Novel pyrrolopyrimidine compounds as inhibitors of protein kinases

[0001] The present application takes advantage of Priority US Application No. 61 / 680,231, filed August 6, 2012, entitled "NOVEL EGFR MODULATORS AND USES THEREOF", US Provisional Application No. 61 / 814,147, filed April 19, 2013, entitled "NOVEL PYRROLOPYRIMIDINE COMPOUNDS. AS INHIBITORS OF PROTEIN KINASES ", US Patent Application No. 13 / 843,554, filed March 15, 2013, entitled" NOVEL EGFR MODULATORS AND USES THEREOF "and US Patent Application No. 13 / 917.514, filed on June 13, 2013, entitled" NOVEL EGFR MODULATORS AND USES thereof. " This application in some respects relates to US Provisional Application No. 61 / 586,718, filed January 13, 2012, entitled "Heterocyclic Compounds and Uses as Anticancer Agents" and to US Patent Application No. 13 / 740,182, filed January 12, 2013,

Field of Technology [0002] The field of the invention are pharmaceutical compounds, compositions and methods, in particular because they are associated with compositions and methods of treating proliferation disorders and other diseases associated with deregulation of kinases (such as, but not limited to, EGFR (including HER). , Alk, PDGFR, BLK, BMX / ETK, BTK, FLT3 (D835Y), ITK, JAK1, JAK2, JAK3, TEC and TXK) and / or corresponding pathways.

Background Fields [0003] Protein kinases are a group of enzymes that regulate a variety of important biological processes including cell growth, proliferation, survival, invasion and differentiation, organ formation, tissue repair and regeneration, etc. Protein kinases perform physiological functions by catalyzing protein phosphorylation and thus modulating cellular activity. Because protein kinases have a profound effect on cells, their activities are tightly regulated. The kinases are switched on or off by phosphorylation (sometimes by autophosphorylation), by binding activator proteins or inhibitor proteins, or small molecules, or by controlling their position in cells relative to their substrates. It is known that disturbances in kinase activity associated with genetic disorders or environmental factors are associated with many diseases.

[0004] The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is a member of the ErbB receptor family, a subfamily of four closely related tyrosine kinase receptors: EGFR (ErbB-1), HER2 / c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). EGFR is the cell surface receptor for members of the epidermal growth factor family (EGF family) of extracellular protein ligands. Mutations affecting EGFR expression or activity could lead to cancer. EGFR is reported to deregulate in the majority of solid tumor types, i.e. lung cancer, breast cancer and brain tumor. It is estimated that mutations, amplifications or incorrect regulation of EGFR or family members are involved in approximately 30% of all epithelial tumors. Therapeutic approaches have been developed based on the inhibition of EGFR by either an antibody-drug or a small-molecule inhibitor drug such as gefitinib and erlotinib. For non-small cell lung cancer, gefitinib and erlotinib showed benefits for approximately

10-40% of patients. However, the acquired resistance to gefitinib or erlotinib after a period of treatment becomes a serious clinical problem. Studies have confirmed that one of the main reasons for developing resistance is due to the presence of a new T790M mutation that is the guardian of the EGFR genome. Subsequently, inhibitors have been developed that can overcome this developed T790M and have been shown to be superior in clinical trials, e.g. BIBW2992. However, this EGFR inhibitor targeting T790M still has relative inhibitory activity against wild-type EGFR, which limits clinical application. It is necessary to further develop a more effective type of EGFR inhibitor that will target substantially the mutation and essentially no wild-type protein.

[0005] Other protein kinases that are useful for small molecule pharmaceuticals include lymphoid B tyrosine kinase (BLK), Janus 1 kinase (JAK1), bone marrow kinase X (BMX / ETK), Bruton tyrosine kinase (BTK), Janus 2 kinase (JAK2), Janus 3 kinase (JAK3), hepatic kinase expressed in hepatocellular carcinoma (TEC), resting lymphocyte kinase (TXK, also known as RLK), FMS-like tyrosine kinase 3 (FLT3) and FLT3 (D835Y) .

Summary [0006] The present invention is directed to a certain pyrrolopyrimidine derivative and pharmaceutical compositions and to these compounds and compositions for use in the treatment of proliferative disorders and others.

[0007] The present disclosure provides a compound of Formula (VIII):

<img file="PL2880035T3_D0001.tif" />

wherein

X¹ is O, NH, S, CH2, or CF2;

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro;

n is a number from zero to 4;

R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and -NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C1-6 alkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

Q is CR⁹ or N;

where R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl;

R¹³ is selected from hydrogen, C1-6 alkyl, C1-6 acyl, SO2-C1-6alkyl, C3-7 cycloalkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or Hello; and -NR¹⁸R¹⁹ means (a)

<img file="PL2880035T3_D0002.tif" />

where R¹⁰ is selected from hydrogen and C1-6 alkyl;

R¹⁵ is unsubstituted methyl, or is C2-4alkyl unsubstituted or substituted hydroxy, methoxy, or halo; and m is 1 or 2;

or means (b), where R¹⁹ and R⁹ taken together form a 5- or 6-membered heteroaryl ring optionally substituted with C 1-6 alkyl which is unsubstituted or substituted with amino, hydroxyl, or halo; and R¹⁸ is hydrogen or C 1-6 alkyl, or is absent to satisfy the valence of the heteroaryl ring;

provided that none of R⁶ or R⁷ does not mean methoxy when NR¹⁸R¹⁹ means r1 ° -N^WITH ?

or a pharmaceutically acceptable salt thereof.

[0008] The present disclosure provides a compound of Formula (Ia) and (Ib):

<img file="PL2880035T3_D0003.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro; n is a number from zero to 4;

R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and -NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C1-6 alkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl; a means one or two;

Ring A is an aromatic ring;

R²⁰ and R²¹ are independently selected from hydrogen and C 1-6 alkyl; wherein the alkyl is unsubstituted or substituted with amino, hydroxyl, or halo; where R²¹ it may be absent depending on the needs to satisfy the valence;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, C1-6 acyl, SO2-C1-6alkyl, C3-7 cycloalkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or Hello;

or a pharmaceutically acceptable salt thereof.

[0009] The present disclosure provides a compound of Formula (II):

R¹ ABOUT

<img file="PL2880035T3_D0004.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl; R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro; n is a number from zero to 4;

R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and -NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C1-6 alkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0010] The present disclosure provides a compound of Formula (III):

<img file="PL2880035T3_D0005.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro; n is a number from zero to 4;

R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and -NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C1-6 alkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0011] The present disclosure provides a compound of Formula (IV):

<img file="PL2880035T3_D0006.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro; n is a number from zero to 4;

R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and -NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C1-6 alkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0012] The present disclosure provides a compound of Formula (V):

<img file="PL2880035T3_D0007.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0013] The present disclosure provides a compound of Formula (VI):

<img file="PL2880035T3_D0008.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0014] In certain embodiments, the present disclosure provides a compound of Formula (VII) as described below.

[0015] In some embodiments, the compound of Formula (I) - (VIII) is a compound selected from these kinds, described or illustrated in detail in the description below.

[0016] In a further aspect, the present disclosure provides a pharmaceutical composition comprising at least one compound of Formula (I) - (VIII) or a pharmaceutically acceptable salt thereof. The pharmaceutical compositions may additionally contain a pharmaceutically acceptable carrier or excipient. The present disclosure also provides a compound of Formula (I) - (VIII) or a pharmaceutically acceptable salt thereof for use as a medicament.

[0017] In another aspect, the present disclosure provides a compound of Formula (I) - (VIII) or a pharmaceutically acceptable salt thereof for use in therapy. In another aspect, the present disclosure provides a compound of Formula (I) - (VIII) or a pharmaceutically acceptable salt thereof for use in the treatment of a proliferative disorder. In another aspect, the present disclosure provides the use of a compound of Formula (I) - (VIII) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a proliferative disorder.

[0018] The compounds described herein can treat a condition that is associated with EGFR inhibitory activity directed to EGFR mutant but not wild-type EGFR. The compounds of the invention may be administered to a subject in need of such treatment for an effective amount of at least one compound of Formula (I) - (VIII) or a pharmaceutically acceptable salt thereof. In this regard, the mutant EGFR may contain the T790M mutation. The present disclosure provides the use of a compound of Formula (I) - (VIII) in the manufacture of a medicament for the treatment of such diseases and medical conditions and the use of these compounds and salts in the treatment of such diseases and medical conditions.

[0019] The compounds of the present invention may be inhibited in a cell by mutant EGFR, wherein the cell is contacted with an effective amount of at least one compound of Formula (I) - (VIII) or salt thereof, and / or with at least one pharmaceutical composition of the Examples wherein the contacting is in vitro, ex vivo, or in vivo. Therefore, the mutant EGFR may contain the T790M mutation.

[0020] The compounds of the present invention may treat a disease or medical condition associated with kinase inhibitory activity, wherein an individual in need of such treatment is administered an effective amount of at least one compound of Formula (I) - (VIII) or a pharmaceutically acceptable salt thereof, wherein the kinase can be selected from the group consisting of EGFR, EGFR (T790M), BLK, BMX / ETK, BTK, JAK2, JAK3, TEC, TXK, FLT3, and FLT3 (D835Y). The compound of Formula (I) - (VIII) can also be used in the manufacture of a medicament for the treatment of such diseases and medical conditions, and the compounds and salts can be used to treat such diseases and medical conditions. The compounds of the invention may be inhibited by a mutant kinase in cells, wherein the cell is contacted with an effective amount of at least one compound of Formula (I) - (VIII) or a salt thereof, and / or at least one pharmaceutical composition according to embodiments, wherein the contacting takes place in vitro, ex vivo, or in vivo. Accordingly, the mutant kinase can be FLT3 with the D835Y mutation.

One skilled in the art will recognize that the compounds of Formula (II) - (VI) are compounds of Formula (I), and that compounds of Formula (I) - (VII) are compounds of Formula (VIII).

[0022] Additional embodiments, features and advantages of the invention will become apparent from the following detailed description and practical implementation of the invention.

Brief Description of the Drawings [0023]

Figure 1 shows the SDS-PAGE of some effectors of lung cancer H1975 cells treated with various concentrations of Compound 3.

Figure 2 shows immunoblots of certain effectors in Compound 3 treated tumors at various time intervals.

Figure 3 shows a graph of changes in body weight of mice in different groups in the NCI-H1975 model.

Figure 4 shows a graph of changes in body weight of mice in different groups in the HCC827 model.

Figure 5 is a graph of changes in body weight of mice in different groups in the A431 model.

Figure 6 shows a plot of tumor volume in mice in different groups in the NCIH1975 model.

Figure 7 is a plot of tumor volume in mice in different groups in the HCC827 model.

Figure 8 is a plot of tumor volume in mice in different groups in the A431 model.

Figures 9A-9F show SDS-PAGE (9A, 9C, 9E) and inhibition graphs (9B, 9D, 9F) of EGFR-Tyr1068 phosphorylation and downstream signaling in H1975 lung tumor cells treated with various concentrations of Compound 3 (9A, 9B), Gefitinib ( 9C, 9D), and WZ4002 (9E, 9F).

Figures 10A-10F show SDS-PAGE (10A, 10C, 10E) and inhibition graphs (10B, 10D, 10F) of EGFR-Tyr1068 phosphorylation and downstream signaling in cells with mutant EGFR HCC-827 treated with various concentrations of Compound 3 (10A, 10B) , Gefitinib (10C, 10D), and WZ4002 (10E, 10F).

Figures 11A-11F show SDS-PAGE (11A, 11C, 11E) and inhibition graphs (11B, 11D, 11F) of EGFR-Tyr1068 phosphorylation and downstream signaling in A431 cells expressing WT EGFR, which were treated with various concentrations of Compound 3 (11A, 11B) , Gefitinib (11C, 11D), and WZ4002 (11E, 11F).

Figure 12 shows the inhibition of EGFR phosphorylation in tumor tissues of H1975 when treated with a single dose of Compound 3 at 12.5, 50, and 200 mg / kg.

Figure 13 shows the inhibition of EGFR phosphorylation in tumor tissues of H1975 when treated with eight doses of Compound 3 at 12.5 and 50 mg / kg, compared to Gefitinib at 100 mg / kg.

Figure 14 shows the results of a cell-based "pulse-chase" assay that show Compound 3 is an irreversible inhibitor of H1975 cell proliferation with the T790M EGFR mutation.

Figure 15A shows the IC 50 titration curve for Compound 3 against BLK. Figure 15B shows the IC50 titration curve for staurosporin against BLK

Figure 16 shows the IC50 titration curve for Compound 3 against BMX / ETK.

Figure 17 shows the IC 50 titration curve for Compound 3 against BTK.

Figure 18 shows the IC50 titration curve for Compound 3 against FLT3 (D835Y).

Figure 19 shows the IC50 titration curve for Compound 3 against ITK. Figure 20 shows the IC50 titration curve for Compound 3 against JAK2. Figure 21 shows the IC50 titration curve for Compound 3 against JAK3. Figure 22 shows the IC 50 titration curve for Compound 3 against TEC. Figure 23 shows the IC 50 titration curve for Compound 3 against TXK.

Detailed Description [0024] The present invention is directed to a certain piropyropyrimine derivative, pharmaceutical compositions and such compounds and compositions for use in the treatment of proliferative disorders. The compounds described herein have anti-tumor, anti-tumor, anti-inflammatory, anti-infective and anti-proliferative activity. In some embodiments, the compounds have been shown to have antitumor activity in cell-based assays as described herein, using a variety of tumor cell lines that exhibit very effective EGFR inhibitory activity directed essentially on the mutation and essentially not on the wild-type protein. In some cases, the mutant EGFR contains the T790M mutation. According to this, the compounds and compositions containing these compounds of the embodiments are useful in treating conditions characterized by these mutant tumor cells. In some cases, the compounds are useful for the treatment of sarcoma, epidermoid tumor, fibrosarcoma, cervical tumor, gastric cancer, skin cancer, leukemia, lymphoma, lung cancer, non-small cell lung cancer, colon cancer, CNS tumor, melanoma, ovarian cancer, cancer kidney, prostate cancer, breast cancer, liver cancer, head and neck cancer and pancreatic cancer.

[0025] In other embodiments, these compounds have been found to have activity on a number of protein kinases, including EGFR, EGFR (T790M), BLK, BMX / ETK, BTK, JAK2, JAK3, TEC, TXK, FLT3, and FLT3 (D835Y ). In some cases, these compounds are useful for the treatment of tumors, tumors, inflammatory diseases, autoimmune diseases or immunosuppressed diseases. In other embodiments, such diseases are mediated by at least one kinase selected from BTK, JAK3, ITK, and BMX. In other embodiments, tumors, tumors, inflammatory diseases, autoimmune diseases or immunosuppressed diseases mediate malfunctioning B lymphocytes, T lymphocytes, or both. In other embodiments, inflammatory diseases, autoimmune diseases, or immunosuppressed diseases are arthritis, rheumatoid arthritis, lung cancer, non-small cell lung cancer, colon cancer, CNS tumor, melanoma, ovarian cancer, kidney cancer, prostate cancer, breast cancer, liver cancer, head and neck cancer, or pancreatic cancer. In other embodiments, the diseases are autoimmune diseases or inflammatory disorders induced by graft including, but not limited to, allograft, graft versus host disease, or autoimmune diabetes.

[0026] Before the present invention is further described, it should be understood that this invention is not limited to the specific embodiments described, as such may of course vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0027] It should be noted that, as used in the specification and in the appended claims, the singular forms "a", "a" and "the" include plural references unless the context clearly dictates otherwise. It should further be noted that the claims can be made with the exclusion of any eventual element. As such, this statement is intended to serve as a precursor of the basis for the use of such exclusive terminology as "only", "only" and the like, in conjunction with reference to the elements of the claim, or to the use of a "negative" restriction.

[0028] As used herein, the terms "including", "comprising" and "including" are used in their open, non-limiting sense.

[0029] In order to provide a more concise description, some of the quantitative expressions given herein are not qualified with the term "about". It is to be assumed that regardless of whether the term "about" is used or not, each number given herein means a reference to the actually given value and also means a reference to an approximation to such a value that would reasonably be deduced from ordinary skill in this field of technology, including equivalents and approximations due to experimental and / or measurement conditions for the given value. Whenever the yield is given in percent, this yield refers to the mass of the substance for which this yield is given with respect to the maximum amount of the same substance that could be obtained under certain stoichiometric conditions.

[0030] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although all methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, preferred methods and materials are described. All publications mentioned herein are hereby incorporated by reference to disclose and describe the methods and / or materials for which publications are cited.

[0031] Unless otherwise indicated, the methods and techniques of the present embodiments are typically performed in accordance with conventional methods well known in the art or described in various general and more specific references that are cited or discussed herein. See, for example, Loudon, Organic Chemistry, Fourth Edition, New York: Oxford University Press, 2002; Smith and 0March, March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Fifth Edition, Wiley-Interscience, 2001.

[0032] The nomenclature used herein to name the subject compounds is set forth herein in the Examples. This nomenclature was generally derived using commercially available AutoNom software (MDL, San Leandro, Calif.).

[0033] It will be appreciated that certain features of the invention, which for clarity are described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which for the sake of brevity are described in the context of one embodiment, may also be provided separately or in any suitable subcombination. All combinations of embodiments related to the chemical groups represented by the variables are specifically encompassed by the present invention and are disclosed herein as if each combination were individually and specifically disclosed to the extent that such combinations include compounds that are stable compounds (i.e. that can be isolated, characterized and tested for biological activity). Additionally,

Chemical Definitions [0034] The term "alkyl" refers to a straight or branched chain alkyl group containing from 1 to 12 carbon atoms in the chain. Examples of alkyl groups include methyl (Me), ethyl (Et), n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl (tBu), pentyl, isopentyl, tert-pentyl, hexyl, isohexyl, and groups, which in the light of the general knowledge in the art and the information contained herein would be considered equivalent to any of the above examples.

[0035] The term "alkoxy" refers to an alkyl group as defined above linked to an oxygen atom. The alkoxy group is connected to the parent structure via an oxygen atom.

[0036] The term "amino" refers to the group -NH2 or a mono- or dialkylamino group. [0037] The term "cycloalkyl" refers to a saturated or partially saturated, monocyclic, fused polycyclic, bridged polycyclic or spiro polycyclic carbocycle having from 3 to 12 ring atoms on a carbocycle. Illustrative examples of cycloalkyl groups include the following units in the form of correctly bound moieties:

<a name="caption2"></a>&gt;. noO OO o. 0.0.0.

CO. 03.00.03.

n&gt; .0&gt;. JO O · dY / 0.

The term "heteroaryl" refers to a monocyclic, fused bicyclic or condensed polycyclic aromatic heterocycle (a ring structure having ring atoms selected from carbon atoms and up to four heteroatoms selected from nitrogen, oxygen and sulfur) having from 3 to 12 ring atoms. on heterocycle. Illustrative examples of heteroaryl groups include the following units in the form of correctly bound moieties:

<img file="PL2880035T3_D0009.tif" />

<img file="PL2880035T3_D0010.tif" />

<img file="PL2880035T3_D0011.tif" />

<img file="PL2880035T3_D0012.tif" />

<img file="PL2880035T3_D0013.tif" />

[0039] The term "halogen" means chloro, fluoro, bromo or iodo. The term "halo" means chloro, fluoro, bromo, or iodo. The term "haloalkyl" means an alkyl as defined above substituted with one or more halogen atoms. The term "haloalkoxy" means alkoxy as defined above, substituted with one or more halogen atoms.

[0040] The term "acyl" refers to the group RC (O) - from 1 to 10 carbon atoms with a straight or branched or cyclic configuration or a combination thereof, attached to the parent structure via a carbonyl functional group. Such groups may be saturated or unsaturated, and aliphatic or aromatic.

[0041] The term "cyano" refers to the group -CN.

[0042] The term "nitro" refers to the group -NO2.

[0043] The term "hydroxyl" refers to the group -OH.

[0044] Those skilled in the art will recognize that the forms mentioned or indicated above are not exhaustive and that other forms within the scope of these defined terms may also be selected.

[0045] The term "substituted" means that a particular group or moiety has one or more substituents. The term "unsubstituted" means that the specified group has no substituents. The term "optionally substituted" means that the specified group is unsubstituted or substituted by one or more substituents. When the term "substituted" is used to describe a structural system, it is understood that the substitution occurs at any position with acceptable valence on the system.

[0046] Any formula depicted herein is intended to mean a compound with this structural formula, as well as certain embodiments or forms. For example, the formula provided herein is intended to include a racemic form, or one or more enantiomeric, diastereomeric or geometric isomers, or mixtures thereof. In addition, any formula given herein is also intended to refer to a hydrate, solvate or polymorph of such a compound, or a mixture thereof.

[0047] Any of the formulas given herein is also intended to designate unlabeled forms as well as forms of isotopically-labeled compounds. Isotopically labeled compounds have the structures depicted in the formulas given herein, except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into compounds of the embodiments, include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine and iodine, such as²H ³H ¹¹C ¹³C ¹⁴C ¹⁵N ¹⁸ABOUT, ¹⁷ABOUT, ³¹P ³²P ³⁵S ¹⁸F ³⁶Cl, i ¹²⁵I. Such isotopically labeled compounds are useful in metabolic studies (preferably from ¹⁴C), kinetic studies of reactions, (e.g. ²H or ³H), detection or imaging techniques [such as positron emission tomography (PET) or single photon emission computed tomography (SPECT)], including tissue or substrate tissue distribution tests or in radioactive treatment of patients. In particular, a labeled compound¹⁸F or ¹¹C may be particularly beneficial in PET or SPECT studies. Further, substitution with heavier isotopes such as deuterium (i.e.²H) may afford certain therapeutic advantages resulting from greater metabolic stability, e.g. an increased in vivo half-life or reduced dosage requirements. Isotopically labeled compounds of the embodiments and their prodrugs can be generally prepared by carrying out the procedures disclosed in the Schemes or in the Examples and Methods described below, by replacing readily available isotopically labeled reagent with isotopically labeled reagent.

[0048] The nomenclature "Ci-j" zj&gt; i, when used herein for a class of substituents, is intended to refer to embodiments in which each one of the plurality of carbon members from i to j, including iij, is performed independently of one another. . By way of example, the term C 1-3 refers independently to embodiments having one carbon (C 1) member, embodiments having two carbon (C 2) members, and embodiments having three carbon (C 3) members.

[0049] Any disubstitute referred to herein is intended to include various attachment options when more than one of these possibilities is allowed. For example, a reference to the dis-substituent -AB-, wherein A ψ B, refers to a disubstituted A attached to the first substituted member and B attached to a second substituted member and also refers to such a disubstituted A attached to the second substituted member and B attached to the first substituted member.

[0050] The present disclosure provides pharmaceutically acceptable salts of the compounds of Formulas (I) - (VIII), preferably those described above, and the specific compounds described herein, and pharmaceutical compositions comprising such salts and methods of using such salts.

[0051] A "pharmaceutically acceptable salt" is defined as the salt of the free acid or base of a compound shown herein that is non-toxic, biologically acceptable or otherwise biologically suitable for administration to a subject. See, in general, SM Berge, et al., "Pharmaceutical Salts", J. Pharm. Sci., 1977, 66, 1-19. Preferred pharmaceutically acceptable salts are those that are pharmacologically effective and suitable for contact with the tissues of patients without excessive toxicity, irritation or allergic response. The compound described herein can have a sufficiently acidic group, a sufficiently basic group, both types of functional groups, or more than one of each type, and therefore react with a variety of inorganic or organic bases and inorganic and organic acids,

Examples of pharmaceutically acceptable salts include sulphates, metabisulfites, hydrogen sulphates, sulfites, bisulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanes, propiolates, oxalates, malonates, succinates, suberiods, sebacates, fumarates, maleates, butine-1,4-dianates, hexyno-1,6-dionates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulphonates, methylsulfonates, propylsulfonates, benzenesulfonates, xylene sulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, phenylacetates, phenylpropionates, phenylbutyrate, citrates, lactates, gamma-hydroxybutyrate, glycolates, tartrates, and mandelanes.

like mandelic acid, citric acid or tartaric acid, an amino acid such as aspartic acid or glutamic acid, an aromatic acid such as benzoic acid, 2-acetoxybenzoic acid, naphthoic acid or cinnamic acid, a sulphonic acid such as lauryl sulphonic acid, p-acid toluenesulfonic acid, methanesulfonic acid or ethanesulfonic acid, or any compatible mixture of acids such as those exemplified herein and any other acid and mixture thereof, which are considered equivalents or acceptable substitutes in view of the usual skill level in the field of this technology . In some embodiments, the pharmaceutically acceptable salt is a hydrochloride, a maleic acid salt, an HBr salt, a hydroxybutanedioic acid salt, a fumaric acid salt, a lactic acid salt,

[0054] The present disclosure provides pharmaceutically acceptable prodrugs of the compounds of Formulas (I) - (VIII), and their use in therapeutic treatment. The term "prodrug" means a precursor of a designated compound that, when administered to a subject, yields the compound in vivo in a chemical or physiological process such as solvolysis or enzymatic cleavage, or under physiological conditions (e.g., the prodrug, when brought to physiological pH, is converted to a compound of Formulas (I) - (VIII)). A "pharmaceutically acceptable prodrug" is a prodrug that is non-toxic, biologically acceptable or otherwise biologically suitable for administration to a subject. Pictorial procedures for the selection and preparation of appropriate prodrug derivatives are described, for example, in "Design of Prodrugs", ed. H. Bundgaard, Elsevier, 1985.

[0055] The present disclosure provides pharmaceutically active metabolites of compounds with

Formulas (I) - (VIII), and the use of such metabolites in the methods of the embodiments. "Pharmaceutically active metabolite" means the pharmacologically active product of a metabolism in the body of a compound of Formulas (I) - (VIII) or a salt thereof. Prodrugs and active metabolites of the compound can be determined using conventional techniques known and available in the art. See, e.g., Bertolini et al., J. Med. Chem. 1997, 40, 2011-2016; Shan et al., J. Pharm. Sci. 1997, 86 (7), 765-767; Bagshawe, Drug Dev. Res. 1995, 34, 220-230; Bodor, Adv. Drug Res. 1984, 13, 255-331; Bundgaard, Design of Prodrugs (Elsevier Press, 1985); and Larsen, Design and Application of Prodrugs, Drug Design and Development (Krogsgaard-Larsen et al., eds., Harwood Academic Publishers, 1991).

Representative Practice Examples

Formula (VIII) [0056] The present disclosure provides a compound of Formula (VIII). In some embodiments, X¹ means O or NH. In other embodiments, X¹ means CH2 or CF2. In still other embodiments, X¹ means O.

[0057] In some embodiments of Formula (VIII), -NR¹⁸R¹⁹ means

<img file="PL2880035T3_D0014.tif" />

In other embodiments, -NR¹⁸R¹⁹ means yC r¹⁵-ⁿL^nh

In some embodiments, R¹⁵ means methyl, hydroxyethyl, methoxyethyl, or fluoroethyl. In other embodiments, R¹⁵ means fluoroethyl. In some embodiments, m is 1. In other embodiments, m is 2.

[0058] In some embodiments, R⁹ and R¹⁹ taken together form an optionally substituted 5- or 6-membered heteroaryl ring. In some embodiments, R¹⁹ and R⁹ together form a 5- or 6-membered ring optionally substituted with C 16 alkyl which is unsubstituted or substituted with amino. In some embodiments, the heteroaryl ring is substituted with dimethylaminomethyl or piperidinylmethyl. In other embodiments, R⁹ and R¹⁹ taken together, they form pyrrole or pyridine. In some embodiments, R¹⁸ means dimethylaminoethyl.

[0059] In some embodiments, R⁶ means methoxy. In other embodiments, R⁷ means methoxy. In some cases, R⁷ means hydrogen or methoxy.

[0060] In certain embodiments of Formula (VIII), each variable therein is determined as described below for any of Formulas (I) - (VII) or their embodiments. In particular, certain embodiments of Formula (VIII) are as defined for each variable for Formula (I) below, and said definitions are incorporated herein by reference.

Formula (I) [0061] The present disclosure provides a compound of Formula (Ia) and (Ib):

<img file="PL2880035T3_D0015.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro; n is a number from zero to 4;

R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and -NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C1-6 alkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

Q is CR⁹ or N;

where R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl;

a means one or two;

Ring A is an aromatic ring;

R²⁰ and R²¹ are independently selected from hydrogen and C 1-6 alkyl; wherein the alkyl is unsubstituted or substituted with amino, hydroxyl, or halo; where R²¹ may not be present to satisfy the valence;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, C1-6 acyl, SO2-C1-6alkyl, C3-7 cycloalkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or Hello;

or a pharmaceutically acceptable salt thereof.

[0062] Formula (I) is meant to refer to Formula (Ia) and Formula (Ib).

[0063] In Formula (I), R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl. In some cases, R¹ means hydrogen. In some cases, R¹is C1-6 alkyl. In some cases, R¹ means methyl or ethyl. In some cases, R² means hydrogen. In some cases, R² is C1-6 alkyl. In some cases, R² means methyl or ethyl. In some cases, R¹ and R² both are hydrogen.

[0064] In Formula (I), n is a number from zero to 4. In some cases, n is zero. In some cases, n is one. In some cases, n means 2. In some cases, n means 3. In some cases, n means 4.

[0065] In Formula (I), R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro. In some cases, R³ means halo. In some cases, R³means hydroxyl. In some cases, R³ is C1-6 alkyl. In some cases, R³ means C1-6 alkoxy. In some cases, R³ means cyano. In some cases, R³ means nitro.

[0066] In Formula (I), R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and NR²²R²³; wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²²and R²¹ may be combined to form a 3 to 10 membered ring. In some cases, R⁴ means hydrogen. In some cases, R⁴ is C1-6 alkyl. In some cases, R⁴ means C3-7 cycloalkyl. In some cases, R⁴ means NR²²R²³.

[0067] In some cases, R⁴ is unsubstituted C 1-6 alkyl. In some cases, R⁴ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R⁴ is C1-3 alkyl which is substituted with hydroxyl. In some cases, R⁴ is C1-6 alkyl which is substituted with an amino. In some cases,_R4 is C1-6 alkyl which is substituted with -NH2. In some cases, R⁴ is C16 alkyl which is substituted with -N (CH3) 2. In some cases, R⁴ is C1-3 alkyl which is substituted with -NH2. In some cases, R⁴ is C1-3 alkyl which is substituted with -N (CH3) 2.

[0068] In some cases, R⁴ is unsubstituted C3-7 cycloalkyl. In some cases, R⁴ means unsubstituted C3 cycloalkyl. In some cases, R⁴is unsubstituted C4 cycloalkyl. In some cases, R⁴ is unsubstituted C5-6 cycloalkyl. In some cases, R⁴ means unsubstituted C7 cycloalkyl.

[0069] In some cases, R⁴ means -NR²²R²³, each R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be combined to form a 3 to 10 membered ring. In some cases, R²² and R²³ means hydrogen. In some cases, R²² and R²³ means C1-6 alkyl. In some cases, R²² and R²³ means C1-3 alkyl. In some cases, R²² and R²³ means methyl.

[0070] In some cases, R²² and R²³ may be joined to form a 3 to 10 membered ring such that R &amp; lt; 1 &amp; gt;⁴ means

<img file="PL2880035T3_D0016.tif" />

where w is a number from 1 to 8. In some cases, R²² and R²³ may be combined to form a 3-membered ring. In some cases, R²² and R²³ may be combined to form a 4-membered ring. In some cases, R²² and R²³may be combined to form a 5-membered ring. In some cases, R²² and R²³ may be combined to form a 6-membered ring. In some cases, R²² and R²³ may be combined to form a 7-membered ring.

[0071] In Formula (I), R⁵ is selected from hydrogen and C1-6 alkyl. In some cases, R⁵ means hydrogen. In some cases, R⁵ is C1-6 alkyl. In some cases, R⁵ means methyl. In some cases, R⁵ means ethyl. In some cases, R⁵ is C1-3 alkyl. In some cases, R⁵ is C4-6 alkyl.

[0072] In Formula (I), R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁶ means hydrogen. In some cases, R⁶ means halo. In some cases, R⁶ means fluoro. In some cases, R⁶ means chloro. In some cases, R⁶ means bromo. In some cases, R⁶ is C1-6 alkyl. In some cases, R⁶means C1-6 haloalkyl. In some cases, R⁶ means C2-6 alkoxy. In some cases, R⁶ means C1-6 haloalkoxy. In some cases, R⁶ means hydroxyl. In some cases, R⁶ means cyano. In some cases, R⁶ means nitro.

[0073] In Formula (I), R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁷ means hydrogen. In some cases, R⁷ means halo. In some cases, R⁷ means fluoro. In some cases, R⁷ means chloro. In some cases, R⁷ means bromo. In some cases, R⁷ is C1-6 alkyl. In some cases, R⁷means C1-6 haloalkyl. In some cases, R⁷ means C2-6 alkoxy. In some cases, R⁷ means C1-6 haloalkoxy. In some cases, R⁷ means hydroxyl. In some cases, R⁷ means cyano. In some cases, R⁷ means nitro.

[0074] In Formula (I), R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁸ means hydrogen. In some cases, R⁸ means halo. In some cases, R⁸ means fluoro. In some cases, R⁸ means chloro. In some cases, R⁸ means bromo. In some cases, R⁸ is C1-6 alkyl. In some cases, R⁸means C1-6 haloalkyl. In some cases, R⁸ means C1-6 alkoxy. In some cases, R⁸ means C1-6 haloalkoxy. In some cases, R⁸ means hydroxyl. In some cases, R⁸ means cyano. In some cases, R⁸ means nitro.

[0075] In Formula (I), Q is CR⁹ or N. In some cases, Q is CR⁹. In some cases, Q is N.

[0076] In Formula (I), R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁹ means hydrogen. In some cases, R⁹ means halo. In some cases, R⁹ means fluoro. In some cases, R⁹ means chloro. In some cases, R⁹ means bromo. In some cases, R⁹ is C1-6 alkyl. In some cases, R⁹means C1-6 haloalkyl. In some cases, R⁹ means C1-6 alkoxy. In some cases, R⁹ means C1-6 haloalkoxy. In some cases, R⁹ means hydroxyl. In some cases, R⁹ means cyano. In some cases, R⁹ means nitro. In some cases, R⁹ means hydrogen or fluoro.

[0077] In Formula (I), R¹⁰ is selected from hydrogen and C1-6 alkyl. In some cases, R¹⁰ means hydrogen. In some cases, R¹⁰ is C1-6 alkyl. In some cases, R¹⁰ means methyl. In some cases, R¹⁰ means ethyl. In some cases, R¹⁰ is C1-3 alkyl. In some cases, R¹⁰ is C4-6 alkyl.

[0078] In Formula (I), a is one or two and Ring A is an aromatic ring. In some cases, a means one, as shown:

<img file="PL2880035T3_D0017.tif" />

In some cases, a means two, as shown:

<img file="PL2880035T3_D0018.tif" />

[0079] In Formula (I), R²⁰ and R²¹ are independently selected from hydrogen and C 1-6 alkyl; wherein the alkyl is unsubstituted or substituted with amino, hydroxyl, or halo; where R²¹ may not be present to satisfy the valence. In some cases, R²⁰ and R²¹ they are independently hydrogen. In some cases, R²⁰ and R²¹ independently are unsubstituted C 1-6 alkyl. In some cases, R²⁰ and R²¹ independently are C 1-6 alkyl, substituted amino. In some cases, R²⁰ and R²¹ independently are C 1-6 alkyl, substituted with NR²⁴R²⁵with R²⁴ and R²⁵ are independently selected from hydrogen and C 1-6 alkyl. In some cases, R²⁰ and R²¹ independently are C 1-6 alkyl, substituted with -NR²⁴R²⁵with R²⁴and R²⁵ are independently selected from hydrogen and C 1-3 alkyl. In some cases, R²⁰ and R²¹independently are C 1-6 alkyl, substituted with -NR²⁴R²⁵with R²⁴ and R²⁵ are independently selected from hydrogen and methyl. In some cases, R²⁰ and R²¹ independently are C 1-3 alkyl, substituted with -NR²⁴R²⁵with R²⁴ and R²⁵ are independently selected from hydrogen and methyl. In some cases, R²⁰ and R²¹ independently are C 1-6 alkyl substituted with hydroxy. In some cases, R²⁰ and R²¹ means C1-6 alkyl, substituted halo. [0080] In Formula (I), R¹¹ is selected from hydrogen and C1-6 alkyl. In some cases, R¹¹ means hydrogen. In some cases, R¹¹ is C1-6 alkyl. In some cases, R¹¹ means methyl. In some cases, R¹¹ means ethyl. In some cases, R¹¹ is C1-3 alkyl. In some cases, R¹¹ is C4-6 alkyl.

[0081] In Formula (I), R¹² is selected from hydrogen and C1-6 alkyl. In some cases, R¹² means hydrogen. In some cases, R¹² is C1-6 alkyl. In some cases, R¹² means methyl. In some cases, R¹² means ethyl. In some cases, R¹² is C1-3 alkyl. In some cases, R¹² is C4-6 alkyl.

[0082] In Formula (I), R¹³ is selected from hydrogen, C1-6 alkyl, C1-6 acyl, SO2-C1-6alkyl, C3-7 cycloalkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or Hello. In some cases, R¹³ means hydrogen. In some cases, R¹³ is C1-6 alkyl. In some cases, R¹³means C1-6 acyl. In some cases, R¹³ means SO2-C1-6alkyl. In some cases, R¹³ means C3-7 cycloalkyl. In some cases, R¹³ means C6-20 aryl. In some cases, R¹³ is C1-6alkyl substituted with hydroxyl or halo.

[0083] In some cases, R¹³ is unsubstituted C 1-6 alkyl. In some cases, R¹³ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R¹³ means - (CH2) mOH, where m is a number from one to 3. In some cases, R¹³ means -CH2OH. In some cases, R¹³ is C1-6 alkyl which is substituted with C1-6 alkoxy.

[0084] In some cases, R¹³ is C1-6 alkyl which is substituted with halo. In some cases, R¹³ is - (CH2) mX, wherein m is a number from one to three and X is halo. In some cases, R¹³ is C1-6 alkyl which is substituted with fluoro. In some cases, R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R¹³ means - (CH2) 2F.

[0085] In some cases, R¹³ means C1-6 acyl. In some cases, R¹³means C1 acyl. In some cases, R¹³ means C2 acyl. In some cases, R¹³ means C3 acyl. In some cases, R¹³ means C4-6 acyl.

[0086] In some cases, R¹³ means SO2-C1-6alkyl. In some cases, R¹³means SO2-C1alkyl. n some examples, R¹³ means SO2-C2alkyl. In some cases, R¹³ means SO2-C3alkyl. In some cases, R¹³ means SO2-C4-6alkyl. [0087] In some cases, R¹³ means C3-7 cycloalkyl. In some cases, R¹³means unsubstituted C3 cycloalkyl. In some cases, R¹³ is unsubstituted C4 cycloalkyl. In some cases, R¹³ is unsubstituted C5-6 cycloalkyl. In some cases, R¹³ means unsubstituted C7 cycloalkyl.

[0088] In some cases, R¹³ means unsubstituted C6-20 aryl. In some cases, R¹³ means C6-20 aryl which is substituted with hydroxyl. In some cases, R¹³ means C6-20 aryl which is substituted with C1-6 alkoxy. In some cases, R¹³ means C6-20 aryl which is substituted with halo.

[0089] In some cases, R⁶, R⁷, R⁸and R⁹ means hydrogen. In some cases, R⁶, R⁷and R⁸ are hydrogen and R⁹ means halo. In some cases, R⁶, R⁷and R⁸ are hydrogen and R⁹ means fluoro.

[0090] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹⁰ means methyl. In some cases, R⁶, R⁷, R⁸ means hydrogen; R⁹ means halo; and R¹⁰ means methyl. In some cases, R⁶, R⁷, R⁸ means hydrogen; R⁹ means fluoro; and R¹⁰ means methyl. [0091] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means hydrogen.

[0092] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means -CH2OH. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means -CH2OH. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means CH2OH.

[0093] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means (CH2) mF, where m is a number from one to three.

Formula (II) [0094] The present disclosure provides a compound of Formula (II):

<img file="PL2880035T3_D0019.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro; n is a number from zero to 4;

R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and -NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²¹may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C1-6 alkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

Q is CR⁹ or N;

where R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0095] In Formula (II), R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 16 haloalkyl. In some cases, R¹ means hydrogen. In some cases, R¹is C1-6 alkyl. In some cases, R¹ means methyl or ethyl. In some cases, R² means hydrogen. In some cases, R² is C1-6 alkyl. In some cases, R² means methyl or ethyl. In some cases, R¹ and R²means hydrogen.

[0096] In Formula (II), n is a number from zero to 4. In some cases, n is zero. In some cases, n is one. In some cases, n means 2. In some cases, n means 3. In some cases, n means 4.

[0097] In Formula (II), R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro. In some cases, R³ means halo. In some cases, R³means hydroxyl. In some cases, R³ is C1-6 alkyl. In some cases, R³ means C1-6 alkoxy. In some cases, R³ means cyano. In some cases, R³ means nitro.

[0098] In Formula (II), R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and NR²²R²³; wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²²and R²¹ may be combined to form a 3 to 10 membered ring. In some cases, R⁴ means hydrogen. In some cases, R⁴ is C1-6 alkyl. In some cases, R⁴ means C3-7 cycloalkyl. In some cases, R⁴ means NR²²R²³.

[0099] In some cases, R⁴ is unsubstituted C 1-6 alkyl. In some cases, R⁴ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R⁴ is C1-3 alkyl which is substituted with hydroxyl. In some cases, R⁴ is C1-6 alkyl which is substituted with an amino. In some cases,_R4 is C1-6 alkyl which is substituted with -NH2. In some cases, R⁴ is C16 alkyl which is substituted with -N (CH3) 2. In some cases, R⁴ is C1-3 alkyl which is substituted with -NH2. In some cases, R⁴ is C1-3 alkyl which is substituted with -N (CH3) 2.

[0100] In some cases, R⁴ is unsubstituted C3-7 cycloalkyl. In some cases, R⁴ means unsubstituted C3 cycloalkyl. In some cases, R⁴is unsubstituted C4 cycloalkyl. In some cases, R⁴ is unsubstituted C5-6 cycloalkyl. In some cases, R⁴ means unsubstituted C7 cycloalkyl.

[0101] In some cases, R⁴ means -NR²²R²³, each R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be combined to form a 3 to 10 membered ring. In some cases, R²² and R²³ means hydrogen. In some cases, R²² and R²³ means C1-6 alkyl. In some cases, R²² and R²³ means C1-3 alkyl. In some cases, R²² and R²³ means methyl.

[0102] In some cases, R²² and R²³ may be joined to form a 3 to 10 membered ring such that R &amp; lt; 1 &amp; gt;⁴ means

<img file="PL2880035T3_D0020.tif" />

where w is a number from 1 to 8. In some cases, R²² and R²³ may be combined to form a 3-membered ring. In some cases, R²² and R²³ may be combined to form a 4-membered ring. In some cases, R²² and R²³may be combined to form a 5-membered ring. In some cases, R²² and R²³ may be combined to form a 6-membered ring. In some cases, R²² and R²³ may be combined to form a 7-membered ring.

[0103] In Formula (II), R⁵ is selected from hydrogen and C1-6 alkyl. In some cases,_R5 is hydrogen. In some cases, R⁵ is C1-6 alkyl. In some cases,_R5 is methyl. In some cases, R⁵ means ethyl. In some cases, R⁵is C1-3 alkyl. In some cases, R⁵ is C4-6 alkyl.

[0104] In Formula (II), R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁶ means hydrogen. In some cases, R⁶ means halo. In some cases, R⁶ means fluoro. In some cases, R⁶ means chloro. In some cases, R⁶ means bromo. In some cases, R⁶ is C1-6 alkyl. In some cases, R⁶ means C1-6 haloalkyl. In some cases, R⁶ means C2-6 alkoxy. In some cases, R⁶means C1-6 haloalkoxy. In some cases, R⁶ means hydroxyl. In some cases, R⁶ means cyano. In some cases, R⁶ means nitro.

[0105] In Formula (II), R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁷ means hydrogen. In some cases, R⁷ means halo. In some cases, R⁷ means fluoro. In some cases, R⁷ means chloro. In some cases, R⁷ means bromo. In some cases, R⁷ is C1-6 alkyl. In some cases, R⁷ means C1-6 haloalkyl. In some cases, R⁷ means C2-6 alkoxy. In some cases, R⁷means C1-6 haloalkoxy. In some cases, R⁷ means hydroxyl. In some cases, R⁷ means cyano. In some cases, R⁷ means nitro.

[0106] In Formula (II), R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁸ means hydrogen. In some cases, R⁸ means halo. In some cases, R⁸ means fluoro. In some cases, R⁸ means chloro. In some cases, R⁸ means bromo. In some cases, R⁸ is C1-6 alkyl. In some cases, R⁸ means C1-6 haloalkyl. In some cases, R⁸ means C1-6 alkoxy. In some cases, R⁸means C1-6 haloalkoxy. In some cases, R⁸ means hydroxyl. In some cases, R⁸ means cyano. In some cases, R⁸ means nitro.

[0107] In Formula (II), Q is CR⁹ or N. In some cases, Q is CR⁹. In some cases, Q is N.

[0108] In Formula (II), R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁹ means hydrogen. In some cases, R⁹ means halo. In some cases, R⁹ means fluoro. In some cases, R⁹ means chloro. In some cases, R⁹ means bromo. In some cases, R⁹ is C1-6 alkyl. In some cases, R⁹means C1-6 haloalkyl. In some cases, R⁹ means C1-6 alkoxy. In some cases, R⁹ means C1-6 haloalkoxy. In some cases, R⁹ means hydroxyl. In some cases, R⁹ means cyano. In some cases, R⁹ means nitro.

[0109] In Formula (II), R¹⁰ is selected from hydrogen and C1-6 alkyl. In some cases, R¹⁰ means hydrogen. In some cases, R¹⁰ is C1-6 alkyl. In some cases, R¹⁰ means methyl. In some cases, R¹⁰ means ethyl. In some cases, R¹⁰ is C1-3 alkyl. In some cases, R¹⁰ is C4-6 alkyl.

[0110] In Formula (II), R¹¹ is selected from hydrogen and C1-6 alkyl. In some cases, R¹¹ means hydrogen. In some cases, R¹¹ is C1-6 alkyl. In some cases, R¹¹ means methyl. In some cases, R¹¹ means ethyl. In some cases, R¹¹ is C1-3 alkyl. In some cases, R¹¹ is C4-6 alkyl.

[0111] In Formula (II), R¹² is selected from hydrogen and C1-6 alkyl. In some cases, R¹² means hydrogen. In some cases, R¹² is C1-6 alkyl. In some cases, R¹² means methyl. In some cases, R¹² means ethyl. In some cases, R¹² is C1-3 alkyl. In some cases, R¹² is C4-6 alkyl.

[0112] In Formula (II), R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo. In some cases, R¹³ means hydrogen. In some cases, R¹³ is C1-6 alkyl. In some cases, R¹³ means C6-20 aryl.

[0113] In some cases, R¹³ is unsubstituted C 1-6 alkyl. In some cases, R¹³ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R¹³ means - (CH2) mOH, where m is a number from one to 3. In some cases, R¹³ is-CH2OH. In some cases, R¹³ is C1-6 alkyl which is substituted with C1-6 alkoxy.

[0114] In some cases, R¹³ is C1-6 alkyl which is substituted with halo. In some cases, R¹³ is - (CH2) mX, wherein m is a number from one to three and X is halo. In some cases, R¹³ is C1-6 alkyl which is substituted with fluoro. In some cases, R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R¹³ means - (CH2) 2F.

[0115] In some cases, R¹³ means unsubstituted C6-20 aryl. In some cases, R¹³ means C6-20 aryl which is substituted with hydroxyl. In some cases, R¹³ means C6-20 aryl which is substituted with C1-6 alkoxy. In some cases, R¹³ means C6-20 aryl which is substituted with halo.

[0116] In some cases, R⁶, R⁷, R⁸and R⁹ means hydrogen. In some cases, R⁶, R⁷and R⁸ are hydrogen and R⁹ means halo. In some cases, R⁶, R⁷and R⁸ are hydrogen and R⁹ means fluoro.

[0117] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹⁰ means methyl. In some cases, R⁶, R⁷, R⁸ means hydrogen; R⁹ means halo; and R¹⁰ means methyl. In some cases, R⁶, R⁷, R⁸ means hydrogen; R⁹ means fluoro; and R¹⁰ means methyl. [0118] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means hydrogen.

[0119] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means -CH2OH. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means -CH2OH. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means CH2OH.

[0120] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means (CH2) mF, where m is a number from one to three.

Formula (III) [0121] The present disclosure provides a compound of Formula (III):

<img file="PL2880035T3_D0021.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro; n is a number from zero to 4;

R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and -NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C1-6 alkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0122] In Formula (III), R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl. In some cases, R¹ means hydrogen. In some cases, R¹is C1-6 alkyl. In some cases, R¹ means methyl or ethyl. In some cases, R² means hydrogen. In some cases, R² is C1-6 alkyl. In some cases, R² means methyl or ethyl. In some cases, R¹ and R²means hydrogen.

[0123] In Formula (III), n is a number from zero to 4. In some cases, n is zero. In some cases, n is one. In some cases, n means 2. In some cases, n means 3. In some cases, n means 4.

[0124] In Formula (III), R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro. In some cases, R³ means halo. In some cases, R³means hydroxyl. In some cases, R³ is C1-6 alkyl. In some cases, R³ means C1-6 alkoxy. In some cases, R³ means cyano. In some cases, R³ means nitro.

[0125] In Formula (III), R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and NR²²R²³; wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²²and R²¹ may be combined to form a 3 to 10 membered ring. In some cases, R⁴ means hydrogen. In some cases, R⁴ is C1-6 alkyl. In some cases, R⁴ means C3-7 cycloalkyl. In some cases, R⁴ means NR²²R²³.

[0126] In some cases, R⁴ is unsubstituted C 1-6 alkyl. In some cases, R⁴ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R⁴ is C1-3 alkyl which is substituted with hydroxyl. In some cases, R⁴ is C1-6 alkyl which is substituted with an amino. In some cases,_R4 is C1-6 alkyl which is substituted with -NH2. In some cases, R⁴ is C1-6 alkyl which is substituted with -N (CH3) 2. In some cases, R⁴ is C1-3 alkyl which is substituted with -NH2. In some cases, R⁴ is C1-3 alkyl which is substituted with -N (CH3) 2.

[0127] In some cases, R⁴ is unsubstituted C3-7 cycloalkyl. In some cases, R⁴ means unsubstituted C3 cycloalkyl. In some cases, R⁴is unsubstituted C4 cycloalkyl. In some cases, R⁴ is unsubstituted C5-6 cycloalkyl. In some cases, R⁴ means unsubstituted C7 cycloalkyl.

[0128] In some cases, R⁴ means -NR²²R²³, each R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be combined to form a 3 to 10 membered ring. In some cases, R²² and R²³ means hydrogen. In some cases, R²² and R²³ means C1-6 alkyl. In some cases, R²² and R²³ means C1-3 alkyl. In some cases, R²² and R²³ means methyl.

[0129] In some cases, R²² and R²³ may be joined to form a 3 to 10 membered ring such that R &amp; lt; 1 &amp; gt;⁴ means

<img file="PL2880035T3_D0022.tif" />

where w is a number from 1 to 8. In some cases, R²² and R²³ may be combined to form a 3-membered ring. In some cases, R²² and R²³ may be combined to form a 4-membered ring. In some cases, R²² and R²³ may be combined to form a 5-membered ring. In some cases, R²² and R²³ may be combined to form a 6-membered ring. In some cases, R²² and R²³ may be combined to form a 7-membered ring.

[0130] In Formula (III), R⁵ is selected from hydrogen and C1-6 alkyl. In some cases, R⁵ means hydrogen. In some cases, R⁵ is C1-6 alkyl. In some cases, R⁵ means methyl. In some cases, R⁵ means ethyl. In some cases, R⁵ is C1-3 alkyl. In some cases, R⁵ is C4-6 alkyl.

[0131] In Formula (III), R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 26 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁶ means hydrogen. In some cases, R⁶ means halo. In some cases, R⁶ means fluoro. In some cases, R⁶ means chloro. In some cases, R⁶ means bromo. In some cases, R⁶ is C1-6 alkyl. In some cases, R⁶ means C1-6 haloalkyl. In some cases, R⁶ means C2-6 alkoxy. In some cases, R⁶ means C1-6 haloalkoxy. In some cases, R⁶ means hydroxyl. In some cases, R⁶ means cyano. In some cases, R⁶ means nitro. [0132] In Formula (III), R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁷ means hydrogen. In some cases, R⁷ means halo. In some cases, R⁷ means fluoro. In some cases, R⁷ means chloro. In some cases, R⁷ means bromo. In some cases, R⁷ is C1-6 alkyl. In some cases, R⁷means C1-6 haloalkyl. In some cases, R⁷ means C2-6 alkoxy. In some cases, R⁷ means C1-6 haloalkoxy. In some cases, R⁷ means hydroxyl. In some cases, R⁷ means cyano. In some cases, R⁷ means nitro. [0133] In Formula (III), R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁸ means hydrogen. In some cases, R⁸ means halo. In some cases, R⁸ means fluoro. In some cases, R⁸ means chloro. In some cases, R⁸ means bromo. In some cases, R⁸ is C1-6 alkyl. In some cases, R⁸means C1-6 haloalkyl. In some cases, R⁸ means C1-6 alkoxy. In some cases, R⁸ means C1-6 haloalkoxy. In some cases, R⁸ means hydroxyl. In some cases, R⁸ means cyano. In some cases, R⁸ means nitro. [0134] In Formula (III), R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁹ means hydrogen. In some cases, R⁹ means halo. In some cases, R⁹ means fluoro. In some cases, R⁹ means chloro. In some cases, R⁹ means bromo. In some cases, R⁹ is C1-6 alkyl. In some cases, R⁹means C1-6 haloalkyl. In some cases, R⁹ means C1-6 alkoxy. In some cases, R⁹ means C1-6 haloalkoxy. In some cases, R⁹ means hydroxyl. In some cases, R⁹ means cyano. In some cases, R⁹ means nitro. [0135] In Formula (III), R¹⁰ is selected from hydrogen and C1-6 alkyl. In some cases, R¹⁰ means hydrogen. In some cases, R¹⁰ is C1-6 alkyl. In some cases, R¹⁰ means methyl. In some cases, R¹⁰ means ethyl. In some cases, R¹⁰ is C1-3 alkyl. In some cases, R¹⁰ is C4-6 alkyl.

[0136] In Formula (III), R¹¹ is selected from hydrogen and C1-6 alkyl. In some cases, R¹¹ means hydrogen. In some cases, R¹¹ is C1-6 alkyl. In some cases, R¹¹ means methyl. In some cases, R¹¹ means ethyl. In some cases, R¹¹ is C1-3 alkyl. In some cases, R¹¹ is C4-6 alkyl.

[0137] In Formula (III), R¹² is selected from hydrogen and C1-6 alkyl. In some cases, R¹² means hydrogen. In some cases, R¹² is C1-6 alkyl. In some cases, R¹² means methyl. In some cases, R¹² means ethyl. In some cases, R¹² is C1-3 alkyl. In some cases, R¹² is C4-6 alkyl.

[0138] In Formula (III), R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo. In some cases, R¹³ means hydrogen. In some cases, R¹³ is C1-6 alkyl. In some cases, R¹³ means C6-20 aryl.

[0139] In some cases, R¹³ is unsubstituted C 1-6 alkyl. In some cases, R¹³ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R¹³ means - (CH2) mOH, where m is a number from one to 3. In some cases, R¹³ is-CH2OH. In some cases, R¹³ is C1-6 alkyl which is substituted with C1-6 alkoxy.

[0140] In some cases, R¹³ is C1-6 alkyl which is substituted with halo. In some cases, R¹³ is - (CH2) mX, wherein m is a number from one to three and X is halo. In some cases, R¹³ is C1-6 alkyl which is substituted with fluoro. In some cases, R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R¹³ means - (CH2) 2F.

[0141] In some cases, R¹³ means unsubstituted C6-20 aryl. In some cases, R¹³ means C6-20 aryl which is substituted with hydroxyl. In some cases, R¹³ means C6-20 aryl which is substituted with C1-6 alkoxy. In some cases, R¹³ means C6-20 aryl which is substituted with halo.

[0142] In some cases, R⁶, R⁷, R⁸and R⁹ means hydrogen. In some cases, R⁶, R⁷and R⁸ are hydrogen and R⁹ means halo. In some cases, R⁶, R⁷and R⁸ are hydrogen and R⁹ means fluoro.

[0143] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹⁰ means methyl. In some cases, R⁶, R⁷, R⁸ means hydrogen; R⁹ means halo; and R¹⁰ means methyl. In some cases, R⁶, R⁷, R⁸ means hydrogen; R⁹ means fluoro; and R¹⁰ means methyl.

[0144] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means hydrogen.

[0145] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means -CH2OH. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means -CH2OH. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means CH2OH.

[0146] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means (CH2) mF, where m is a number from one to three.

Formula (IV) [0147] The present disclosure provides a compound of Formula (IV):

<img file="PL2880035T3_D0023.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro; n is a number from zero to 4;

R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and -NR²²R²³;

wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be joined to form a 3 to 10 membered ring;

R⁵ is selected from hydrogen and C1-6 alkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0148] In Formula (IV), R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 16 haloalkyl. In some cases, R¹ means hydrogen. In some cases, R¹ is C1-6 alkyl. In some cases, R¹ means methyl or ethyl. In some cases, R² means hydrogen. In some cases, R² is C1-6 alkyl. In some cases, R² means methyl or ethyl. In some cases, R¹ and R²means hydrogen.

[0149] In Formula (IV), n is a number from zero to 4. In some cases, n is zero. In some cases, n is one. In some cases, n means 2. In some cases, n means 3. In some cases, n means 4.

[0150] In Formula (IV), R³ is selected from halo, hydroxy, C 1-6 alkyl, C 1-6 alkoxy, cyano and nitro. In some cases, R³ means halo. In some cases, R³means hydroxyl. In some cases, R³ is C1-6 alkyl. In some cases, R³ means C1-6 alkoxy. In some cases, R³ means cyano. In some cases, R³ means nitro.

[0151] In Formula (IV), R⁴ is selected from hydrogen, C1-6 alkyl, C3-7 cycloalkyl, and NR²²R²³; wherein the alkyl or cycloalkyl is unsubstituted or substituted by hydroxy or amino; and every R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²²and R²¹ may be combined to form a 3 to 10 membered ring. In some cases, R⁴ means hydrogen. In some cases, R⁴ is C1-6 alkyl. In some cases, R⁴ means C3-7 cycloalkyl. In some cases, R⁴ means NR²²R²³.

[0152] In some cases, R⁴ is unsubstituted C 1-6 alkyl. In some cases, R⁴ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R⁴ is C1-3 alkyl which is substituted with hydroxyl. In some cases, R⁴ is C1-6 alkyl which is substituted with an amino. In some cases,_R4 is C1-6 alkyl which is substituted with -NH2. In some cases, R⁴ is C16 alkyl which is substituted with -N (CH3) 2. In some cases, R⁴ is C1-3 alkyl which is substituted with -NH2. In some cases, R⁴ is C1-3 alkyl which is substituted with -N (CH3) 2.

[0153] In some cases, R⁴ is unsubstituted C3-7 cycloalkyl. In some cases, R⁴ means unsubstituted C3 cycloalkyl. In some cases, R⁴is unsubstituted C4 cycloalkyl. In some cases, R⁴ is unsubstituted C5-6 cycloalkyl. In some cases, R⁴ means unsubstituted C7 cycloalkyl.

[0154] In some cases, R⁴ means -NR²²R²³, each R²² and R²³ are independently selected from hydrogen and C1-6 alkyl or R²² and R²³ may be combined to form a 3 to 10 membered ring. In some cases, R²² and R²³ means hydrogen. In some cases, R²² and R²³ means C1-6 alkyl. In some cases, R²² and R²³ means C1-3 alkyl. In some cases, R²² and R²³ means methyl.

[0155] In some cases, R²² and R²³ may be joined to form a 3 to 10 membered ring such that R &amp; lt; 1 &amp; gt;⁴ means

<img file="PL2880035T3_D0024.tif" />

where w is a number from 1 to 8. In some cases, R²² and R²³ may be combined to form a 3-membered ring. In some cases, R²² and R²³ may be combined to form a 4-membered ring. In some cases, R²² and R²³may be combined to form a 5-membered ring. In some cases,

R²² and R²³ may be combined to form a 6-membered ring. In some cases, R²² and R²³ may be combined to form a 7-membered ring.

[0156] In Formula (IV), R⁵ is selected from hydrogen and C1-6 alkyl. In some cases, R⁵ means hydrogen. In some cases, R⁵ is C1-6 alkyl. In some cases, R⁵ means methyl. In some cases, R⁵ means ethyl. In some cases, R⁵ is C1-3 alkyl. In some cases, R⁵ is C4-6 alkyl.

[0157] In Formula (IV), R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁶ means hydrogen. In some cases, R⁶ means halo. In some cases, R⁶ means fluoro. In some cases, R⁶ means chloro. In some cases, R⁶ means bromo. In some cases, R⁶ is C1-6 alkyl. In some cases, R⁶means C1-6 haloalkyl. In some cases, R⁶ means C2-6 alkoxy. In some cases, R⁶ means C1-6 haloalkoxy. In some cases, R⁶ means hydroxyl. In some cases, R⁶ means cyano. In some cases, R⁶ means nitro.

[0158] In Formula (IV), R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁷ means hydrogen. In some cases, R⁷ means halo. In some cases, R⁷ means fluoro. In some cases, R⁷ means chloro. In some cases, R⁷ means bromo. In some cases, R⁷ is C1-6 alkyl. In some cases, R⁷means C1-6 haloalkyl. In some cases, R⁷ means C2-6 alkoxy. In some cases, R⁷ means C1-6 haloalkoxy. In some cases, R⁷ means hydroxyl. In some cases, R⁷ means cyano. In some cases, R⁷ means nitro.

[0159] In Formula (IV), R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁸ means hydrogen. In some cases, R⁸ means halo. In some cases, R⁸ means fluoro. In some cases, R⁸ means chloro. In some cases, R⁸ means bromo. In some cases, R⁸ is C1-6 alkyl. In some cases, R⁸means C1-6 haloalkyl. In some cases, R⁸ means C1-6 alkoxy. In some cases, R⁸ means C1-6 haloalkoxy. In some cases, R⁸ means hydroxyl. In some cases, R⁸ means cyano. In some cases, R⁸ means nitro.

[0160] In Formula (IV), R¹⁰ is selected from hydrogen and C1-6 alkyl. In some cases, R¹⁰ means hydrogen. In some cases, R¹⁰ is C1-6 alkyl. In some cases, R¹⁰ means methyl. In some cases, R¹⁰ means ethyl. In some cases, R¹⁰ is C1-3 alkyl. In some cases, R¹⁰ is C4-6 alkyl.

[0161] In Formula (IV), R¹¹ is selected from hydrogen and C1-6 alkyl. In some cases, R¹² means hydrogen. In some cases, R¹¹ is C1-6 alkyl. In some cases, R¹¹ means methyl. In some cases, R¹¹ means ethyl. In some cases, R¹¹ is C1-3 alkyl. In some cases, R¹¹ is C4-6 alkyl.

[0162] In Formula (IV), R¹² is selected from hydrogen and C1-6 alkyl. In some cases, R¹² means hydrogen. In some cases, R¹² is C1-6 alkyl. In some cases, R¹² means methyl. In some cases, R¹² means ethyl. In some cases, R¹² is C1-3 alkyl. In some cases, R¹² is C4-6 alkyl.

[0163] In Formula (IV), R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo. In some cases, R¹³ means hydrogen. In some cases, R¹³ is C1-6 alkyl. In some cases, R¹³ means C6-20 aryl.

[0164] In some cases, R¹³ is unsubstituted C 1-6 alkyl. In some cases, R¹³ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R¹³ means - (CH2) mOH, where m is a number from one to 3. In some cases, R¹³ means -CH2OH. In some cases, R¹³ is C1-6 alkyl which is substituted with C1-6 alkoxy.

[0165] In some cases, R¹³ is C1-6 alkyl which is substituted with halo. In some cases, R¹³ is - (CH2) mX, wherein m is a number from one to three and X is halo. In some cases, R¹³ is C1-6 alkyl which is substituted with fluoro. In some cases, R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R¹³ means - (CH2) 2F.

[0166] In some cases, R¹³ means unsubstituted C6-20 aryl. In some cases, R¹³ means C6-20 aryl which is substituted with hydroxyl. In some cases, R¹³ means C6-20 aryl which is substituted with C1-6 alkoxy. In some cases, R¹³ means C6-20 aryl which is substituted with halo.

[0167] In some cases, R⁶, R⁷and R⁸ means hydrogen. In some cases, R⁶, R⁷and R⁸ are hydrogen and R¹⁰ means methyl. In some cases, R⁶, R⁷and R⁸are hydrogen and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ are hydrogen and R¹³ means -CH2OH. In some cases, R⁶, R⁷and R⁸ are hydrogen and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ are hydrogen and R¹³ means - (CH2) mF, wherein m is a number from one to three.

Formula (V) [0168] The present disclosure provides a compound of Formula (V):

<img file="PL2880035T3_D0025.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl;

R¹¹ is selected from hydrogen and C1-6 alkyl;

R¹² is selected from hydrogen and C1-6 alkyl;

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0169] In Formula (V), R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl. In some cases, R¹ means hydrogen. In some cases, R¹is C1-6 alkyl. In some cases, R¹ means methyl or ethyl. In some cases, R² means hydrogen. In some cases, R² is C1-6 alkyl. In some cases, R² means methyl or ethyl. In some cases, R¹ and R²means hydrogen.

[0170] In Formula (V), R⁶ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁶ means hydrogen. In some cases, R⁶ means halo. In some cases, R⁶ means fluoro. In some cases, R⁶ means chloro. In some cases, R⁶ means bromo. In some cases, R⁶ is C1-6 alkyl. In some cases, R⁶means C1-6 haloalkyl. In some cases, R⁶ means C2-6 alkoxy. In some cases, R⁶ means C1-6 haloalkoxy. In some cases, R⁶ means hydroxyl. In some cases, R⁶ means cyano. In some cases, R⁶ means nitro.

[0171] In Formula (V), R⁷ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁷ means hydrogen. In some cases, R⁷ means halo. In some cases, R⁷ means fluoro. In some cases, R⁷ means chloro. In some cases, R⁷ means bromo. In some cases, R⁷ is C1-6 alkyl. In some cases, R⁷means C1-6 haloalkyl. In some cases, R⁷ means C2-6 alkoxy. In some cases, R⁷ means C1-6 haloalkoxy. In some cases, R⁷ means hydroxyl. In some cases, R⁷ means cyano. In some cases, R⁷ means nitro.

[0172] In Formula (V), R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁸ means hydrogen. In some cases, R⁸ means halo. In some cases, R⁸ means fluoro. In some cases, R⁸ means chloro. In some cases, R⁸ means bromo. In some cases, R⁸ is C1-6 alkyl. In some cases, R⁸means C1-6 haloalkyl. In some cases, R⁸ means C1-6 alkoxy. In some cases, R⁸ means C1-6 haloalkoxy. In some cases, R⁸ means hydroxyl. In some cases, R⁸ means cyano. In some cases, R⁸ means nitro.

[0173] In Formula (V), Q is CR⁹ or N. In some cases, Q is CR⁹. In some cases, Q is N.

[0174] In Formula (V), R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁹ means hydrogen. In some cases, R⁹ means halo. In some cases, R⁹ means fluoro. In some cases, R⁹ means chloro. In some cases, R⁹ means bromo. In some cases, R⁹ is C1-6 alkyl. In some cases, R⁹means C1-6 haloalkyl. In some cases, R⁹ means C1-6 alkoxy. In some cases, R⁹ means C1-6 haloalkoxy. In some cases, R⁹ means hydroxyl. In some cases, R⁹ means cyano. In some cases, R⁹ means nitro.

[0175] In Formula (V), R¹⁰ is selected from hydrogen and C1-6 alkyl. In some cases, R¹⁰ means hydrogen. In some cases, R¹⁰ is C1-6 alkyl. In some cases, R¹⁰ means methyl. In some cases, R¹⁰ means ethyl. In some cases, R¹⁰ is C1-3 alkyl. In some cases, R¹⁰ is C4-6 alkyl.

[0176] In Formula (V), R¹¹ is selected from hydrogen and C1-6 alkyl. In some cases, R¹¹ means hydrogen. In some cases, R¹¹ is C1-6 alkyl. In some cases, R¹¹ means methyl. In some cases, R¹¹ means ethyl. In some cases, R¹¹ is C1-3 alkyl. In some cases, R¹¹ is C4-6 alkyl.

[0177] In Formula (V), R¹² is selected from hydrogen and C1-6 alkyl. In some cases, R¹² means hydrogen. In some cases, R¹² is C1-6 alkyl. In some cases, R¹² means methyl. In some cases, R¹² means ethyl. In some cases, R¹² is C1-3 alkyl. In some cases, R¹² is C4-6 alkyl.

[0178] In Formula (V), R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo. In some cases, R¹³ means hydrogen. In some cases, R¹³ is C1-6 alkyl. In some cases, R¹³ means C6-20 aryl.

[0179] In some cases, R¹³ is unsubstituted C 1-6 alkyl. In some cases, R¹³ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R¹³ means - (CH2) mOH, where m is a number from one to 3. In some cases, R¹³ means -CH2OH. In some cases, R¹³ is C1-6 alkyl which is substituted with C1-6 alkoxy.

[0180] In some cases, R¹³ is C1-6 alkyl which is substituted with halo. In some cases, R¹³ is - (CH2) mX, wherein m is a number from one to three and X is halo. In some cases, R¹³ is C1-6 alkyl which is substituted with fluoro. In some cases, R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R¹³ means - (CH2) 2F.

[0181] In some cases, R¹³ means unsubstituted C6-20 aryl. In some cases, R¹³ means C6-20 aryl which is substituted with hydroxyl. In some cases, R¹³ means C6-20 aryl which is substituted with C1-6 alkoxy. In some cases, R¹³ means C6-20 aryl which is substituted with halo.

[0182] In some cases, R⁶, R⁷, R⁸and R⁹ means hydrogen. In some cases, R⁶, R⁷and R⁸ are hydrogen and R⁹ means halo. In some cases, R⁶, R⁷and R⁸ are hydrogen and R⁹ means fluoro.

[0183] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹⁰ means methyl. In some cases, R⁶, R⁷, R⁸ means hydrogen; R⁹ means halo; and R¹⁰ means methyl. In some cases, R⁶, R⁷, R⁸ means hydrogen; R⁹ means fluoro; and R¹⁰ means methyl.

[0184] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means hydrogen. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means hydrogen.

[0185] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means -CH2OH. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means -CH2OH. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means CH2OH.

[0186] In some cases, R⁶, R⁷, R⁸and R⁹ are hydrogen and R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means halo; and R¹³ means - (CH2) mF, wherein m is a number from one to

3. In some cases, R⁶, R⁷and R⁸ means hydrogen; R⁹ means fluoro; and R¹³ means (CH2) mF, where m is a number from one to three.

Formula (VI) [0187] The present disclosure provides a compound of Formula (VI):

<img file="PL2880035T3_D0026.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

Q is CR⁹ or N;

R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is selected from hydrogen and C1-6 alkyl; and

R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo; or a pharmaceutically acceptable salt thereof.

[0188] In Formula (VI), R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 16 haloalkyl. In some cases, R¹ means hydrogen. In some cases, R¹is C1-6 alkyl. In some cases, R¹ means methyl or ethyl. In some cases, R² means hydrogen. In some cases, R² is C1-6 alkyl. In some cases, R² means methyl or ethyl. In some cases, R¹ and R²means hydrogen.

[0189] In Formula (VI), R⁸ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁸ means hydrogen. In some cases, R⁸ means halo. In some cases, R⁸ means fluoro. In some cases, R⁸ means chloro. In some cases, R⁸ means bromo. In some cases, R⁸ is C1-6 alkyl. In some cases, R⁸means C1-6 haloalkyl. In some cases, R⁸ means C1-6 alkoxy. In some cases, R⁸ means C1-6 haloalkoxy. In some cases, R⁸ means hydroxyl. In some cases, R⁸ means cyano. In some cases, R⁸ means nitro.

[0190] In Formula (VI), Q is CR⁹ or N. In some cases, Q is CR⁹. In some cases, Q is N.

[0191] In Formula (VI), R⁹ is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro. In some cases, R⁹ means hydrogen. In some cases, R⁹ means halo. In some cases, R⁹ means fluoro. In some cases, R⁹ means chloro. In some cases, R⁹ means bromo. In some cases, R⁹ is C1-6 alkyl. In some cases, R⁹means C1-6 haloalkyl. In some cases, R⁹ means C1-6 alkoxy. In some cases, R⁹ means C1-6 haloalkoxy. In some cases, R⁹ means hydroxyl. In some cases, R⁹ means cyano. In some cases, R⁹ means nitro.

[0192] In Formula (VI), R¹⁰ is selected from hydrogen and C1-6 alkyl. In some cases, R¹⁰ means hydrogen. In some cases, R¹⁰ is C1-6 alkyl. In some cases, R¹⁰ means methyl. In some cases, R¹⁰ means ethyl. In some cases, R¹⁰ is C1-3 alkyl. In some cases, R¹⁰ is C4-6 alkyl.

[0193] In Formula (VI), R¹³ is selected from hydrogen, C1-6 alkyl, and C6-20 aryl, wherein each alkyl or aryl is unsubstituted or substituted by hydroxy, C1-6 alkoxy, or halo. In some cases, R¹³ means hydrogen. In some cases, R¹³ is C1-6 alkyl. In some cases, R¹³ means C6-20 aryl.

[0194] In some cases, R¹³ is unsubstituted C 1-6 alkyl. In some cases, R¹³ is C1-6 alkyl which is substituted with hydroxyl. In some cases, R¹³ means - (CH2) mOH, where m is a number from one to 3. In some cases, R¹³ means -CH2OH. In some cases, R¹³ is C1-6 alkyl which is substituted with C1-6 alkoxy.

[0195] In some cases, R¹³ is C1-6 alkyl which is substituted with halo. In some cases, R¹³ is - (CH2) mX, wherein m is a number from one to three and X is halo. In some cases, R¹³ is C1-6 alkyl which is substituted with fluoro. In some cases, R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R¹³ means - (CH2) 2F.

[0196] In some cases, R¹³ means unsubstituted C6-20 aryl. In some cases, R¹³ means C6-20 aryl which is substituted with hydroxyl. In some cases, R¹³ means C6-20 aryl which is substituted with C1-6 alkoxy. In some cases, R¹³ means C6-20 aryl which is substituted with halo.

[0197] In some cases, R⁸ and R⁹ means hydrogen. In some cases, R⁸is hydrogen and R⁹ means halo. In some cases, R⁸ is hydrogen and R⁹ means fluoro.

[0198] In some cases, R⁸ and R⁹ are hydrogen and R¹⁰ means methyl. In some cases, R⁸ is hydrogen; R⁹ means halo; and R¹⁰ means methyl. In some cases, R⁸ is hydrogen; R⁹ means fluoro; and R¹⁰ means methyl.

[0199] In some cases, R⁸ and R⁹ are hydrogen and R¹³ means hydrogen. In some cases, R⁸ is hydrogen; R⁹ means halo; and R¹³ means hydrogen. In some cases, R⁸ is hydrogen; R⁹ means fluoro; and R¹³ means hydrogen.

[0200] In some cases, R⁸ and R⁹ are hydrogen and R¹³ means -CH2OH. In some cases, R⁸ is hydrogen; R⁹ means halo; and R¹³ means -CH2OH. In some cases, R⁸ is hydrogen; R⁹ means fluoro; and R¹³ means -CH2OH.

[0201] In some cases, R⁸ and R⁹ are hydrogen and R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R⁸ is hydrogen; R⁹ means halo; and R¹³ means - (CH2) mF, where m is a number from one to 3. In some cases, R⁸ is hydrogen; R⁹ means fluoro; and R¹³ means - (CH2) mF, wherein m is a number from one to three.

Formula (VII) [0202] The present disclosure provides a compound of Formula (VII):

<img file="PL2880035T3_D0027.tif" />

wherein

R¹ and R² are independently selected from hydrogen, halo, C 1-6 alkyl, and C 1-6 haloalkyl;

R⁸ is selected from halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, hydroxy, cyano, and nitro;

R¹⁰ is C1-6 alkyl; and

R¹³ is hydrogen or C 1-6 alkyl;

or a pharmaceutically acceptable salt thereof.

[0203] In some embodiments of Formula (VII), R¹ and R² both are hydrogen. In other embodiments, R⁸ is halo, methyl, methoxy, or cyano. In still other embodiments, R⁸ means halo. In still other embodiments, R⁸means fluoro. In some embodiments, R¹⁰ means methyl, ethyl, or isopropyl. In other embodiments, R¹⁰ means methyl. In some embodiments, R¹³ means hydrogen, methyl, ethyl, or isopropyl. In other embodiments, R¹³ means hydrogen.

[0204] In certain embodiments of compounds of Formulas (I) - (VI), R⁶ and R⁷ can also mean methoxy, provided that neither R⁶ neither R⁷ does not mean methoxy when R¹⁰ means methyl. In other embodiments of Formulas (I) - (VII), the group

<img file="PL2880035T3_D0028.tif" />

is replaced by ^r15_nm ^^nh where is R for me?¹⁵ they are as defined here.

[0205] In some embodiments, the compound of the invention is a compound of Formula XX:

<img file="PL2880035T3_D0029.tif" />

wherein:

R^thirty and R³¹ each independently H or C1-4alkyl; R³² means H and R³³ means

<img file="PL2880035T3_D0030.tif" />

or R³² and R³³ taken together with the nitrogen to which they are attached, they form

<img file="PL2880035T3_D0031.tif" />

where R³⁶ is H, C 1-4 alkyl, or C 1-4 haloalkyl; and R³⁷ is H or C 1-4 alkyl;

Q is CH, CF, or N;

R³⁴ is H or methoxy;

R³⁵ is H, -CH2OH, or -CH2CH2F;

where R³⁴ is not methoxy when Q is CH, -NR³²R³³ means r³⁷-n^/ \ -I-R³⁷ means methyl; or a pharmaceutically acceptable salt thereof.

[0206] In some embodiments of Formula (XX), R^thirty and R³¹ both are H. In some embodiments, R³² means H and R³³ means

<img file="PL2880035T3_D0032.tif" />

In some embodiments, R³⁶ means -CH2CH2F. In other embodiments, R³² and R³³ taken together with the nitrogen to which they are attached, they form

<img file="PL2880035T3_D0033.tif" />

In still other embodiments, R³⁷ means methyl. In other embodiments, Q is N and R³⁴ means H. In yet other embodiments, Q is CH and R³⁴ means methoxy. In yet other embodiments, Q is CF and R³⁴means H. In other embodiments, R³⁵ means H.

[0207] In some embodiments, the present disclosure provides a compound selected from:

<td>Relationship</td><td>Structure</td><td>Chemical Name</td>

<td>1</td><td>0 oL ^^_nand_nXn H ^H</td><td>N- (3 - ((2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>2</td><td>/ ABOUT 'b .in</td><td>N- (3 - ((2 - ((6- (4-methylpiperazin-1-yl) pyridin-3-yl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>3</td><td>0 Λ Ίχν / ο H ^H</td><td>N- (3 - ((2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>4</td><td>/ ABOUT % 5&gt; b ° - ~ O</td><td>N- (3 - ((7- (hydroxymethyl) -2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>Relationship</td><td>Structure</td><td>Chemical Name</td>

<td>5</td><td>/ ABOUT "b ^p'° ur</td><td>N- (3 - ((7- (hydroxymethyl) -2 - ((6- (4-methylpiperazin-1-yl) pyridin-3-yl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy ) phenyl) acrylamide</td>

<td>6</td><td>about Λ ταΧ »^HHO</td><td>N- (3 - ((2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7- (hydroxymethyl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy ) phenyl) acrylamide; and</td>

<td>7</td><td>ABOUT ^ Y_NHΛ Αχ χΎ^H ? F</td><td>N- (3 - ((7- (2-fluoroethyl) -2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl ) acrylamide</td>

and its pharmaceutically acceptable salts.

[0208] In certain embodiments, the present disclosure provides a compound selected from:

<td>Relationship</td><td>Structure</td><td>Chemical Name</td>

<td>8</td><td>0 Yth 'n' '' Ύθ¹ ώγΟχτ H ^H</td><td>N- (3 - ((2 - ((1- (2- (dimethylamino) ethyl) -1H-indol-5-yl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>9</td><td>ABOUT Υ ^ ΝΗ Λ H ^H</td><td>N- (3 - ((2 - ((2 - ((dimethylamino) methyl) quinolin-6-yl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>Relationship</td><td>Structure</td><td>Chemical Name</td>

<td>10</td><td>X¹k5 ° n, T0 about /</td><td>N- (3 - ((5-cyclopropyl-2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>11</td><td>0 ==== ^ NH AND^{ν &lt;}\ γλ f ^ AAAj / H ^H</td><td>N- (3 - ((5-cyclopropyl-2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>12</td><td>0 NH "νΛ Q H ^H</td><td>N- (3 - ((2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -5- (pyrrolidin-1-yl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl ) acrylamide</td>

<td>13</td><td>0 NH blę, &lt;P ^^Νχχύ5 H ^H</td><td>N- (3 - ((2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -5- (pyrrolidin-1-yl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl ) oxy) phenyl) acrylamide</td>

<td>14</td><td>ABOUT nh fS OH "About x ^ Yol ^{N &lt;}SrA&gt; H ^H</td><td>N- (3 - ((5- (2-hydroxyethyl) -2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>15</td><td>ABOUT nh (fS ° H ^ Νχ'ΐΐ ^NFA / XA^?H ^H</td><td>N- (3 - ((2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -5- (2-hydroxyethyl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy ) phenyl) acrylamide</td>

<td>Relationship</td><td>Structure</td><td>Chemical Name</td>

<td>16</td><td>0 Λ 'N- | AND AO</td><td>N- (3 - ((5 - ((dimethylamino) methyl) -2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>17</td><td>0 === ^ NH AND^XWell ^^νΤΊ ^{ν &lt;?}Ηγ? F ^ Ά ΛΑ H ^H</td><td>N- (3 - ((5 - ((dimethylamino) methyl) -2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl ) oxy) phenyl) acrylamide</td>

<td>18</td><td>0 NH Λ YAAA^?H ^H</td><td>N- (3 - ((5- (dimethylamino) -2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>19</td><td>0 ^ = Y ^ NH λ ^ Ά " ^ⁿyo ⁿ ΑΆ Η ^H</td><td>N- (3 - ((5- (dimethylamino) -2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl ) acrylamide</td>

<td>20</td><td>0 Ν Η rS Ό Α&gt; r⁷^ Υλ αΜ&gt; Η ^Η</td><td>N- (3 - ((5- (2- (dimethylamino) ethyl) -2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) -acrylamide</td>

<td>21</td><td>0 rS Ό Α&gt; Υ ^^Νχι αΜ Η ^Η</td><td>N- (3 - ((5- (2- (dimethylamino) ethyl) -2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin -4ylo) oxy) phenyl) acrylamide</td>

<td>Relationship</td><td>Structure</td><td>Chemical Name</td>

<td>22</td><td>0 ^ 0 ABOUT /</td><td>N- (3 - ((5- (aziridin-1-yl) -2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl ) acrylamide</td>

<td>23</td><td>0 ^ "Άη Λ -NA and? ^ⁿya ⁿ^ VS&gt;_FAA_NAND_NAND/ H ^H</td><td>N- (3 - ((5- (aziridin-1-yl) -2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl ) oxy) phenyl) acrylamide</td>

<td>24</td><td>ABOUT^{sss /}^ NH -nA ^ 0 &amp; Α^Νγ ^&gt; ⁿ^ StA ΑΑ_να_νΛ / H ^H</td><td>N- (3 - ((5- (azetidin-1-yl) -2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl ) acrylamide</td>

<td>25</td><td>ABOUT NH '"A θ'? &amp; θ ^ ν ^ ΐι ^N | γΑ H ^H</td><td>N- (3 - ((5- (azetidin-1-yl) -2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl ) oxy) phenyl) acrylamide</td>

<td>26</td><td>^ΟΧΓ n 0 ^ 0 ABOUT /</td><td>N- (3 - ((2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -5- (piperidin-1-yl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl ) acrylamide</td>

<td>27</td><td>about Θ ^ ^ ΝΗ 'nA ^ 9 0 ^^νΎΑ ^N ιτΑ H ^H</td><td>N- (3 - ((2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -5- (piperidin-1-yl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl ) oxy) phenyl) acrylamide</td>

<td>Relationship</td><td>Structure</td><td>Chemical Name</td>

<td>28</td><td>0 ^ Ά_ΝΗAND^{n &lt;}Sta&gt; aAnTYa^H K</td><td>N- (3 - ((7-Cyclopropyl-2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>29</td><td>0 ^ N _NHά ^ⁿya ^ν^ ϊα^H K</td><td>N- (3 - ((7-Cyclopropyl-2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>thirty</td><td>0_NHAND N_about^ Yl JÓ3 "A</td><td>N- (3 - ((2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7- (methylsumonyl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>31</td><td>0 ^ = AA_HCL N_aboutYi Aa f ^^ naa^H Λ °</td><td>N- (3 - ((2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7- (methylsumonyl) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl ) acrylamide</td>

<td>32</td><td>0_NHAND AA_about^^Ν ΥΊ ⁿ^ StA&gt; AA_nand_nAND_n^?^H &lt;T</td><td>N- (3 - ((7-acetyl-2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>33</td><td>0 → A_NHAND A == A ^ Al Aa f ^^ naa</td><td>N- (3 - ((7-acetyl-2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide</td>

<td>Relationship</td><td>Structure</td><td>Chemical Name</td>

<td>34</td><td>about → A_NH</td><td>N- (3- (2- (4- (1- (2-fluoroethyl) azetidin-3-ylamino) -</td>

<td></td><td rowspan="2">oh₃^h</td><td>2-methoxy-phenylamino) -7H-pyrrolo [2,3-</td>

<td></td><td>d] pyrimidin-4-yloxy) phenyl) acrylamide</td>

<td>35</td><td>0 % Y_NH</td><td>N- (3- (2- (4- (1- (2-fluoroethyl) azetidin-3-ylamino) -</td>

<td></td><td rowspan="2">ά_H ^ NH oh₃^h</td><td>2-methoxy-phenylamino) -7H-pyrrolo [2,3-</td>

<td></td><td>d] pyrimidin-4-ylamino) phenyl) acrylamide; and</td>

<td>36</td><td>0 V ^ NH</td><td>N- (3 - ((2 - ((2- (piperidin-1-ylmethyl) -quinolin-6-</td>

<td></td><td rowspan="2">&amp; " OXCuto H ^H</td><td>yl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-</td>

<td></td><td>yl) oxy) phenyl) acrylamide</td>

and its pharmaceutically acceptable salts.

[0209] In certain embodiments, the present disclosure provides Compound 3, N- (3 ((2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2, 3-d] pyrimidin-4-yl) oxy) phenyl) acrylamide,

<img file="PL2880035T3_D0034.tif" />

and its pharmaceutically acceptable salts. In some embodiments, the present disclosure provides compound male salt of Compound 3, N- (3 - ((2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-] d] pyrimidin-4-yl) oxy) phenyl) acrylamide. In some embodiments, the present disclosure provides a hydrochloride salt of Compound 3, N- (3 ((2 - ((3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrole [2,3] d] pyrimidin-4-yl) oxy) phenyl) acrylamide.

[0210] The disclosed pharmaceutical compositions can be formulated in the form of a pharmaceutically acceptable salt of the disclosed compound. Pharmaceutically acceptable salts are non-toxic salts of the free base form of the compound that has the desired pharmacological activity of the free base. Such salts may be derived from inorganic or organic acids. Non-limiting examples of pharmaceutically acceptable salts include sulphates, metabisulfites, hydrogen sulphates, sulphites, bisulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, capronates, heptanes, propiolates, oxalates, malonates, succinates, suberiods, sebacates, fumarates, maleates, butyno-1,4-dionates, hexyno-1,6-dionates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulfonates, methylsulfonates, propylsulfonates, besylates, xylene sulphonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, phenylacetates, phenylpropionates, phenylbutyrate, citrates, lactates, γ-hydroxybutyrates, glycolates, winiany, and almonds. Lists of other suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pa., 1985. and almonds. Lists of other suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pa., 1985. and almonds. Lists of other suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pa., 1985.

Pharmaceutical compositions [0211] For therapeutic purposes, pharmaceutical compositions containing the compounds described herein may further comprise one or more pharmaceutically acceptable excipients. A pharmaceutically acceptable excipient is a non-toxic substance that is also biologically suitable for administration to a subject. Such excipients facilitate administration of the compounds described herein and are compatible with the active ingredient. Examples of pharmaceutically acceptable excipients include stabilizers, lubricants, surfactants, diluents, antioxidants, binders, dyes, fillers, emulsifiers, or flavor modifiers. In preferred embodiments, the pharmaceutical compositions of the embodiments are sterile compositions.

[0212] Sterile compositions are within the scope of the present disclosure, including compositions that are in compliance with the national and local regulations governing such compositions.

[0213] The pharmaceutical compositions and compounds described herein may be formulated as solutions, emulsions, suspensions or dispersions in suitable pharmaceutical solvents or vehicles, or as pellets, tablets, lozenges, suppositories, sachets, coated tablets, granules, powders, powders for reconstitution or capsule along with solid carriers according to conventional methods known in the art for the preparation of various dosage forms. The pharmaceutical compositions of the embodiments may be administered via a suitable route of administration, such as by oral, parenteral, rectal, nasal, topical or ocular routes or by inhalation. Preferably, the pharmaceutical compositions are formulated for intravenous or oral administration.

[0214] For oral administration, the compounds of the embodiments may be provided in a solid form, such as a tablet or capsule, or in the form of a solution, emulsion or suspension. For the preparation of oral compositions, the compounds of the embodiments may be formulated so as to obtain a dosage, e.g. from about 0.01 to about 50 mg / kg per day, or from about 0.05 to about 20 mg / kg per day, or from about 0.1 to about 10 mg / kg daily. Oral tablets may contain the active ingredient (s) mixed with compatible, pharmaceutically acceptable excipients such as diluents, disintegrants, binders, lubricants, sweeteners, flavoring agents, coloring agents and preservatives. Suitable inert fillers include sodium and calcium carbonate, sodium and calcium phosphate, lactose, starch, sugar, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol and the like. Exemplary liquid oral excipients include ethanol, glycerol, water and the like. Starch, polyvinyl pyrrolidone (PVP), sodium starch glycolate, microcrystalline cellulose and alginic acid are examples of disintegrants. Binders may include starch and gelatin. The lubricating agent, if present, may be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract or may be coated with an enteric coating. microcrystalline cellulose and alginic acid are examples of disintegrants. Binders may include starch and gelatin. The lubricating agent, if present, may be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract or may be coated with an enteric coating. microcrystalline cellulose and alginic acid are examples of disintegrants. Binders may include starch and gelatin. The lubricating agent, if present, may be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract or may be coated with an enteric coating.

[0215] Capsules for oral administration include hard and soft gelatin capsules. For the production of hard gelatine capsules, the active ingredient (s) may be mixed with a solid, semi-solid or liquid diluent. Soft gelatine capsules can be prepared by mixing the active ingredient with water, an oil such as peanut oil, olive oil, liquid paraffin, a mixture of short-chain fatty acid mono- and di-glycerides, with polyethylene glycol 400 or propylene glycol [ 0216] Liquids for oral administration may be in the form of suspensions, solutions, emulsions or syrups, or may be lyophilized or as a dry product for reconstitution with water or other suitable vehicle before use. Such liquid compositions may optionally contain: pharmaceutically acceptable excipients, as suspending agents (e.g., sorbitol, methylcellulose, sodium alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel and the like); non-aqueous vehicles, e.g. oil (e.g., almond oil or fractionated coconut oil), propylene glycol, ethyl alcohol or water; preservatives (e.g., methyl or propyl p-hydroxybenzoate or sorbic acid); wetting agents, such as lecithin; and, if necessary, flavoring or coloring agents. methyl or propyl p-hydroxybenzoate or sorbic acid); wetting agents, such as lecithin; and, if necessary, flavoring or coloring agents. methyl or propyl p-hydroxybenzoate or sorbic acid); wetting agents, such as lecithin; and, if necessary, flavoring or coloring agents.

[0217] The compositions may be formulated for rectal administration as a suppository. For parenteral use, including intravenous, intramuscular, intraperitoneal, intranasal, or subcutaneous administration, the compounds of the embodiments can be provided in sterile aqueous solutions or suspensions, buffered to an appropriate pH and isotonicity, or in a parenterally acceptable oil. Suitable aqueous vehicles include Ringer's solution and an isotonic sodium chloride solution. Such forms may be in unit dose forms, such as ampoules or single use injection devices, in multidose forms such as vials from which the appropriate dose may be administered, or in the form of a solid or pre-concentrate that may be used to prepare an injectable preparation. .

[0218] For nasal, inhalation, or oral administration, the pharmaceutical compositions can be administered using, for example, a spray formulation that also contains a suitable carrier.

[0219] For topical use, the compounds of the embodiments are preferably formulated in creams or ointments or in a similar medium suitable for topical administration. For topical administration, the compounds of the embodiments may be admixed with a pharmaceutical carrier at a concentration of about 0.1% to about 10% of drug to the medium. In another mode of administration, the compounds of the embodiments may use a formulation in the form of a patch to effect percutaneous delivery.

[0220] In certain embodiments, the present disclosure provides a pharmaceutical composition comprising a compound of formulas (I) - (VIII) and methylcellulose. In some embodiments, the methylcellulose is in a suspension of about 0.1, 0.2, 0.3, 0.4, or 0.5 to about 1%. In some embodiments, the methylcellulose is in a suspension in an amount of about 0.1 to about 0.5, 0.6, 0.7, 0.8, 0.9, or 1%. In some embodiments, methylcellulose is in suspension at about 0.1 to about 1%. In some embodiments, the methylcellulose is suspended in an amount of about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.8, or 1%. In some embodiments, methylcellulose is in suspension at about 0.5%.

[0221] As used herein, the terms "treat" or "treatment" include both "preventive" and "curative" treatments. "Preventive" treatment is intended to postpone the development of the disease, symptoms of the disease or medical condition, suppression of symptoms that may arise or reduce the risk of developing or recurring the disease or symptom. "Curative" treatment includes reducing the severity or suppressing the deterioration of an existing disease, symptom or condition. Thus, the treatment includes alleviating or preventing deterioration of existing symptoms of the disease, preventing the onset of additional symptoms, ameliorating the disease state or preventing the systemic causes of symptoms, inhibiting the disorder or disease, e.g. arresting the development of the disorder or disease, alleviating the disorder or disease, causing regression of the disease, or disorder, relieving the condition caused by the disease or disorder, or arresting the symptoms of the disease or disorder. [0222] One of skill in the art can modify formulations within the scope of the disclosure herein to provide numerous formulations for a particular route of administration. In particular, these compounds can be modified to make them more soluble in water or other substrate. It is also well known to those skilled in the art of modifying the route of administration and the dosage regimen of a particular compound to obtain the pharmacokinetic properties of the present compounds for maximum beneficial effects on the patient. [0222] One of skill in the art can modify formulations within the scope of the disclosure herein to provide numerous formulations for a particular route of administration. In particular, these compounds can be modified to make them more soluble in water or other substrate. It is also well known to those skilled in the art of modifying the route of administration and the dosage regimen of a particular compound to obtain the pharmacokinetic properties of the present compounds for maximum beneficial effects on the patient. [0222] One of skill in the art can modify formulations within the scope of the disclosure herein to provide numerous formulations for a particular route of administration. In particular, these compounds can be modified to make them more soluble in water or other substrate. It is also well known to those skilled in the art of modifying the route of administration and the dosage regimen of a particular compound to obtain the pharmacokinetic properties of the present compounds for maximum beneficial effects on the patient.

[0223] The term "subject" refers to a mammal as a subject in need of treatment such as a human.

[0224] The compounds may be administered to a subject in need of treatment for a proliferative disorder. An example of a proliferative disorder is cancer. In some cases, the compounds are useful for the treatment of sarcoma, epidermoid tumor, fibrosarcoma, cervical tumor, gastric cancer, skin cancer, leukemia, lymphoma, lung cancer, non-small cell lung cancer, colon cancer, CNS tumor, melanoma, ovarian cancer, cancer kidney, prostate cancer, breast cancer, liver cancer, head and neck cancer and pancreatic cancer.

[0225] The compounds may also be administered to a subject in need of treatment for a disease or condition mediated by a protein kinase selected from the group consisting of EGFR, EGFR (T790M), BLK, BMX / ETK, BTK, JAK1, JAK2, JAK3, TEC, TXK, FLT3, and FLT3 (D835Y). In some cases, these compounds are useful for the treatment of tumors, tumors, inflammatory diseases, autoimmune diseases, or immunosuppressed diseases. In other embodiments, such diseases are mediated by at least one kinase selected from BTK, JAK3, ITK, and BMX. In other embodiments, tumors, tumors, inflammatory diseases, autoimmune diseases or immunosuppressed diseases mediate malfunctioning B lymphocytes, T lymphocytes, or both. In other embodiments, inflammatory diseases, stomach cancer, skin cancer, leukemia, lymphoma, lung cancer, non-small cell lung cancer, colon cancer, CNS tumor, melanoma, ovarian cancer, kidney cancer, prostate cancer, breast cancer, liver cancer, head and neck cancer, or pancreatic cancer. In other embodiments, the diseases are autoimmune diseases or graft-induced inflammatory disorders including, but not limited to, allograft, graft versus host disease, or autoimmune diabetes.

[0226] In one aspect, the compounds and pharmaceutical compositions of the embodiments can be administered to a subject in need of treatment of a condition associated with EGFR-inhibitory activity that is directed essentially to mutant EGFR but substantially not of wild-type EGFR. In some embodiments, the mutant EGFR contains the T790M mutation. The present disclosure provides the use of a compound of Formulas (I) - (VIII) for the manufacture of a medicament for the treatment of such conditions, and the use of these compounds and salts in the treatment of such conditions. In another aspect, the compounds and pharmaceutical compositions may be administered to a subject in need of treatment of a condition associated with FLT3 inhibitory activity directed to substantially mutated FLT3, but not substantially wild FLT3. In some embodiments, the mutant FLT3 contains the D835Y mutation.

[0227] In another aspect, the present disclosure provides a method of inhibiting mutant EGFR in a cell, comprising contacting the cell with an effective amount of at least one compound of Formulas (I) - (VIII) or a salt thereof, and / or at least one pharmaceutical composition according to examples wherein the contacting is in vitro, ex vivo, or in vivo. In some embodiments, the mutant EGFR contains the T790M mutation. In another aspect, the present disclosure provides a method of inhibiting mutant FLT3 in a cell, comprising contacting the cell with an effective amount of at least one compound of Formulas (I) - (VIII) or a salt thereof, and / or at least one pharmaceutical composition containing said compound or salt wherein the in vitro, ex vivo or in vivo contacting is performed.

[0228] In the inhibiting methods of the embodiments, the term "effective amount" means an amount sufficient to inhibit a target receptor, e.g. EGFR mutant, but not wild type EGFR, or mutant FLT3, but not wild type FLT3. In some embodiments, the mutant EGFR contains the T790M mutation. In other embodiments, the mutant FLT3 contains the D835Y mutation. The degree of inhibition can be measured using typical analytical methods such as those described below. Such modulation is useful in a variety of contexts, including in vitro assays. Other settings include ex vivo and in vivo.

[0229] In treatment methods, according to embodiments, "effective amount" means an amount or dose sufficient to generally achieve the desired therapeutic benefit in subjects in need of such treatment. Effective amounts or doses of the compounds of the embodiments may be determined by routine methods such as modeling, dose escalation or clinical testing with routine factors, e.g., mode or route or administration of drug delivery, pharmacokinetics of the agent, severity and course of infection, health status individual, condition and weight and evaluation of the attending physician. An exemplary dose ranges from about 1 μg to 2 mg active agent per kilogram body weight of the subject per day, preferably about 0.05 to 100 mg / kg / day, or about 1 to 35 mg / kg / day, or about 0.1 to 10 mg / kg / day. The total dosage can be given in single or divided dosage units (e.g., BID, TID, QID). [0230] Once the disease of the patient has improved, the dose may be adjusted for preventive or maintenance treatment. For example, the dosage or frequency of administration, or both, may be reduced depending on the symptoms to a level at which the desired therapeutic effect or prophylactic effect is maintained. Of course, if the symptoms have been alleviated to an appropriate level, treatment may stop. Patients may, however, require treatment with interruptions over a longer period of time depending on the relapse of symptoms. Patients may require long-term treatment. Drug combination [0231] The methods of the embodiments include administering an effective amount of at least one compound of Formula (I) - (VIII) or their embodiments; optionally, the compound may be administered in combination with one or more additional therapeutic agents, especially therapeutic agents that are known to be useful in the treatment of a proliferative disease or cancer that affects the subject. In some embodiments, the one or more therapeutic agents are selected from antitumor agents (such as cell signal transduction inhibitors, mitosis inhibitors, alkylating agents, antimetabolites, intercalating anti-cancer drugs, topoisomerase inhibitors, immunotherapeutic agents or antihormonal agents), steroid drugs, methotrexate, leflunomide,

[0232] Additional active ingredients may be administered in a separate pharmaceutical composition from a compound of the embodiments or may be included with the compound of the embodiments in a single pharmaceutical composition. Additional active ingredients may be administered concurrently with, before or after administration of the compound of the embodiments.

Chemical Synthesis [0233] Exemplary chemical compounds useful in the methods of the embodiments will now be described by reference to pictorial synthetic schemes for their general preparation below and specific embodiments that are even further. It will be appreciated by skilled artisans that in order to obtain various compounds herein, the starting materials may be selected such that finally the desired substituents will be obtained by reaction schemes with or without protecting, as the case may be, to produce the desired product. Alternatively, it may be necessary or desirable to use, instead of a definitively desired substituent, a suitable group that can be usedcarry out the reaction scheme and substitute for the desired substituent in a suitable manner. In addition, one skilled in the art will recognize that the transformations shown in the following schemes below can be performed in any order that is consistent with the functionality of the individual side groups. Each of the reactions illustrated in the general schemes is preferably carried out at a temperature from about 0 ° C to the boiling point of the organic solvent used. Unless otherwise specified, the variables are as defined above in relation to Formula (I). One of skill in the art will also appreciate that the methods described in these exemplary schemes may also be used to prepare compounds of Formula (VIII), and compounds of Formulas (II) - (VII).

[0234] A representative synthesis for the subject compounds is shown in the Scheme

<img file="PL2880035T3_D0035.tif" />

[0235] In Scheme 1, variables are as defined herein. As discussed below, X^2a and X^2b contain a leaving group. The starting materials can be obtained from commercial sources or through well-established synthetic procedures.

[0236] With reference to Scheme 1, the reaction of Compound 1-A with Compound 1-B by nucleophilic reaction forms Compound 1-C. In Compound 1-A, a hydroxyl group is a nucleophile that can provide an ether bond in Compound 1-C. The nucleophile may be reacted in a nucleophilic substitution reaction in which the nucleophile replaces the leaving group on the second reagent. In alternative embodiments, analogs of aniline or thiophenol 1-A are used to provide compounds in which X¹ is NH or S. In Compound 1-B, X^2acontains a leaving group. Examples of the leaving group include, but are not limited to, halo, triflate, fluorosulfonate, tosylate or mesylate.

[0237] Still referring to Scheme 1, the reaction of Compound 1-C with Compound 1-D under the Buchwald-Hartwig cross-coupling reaction conditions provides Compound 1-E. In the 1-D compound, the amino group is a nucleophile that can provide amine linkage in Compound 1-E. The nucleophile may react in a nucleophilic aromatic substitution in which the nucleophile replaces the leaving group on the second reagent. In Union 1-C, X^2b contains a leaving group. Examples of the leaving group include, but are not limited to, halo, triflate, fluorosulfonate, tosylate or mesylate.

[0238] Still referring to Scheme 1, the nitro group in Compound 1-E is reduced to give the amino group in Compound 1-F. The reduction of the nitro group can be carried out using an acid catalyst and a metal, or with a metal catalyst under a hydrogen gas atmosphere. In the reaction with an acid catalyst, metal, zinc, lithium, sodium or tin (typically tin chloride) may be used as the metal, and inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid; organic carboxylic acids such as acetic acid or trifluoroacetic acid; amine salts with acids, such as ammonium chloride, can be used as an acid catalyst. Furthermore, in the reduction process using a metal catalyst in a hydrogen gas atmosphere, palladium, nickel, platinum, ruthenium or rhodium may be used as the metal catalyst.

[0239] Still referring to Scheme 1, amidation of Compound 1-F gives compound of Formula (I). In the amidation reaction, Compound 1-F is reacted with an acryloyl derivative containing a leaving group. Examples of the leaving group include, but are not limited to, halo, triflate, fluorosulfonate, tosylate or mesylate. The amidation reaction may be carried out in a solvent such as dimethylformamide or dichloromethane in the presence of a base such as triethylamine or diisopropylethylamine. The amidation reaction can be carried out using a coupling agent such as, for example, dicyclohexylcarbodiimide (DCC), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC), or N- [dimethylamino-1H-1,2,3-triazolo [4, 5-b] pyridin-1-ylmethylene] -N-methyl-methanamine (HATU) together with 1-hydroxy-1H-benzotriazole (HOBT).

[0240] In some embodiments, a representative synthesis for the subject compounds is provided in Scheme 2.

<img file="PL2880035T3_D0036.tif" />

Compound 3 [0241] In some embodiments, a representative synthesis for the subject compounds is shown in Scheme 3.

Scheme 3

O ΧΚο, O jfO

WO? Aa Jίο A

Η '- "Λ.-' μ" N_cm 1 ^ 00 ,,,. &lt;«, Balloon N;

FenMH₄C!

DIPEA. DCM about »'o

Compound 36

[0242] In some embodiments, a representative synthesis for the subject compounds is shown in Scheme 4.

<img file="PL2880035T3_D0037.tif" />

[0243] In some embodiments, a representative synthesis for the subject compounds is shown in Scheme 5.

Scheme b

CiĄOAcL tir-τ'.ιΐϊ it ..&gt; 38h

KjiOOj

SC-9C fl.YF, -U PtIYS-PIŁOK t-BdOH / KjCO 3

Fe / NH.Cl

DIPEA, DCM

Compound 29 [0244] Compounds in which X¹ means NH is prepared according to Scheme 5-1, as illustrated by the example of Compound 35.

<img file="PL2880035T3_D0038.tif" />

[0245] Accordingly, and as described in more detail herein, the present disclosure provides a method of preparing a compound of the present disclosure, said method comprising: reacting a compound of formula

<img file="PL2880035T3_D0039.tif" />

thereby producing a compound of the formula

<img file="PL2880035T3_D0040.tif" />

where R³, R⁶, R⁷, R⁸ R¹⁰, R¹¹, R¹², R¹³, Q, in are as defined herein and X² means a leaving group.

[0246] Accordingly, and as described in more detail herein, the present disclosure provides a method for preparing a compound of the present disclosure, said method comprising: reacting a compound of formula

<img file="PL2880035T3_D0041.tif" />

thereby producing a compound of the formula

<img file="PL2880035T3_D0042.tif" />

where X¹, R³, R⁶, R⁷, R⁸, R¹⁰, R¹¹, R¹², R¹³, R¹⁸, R¹⁹, Q, in are as defined herein and X² means a leaving group.

[0247] Accordingly, and as described in more detail herein, the present disclosure provides a method for producing a compound of the present disclosure, said method comprising:

reducing the nitro group of a compound of the formula

<img file="PL2880035T3_D0043.tif" />

conducting the amidation reaction with an acryloyl derivative containing a leaving group;

thus giving a compound of Formula (I).

[0248] Accordingly, and as described in more detail herein, the present disclosure provides a method for producing a compound of the present disclosure, said method comprising:

reducing the nitro group of a compound of the formula

<img file="PL2880035T3_D0044.tif" />

conducting the amidation reaction with an acryloyl derivative containing a leaving group;

thus giving a compound of Formula (VIII).

[0249] In some cases, the above methods further comprise the salt formation step of the compound of the present disclosure. Embodiments are devoted to other methods described herein; and a product made by any of the methods described herein.

Examples [0250] The following examples are given to illustrate and not to limit the invention.

Example 1: Synthesis of Compounds 1 and 4:

[0251]

<img file="PL2880035T3_D0045.tif" />

[0252] Synthesis of N- (3 - ((7- (hydroxymethyl) -2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) 7H-pyrrolo [2,3-d] pyrimidin-4-one ylo) oxy) phenyl) acrylamide (Compound 4) and N- (3 - ((2 - ((4- (4-methylpiperazin-1-yl) phenyl) amino) -7H-pyrrolo [2,3-d] pyrimidin-4-yl ) oxy) phenyl) acrylamide (Compound 1) and their intermediates is shown in Scheme 6 and described below.

Synthesis_2,4-dichloro-7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d] pyrimidine (Compound 6-B):

<img file="PL2880035T3_D0046.tif" />

[0254] Sodium hydride (60%, 46.7mg, 3.06 mmol) was added to a mixture of Compound 2-A (575mg, 3.06mmol) and 2- (trimethylsilyl) ethoxymethyl chloride (561mg, 3.37mmol) in tetrahydrofuran (5mL) in 0 ° C with stirring. The reaction mixture was allowed to warm to room temperature and stirred for 3 hours, then quenched with water (5 mL). The mixture was extracted with ethyl acetate (10 mL x3). The organic layers were combined, washed with brine, dried over Na 2 SO 4 and filtered. The filtrate was concentrated, and the crude material was purified by column chromatography (PE / EA = 20/1) to give Compound 6-B (520 mg, 53.4% ​​yield, M + H⁺= 319.27) as a light yellow solid.

Synthesis of 2-chloro-4- (3-nitrophenoxy) -7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3d] pyrimidine (Compound 6-C):

[0255] νο₂

<img file="PL2880035T3_D0047.tif" />

SEM [0256] To a mixture of Compound 6-B (200 mg, 0.628 mmol) and 3-nitrophenol (96.2 g, 0.691 mmol) in dimethylformamide (2 mL) was added K2CO3 (173.7 mg, 1.26 mmol). The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was then filtered. The filtrate was diluted with water, then extracted with ethyl acetate. The organic layer was washed with water, brine, and dried over Na2SO4. After filtration and volatiles removal in vacuo, the crude product was purified by flash column chromatography (PE / EA = 20/1) to give Compound 6-C (200 mg, 75.6% yield, M + H⁺= 421.92) in the form of a white solid.

Synteza_N- (4- (4-methylpiperazin-1-yl) phenyl) -4- (3-nitrophenoxy) -7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d] pyrimidin- 2-amines (Compound 6-D):

[0257] no₂

SEM [0258] A mixture of Compound 6-C (150 mg, 0.356 mmol), 4- (4-methylpiperazino) aniline (70 mg, 0.356 mmol), tris (dibenzylideneacetone) dipalladium (36 mg, 0.0356 mmol), dicyclohexyl (2 ') , 4 ', 6'-triisopropylbiphenyl-2-yl) phosphine (100 mg, 0.214 mmol) and potassium carbonate (197 mg, 1.424 mmol) in t-butanol (8 mL) was stirred under argon at 80 ° C overnight. After cooling to room temperature, the reaction mixture was filtered through a pad of celite. The celite layer was washed with methanol and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (DCM / MeOH = 20/1) to give Compound 6-D (180 mg, M + H).⁺ = 576.23) as a yellow solid.

Synthesis 4- (3-aminophenoxy) -N- (4- (4-methylpiperazin-1-yl) phenyl) -7 - ((2 (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d] pyrimidine 2-amines (Compound 6-E):

[0259]

<img file="PL2880035T3_D0048.tif" />

[0260] Compound 6-D (180mg, 0.312 mmol) was dissolved in ethanol (6 mL) and water (2 mL) was added. Then iron powder (90 mg, 1.61 mmol) and iron chloride (230 mg, 4.3 mmol) were added, and the resulting mixture was heated under reflux for 3 hours. The reaction mixture was cooled to room temperature and filtered through a pad of celite. The ethanol was removed in vacuo, and the obtained residue was basified with sodium bicarbonate and extracted with ethyl acetate. The organic layer was separated and dried with anhydrous sodium sulfate, concentrated, and purified by flash chromatography with 20: 1 dichloromethane-methanol to give Compound 6-E (170 mg, M + H).⁺ = 546) in the form of a white solid.

Synteza_N- (3- (2- (4- (4-methylpiperazin-1-yl) phenylamino) -7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d] pyrimidin-4 -yloxy) phenyl) acrylamide (Compound 6-F)

<img file="PL2880035T3_D0049.tif" />

[0262] Acryloyl chloride (33.8 mg, 0.374 mmol) was added dropwise to a solution of Compound 6-E (170 mg, 0.312 mmol) and diisopropylethylamine (55 mg, 0.426 mmol) in methylene chloride (3 mL) at 0 ° C. The reaction mixture was stirred for 1 hour. Water was added to quench the reaction. The organic layer was washed with water, brine, and dried over Na2SO4. After filtration, removal of volatiles was carried out in vacuo. The crude product was purified by chromatography (DCM / MeOH = 20/1) to give Compound 6-F (125 mg, 66.9% yield, M + H⁺ = 600.8) in the form of a white solid.

Synthesis of N- (3- (7- (hydroxymethyl) -2- (4- (4-methylpiperazin-1-yl) phenylamino) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy) phenyl) acrylamide (Compound 4) [0263]

<img file="PL2880035T3_D0050.tif" />

[0264] Compound 6-F (125 mg, 0.208 mmol) in methylene chloride (3 mL) and trifluoroacetic acid (1 mL) were stirred at room temperature for 3 hours. Monitoring by thin layer chromatography showed that all starting material had been consumed. Saturated aqueous NaHCO 3 was then added to the reaction mixture at 0 ° C. The reaction mixture was extracted with methylene chloride. The organic layer was washed with water, brine, dried over Na 2 SO 4 and filtered. The filtrate was concentrated and the crude material was purified by column chromatography (DCM / MeOH = 20/1) to give Compound 4 (70 mg, 71.5% yield, M + H⁺= 500.5) in the form of a white solid.

Synthesis of N- (3- (2- (4- (4-methylpiperazin-1-yl) phenylammo) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy) phenyl) acrylamide (Compound 1)

<img file="PL2880035T3_D0051.tif" />

[0266] A solution of Compound 4 (100 mg, 0.2 mmol) in methanol (2 mL) was saturated with ammonia. The reaction mixture was stirred overnight at room temperature. Monitoring with LC-MS showed that all of the starting material was consumed. The solvent was concentrated and the crude material was purified by column chromatography (DCM / MeOH = 20/1) to give Compound 1 (60 mg, 63.8% yield, M + H⁺= 470.5) as a light yellow solid.

Example 2: Synthesis of Compounds 2 and 5:

[0267]

<img file="PL2880035T3_D0052.tif" />

[0268] Synthesis of N- (3- (7- (hydroxymethyl) -2- (6- (4-methylpiperazin-1-yl) pyridin-3-ylamino) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy ) phenyl) acrylamide (Compound 5) and N- (3- (2- (6 (4-methylpiperazin-1-yl) pyridin-3-ylamino) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy) phenyl ) acrylamide (Compound 2) and their intermediates is shown in Scheme 7 and described below.

Synteza_N- (6- (4-methylpiperazin-1-yl) pylidin-3-yl) -4- (3-nitrophenoxy) -7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3- d] pyrimidin-2-amino (Compound 7-B):

<img file="PL2880035T3_D0053.tif" />

[0270] Compound 7-B (62% yield from 3, M + H⁺= 577.3) was prepared according to the procedure of Compound 6-D using 3-amino-6- (4-methyl-1-piperazinyl) pyridin hydrochloride instead of 4- (4-methylpiperazno) aniline.

Synteza_4- (3-Amino-phenoxy) -N- (6- (4-methylpiperazin-1-yl) pyridin-3-yl) -7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3- d] pyrimidin-2-amino (Compound 7-C):

[0271]

<img file="PL2880035T3_D0054.tif" />

[0272] Compound 7-C (80% yield from Compound 7-B, M + H⁺= 547.3) was prepared according to the procedure of Compound 6-E.

Synthesis of NOP-phenyl-methyl-piperazin-1-yl-pyridin-1-yl-hemopyr-1-dimethyl-1-ylthiocloxy-ethyl-1H-pyrrolo-4-pyrimidine-3-yloxyphenyl-amide (Compound 7-D)

<img file="PL2880035T3_D0055.tif" />

[0274] Compound 7-D (67% yield from Compound 7-C, M + H⁺= 601.3) was prepared according to the procedure of Compound 6-F.

Synthesis of N- (3- (7- (hydroxymethyl) -2- (6- (4-methylpiperazin-1-yl) pyridin-3-ylamino) -7H-pyrrolo-3-yl} irimidine ^ -yloxyphenylacrylamide (Compound 5):

<img file="PL2880035T3_D0056.tif" />

[0276] Compound 7-E (70% yield from Compound 5, M + H⁺= 501.6) was prepared according to the procedure of Compound 4.

Synthesis of N- (3 - (2- (6- (4-methylpiperazin-1-yl) pyridin-3-ylamino) -7H-pyrrolo [2,3-d] irimidine ^ -yloxyphenyl] amide (Compound 2):

[0277]

<img file="PL2880035T3_D0057.tif" />

[0278] Compound 2 (62% yield from Compound 5, M + H⁺= 471.5) was prepared according to the procedure of Compound 1.

Example 3: Synthesis of Compounds 3 and 6:

[0279]

<img file="PL2880035T3_D0058.tif" />

Synthesis of N- (3- (2- (3-fluoro-4- (4-methylpiperazin-1-yl) -phenylamino) -7 (hydroxymethyl) -7H-pyrrolo [2,3-d] pyrimidin-4-one ethoxy) phenyl) acrylamide (Compound 6) and N (3- (2- (3-fluoro-4- (4-methylpiperazin-1-yl) phenylamino) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy) phenyl) acrylamide (Compound 3) and their intermediates is shown in Scheme 8 and described below.

Synthesis of N- (3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) -4- (3-nitrophenoxy) -7 - ((2 (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3- d1-pyrimidin-2-amine (Compound 8-B) [0281]

<img file="PL2880035T3_D0059.tif" />

[0282] Compound 8-B (% yield from Compound 8-A, M + H⁺= 594.3) was prepared according to the procedure of Compound 6-D using 3-fluoro-4- (4-methylpiperazin-1-yl) aniline in place of 4- (4-methylpiperazine) aniline.

Synteza_4- (3-aminophenoxy) -N- (3-fluoro-4- (4-methylpiperazin-1-yl) phenyl) -7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3- d] pyrimidin-2-amine (Compound 8-C):

<img file="PL2880035T3_D0060.tif" />

[0284] Compound 8-C (85% yield from Compound 8-B, M + H⁺= 564.3) was prepared according to the procedure of Compound 6-E.

Synteza_N- (3- (2- (3-fluoro-4- (4-methylpiperazin-1-yl) phenylamino) -7 - ((2- (trimethylsilyl) ethoxy) methyl) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy) phenyl) acrylamide (Compound 8-D)

<img file="PL2880035T3_D0061.tif" />

[0286] Compound 8-D (75% yield from Compound 8-C, M + H⁺= 618.3) was prepared according to the procedure of Compound 6-F.

Synthesis of N- (3- (2- (3-fluoro-4- (4-methylpiperazin-1-yl) -phenylamino) -7- (hydroxymethyl) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy) phenyl) acrylamide (Compound 6):

[0287]

<img file="PL2880035T3_D0062.tif" />

[0288] Compound 6 (78% yield from Compound 8-D, M + H⁺= 518.6) was prepared according to the procedure of Compound 4.

Synthesis of N- (3- (2- (3-fluoro-4- (4-methylpiperazin-1-yl) -phenylamino) -7H-pyrrolo [2,3d] pyrimidin-4-yloxy) phenyl) acrylamide (Compound 3):

<img file="PL2880035T3_D0063.tif" />

[0290] Compound 3 (83% yield from Compound 6, M + H⁺= 488.5) was prepared according to the procedure of Compound 1.

Example 4: N- (3 - (7- (2-fluoroethyl) -2- (4- (4-methylpiperazin-1-yl) phenylamino) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy) phenyl) acrylamide (Compound 7) [0291]

<img file="PL2880035T3_D0064.tif" />

[0292] Synthesis of N- (3- (7- (2-fluoro-ethyl) -2- (4- (4-methylpiperazin-1-yl) -phenylamino) -7H-pyrrolo [2,3-dlpyrimidin-4-yloxy) phenyl) acrylamide (Compound 7) and its intermediates are shown in Scheme 9 and described below.

Synthesis of 2,4-dichloro-7- (2-fluoro-ethyl) -7H-pyrrolo [2,3-d] pyrimidine (Compound 9-B)

<img file="PL2880035T3_D0065.tif" />

[0294] Sodium hydride (60%, 424 mg, 10.6 mmol) was added to a mixture of Compound 9-A (1g, 5.3 mmol) and BrCH2CH2F (1.519 g, 11.9 mmol) in acetonitrile (10 mL) at room temperature. The reaction mixture was stirred for 4 hours, then the reaction was quenched with water and then extracted with ethyl acetate. The organic layer was washed with brine, dried over Na 2 SO 4 and filtered. The filtrate was concentrated and the crude material was purified by column chromatography (PE / EA = 20/1) to give Compound 9-B (1.1 g, 90% yield, M + H⁺= 234.0) as a light yellow solid.

Synthesis of 2-chloro-7- (2-fluoroethyl) -4- (3-nitrophenoxy) -7H-pyrrolo [2,3-d] pyrimidine (Compound

9-C)

<img file="PL2880035T3_D0066.tif" />

[0296] Compound 9-C (82% yield from Compound 9-B, M + H⁺= 337.0) was prepared according to the procedure of Compound 6-C.

Synthesis of 7- (2-fluoroethyl) -N- (4- (4-methylpiperazin-1-yl) phenyl) -4- (3-nitrophenoxy) -7H-pyrrolo [2,3-d] pyrimidin-2-amine (Compound 9) -D)

<img file="PL2880035T3_D0067.tif" />

[0298] Compound 9-D (73% yield from Compound 9-C, M + H⁺= 492.2) was prepared according to the procedure of Compound 6-D.

Synthesis of 4- (3-aminophenoxy) -7- (2-fluoroethyl) -N- (4- (4-methylpiperazin-1-yl) phenyl) -7H-pyrrolo [2,3-d] pyrimidin-2-amine (Compound 9) -E)

<img file="PL2880035T3_D0068.tif" />

[0300] Compound 9-E (81% yield from Compound 9-D, M + H⁺= 462.2) was created according to the procedure of Compound 6-E.

Synthesis of N- (3- (7- (2-fluoroethyl) -2- (4- (4-methylpiperazin-1-yl) phenylamino) -7H-pyrrolo [2,3d] pyrimidin-4-yloxy) phenyl) acrylamide ( Compound 7)

<img file="PL2880035T3_D0069.tif" />

[0302] Compound 7 (77% yield from Compound 9-E, M + H⁺= 516.6) was formed according to the procedure of Compound 6-F.

Example 25_Synthesis of N- (3- (2- (4- (1- (2-fluoroethyl) azetidin-3-ylamino) -2-methoxyphenylamino) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy) phenyl) acrylamide (Compound 34)

<img file="PL2880035T3_D0070.tif" />

Synthesis of tert-butyl 3- (3-methoxy-4-nitrophenylamino) azetidine-1-carboxylate [0304]

<img file="PL2880035T3_D0071.tif" />

[0305] To a 100 mL 3-neck round bottom flask equipped with a reflux condenser, 4-fluoro-2-methoxy-1-nitrobenzene (4.086 g) and tert-butyl 3-aminoazetidine-1-carboxylate (4.4 g), triethylamine (9.6) were added. mL), and dimethyl sulfoxide (20 mL). The reaction mixture was heated at 95 ° C for 8 hours. The reaction mixture was poured into water (200 mL) and extracted with ethyl acetate (50 mL x3). The organic layer was washed with brine (50 mL x2), dried over sodium sulfate, and concentrated completely under reduced pressure at 40 ° C to give the title compound (9g) which was used without further purification.

Synthesis of N- (3-methoxy-4-nitrophenyl) azetidin-3-amine [0306]

<img file="PL2880035T3_D0072.tif" />

[0307] To T-butyl 3- (3-methoxy-4-nitrophenylamino) azetidine-1-carboxylate (9 g), TFA (18 mL) was added at room temperature. The reaction mixture was stirred for 15 min at room temperature and then concentrated under reduced pressure at 40 ° C to give the title compound as a TFA salt (7.24 g).

Synthesis of 1- (2-fluoro-ethyl) -N- (3-methoxy-4-nitrophenyl) azetidin-3-amine

<img file="PL2880035T3_D0073.tif" />

[0309] To N- (3-methoxy-4-nitrophenyl) azetidin-3-amine (3 g) Cs 2 CO 3 (12 g) was added in 1,2-bromofluoroethane (1.5 g) in DMF (30 mL). The reaction mixture was heated at 50 ° C for 8 h. The reaction mixture was poured into water and extracted in ethyl acetate (100 mL x3). The organic layer was washed with brine (100 mL x2), dried over sodium sulfate, and concentrated under reduced pressure. The crude material was purified by column chromatography (DCM / MeOH = 50/1 as eluant) to give the title compound (1.35 g, 51% yield over 3 steps) as a yellow solid.

Synthesis of N1 - (1- (2-fluoro-ethyl) -azetidin-3-yl) -3-methoxybenzene-1,4-diamine [0310]

<img file="PL2880035T3_D0074.tif" />

A solution of 1- (2-fluoro-ethyl) -N- (3-methoxy-4-nitrophenyl) azetidin-3-amine (2.6 g) and Pd / C (1 g) in 1,4-dioxane (50 mL) subjected to hydrogenation for 4 hours at room temperature. The reaction mixture was filtered through diatomaceous earth, washing with MeOH. The filtrate was concentrated and purified by column chromatography (DCM / MeOH = 50/1 as eluent) to provide the title compound (1.57g, 68% yield, M + H).⁺ = 240.2).

Pivalate (2- (4- (1- (2-fluoro-ethyl) -azetidin-3-ylamino) -2-methoxyphenylamino) -4- (3-nitrophenoxy) -7H-pyrrolo [2,3-d] pyrimidin-7-yl synthesis ) methyl (Compound 34-A)

<img file="PL2880035T3_D0075.tif" />

A mixture of N1- (1- (2-fluoro-ethyl) -azetidin-3-yl) -3-methoxybenzene-1,4-diamine (870 mg, 3.64 mmol) and pivalate (2-chloro-4- (3-nitrophenoxy) ) -7H-pyrrolo [2,3-d] pyrimidin-7-yl) methyl (1.55 g, 3.83 mmol), potassium carbonate (1.35 g, 9.77 mmol), tris (dibenzylideneacetone) dipalladium (173 mg, 0.19 mmol) and dicyclohexyl ( 2 ', 4', 6'-triisopropylbiphenyl-2-yl) phosphine (222 mg, 0.47mmol), magnetite, and t-BuOH (35 mL) was heated to boiling and stirred under nitrogen for 2 h. The mixture was cooled to 40 ° C. 50 ° C and filtered through diatomaceous earth, washing with ethyl acetate (50 mL). The filtrate was concentrated under reduced pressure. the crude material was purified by column chromatography (DCM / MeOH = 50/1 as eluent) to give the title compound (1.7 g, 74% yield, M + H⁺= 608.3) as a light yellow solid.

SynthesisN1- (4- (3-aminophenoxy) -7H-pyrrolo [2,3-d] pyrimidin-2-yl) -N4- (1- (fluoroethyl) azetidin-3-yl) -2-methoxybenzene-1,4 -diamines [0314]

<img file="PL2880035T3_D0076.tif" />

A mixture of pivalate (2- (4- (1- (2-fluoroethyl) azetidin-3-ylamino) -2-methoxyphenylamino) -4- (3-nitrophenoxy) -7H-pyrrolo [2,3-d] pyrimidin-7. -yl) methyl (530 mg, 0.87 mmol), NHNH (98%, 2.5 mL), Pd / C (110 mg), magnetite, and MeOH (10 mL) was stirred at reflux overnight. The mixture was cooled to room temperature, and filtered through diatomaceous earth, washing with MeOH (20 mL). The filtrate was concentrated under reduced pressure. NaHCO 3 (aq) was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic layers were concentrated under reduced pressure. the crude material was purified by column chromatography (DCM / MeOH = 40/1 as eluant) to give the title compound (125 mg, 31% yield, M + H⁺= 464.2) as a white solid.

Synthesis of N- (3- (2- (4- (1- (2-fluoro-ethyl) -azetidin-3-ylamino) -2-methoxyphenylamino) -7H-pyrrolo [2,3-d] pyrimidin-4-yloxy) phenyl) acrylamide ( Compound 34)

<img file="PL2880035T3_D0077.tif" />

A 50mL round bottom flask with magnetite was charged with N1- (4- (3-aminophenoxy) -7H-pyrrolo [2,3-d] pyrimidin-2-yl) -N4- (1- (2-fluoro-ethyl) -azetidin-3-yl ) -2-methoxybenzene-1,4-diamine (125mg, 0.27 mmol), diisopropylethylamine (43 mg, 0.33 mmol) and DCM (20 mL). The mixture was cooled in an ice bath until the temperature was lower than 0 ° C, and a solution of acryloyl chloride (33 mg, 0.33 mmol) in THF (2 mL) was added dropwise over 5 minutes. The title compound is isolated and purified by preparative HPLC or preparative LC / MS, or other standard purification techniques. In some experiments, the title compound was isolated and purified by preparative LC / MS.

[0318] Compound 34 was also synthesized using a different synthetic route as follows:

<img file="PL2880035T3_D0078.tif" />

Synthesis of 34-B:

[0319] To compound 34-A (0.6g,) in a 100 mL round bottom flask was added Fe powder (0.3 g) and NH 4 Cl (0.5 g) in EtOH (60 mL). The reaction mixture was stirred at 90-100 for 3 ~ 4 h. At this point, the reaction was completed as indicated by TLC (DCM: MeOH = 8: 1). The reaction mixture was filtered through celite, and washed further with MeOH (~ 60 mL). The combined filtrate was concentrated under reduced pressure. The residue (oil) was dissolved in ethyl acetate (100 mL), washed with brine (50 mL x 2) and dried over sodium sulfate. The organic layer was concentrated under reduced pressure. The obtained crude product was then purified by column chromatography (DCM: MeOH = 30: 1) to give the desired product 34-B (420 mg, M + H).⁺= 578.5).

Synthesis of 34-C:

[0320] To compound 34-B (288 mg) in a 100 mL round bottom flask, acrylic acid (41 mg) and EDCI (176 mg) were added in DCM (30 mL). The reaction mixture was stirred at 0 ° C (ice bath) for 1 ~ 1.5 h. At this point, TLC (DCM: MeOH = 7: 1) showed the reaction was complete. A small amount of water (0.5 mL) was added to quench the reaction. The reaction mixture was concentrated under reduced pressure. The obtained residue was dissolved in ethyl acetate (30 mL). The organic layer was washed with brine (10 mL x2), dried over sodium sulfate and concentrated under reduced pressure to give the crude product which was further purified by column chromatography (DCM: MeOH = 30: 1) to give the desired product 34-C (79 mg). , M + H⁺= 632.5).

Synthesis of compound 34:

[0321] To compound 34-C (79 mg) in MeOH (15 mL) in a 100 mL round bottom flask, aqueous NaOH (2.5 mol / L) was added. The reaction mixture was stirred at 0 ° C (ice bath) for 4 ~ 5 h. At this point, LC-MS showed complete completion of the reaction. The reaction mixture was poured into water (100 mL), extracted with ethyl acetate (50 mL x3). The organic layer was washed with brine (30 mL x 2), dried over sodium sulfate and concentrated in vacuo. The crude product was further purified by column chromatography (DCM: MeOH = 25: 1) to give the desired product 34 (32 mg, M + H).⁺= 464.5).

[0322] Additional exemplary compounds not exemplified in these synthetic examples are prepared from the appropriate starting materials using methods analogous to those described in the preceding examples and schemes.

Biological Example A:

Cell-based in vitro screening using real-time electronic cell detection (RT-CES) [0323] Some assays and examples demonstrating the anti-cancer effects of compounds of the embodiments are described below.

[0324] Pyrrolopyrimidine compounds in the examples are developed for antitumor activity against tumor cells with specific molecular targets, i.e. EGFR (epidermal growth factor receptor). The antitumor efficacy of pyrrolopyrimidine compounds can be pre-screened in vitro using an EGFR tumor cell panel using a real-time electronic cell detection system from ACEA Biosciences, Inc. (or the xCELLigence system from Roche Applied Sciences / ACEA Biosciences Inc.), which provides dynamic information about the response of cells when they are exposed to an anticancer agent.

[0325] Details of this electronic cell detection technology, called electronic real-time detection (RT-CES^®) and related devices, systems and methods of their use are described in US Patent No. 7,732,127; patent number 7,192,752; patent number 7,459,303; patent number 7,468,255; patent number 7,470,533; U.S. Patent No. 7,560,269; interim patent application No. 60 / 397,749, filed July 20, 2002; US Patent Application Publication No. 60 / 435,400, filed December 20, 2002; the provisional application No. 60 / 469,572, filed May 9, 2003, PCT application number PCT / US03 / 22557 filed on July 18, 2003; PCT application number PCT / US03 / 22537 filed on July 18, 2003; PCT application number PCT / US04 / 37696, filed on November 12, 2004; PCT application number PCT / US05 / 04481 filed on February 9, 2005; U.S. Patent Application No. 10 / 705,447, filed November 10, 2003; U.S. Patent Application No. 10 / 705,615, filed November 10, 2003; U.S. Patent Application No. 10 / 987,732, filed November 12, 2004; U.S. Patent Application No. 11 / 055,639, filed on February 9, 2005, each of which is incorporated by reference.

Additional details on RT-CES technology are further disclosed in US provisional application number 60 / 519,567, filed November 12, 2003, and US Patent Application Publication No. 60 / 542,927, filed February 9, 2004, US Provisional Application No. 60 / 548,713, filed February 27, 2004, US Provisional Application No. 60 / 598,608, filed August 4, 2004; interim patent application No. 60 / 598,609, filed on August 4, 2004; US Patent Application Publication No. 60 / 613,749, filed on September 27, 2004; US Patent Application Publication No. 60 / 613,872, filed September 27, 2004; interim patent application No. 60 / 614,601, filed on September 29, 2004; US Patent Application Publication No. 60 / 630,071, filed November 22, 2004; provisional US patent application number 60/630, 131, filed November 22, 2004. [0326] For cell-substrate or cell-electrode impedance measurement using RT-CES technology, microelectrodes having appropriate geometries are performed on the lower surface portions of the microtiter plate or similar device, directed to the wells. The cells are inserted into the wells of the devices and cause contact and attach to the surface of the electrodes. The presence, absence or change of cell properties affects the electrode and ionic transition on the surfaces of the electrode sensor. Measurement of the impedance between two or more electrodes provides information on the biological state of the cells present on the sensors. When there are changes in the biological state of the cell analogue,

[0327] In the RT-CES system, the cell index is automatically derived and reported based on measured electrode impedance values. The cellular index obtained for a given well reflects: 1) how many cells are attached to the surface of the electrodes in this well; and 2) how well the cells are attached to the surface of the electrodes in this well. Thus, the more cells of the same type under similar physiological conditions adheres to the surface of the electrodes, the larger the cellular index. And the better the cells adhere to the surface of the electrodes (e.g., the cells decompose more to have larger contact areas, or the cells adhere more tightly to the surface of the electrodes), the larger the cellular index. We discovered

[0328] By using the RT-CES system, the pyrrolopyrimidine compounds described in the above examples have been shown to induce a similar impedance profile of cell responses in the RT-CES system to that generated by the positive control inhibitors. In addition, these compounds have been shown to inhibit EGFR-induced (epidermal growth factor receptor) cell migration in several cell lines. In addition, these compounds showed no or negligible effects when they were used in the treatment of cMet-independent cancer cell lines.

[0329] The RT-CES system (or RTCA xCELLigence system) comprises three elements, an electronic sensor analyzer, a device station and devices with a 16X microtiter plate, or

96X (i.e. E-Plate 16 or E-Plate 96). The microelectrode sensor array was created on slides with microlithographic methods and slides containing the electrode were mounted on plastic trays to form wells containing the electrode. Each device with a 16X (or 96X) microtiter plate used in the RT-CES system contained up to 16 (or 96) such wells containing an electrode. The device station receives devices with microtiter plates 16X or 96X and can switch electronically any one of the wells to the sensor analyzer for impedance measurement. During operation, devices with cells grown in wells are placed in the device station (SP RCTA xCELLigence station or SP RT-CES station), which is located inside the incubator. The electrical wiring connects the device station to the sensor analyzer (RTCA xCELLigence analyzer or RTCES analyzer). Under the control of the RT-CES software or RTCA xCELLigence, the sensor analyzer can automatically select the measuring wells and carry out impedance measurements on a continuous basis. The impedance data from the analyzer are transferred to the computer, analyzed and processed by the integrated software.

[0330] The impedance measured between the electrodes in a single well depends on the geometry of the electrodes, the concentration of ions in the well and whether the cells are attached to the electrode. In the absence of cells, the electrode impedance is determined mainly by the ionic environment at both the electrode / solution interface and the mass of the solution. In the presence of cells, the cells attached to the electrode sensor surface will change the local ion environment at the electrode / solution interface leading to an increase in impedance. The more cells on the electrodes, the greater the increase in electrode-cell impedance. In addition, the change in impedance also depends on cell morphology and the extent to which cells attach to the electrodes.

[0331] In order to quantify the condition of cells by measuring the cell-electrode impedance, a parameter is designated as a Cell Index according to

CI = maxf |

J, where Rb (f) and Rcell (f) are frequency dependent electrode resistances (impedance component) respectively without cells or with existing cells. N is the number of frequency points at which the impedance is measured. Thus, the Cell Index is a quantitative measure of the condition of cells in a well containing an electrode. Under the same physiological conditions, more cells attached to the electrodes lead to a higher Rcell (f) value, which leads to a higher value for the Cell Index. In addition, for the same number of cells present in the well, a change in cell status such as morphology will lead to changes in the Cell Index. For example, an increase in cell adhesion or the spread of cells leads to a larger cell-electrode contact surface, which will lead to an increase in Rcell (f) and thus a higher value of the Cell Index. The Cell Index can also be calculated using a pattern that is different from that described herein. Other methods of calculating the Cell Index based on electrode impedance measurement can be found in US Patent No. 7,732,127; patent number 7,192,752; patent number 7,459,303; patent number 7,468,255; patent number 7,470,533; U.S. Patent No. 7,560,269; PCT application number

PCT / US04 / 37696, filed November 12, 2004, PCT Application No. PCT / US05 / 04481, filed February 9, 2005, US Patent Application No. 10 / 987,732, filed November 12, 2004, US Patent Application No. 11 / 055,639, filed February 9, 2005 .

Biological Example B-1

Biological activity of pyrrolopyrimidine compounds on cell lines with mutant EGFR

Material and methods

Cell culture and reagents [0332] All cell lines were obtained from the American Tvpe Culture Collection and were incubated at 37 ° C with 5% CO 2 in medium supplemented with 10% fetal bovine serum and 1% L-glutamine-penicillin-streptomycin. H1975 and HCC827 cells were cultured with RPMI 1640 media. A431 cells were maintained in Dulbecco's Modification of Eagle's Medium. EGF (R &amp; D), EGF inhibitor was resuspended and stored in accordance with the manufacturer's instructions.

Cell proliferation assay and growth inhibition [0333] Cell proliferation was assessed using the WST assay (Roche, Indianapolis, IN) according to the manufacturer's instructions. H1975, HCC827 and A431 cells were loaded with 3,000, 3,000 and 4,000 cells per well in 96-well plates, and after 24-hour incubation, cells were treated with test compounds for 72 hours. Cell viability was determined by incubating cells with a WST-1 detector for 2 hours, then with absorbance measurement at 450nm. Data were calculated using GraphPad Prism version 4.0. IC 50 values ​​were fitted using a non-linear regression model with a sigmoidal dose response.

Western blotting [0334] H1975 and A431 cells were seeded in 6-well plates at a concentration of 1 ^χ 10⁶ cells per well. After 24 hours growth in serum-containing medium, the cells were incubated in serum-free media for 1 hour and then treated with the test compound for 2 hours. A431 cells were stimulated with 30ng / mL EGF over the last 20 minutes of compound treatment. Western blots were performed on whole cell extracts using EGFR phospho-specific antibodies (pY1068), total EGFR, phospho-Akt (Ser-473), total Akt, phospho-ERK1 / 2 (pT202 / pY204) and total ERK1 / 2 (Cell Signaling Technologv ).

[0335] Tumor sections were snap frozen in liquid nitrogen for protein isolation, and EGFR signal transduction was assessed by Western blot with primary antibodies including: phospho-specific EGFR (pY1068), total EGFR, phospho-Akt (Ser-473), total Akt, phospho-ERK1 / 2 (pT202 / pY204) and total ERK1 / 2.

ELISA test [0336] H1975 and A431 cells were plated on each well of a 96-well plate at a density of 4^χ10⁴cells per well. After 24 hours growth in serum-containing medium, the cells were treated with the test compound in serum-free medium for 2 hours. A431 cells were stimulated with 30ng / mL EGF over the last 15 minutes of compound treatment. The cells were washed with ice-cold PBS and then extracted with 100 μΐ per well of cell lysis buffer. EGFR phosphorylation was measured using an ELISA sandwich ELISA with EGFR specific antibody pair (pY1068) and total EGFR.

Results

Compound 3 inhibits the proliferation of EGFR mutant cells [0337] The following compounds were tested.

<img file="PL2880035T3_D0079.tif" />

<img file="PL2880035T3_D0080.tif" />

<img file="PL2880035T3_D0081.tif" />

<img file="PL2880035T3_D0082.tif" />

[0338] Sensitivity of tumor cell lines that express EGFR WT, Exon 19 Del, L858R / T790M and delE746-A750 to Compound 3, Compound B and gefitinib. Cell proliferation assays were performed with increasing concentrations of compounds for 72 hours using WST. IC 50 values ​​were determined using GraphPad software. Compound 3 inhibits the proliferation of T790M-positive H1975 cells more strongly than gefitinib.

Table 1

<td>Relationship</td><td>Mobile H1975 (T790M / L858R)</td><td>Cell A431 (WT)</td><td>Cell HCC827 (delE746-A750)</td>

<td>Compound 3</td><td>0.61 μΜ</td><td>10.8 μΜ</td><td>0.019 μΜ</td>

<td>Compound B</td><td>1.1 μΜ</td><td>4.5 μΜ</td><td>0.013 μΜ</td>

<td>gefitinib</td><td>&gt; 10 μΜ</td><td>ND</td><td>0.024 μΜ</td>

Compound 3 inhibits EGFR phosphorylation in H1975 cells [0339] Inhibition of EGFR phosphorylation and proliferation in H1975 cells treated with Compound 3. H1975 and A431 cells were incubated with various concentrations of Compound 3 or Compound B for 2 hours, and whole cellular extracts were directly harvested and tested for pEGFR according to ELISA. IC 50 values ​​were determined using GraphPad software.

Table 2

<td>Relationship</td><td>Mobile H1975 (T790M / L858R)</td><td>Mobile A431 (WT)</td>

<td>Compound 3</td><td>0.031 μΜ</td><td>12.7 μΜ</td>

<td>Compound B</td><td>0.063 μΜ</td><td>8.9 μΜ</td>

Compound 3 inhibits the EGFR signaling pathway in H1975 cells [0340] Exponentially growing lung cancer cell types H1975 were treated with Compound 3 at the indicated concentrations for 2 hours in serum free media. As shown in Figure 1, whole cell extracts were separated by SDS-PAGE before transfer to nitrocellular membranes. Inhibition of phosphorylation leads to EGFR inhibition of its downstream effectors p-Akt and p-ERK. All antibodies were obtained from Cell Signaling.

Compound 3 inhibits the EGFR signaling pathway in H1975 tumors [0341] Compound 3 was given PO at 100mg / kg, and tumors were sampled at 1, 4, 8, 18 and 25 hours after a single dose. As shown in Figure 2, immunoblots were probed for pEGFR, total EGFR, pAct, total Akt, p-ERK and total ERK. Compound 3 inhibited EGFR phosphorylation in a time-dependent manner and inhibition at EGFR leads to inhibition of its downstream effectors p-Akt and p-ERK.

Comparison of Compound 3 and Compound A

1. The result of JTS [0342]

<td colspan="2">this</td><td colspan="2">bale 3</td>

<td>Relationship</td><td>Mobile H1975 (T790M / L858R)</td><td>Mobile A431 (WT)</td><td>Cell HCC827 (delE746-A750)</td>

<td>Compound 3</td><td>0.73 μΜ</td><td>0.62 μΜ</td><td>0.011 μΜ</td>

<td>Relationship</td><td>Mobile H1975 (T790M / L858R)</td><td>Mobile A431 (WT)</td><td>Cell HCC827 (delE746-A750)</td>

<td>Compounds A</td><td>1.63 μΜ</td><td>4.17 μΜ</td><td>0.023 μΜ</td>

2. ELISA result [0343]

Table 4

<td>Relationship</td><td>Mobile H1975 (T790M / L858R)</td><td>Cell A431 (WT)</td><td>Cell H1975 (T790M / L858R) EGF stimulation</td>

<td>Compound 3</td><td>0.0032 μΜ</td><td>0.4737 μΜ</td><td>0.025 μΜ</td>

<td>Compound A</td><td>0.0088 μΜ</td><td>1.0270 μΜ</td><td>0.091 μΜ</td>

Biological Example B-2 Cell Culture and Reagents [0344] All cell lines were obtained from the American Type Culture Collection and maintained at 37 ° C with 5% CO 2, in a medium supplemented with 10% fetal bovine serum and 1% L-glutamine-penicillin-streptomycin. . H1975 and HCC827 cells were cultured with RPMI 1640 media. A431 cells were maintained in Dulbecco's Modification of Eagle's Medium. GTL-16 cells, T47D cells and BxPC3 cells were cultured with RPMI 1640 media. NIH-3T3 cells, H460 cells and HepG2 cells were cultured with Dulbecco's Modification of Eagle's Medium. A549 cells were cultured with F-12K Nutrient Mixture media. H295R were cultured with DMEM medium: F12 Media. WST-1 reagent was obtained from Roche. EGF, EGF (R &amp; D) inhibitors were resuspended and stored according to the manufacturer's instructions.

Cell proliferation assay and growth inhibition [0345] Cell proliferation was evaluated using the WST assay (Roche, Indianapolis, IN) according to the manufacturer's instructions. H1975, HCC-827 and A431 cells were seeded at 3,000, 3,000, and 4,000 cells per well in 96-well plates, and after 24h, cells were treated with test compounds for 72 hours. NIH-3T3 cells, A549 cells, H295R cells, GTL-16 cells, H460 cells, HepG2 cells, Hela cells, T47D cells and BxPC3 cells were plated at 2,000, 2,000, 5,000, 5,000, 2,500, 5,000, 2,000, 5,000 and 5,000 cells per well in 96-well plates. Cell viability was determined by incubating cells with WST-1 reagent for 3 hours. The absorbance was measured at OD450-620 using a Beckman DTX880. Data were calculated using GraphPad Prism version 4.0.

ELISA assays [0346] H1975, HCC-827 and A431 cells were seeded in a 96-well plate at a density of 40,000, 40,000 and 60,000 cells per well, respectively. After 24 hr growth in serum-containing medium, the cells were treated with the test compound in serum-free medium for 2 h. A431 cells were stimulated with 50ng / mL EGF during the last 15min treatment with the compound. The cells were washed with ice-cold PBS and then extracted with 100 μΐ per well of cell lysis buffer. EGFR phosphorylation was measured using an ELISA sandwich ELISA with EGFR specific antibody pair (pY1068) and total EGFR. Data were calculated using GraphPad Prism version 4.0. The IC50 was fitted using a non-linear regression model with a sigmoidal dose response.

Western Blotting [0347] H1975, HCC-827 and A431 cells were seeded in 6-well plates at a concentration of 1 ^x10⁶ cells per well. After 24 hr growth in serum-containing medium, the cells were incubated in serum-free media for 1 h and then treated with the test compound for 2 h. A431 cells were stimulated with 30ng / mL EGF over the last 20 min compound treatment. Western blots were performed on whole cell extracts using phosphor-specific EGFR (pY1068) antibodies, total EGFR, phospho-Akt (Ser-473), total Akt, phospho-ERK1 / 2 (pT202 / pY204) and total ERK1 / 2 (Cell Signaling Technology ). The density of the blotting bands was obtained using Image J software, and the IC50 Tyr1068 EGFR phosphorylation was adjusted using a non-linear regression model using GraphPad Prism version 4.0.

[0348] Compound 3 was administered orally at the indicated dose (12.5, 50, 200mg / kg), and Gefitinib (GF) was administered orally at 100mg / kg. Tumor tissues were collected 1, 4, 8, and 24 hours after Day 1 and after a single dose, or were collected on Day 8 and after 8 sub-doses for the dose (12.5, 50 mg / kg). Tumor sections were snap frozen in liquid nitrogen for protein isolation, and EGFR signal transduction was assessed by Western blot with primary antibodies including the following: phospho-specific EGFR (pY1068), total EGFR.

Cell-based "pulse-chase" assay to evaluate the irreversibility of compound [0349] H1975 was seeded at 3,000 cells per well in an RTCA system (xCELLigence SP instrument, ACEA Biosciences). After one day of culture, the cells were treated with Compound 3 compound WZ4002 at a concentration of 10μΜ for 22 hours and then removed compared to overtaxed drugs. About 60 hours after regeneration, the cell is subject to WST lifetime measurement.

Results

Compound 3 inhibited the proliferation of tumor cells harboring the EGFR mutation.

[0350] Compound 3 achieved inhibition of H1975 cell proliferation (T790M / L858R) with IC50 at 91 ± 60nM, and with IC50 at 19 ± 8 nM for HCC827 cells (Del E746-A750), while cell sensitivity A431 (WT) is significantly lower (IC 50 = 2113 ± 1660 nM). In contrast, gefitinib, a first-generation EGFR inhibitor, was sensitive to A431 cells but had no activity in inhibiting the proliferation of T790M-bearing cells (IC50&gt; 20uM).

<td>Relationship</td><td>Mobile H1975 (T790M / L858R)</td><td>Mobile A431 (WT)</td><td>Cell HCC827 (DelE746-A750)</td>

<td>Compound 3</td><td>91 ± 60 nM</td><td>2113 ± 1660 nM</td><td>19 ± 8 nM</td>

<td>WZ4002</td><td>1905 ± 732 nM</td><td>4393 ± 617 nM</td><td>35 ± 12 nM</td>

<td>gefitinib</td><td>&gt; 20,000 nM</td><td>523 ± 115 nM</td><td>9 ± 1 nM</td>

Compound 3 significantly reduced Tyr1068 EGFR phosphorylation in EGFR mutant cells [0351] H1975 and A431 cells were incubated with different concentrations of Compound 3 or WZ4002 for 2 h, and all cellular extracts were directly harvested and tested for pEGFR by ELISA. IC 50 values ​​were determined using GraphPad software.

[0352] A cell-based ELISA verified that compound 3 significantly reduced Tyr1068 EGFR phosphorylation in EGFR mutant cell lines, whereas

Gefitinib showed inhibition of phosphorylation to a much lesser degree.

<td>Relationship</td><td>Mobile H1975 (T790M / L858R)</td><td>Mobile A431 (WT)</td>

<td>compound 3</td><td>4 ± 2 nM</td><td>650 ± 63 nM</td>

<td>WZ4002</td><td>32 ± 12 nM</td><td>970 ± 340nm</td>

[0353] As shown in the following table, Compound 3 significantly reduced Tyr1068 EGFR phosphorylation and downstream signaling in cells with an EGFR mutant and is less effective in a cell line expressing wild-type EGFR.

<td rowspan="2">EGFR genotype</td><td rowspan="2">Cell line</td><td colspan="3">IC50 (nM) by WB (Tyr1068 phospho)</td>

<td>compound 3</td><td>gefitinib</td><td>WZ4002</td>

<td>T790M / L858R</td><td>H1975</td><td>4.4</td><td>860</td><td>21</td>

<td>DelE746-A750</td><td>HCC-827</td><td>9.8</td><td>5.4</td><td>58</td>

<td>Wild type</td><td>A431</td><td>288</td><td>1.6</td><td>53</td>

<td colspan="2">Czułość (A431 / H1975)</td><td>65X</td><td>0.002Χ</td><td>2.5X</td>

[0354] As shown in Figures 9A and 9B, Compound 3 inhibited phosphorylation of EGFR-Tyr1068 and downstream signaling in cells with mutant EGFR H1975. Comparative data for Gefitinib and WZ4002 are shown in Figures 9C-9F.

[0355] As shown in Figures 10A and 10B, Compound 3 inhibited phosphorylation of EGFR-Tyr1068 and downstream signaling in cells with mutant EGFR HCC-827. Comparative data for Gefitinib and WZ4002 are shown in Figures 10C-10F.

Compound 3 inhibited EGFR phosphorylation in H1975 tumors.

[0356] As shown in Figures 11A and 11B, Compound 3 was less effective in inhibiting the phosphorylation of EGFR-Tyr1068 and further signaling in A431 expressing cells.

WT EGFR. Comparative data for Gefitinib and WZ4002 are shown in Figures 11C-11F.

[0357] Compound 3 significantly inhibited EGFR phosphorylation in tumor tissues of H1975, in all three doses of 12.5, 50 and 200 mg / kg. The inhibition of EGFR phosphorylation by Compound 3 was dose dependent and time dependent. In contrast, inhibition of EGFR phosphorylation was not detected for gefitinib at a dose of 100mg / kg.

[0358] As shown in Figure 12, Compound 3 inhibits EGFR phosphorylation in tumor tissues of H1975 at a single dose of compound 3.

[0359] As shown in Figure 13, Compound 3 inhibits EGFR phosphorylation in tumor tissues of H1975 after 8 consecutive doses of compound 3.

[0360] As shown in Figure 14, Compound 3 irreversibly inhibited the proliferation of H1975 cells harboring the T790M EGFR mutation. Reversibility of Compound 3 was assessed in a cell-based, pulse-chase test. As shown in Figure 14, after withdrawal of compound 3 after 22-hour treatment, inhibition of H1975 proliferation persisted (7.8 ± 1.3%) to 60 hours. However, regeneration after WZ4002 treatment was 27 ± 10%. The results of this study showed that Compound 3 is an irreversible EGFR inhibitor and has strong binding properties than WZ4002.

Service life compared to substrate control (%) [03611___

<td></td><td>compound 3 for the treatment of 22 hours</td><td>WZ4002 for the treatment of 22 hours</td>

<td>Cell viability H1975 (%)</td><td>7.8 ± 1.3</td><td>27 ± 10</td>

1. The result of JTS [03621

<td>Relationship</td><td>Mobile H1975 (T790M / L858R)</td><td>Cell A431 (WT)</td><td>Cell HCC827 (delE746-A750)</td>

<td>compound 3</td><td>0.19 μM</td><td>2.03 μM</td><td>0.011 μM</td>

<td>compound A</td><td>1.16 μM</td><td>11.82 uM *</td><td>0.023 μM</td>

<td colspan="4">* Note: The A431 value for Compound A was previously reported incorrectly as 9.14 μM.</td>

2. ELISA result [03631

<td>Relationship</td><td>Mobile H1975 (T790M / L858R)</td><td>A431 cell phone (WT) EGF stimulation</td><td>Mobile H1975 (T790M / L858R)</td>

<td>compound 3</td><td>0.0032 μM</td><td>0.4737 μM</td><td>0.025 μM</td>

<td>compound A</td><td>0.0088 μM</td><td>1.0270 μM</td><td>0.091 μM</td>

Biological Example B-3 [0364] Using the new H1975 cell line and the ELISA protocol described above in Biological Example B-2) the following results were obtained:

ELISA test

<td>IC50</td><td>(UM)</td><td>IC50 (uM)</td><td>IC50</td><td>Selektywnoś</td><td colspan="2">Selektywnoś</td>

<td>H1975</td><td></td><td>Hcc827</td><td>(UM)</td><td>ć A431 /</td><td colspan="2">ć A431 /</td>

<td colspan="2">T790M / L858R</td><td>(DelE746-A750)</td><td>A431WT</td><td>H1975</td><td>Hcc827</td><td></td>

<td>(lack</td><td></td><td>(no stmulation</td><td>(stvmulac</td><td></td><td></td><td></td>

<td colspan="2">stvmulacji</td><td>EGF)</td><td>I EGF)</td><td></td><td></td><td></td>

<td>EGF)</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>0.0018</td><td></td><td>.0076</td><td>.1233</td><td>68.5000</td><td>16.2237</td><td></td>

<td>Zwk 3 0.0016</td><td></td><td>.0072</td><td>.1300</td><td>81.2500</td><td>18.0556</td><td></td>

<td>0.0010</td><td></td><td>.0072</td><td>.0901</td><td>90.1000</td><td>12.5139</td><td></td>

<td colspan="2">Dia. ± SD</td><td>0.0015 0.0073</td><td>0.1145 ±</td><td>79.9500 ±</td><td>15.5977</td><td>±</td>

<td></td><td></td><td>± ±</td><td>.0174</td><td>10.8585</td><td>2.8233</td><td></td>

<td></td><td></td><td>0.0004 0.0002</td><td></td><td></td><td></td><td></td>

<td>Zwk B</td><td></td><td>0.0299 0.0419</td><td>.6184</td><td>20.6823</td><td>14.7589</td><td></td>

<td>(WZ4002)</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td></td><td>0.0257 0.0385</td><td>.6592</td><td>25.6498</td><td>17.1221</td><td></td>

<td></td><td></td><td>0.0212 0.0391</td><td>.6097</td><td>28.7594</td><td>15.5934</td><td></td>

<td>Dia. ±</td><td>.0256</td><td>± 0.0399 ±</td><td>.6291</td><td>± 25.0305</td><td>15.8248</td><td></td>

<td>SD</td><td>0.0043</td><td>0.0018</td><td>.0264</td><td>± 4.0740</td><td>± 1.1984</td><td></td>

<td>P values ​​(Zwk 3</td><td>0.0007</td><td>0.000007</td><td>0.000013</td><td>0.0012</td><td>.9042</td><td></td>

<td>and Zwk B)</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Zwk A</td><td>.0145</td><td>.0411</td><td>.5641</td><td>38.9034</td><td>13.7251</td><td></td>

<td></td><td>.0159</td><td>.0395</td><td>.4794</td><td>30.1509</td><td>12.1367</td><td></td>

<td></td><td>.0040</td><td>.0305</td><td>.2884</td><td>72.1000</td><td>9.4557</td><td></td>

<td>Dia. ±</td><td>.0115</td><td>± 0.00370 ±</td><td>.4440</td><td>± 47.0515 ±</td><td>11.7725</td><td>±</td>

<td>SD</td><td>.0065</td><td>.0057</td><td>.1412</td><td>22.1297</td><td>2.1578</td><td></td>

<td>P * (Zwk</td><td>.0565</td><td>0.00085</td><td>0.01618</td><td>.0819</td><td>0.13569</td><td></td>

<td>3 and Zwk A)</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2">[0365] Using the new lines</td><td>cellular H1975</td><td colspan="2">and the WST protocol described</td><td>above</td><td>(in</td>

<td colspan="4">Biological Examples B-2 with the presence of 5% fetal</td><td colspan="3">bovine serum), was obtained</td>

<td>the following results:</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="3">WST test (cell viability test)</td><td></td><td></td><td></td><td></td>

<td></td><td>IC50 H1975_ T790M</td><td>(UM) 5% FBS / L858R</td><td>IC50 (UM) Hcc827 (DelE746A750)</td><td>IC50 (uM) A431 WT</td><td>selectivity A431 / H1975</td><td>selectivity A431 / Hcc827</td>

<td></td><td>0.019</td><td></td><td>0.003</td><td>0692</td><td>36.42</td><td>230.67</td>

<td></td><td>0.018</td><td></td><td>0.008</td><td>0526</td><td>29.22</td><td>65.75</td>

<td>ZWK</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>3</td><td>0.019</td><td></td><td>0.005</td><td>0694</td><td>36.53</td><td>138.80</td>

<td>Average</td><td colspan="2">0.019 ± 0.001</td><td>0.005 ±</td><td>0637 ±</td><td>34.057 ±</td><td>145.072 ±</td>

<td>± SD</td><td></td><td></td><td>0.002</td><td>0.096</td><td>4.187</td><td>82637</td>

<td>ZWK</td><td>0.053</td><td></td><td>0.007</td><td>0901</td><td>17.00</td><td>128.71</td>

<td>(W-Z CAKE</td><td>0.080</td><td></td><td>0.021</td><td>1027</td><td>12.84</td><td>48.90</td>

<td>4002)</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>0.106</td><td></td><td>0.013</td><td>1.025</td><td>9.67</td><td>78.85</td>

<td>Average</td><td colspan="2">0.080 ± 0.026</td><td>0.014 ±</td><td>0984 ±</td><td>13.169 ±</td><td>85,488 ±</td>

<td>± SD</td><td></td><td></td><td>0.007</td><td>0.072</td><td>3.676</td><td>40317</td>

<td>P values</td><td>.0163</td><td></td><td>.1251</td><td>0.0075</td><td>0.0029</td><td>.3245</td>

<td>(Zwk 3 and Zwk</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>B)</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>0.106</td><td></td><td>0.028</td><td>2,914</td><td>27491</td><td>104071</td>

<td></td><td>0.111</td><td></td><td>0.059</td><td>2.617</td><td>23577</td><td>44356</td>

<td>ZWK</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>AND</td><td>0.067</td><td></td><td>0.035</td><td>2809</td><td>41925</td><td>80257</td>

<td>Average</td><td colspan="2">0.095 ± 0.024</td><td>0.041 ±</td><td>2.780 ±</td><td>30.998 ±</td><td>76.228 ±</td>

<td>± SD</td><td></td><td></td><td>0.016</td><td>0.151</td><td>9664</td><td>30,061</td>

<td>P * values</td><td>0.0055</td><td></td><td>.0205</td><td>3.18539E-</td><td>.6414</td><td>.2466</td>

<td>(Zwk 3 and Zwk</td><td></td><td></td><td></td><td>05</td><td></td><td></td>

<td>AND)</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="7">* p-values ​​1) &lt;0.05 (marked in black bold) means a significant difference;</td>

<td colspan="4">2) 0.05-0.15 means an important difference / improvement</td><td colspan="2">but not significant; 3)&gt; 0.15</td><td>means none</td>

<td>significant difference</td><td></td><td></td><td></td><td></td><td></td><td></td>

[0366] Based on the experimental data and p-values ​​in the two above tables, Compound 3 showed significantly greater potency than Compounds A and B in both WST and ELISA with ten (10) p values ​​&lt;0.05 and two (2) p values ​​between 0.05 - 0.15. In terms of selectivity, Compound 3 showed a general trend of higher selectivity than Compounds A and B in both WST and ELISA with two (2) p values ​​&lt;0.05, two (2) p values ​​between 0.05-0.15 and four (4) p values ​​over 0.15.

Biological Example C

Evaluation of the efficacy of Compound 3 in Treatment H1975, HCC827 and A431

Mice Heteropterous Models [0367] This example evaluates the efficacy of Compound 3 in the treatment of human non-small lung NPC-H1975 (L858R / T790M) non-small cell lung tumor models, human HCC827 lung adenocarcinoma (L858R) and human A4 epidermoid skin cancer (WT) in nude mice. . Gefitinib, the first generation of a reversible EGFR tyrosine kinase inhibitor, was used as a positive control in these three mouse tumor xenograft models.

Experimental Design and Dosage Scheme [0368] Experiment design and dosage scheme are outlined below.

<td colspan="5">Table 5: M</td><td>Model NCI-H</td><td colspan="3">1975</td>

<td>Dec pa</td><td>n</td><td>treatment no</td><td>dose and (Mg / k g)</td><td>Volume dosage nia (μΐ / g)</td><td>Road dosage present</td><td>Solvents Score</td><td>Days for dosage present</td><td>schem at</td>

<td>1</td><td>8</td><td>basis e</td><td></td><td>16.7</td><td>AFTER</td><td>PEG system</td><td>14</td><td>QD</td>

<td>2</td><td>8</td><td>Relationship 3</td><td>25</td><td>10</td><td>AFTER</td><td>PEG system</td><td>14</td><td>QD</td>

<td>3</td><td>8</td><td>Relationship 3</td><td>50</td><td>10</td><td>AFTER</td><td>PEG system</td><td>14</td><td>QD</td>

<td>4</td><td>8</td><td>Relationship 3</td><td>100</td><td>16.7</td><td>AFTER</td><td>PEG system</td><td>14</td><td>QD</td>

<td>5</td><td>8</td><td>Gefitin b</td><td>100</td><td>10</td><td>AFTER</td><td>1% tween80</td><td>14</td><td>QD</td>

Table 6: Model HCC827

<td>Dec pa</td><td>n</td><td>treatment no</td><td>dose and (Mg / k g)</td><td>Volume dosage nia (μΐ / g)</td><td>Road dosage present</td><td>Solvents Score</td><td>Days for dosage present</td><td>schem at</td>

<td>1</td><td>8</td><td>basis e</td><td></td><td>10</td><td>AFTER</td><td>-</td><td>35</td><td>QD</td>

<td>2</td><td>8</td><td>Relationship 3</td><td>50</td><td>10</td><td>AFTER</td><td>PEG system</td><td>35</td><td>QD</td>

<td>3</td><td>8</td><td>tied k 3</td><td>50</td><td>10</td><td>AFTER</td><td>0.5% MC</td><td>35</td><td>QD</td>

<td>4</td><td>8</td><td>Gefitin b</td><td>100</td><td>10</td><td>AFTER</td><td>1% tween80</td><td>7</td><td>QD</td>

Table 7: Model A431

<td>Dec pa</td><td>n</td><td>treatment no</td><td>dose and (Mg / k g)</td><td>Volume dosage nia (μΙ / g)</td><td>Road dosage present</td><td>Solvents Score</td><td>Days for dosage present</td><td>schem at</td>

<td>1</td><td>8</td><td>basis e</td><td></td><td>16.7</td><td>AFTER</td><td>PEG system</td><td>14</td><td>QD</td>

<td>2</td><td>8</td><td>tied k 3</td><td>100</td><td>16.7</td><td>AFTER</td><td>PEG system</td><td>14</td><td>QD</td>

<td>3</td><td>8</td><td>Gefitin b</td><td>100</td><td>10</td><td>AFTER</td><td>1% tween80</td><td>14</td><td>QD</td>

<td colspan="9">Note: n: number of animals; Dosage volume: adjust the dosing volume based on body weight; PEG system: PEG200: alcohol: 5% dextrose = 4: 1: 5; The treatment regimen was adjusted if weight loss&gt; 15%.</td>

Keeping Animals

Animals [0369] Details of animals are shown below.

Genre: Mouse Strain: naga Nu / Nu Age: 7-8 weeks Gender: Female Body weight: 20-25 g

Animal Contributor: Vital River Laboratories, Beijing, China Linger Conditions [0370] Mice were housed in individual ventilated IVC (Individual Ventilation Cages) cages at constant temperature and humidity with 4 animals per each cage at ACEA Bioscience Hangzhou Inc.

- Temperature: around 20-26 ° C.

- Humidity about 40-70%.

[0371] IVC cages: Made of polycarbonate. The size is 300 mm x 180 mm x 150 mm. The substrate material is a corn cob that is changed twice a week.

[0372] Diet: The animals had free access to the irradiated sterilized food in the form of a dry granulate throughout the study period.

[0373] Water: Animals have free access to sterile drinking water.

[0374] Cage identification: Identification labels for each cage contained the following information: number of animals, sex, strain, date of receipt, treatment, study number, group number and start date of treatment.

[0375] Identification of animals: Animals were marked with ear incisions.

Experimental methods and procedures

Cell Culture [0376] NCI-H1975, HCC827 and A431 tumor cells were maintained in vitro as a monolayer culture in medium supplemented with 10% fetal bovine serum, 100U / ml penicillin and 100μg / ml streptomycin at 37 ° C in a 5% CO2 atmosphere. air in accordance with ATCC recommendations. Tumor cells were routinely passaged twice a week by trypsin-EDTA treatment. Cells growing in the exponential growth phase were collected and counted for tumor inoculation.

Tum Inoculation [0377] Each mouse was inoculated subcutaneously in the right flank with H1975 cells, respectively (5 x 10⁶), HCC827 (5 x 10⁶) and A431 (5 x 10⁶) in 0.2 ml of medium for tumor development. Treatment was started when the tumor size reached about 200-250 mm³. The test products were administered to mice according to a predetermined regimen, which is shown in the experimental design table.

Observations [0378] All animal, care and treatment procedures in this study were conducted in accordance with guidelines approved by the Institutional Animal Care and Use Committee (IACUC) according to the guidelines of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). During routine screening, the animals were checked for any effects of growth and treatment of the tumor on normal behavior such as mobility, food and water intake (by observation), increase / loss of body weight (body weight was measured twice a week), maturation of eyes /. hair and any other incorrect effect. Death and observed clinical symptoms were recorded based on the number of animals within each subset. Animals that have been observed to be in a constantly deteriorating condition,

Tumor Measurements and Endpoints [0379] The primary endpoint was to check whether tumor growth could be delayed or the mice could be cured. Tumor size was measured twice a week in two dimensions using a caliper and the volume was expressed in mm³ using the formula: V = 0.5 axb², where a and b are respectively long and short diameters. The size of the tumor was then used to calculate the values ​​of both TC and T / C. TC was calculated from T as the median time (in days) required for tumors from the treatment group to reach a predetermined size (e.g., 1,000 mm³), and C as the median of time (in days) for control group tumors to achieve the same size. T / C (in percent) was a measure of antitumor efficacy; T and C were the average volume of treatment and control groups, respectively, on a given day. The tumor mass was determined at the end of the study. The T / C value (in percent) was calculated, where T and C were mean tumor weights of treatment and control groups, respectively.

Statistical Analysis [0380] Summary statistics, including the mean and standard error of the mean (SEM), are given for the tumor volume in each group for each time point.

[0381] Statistical analysis of the difference in tumor volume and tumor mass between groups was based on data obtained at the best therapeutic time point after the last dose (15th day after tumor inoculation).

[0382] A one-way ANOVA was performed to compare tumor volume and tumor mass between groups and when a significant value of F statistic (treatment variance coefficient to error variance) was obtained, comparisons between groups were performed by the LSD and GamesHowell test. All data was analyzed using SPSS 16.0. p &lt;0.05 was considered statistically significant.

Results

5.1. Body Weights [0383] The results of body weight changes in tumor-bearing mice for models NCI-H1975, HCC827 and A431 are shown in Figures 3, Figure 4, and Figure 5, respectively.

[0384] The mouse's body weights in different groups of tumor-bearing mice at the end of treatment on NCI-H1975, HCC827 and A431 are shown in Table 8, Table 9 and Table 10 respectively.

Table 8. Mice Body Weights in Different Groups on the NCI-H1975 Model

<td>Treatment</td><td>Mouse Mass (g)^and on 23 (14)</td><td>p</td><td></td>

<td>Subsoil</td><td>22.99 ± 0.26</td><td></td><td></td>

<td>Compound 3 25 mg / kg after qd</td><td>22.28 ± 0.55</td><td>0.</td><td>364</td>

<td>Compound 3 50 mg / kg after qd</td><td>22.73 ± 0.33</td><td>0.</td><td>737</td>

<td>Compound 3 100 mg / kg after qd</td><td>22.53 ± 0.66</td><td>0.</td><td>555</td>

<td>Gefitinib 100 mg / kg after qd</td><td>20.43 ± 0.71</td><td>0.</td><td>002</td>

<td colspan="3">Table 9. Mice Body Weights in Different Groups on Model HCC827</td>

<td>Treatment</td><td>Mouse Mass (g)^and on 25 (14)</td><td>p</td>

<td>Subsoil</td><td>23.83 ± 0.71</td><td></td>

<td>Compound 3 (PEG) 50 mg / kg after qd</td><td>23.26 ± 0.47</td><td>0.523</td>

<td>Compound 3 (MC) 50 mg / kg after qd</td><td>23.54 ± 0.67</td><td>0.743</td>

<td>Gefitinib 100 mg / kg after qd</td><td>23.70 ± 0.43</td><td>0887</td>

Table 10. Mice Body Weights in Different Groups on A431 Model

<td>Treatment</td><td>Mouse Mass (g)^and on 25 (14)</td><td>p</td>

<td>Subsoil</td><td>25.64 ± 0.53</td><td></td>

<td>Compound 3 100 mg / kg after qd</td><td>25.66 ± 0.72</td><td>0979</td>

<td>Gefitinib 100 mg / kg after qd</td><td>21.56 ± 0.51</td><td>0.000</td>

Note: a. Medium ± SEM

Tumor volumes [0385] Tumor volumes of different groups at different time points on NCI-H1975, HCC827 and A431 are shown in Table 11, Table 12, and Table 13, respectively.

Table 11. Tumor sizes in the Different Treatment Groups on the NCI-H1975 model

Tumor volume (mm³)^and

<td rowspan="2">Days</td><td rowspan="2">Subsoil, QD</td><td rowspan="2">AFTER,</td><td colspan="3">Compound 3, Compound 3, Compound 3,</td><td rowspan="2">Gefitinib, PO, QD 100mpk</td>

<td>PO, QD 25mpk</td><td>PO, QD 50mpk</td><td>PO, QD 100mpk</td>

<td>9</td><td colspan="2">215.01 ± 20.88</td><td>219.91 ± 22.33</td><td>215.95 ± 21.58</td><td>220.64 ± 22.95</td><td>215.95 ± 22.36</td>

<td>12</td><td colspan="2">387.98 ± 45.76</td><td>284.09 ± 32.64</td><td>255.85 ± 34.44</td><td>181.40 ± 21.15</td><td>379.15 ± 46.00</td>

<td>16</td><td>828.95 58.76</td><td>±</td><td>393.95 ± 42.09</td><td>268.23 ± 47.77</td><td>180.18 ± 26.25</td><td>737.84 ± 80.06</td>

<td>19</td><td>1425.22 ± 101.9</td><td></td><td>514.88 ± 55.57</td><td>346.01 ± 62.50</td><td>207.28 ± 42.54</td><td>1195.5 ± 67.91</td>

<td>23</td><td>2169.9 170.8</td><td>±</td><td>670.36 ± 54.19</td><td>373.01 ± 63.35</td><td>232.25 ± 37.11</td><td>1702.5 ± 101.8</td>

Table 12. Tumor sizes in the Different Treatment Groups on the HCC827 model

<td colspan="5">Tumor volume (mm³)^and</td>

<td>Days</td><td>Substrate, PO, QD Subsoil</td><td>Compound 3, PO, QD 50mpk (PEG)</td><td>Compound 3, PO, QD 50mpk (0.5% MC)</td><td>Gefitinib, PO, QD 100mpk</td>

<td>14</td><td>215.94 ± 25.70</td><td>211.90 ± 23.00</td><td>211.14 ± 25.11</td><td>212.28 ± 26.35</td>

<td>18</td><td>291.15 ± 24.42</td><td>188.72 ± 28.03</td><td>216.63 ± 27.69</td><td>59.55 ± 25.20</td>

<td>21</td><td>353.24 ± 25.64</td><td>136.96 ± 16.40</td><td>245.14 ± 33.44</td><td>4.61 ± 3.16</td>

<td>25</td><td>453.43 ± 24.72</td><td>95.73 ± 15.38</td><td>216.42 ± 28.06</td><td>1.25 ± 1.25</td>

<td>28</td><td>519.39 ± 22.26</td><td>111.96 ± 22.05</td><td>231.08 ± 30.81</td><td>1.25 ± 1.25</td>

<td>32</td><td>638.78 ± 32.70</td><td>82.28 ± 24.08</td><td>277.59 ± 42.02</td><td>1.25 ± 1.25</td>

<td>35</td><td>762.43 ± 47.22</td><td>67.63 ± 24.22</td><td>293.64 ± 43.98</td><td>1.88 ± 1.32</td>

<td>39</td><td>1092.53 ± 99.28</td><td>69.44 ± 30.35</td><td>328.53 ± 43.51</td><td>1.88 ± 1.32</td>

<td>42</td><td>1324.76 ± 141.54</td><td>79.71 ± 28.86</td><td>302.31 ± 35.83</td><td>10.95 ± 6.13</td>

<td>46</td><td>1736.94 ± 217.03</td><td>84.26 ± 35.62</td><td>284.44 ± 27.00</td><td>23.71 ± 11.84</td>

<td>49</td><td>1920.11 ± 256.36</td><td>77.59 ± 42.07</td><td>299.28 ± 31.79</td><td>41.00 ± 20.52</td>

Table 13. Tumor sizes in the Different Treatment Groups on the A431 model

<td colspan="4">Tumor volume (mm³)^and</td>

<td>Days</td><td>Substrate, PO, QD</td><td>Compound 3, PO, QD 100mpk</td><td>Gefitinib, PO, QD 100mpk</td>

<td>11</td><td>241.34 ± 28.69</td><td>240.95 ± 26.46</td><td>239.83 ± 23.30</td>

<td>14</td><td>472.09 ± 71.50</td><td>399.68 ± 42.62</td><td>203.74 ± 22.97</td>

<td>18</td><td>860.82 ± 120.62</td><td>867.62 ± 70.54</td><td>139.70 ± 26.94</td>

<td>21</td><td>1211.0 ± 157.77</td><td>1166.1 ± 94.08</td><td>139.70 ± 22.07</td>

<td>25</td><td>1666.6 ± 233.36</td><td>1627.7 ± 146.0</td><td>154.79 ± 32.62</td>

<td>Attention:</td><td>a. Medium ± SEM</td><td></td><td></td>

Inhibition of Tumor Growth [03861 Inhibition of tumor growth on the NCI-H1975, HCC827 and A431 models is summarized in Table 14, Table 15, and Table 16, respectively.

<td colspan="5">Table14. The Effect of Compounds in the Treatment of H1975 Tumor Heteroplastoma</td>

<td>Treatment</td><td>Tumor size (mm³)^and in on 23 (14)</td><td>T / C (%)</td><td>TC (days) at 300mm³</td><td>^p</td>

<td>Subsoil</td><td>2170 ± 171</td><td></td><td>-</td><td>-</td>

<td>Compound 3</td><td></td><td></td><td></td><td>0.000</td>

<td>25 mg / kg after qd</td><td>670 ± 54</td><td>28.5%</td><td>2.11</td><td></td>

<td>Compound 3</td><td></td><td></td><td></td><td>0.000</td>

<td>50 mg / kg after qd</td><td>373 ± 63</td><td>16.0%</td><td>6.76</td><td></td>

<td>Compound 3</td><td></td><td></td><td></td><td>0.000</td>

<td>100 mg / kg after qd</td><td>232 ± 37</td><td>9.9%</td><td>&gt; 14</td><td></td>

<td>gefitinib</td><td></td><td></td><td></td><td>0.345</td>

<td>100 mg / kg after qd</td><td>1702 ± 102</td><td>77.4%</td><td>0.08</td><td></td>

Table 15. Effect of Compounds in the Treatment of Tumor HCV tumor Model HCC827

<td>Treatment</td><td>Size (mm³)^and in 49 (35)</td><td>tumor day</td><td>T / C (%)</td><td>TRR^b in on day 49 (35)</td><td>TC (days) at 300mm³</td><td>P</td>

<td>Subsoil</td><td>1920 ± 256</td><td></td><td>-</td><td>-</td><td>-</td><td>-</td>

<td>Compound 3 (PEG)</td><td></td><td></td><td></td><td></td><td></td><td>0.001</td>

<td>50 mg / kg after qd</td><td>78 ± 42</td><td></td><td>2.8%</td><td>71.4%</td><td>&gt; 35</td><td></td>

<td>Compound 3 (MC)</td><td></td><td></td><td></td><td></td><td></td><td>0.002</td>

<td>50 mg / kg after qd</td><td>299 ± 32</td><td></td><td>14.3%</td><td>-45.0%</td><td>17.3</td><td></td>

<td>gefitinib</td><td></td><td></td><td></td><td></td><td></td><td>0.001</td>

<td>100 mg / kg after qd</td><td>41 ± 21</td><td></td><td>2.4%</td><td>75.7%</td><td>&gt; 35</td><td></td>

Table 16. Effect of Compounds in the Treatment of Tumor A431 Hematoplant

<td>Treatment</td><td>Tumor size (mm³)^and in on day 25 (14)</td><td>T / C (%)</td><td>TC (days) 300mm³</td><td>at p</td>

<td>Subsoil</td><td>1667 ± 233</td><td></td><td>-</td><td>-</td>

<td>Compound 3</td><td></td><td></td><td></td><td>0.999</td>

<td>100 mg / kg after qd</td><td>1628 ± 146</td><td>98.3%</td><td>0.35</td><td></td>

<td>gefitinib</td><td></td><td></td><td></td><td>0.001</td>

<td colspan="2">Treatment Tumor size (mm³)^and in T / C</td><td rowspan="2">TC (days) at p 300mm³</td>

<td>on day 25 (14)</td><td>(%)</td>

<td>100 mg / kg after qd 154 ± 33</td><td>8.7%</td><td>&gt; 14</td>

<td>Note: a. Medium ± SEM</td><td></td><td></td>

<td colspan="3">b. Tumor regression rate (%) = (1-volume tumor after treatment / tumor volume from initial treatment) * 100%</td>

Tumor Growth Curve [0387] The tumor growth curve in various groups of tumor-bearing mice on models NCI-H1975, HCC827 and A431 is shown in Figure 6, Figure 7 and Figure 8, respectively.

Tumor Mass [0388] The tumor masses of mice in different groups on models NCI-H1975, HCC827 and A431 are shown in Table 17, Table 18, and Table 19, respectively.

<td colspan="4">Table 17. Antitumour activity of Compounds in the treatment of the NCI-H1975 Model</td>

<td>Treatment</td><td>Mass of Guz (g) on ​​day 23 (14)</td><td>IR ^and on 23 (14)</td><td>^p</td>

<td>Subsoil</td><td>1.99 ± 0.16</td><td>-</td><td>-</td>

<td>Compound 3 25 mg / kg after qd</td><td>0.70 ± 0.04</td><td>65.0%</td><td>0.001</td>

<td>Compound 3 50 mg / kg after qd</td><td>0.34 ± 0.08</td><td>82.8%</td><td>0.000</td>

<td>Gefitinib 100 mg / kg after qd</td><td>1.69 ± 0.11</td><td>15.1%</td><td>0.717</td>

Table 18. Antibiotic activity of Compounds in the treatment of Model HCC827

<td>Treatment</td><td>The weight of the tumor (g) on ​​day 49 (35)</td><td>IR ^and on day 49 (35) p</td>

<td>Subsoil</td><td>1.94 ± 0.32</td><td>- -</td>

<td>Compound 3 (PEG) 50 mg / kg after qd</td><td>0.06 ± 0.02</td><td>96.7% .004</td>

<td>Compound 3 (MC) 50 mg / kg after qd</td><td>0.26 ± 0.04</td><td>86.5% .008</td>

<td>Gefitinib 100 mg / kg after qd</td><td>0.03 ± 0.02</td><td>98.3% .004</td>

Table 19. Antibiotic activity of Compounds in the treatment of A431 Model

Treatment of tumor mass (g) on ​​IR ^and on 23 (14) p

23 (14)

Subsoil

1.67 ± 0.29

100

<td>Treatment</td><td>Tumor mass (g) on 23 (14)</td><td>IR ^and on 23 (14)</td><td>^p</td>

<td>Compound 3 100 mg / kg after qd</td><td>1.60 ± 0.22</td><td>4.0%</td><td>0817</td>

<td>Gefitinib 100 mg / kg after qd</td><td>0.13 ± 0.03</td><td>92.5%</td><td>0.000</td>

<td>a: IR (Braking rate) =</td><td colspan="2">(TWKontrola - TWLeczenie) / TWKontrol ^x 100%</td><td></td>

Biological Example D

Synthesis of Maleate Salt or Compound 3 Hydrochloride Salt and Pharmacokinetic Study [0389] Synthesis of maleate salt and hydrochloride salt from the free base of Compound 3 is shown

<img file="PL2880035T3_D0083.tif" />

[0390] To the free base of Compound 3 (2 g) in ethanol / water (5:95, 22 mL) at 40 ° C was added dropwise maleic acid (1.2 eq.) Or HCl (2.2 eq.). After dissolving the solid, the solution was cooled to room temperature, and left to stand overnight. The resulting crystals (light yellow or off-white) were collected, washed with cold water and dried overnight (over 85% yield).

Pharmacokinetic studies in rats with Compound 3 in free base forms, maleate salts and HCl salts:

[0391] A PK pharmacokinetic comparison study was performed on female rats (Zhe Jiang AMS) using Compound 3 in the free base forms, the maleate salt and the HCl salt. The detailed test conditions together with the experimental results are shown in Table 20 below:

Table 20

<td>Relationship</td><td>formulation</td><td>Dose (Mg / kg)</td><td>Road</td><td>F%</td><td>AUClast (Ng / mL * h)</td><td>t ½ (h)</td><td>Cmax (po) or C0 (iv) (ng / ml)</td>

<td>Free</td><td>PEG200: D5W</td><td>4.7</td><td>iv</td><td>on</td><td>2195+</td><td>1.6 ± 0.</td><td>4102.1 ±</td>

<td>principle of the Union 3</td><td>(50:50, v / v)</td><td></td><td></td><td></td><td>140.2</td><td>04</td><td>675.8</td>

<td>Free</td><td>0.5% MC</td><td>30.56</td><td>after</td><td>10.7 ±</td><td>1526.8 ±</td><td>2.4 ± 0.</td><td>242.0 ±</td>

<td>principle of the Union 3</td><td></td><td></td><td></td><td>2.6</td><td>364.6</td><td>6</td><td>37.6</td>

<td>HCl salt</td><td>0.5% MC</td><td>40.88</td><td>p.^about.</td><td>30.6 ±</td><td>5853.8 ±</td><td>2.4 ± 1.</td><td>1259.3 ±</td>

<td>Relationship 3</td><td></td><td></td><td></td><td>8.2</td><td>1565.4</td><td>0</td><td>359.0</td>

101

<td>Relationship</td><td>formulation</td><td>Dose (Mg / kg)</td><td>Road</td><td>F%</td><td>AUClast (Ng / mL * h)</td><td>t ½ (h)</td><td>Cmax (po) or C0 (iv) (ng / ml)</td>

<td>salt</td><td>0.5% MC</td><td>42.00</td><td>after</td><td>32.7 ±</td><td>6412.2 ±</td><td>2.3 ± 0.</td><td>1540.0 ±</td>

<td>Maleinian of the Union 3</td><td></td><td></td><td></td><td>9.8</td><td>1917.8</td><td>4</td><td>528.5</td>

<td>Free</td><td>PEG200: D5W</td><td>37.6</td><td>after</td><td>23.0 ±</td><td>4041.0 ±</td><td>2.8 ± 0.</td><td>1263.3 ±</td>

<td>principle of the Union 3</td><td>(50:50, v / v)</td><td></td><td></td><td>10.8</td><td>1892.3</td><td>3</td><td>270.2</td>

[0392] The vnikises show that in the same 0.5% methyllcellulose (MC) formulation, the salt forms (both maleate salt and HCl salt) had about 3-fold better bioavailability compared to the free base.

Biological Example E [0393] Compound 3 was tested against many other protein kinases.

[0394] Test Description. In vitro profiling of protein kinases was performed using the "HotSpot" test platforms. Briefly, the specific parv kinase / substrate together with the vaginal co-factors were resolved in the reaction buffer. Compounds were provided for the reaction, after 15-20 minutes later a mixture of ATP (Sigma, St. Louis MO) and³³P ATP (Perkin Elmer, Waltham MA) to a final concentration of 10 μΜ. The reactions were carried out at room temperature for 120 min, and then the reaction mixtures were applied to an ion-exchange P81 filter paper (Whatman Inc., Piscatawav, NJ). Unbound phosphate was removed by intense washing of the paper in 0.75% phosphoric acid. After subtracting the background from the control reactions containing the inactive enzyme, kinase activity data was expressed as a percentage of the remaining kinase activity in the test samples compared to the substrate reaction (dimethoxysulfoxide). IC 50 values ​​and curve fitting were obtained using Prism (GraphPad Software).

[0395] Reaction Conditions: Buffer Conditions: 20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg / mL BSA, 0.1 mM Na3VO4, 2 mM DTT, and 1% DMSO. ATP concentration: 10 μΜ. Reaction Time: 2 hours. Compound 3 was tested at 10 concentrations in the following manner compared to the common reference compound kinase inhibitor staurosporine. Compounds were tested in a 10-point trvba IC 50 with a 3-fold serial dilution ranging from 10 μM. The control compound was tested in a 10-point IC 50 with a 3-fold serial dilution ranging from 20 μM. All kinase reactions were performed in the presence of ATP. The test concentrations are given below in molar units (M) ._

<td>Compound 3</td><td>Staurosporine-control</td>

<td>1.00E-05</td><td>2.00-05</td>

<td>3.33-06</td><td>6.67-06</td>

<td>1.11-06</td><td>2.22-06</td>

102

<td>Compound 3</td><td>Staurosporine-control</td>

<td>3.70-07</td><td>7.41-07</td>

<td>1.23-07</td><td>2.47-07</td>

<td>4.12-08</td><td>8.23-08</td>

<td>1.37-08</td><td>2.74-08</td>

<td>4.57-09</td><td>9.14-09</td>

<td>1.52-09</td><td>3.05-09</td>

<td>5.08-10</td><td>1.02-09</td>

[0396] The following kinase enzymes were tested:

103

<td>Name</td><td>Symbol HUGO</td><td>General substratum</td><td>Genbank Access</td><td>No. access proteins</td><td>Maple</td><td>Mutation</td><td>Expression</td><td>tag</td>

<td>BLK</td><td>BLK</td><td>pEY</td><td>NP_001706</td><td>P51451</td><td>full length</td><td>-</td><td>Insect</td><td>N-terminal His</td>

<td>BMX / ETK</td><td>BMX</td><td>pEY</td><td>NP 001712</td><td>P51813</td><td>full length</td><td>-</td><td>baculovirus in Sf21 insect cells</td><td>C-terminal His</td>

<td>BTK</td><td>BTK</td><td>pEY</td><td>NP_000052</td><td>Q06187</td><td>full length</td><td>-</td><td>Insect</td><td>N-terminal His6tagowana</td>

<td>ITK</td><td>ITK</td><td>MBP</td><td>NP_005537</td><td>Q08881</td><td>full length</td><td>-</td><td>Insect</td><td>N-terminal GST</td>

<td>JAK2</td><td>JAK2</td><td>pEY</td><td>NP_004963</td><td>060674</td><td>aa 8091132 + g</td><td>-</td><td>Insect</td><td>N-terminal GST</td>

<td>JAK3</td><td>JAK3</td><td>JAK3tide</td><td>NP_000206</td><td>P52333</td><td>aa 7811124</td><td>-</td><td>Insect</td><td>N-terminal GST</td>

<td>TEC</td><td>TEC</td><td>pEY + Mn</td><td>NP_003206</td><td>P42680</td><td>full length</td><td>-</td><td>insect cell with the system baculovirus</td><td>C-terminal His-tag</td>

<td>TXK</td><td>TXK</td><td>ABLtide</td><td>NP_003319.2</td><td>P42681</td><td>full length</td><td>-</td><td>Insect</td><td>N-terminal GST-tag</td>

104

<td>Name</td><td>Symbol HUGO</td><td>General substratum</td><td>Access Number Genbank</td><td>No. access proteins</td><td>Maple</td><td>Mutation</td><td>Expression</td><td>tag</td>

<td>FLT3 (D835Y)</td><td>FLT3</td><td>Abltide</td><td>NP_004110</td><td>P36888</td><td>aa 564958</td><td>D835Y</td><td>insect cell with system baculovirus</td><td>C-terminal His 6-tag</td>

[0397] Results: The IC 50 values ​​for each of the kinase targets can be summarized as follows:

<td></td><td colspan="2">IC50 Compound (M)</td>

<td>kinases</td><td>Compound 3</td><td>staurosporine</td>

<td>BLK</td><td>2.34-10</td><td>1.22-09</td>

<td>BMX / ETK</td><td>3.45-10</td><td>2.19-09</td>

<td>BTK</td><td>3.99-10</td><td>5.78-09</td>

<td>FLT3 (D835Y)</td><td>1.09-08</td><td>1.63-11</td>

<td>ITK</td><td>5.17-10</td><td>8.96-09</td>

<td>JAK2</td><td>5.01-07</td><td>&lt;1.00E-9</td>

<td>JAK3</td><td>9.11-11</td><td>2.99-11</td>

<td>TEC</td><td>6.69-10</td><td>2.68-08</td>

<td>TXK</td><td>6.98-10</td><td>1.28-08</td>

[0398] The following lists IC50 values ​​and raw data for each target enzyme.

[0399] The percentage of activity is a relative value as compared to only the buffer solution. DMSO is listed as a reference. The IC50 value for Compound 3 for BLK was 0.23 nM. Suitable curves for Compound 3 are shown in Figures 15A and 15B, respectively.

<td colspan="4">% Activities</td>

<td>Conc. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>1.00E-05</td><td>-0.41</td><td>0.57</td><td>2.00-05</td>

<td>3.33-06</td><td>2.22</td><td>2.47</td><td>6.67-06</td>

<td>1.11-06</td><td>4.39</td><td>1.46</td><td>2.22-06</td>

<td>3.70-07</td><td>1.49</td><td>-1.03</td><td>7.41-07</td>

<td>1.23-07</td><td>-0.50</td><td>-2.23</td><td>2.47-07</td>

<td>4.12-08</td><td>-3.83</td><td>-1.99</td><td>8.23-08</td>

<td>1.37-08</td><td>-0.37</td><td>1.34</td><td>2.74-08</td>

<td>4.57-09</td><td>1.07</td><td>10.32</td><td>9.14-09</td>

<td>1.52-09</td><td>14.53</td><td>27.25</td><td>3.05-09</td>

<td>5.08-10</td><td>29.73</td><td>53.08</td><td>1.02-09</td>

<td>DMSO</td><td>102.02</td><td>97.98</td><td>DMSO</td>

[0400] The IC50 for Compound 3 for BMX / ETK was 0.35 nM. The curve for Compound 3 is shown in Figure 16._

<td colspan="4">% Activities</td>

<td>Conc. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>1.00E-05</td><td>1.44</td><td>9.22</td><td>2.00-05</td>

<td>3.33-06</td><td>-4.22</td><td>-4.43</td><td>6.67-06</td>

<td>1.11-06</td><td>-0.82</td><td>-1.61</td><td>2.22-06</td>

<td>3.70-07</td><td>-3.51</td><td>-0.71</td><td>7.41-07</td>

<td>1.23-07</td><td>-0.30</td><td>1.44</td><td>2.47-07</td>

<td>4.12-08</td><td>1.70</td><td>8.90</td><td>8.23-08</td>

<td>1.37-08</td><td>1.03</td><td>18.68</td><td>2.74-08</td>

<td>4.57-09</td><td>7.02</td><td>24.52</td><td>9.14-09</td>

<td>1.52-09</td><td>9.87</td><td>39.02</td><td>3.05-09</td>

<td>5.08-10</td><td>37.60</td><td>64.45</td><td>1.02-09</td>

<td>DMSO</td><td>98.60</td><td>96.41</td><td>DMSO</td>

[0401] The IC 50 for Compound 3 for BTK was 0.40 nM. The curve for Compound 3 is shown in Figure 17. _

<td colspan="4">% Activities</td>

<td>Conc. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>1.00E-05</td><td>3.65</td><td>1.93</td><td>2.00-05</td>

<td>3.33-06</td><td>0.69</td><td>-0.14</td><td>6.67-06</td>

<td>1.11-06</td><td>-0.26</td><td>0.21</td><td>2.22-06</td>

<td>3.70-07</td><td>3.46</td><td>3.51</td><td>7.41-07</td>

<td>1.23-07</td><td>1.71</td><td>6.52</td><td>2.47-07</td>

<td>4.12-08</td><td>1.89</td><td>10.70</td><td>8.23-08</td>

<td>1.37-08</td><td>4.18</td><td>20.50</td><td>2.74-08</td>

<td>4.57-09</td><td>3.32</td><td>41.55</td><td>9.14-09</td>

<td>1.52-09</td><td>11.43</td><td>60.75</td><td>3.05-09</td>

<td>5.08-10</td><td>40.24</td><td>81.36</td><td>1.02-09</td>

<td>DMSO</td><td>96.43</td><td>98.79</td><td>DMSO</td>

[04021 IC50 for Compound 3 for FLT3 (D835Y) was 10.90 nM. The curve for Compound 3 is shown in Figure 18._

<td colspan="4">% Activities</td>

<td>Conc. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>1.00E-07</td><td>11.32</td><td>-1.65</td><td>1.00E-07</td>

<td>3.33-08</td><td>27.78</td><td>0.98</td><td>3.33-08</td>

<td>1.11-08</td><td>49.45</td><td>0.31</td><td>1.11-08</td>

<td>3.70-09</td><td>70.12</td><td>2.34</td><td>3.70-09</td>

<td>1.23-09</td><td>92.41</td><td>-0.82</td><td>1.23-09</td>

<td>4.12-10</td><td>99.20</td><td>3.17</td><td>4.12-10</td>

<td>1.37-10</td><td>104.37</td><td>2.10</td><td>1.37-10</td>

<td>4.57-11</td><td>94.42</td><td>20.65</td><td>4.57-11</td>

<td>1.52-11</td><td>97.21</td><td>53.79</td><td>1.52-11</td>

<td>5.08-12</td><td>102.50</td><td>85.41</td><td>5.08-12</td>

<td>DMSO</td><td>99.21</td><td>100.79</td><td>DMSO</td>

[04031 The IC 50 for Compound 3 for ITK was 0.52 nM. The curve for Compound 3 is shown in Figure 19. _

<td colspan="4">% Activities</td>

<td>Conc. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>1.00E-05</td><td>0.79</td><td>0.35</td><td>2.00-05</td>

<td>3.33-06</td><td>0.01</td><td>0.60</td><td>6.67-06</td>

<td>1.11-06</td><td>1.27</td><td>-0.12</td><td>2.22-06</td>

<td>3.70-07</td><td>1.16</td><td>1.90</td><td>7.41-07</td>

<td>1.23-07</td><td>0.90</td><td>4.54</td><td>2.47-07</td>

<td>4.12-08</td><td>1.70</td><td>9.04</td><td>8.23-08</td>

<td>1.37-08</td><td>1.34</td><td>19.24</td><td>2.74-08</td>

<td>4.57-09</td><td>2.39</td><td>49.31</td><td>9.14-09</td>

<td>1.52-09</td><td>4.41</td><td>76.22</td><td>3.05-09</td>

<td>5.08-10</td><td>52.22</td><td>95.37</td><td>1.02-09</td>

<td>DMSO</td><td>101.99</td><td>97.19</td><td>DMSO</td>

108 [0404] The IC 50 for Compound 3 for JAK2 was 501 nM. The curve for Compound 3 is shown in Figure 20. _

<td colspan="4">% Activities</td>

<td>Conc. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>1.00E-05</td><td>-2.83</td><td>-5.43</td><td>2.00-05</td>

<td>3.33-06</td><td>9.17</td><td>-8.65</td><td>6.67-06</td>

<td>1.11-06</td><td>23.15</td><td>-6.44</td><td>2.22-06</td>

<td>3.70-07</td><td>60.18</td><td>-6.42</td><td>7.41-07</td>

<td>1.23-07</td><td>94.11</td><td>-6.76</td><td>2.47-07</td>

<td>4.12-08</td><td>100.48</td><td>-7.25</td><td>8.23-08</td>

<td>1.37-08</td><td>99.88</td><td>-3.41</td><td>2.74-08</td>

<td>4.57-09</td><td>98.95</td><td>-0.60</td><td>9.14-09</td>

<td>1.52-09</td><td>103.16</td><td>5.51</td><td>3.05-09</td>

<td>5.08-10</td><td>98.40</td><td>18.77</td><td>1.02-09</td>

<td>DMSO</td><td>103.16</td><td>100.65</td><td>DMSO</td>

[0405] To determine JAK3 inhibition, Compound 3 was assessed as in other assays, but starting at a concentration of 100 nM. The dilution ratio was 3-fold as in the remaining tests, obtaining a concentration range in the 100nM test to 5.08 pM. The IC50 for Compound 3 for JAK3 was 91.1 pM. The curve for Compound 3 is shown in Figure 21.

<td colspan="4">% Activities</td>

<td>Conc. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>1.00E-07</td><td>-0.21</td><td>-0.79</td><td>1.00E-07</td>

<td>3.33-08</td><td>0.22</td><td>-0.88</td><td>3.33-08</td>

<td>1.11-08</td><td>-1.35</td><td>-0.51</td><td>1.11-08</td>

<td>3.70-09</td><td>-0.10</td><td>0.09</td><td>3.70-09</td>

<td>1.23-09</td><td>1.25</td><td>-1.86</td><td>1.23-09</td>

<td>4.12-10</td><td>9.43</td><td>0.36</td><td>4.12-10</td>

<td>1.37-10</td><td>35.13</td><td>4.48</td><td>1.37-10</td>

<td>4.57-11</td><td>72.27</td><td>41.45</td><td>4.57-11</td>

<td>1.52-11</td><td>89.22</td><td>70.04</td><td>1.52-11</td>

<td>5.08-12</td><td>92.29</td><td>88.93</td><td>5.08-12</td>

109

<td colspan="4">% Activities</td>

<td>Conc. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>DMSO</td><td>102.06</td><td>102.66</td><td>DMSO</td>

[0406] The IC 50 for Compound 3 for TEC was 0.67 nM. The curve for Compound 3 is shown in Figure 22. _

<td colspan="4">% Activities</td>

<td>Conc. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>1.00E-05</td><td>2.69</td><td>1.43</td><td>2.00-05</td>

<td>3.33-06</td><td>7.10</td><td>3.40</td><td>6.67-06</td>

<td>1.11-06</td><td>5.72</td><td>3.35</td><td>2.22-06</td>

<td>3.70-07</td><td>11.05</td><td>8.71</td><td>7.41-07</td>

<td>1.23-07</td><td>14.88</td><td>18.31</td><td>2.47-07</td>

<td>4.12-08</td><td>19.39</td><td>31.27</td><td>8.23-08</td>

<td>1.37-08</td><td>20.14</td><td>47.73</td><td>2.74-08</td>

<td>4.57-09</td><td>22.53</td><td>70.74</td><td>9.14-09</td>

<td>1.52-09</td><td>32.07</td><td>79.41</td><td>3.05-09</td>

<td>5.08-10</td><td>72.24</td><td>93.91</td><td>1.02-09</td>

<td>DMSO</td><td>99.73</td><td>99.19</td><td>DMSO</td>

[0407] The IC50 for Compound 3 for TXK was 0.70 nM. The curve for Compound 3 is shown in Figure 23. _

<td colspan="4">% Activities</td>

<td>St. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>1.00E-05</td><td>-0.05</td><td>-0.60</td><td>2.00-05</td>

<td>3.33-06</td><td>-1.39</td><td>-0.74</td><td>6.67-06</td>

<td>1.11-06</td><td>4.24</td><td>2.69</td><td>2.22-06</td>

<td>3.70-07</td><td>-2.01</td><td>1.66</td><td>7.41-07</td>

<td>1.23-07</td><td>3.26</td><td>9.25</td><td>2.47-07</td>

<td>4.12-08</td><td>2.76</td><td>17.37</td><td>8.23-08</td>

<td>1.37-08</td><td>0.31</td><td>32.52</td><td>2.74-08</td>

<td>4.57-09</td><td>4.11</td><td>58.10</td><td>9.14-09</td>

<td>1.52-09</td><td>23.68</td><td>79.12</td><td>3.05-09</td>

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<td colspan="4">% Activities</td>

<td>St. (M)</td><td>Compound 3</td><td>staurosporine</td><td>Conc. Stauro (M)</td>

<td>5.08-10</td><td>58.85</td><td>95.83</td><td>1.02-09</td>

<td>DMSO</td><td>96.09</td><td>100.00</td><td>DMSO</td>

Data for other compounds

1. ELISA test (EGFR):

[0408] Using the protocol described in biological example B-2, the following compounds were also tested by ELISA (EGFR). Wvniki are shown in Table 21 below. Table 21

ELISA test (EGFR)

<td>ZWK / EGFR</td><td>IC50 (uM) T790M / L858R EGF stimulation)</td><td>H1975 (lack</td><td>IC50 (uM) A431 WT (EGF simulation)</td><td>selectivity A431 / H1975</td>

<td>Zwk 1</td><td>.0063</td><td></td><td>.6470</td><td>102.70</td>

<td>Zwk 2</td><td>0.0010</td><td></td><td>.3000</td><td>300.00</td>

<td>Zwk 4</td><td>.0670</td><td></td><td>10.9000</td><td>162.69</td>

<td>Zwk 5</td><td>.2430</td><td></td><td>27.5000</td><td>113.17</td>

<td>Zwk 6</td><td>.0398</td><td></td><td>9.2700</td><td>232.91</td>

<td>Zwk 7</td><td>12.4100</td><td></td><td>&gt; 20</td><td>ON</td>

<td>Zwk 34</td><td>.0217</td><td></td><td>2.0170</td><td>92.95</td>

2. Other kinases - Testv Enzvmatvczne [0409] Using the protocol described in biological example E, the following compounds were also tested against other kinases (BTK, Jak1, Jak2 and Jak3) (Table 22 below).

Table 22

<td rowspan="2">Zwk \ Kinases</td><td colspan="4">IC50 Enzymatic Tests</td>

<td>BTK</td><td>JAK3</td><td>JAK2</td><td>JAK1</td>

<td>Zwk 1</td><td>5.79-10</td><td>3.66-11</td><td>5.70-07</td><td>3.22-06</td>

<td>Zwk 2</td><td>3.62-10</td><td>3.65-11</td><td>2.06-06</td><td>1.40-05</td>

<td>Zwk 3</td><td>3.99-10</td><td>9.11-11</td><td>5.01-07</td><td>3.27-06</td>

<td>Staurosporine control</td><td>4.97-09</td><td>1.92-11</td><td>1.96-10</td><td>3.65-10</td>

Other kinases (BTK and JAK3) - Cell-based tests

[0410] The protocol for the JAK3 and BTK ClariCELL ™ tests is described below:

Human embryonic kidney HEK293 cells were transiently transfected with either wt human BTK, human wt JAK3, or human JAK3 with dead kinase (for JAK3 negative controls). For BTK negative controls, BTK plus wt transfection was used plus 1 IM Ibrutinib treatment.

• Cells were measured in 96-well plates at approximately 8,000 cells / well for BTK, or 20,000 cells per well for JAK3.

• Eight three-fold serial dilutions (or two times for tofacitinib) were prepared for each compound in 100% DMSO.

The compounds were then diluted in water to 10x the final concentration in the assay and 6% DMSO.

• The compounds were added to the cells in 96-well plates (10-fold dilution in tissue culture medium) for a final concentration of 1x compound and 0.6% DMSO.

• The cells were incubated with the compound at 37 ° C for 2 hours.

The cells were lysed, and the lysate was transferred to an ELISA plate that was previously coated with the antibody to capture the substrate (either human BTK or human JAK3).

• The plates were washed, then incubated with the antibody to detect complete tyrosine phosphorylation *.

• The plates were washed then incubated with a secondary antibody labeled with HRP.

• HRP substrate was added and absorbance read at 450nm.

* It should be noted that since the pan-anti-phosphotyrosine antibody was used, it is possible that tyrosine kinase activities may be measured in addition to BTK or JAK3.

[0411] Test results for cell-based BTK assays are set forth in Table 23 below.

Table 23

<td rowspan="2">Zwk \ Kinases</td><td>Cell-based tests (CAI) ΚΑίμΥΙ)</td>

<td>BTK</td>

<td>Zwk 2</td><td>0012 0.059</td>

<td></td><td>0.026</td>

<td>Zwk 3</td><td>0.036</td>

<td></td><td>0.059</td>

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<td rowspan="2">Zwk \ Kinases</td><td>Cell-based tests (CAI) Κ50 (μΜ)</td>

<td>BTK</td>

<td>Control (Ibrutinib)</td><td>0.013 0.013 0.067 0012</td>

[0412] Test results for cell-based JAK3 assays are set forth in the following Table 24.

Table 24

<td rowspan="2">Zwk \ Kinases</td><td>Cell-based tests (CAI) Κ50 (μΜ)</td>

<td>JAK3</td>

<td>Zwk 2</td><td>0.20 0.36</td>

<td>Zwk 3</td><td>0.23 0.28 0.25 0.36</td>

<td>Control (Tofacitinib)</td><td>2.3 2.3 1.2 3.7</td>

Dorota Rzążewska

Patent Attorney

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