of the present invention can also be useful in treating tumors with genetic abnormalities that activate CDK4 / 6 kinase activity.These include, but are not limited to, cancers with D-cyclin translocation, such as mantle cell lymphoma and multiple myeloma, cancers with D-cyclin amplification, such as breast cancer and esophageal squamous cell carcinoma, cancers with CDK4 amplification, such as liposarcoma, cancers with CDK6 amplification or overexpression, such as T-cell lymphoma, and cancers with p16 inactivation, such as melanoma, non-small cell lung cancer and pancreatic cancer.The compounds of the present invention can be useful in treating, and can be considered for treating, cancers with genetic abnormalities in upstream regulators of D-cyclin, whose defects lead to increased D-cyclin abundance. These include, but are not limited to, acute myeloid leukemia with FLT3 activation, breast cancer with Her2 / neu overexpression, ER-dependent or triple-negative phenotype, colon cancer with activating mutations in the MAPK, PI3K or WNT pathways, melanoma with activating mutations in the MAPK pathway, non-small cell lung cancer with aberrant activation of the EGFR pathway, and pancreatic cancer with aberrant activation of the MAPK pathway including K-ras mutations.

[0196] The methods of treatment of the present invention include administering a compound of the present invention or a pharma- ceutically acceptable salt thereof to a subject in need thereof. Individual embodiments of the present invention include a method of treating any one of the above-mentioned disorders or diseases by administering an effective amount of a compound of the present invention or a pharma- ceutically acceptable salt thereof to a subject in need thereof.

[0197] The pharmaceutical composition or combination of the present invention may be a unit dosage of about 1-1000 mg of active ingredient, or about 1-500 mg, or about 1-250 mg, or about 1-150 mg, or about 0.5-100 mg, or about 1-50 mg of active ingredient for a subject of about 50-70 kg. The therapeutically effective dosage of the compounds, pharmaceutical compositions or combinations thereof depends on the species, weight, age and individual condition of the subject, the disorder or disease being treated or its severity. A physician, clinician or veterinarian of ordinary skill can readily determine the effective amount of each active ingredient required to prevent, treat or inhibit the progression of the disorder or disease. The dosage characteristics listed above are demonstrable in vitro and in vivo tests, advantageously using mammals, such as mice, rats, dogs, monkeys, or isolated organs, tissues and preparations thereof. The compounds of the present invention can be applied in vitro in the form of solutions, e.g. aqueous solutions, and in vivo, either enterally, parenterally, advantageously intravenously, e.g. as a suspension or in aqueous solution. The in vitro dose is approximately 10 ~-9 Molar ~ 10 -3 The therapeutically effective amount in vivo may range from about 0.1 to 500 mg / kg, about 1 to 100 mg / kg, or about 100 to 300 mg / kg, depending on the route of administration.

[0198] Certain embodiments of the present invention include a method of modulating CDK4 / 6 activity in a subject, comprising administering to the subject a compound of the present invention or a pharma- ceutically acceptable salt thereof. Additional embodiments of the present invention provide a method for the treatment of a CDK4 / 6 mediated disorder or disease in a subject in need of such treatment, comprising administering to the subject a compound of formula (I), (Ia), (Ib), (IVa), (IVb), (V) or (Va) or a pharma- ceutically acceptable salt thereof. Other embodiments of the present invention provide a method for the treatment of a CDK4 / 6 mediated disorder or disease in a subject in need of such treatment, comprising administering to the subject a compound of the present invention or a pharma- ceutically acceptable salt thereof, wherein the disorder or disease is selected from carcinomas with genetic abnormalities that activate CDK4 / 6 kinase activity. These include, but are not limited to, cancers with D-cyclin translocations, such as mantle cell lymphoma and multiple myeloma, cancers with D-cyclin amplification, such as breast cancer and esophageal squamous cell carcinoma, cancers with CDK4 amplification, such as liposarcoma, cancers with CDK6 amplification or overexpression, such as T-cell lymphoma, and cancers with p16 inactivation, such as melanoma, non-small cell lung cancer, and pancreatic cancer.

[0199] The present invention also provides the use of a compound of the present invention, or a pharma- ceutically acceptable salt thereof, for the treatment of a disorder or disease mediated by CDK4.

[0200] In some embodiments, the compound of the present invention or its pharma- ceutically acceptable salt is used to treat CDK4-mediated disorders or diseases in subjects, wherein the disorders or diseases are selected from carcinomas with genetic abnormalities that activate CDK4 / 6 kinase activity.These include, but are not limited to, cancers with D-cyclin translocations, such as mantle cell lymphoma and multiple myeloma, cancers with D-cyclin amplification, such as breast cancer and esophageal squamous cell carcinoma, cancers with CDK4 amplification, such as liposarcoma, cancers with CDK6 amplification or overexpression, such as T-cell lymphoma, and cancers with p16 inactivation, such as melanoma, non-small cell lung cancer and pancreatic cancer.

[0201] Yet another embodiment of the present invention provides a compound according to formula (I), (Ia), (Ib), (IVa), (IVb), (V) or (Va) or a pharma- ceutically acceptable salt thereof for use as a medicament.

[0202] Yet another embodiment of the present invention includes the use of a compound of formula (I), (Ia), (Ib), (IVa), (IVb), (V) or (Va) or its pharma- ceutical acceptable salt in the manufacture of a medicament for the treatment of a disorder or disease mediated by CDK4 / 6, wherein the disorder or disease is selected from carcinomas with genetic abnormalities that activate CDK4 / 6 kinase activity. These include, but are not limited to, cancers with D-cyclin translocations, such as mantle cell lymphoma and multiple myeloma, cancers with D-cyclin amplification, such as breast cancer and esophageal squamous cell carcinoma, cancers with CDK4 amplification, such as liposarcoma, cancers with CDK6 amplification or overexpression, such as T-cell lymphoma, and cancers with p16 inactivation, such as melanoma, non-small cell lung cancer and pancreatic cancer.

[0203] combination The compounds of the invention may be administered simultaneously with, or conjointly before or after, one or more other therapeutic agents. The compounds of the invention may be administered separately by the same or different route of administration as the other agents, or together in the same pharmaceutical composition as the other agents.

[0204] In some embodiments, the present invention provides a product comprising a compound of the present invention or a pharma- ceutically acceptable salt thereof and at least one other therapeutic agent as a combined preparation for simultaneous, separate or sequential use in therapy. In some such embodiments, the therapy is the treatment of a disease or condition mediated by CDK4 / 6 inhibition. The product provided as a combined preparation includes a composition comprising a compound of the present invention and another therapeutic agent together in the same pharmaceutical composition, or comprising a compound of the present invention and another therapeutic agent in separate forms, for example, in the form of a kit. In certain embodiments, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharma- ceutically acceptable salt thereof and another therapeutic agent. Optionally, the pharmaceutical composition may comprise a pharma- ceutically acceptable excipient as described above.

[0205] In some embodiments, the present invention provides a kit comprising two or more separate pharmaceutical compositions, at least one of which contains a compound of the present invention or a pharma- ceutically acceptable salt thereof. In some such embodiments, the kit comprises a means for separately holding the compositions, such as a container, a divided bottle, or a divided foil packet. An example of such a kit is a blister pack, such as those typically used for packaging tablets, capsules, and the like.

[0206] The kit of the present invention can be used to administer different dosage forms, e.g., oral and parenteral, to administer separate compositions at different dosing intervals, or to titrate separate compositions against each other. To aid in compliance, the kit of the present invention typically includes instructions for administration. In the combination therapy of the present invention, the compound of the present invention and the other therapeutic agent can be manufactured and / or formulated by the same or different manufacturers. Furthermore, the compound of the present invention and the other therapeutic agent can be combined into a combination therapy: (i) before releasing the combination product to the physician (e.g., in the case of a kit containing the compound of the present invention and the other therapeutic agent); (ii) by the physician himself (or under the physician's guidance) immediately before administration; (iii) by the patient himself, e.g., during sequential administration of the compound of the present invention and the other therapeutic agent. Thus, the present invention provides the use of the compound of the present invention or a pharma- ceutically acceptable salt thereof for treating a disease or condition mediated by the inhibition of CDK4 / 6, where the medicament is prepared for administration with another therapeutic agent. The present invention also provides the use of another therapeutic agent for treating a disease or condition mediated by the inhibition of CDK4 / 6, where the medicament is administered with the compound of the present invention. The present invention also provides a compound of the present invention or a pharma- ceutically acceptable salt thereof for use in a method for treating a disease or condition mediated by CDK4 / 6 inhibition, wherein the compound of the present invention or a pharma- ceutically acceptable salt thereof is prepared for administration together with another therapeutic agent. The present invention also provides another therapeutic agent for use in a method for treating a disease or condition mediated by CDK4 / 6 inhibition, wherein the other therapeutic agent is prepared for administration together with the compound of the present invention or a pharma- ceutically acceptable salt thereof. The present invention also provides a compound of the present invention or a pharma- ceutically acceptable salt thereof for use in a method for treating a disease or condition mediated by CDK4 / 6 inhibition, wherein the compound of the present invention or a pharma- ceutically acceptable salt thereof is administered together with another therapeutic agent. The present invention also provides another therapeutic agent for use in a method for treating a disease or condition mediated by CDK4 / 6 inhibition, wherein the other therapeutic agent is administered together with the compound of the present invention or a pharma- ceutically acceptable salt thereof.The present invention also provides the use of a compound of the present invention or a pharma- ceutically acceptable salt thereof for treating a disease or condition mediated by CDK4 / 6, wherein the patient has previously (e.g., within 24 hours) been treated with another therapeutic agent. The present invention also provides the use of a compound of the present invention or a pharma- ceutically acceptable salt thereof for treating a disease or condition mediated by CDK4 / 6, wherein the patient has previously (e.g., within 24 hours) been treated with another therapeutic agent.

[0207] In some embodiments, the additional therapeutic agent is selected from an anti-inflammatory agent, an anti-proliferative agent, a chemotherapeutic agent, an immunosuppressant, an anti-cancer agent, a cytotoxic agent, or a kinase inhibitor other than a compound of the invention or a salt thereof.Further examples of agents that may be administered in combination with the compounds of the invention include, but are not limited to, PTK inhibitors, cyclosporin A, CTLA4-lg, anti-iCAM-3, anti-IL-2 receptor, antibodies selected from anti-CD45RB, anti-CD2, anti-CD3, anti-CD4, anti-CD80, anti-CD86, and monoclonal antibody OKT3, agents that block the interaction of CD40 with gp39, fusion proteins composed of CD40 and gp39, inhibitors of NF-kappa B function, nonsteroidal anti-inflammatory drugs, steroids, gold compounds, antiproliferative agents, FK506, mycophenolate mofetil, cytotoxic agents, TNF-a inhibitors, anti-TNF antibodies or soluble TNF receptors, rapamycin, mTOR inhibitors, leflunimide, cyclooxygenase-2 inhibitors, paclitaxel, cisplatin. , carboplatin, doxorubicin, carminomycin, daunorubicin, aminopterin, methotrexate, methopterin, mitomycin C, ecteinascidin 743, porfiromycin, 5-fluorouracil, 6-mercaptopurine, gemcitabine, cytosine arabinoside, podophyllotoxin, etoposide, etoposide phosphate, teniposide, melphalan, vinbrain Examples of inhibitors include vincristine, vincristine, leurocidin, epothilone, vindesine, leurosine, B-Raf inhibitors, MEK inhibitors, PI3K inhibitors, HSP90 inhibitors, CDK1 inhibitors, CDK2 inhibitors, CDK5 inhibitors, CDK7 inhibitors, CDK8 inhibitors, CDK9 inhibitors, EGFR inhibitors, FGFR inhibitors, PDGFR inhibitors, Her2 neu inhibitors, FLT3 inhibitors, androgen, glucocorticoid and prosterone receptor antagonists, SO inhibitors, WNT inhibitors, Bel inhibitors, IAP inhibitors, cl inhibitors, MD2 inhibitors, p52 inhibitors, proteosome inhibitors (Velcade), or derivatives thereof.

[0208] Specific individual combinations that may provide particular treatment benefits include co-treatment of mantle cell lymphoma or pancreatic cancer patients with mTOR inhibitors, such as everolimus.

[0209] In some embodiments, the compounds of the present invention may also be used in combination with other agents, e.g., additional protein kinase inhibitors, which may or may not be compounds of the present invention, for the treatment of protein kinase-associated disorders in a subject. The term "combination" means either a fixed combination in one unit dosage form or a kit-of-parts for co-administration, where the compound of the present invention and the combination partner can be administered independently and simultaneously, or separately within a time interval that specifically allows the combination partners to exhibit a cooperative effect, e.g., a synergistic effect, or any combination thereof.

[0210] The compounds of the invention may be administered simultaneously or sequentially with anti-inflammatory agents, anti-proliferative agents, chemotherapeutic agents, immunosuppressants, anti-cancer agents, cytotoxic agents, or kinase inhibitors other than the compounds of formula I or pharma- ceutically acceptable salts thereof. Further examples of agents that may be administered in combination with the compounds of the invention include, but are not limited to, PTK inhibitors, cyclosporin A, CTLA4-lg, anti-ICAM-3, anti-IL-2 receptor, antibodies selected from anti-CD45RB, anti-CD2, anti-CD3, anti-CD4, anti-CD80, anti-CD86, and monoclonal antibody OKT3, agents that block the interaction of CD40 with gp39, fusion proteins composed of CD40 and gp39, inhibitors of NF-kappa B function, nonsteroidal anti-inflammatory drugs, steroids, gold compounds, anti-proliferative agents, FK506, mycophenolate mofetil, cytotoxic agents, TNF-ct inhibitors, anti-TNF antibodies, or Examples of such inhibitors include soluble TNF receptors, rapamycin, leflunomide, cyclooxygenase-2 inhibitors, paclitaxel, cisplatin, carboplatin, doxorubicin, carminomycin, daunorubicin, aminopterin, methotrexate, methopterin, mitomycin C, ecteinascidin 743, porfiromycin, 5-fluorouracil, 6-mercaptopurine, gemcitabine, cytosine arabinoside, podophyllotoxin, etoposide, etoposide phosphate, teniposide, melphalan, vinblastine, vincristine, leurocidin, epothilone, vindesine, leurocidin, or derivatives thereof.

[0211] The compound of the present invention and any additional drug can be formulated in separate dosage forms.Alternatively, in order to reduce the number of dosage forms administered to a patient, the compound of the present invention and any additional drug can be formulated together in any combination.For example, the compound of the inhibitor of the present invention can be formulated in one dosage form, and the additional drug can be formulated together in another dosage form.Any separate dosage form can be administered at the same time or at different times.

[0212] Alternatively, the compositions of the present invention may contain additional agents as described herein. Each component may be present in an individual composition, a combination composition, or in a single composition.

[0213] Pharmaceutical Compositions and Their Administration The compositions and methods disclosed herein can be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal, such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or compound is preferably administered as a pharmaceutical composition, such as a disclosed compound and a pharma- ceutically acceptable carrier.

[0214] Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions, such as water or physiologically buffered saline, or other solvents or vehicles, such as glycols, glycerol, oils, such as olive oil, or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are for administration to humans, particularly invasive routes of administration (i.e., routes that avoid transport or diffusion through epithelial barriers, such as injection or implantation), the aqueous solutions are pyrogen-free or substantially pyrogen-free. Excipients can be selected, for example, to provide delayed release of the drug or to selectively target one or more cells, tissues, or organs. The pharmaceutical compositions can be in unit dosage form, such as tablets, capsules (including sprinkle capsules and gelatin capsules), granules, lyophilisates for reconstitution, powders, liquids, syrups, suppositories, injections, and the like. The compositions can also be present in transdermal delivery systems, such as skin patches. The compositions can also be present in liquids suitable for topical administration, such as ointments or creams.

[0215] A pharma- ceutically acceptable carrier can contain a physiologically acceptable agent that acts, for example, to stabilize, increase the solubility, or increase the absorption of a compound, for example, a compound of the present invention. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose, or dextran, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, or other stabilizers or excipients. The choice of a pharma- ceutical acceptable carrier that includes a physiologically acceptable agent depends, for example, on the route of administration of the composition. The pharmaceutical composition preparation can be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) can also be a liposome or other polymer matrix, in which, for example, the compound of the present invention can be incorporated. Liposomes, for example, liposomes containing phospholipids or other lipids, are non-toxic, physiologically acceptable, metabolizable carriers that are relatively simple to prepare and administer.

[0216] The phrase "pharmacologically acceptable" is used herein to refer to compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, and are commensurate with a reasonable benefit / risk ratio.

[0217] The phrase "pharmacologically acceptable carrier" as used herein means a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials that may serve as pharma- ceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and the like. soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances used in pharmaceutical preparations.

[0218] The pharmaceutical compositions (preparations) can be administered to a subject by any of several routes of administration, including, for example, orally (e.g., drenches, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue in aqueous or non-aqueous solutions or suspensions); absorption through the oral mucosa (e.g., sublingually); anally, rectally, or vaginally (e.g., as pessaries, creams, or foams); parenterally (e.g., as sterile solutions or suspensions, including intramuscularly, intravenously, subcutaneously, or intrathecally); nasally; intraperitoneally; subcutaneously; transdermally (e.g., as a patch applied to the skin); and topically (e.g., as a cream, ointment, or spray applied to the skin, or as eye drops). The compounds can also be formulated for inhalation. In certain embodiments, the compounds may simply be dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, and the patents cited therein.

[0219] The formulations can be conveniently presented in unit dosage form and can be prepared by any method known in the art of pharmacy. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form is generally the amount of compound that produces a therapeutic effect. Generally, this amount will be in the range of about 1% to about 99%, preferably about 5% to about 70%, most preferably about 10% to about 30% of the active ingredient out of 100%.

[0220] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, e.g., a compound of the present invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0221] Formulations of the present invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using flavored bases, usually sucrose and acacia or tragacanth), lyophilisates, powders, granules, or solutions or suspensions in aqueous or non-aqueous liquids, or as oil-in-water or water-in-oil liquid emulsions, or as elixirs or syrups, or as pastilles (using inert bases, such as gelatin and glycerin, or sucrose and acacia), and / or as mouthwashes, each of which contains a predetermined amount of a compound of the present invention as an active ingredient. The composition or compound may also be administered as a bolus, electuary, or paste.

[0222] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), the active ingredient is mixed with one or more pharma- ceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol; (4) hydrates, such as glycerol; (5) hydrates, such as glycerol; (6) hydrates, such as glycerol; (7) hydrates, such as glycerol; (8) hydrates, such as glycerol; (9) hydrates, such as glycerol; (10) hydrates, such as glycerol; (11) hydrates, such as glycerol; (12) hydrates, such as glycerol; (13) hydrates, such as glycerol; (14) hydrates, such as glycerol; (15) hydrates, such as glycerol; (16) hydrates, such as glycerol; (17) hydrates, such as glycerol; (18) hydrates, such as glycerol; (19) hydrates, such as glycerol; (19) hydrates, such as glycerol; (20) hydrates, such as glycerol; (21) hydrates, such as glycerol; (22) hydrates, such as glycerol; (23) hydrates, such as glycerol; (24) hydrates, such as glycerol; (2 ) disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) solution retarders such as paraffin; (6) absorption enhancers such as quaternary ammonium compounds; (7) wetting agents such as cetyl alcohol and glycerol monostearate; (8) absorbents such as kaolin and bentonite clay; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof; (10) complexing agents such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions may also include buffering agents. Solid compositions of similar types may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar, and high molecular weight polyethylene glycols.

[0223] Tablets can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface active agents or dispersants. Molded tablets can be made by molding a mixture of the powdered compound moistened with an inert liquid diluent in a suitable machine.

[0224] Tablets and other solid dosage forms of pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may be optionally scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may also be formulated to provide slow or controlled release of the active ingredient therein, for example, using different proportions of hydroxypropylmethylcellulose, other polymer matrices, liposomes and / or microspheres to provide the desired release profile. They may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or any other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents, and may be of a composition that releases the active ingredient(s) only, or preferentially, in a certain part of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, where appropriate, with one or more of the above-mentioned excipients.

[0225] Liquid dosage forms useful for oral administration include pharma- ceutically acceptable emulsions, lyophilisates for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs.In addition to active ingredients, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, cyclodextrin and its derivatives, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed, peanut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof.

[0226] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0227] Suspensions may contain, in addition to the active compound, suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.

[0228] Formulations of pharmaceutical compositions for rectal, vaginal or urethral administration may be presented as suppositories, which can be prepared by mixing one or more active compounds with one or more suitable non-irritating excipients or carriers including, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, which are solid at room temperature but liquid at body temperature and therefore will melt in the rectum or vaginal cavity and release the active compound.

[0229] Formulations of the pharmaceutical composition for administration to the mouth may be presented as a mouthwash, or an oral spray, or an oral ointment.

[0230] Alternatively or additionally, the compositions can be formulated for delivery via a catheter, stent, wire or other intraluminal device, which may be particularly useful for delivery to the bladder, urethra, ureter, rectum or intestine.

[0231] Formulations which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

[0232] Dosage forms for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharma- ceutically acceptable carrier, and with any preservatives, buffers, or propellants which may be required.

[0233] The ointments, pastes, creams and gels may contain, in addition to the active compound, excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and zinc oxide, or mixtures thereof.

[0234] Powders and sprays may contain, in addition to the active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays may additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0235] Transdermal patches have the additional advantage of providing controlled delivery of the compound of the present invention to the body.Such dosage forms can be made by dissolving or dispersing the active compound in a suitable medium.Absorption enhancers can also be used to increase the flux of the compound through the skin.The rate of such flux can be controlled by either providing a rate-controlling membrane or dispersing the compound in a polymer matrix or gel.

[0236] Ophthalmic formulations, eye ointments, powders, solutions, etc. are also contemplated within the scope of the present invention.Exemplary ophthalmic formulations are described in U.S. Patent Application Publication Nos. 2005 / 0080056, 2005 / 0059744, 2005 / 0031697, and 2005 / 004074; and U.S. Patent No. 6,583,124, the contents of which are incorporated herein by reference.If desired, liquid ophthalmic formulations have similar properties to tears, aqueous humor, or vitreous humor, or are compatible with such fluids.

[0237] The phrases "parenteral administration" and "administered parenterally", as used herein, mean modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

[0238] Pharmaceutical compositions suitable for parenteral administration contain one or more active compounds in combination with one or more pharma- ceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which can be reconstituted into sterile injectable solutions or dispersions immediately prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the preparation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0239] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0240] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of microbial action can be ensured by including various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like, in the compositions. In addition, prolonged absorption of the injectable pharmaceutical form can be brought about by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0241] In some cases, it is desirable to delay the absorption of drugs from subcutaneous or intramuscular injections in order to prolong the effect of the drug.This can be achieved by using liquid suspensions of crystalline or amorphous materials with poor water solubility.In this case, the absorption rate of the drug depends on its dissolution rate, which may further depend on the crystal size and crystalline form.Alternatively, delayed absorption of parenterally administered drug forms can be achieved by dissolving or suspending the drug in an oil vehicle.

[0242] Injectable depot forms are prepared by forming microencapsulated matrices of the subject compounds in biodegradable polymers, such as polylactide-polyglycolide. The rate of drug release can be controlled depending on the ratio of drug to polymer and the nature of the particular polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0243] For use in the methods of the present invention, the active compound may be provided per se or as a pharmaceutical composition containing, for example, 0.1% to about 99.5% (more preferably, about 0.5% to about 90.0%) of the active ingredient in combination with a pharma- ceutically acceptable carrier.

[0244] Incorporation by Reference All publications and patents mentioned in this specification are incorporated herein by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

[0245] Equivalent While specific embodiments of the present invention have been described, the above specification is illustrative and not restrictive. Many variations of the present invention will become apparent to those skilled in the art upon review of this specification and the following claims. The full scope of the present invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations. EXAMPLES

[0246] Example Synthesis Protocol

[0247] [Table 1] TIFF0007611822000074.tif221142TIFF0007611822000075.tif224142TIFF0007611822000076.tif220142TIFF000 7611822000077.tif220142TIFF0007611822000078.tif220142TIFF0007611822000079.tif223142TIFF00076118220 00080.tif221142TIFF0007611822000081.tif229142TIFF0007611822000082.tif229142TIFF0007611822000083.t if233142TIFF0007611822000084.tif234145TIFF0007611822000085.tif226144TIFF0007611822000086.tif102155

[0248] [Example 1] Compounds A1, A2, A3, A5, A6, A7, A8, A9, A10, A11, A12, A13, A14, A15, A16, A17, A18, A19, A 20, Synthesis of A21, A22, A29, A45, A47, A49, A50, A52, A53, A54, A55, A56, A57, A58, A59, A60

[0249] [ka] To a cooled (0 °C) solution of allyl 4-benzoyl-3-oxomorpholine-2-carboxylate (17.5 g, 60.5 mmol) in DMF (60 mL) was added iodomethane (7.5 mL, 121 mmol) followed by cesium carbonate (29.6 g, 90.8 mmol). The reaction was stirred for 2 h, at which point saturated ammonium chloride (aqueous, 300 mL) and water (200 mL) were added. The mixture was extracted with Et2O (3 x 300 mL) and the combined organic layers were dried over MgSO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of EtOAc in hexane = 0-50%) to afford allyl 4-benzoyl-2-methyl-3-oxomorpholine-2-carboxylate (14.6 g, 80% yield) as a white solid. MS: [M+H] + m / z 304.1.

[0250] [ka] To a Schlenk flask equipped with a stir bar was added Pd2(dba)3 (440 mg, 0.48 mmol), (S)-(p-CF3)3-t-BuPHOX (710 mg, 1.2 mmol) and MTBE (100 mL). The mixture was stirred for 30 min, after which allyl 4-benzoyl-2-methyl-3-oxomorpholine-2-carboxylate (14.6 g, 48 mmol) was added as a solution in MTBE (220 mL). The flask was sealed and heated to 50 °C for 2 days. Upon completion, the reaction was cooled to room temperature and vented to release the evolved CO2. The mixture was filtered through Celite, washed with EtOAc and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of EtOAc in hexanes = 0 to 20%) to give (S)-2-allyl-4-benzoyl-2-methylmorpholin-3-one (12.1 g, 98% yield, 98% ee) as a white solid. MS: [M+H] + m / z 260.1.

[0251] [ka] To a mixture of (S)-2-allyl-4-benzoyl-2-methylmorpholin-3-one (12.1 g, 47 mmol) in THF / MeOH (2:1, 24 mL) was added potassium carbonate (647 mg, 4.7 mmol). The flask was fitted with a reflux condenser and the reaction was heated to reflux for 3 h. Upon completion, the mixture was cooled, filtered through Celite, washed with CHCl, and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO (elution gradient of MeOH+2% NH in CHCl=0-20%) to afford (S)-2-allyl-2-methylmorpholin-3-one (6.45 g, 89% yield) as a colorless oil. MS: [M+H] + m / z 156.1.

[0252] [ka] To a cooled (0° C.) solution of (S)-2-allyl-2-methylmorpholin-3-one (6.45 g, 42 mmol) in THF (210 mL) was added lithium aluminum hydride (4.73 g, 125 mmol). The mixture was warmed to room temperature and heated to 60° C. for 3 h. Upon completion, the mixture was cooled to 0° C. and Et2O (300 mL) was added. Water (5.7 mL) was added slowly, followed by sodium hydroxide solution (1 M aqueous, 5.7 mL), and the mixture was warmed to room temperature and stirred for 15 min. The mixture was dried over MgSO4, filtered through Celite, and concentrated in vacuo to give (S)-2-allyl-2-methylmorpholine (5.29 g) as a colorless oil. The compound was used immediately without further purification. Mass [M + H] 142.1.

[0253] Compound A45

[0254] [ka] To a solution of (S)-2-allyl-2-methylmorpholine (5.29 g, 38 mmol) and 6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)nicotinaldehyde (12.75 g, 31 mmol) in DMSO (315 mL) was added sodium triacetoxyborohydride (13.23 g, 62 mmol) followed by acetic acid (18 mL, 312 mmol). The reaction was heated to 60° C. and stirred for 24 hours. Upon completion, the mixture was cooled to 0° C. and sodium hydroxide solution (1 M aq., 630 mL) was added followed by water (200 mL). The resulting heterogeneous mixture was stirred for 1 hour, then filtered to collect the solid, rinsed with cold water, and finally lyophilized to give (S)-N-(5-((2-allyl-2-methylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine (15.82 g) as a gray solid, which was used without further purification. MS: [M+H] + m / z 534.3.

[0255] Compound B1

[0256] [ka] To a solution of (S)-N-(5-((2-allyl-2-methylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine (15.82 g, 30 mmol) in THF / H2O (2:1, 750 mL) was added osmium tetroxide (4% in H2O, 9.4 mL, 1.48 mmol) followed by sodium periodate (19.02 g, 90 mmol). The reaction was sonicated for 2.5 h and then stirred for 4.5 h. Upon completion, saturated sodium thiosulfate (aq, 600 mL) was added followed by sodium hydroxide solution (1 M aq, 400 mL) and the mixture was extracted with CHCl3 (3 x 800 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo to give (S)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetaldehyde (15.94 g) as a brown solid. The compound was used without further purification. MS: [M+H] + m / z 536.3.

[0257] Compound A1

[0258] [ka] To a cooled (0° C.) solution of (S)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetaldehyde (15.94 g, 30 mmol) in CHCl3 / MeOH (2:1, 300 mL) was added dimethylamine (2 M in THF, 75 mL, 150 mmol). After stirring for 15 min, sodium triacetoxyborohydride (12.63 g, 60 mmol) was added and the reaction was allowed to warm to room temperature and stirred for 2 h. Upon completion, sodium hydroxide solution (0.5 M aq, 600 mL) was added and the mixture was extracted with CHCl3 (3×500 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of MeOH+2% NH3 in CHCl2=0-40%) to give (S)-N-(5-((2-(2-(dimethylamino)ethyl)-2-methylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine as a light orange oil. The oil was dissolved in water containing methanesulfonic acid (1.05 equiv.) and the solution was lyophilized to give 12.6 g of the mesylate salt as a tan solid. Mesylate salt spectrum 1H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 3.8 Hz, 1H), 8.30 (d, J = 1.3 Hz, 1H), 8.24 (dd, J = 8.5, 0.8 Hz, 1H), 8.22 - 8.18 (m, 1H), 7.75 - 7.64 (m, 2H), 4.85 (septet, J = 7.0 Hz, 1H), 3.59 (t, J = 4.8 Hz, 2H), 2.64 (s, 3H), 2.42 - 2.01 (m, 17H), 1.87 - 1.76 (m, 1H), 1.63 (d, J = 6.9 Hz, 6H), 1.53 (ddd, J = 13.2, 10.4, 5.9 Hz, 1H), 1.11 (s, 3H). MS: [M+H] + m / z 565.3.

[0259] Compound B2

[0260] [ka] (R)-2-(4-((6-((5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetaldehyde was prepared in the same manner as compound B1 (above) using (R)-(p-CF3)3-t-BuPHOX as the chiral catalyst. MS: [M+H] + m / z 536.3.

[0261] Compound A2

[0262] [ka] (S)—N-(5-((2-(2-(dimethylamino)ethyl)-2-methylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine and its mesylate salt from compound B1 in the same manner as described for compound A1 (above).

[0263] Compound A5

[0264] [ka] To a cooled (0° C.) solution of (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetaldehyde (90 mg, 0.17 mmol) in CHCl3 (2.1 mL) was added MgSO4 (120 mg) followed by methylamine (2M in MeOH, 0.84 mL, 1.7 mmol). After warming to room temperature and stirring for 20 h, sodium borohydride (13 mg, 0.34 mmol) was added and the reaction was stirred for 4 h. Upon completion, water (6 mL) and brine (2 mL) were added and the mixture was extracted with CHCl3 (3×4 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of MeOH+2% NH3 in CHCl2=0-40%) to give (R)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5-((2-methyl-2-(2-(methylamino)ethyl)morpholino)methyl)pyridin-2-yl)pyrimidin-2-amine as a light orange oil. The oil was dissolved in water containing methanesulfonic acid (1.05 equiv.) and the solution was lyophilized to give 28.3 mg of the mesylate salt as a tan solid. Mesylate salt spectrum 1H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 3.8 Hz, 1H), 8.29 (d, J = 1.3 Hz, 1H), 8.26 - 8.20 (m, 2H), 7.72 - 7.67 (m, 2H), 4.85 (septet, J = 6.8 Hz, 1H), 3.61 (t, J = 4.9 Hz, 2H), 2.64 (s, 3H), 2.60 - 2.53 (m, 4H), 2.37 - 2.27 (m, 9H), 2.20 (d, J = 11.1 Hz, 1H), 2.11 (d, J = 11.1Hz, 1H), 2.05 - 1.91 (m, 1H), 1.63 (d, J = 7.0 Hz, 6H), 1.58 - 1.45 (m, 1H), 1.12 (s, 3H). MS: [M+H] + m / z 551.3.

[0265] Compound A22

[0266] [ka] (S)-5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5-((2-methyl-2-(2-(methylamino)ethyl)morpholino)methyl)pyridin-2-yl)pyrimidin-2-amine and its mesylate salt from compound B1 in the same manner as described for compound A5 (above).

[0267] Compound A49

[0268] [ka] To a cooled (0° C.) solution of (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetaldehyde (1.37 g, 2.13 mmol) in CHCl3 / MeOH (2:1, 30 mL) was added sodium borohydride (121 mg, 3.2 mmol) and the reaction was stirred for 2 h. Upon completion, aqueous hydrochloric acid (1 M, 20 mL) was added and the mixture was stirred for a further 15 min. Sodium hydroxide solution (0.5 M, 200 mL) was added and the mixture was extracted with CHCl3 (3×150 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on C18 reverse-phase SiO2 (elution gradient of MeCN in HO+0.1% AcOH=5-50%) to give (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)ethan-1-ol as a light orange oil. The oil was dissolved in water containing methanesulfonic acid (1.05 equiv.) and the solution was lyophilized to give 400 mg of the mesylate salt as a tan solid. Mesylate salt spectrum 1 H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 2.8 Hz, 1H), 8.33 - 8.17 (m, 3H), 7.75 - 7.64 (m, 2H), 4.84 (p, J = 7.0 Hz, 1H), 3.65 - 3.55 (m, 2H), 3.49 - 3.39 (m, 2H), 2.65 (s, 3H), 2.39 - 2.25 (m, 6H), 2.24 - 2.07 (m, 2H), 1.97 - 1.83 (m, 1H), 1.69 - 1.52 (m, 8H), 1.14 (s, 3H). MS: [M+H] + m / z 538.2.

[0269] Compound A50

[0270] [ka] (S)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)ethan-1-ol and its mesylate salt from compound B1 in the same manner as described for compound A49 (above).

[0271] Compound A19

[0272] [ka] To a cooled (0° C.) solution of (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetaldehyde (30 mg, 0.056 mmol) in CHCl3 / MeOH (2:1, 1 mL) was added morpholine (50 μL, 0.56 mmol). After stirring for 15 min, sodium triacetoxyborohydride (24 mg, 0.11 mmol) was added and the reaction was allowed to warm to room temperature and stirred for 2 h. Upon completion, sodium hydroxide solution (0.5 M aq, 3 mL) was added and the mixture was extracted with CHCl3 (3×2 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of MeOH+2% NH3 in CHCl2=0-40%) to give (R)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5-((2-methyl-2-(2-morpholinoethyl)morpholino)methyl)pyridin-2-yl)pyrimidin-2-amine as a light orange oil. The oil was dissolved in water containing methanesulfonic acid (1.05 equiv.) and the solution was lyophilized to give 17.8 mg of the mesylate salt as a tan solid. Mesylate salt spectrum 1H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 3.8 Hz, 1H), 8.30 (d, J = 1.3 Hz, 1H), 8.28 - 8.19 (m, 2H), 7.75 - 7.65 (m, 2H), 4.85 (septet, J = 6.9 Hz, 1H), 3.60 (t, J = 4.8 Hz, 2H), 3.52 (t, J = 4.7 Hz, 4H), 2.65 (s, 3H), 2.44 - 2.17 (m, 13H), 2.10 (d, J = 11.1 Hz, 1H), 1.85 (q, J = 13.3, 10.7 Hz, 1H), 1.71 - 1.51 (m, 8H), 1.12 (s, 3H). MS: [M+H] + m / z 607.3.

[0273] Compound A7

[0274]

change

[0275] Compound A8

[0276]

Chem.

[0277] Compound A54

[0278]

Chem.

[0279] Compound A12

[0280] [ka] To a cooled (0 °C) solution of (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetaldehyde (30 mg, 0.056 mmol) in CHCl3 / MeOH (2:1, 1 mL) was added 3,3-difluoroazetidine·HCl (37 mg, 0.28 mmol). After stirring for 15 min, sodium triacetoxyborohydride (24 mg, 0.11 mmol) was added and the reaction was allowed to warm to room temperature and stirred for 4 h. Upon completion, sodium hydroxide solution (0.5 M aq, 3 mL) was added and the mixture was extracted with CHCl3 (3 × 2 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of MeOH+2% NH3 in CH2Cl2=0-40%) to give (R)-N-(5-((2-(2-(3,3-difluoroazetidin-1-yl)ethyl)-2-methylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine as a light orange oil. The oil was dissolved in water containing methanesulfonic acid (1.05 equiv.) and the solution was lyophilized to give 21.8 mg of the mesylate salt as a white solid. Mesylate salt spectrum 11H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 3.8 Hz, 1H), 8.31 (d, J = 1.4 Hz, 1H), 8.25 (d, J = 8.5 Hz, 1H), 8.22 (d, J = 2.2 Hz, 1H), 7.70 (m, 2H), 4.86 (p, J = 6.9 Hz, 1H), 3.59 (t, J = 5.0 Hz, 2H), 3.54 - 3.45 (m, 7H), 2.65 (s, 3H), 2.40 - 2.25 (m, 6H), 2.21 (d, J = 11.2 Hz, 1H), 2.10 (d, J = 11.2 Hz, 1H), 1.85 - 1.69 (m, 1H), 1.64 (d, J = 6.8 Hz, 6H), 1.45 - 1.35 (m, 1H), 1.12 (s, 3H). MS: [M+H] + m / z 613.3.

[0281] Compound A9

[0282]

Chem.

[0283] Compound A10

[0284]

Chem.

[0285] Compound A21

[0286]

change

[0287] Compound A13

[0288]

Chem.

[0289] Compound A15

[0290]

Chem.

[0291] Compound A14

[0292]

change

[0293] Compound A6

[0294]

change

[0295] Compound A29

[0296]

Chem.

[0297] Compound A20

[0298] [ka] To a cooled (0 °C) solution of allyl 4-benzoyl-3-oxomorpholine-2-carboxylate (2.0 g, 6.9 mmol) in DMF (7 mL) was added iodoethane (1.45 mL, 13.8 mmol) followed by cesium carbonate (4.5 g, 13.8 mmol). The reaction was stirred for 2 h, at which point saturated ammonium chloride (aqueous, 20 mL) and water (20 mL) were added. The mixture was extracted with Et2O (3 x 20 mL) and the combined organic layers were dried over Na2SO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of EtOAc in hexane = 0-50%) to afford allyl 4-benzoyl-2-ethyl-3-oxomorpholine-2-carboxylate (1.7 g, 78% yield) as a white solid. MS: [M+H] + m / z 317.1.

[0299] [ka] To a Schlenk flask equipped with a stir bar was added Pd(OAc)2 (6 mg, 0.027 mmol), (R)-(p-CF3)3-t-BuPHOX (80 mg, 0.134 mmol) and MTBE (40 mL). The mixture was stirred for 30 min, after which allyl 4-benzoyl-2-ethyl-3-oxomorpholine-2-carboxylate (1.7 g, 5.4 mmol) was added as a solution in MTBE (15 mL). The flask was sealed and heated to 55 °C for 4 days. Upon completion, the reaction was cooled to room temperature and vented to release the evolved CO2. The mixture was filtered through Celite, washed with EtOAc and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of EtOAc in hexanes = 0 to 20%) to give (R)-2-allyl-4-benzoyl-2-ethylmorpholin-3-one (1.3 g, 90% yield, 98% ee) as a white solid. MS: [M+H] + m / z 274.1.

[0300] [ka] To a cooled (0 °C) solution of (R)-2-allyl-4-benzoyl-2-ethylmorpholin-3-one (1.3 g, 4.8 mmol) in iPrOH (100 mL) was added lithium hydroxide (4 M in H2O, 1.8 mL, 7.1 mmol). The reaction was warmed to room temperature and stirred for 8 h. Upon completion, MgSO4 was added and the mixture was filtered through Celite, washed with CHCl2, and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of MeOH + 2% NH3 in CHCl2 = 0-30%) to give (R)-2-allyl-2-ethylmorpholin-3-one (367 mg, 45% yield) as a colorless oil. MS: [M+H] + m / z 170.1.

[0301] [ka] To a cooled (0° C.) solution of (R)-2-allyl-2-ethylmorpholin-3-one (367 mg, 2.2 mmol) in THF (11 mL) was added lithium aluminum hydride (250 mg, 6.5 mmol). The mixture was warmed to room temperature and heated to 60° C. for 2 h. Upon completion, the mixture was cooled to 0° C. and Et2O (11 mL) was added. Water (0.3 mL) was added slowly, followed by sodium hydroxide solution (1 M aqueous, 0.3 mL), and the mixture was warmed to room temperature and stirred for 15 min. The mixture was dried over MgSO4, filtered through Celite, and concentrated in vacuo to give (R)-2-allyl-2-ethylmorpholine (336 mg) as a colorless oil. The compound was used immediately without further purification. MS: [M+H] + m / z 156.2.

[0302] [ka] To a solution of (R)-2-allyl-2-ethylmorpholine (336 mg, 2.2 mmol) and 6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)nicotinaldehyde (739 mg, 1.8 mmol) in DMSO (18 mL) was added sodium triacetoxyborohydride (770 mg, 3.6 mmol) followed by acetic acid (1.1 mL, 18 mmol). The reaction was heated to 60° C. and stirred for 24 hours. Upon completion, the mixture was cooled to 0° C. and sodium hydroxide solution (1 M aq., 40 mL) was added followed by water (15 mL). The resulting heterogeneous mixture was stirred for 1 hour, then filtered to collect the solid, rinsed with cold water, and finally lyophilized to give (R)-N-(5-((2-allyl-2-ethylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine (862 mg) as a gray solid, which was used without further purification. MS: [M+H] + m / z 548.3.

[0303] [ka] To a solution of (R)-N-(5-((2-allyl-2-ethylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine (862 mg, 1.6 mmol) in THF / H2O (2:1, 39 mL) was added osmium tetroxide (4% in H2O, 0.5 mL, 0.08 mmol) followed by sodium periodate (1.00 g, 4.7 mmol). The reaction was sonicated for 2.5 h and then stirred for 4.5 h. Upon completion, saturated sodium thiosulfate (aq, 80 mL) was added followed by sodium hydroxide solution (1 M aq, 50 mL) and the mixture was extracted with CHCl3 (3 x 50 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo to give (R)-2-(2-ethyl-4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)morpholin-2-yl)acetaldehyde (815 mg) as a black solid. The compound was used without further purification. MS: [M+H] + m / z 550.3.

[0304] [ka] To a cooled (0° C.) solution of (R)-2-(2-ethyl-4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)morpholin-2-yl)acetaldehyde (70 mg, 0.13 mmol) in CHCl3 / MeOH (2:1, 2 mL) was added dimethylamine (2M in THF, 0.64 mL, 1.3 mmol). After stirring for 15 min, sodium triacetoxyborohydride (55 mg, 0.25 mmol) was added and the reaction was allowed to warm to room temperature and stirred for 2 h. Upon completion, sodium hydroxide solution (0.5 M aq, 6 mL) was added and the mixture was extracted with CHCl3 (3×3 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of MeOH+2% NH3 in CHCl2=0-40%) to give (R)-N-(5-((2-(2-(dimethylamino)ethyl)-2-ethylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine as a light orange oil. The oil was dissolved in water containing methanesulfonic acid (1.05 equiv.) and the solution was lyophilized to give 25.3 mg of the mesylate salt as a tan solid. Mesylate salt spectrum 1 H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 3.8 Hz, 1H), 8.30 (d, J = 1.3 Hz, 1H), 8.27 - 8.19 (m, 2H), 7.73 - 7.65 (m, 2H), 4.85 (septet, J = 6.9 Hz, 1H), 3.59 (m, 2H), 2.65 (s, 3H), 2.42 - 2.03 (m, 17H), 1.85 - 1.71 (m, 1H), 1.70 - 1.51 (m, 8H), 1.49 - 1.36 (m, 1H), 0.74 (t, J = 7.5 Hz, 3H). MS: [M+H] + m / z 579.3.

[0305] Compound A3

[0306] [ka] To a cooled (0° C.) solution of (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetaldehyde (303 mg, 0.57 mmol) in CHCl3 / MeOH (2:1, 7 mL) was added diethylamine (0.3 mL, 2.83 mmol). After stirring for 15 min, sodium triacetoxyborohydride (242 mg, 1.14 mmol) was added and the reaction was allowed to warm to room temperature and stirred for 2 h. Upon completion, sodium hydroxide solution (0.5 M aq, 14 mL) was added and the mixture was extracted with CHCl3 (3×10 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of MeOH+2% NH3 in CH2Cl2=0-40%) to give (S)-N-(5-((2-(2-(diethylamino)ethyl)-2-methylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine as a pale orange oil. The oil was dissolved in water containing methanesulfonic acid (1.05 equiv.) and the solution was lyophilized to give 230 mg of the mesylate salt as a white solid. Mesylate salt spectrum 11H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 3.9 Hz, 1H), 8.30 (d, J = 1.3 Hz, 1H), 8.24 (dd, J = 8.5, 0.8 Hz, 1H), 8.21 (dd, J = 2.4, 0.8 Hz, 1H), 7.74 - 7.65 (m, 2H), 4.86 (p, J = 6.8 Hz, 1H), 3.60 (t, J = 4.8 Hz, 2H), 2.71 - 2.51 (m, 4H), 2.47 - 2.26 (m, 12H), 2.23 (d, J = 11.0 Hz, 1H), 2.08 (d, J = 11.2 Hz, 1H), 1.87 - 1.72 (m, 1H), 1.63 (d, J = 6.9 Hz, 6H), 1.53 (ddd, J = 13.3, 10.3, 5.9 Hz, 1H), 1.11 (s, 3H), 0.90 (t, J = 7.1 Hz, 6H). MS: [M+H] + m / z 593.3.

[0307] Compound A16

[0308]

Chem.

[0309] Compound A11

[0310]

Chem.

[0311] Compound A17

[0312]

Chem.

[0313] Compound A18

[0314]

Chem.

[0315] Compounds A52 and A53

[0316]

Chem.

[0317] [ka] To a cooled (0° C.) solution of (S)-4-benzoyl-2-methyl-2-(2-oxopropyl)morpholin-3-one (43 mg, 0.16 mmol) in THF (1.6 mL) was added lithium aluminum hydride (59 mg, 1.6 mmol). The mixture was warmed to room temperature and heated to 50° C. for 22 h. Upon completion, the mixture was cooled to 0° C. and Et2O (2 mL) was added. Water (0.071 mL) was added slowly, followed by sodium hydroxide solution (1 M aq., 0.071 mL) and the mixture was warmed to room temperature and stirred for 15 min. The mixture was dried over MgSO4, filtered through Celite, and concentrated in vacuo to give 1-((S)-2-methylmorpholin-2-yl)propan-2-ol (25 mg) as a colorless oil and a mixture of diasteriomers in the alcohol. The compound was used immediately without further purification. MS: [M+H]+ m / z 160.1.

[0318] [ka] To a solution of 1-((S)-2-methylmorpholin-2-yl)propan-2-ol (25 mg, 0.16 mmol) and 6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)nicotinaldehyde (26 mg, 0.06 mmol) in DMSO (1.7 mL) was added sodium triacetoxyborohydride (41 mg, 0.19 mmol) followed by acetic acid (37 μL, 0.64 mmol). The reaction was heated to 60° C. and stirred for 16 h. Upon completion, the mixture was filtered and purified using preparative HPLC (elution gradient of MeCN in H20 + 0.25% TFA = 10-24%) to afford 1-((S)-4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)propan-2-ol (8 mg of major diastereomer, 6 mg of minor diastereomer) as a light brown solid. Spectrum of the free base Compound A52: Peak 1, minor diastereomer 1H NMR (400 MHz, cross-linked) δ 8.47 (d, J = 3.7 Hz, 1H), 8.44 (dd, J = 8.6, 0.8 Hz, 1H), 8.36 (s, 1H), 8.31 - 8.26 (m, 1H), 8.21 (d, J = 1.3 Hz, 1H), 7.82 (dd, J = 11.6, 1.4 Hz, 1H), 7.72 (dd, J = 8.6, 2.3 Hz, 1H), 4.76 (septet, J = 7.0 Hz, 1H), 4.25 (dtt, J = 12.8, 6.6, 3.2 Hz, 1H), 4.09 (s, 1H), 3.86 - 3.77 (m, 2H), 3.45 (q, J = 12.2 Hz, 2H), 2.72 (s, 3H), 2.55 (dt, J = 10.6, 3.9 Hz, 1H), 2.41 - 2.29 (m, 2H), 2.15 (d, J = 11.2 Hz, 1H), 1.90 (dd, J = 14.7, 10.5 Hz, 1H), 1.74 (d, J = 7.0 Hz, 6H), 1.40 (s, 3H), 1.32 (dd, J = 14.5, 1.4 Hz, 1H), 1.17 (d, J = 6.1 Hz, 3H). MS: [M+H] + m / z 552.3. Compound A53: ピーク1, main ジアステレオマー 1H NMR (400 MHz, cross-linked) δ 8.71 - 8.14 (m, 5H), 7.82 (dd, J = 11.5, 1.2 Hz, 1H), 7.73 (d, J = 8.6 Hz, 1H), 4.76 (septet, J = 6.9 Hz, 1H), 4.23 - 4.10 (m, 1H), 3.88 (ddd, J = 12.2, 9.4, 2.8 Hz, 1H), 3.75 (dt, J = 11.9, 3.4 Hz, 1H), 3.56 (d, J = 13.1 Hz, 1H), 3.39 (d, J = 13.2 Hz, 1H), 2.72 (s, 3H), 2.59 (d, J = 11.3 Hz, 1H), 2.44 (d, J = 11.4 Hz, 1H), 2.38 - 2.22 (m, 2H), 1.86 - 1.68 (m, 8H), 1.52 (dd, J = 14.9, 1.6 Hz, 1H), 1.35 (s, 3H), 1.16 (d, J = 6.2 Hz, 3H). MS: [M+H] + m / z 552.3.

[0319] Compound A47

[0320]

change

[0321] [ka] To a Schlenk flask equipped with a stir bar was added Pd(OAc)2 (3.7 mg, 0.017 mmol), (S)-(p-CF3)3-t-BuPHOX (25 mg, 0.041 mmol) and THF (7 mL). The mixture was stirred for 30 min, after which allyl 4-benzoyl-1-(4-methoxybenzyl)-2-methyl-3-oxopiperazine-2-carboxylate (141 mg, 0.33 mmol) was added as a solution in THF (4 mL). The flask was sealed and heated to 50° C. for 3 days. Upon completion, the reaction was cooled to room temperature and vented to release evolved CO2. The mixture was filtered through Celite, washed with EtOAc and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of EtOAc in hexanes = 0 to 30%) to give (S)-3-allyl-1-benzoyl-4-(4-methoxybenzyl)-3-methylpiperazin-2-one (78 mg, 62% yield, 90% ee) as a colorless oil. MS: [M+H] + m / z 379.2.

[0322] [ka] To a cooled (0° C.) solution of (S)-3-allyl-1-benzoyl-4-(4-methoxybenzyl)-3-methylpiperazin-2-one (78 mg, 0.21 mmol) in THF (2.2 mL) was added lithium aluminum hydride (78 mg, 2.1 mmol). The mixture was warmed to room temperature and heated to 50° C. for 18 h. Upon completion, the mixture was cooled to 0° C. and Et2O (3 mL) was added. Water (0.1 mL) was added slowly, followed by sodium hydroxide solution (1 M aqueous, 0.1 mL), and the mixture was warmed to room temperature and stirred for 15 min. The mixture was dried over MgSO4, filtered through Celite, and concentrated in vacuo to give (S)-2-allyl-4-benzyl-1-(4-methoxybenzyl)-2-methylpiperazine (18 mg) as a colorless oil. The compound was used immediately without further purification. MS: [M+H] + m / z 351.2.

[0323] [ka] To a solution of (S)-2-allyl-4-benzyl-1-(4-methoxybenzyl)-2-methylpiperazine (18 mg, 0.051 mmol) in EtOH / CHCl (4:1, 1.25 mL) was added 10% palladium on carbon (10 mg). The reaction vessel was placed in a pressure bomb and charged with 1000 psi of hydrogen gas. The system was heated to 60° C. for 2 days. Upon completion, the mixture was filtered through Celite, washed with CHCl / MeOH (1:1), and concentrated in vacuo to give (S)-2-methyl-2-propylpiperazine (7 mg) as a colorless oil. The compound was used immediately without further purification. MS: [M+H] + m / z 143.2.

[0324] [ka] To a solution of (S)-2-methyl-2-propylpiperazine (7 mg, 0.05 mmol) and 6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)nicotinaldehyde (10 mg, 0.025 mmol) in DMSO (0.5 mL) was added sodium triacetoxyborohydride (16 mg, 0.075 mmol) followed by acetic acid (14 μL, 0.25 mmol). The reaction was heated to 60° C. and stirred for 24 h. Upon completion, the mixture was filtered and purified using preparative HPLC (elution gradient of MeCN in H20+0.1% AcOH = 10-50%) to afford (S)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5-((3-methyl-3-propylpiperazin-1-yl)methyl)pyridin-2-yl)pyrimidin-2-amine (10 mg) as a white solid. Spectrum of the free base 1 H NMR (400 MHz, chloroform-d) δ 8.46 (d, J = 3.7 Hz, 1H), 8.42 (dd, J = 8.6, 0.8 Hz, 1H), 8.31 - 8.24 (m, 2H), 8.23 ​​(d, J = 1.3 Hz, 1H), 7.85 - 7.78 (m, 1H), 7.72 (dd, J = 8.6, 2.3 Hz, 1H), 4.76 (septet, J = 7.0 Hz, 1H), 3.45 (s, 2H), 2.95 (d, J = 5.4 Hz, 2H), 2.72 (s, 3H), 2.52 - 2.33 (m, 2H), 2.28 - 2.10 (m, 2H), 1.74 (d, J = 6.9 Hz, 6H), 1.68 - 1.49 (m, 1H), 1.47 - 1.19 (m, 4H), 1.13 (s, 3H), 0.93 (t, J = 6.9 Hz, 3H). MS: [M+H] + m / z 535.3.

[0325] Compound B3

[0326] [ka] To a cooled (0° C.) solution of (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetaldehyde (B2) (30 mg, 0.056 mmol) in tBuOH / MeCN (1:1, 1 mL) was added 2-methyl-2-butene (0.12 mL, 1.12 mmol) followed by the dropwise addition of a solution of sodium chlorite (50 mg, 0.56 mmol) and monosodium phosphate (87 mg, 0.56 mmol) in water (1 mL). The reaction was allowed to warm to room temperature and stirred for 5 h. Upon completion, water (1 mL) was added and the mixture was extracted with CHCl3 (3×2 mL). The organic layers were combined, dried over Na2SO4, and concentrated in vacuo to give (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetic acid (31 mg) as a brown solid. The compound was used without further purification. MS: [M+H] + m / z 552.3.

[0327] Compound A55

[0328] [ka] To a cooled (0° C.) solution of (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)acetic acid (31 mg, 0.056 mmol) in DMF (2.5 mL) was added N,N-diisopropylethylamine (50 μL, 0.28 mmol) followed by HATU (32 mg, 0.084 mmol). After stirring for 15 min, dimethylamine (2 M in THF, 84 μL, 0.17 mmol) was added and the reaction was stirred for 1.5 h. Upon completion, saturated sodium bicarbonate (aq, 5 mL) was added and the mixture was extracted with CHCl3 (3×2 mL). The organic layers were combined, dried over Na2SO4 and concentrated in vacuo. The crude compound was purified using flash chromatography on SiO2 (elution gradient of MeOH+2% NH3 in CHCl2=0-20%) to give (R)-2-(4-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-2-methylmorpholin-2-yl)-N,N-dimethylacetamide as a light orange oil. The oil was dissolved in water containing methanesulfonic acid (1.05 equiv.) and the solution was lyophilized to give 7.2 mg of the mesylate salt as a tan solid. Spectrum of the mesylate salt (signals broadened by amide rotamers) 1H NMR (400 MHz, DMSO-d6) δ 10.42 (s, 1H), 8.70 (d, J = 3.6 Hz, 1H), 8.36 (s, 1H), 8.29 (d, J = 8.8 Hz, 1H), 8.23 ​​(s, 1H), 7.86 (d, J = 8.9 Hz, 1H), 7.66 (d, J = 12.0 Hz, 1H), 4.80 (septet, J = 6.9 Hz, 1H), 4.31 (d, J = 22.5 Hz, 2H), 3.85 - 3.74 (m, 2H), 3.20 - 3.04 (m, 2H) 2.96 - 2.89 (m, 3H), 2.77 - 2.69 (m, 3H), 2.63 - 2.52 (m, 5H), 2.23 (s, 3H), 1.57 (d, J = 6.8 Hz, 6H), 1.36 (s, 2H), 1.28 - 1.06 (m, 3H). MS: [M+H] + m / z 579.3.

[0329] Compound A56

[0330]

change

[0331] Compound A57

[0332]

Chem.

[0333] Compound A58

[0334]

Chem.

[0335] Compound A59

[0336]

Chem.

[0337] Compound A60

[0338]

Chem.

[0339] Spectrum of the mesylate salt (signals broadened by amide rotamers) 1H NMR (400 MHz, DMSO-d6) δ 10.39 (s, 1H), 8.69 (d, J = 3.7 Hz, 1H), 8.36 (s, 1H), 8.29 (d, J = 8.6 Hz, 1H), 8.22 (d, J = 1.3 Hz, 1H), 7.85 (d, J = 8.9 Hz, 1H), 7.65 (d, J = 11.9 Hz, 1H), 4.79 (p, J = 6.9 Hz, 1H), 4.41 - 4.20 (m, 2H), 3.87 - 3.72 (m, 2H), 3.69 - 3.55 (m, 2H), 3.23 - 3.08 (m, 4H), 3.04 - 2.76 (m, 2H), 2.59 (s, 3H), 2.52 - 2.46 (m, 2H), 2.23 (s, 3H), 1.86 - 1.74 (m, 2H), 1.74 - 1.63 (m, 2H), 1.57 (d, J = 6.9 Hz, 6H), 1.17 (s, 3H). MS: [M+H] + m / z 605.3.

[0340] [Example 2] Synthesis of Compounds A43 and A44

[0341] [ka] A dry round bottom flask was charged with 2,2-dimethylpiperazine (896 mg, 7.8 mmol) followed by dichloromethane (40 mL) and triethylamine (1.63 mL, 11.8 mmol). The reaction flask was cooled to -78°C and benzyl chloroformate was injected (1.23 mL, 8.6 mmol) and the reaction was allowed to warm to ambient temperature for 3 h. The solvent was removed in vacuo and the reaction was purified by silica gel chromatography (0-10% methanol / dichloromethane). The main fraction was collected to give, after removal of the solvent, benzyl 3,3-dimethylpiperazine-1-carboxylate (1.96 g, quantitative). MS: [M+H] + m / z 249.

[0342] [ka] Benzyl 3,3-dimethylpiperazine-1-carboxylate (3.77 g, 15.0 mmol) was dissolved in dichloromethane (75 mL) under nitrogen and triethylamine (6.2 mL, 45.0 mmol) was injected followed by di-tert-butyl dicarbonate (3.98 g, 18.2 mmol). After 2 h, conversion was incomplete by TLC analysis and a second portion of di-tert-butyl dicarbonate (2.00 g, 9.1 mmol) was added and the reaction was stirred for 16 h. The reaction was concentrated in vacuo and purified by silica gel chromatography (ethyl acetate / hexanes 0-100%) to give 4-benzyl 1-(tert-butyl) 2,2-dimethylpiperazine-1,4-dicarboxylate as a clear oil (4.95 g, 95%). MS: [M+H] + m / z 349.

[0343] [ka] To a flask containing 4-benzyl 1-(tert-butyl) 2,2-dimethylpiperazine-1,4-dicarboxylate (4.95 g, 14.2 mmol) was added ethyl acetate (100 mL) followed by palladium on carbon (10%, 376 mg). Hydrogen gas was bubbled through the reaction mixture for 10 minutes and stirring was continued under a hydrogen atmosphere for 5 hours. At this point, TLC analysis indicated the reaction was complete (10% dichloromethane / methanol). The reaction was filtered through Celite and the filtrate was concentrated in vacuo to give tert-butyl 2,2-dimethylpiperazine-1-carboxylate (3.01 g, 99%) as a slightly yellow oil. MS: [M+H] + m / z 215.

[0344] Compound A44

[0345] [ka] tert-Butyl 2,2-dimethylpiperazine-1-carboxylate (352 mg, 1.6 mmol) was dissolved in dimethylsulfoxide (16.5 mL) and acetic acid (0.45 mL). 6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)nicotinaldehyde was added (336 mg, 0.8 mmol) and the reaction was stirred for 30 minutes followed by the addition of sodium triacetoxyborohydride (694 mg, 3.3 mmol). The reaction was warmed to 60° C. and heated for 17 hours before being cooled to ambient temperature and poured into a mixture of chloroform and 1N sodium hydroxide (100 mL portions). The organic phase was separated, dried over sodium sulfate, filtered and concentrated to dryness. The crude product was dissolved in dichloromethane and trifluoroacetic acid (5 mL aliquots) and stirred for 2 h. The reaction was concentrated in vacuo and dissolved in dimethylsulfoxide. After filtration, the filtrate was purified by reverse phase HPLC (10-60% acetonitrile / water + 0.25% trifluoroacetic acid over 15 min). Active fractions were pooled and concentrated on a Genevac to give a white solid which was partitioned between ethyl acetate and saturated sodium bicarbonate to give, after concentration of the organic phase, N-(5-((3,3-dimethylpiperazin-1-yl)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine (241 mg, 58%) as a white solid. The mesylate salt was prepared by treating an acetonitrile solution of the compound with aqueous methanesulfonic acid, then freezing the solution and concentrating it on a lyophilizer to a white solid. 11H NMR (500 MHz, methanol-d4) δ 8.60 (d, J = 3.8 Hz, 1H), 8.40 (dd, J = 8.6, 0.8 Hz, 1H), 8.37 (d, J = 1.3 Hz, 1H), 8.28 (dd, J = 2.4, 0.8 Hz, 1H), 7.89 - 7.82 (m, 2H), 5.01 - 4.96 (m, 1H), 3.65 (s, 2H), 3.32 (t, J = 5.2 Hz, 2H), 2.76 (s, 5H), 2.52 (s, 2H), 1.78 (d, J = 6.9 Hz, 6H), 1.46 (s, 6H). MS: [M+H] + m / z 507.

[0346] Compound A43

[0347]

Chem.

[0348] [Example 3] Synthesis of Compounds A30, A31, A34, A37, A39 and A41 A solution of tert-butyl-8-oxo-6-oxa-2,9-diazaspiro[4.5]decane-2-carboxylate (450 mg) in methanol was purified by SFC. Separation returned tert-butyl (S)-8-oxo-6-oxa-2,9-diazaspiro[4.5]decane-2-carboxylate (160.5 mg) and tert-butyl (R)-8-oxo-6-oxa-2,9-diazaspiro[4.5]decane-2-carboxylate (209.3 mg) as white solids. The following reaction was carried out on both enantiomers, but the yields and exact procedure from the first peak are reported.

[0349] [ka] A round bottom flask was charged with tert-butyl (S)-8-oxo-6-oxa-2,9-diazaspiro[4.5]decane-2-carboxylate (160.5 mg, 0.63 mmol) and tetrahydrofuran was injected to give a clear solution (4 mL). Borane in tetrahydrofuran was injected under nitrogen atmosphere (1 M, 1.9 mL, 1.9 mmol) and the reaction was stirred at ambient temperature for 3 h. The solvent was removed under reduced pressure and the residue was dissolved in methanol (5 mL). Palladium on carbon (10%, 10 mg) was added to the resulting solution and the suspension was stirred for 1 h and filtered through Celite. The filter cake was washed with dichloromethane. Concentration in vacuo gave tert-butyl (S)-6-oxa-2,9-diazaspiro[4.5]decane-2-carboxylate (112 mg, 73%). MS: [M+H] + m / z 243.

[0350] Compound A41

[0351] [ka] A round bottom flask was charged with tert-butyl (S)-6-oxa-2,9-diazaspiro[4.5]decane-2-carboxylate (112 mg, 0.45 mmol) and 6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)nicotinaldehyde (226 mg, 0.55 mmol). Dimethylsulfoxide (10 mL) and acetic acid (1 mL) were charged under nitrogen and the reaction was warmed to 60° C. for 18 hours. The reaction was cooled and diluted with sodium hydroxide (1N aqueous solution, 20 mL) and water (100 mL). A cloudy white precipitate formed and was stirred for 1 hour then filtered. The white solid was washed several times with water (5 mL each) and dried on a lyophilizer to give tert-butyl (S)-9-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-6-oxa-2,9-diazaspiro[4.5]decane-2-carboxylate (265 mg, 90%). 1 H NMR (500 MHz, DMSO-d6) δ 8.71 (d, J = 3.7 Hz, 1H), 8.31 (s, 1H), 8.26 (d, J = 8.5 Hz, 1H), 8.21 (d, J = 2.2 Hz, 1H), 7.73 - 7.67 (m, 2H), 4.86 (p, J = 7.0 Hz, 1H), 3.74 - 3.58 (m, 4H), 3.32 - 3.11 (m, 4H), 2.66 (s, 3H), 2.50 - 2.22 (m, 4H), 1.99 (d, J = 11.8 Hz, 1H), 1.82 (s, 1H), 1.65 (dd, J = 7.0, 3.1 Hz, 6H), 1.38 (s, 9H). MS: [M+H] + m / z 635.

[0352] Compound A34

[0353] [ka] tert-Butyl (S)-9-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-6-oxa-2,9-diazaspiro[4.5]decane-2-carboxylate (248 mg, 0.46 mmol) was dissolved in chloroform (6 mL) and trifluoroacetic acid was added via pipette (3 mL). The resulting orange solution was stirred for 30 min and concentrated to give (R)-N-(5-((6-oxa-2,9-diazaspiro[4.5]decan-9-yl)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine as the trifluoroacetate salt, which was used without further purification (301 mg, quantitative). An aliquot of this sample was free based with aqueous sodium bicarbonate and dichloromethane and purified by preparative HPLC (10-35% acetonitrile / water + 0.25% acetic acid over 15 min). This gave an analytical sample (3.9 mg) which was used for analysis by NMR and in biochemical and cell-based assays. 1 H NMR (500 MHz, DMSO-d6) δ 8.70 (d, J = 3.7 Hz, 1H), 8.30 (d, J = 1.3 Hz, 1H), 8.28 - 8.18 (m, 2H), 7.75 - 7.64 (m, 2H), 4.86 (p, J = 6.9 Hz, 1H), 3.58 - 3.55 (m, 2H), 3.46 (m, 2H), 2.77 (d, J = 11.2 Hz, 2H), 2.66 - 2.60 (m, 5H), 2.42 - 2.33 (m, 2H), 2.28 (d, J = 12.8 Hz, 1H), 1.76 (s, 1H), 1.68 - 1.59 (m, 6H). MS: [M+H] + m / z 535.

[0354] Compound A39

[0355] [ka] (R)-N-(5-((6-oxa-2,9-diazaspiro[4.5]decan-9-yl)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine 2,2,2-trifluoroacetate (32 mg, 0.049 mmol) was dissolved in dimethylformamide (3 mL) and treated with formaldehyde (0.10 mL, 37% in water, 1.68 mmol) followed by sodium triacetoxyborohydride (32 mg, 0.15 mmol). The reaction was stirred for 3 h and filtered through Celite. The filtrate was directly purified by HPLC (10-35% acetonitrile / water + 0.25% acetic acid over 15 min). Active fractions were pooled and concentrated on a lyophilizer to give (R)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5-((2-methyl-6-oxa-2,9-diazaspiro[4.5]decan-9-yl)methyl)pyridin-2-yl)pyrimidin-2-amine (4.7 mg, 18%) as a white solid. 1 H NMR (500 MHz, DMSO-d6) δ 8.71 (d, J = 3.7 Hz, 1H), 8.31 (d, J = 1.2 Hz, 1H), 8.27 - 8.21 (m, 2H), 7.74 - 7.67 (m, 2H), 4.87 (p, J = 6.9 Hz, 1H), 3.81 - 3.48 (m, 4H), 2.70 - 2.65 (m, 5H), 2.50 - 2.27 (m, 5H), 2.17 (s, 3H), 1.74 (q, J = 7.0, 6.3 Hz, 1H), 1.65 (d, J = 6.8 Hz, 6H). MS: [M+H] + m / z 549.

[0356] Compound A37

[0357] [ka] (R)-N-(5-((6-oxa-2,9-diazaspiro[4.5]decan-9-yl)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine 2,2,2-trifluoroacetate (32 mg, 0.049 mmol) was dissolved in dimethylformamide (3 mL) and treated with triethylamine (0.05 mL, 0.36 mmol) and acetyl chloride (10 μL, 0.18 mmol). The reaction was stirred for 3 h and quenched with a few drops of methanol. After filtration through Celite, the filtrate was purified by reverse phase HPLC (10-60% acetonitrile / water + 0.25% acetic acid over 15 min). Active fractions were pooled and concentrated on a lyophilizer to give (S)-1-(9-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-6-oxa-2,9-diazaspiro[4.5]decan-2-yl)ethan-1-one (4.0 mg, 14%) as a white solid. 1 H NMR (500 MHz, DMSO-d6) δ 8.71 (d, J = 3.7 Hz, 1H), 8.33 - 8.28 (m, 1H), 8.28 - 8.20 (m, 2H), 7.76 - 7.65 (m, 2H), 4.86 (p, J = 6.9 Hz, MS: [M+H] + m / z 577.

[0358] Compound A31

[0359] [ka] (R)-N-(5-((6-oxa-2,9-diazaspiro[4.5]decan-9-yl)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine 2,2,2-trifluoroacetate (32 mg, 0.049 mmol) was dissolved in dimethylformamide (3 mL) and treated with triethylamine (0.05 mL, 0.36 mmol) and mesyl chloride (10 μL). The reaction was stirred for 3 h and quenched with a few drops of methanol. After filtration through Celite, the filtrate was purified by reverse phase HPLC (10-60% acetonitrile / water + 0.25% acetic acid over 15 min). Active fractions were pooled and concentrated on a lyophilizer to give (S)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-(5-((2-(methylsulfonyl)-6-oxa-2,9-diazaspiro[4.5]decan-9-yl)methyl)pyridin-2-yl)pyrimidin-2-amine (3.8 mg, 13%) as a white solid. 1 H NMR (500 MHz, DMSO-d6) δ 8.71 (d, J = 3.8 Hz, 1H), 8.31 (d, J = 1.2 Hz, 1H), 8.28 - 8.17 (m, 2H), 7.76 - 7.67 (m, 2H), 4.86 (p, J = 6.9 Hz, 1H), 3.69 (t, J = 4.8 Hz, 2H), 3.50 - 3.45 (m, 3H), 3.28 (dd, J = 8.8, 5.3 Hz, 2H), 3.23 - 3.14 (m, 1H), 2.85 (s, 3H), 2.66 (s, 3H), 2.50 - 2.46 (m, 2H), 2.31 (d, J = 11.4 Hz, 2H), 2.09 - 1.96 (m, 1H), 1.87 (dt, J = 13.1, 8.8 Hz, 1H), 1.65 (d, J = 6.8 Hz, 6H). MS: [M+H] + m / z 613.

[0360] Compound A30

[0361] [ka] (R)-N-(5-((6-oxa-2,9-diazaspiro[4.5]decan-9-yl)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine 2,2,2-trifluoroacetate (32 mg, 0.049 mmol) was dissolved in dimethylformamide (3 mL) and treated with triethylamine (0.05 mL, 0.36 mmol) and methyl chloroformate (10 μL). The reaction was stirred for 3 h and quenched with a few drops of methanol. After filtration through Celite, the filtrate was purified by reverse phase HPLC (10-60% acetonitrile / water + 0.25% acetic acid over 15 min). Active fractions were pooled and concentrated on a lyophilizer to give methyl (S)-9-((6-((5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)pyridin-3-yl)methyl)-6-oxa-2,9-diazaspiro[4.5]decane-2-carboxylate (5.0 mg, 18%) as a white solid. 1 H NMR (500 MHz, DMSO-d6) δ 8.71 (d, J = 3.7 Hz, 1H), 8.35 - 8.16 (m, 3H), 7.78 - 7.62 (m, 2H), 4.86 (p, J = 6.8 Hz, 1H), 3.73 - 3.60 (m, 2H), 3.57 (s, 3H), 3.48 (s, 2H), 3.37 - 3.11 (m, 4H), 2.66 (s, 3H), 2.49 - 2.22 (m, 4H), 2.11 - 1.77 (m, 2H), 1.65 (d, J = 6.9 Hz, 6H). MS: [M+H] + m / z 593.

[0362] [Example 4] Synthesis of Compounds A23 and A28

[0363] [ka] To a cooled solution of (S)-2-allyl-2-methylmorpholine (6.74 g, 47.8 mmol) in dichloromethane (120 mL, 0.4 M) under nitrogen, triethylamine (9.80 mL, 71.7 mmol) was added and the mixture was cooled in an ice / water bath. 4-Toluenesulfonyl chloride (10.03 g, 52.3 mmol) was added in portions and the reaction was stirred for 30 min and warmed to ambient temperature for 90 min. Excess reagent was quenched by the addition of water (50 mL) and saturated sodium bicarbonate (50 mL). The mixture was transferred to a separatory funnel. The organic phase was separated and the aqueous phase was washed two more times with dichloromethane (30 mL). The combined organic washes were concentrated onto silica gel and the residue was dry loaded onto a silica flash column (gradient hexane / dichloromethane + 2% ethyl acetate: 0-100%). The active fractions were pooled and concentrated to give (S)-2-allyl-2-methyl-4-tosylmorpholine (11.95 g, 85%) as an off-white solid. MS: [M+H] + m / z 296.

[0364] [ka] A flask containing (S)-2-allyl-2-methyl-4-tosylmorpholine (11.95 g, 40.48 mmol) was charged with tert-butyl alcohol, methanol, and water (125 mL:75 mL:2 mL) and purged with nitrogen. To the resulting solution was added 1,4-benzoquinone (6.56 g, 60.72 mmol) and PdCl2(CH3CN)2 (313 mg, 1.21 mmol). The reaction was gently warmed to 50 °C and cooled after 3 h. The resulting solution was quenched with 1N hydrochloric acid (15 mL) and after 30 min, diluted with ethyl acetate (300 mL). The aqueous phase was separated and the remaining organic portion was washed several times with sodium hydroxide solution (1N, 50 mL portions) and saturated sodium bicarbonate (50 mL). The organic portion was concentrated onto silica gel and dry loaded onto a silica flash column (gradient hexane / ethyl acetate: 0-100%). The active fractions were pooled and concentrated to give (S)-1-(2-methyl-4-tosylmorpholin-2-yl)propan-2-one (10.55 g, 85%) as an off-white solid. MS: [M+H] + m / z 312.

[0365] [ka] Palladium on carbon (10%, 4.11 g) was placed in a glass bottle attached to a bomb reactor and gently purged with nitrogen gas to cover the catalyst bed. Under nitrogen, tetrahydrofuran (50 mL) was added by pipette and the mixture was stirred vigorously. Dimethylamine in tetrahydrofuran (100 mL, 2 M) was mixed with acetic acid (5.77 mL, 100 mmol) and the resulting solution was transferred by pipette into the catalyst solution. Finally, (S)-1-(2-methyl-4-tosylmorpholin-2-yl)propan-2-one (8.23 g, 26.4 mmol) was dissolved in tetrahydrofuran (50 mL) and transferred into the catalyst solution. The bomb reactor was sealed under a nitrogen atmosphere and attached to a high pressure hydrogen tank. The reactor was pressurized to 1000 psi and released three times. The pressurized reaction was then sealed and warmed to 60° C. for 38 hours. The reaction was cooled and the excess hydrogen gas was released. The catalyst solution was filtered through Celite and the cake was washed with ethyl acetate, the palladium was not dried. CAUTION: Residual palladium catalyst may be pyrophoric and was kept wet with ethyl acetate, then water, before being transferred to a waste container. After filtration, the filtrate was diluted with ethyl acetate (300 mL) and transferred to a separatory funnel. The organic phase was washed with sodium hydroxide solution (2N, 200 mL) and saturated sodium bicarbonate. The combined organic washes were concentrated onto silica gel and the residue was dry loaded onto a silica flash column (gradient dichloromethane / methanol: 0-25%). Three fractions were collected, the first containing the starting material, followed by (S)-N,N-dimethyl-1-((R)-2-methyl-4-tosylmorpholin-2-yl)propan-2-amine (2.33 g, 26%), and finally the product (S)-N,N-dimethyl-1-((S)-2-methyl-4-tosylmorpholin-2-yl)propan-2-amine (3.25 g, 36%). MS: [M+H] + m / z 341.

[0366] [ka] A round bottom flask was charged with (S)-N,N-dimethyl-1-((S)-2-methyl-4-tosylmorpholin-2-yl)propan-2-amine (2.84 g, 8.35 mmol). Tetrahydrofuran was injected under nitrogen (42 mL) and the resulting solution was cooled to -78°C. Lithium aluminum hydride (2.14 g, 56.36 mmol) was added in one portion and the reaction was allowed to warm to ambient temperature. The reaction was warmed to 60°C for 50.5 hours. At this point, the reaction was cooled again in a dry ice / acetone bath and sodium hydroxide (1N, 2.5 mL) and water (2.5 mL) were added dropwise resulting in vigorous evolution of hydrogen gas. After warming for 30 minutes, magnesium sulfate was added (20.0 g) and the reaction was diluted with 1 volume of diethyl ether and stirred vigorously. After 30 minutes, the mixture was filtered through Celite and the filter cake was washed with dichloromethane. The filtrate was concentrated to give (S)-N,N-dimethyl-1-((S)-2-methylmorpholin-2-yl)propan-2-amine (1.51 g, 97% crude) as a pure yellow oil. MS: [M+H] + m / z 187.

[0367] Compound A23

[0368] [ka] (S)-N,N-Dimethyl-1-((S)-2-methylmorpholin-2-yl)propan-2-amine (1.51 g, 8.35 mmol) was transferred to a round bottom flask and dissolved in DMSO (80 mL) and acetic acid (4.58 mL, 79.5 mmol). 6-((5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-yl)amino)nicotinaldehyde (3.24 g, 7.95 mmol) was added to give a milky suspension. After 30 min, sodium triacetoxyborohydride (5.05 g, 23.85 mmol) was added and the reaction was warmed to 60° C. for 14.5 h. After cooling, the resulting clear yellow solution was poured into sodium hydroxide (0.5N, 1 L) and the aqueous phase was washed three times with chloroform (600 mL each). The combined organic washes were dried over sodium sulfate, filtered, and concentrated onto silica gel. The residue was dry loaded onto a silica flash column (gradient dichloromethane / methanol 3% 17N methanol / ammonia containing: 0-50%, followed by a strong phase of methanol + 4% ammonium hydroxide). Active fractions were pooled and concentrated to give N-(5-(((S)-2-((S)-2-(dimethylamino)propyl)-2-methylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine (3.30 g, 67%). The free base was analyzed by proton NMR and LC / MS, which showed the desired product. The entire portion was dissolved in acetonitrile and aqueous methanesulfonic acid (0.23 M, 26 mL) was added. The salt solution was frozen in a dry ice acetone bath and concentrated on a lyophilizer over several days. 1H NMR (500 MHz, chloroform-d) δ 8.43 (d, J = 3.7 Hz, 1H), 8.39 (dd, J = 8.5, 0.8 Hz, 1H), 8.25 (dd, J = 2.3, 0.8 Hz, 1H), 8.22 (s, 1H), 8.19 (d, J = 1.3 Hz, 1H), 7.79 (dd, J = 11.5, 1.3 Hz, 1H), 7.69 (dd, J = 8.6, 2.3 Hz, 1H), 4.74 (p, J = 7.0 Hz, 1H), 3.76 (t, J = 4.9 Hz, 2H), 3.42 (s, 2H), 2.90 - 2.62 (m, 4H), 2.47 - 2.39 (m, 1H), 2.36 (q, J = 5.3 Hz, 1H), 2.33 - 2.25 (m, 1H), 2.25 - 2.18 (m, 1H), 2.18 (s, 6H), 1.90 (dd, J = 14.1, 3.8 MS: [M+H] + m / z 579.

[0369] Compound A28

[0370] [ka] (S)-N,N-dimethyl-1-((R)-2-methyl-4-tosylmorpholin-2-yl)propan-2-amine was taken on in the final two-step sequence in the same manner to give N-(5-(((S)-2-((R)-2-(dimethylamino)propyl)-2-methylmorpholino)methyl)pyridin-2-yl)-5-fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)pyrimidin-2-amine methanesulfonate. 1H NMR (500 MHz, クロロホルム-d) δ 8.44 (d, J = 3.8 Hz, 1H), 8.40 (dd, J = 8.5, 0.8 Hz, 1H), 8.30 (s, 1H), 8.25 (dd, J = 2.3, 0.8 Hz, 1H), 8.19 (d, J = 1.3 Hz, 1H), 7.82 - 7.77 (m, 1H), 7.69 (dd, J = 8.6, 2.3 Hz, 1H), 4.73 (td, J = 14.0, 7.1 Hz, 1H), 3.71 (t, J = 4.9 Hz, 2H), 3.51 - 3.31 (m, 2H), 2.83 (q, J = 6.3, 5.7 Hz, 1H), 2.69 (s, 3H), 2.38 - 2.34 (m, 2H), 2.24 (d, J = 11.1 Hz, 1H), 2.19 (s, 6H), 1.71 (d, J = 7.0 Hz, 6H), 1.60 (dd, J = 14.2, 7.1 Hz, 1H), 1.25 (s, 3H), 1.02 (d, J = 6.6 Hz, 3H). MS: [M+H] + m / z 579.

[0371] [Example 5] Synthesis of compounds A61~A64 Compound A61

[0372]

change

[0373] Compound A62

[0374]

Chem.

[0375] Compound A63

[0376]

Chem.

[0377] Compound A64

[0378]

Chem.

[0379] Biological Experiments General method Cell line proliferation delay assay Cells were seeded at a density of 1,000-5,000 cells per well in 48-well tissue culture plates. After a 24-hour resting period, cells were treated with compounds at 10 μM, 2 μM, 0.4 μM, 0.08 μM, 0.016 μM, and 0.0032 μM. A group of cells was treated with the vehicle in which the compounds were prepared and served as a control. Cells were grown in the presence of compounds for 6 days and counted on days 0 and 6. All cell counts were performed using a Synentec Cellavista plate imager. Cells that did not receive compound were counted on day 1 and this count was used as the baseline for calculation of growth inhibition. Growth inhibition was calculated as the ratio of cell population doublings in the presence of compound versus the absence of compound. If treatment resulted in a net loss of cells from baseline, the percent mortality was defined as the reduction in cell numbers in treated wells compared to the counts in untreated wells on day 1 after seeding. The IC of each compound was calculated using the 100% CI 0.01–0.02%. 50Values ​​(see Table 3; reported in μM) were calculated by fitting curves to the data points from each dose-response assay using the Proc NLIN function in SAS for Windows version 9.2 (SAS Institute, Inc.).

[0380] Cdk4 and 6 enzyme inhibition assays K i For the determination assay, 200 μM stock solutions of the compounds were subjected to semi-logarithmic serial dilutions using 100% DMSO as the solvent. Ten separate concentrations were diluted from 6×10 in 100% DMSO. -9 M was prepared as the dilution end point. 100% DMSO was used as a control. 10 μL from each of the serial dilutions was aliquoted into separate wells of a 96-well plate and 90 μL of water was added to each of those wells. The plate was shaken thoroughly and 5 μL from each of the wells of the plate was transferred into a well of the assay plate. The final volume of the wells in the assay plate was 50 μL. All compounds were diluted at 2×10 -6 M~6×10 -11 Ten assay concentrations ranging from 0.01 to 0.01 M were tested. The final DMSO concentration in the wells in the assay plate was 1% in all cases. i are shown in Table 3 in nM.

[0381] A radioactive protein kinase assay (33PanQinase® Activity Assay) was used to measure the kinase activity of six protein kinases. All kinase assays were performed in 96-well FlashPlates™ from PerkinElmer (Boston, MA, USA). All protein kinase assays contained 70 mM HEPES-NaOH pH 7.5, 3 mM MgCl2, 3 mM MnCl2, 3 μM Na-orthovanadate, 1.2 mM DTT, 50 μg / mL PEG20000, ATP [γ-33P]-ATP (approximately 9×1005 cpm / well), protein kinase and substrate. The reaction cocktail was incubated at 30° C. for 60 min. The reaction was stopped with 50 μL of 2% (v / v) H3PO4, and the plate was aspirated and washed twice with 200 μL of 0.9% (w / v) NaCl. 33 Pi incorporation was determined using a microplate scintillation counter (Microbeta, Wallac).

[0382] Caco-2 assay (P app AB (P app A to B) The extent of bidirectional human intestinal permeability of compounds was estimated using a Caco-2 cell permeability assay. Caco-2 cells were seeded on polyethylene membranes in 96-well plates. Growth medium was refreshed every 4-5 days until the cells formed a confluent cell monolayer. HBSS containing 10 mM HEPES at pH 7.4 was used as the transport buffer. Compounds were tested in duplicate at 2 μM in both directions. Digoxin, nadolol and metoprolol were included as standards. Digoxin was tested in duplicate at 10 μM in both directions, while nadolol and metoprolol were tested in duplicate at 2 μM in the AB direction. Final DMSO concentrations were adjusted to less than 1% for all experiments. Plates were incubated for 2 hours in a CO2 incubator at 37°C with saturated humidity and 5% CO2. After incubation, all wells were mixed with acetonitrile containing the internal standard and the plates were centrifuged at 4,000 rpm for 10 minutes. 100 μL of supernatant was collected from each well and diluted with 100 μL of distilled water for LC / MS / MS analysis. The concentrations of test and control compounds in the starting, donor and receiver solutions were quantified by LC / MS / MS using the analyte / internal standard peak area ratios. The apparent permeability coefficient, P app (cm / s) was calculated using the following formula: P app =(dC r / dt) × V r / (A×C0) [In the formula, dC r / dt is the cumulative concentration of compound in the receiver chamber as a function of time (μM / s); V r is the solution volume in the receiver chamber (0.075 mL on the apical side and 0.25 mL on the basolateral side); A is the surface area for transport, which is 0.0804 cm for the area of ​​the monolayer. 2 and C0 is the initial concentration in the donor chamber (μM). app The scores for the compounds are shown in Table 3. The discharge ratio was calculated using the following formula: Emission ratio=P app (BA) / P app (AB) The percent recovery was calculated using the following formula: Recovery rate (%) = 100 × [(V r ×C r )+(V d ×C d )] / (V d ×C0) [where Vd is the volume in the donor chamber, which is 0.075 mL on the apical side and 0.25 mL on the basolateral side; C d and C r are the final concentrations of transport compound in the donor and receiver chambers, respectively].

[0383] CYP enzyme inhibition assays The inhibition of various CYP isoenzymes was measured for each compound using a CYP enzyme inhibition assay in human liver microsomes (HLM). Compounds and standard inhibitors were prepared in 100x working solutions. Substrate or PBS was added to the corresponding wells, followed by compound, solvent or positive control working solution to the corresponding wells. HLM was added to a pre-warmed (37°C) plate and mixed with the NADPH cofactor for 10 min at 37°C. The reaction was stopped by adding 400 μL of cold stop solution (200 ng / mL tolbutamide and 200 ng / mL labetalol in ACN). The plate was centrifuged at 4,000 rpm for 20 min and the supernatant was transferred to 100 μL of HPLC water followed by LC / MS / MS analysis. IC of CYP isoenzymes 50 was averaged for each compound and is shown in Table 3 in μM.

[0384] Determining the metabolic stability of compounds The metabolic stability of the compounds was determined in hepatocytes from mice and rats. Compound half-lives are shown in Table 3 in minutes. Compounds were diluted to 5 μM in Williams' E medium from a 10 mM stock solution. 10 μL of each compound was aliquoted into wells of a 96-well plate, and the reaction was initiated by aliquoting 40 μL of a 625,000 cell / mL suspension into each well. The plate was incubated at 37° C. with 5% CO2. At each corresponding time point, the reaction was stopped by quenching with ACN containing an internal standard (IS) at 1:3. The plate was shaken at 500 rpm for 10 minutes and then centrifuged at 3,220×g for 20 minutes. The supernatant was transferred to another 96-well plate containing the diluted solution. The supernatant was analyzed by LC / MS / MS. Compound half-lives were estimated using the following formula: Residual compounds (%)= Peak area ratio of test compound to internal standard at the endpoint / Peak area ratio of test compound to internal standard at starting point

[0385] hERG inhibition assay The effects of compounds on hERG potassium channel conductance were measured using the automated patch clamp technique QPatch HTX Compounds were prepared at 10 mM stocks. CHO cells stably expressing the hERG potassium channel were used for this assay. Cells were cultured in a humidified and air-controlled (5% CO2) incubator at 37°C. Compound stocks and the positive control amitriptyline were dissolved in 100% DMSO to make various solution concentrations. These solutions were further diluted in the extracellular solution to achieve the final concentrations for testing. The final DMSO concentration in the extracellular solution was 0.30%. hERG QPatch HTX The assay was carried out at room temperature. The following voltage command protocol was used: From a holding potential of −80 mV, the voltage was first stepped to −50 mV for 80 ms for leak subtraction and then to +20 mV for 4,800 ms to open hERG channels. The voltage was then stepped back to -50mV over 5,000ms, generating a "rebound" or tail current that was measured and collected for data analysis. Finally, the voltage was stepped back to a holding potential of -80 mV for 3,100 ms. For each experiment, three additions of 5 μL vehicle were applied, followed by 30 runs of the baseline period voltage command protocol. The addition of increasing doses of each compound was then repeated three times (5 μL of compound each time). Percentage of control values ​​were calculated for compounds by taking the ratio of the current response in the presence of compound to the peak current in the presence of vehicle control and multiplying by 100% (see Table 3).

[0386] Compound solubility assay The relative kinetic solubility of each compound was determined at both low (1.2) and neutral (7.4) pH. Compounds were prepared at 10 mM stocks. Kinetic solubility was determined by UV and calibrated using a three-point standard curve (1, 20 and 200 μM). Compounds were shaken in 50 mM phosphate buffer pH 7.4 at room temperature for 24 hours or in SGF buffer pH 1.2 at 37° C. for 24 hours. Compound solubilities are reported in μM in Table 3.

[0387] Kinome analysis Recombinant kinases were produced in and purified from either BL21 strain E. coli or HEK-293 cells. Kinases were then tagged with DNA for qPCR detection. Streptavidin-coated magnetic beads were treated with biotinylated small molecule ligands for 30 minutes at room temperature to generate affinity resins for kinase assays. Ligand-bound beads were blocked with excess biotin and washed with blocking buffer (SeaBlock with 1% BSA, 0.05% Tween20, and 1 mM DTT) to remove unbound ligand and reduce nonspecific phage binding. Binding reactions were assembled by combining kinases, ligand-bound affinity beads, and compounds in binding buffer (20% SeaBlock, 0.17x PBS, 0.05% Tween20, 6mM DTT). Compounds were prepared as 40x stocks in 100% DMSO and diluted directly into the assay. All reactions were performed in polypropylene 384-well plates in a final volume of 0.02 mL. The assay plate was incubated at room temperature for 1 hour with shaking, and the affinity beads were washed with wash buffer (1x PBS and 0.05% Tween 20). The beads were then resuspended in elution buffer (1x PBS, 0.05% Tween 20, 0.5 μM non-biotinylated affinity ligand) and incubated at room temperature for 30 minutes with shaking. Kinase concentrations in the eluates were measured by qPCR. Primary screen binding interaction results are reported as "% of control", where lower numbers indicate stronger affinity for the test compounds.

[0388] Rodent xenograft studies Xenograft models of human cancer cell lines were cultured at 1.0–3.0 × 10 7 Tumors were established in 6-week-old CD-1 athymic nude mice by subcutaneous injection of 100 x 100 cells. 3 Once tumor xenografts reached an average size of 10 mm, mice (n=8) were randomized into treatment groups. Tumor xenografts were measured three times a week using calipers and tumor volumes (mm 3) was determined by multiplying height x width x length. For ER+ studies, 17-β-estradiol 60-day release pellets were implanted subcutaneously in the left flank 7 days prior to tumor inoculation. Statistical differences between treatment arms at specific time points were determined using paired two-tailed Student's t-tests. Differences between groups were considered statistically significant at p<0.05. Compounds were formulated in 1% HEC in 25 mM phosphate buffer (pH=2) and administered by daily oral gavage (PO). Data were analyzed using StudyLog software from StudyDirector (San Francisco, CA).

[0389] Rodent pharmacokinetic and single-dose saturation studies For pharmacokinetic (PK) analysis of compounds, tumor-free 6-week-old CD-1 athymic nude mice were administered a single PO dose of compound followed by saphenous vein blood sampling at the following times post-dose: 15, 30, 60, 120, 240, 480, and 1,440 min. No mouse was bled more than twice during the 1,440 min period. Untreated samples were collected from vehicle control animals. For plasma preparation, whole blood was collected in EDTA-treated tubes. Cells were removed from the plasma by centrifugation at 1,000-2,000 × g for 10 min using a refrigerated centrifuge. The plasma fraction was removed and stored at -80 °C. To determine the concentration at which drug exposure saturates, mice were dosed as above with increasing concentrations of compound covering the log concentration range (100 mg / kg to 1,000 mg / kg). Three mice were used for each collection time point and dose. Plasma samples were analyzed by mass spectrometry (HPLC) to determine the amount of compound in the peripheral circulation (ng / ml). For this analysis, 20 μL of plasma sample was mixed with 2 volumes of ice-cold internal standard solution (ISS) and centrifuged at 6,100 g for 30 min. The ISS contained acetonitrile with 100 ng / mL of compound, 50 ng / mL of dextromethorphan and 50 ng / mL of imipramine. An aliquot of the supernatant was transferred to an autosampler plate and diluted with 2 volumes of 0.2% formic acid in water. Specific analyte concentrations were determined against a standard curve (10,000-5 ng / ml) and mean concentrations + / - standard deviation were calculated.

[0390] Rodent Maximum Tolerated Dose Study For the maximum tolerated dose (MTD) determination study, tumor-free CD-1 athymic nude mice were randomized into five treatment groups (five mice per group) and treated with either 500, 400, 300, 200 or 100 mg / kg of compound by daily PO. Mice were weighed daily and weight loss (%) was calculated relative to the individual mouse weight at the start of treatment. The study continued for 14 days or until a group mean weight loss of >10% was observed in the animals. The MTD was determined as the highest dose at which a mean weight loss of <10% was observed over 14 days of administration.

[0391] Assignment of sensitive and resistant cohorts and mean IC 50 Calculating values Human cancer cell lines were grouped as "resistant" or "sensitive" to CDK4 / 6 inhibition based on whether their growth was slowed by ribociclib, palbociclib, and abemaciclib (see Table 2). These sensitive and resistant cohorts were examined for their response to each compound, and IC 50 was calculated for each cell line using the same technique as above. Mean IC for sensitive and resistant cohorts 50 was calculated as the geometric mean of the groups. The "therapeutic window" for each compound was calculated based on the mean IC 50 The mean IC of the drug-sensitive group 50 It was calculated by dividing by

[0392] [Table 2]

[0393] [Example 6] Single-treatment PK to identify saturating dose Plasma was collected at six time points (30 min, 60 min, 2 h, 4 h, 8 h and 24 h). The AUC and dose for saturation of exposure were determined. Three mice were used per time point, and the same mice were bled twice. 30 minutes and 4 hours 60 minutes and 8 hours 2 hours and 24 hours 9 mice per concentration Five concentrations (1.78x dilution, 1000mg / kg~100)

[0394] [Example 7] Dose tapering 14-day MTD 4 new mice (nice) were dosed starting at 500mg / kg until >.10% BWL (body weight loss), then reduced to 400mg / kg, etc. The MTD of abemaciclib (250, 200, 150, 100 mg / kg) was also defined.

[0395] [Example 8] Activity-guided selection of inhibitors A subgenus of CDK4 / 6 inhibitors with desirable properties was identified using a combination of in vitro data. In particular, the above-mentioned assays (e.g., cell line proliferation delay assay, Cdk4 and 6 enzyme inhibition assay, Caco-2 assay (P app A-B), determination of compound metabolic stability, and assignment of sensitive and resistant cohorts and mean IC 50 The results from the calculation of the ΔE(t) value were used to select compounds having the structural and functional characteristics defined in the subgenus of formula (IVa) and formula (IVb). In particular, selected compounds were tested in sensitive and resistant cell lines as described above, and the log difference between sensitive and resistant cohorts for selected compounds is shown in FIG. In general, CDK4 K i correlated with the size of the therapeutic window (i.e., the difference in potency between sensitive and resistant cell line cohorts) and the K i A smaller α is associated with a larger therapeutic window. This correlation is consistent with the CDK4 K i The reaction ends when the concentration exceeds approximately 0.960 nM. One of skill in the art will readily recognize that results from additional assays (e.g., CYP enzyme inhibition assays, hERG inhibition assays, compound solubility assays, kinome analysis, rodent xenograft studies, rodent pharmacokinetic and single dose saturation studies, rodent maximum tolerated dose studies, and oral bioavailability assays) may be used to identify other subgenuses of CDK4 / 6 inhibitors, or to narrow the subgenus determined using other results, e.g., the subgenus of Formula IVa and IVb.

[0396] [Table 3] TIFF0007611822000164.tif225163TIFF0007611822000165.tif225105

[0397] [Table 4] TIFF0007611822000167.tif249119 Some embodiments are given below. Item 1 Formula (Ia) or Formula (Ib): [ka] or a pharma- ceutically acceptable salt thereof. [In the formula, X is independently in each occurrence halo; R X1 is H or alkyl, R 1 is alkyl, and R 2 is optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkenyl, optionally substituted hydroxyalkyl, optionally substituted aminoalkyl, or optionally substituted amidoalkyl; R 1 and R 2 together with the carbon atom(s) connecting them together form an optionally substituted 5- or 6-membered heterocycle. Section 2 Formula (II):

change

change

change

change

change

change

change

change

change

change

Chem.

Chem.

Chem.

Chem.

化

Chem.

change

change

change

change

change

change

change

change

change

change

change

change

change

change

change

change

Claims

1. Formula (Ia) or Formula (Ib): 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof. [In the formula, X is independently in each occurrence halo; R X1 is H or alkyl, R 1 is alkyl, R 2 (CH 2 ) n C(O)NR 2a R 2b or (CH 2 ) n NR 2a R 2b and R 2a and R 2b each independently represents H, alkyl, haloalkyl, alkenyl, (CR c R d ) m OR e or -C(O)alkyl; R c , R d and R e are each independently H or alkyl; n is an integer having a value of 1 or 2, and m is an integer having a value from 2 to 5.

2. Formula (II): 【Chemistry 2】 2. The compound of claim 1 having the structure:

3. Formula (IIa): 【Chemistry 3】 3. The compound of claim 2 having the structure: or a pharma- ceutically acceptable salt thereof.

4. Formula (IIb): 【Chemistry 4】 3. The compound of claim 2 having the structure: or a pharma- ceutically acceptable salt thereof.

5. R 1 C 1 ~C 4 5. The compound according to claim 1, wherein:

6. R 1 is methyl or ethyl.

7. R 1 5. The compound according to claim 1, wherein is methyl.

8. R 2 But -(CH 2 ) 2 N(H)(CH 3 ), -(CH 2 ) 2 N(H)(C(CH 3 ) 3 ), -(CH 2 ) 2 N(H)(C(O)CH 3 ), -(CH 2 ) 2 N(H)(CH 2 CH 2 F), -(CH 2 ) 2 N(CH 3 )(CH 2 CH 2 F), -(CH 2 ) 2 N(CH 3 ) 2 , or -(CH 2 ) 2 N(CH 2 CH 3 8. The compound according to claim 1 , wherein

9. Formula (Ia) or Formula (Ib): 【Chemistry 5】 or a pharma- ceutically acceptable salt thereof. [In the formula, X is independently in each occurrence halo; R X1 is H, methyl or ethyl, R 1 is methyl, R 2 (CR 2c 2 ) n NR 2a R 2b and R 2a is H, methyl or ethyl, R 2b is H, methyl or ethyl, Each R 2c is independently H or alkyl; and n is an integer having a value from 1 to 4.

10. At least one R 2c 10. The compound of claim 9, wherein is lower alkyl.

11. At least one R 2c is lower alkyl and the remainder are H.

12. At least one R 2c 10. The compound of claim 9, wherein is methyl.

13. At least one R 2c is methyl and the remainder are H.

14. R 2a and R 2b and n are not both H.

15. R 1 is methyl, R 2a is unsubstituted lower alkyl; R 2b is unsubstituted lower alkyl; 2. The compound of claim 1, wherein n is 2.

16. R 2a and R 2b are each independently H, alkyl, or —C(O)alkyl; 2. The compound of claim 1, wherein n is 2.

17. 【Chemical 6】 or a pharma- ceutically acceptable salt thereof.

18. The compound is a compound of formula (Ia) [In the formula, R 1 is C 1 ~C 3 is alkyl, R 2 (CR 2c 2 ) n NR 2a R 2b and R 2a is H, lower alkyl, —C(O)alkyl or haloalkyl; R 2b is H, lower alkyl, —C(O)alkyl or haloalkyl; R 2c is H] and the compound has a CDK4 K i 2. The compound of claim 1, having the formula:

19. The table below 【Table 1】 Average IC of 150 nM or less for drug-sensitive cell lines 50 20. The compound of claim 18, having the formula:

20. The table below 【Table 2】 The average IC of the compound against drug-sensitive cell lines 50 But the following table 【Table 3】 The average IC of the compound against drug-resistant cell lines 50 20. The compound of claim 18, which is at least 5-fold more potent than

21. P ≥ 0.07 app 21. The compound according to any one of claims 18 to 20, having an AB score.

22. 22. The compound of any one of claims 18 to 21, having a half-life of 25 minutes or more.

23. 【Chemical 7】 19. The compound of claim 18, selected from:

24. 【Chemical 8】 【change】 【change】 or a pharma- ceutically acceptable salt thereof.

25. 25. A pharmaceutical composition comprising a compound according to any one of claims 1 to 24 and a pharma- ceutically acceptable diluent or excipient.

26. 27. Use of a compound selected from any one of claims 1 to 24, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of cancer.

27. The use of claim 26, wherein the cancer is selected from carcinoma of the bladder, breast, colon, kidney, epidermis, liver, lung, esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, nose, head and neck, prostate, or skin; hematopoietic tumors of the lymphoid lineage; hematopoietic tumors of the myeloid lineage; tumors of mesenchymal origin; tumors of the central or peripheral nervous system; seminoma; teratocarcinoma; osteosarcoma; xeroderma pigmentosum; retinoblastoma; keratoacanthoma; thyroid follicular carcinoma; Kaposi's sarcoma; human breast cancer; endometrial carcinoma; glioblastoma multiforme; T-cell acute lymphoblastic leukemia (ALL); sarcoma; familial melanoma; and melanoma.

28. 25. Use of a compound selected from any one of claims 1 to 24, or a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament for sensitizing cancer cells and / or tumor cells to a chemotherapeutic agent or radiation.

29. 29. The use of claim 28, wherein the cancer and / or tumor cells have a D-cyclin translocation, a D-cyclin amplification, a CDK4 amplification, or a CDK6 amplification or overexpression, and / or the cancer has a genetic abnormality in an upstream regulator of a D-cyclin.

30. The use according to claim 28 or 29, wherein the cancer and / or tumor is selected from mantle cell lymphoma, multiple myeloma, breast cancer, esophageal squamous cell carcinoma, liposarcoma, non-small cell lung cancer, and pancreatic cancer, or the cancer is selected from acute myeloid leukemia with FLT3 activation, breast cancer with Her2 / neu overexpression, ER-dependent or triple-negative phenotype, colon cancer with activating mutations in the MAPK, PI3K or WNT pathways, melanoma with activating mutations in the MAPK pathway, non-small cell lung cancer with abnormal activation of the EGFR pathway, and pancreatic cancer with abnormal activation of the MAPK pathway including K-ras mutation.

31. 28. The use of claim 27, wherein the cancer is selected from colon adenocarcinoma and colon adenoma; adenocarcinoma, small cell and non-small cell lung carcinoma; exocrine pancreatic carcinoma; squamous cell carcinoma; leukemia, acute lymphocytic leukemia, mantle cell lymphoma, chronic lymphocytic leukemia, B cell lymphoma, diffuse large B cell lymphoma, T cell lymphoma, multiple myeloma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, and Burkitt's lymphoma; acute and chronic myeloid leukemia, myelodysplastic syndromes, promyelocytic leukemia, follicular thyroid carcinoma; fibrosarcoma or rhabdomyosarcoma; astrocytoma; neuroblastoma, glioma or schwannoma; and primary breast tumors, node-negative breast cancer, invasive ductal adenocarcinoma of the breast or non-endometrioid breast carcinoma.

Images