COMBINATION THERAPIES

FOR THE TREATMENT OF BRAIN CANCER

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority to US Provisional Application 63/423,649, filed November 8, 2022, the entire contents of which are incorporated by reference herein.

BACKGROUND

Glioblastomas (GBM) are aggressive and universally lethal brain tumors. Existing treatments are based on a combination of aggressive therapies. Concomitant chemotherapy and radiotherapy are generally implemented after surgical tumor resection. However, this combination of surgery, chemo and radiotherapy only offers a median survival of less than 15 months. A notable feature of GBM tumors is that they are exquisitely resistant to therapy- induced cell death, which may in part explain this dismal survival time. With only 3 - 5% of patients surviving longer than 3 years, due to disease recurrence, GBM remains incurable. Accordingly, there is an ongoing, unmet need for new methods for treating GBM.

SUMMARY OF THE INVENTION

In one aspect, the present disclosure provides methods treating brain cancer in a subject in need thereof, comprising administering an inhibitor of synaptic signaling to the subject.

In aother aspect, the present disclosure provides methods of treating brain cancer in a subject in need thereof, comprising conjointly administering an inhibitor of synaptic signaling and a first EGFR inhibitor to the subject.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A shows tumor characteristics, newly diagnosed vs recurrent, tumor grade of patient tumors, EGFRvIII splice variant, MGMT status and GBM subtypes which were determined using bulk RNA sequencing data with previously described gene sets. Whole exome sequencing describing copy number gains and losses and single nucleotide variant (SNV) mutations (somatic or germline depending on sample availability) across the most frequently recurrently altered genes in GBM as described by the TCGA (Brennan et al., 2013). BH3 of the primed state, heatmaps represent percent of cytochrome c release with each peptide, assessed with flow cytometry, relative to DMSO treated control. FIG. IB shows BH3 profiling of patient samples grouped by tumor grade. FIG. 1C shows the time to progressions for UCLA glioma patients, grouped by tumor grade. FIG. ID shows BH3 profiling of the same patient tumor at first and second recurrence.

FIG. 2A shows the BH3 profiling of freshly isolated patient samples and gliomaspheres grown in cell culture. FIG. 2B shows BH3 profiling of purified orthotopic xenografts relative to their matched gliomaspheres. FIG. 2C shows BH3 profiling of purified subcutaneous xenografts relative to their matched orthotopic xenografts. FIG. 2D shows the Concentration of Compound 1 in mouse brain and subcutaneous tumor tissue. FIG. 2E shows the tumor burden of orthotopic or subcutaneous xenografts treated with either vehicle or Compound 1. Mice treated with Compound 1 had a lower tumor burden vs. vehicle. FIG. 2F shows the differential gene expression and subsequent gene ontology of genes with high expression in orthotopic xenografts and low expression in subcutaneous environments, relative to patient tumors.

FIG. 3A shows immunofluorescent image of gliomaspheres in co-culture with neurons. FIG. 3B shows BH3 profiling of gliomaspheres GS025, cultured in neuron conditioned media or in direct co-culture with the neurons. FIG. 3C shows BH3 profiling of GS025 co-culture with neurons with the synaptic inhibitors TTX and NBQX. FIG. 3D shows BH3 profiling of GS025 co-culture with neurons with the gap junction inhibitor meclofenamate. FIG. 3E shows the cell viability of gliomasphere GS025 treated with either TMZ or Compound 1 with neuron co-culture. FIG. 3F shows the cell viability of gliomasphere GS025 treated with either Compound 1 with neuron co-culture in combination with either TTX or NBQX.

FIG. 4A shows CREB target gene score of patient samples. Gliomaspheres, sphere- derived-xenografts and patient-derived-xenografts. FIG 4B shows an immunoblot and BH3 profiling of GBM tumor cells co-cultured with neurons and treated with the CREB inhibitor, 666-15. FIG. 4C shows the gene expression of the BCL2 protein family across environments. FIG. 4D shows an immunoblot of GBM tumor cells co-cultured with neurons. Quantification in graph is of the protein BIM. FIG. 4E shows an immunoblot of GBM tumor cells co-cultured with neurons and treated with TTX.

FIGS. 5A & 5B show the ex- vivo dynamic BH3 profiling 72 hours post treatment with perampanel or meclofenamate. FIG. 5C shows immunoblots of xenograft samples. FIG. 5D shows an immunoblot of xenograft tumors treated with perampanel. Quantification in graph is of the protein BIM.

FIG. 6A shows Ex-vivo BH3 profiling of orthotopic and subcutaneous lung cancer tumor, PC9. FIG. 6B & 6C show the fold change of tumor burden of orthotopic and subcutaneous tumors under Compound 1 treatment.

DETAILED DESCRIPTION OF THE INVENTION

In contrast to other tumors which can be therapeutically eradicated, GBM exist within the unique microenvironment of the central nervous system (CNS). Recently, a paradigmshifting discovery identified cancer cells connect with neurons in the brain microenvironment through neuron-glioma synapses to stimulate tumor growth (Lim-Fat and Wen, 2019). However, it is not known if neuro-glioma synapses drive therapeutic resistance, inevitably leading to the shortened survival of patients with brain tumors.

To investigate the intrinsic apoptotic machinery of GBM patient tumors take directly from their native environment, 29 unique patient glioma tumors were molecularly and functionally characterized by performing Whole Exome (WES) and RNA sequencing (RNA- seq) coupled with BH3 Profiling (FIG. 1A). Tumor specimens included purified patient tumor cells from both newly diagnosed and recurrent glioblastoma (GBM) (n=15) as well as lower grade gliomas (n=8). BH3 profiling and molecular analysis were performed on live, purified tumor cells following dissociation of freshly resected bulk tumor and removal of red blood cells (RBCs), myelin and CD45+ cells. The molecular analyses of both patient tumor cells and gliomaspheres samples captured the frequently altered GBM genetic alterations described by The Cancer Genome Atlas (TCGA) (Brennan et al., 2013), and represented the previously defined GBM subtypes (e.g., Proneural, Classical, Mesenchymal) (Verhaak et al., 2010) . A functional assessment of a tumor’s apoptotic potential (i.e., primed state) can strongly associate with therapy-induced intrinsic apoptosis (Certo et al., 2006b). The apoptotic potentials of the patient samples were then measured by preforming BH3 profiling with increasing concentrations of the pro-apoptotic BIM BH3 peptide (equal affinity to all anti-apoptotic proteins (Kale et al., 2018b)) (FIG. 1A). To determine if clinical features associate the primed state, patient samples were grouped into low grade (grades II and III) and high grade (grade IV). High grade tumors were less primed for apoptosis or more protected against cell death, relative to low grade tumors (FIG. IB). Clinically, low grade tumors have a better prognosis than high grade tumors (FIG. 1C). It was also assessed if tumor recurrence was associated with primed state by comparing the BH3 profiling of patient’s tumor at the 1st and 2nd recurrence. It was found that the tumor was less primed at the second recurrence (FIG. ID).

The impact of the tumor environment on the intrinsic apoptotic machinery was investigated by preforming cell BH3 profiling on tumors in the cell culture environment, orthotopic xenograft (mouse brain) and subcutaneous xenograft. First, when comparing patient samples to gliomaspheres in cell culture, it was observed that gliomaspheres were more primed for cell death (FIG. 2A). Gliomaspheres were used to create orthotopic xenografts and then ex- vivo BH3 profiling was performed. It was found that the orthotopic were more protected against cell death (FIG. 2B). To determine if this effect was brain specific, gliomaspheres were then used to create subcutaneous xenografts and again ex- vivo BH3 profiling was performed. It was found that similar to cell culture, tumors in the subcutaneous environment are more primed for cell death than tumors in the brain environment (FIG. 2C). To investigate if these differences in primed state correspond to drug sensitivity a fully brain penetrant EGFR inhibitor, Compound 1, which reaches equal concentrations in the brain and subcutaneous tumor tissue was selected (FIG. 2D). Next evaluated sensitivity to Compound 1 was evaluated by placing the same tumor, GBM39, into both the intracranial and subcutaneous environments and evaluating tumor burden. It was found that in the orthotopic environment, where the primed state was depressed, Compound 1 produced cytostatic response, slowing tumor growth relative to vehicle. In the subcutaneous environment, where the tumor is higher primed, Compound 1 exhibited a cytotoxic effect, shrinking tumor burden relative to when treatment began (FIG. 2E). To interrogate what the orthotopic xenograft environment was preserving that the cell culture and subcutaneous were not, differential gene expression was performed, and 40 genes were identified that were high in both patients and orthotopic xenografts and low in gliomaspheres and subcutaneous xenografts (FIG. 2F). Gene ontology revealed these genes were associated with neuron-glial interactions.

To determine if the neural-glioma synapses are directly responsible for promoting apoptotic resistance, neurons isolated were from pre-natal mice co-cultured. After 7 days gliomaspheres we added to the neurons and co-cultured for 72 hours (FIG. 3A) or gliomaspheres were cultured in conditioned media from the neurons for 72 hours. Gliomaspheres were then isolated from the neurons and used BH3 profiling. It was found that on direct neural co-culture and the conditioned media protects against apoptosis (FIG. 3B). Treatment with inhibitors that block synaptic signaling (e.g., TTX - inhibitor of voltage gates sodium channels; NBQX inhibits AMPA receptor) were able to significantly prime GBM cells for apoptosis. Next gliomaspheres co-cultured with neurons were treated with Meclofenamate, a gap junction blocker that prevents calcium waves from traveling between cells. Meclofenamate has a greater priming effect on gliomaspheres that are being co-cultured with neurons (FIG. 3D). Cell viability after cytotoxic drug treatment in the neuron co-culture environment was then evaluated (FIG. 3E). Baseline viability was no different between regular culture and neuron co-culture environments. After 7 days of treatment both Temozolomide (TMZ) and the EGFR inhibitor, Compound 1, reduced viability of gliomaspheres in regular cell culture conditions. Gliomaspheres in neuron co-culture conditions, treated with TMZ or EGFRi, had significantly increased viability. Next, it was assessed if the increased viability with the neuron co-cultured is a result a synaptic signaling. Co- treatment with Compound 1 with the synaptic signaling inhibitors, TTX or NBQX significantly reduced viable cells. Taken together, these data suggest that neural-GBM synapses promote apoptotic resistance in GBM.

Developing progenitor cells in the adult brain depend on synaptic signaling for survival, which converges on the transcription factor CREB. To begin to investigate if CREB activity is elevated in GBM tumors residing in the brain environment, tumors were scored for their expression of 64 CREB target genes, and it was found that tumors from the brain of the patient or from an orthotopic xenograft have stronger expression of CREB regulated genes than tumors in the cell culture environment (FIG. 4A). Then treated GBM cells being co-cultured with neurons with the CREB inhibitor, 666-15, and preformed BH3 profiling (FIG 4B). It was found that 666-15 decreased levels of CREB phosphorylation and primed the GBM tumor for cell death. It was then assessed how changes in the environment affect the BCL2 protein family, which together dictates the primed state of the tumor. Analysis of gene expression across environments revealed the anti-apoptotic protein, MCL1, loses expression in the cell culture environment (FIG. 4C). Then protein level of the BCL2 family was assessed in GBM tumors in traditional culture and neuron co-culture environments. Along with differences in MCL1 expression, we also observed BIM expression to deceased in GBM when cultured with neurons (FIG 4D). To test if this change is due to synaptic activity, the action potential blocker TTX was added to the GBM tumor and neuron co-culture. TTX treatment increased BIM expression in the GBM cells. (FIG 4E).

It was then investigated if the neural-glioma synapses are directly responsible for reducing GBM apoptotic potential in the brain microenvironment. Mice with orthotopic GBM xenografts were treated with perampanel for three days and then dynamic ex-vivo BH3 profiling was preformed to assess if the apoptotic potential shifts under treatment. It was found that the orthotopic tumors treated with perampanel demonstrated a stronger release of cytochrome c, indicating a higher apoptotic potential after their synaptic signaling was inhibited (FIG. 5A). These same experiments were then performed in the same model placed subcutaneously and it was found that perampanel did not alter the apoptotic potential outside the neural microenvironment (FIG. 5A). Dynamic ex-vivo BH3 profiling was then performed with the gap junction blocker, meclofenamate, and found that it also increased the primed state of an orthotopic xenograft (FIG 5B). Immunoblot analysis of these samples revealed that perampanel and meclofenamate treatment decreased the expression of the anti-apoptotic protein MCL-1 and increased expression of the pro-apoptotic activator BIM (FIG. 5C). It was also found that perampanel treatment only increased expression of BIM in the orthotopic xenograft, and not in the subcutaneous tumor (FIG 5D).

Lung cancers also form tumors in the brain environment. To investigate if these tumors are also afforded protection within the brain, cranial and subcutaneous xenografts were established with lung tumor cells, PC9, when ex-vivo BH3 profiling was performed on these tumors there was no difference in the basal primed state (FIG 6A). To test if these similar primed states result in similar therapeutic sensitivity, cranial and subcutaneous PC9 tumors were treated with Compound 1 and tumor burden was evaluated. In both the brain and subcutaneous environment, Compound 1 exhibited a cytotoxic effect, shrinking tumor burden relative to when treatment began (FIG. 6B). Together, these findings show that blocking neuroglioma synapses can alter the intrinsic apoptotic machinery to increase the apoptotic potential of GBM tumors.

In one aspect, the present disclosure provides methods treating brain cancer in a subject in need thereof, comprising administering an inhibitor of synaptic signaling to the subject. In certain embodiments, administering the inhibitor of synaptic signaling primes the brain cancer for apoptosis. In certain embodiments, the inhibitor of synaptic signaling is an inhibitor of voltage gated sodium channels, an inhibitor of the a-amino-3-hydroxy-5-methyl-4- isoxazolepropionic acid (AMP A) receptor, or a gap junction blocker.

In certain embodiments, the method further compirses administering an additional brain cancer therapy. In certain embodiments, the additional brain cancer therapy is selected from chemotherapy, tumor tearing fields, and radiation; or a combination thereof. In certian embodiments, the chemotherapy comprises a first EGFR inhibitor, a PI3K inhibitor, an mTOR inhibitor, a RAS inhibitor, a BRAF inhibitor, a HER2 inhibitor, a PDGFR inhibitor, a MET inhibitor, an ERK inhibitor, a MAPK inhibitor, avastin, irinotecan, temozolamide, procarbazine, carmustine, lomustine, or vincristine. In certain embodiments, the chemotherapy comprises a first EGFR inhibitor.

In aother aspect, the present disclosure provides methods of treating brain cancer in a subject in need thereof, comprising conjointly administering an inhibitor of synaptic signaling and a first EGFR inhibitor to the subject.

In certain embodiments, the first EGFR inhibitor is a compound of Formula I-a or

Formula I-b: or a pharmaceutically acceptable salt or stereoisomer thereof, wherein:

Z is aryl or heteroaryl;

R ¹ is hydrogen, alkyl, halo, CN, NO2, OR ⁷ , cycloalkyl, heterocyclyl, aryl or heteroaryl;

R ² is hydrogen, alkyl, halo, CN, NO2, OR ⁸ , cycloalkyl, heterocyclyl, aryl or heteroaryl; or R ¹ and R ² taken together complete a carbocyclic or heterocyclic ring;

R ³ is hydrogen, alkyl, or acyl;

R ⁴ is alkoxy;

R ⁵ is alkyl; and

R ⁷ and R ⁸ are each independently selected from hydrogen, alkyl, such as alkoxyalkyl, aralkyl, or arylacyl.

In certain embodiments, if R ⁷ and R ⁸ are alkoxyalkyl and R ³ is hydrogen, then Z is not 3-ethynylphenyl. In certain embodiments, Z is optionally substituted with R ⁶ selected from alkyl, alkoxy, OH, CN, NO2, halo, alkenyl, aralkyloxy, cycloalkyl, heterocyclyl, aryl, and heteroaryl.

In certain embodiments, either:

R ⁷ and R ⁸ are each independently selected from hydrogen, aralkyl, or arylacyl; each instance of R ⁶ is independently selected from alkyl, alkoxy, OH, CN, NO2, halo, alkenyl, aralkyloxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl; or

R ¹ and R ² taken together complete a carbocyclic or heterocyclic ring.

In certain embodiments, if R ⁷ and R ⁸ combine to form a heterocyclic ring and R ³ is hydrogen, then Z is not 2-fluoro-4-bromophenyl, 3-bromophenyl, 3-methylphenyl, 3- trifluoromethylphenyl, or 3-chloro-4-fluorophenyl.

In certain embodiments, R ¹ is hydrogen. In other embodiments, R ¹ is OR ⁷ .

In certain embodiments, R ⁷ is hydrogen. In other embodiments, R ⁷ is alkyl. In yet other embodiments, R ⁷ is alkoxyalkyl. In yet other embodiments, R ⁷ is arylacyl.

In certain embodiments, R ² is heteroaryl, such as furanyl. In certain embodiments, the heteroaryl of R ² is substituted with alkyl, alkoxy, OH, CN, NO2, halo, other embodiments, R ² is OR ⁸ .

In certain embodiments, R ⁸ is hydrogen. In other embodiments, R ⁸ is alkoxyalkyl. In other embodiments, R ⁸ is arylacyl.

In certain embodiments, R ¹ and R ² combine to form a carbocyclic or heterocyclic ring, such as a 5-member, 6-member, or 7-member carbocyclic or heterocyclic ring. In certain embodiments, the carbocyclic or heterocyclic ring resulting from the combination of R ¹ and R ² is substituted with hydroxyl, alkyl (e.g., methyl), or alkenyl (e.g., vinyl). In certain embodiments, the compound i

In certain embodiments, the carbocyclic or heterocyclic ring is substituted with alkyl

(e.g., methyl) and the alkyl moi eties are trans relative to each other.

In other embodiments, the compound

In certain embodiments, the carbocyclic or heterocyclic ring is substituted with alkyl

(e.g., methyl) and the alkyl moi eties are cis relative to each other.

In yet other embodiments, the compound

In certain embodiments, the compound is a compound of Formula (Ill-a), (Ill-b), (III- c), (Ill-d), (Ill-e), or (III-f):

In certain embodiments, R ³ is hydrogen. In other embodiments, R ³ is acyl. In certain embodiments, R ³ is alkylacyl. In certain embodiments, R ³ is alkoxyacyl. In certain embodiments, R ³ is acyloxyalkyl. In certain embodiments, alkyl.

In certain embodiments, Z is aryl or heteroaryl optionally substituted with one or more R ⁶ ; and each instance of R ⁶ is independently selected from alkyl, alkoxy, OH, CN, NO2, halo, alkenyl, alkynyl, aralkyloxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl. In certain embodiments, Z is phenyl substituted with 1, 2, 3, 4, or 5 R ⁶ . In certain embodiments, each R ⁶ is independently selected from halo, alkyl, alkynyl, or arylalkoxy. In certain embodiments, Z is 2-fluoro-3-chlorophenyl, 2-fluorophenyl, 2,3 -difluorophenyl, 2,4-difluorophenyl, 2,5- difluorophenyl, 2,6-difluorophenyl, 2,4,6-trifluorophenyl, pentafluorophenyl, 2-fluoro-3- bromophenyl, 2-fluoro-3-ethynylphenyl, and 2-fluoro-3-(trifluoromethyl)phenyl. In certain preferred embodiments, 2-fluoro-3-bromophenyl. In other embodiments, Z is 3-ethynylphenyl. In yet other embodiments, Z is 3-chloro-4-((3-fluorobenzyl)oxy)benzene. In yet other embodiments, 3-chloro-2-(trifluoromethyl)phenyl. In other embodiments, Z is 2-fluoro-5- bromophenyl. In yet other embodiments, Z is 2,6-difluoro-5-bromophenyl. In certain embodiments, Z is substituted with one R ⁶ selected from and R ¹⁰ are each independently alkyl.

In certain embodiments, the compound is a compound of Formula (IV-a):

and each R ⁶ is independently selected from fluoro, chloro, or bromo.

In certain embodiments, the compound is a compound of Formula (IV-b): and each R ⁶ is independently selected from fluoro, chloro, or bromo.

In certain embodiments, the compound is a compound of Formula (IV-c): and each R ⁶ is independently selected from fluoro, chloro, or bromo.

In certain embodiments, the compound is a compound of Formula (IV-a): and each R ⁶ is independently selected from fluoro, chloro, or bromo.

In certain embodiments, the compound is a compound of Formula (V-b): and each R ⁶ is independently selected from fluoro, chloro, or bromo.

In certain embodiments, the compound is a compound of Formula (V-c): and each R ⁶ is independently selected from fluoro, chloro, or bromo.

In certain embodiments, the first EGFR inhibitor is: -91 - pharmaceutically acceptable salt or stereoisomer thereof.In certain embodiments, the first EGFR inhibitor is a compound of Formula VI or Formula VI*: or a pharmaceutically acceptable salt thereof, wherein:

Z is aryl or heteroaryl;

R ^2a and R ^2b are each independently selected from hydrogen, alkyl, halo, CN, and NO2;

R ³ is hydrogen, alkyl, or acyl;

R ⁴ is alkoxy;

R ⁵ is alkyl; R ⁷ and R ⁸ are, each independently, selected from hydrogen, alkyl, such as alkoxyalkyl, aralkyl, or arylacyl;

R ¹¹ is hydrogen, alkyl, halo, CN, NO2, OR ⁷ , cycloalkyl, heterocyclyl, aryl or heteroaryl; and R ¹² is hydrogen, alkyl, halo, CN, NO2, OR ⁸ , cycloalkyl, heterocyclyl, aryl or heteroaryl; or

R ¹¹ and R ¹² taken together complete a carbocyclic or heterocyclic ring.

In certain embodiments, if R ^2a is hydrogen, then R ^2b is selected from alkyl, halo, CN, and NO2. In certain embodiments, if R ^2b is hydrogen, then R ^2a is selected from alkyl, halo, CN, and NO2.

In certain embodiments, the compound is a compound of Formula (Vila) or Formula

(Vllb): (Vila) (Vllb) or a pharmaceutically acceptable salt thereof, wherein each instance of R ⁶ is independently selected from alkyl, alkoxy, OH, CN, NO2, halo, alkenyl, alkynyl, aralkyloxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

In certain embodiments, R ¹¹ is hydrogen. In other embodiments, R ¹¹ is OR ⁷ . In certain embodiments, R ⁷ is hydrogen. In other embodiments, R ⁷ is alkyl. In yet other embodiments, R ⁷ is alkoxyalkyl. In yet other embodiments, R ⁷ is arylacyl.

In certain embodiments, R ¹² is heteroaryl, such as furanyl. In certain embodiments, the heteroaryl of R ¹² is substituted with alkyl, alkoxy, OH, CN, NO2, halo, other embodiments, R ¹² is OR ⁸ . In certain embodiments, R ⁸ is hydrogen. In other embodiments, R ⁸ is alkoxyalkyl. In certain embodiments, R ⁸ is alkyl substituted certain embodiments, R ⁸ is acyl. In certain embodiments, R ¹¹ and R ¹² combine to form a carbocyclic or heterocyclic ring, such as a 5-member, 6-member, or 7-member carbocyclic or heterocyclic ring. In certain embodiments, the carbocyclic or heterocyclic ring arising from the combination of R ¹¹ and R ¹² is substituted with hydroxyl, alkyl (e.g., methyl), or alkenyl (e.g., vinyl).

In certain preferred embodiments, the first EGFR inhibitor is a compound of Formula Via, VIb, Vic, or Vid:

or a pharmaceutically acceptable salt thereof, wherein:

X is O, S, orNH;

Z is aryl or heteroaryl;

R ¹ is hydrogen or alkyl;

R ^2a and R ^2b are each independently selected from hydrogen, alkyl, halo, CN, and NO2;

R ³ is hydrogen, alkyl, or acyl;

R ⁴ is alkoxy;

R ⁵ is alkyl; and n is 0-3.

In certain embodiments, either R ^2a or R ^2b is selected from alkyl, halo, CN, and NO2.

In certain embodiments, the compound is a compound of Formula (Villa) or Formula (Vlllb):

(Vllla) (Vlllb) or a pharmaceutically acceptable salt thereof, wherein each instance of R ⁶ is independently selected from alkyl, alkoxy, OH, CN, NO2, halo, alkenyl, alkynyl, aralkyloxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

In certain preferred embodiments, R ¹ is alkyl (e.g., methyl or ethyl) substituted with heterocyclyl (e.g., a nitrogen-containing heterocyclyl, such as morpholinyl, piperidinyl, pyrrolodinyl, or piperazinyl, such as N-methyl piperazinyl). In certain other embodiments, R ¹ is alkyl (e.g., methyl or ethyl) substituted with amino (e.g., dimethyl amino). In certain further preferred embodiments, R ¹ is represented by Formula IX: wherein,

R ^13a and R ^13b are each independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; or R ^13a and R ^13b combine to form a heterocyclyl; and y is 0-3.

In certain embodiments, R ¹ is alkyl (e.g., methyl or ethyl) substituted with hydroxyl.

In certain embodiments, R ¹ is in the S configuration. In other embodiments, R ¹ is in the R configuration.

In certain embodiments, R ³ is hydrogen. In other embodiments, R ³ is acyl. In yet other embodiments, R ³ is alkylacyl. In yet other embodiments, R ³ is alkyloxyacyl. In yet other embodiments, R ³ is acyloxyalkyl. In yet other embodiments, alkyl.

In certain embodiments, Z is 2-fluoro-3-chlorophenyl, 2-fluorophenyl, 2,3- difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 2,4,6- trifluorophenyl, pentafluorophenyl, 2-fluoro-3-bromophenyl, 2-fluoro-3-ethynylphenyl, and 2-fluoro-3-(trifluoromethyl)phenyl. In certain embodiments, Z is 3-ethynylphenyl. In certain embodiments, Z is 3-chloro-4-((3-fluorobenzyl)oxy)benzene. In certain embodiments, Z is 3- chloro-2-(trifluoromethyl)phenyl. In certain preferred embodiments, Z is 2-fluoro-3- bromophenyl. In certain embodiments, Z is 2-fluoro-5-bromophenyl. In certain embodiments, Z is 2,6-difluoro-5-bromophenyl. In certain embodiments, Z is substituted with one R ⁶ selected from

R ⁹ and R ¹⁰ are each independently alkyl.

In certain embodiments, the compound is a compound of Formula (Xa): or a pharmaceutically acceptable salt thereof, wherein each R ⁶ is independently selected from fluoro, chloro, or bromo.

In certain embodiments, the compound is a compound of Formula (Xb): or a pharmaceutically acceptable salt thereof, wherein each R ⁶ is independently selected from fluoro, chloro, or bromo.

In certain embodiments, the compound is a compound of Formula (Xc): or a pharmaceutically acceptable salt thereof, wherein each R ⁶ is independently selected from fluoro, chloro, or bromo.

In certain embodiments, R ^2a is hydrogen. In certain embodiments, R ^2a is halo (e.g., fluoro).

In certain embodiments, R ^2b is hydrogen. In certain embodiments, R ^2b is halo (e.g., fluoro).

In certain embodiments, the first EGFR inhibitor is: pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is:

pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is: a pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is: a pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is: a pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor i pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor i a pharmaceutically acceptable salt thereof. In certain embodiments, the first EGFR inhibitor pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor i or a pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is: a pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor a pharmaceutically acceptable salt thereof. In certain embodiments, the first EGFR inhibitor pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor i or a pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor i pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor pharmaceutically acceptable salt thereof. In certain embodiments, the first EGFR inhibitor i a pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor a pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is a compound of Formula (XI): wherein: R ¹ is selected from

R ² is selected from Ci-Ce alkyl and Cs-Ce cycloalkyl, each of which is optionally substituted with one or more halogen, or a pharmaceutically acceptable salt thereof.

In certain embodiments, pharmaceutically acceptable salt thereof. In other embodiments, pharmaceutically acceptable salt thereof. In yet other embodiments, pharmaceutically acceptable salt thereof. In yet other embodiments, pharmaceutically acceptable salt thereof. In yet other embodiments, pharmaceutically acceptable salt thereof. In yet other embodiments, pharmaceutically acceptable salt thereof.

In certain embodiments, R ² is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, hexyl, trifluoromethyl, fluoroethyl, and difluoroethyl. In certain preferred embodiments, R ² is selected from methyl, ethyl, n-propyl, isopropyl, tert-butyl, fluoroethyl, and difluoroethyl. In even further preferred embodiments, R ² is methyl.

In certain embodiments, the first EGFR inhibitor is a compound of Formula (Xia):

R ² is selected from Ci-Ce alkyl and Cs-Ce cycloalkyl, each of which is optionally substituted with one or more halogen, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the first EGFR inhibitor is a compound with a structure represented by Formula (Xlb):

R ² is selected from Ci-Ce alkyl and Cs-Ce cycloalkyl, each of which is optionally substituted with one or more halogen, or a pharmaceutically acceptable salt thereof.

In certain preferred embodiments, the first EGFR inhibitor is:

Compound 1.

In certain embodiments, the additional brain cancer therapy is radiation.

In certain embodiments, the inhibitor of synaptic signaling is an inhibitor of voltage gated sodium channels. In certain embodiments, the inhibitor of voltage gated sodium channels is an inhibitor of NaVl.l, NaV1.2, NaV1.3, NaV1.4, NaV1.5, NaV1.6, NaV.17, NaV1.8, NaV1.9, or NaX. In certain embodiments, the inhibitor of voltage gated sodium channels is carbamazepine, phenytoin, mexiletine, flecainide, lidocaine, bupivacaine, ralfmamide, lacosamide, or tetrodotoxin. In certain embodiments, the inhibitor of voltage gated sodium channels is tetrodotoxin.

In certain embodiments, the inhibitor of synaptic signaling is an inhibitor of AMP A. In certain embodiments, the inhibitor of AMPA is CNQX, CP 465022, DNQX, GYKI 52466, GYKI 53655, IEM 1925, naspm trihydrochloride, NBQX, philanthotoxin 74, talampanel, ZK 200775m aniracetam, IEM, 1460, or perampanel. In certain embodiments, the inhibitor of AMPA is perampanel or NBQX.

In certain embodiments, the inhibitor of synaptic signaling is a gap junction blocker. In certain embodiments, the gap junction blocker is carbenoxolone, quinine, mefloquine, quinidine, anandamide, oleamide, meclofenamic acid, niflumic acid, flufenamic acid, heptanol, glycyrrhetinic acid, retinoic acid, or meclofenamate. In certain embodiment, the gap junction blocker is meclofenamate.

In certain embodiments, the brain cancer is glioblastoma, glioblastoma multiforme, glioma, low-grade astrocytoma, mixed oligoastrocytoma, pilocytic astrocytoma, pleomorphic xanthoastrocytoma, subependymal giant cell astrocytoma, or anaplastic astrocytoma. In certain preferred embodiments, the brain cancer is glioblastoma. In certain embodiments, the brain cancer is a high-grade glioblastoma. In other embodiments, the brain cancer is a low- grade glioblastoma.

In certain embodiments, the brain cancer is EGFR amplified. In certain embodiments, the brain cancer is positive for NF1 WT. In certain embodiments, the brain cancer is positive for aNFl mutant. In certain embodiments, the brain cancer is positive for PTEN WT. In certain embodiments, the brain cancer is positive for a PTEN mutant. In certain embodiments, the brain cancer is negative for PTEN.

In certain embodiments, the brain cancer is relapsed. In certain embodiments, the brain cancer is refractory.

In certain embodiments, the brain cancer is refractory to an EGFR inhibitor (e.g., osimertinib, erlotinib, afatinib, gefitinib, or dacomitinib). In certain embodiments, the brain cancer is refractory to one or more of osimertinib, erlotinib, afatinib, gefitinib, or dacomitinib. In certain embodiments, the brain cancer is refractory to osimertinib.

In certain embodiments, the subject has previously received a treatment with a second EGFR inhibitor (e.g., osimertinib, erlotinib, afatinib, gefitinib, or dacomitinib). In certain embodiments, the previous treatment with the second EGFR inhibitor (e.g., osimertinib, erlotinib, afatinib, gefitinib, or dacomitinib) failed.

In certain embodiments, the second EGFR inhibitor is osimertinib, erlotinib, afatinib, gefitinib, or dacomitinib. In certain embodiments, the second EGFR inhibitor is osimertinib. In certain embodiments, the second EGFR inhibitor is an EGFR inhibitor defined herein. In certain embodiments, the second EGFR inhibitor is the EGFR inhibitor is Compound 1.

Pharmaceutical Compositions

The compositions and methods of the present invention may be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or the compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are for human administration, particularly for invasive routes of administration (i.e., routes, such as injection or implantation, that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipients can be chosen, for example, to effect delayed release of an agent or to selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form such as tablet, capsule (including sprinkle capsule and gelatin capsule), granule, lyophile for reconstitution, powder, solution, syrup, suppository, injection or the like. The composition can also be present in a transdermal delivery system, e.g., a skin patch. The composition can also be present in a solution suitable for topical administration, such as a lotion, cream, or ointment.

A pharmaceutically acceptable carrier can contain physiologically acceptable agents that act, for example, to stabilize, increase solubility or to increase the absorption of a compound such as a compound of the invention. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a selfemulsifying drug delivery system or a selfmicroemulsifying drug delivery system. The pharmaceutical composition (preparation) also can be a liposome or other polymer matrix, which can have incorporated therein, for example, a compound of the invention. Liposomes, for example, which comprise phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.

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

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

A pharmaceutical composition (preparation) can be administered to a subject by any of a number of routes of administration including, for example, orally (for example, drenches as in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (for example as a patch applied to the skin); and topically (for example, as a cream, ointment or spray applied to the skin). The compound may also be formulated for inhalation. In certain embodiments, a compound may be simply dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable for same can be found in, for example, U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, as well as in patents cited therein.

The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about ninety-nine percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

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

Formulations of the invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), lyophile, powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. Compositions or compounds may also be administered as a bolus, electuary or paste.

To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as, modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surfaceactive or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butylene glycol, oils (in particular, cottonseed, groundnut, com, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

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

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

Dosage forms for the topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required.

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

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

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

The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrastemal injection and infusion. Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

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

These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

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

For use in the methods of this invention, active compounds can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.

Methods of introduction may also be provided by rechargeable or biodegradable devices. Various slow release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form an implant for the sustained release of a compound at a particular target site.

Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound(s) being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the pharmaceutical composition or compound at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. By “therapeutically effective amount” is meant the concentration of a compound that is sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the weight, sex, age, and medical history of the subject. Other factors which influence the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound of the invention. A larger total dose can be delivered by multiple administrations of the agent. Methods to determine efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13 ed., 1814-1882, herein incorporated by reference).

In general, a suitable daily dose of an active compound used in the compositions and methods of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In certain embodiments of the present invention, the active compound may be administered two or three times daily. In preferred embodiments, the active compound will be administered once daily.

The patient receiving this treatment is any animal in need, including primates, in particular humans; and other mammals such as equines, cattle, swine, sheep, cats, and dogs; poultry; and pets in general. In certain embodiments, compounds of the invention may be used alone or conjointly administered with another type of therapeutic agent.

The present disclosure includes the use of pharmaceutically acceptable salts of compounds of the invention in the compositions and methods of the present invention. In certain embodiments, contemplated salts of the invention include, but are not limited to, alkyl, dialkyl, trialkyl or tetra-alkyl ammonium salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, L-arginine, benenthamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2- (diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, IH-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, l-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, 1 -hydroxy -2 -naphthoic acid, 2, 2-di chloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, 1-ascorbic acid, 1-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, 1-malic acid, malonic acid, mandelic acid, methanesulfonic acid , naphthalene-l,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, proprionic acid, 1- pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, 1-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid acid salts.

The pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent. Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Definitions

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

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

Chemistry terms used herein, unless otherwise defined herein, are used according to conventional usage in the art, as exemplified by “The McGraw-Hill Dictionary of Chemical Terms”, Parker S., Ed., McGraw-Hill, San Francisco, C.A. (1985).

All of the above, and any other publications, patents and published patent applications referred to in this application are specifically incorporated by reference herein. In case of conflict, the present specification, including its specific definitions, will control. The term “agent” is used herein to denote a chemical compound (such as an organic or inorganic compound, a mixture of chemical compounds), a biological macromolecule (such as a nucleic acid, an antibody, including parts thereof as well as humanized, chimeric and human antibodies and monoclonal antibodies, a protein or portion thereof, e.g., a peptide, a lipid, a carbohydrate), or an extract made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues. Agents include, for example, agents whose structure is known, and those whose structure is not known.

A “patient,” “subject,” or “individual” are used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (including bovines, porcines, etc.), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats).

“Treating” a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.

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

“Administering” or “administration of’ a substance, a compound or an agent to a subject can be carried out using one of a variety of methods known to those skilled in the art. For example, a compound or an agent can be administered, intravenously, arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (by absorption, e.g., through a skin duct). A compound or agent can also appropriately be introduced by rechargeable or biodegradable polymeric devices or other devices, e.g., patches and pumps, or formulations, which provide for the extended, slow or controlled release of the compound or agent. Administering can also be performed, for example, once, a plurality of times, and/or over one or more extended periods.

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

As used herein, the phrase “conjoint administration” refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered therapeutic agent is still effective in the body (e.g., the two agents are simultaneously effective in the patient, which may include synergistic effects of the two agents). For example, the different therapeutic compounds can be administered either in the same formulation or in separate formulations, either concomitantly or sequentially. Thus, an individual who receives such treatment can benefit from a combined effect of different therapeutic agents.

A “therapeutically effective amount” or a “therapeutically effective dose” of a drug or agent is an amount of a drug or an agent that, when administered to a subject will have the intended therapeutic effect. The full therapeutic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount needed for a subject will depend upon, for example, the subject’s size, health and age, and the nature and extent of the condition being treated, such as cancer or MDS. The skilled worker can readily determine the effective amount for a given situation by routine experimentation.

As used herein, the terms “optional” or “optionally” mean that the subsequently described event or circumstance may occur or may not occur, and that the description includes instances where the event or circumstance occurs as well as instances in which it does not. For example, “optionally substituted alkyl” refers to the alkyl may be substituted as well as where the alkyl is not substituted.

It is understood that substituents and substitution patterns on the compounds of the present invention can be selected by one of skill in the art to result chemically stable compounds which can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results.

As used herein, the term “optionally substituted” refers to the replacement of one to six hydrogen radicals in a given structure with the radical of a specified substituent including, but not limited to: hydroxyl, hydroxyalkyl, alkoxy, halogen, alkyl, nitro, silyl, acyl, acyloxy, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OCO-CH2-O- alkyl, -OP(O)(O-alkyl)2 or -CH2-OP(O)(O-alkyl)2. Preferably, “optionally substituted” refers to the replacement of one to four hydrogen radicals in a given structure with the substituents mentioned above. More preferably, one to three hydrogen radicals are replaced by the substituents as mentioned above. It is understood that the substituent can be further substituted.

As used herein, the term “alkyl” refers to saturated aliphatic groups, including but not limited to C1-C10 straight-chain alkyl groups or C1-C10 branched-chain alkyl groups. Preferably, the “alkyl” group refers to Ci-Ce straight-chain alkyl groups or Ci-Ce branched-chain alkyl groups. Most preferably, the “alkyl” group refers to C1-C4 straight-chain alkyl groups or C1-C4 branched-chain alkyl groups. Examples of “alkyl” include, but are not limited to, methyl, ethyl, 1 -propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, 1 -pentyl, 2-pentyl, 3-pentyl, neo-pentyl, 1- hexyl, 2-hexyl, 3-hexyl, 1 -heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1 -octyl, 2-octyl, 3-octyl or 4- octyl and the like. The “alkyl” group may be optionally substituted.

The term “acyl” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

The term “acylamino” is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbylC(O)NH-.

The term “acyloxy” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O-, preferably alkylC(O)O-.

The term “alkoxy” refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like. The term “alkoxyalkyl” refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

The term “alkyl” refers to saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In preferred embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., Ci- 30 for straight chains, C3-30 for branched chains), and more preferably 20 or fewer.

Moreover, the term “alkyl” as used throughout the specification, examples, and claims is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2- trifluoroethyl, etc.

The term “C _x-y ” or “C _x -C _y ”, when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups that contain from x to y carbons in the chain. Coalkyl indicates a hydrogen where the group is in a terminal position, a bond if internal. A Ci-ealkyl group, for example, contains from one to six carbon atoms in the chain.

The term “alkylamino”, as used herein, refers to an amino group substituted with at least one alkyl group.

The term “alkylthio”, as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS-.

The term “amido”, as used herein, refers to a group wherein R ⁹ and R ¹⁰ each independently represent a hydrogen or hydrocarbyl group, or R ⁹ and R ¹⁰ taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

The terms “amine” and “amino” are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by wherein R ⁹ , R ¹⁰ , and R ¹⁰ ’ each independently represent a hydrogen or a hydrocarbyl group, or R ⁹ and R ¹⁰ taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

The term “aminoalkyl”, as used herein, refers to an alkyl group substituted with an amino group.

The term “aralkyl”, as used herein, refers to an alkyl group substituted with an aryl group.

The term “aryl” as used herein include substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon. Preferably the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term “aryl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

The term “carbamate” is art-recognized and refers to a group wherein R ⁹ and R ¹⁰ independently represent hydrogen or a hydrocarbyl group.

The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

The term “carbocycle” includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated and aromatic rings. Carbocycle includes bicyclic molecules in which one, two or three or more atoms are shared between the two rings. The term “fused carbocycle” refers to a bicyclic carbocycle in which each of the rings shares two adjacent atoms with the other ring. Each ring of a fused carbocycle may be selected from saturated, unsaturated and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, is included in the definition of carbocyclic. Exemplary “carbocycles” include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct- 3-ene, naphthalene and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-lH- indene and bicyclo[4.1.0]hept-3-ene. “Carbocycles” may be substituted at any one or more positions capable of bearing a hydrogen atom.

The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

The term “carbonate” is art-recognized and refers to a group -OCO2-.

The term “carboxy”, as used herein, refers to a group represented by the formula -CO2H.

The term “cycloalkyl” includes substituted or unsubstituted non-aromatic single ring structures, preferably 4- to 8-membered rings, more preferably 4- to 6-membered rings. The term “cycloalkyl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is cycloalkyl and the substituent (e.g., R ¹⁰⁰ ) is attached to the cycloalkyl ring, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, denzodioxane, tetrahydroquinoline, and the like.

The term “ester”, as used herein, refers to a group -C(O)OR ⁹ wherein R ⁹ represents a hydrocarbyl group.

The term “ether”, as used herein, refers to a hydrocarbyl group linked through an oxygen to another hydrocarbyl group. Accordingly, an ether substituent of a hydrocarbyl group may be hydrocarbyl-O-. Ethers may be either symmetrical or unsymmetrical. Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include “alkoxyalkyl” groups, which may be represented by the general formula alkyl-O-alkyl.

The terms “halo” and “halogen” as used herein means halogen and includes chloro, fluoro, bromo, and iodo.

The terms “hetaralkyl” and “heteroaralkyl”, as used herein, refers to an alkyl group substituted with a hetaryl group.

The terms “heteroaryl” and “hetaryl” include substituted or unsubstituted aromatic single ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heteroaryl” and “hetaryl” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.

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

The term “heterocyclylalkyl”, as used herein, refers to an alkyl group substituted with a heterocycle group.

The terms “heterocyclyl”, “heterocycle”, and “heterocyclic” refer to substituted or unsubstituted non-aromatic ring structures, preferably 3- to 10-membered rings, more preferably 3- to 7-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heterocyclyl” and “heterocyclic” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

The term “hydrocarbyl”, as used herein, refers to a group that is bonded through a carbon atom that does not have a =0 or =S substituent, and typically has at least one carbonhydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Thus, groups like methyl, ethoxyethyl, 2-pyridyl, and even trifluoromethyl are considered to be hydrocarbyl for the purposes of this application, but substituents such as acetyl (which has a =0 substituent on the linking carbon) and ethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbyl groups include, but are not limited to aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

The term “hydroxyalkyl”, as used herein, refers to an alkyl group substituted with a hydroxy group.

The term “lower” when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups where there are ten or fewer atoms in the substituent, preferably six or fewer. A “lower alkyl”, for example, refers to an alkyl group that contains ten or fewer carbon atoms, preferably six or fewer. In certain embodiments, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are respectively lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, such as in the recitations hydroxyalkyl and aralkyl (in which case, for example, the atoms within the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

The terms “polycyclyl”, “poly cycle”, and “polycyclic” refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls) in which two or more atoms are common to two adjoining rings, e.g., the rings are “fused rings”. Each of the rings of the poly cycle can be substituted or unsubstituted. In certain embodiments, each ring of the poly cycle contains from 3 to 10 atoms in the ring, preferably from 5 to 7.

The term “sulfate” is art-recognized and refers to the group -OSOsH, or a pharmaceutically acceptable salt thereof.

The term “sulfonamido” is art-recognized and refers to the group represented by the general formulae wherein R ⁹ and R ¹⁰ independently represents hydrogen or hydrocarbyl.

The term “sulfoxide” is art-recognized and refers to the group-S(O)-.

The term “sulfonate” is art-recognized and refers to the group SOsH. or a pharmaceutically acceptable salt thereof.

The term “sulfone” is art-recognized and refers to the group -S(O)2-.

The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.

The term “thioalkyl”, as used herein, refers to an alkyl group substituted with a thiol group.

The term “thioester”, as used herein, refers to a group -C(O)SR ⁹ or -SC(O)R ⁹ wherein R ⁹ represents a hydrocarbyl.

The term “thioether”, as used herein, is equivalent to an ether, wherein the oxygen is replaced with a sulfur.

The term “urea” is art-recognized and may be represented by the general formula wherein R ⁹ and R ¹⁰ independently represent hydrogen or a hydrocarbyl.

The term “modulate” as used herein includes the inhibition or suppression of a function or activity (such as cell proliferation) as well as the enhancement of a function or activity.

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

“Pharmaceutically acceptable salt” or “salt” is used herein to refer to an acid addition salt or a basic addition salt which is suitable for or compatible with the treatment of patients.

The term “pharmaceutically acceptable acid addition salt” as used herein means any non-toxic organic or inorganic salt of any base compounds represented by Formula I. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric and phosphoric acids, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, benzoic, phenylacetic, cinnamic and salicylic acids, as well as sulfonic acids such as p-toluene sulfonic and methanesulfonic acids. Either the mono or di-acid salts can be formed, and such salts may exist in either a hydrated, solvated or substantially anhydrous form. In general, the acid addition salts of compounds of Formula I are more soluble in water and various hydrophilic organic solvents, and generally demonstrate higher melting points in comparison to their free base forms. The selection of the appropriate salt will be known to one skilled in the art. Other non- pharmaceutically acceptable salts, e.g., oxalates, may be used, for example, in the isolation of compounds of Formula I for laboratory use, or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

The term “pharmaceutically acceptable basic addition salt” as used herein means any non-toxic organic or inorganic base addition salt of any acid compounds represented by Formula I or any of their intermediates. Illustrative inorganic bases which form suitable salts include lithium, sodium, potassium, calcium, magnesium, or barium hydroxide. Illustrative organic bases which form suitable salts include aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine and picoline or ammonia. The selection of the appropriate salt will be known to a person skilled in the art.

Many of the compounds useful in the methods and compositions of this disclosure have at least one stereogenic center in their structure. This stereogenic center may be present in a R or a S configuration, said R and S notation is used in correspondence with the rules described in Pure Appl. Chem. (1976), 45, 11-30. The disclosure contemplates all stereoisomeric forms such as enantiomeric and diastereoisomeric forms of the compounds, salts, prodrugs or mixtures thereof (including all possible mixtures of stereoisomers). See, e.g., WO 01/062726.

Furthermore, certain compounds which contain alkenyl groups may exist as Z (zusammen) or E (entgegen) isomers. In each instance, the disclosure includes both mixture and separate individual isomers.

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

The phrase “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filter, diluent, excipient, solvent or encapsulating material useful for formulating a drug for medicinal or therapeutic use.

The term “Log of solubility”, “LogS” or “logS” as used herein is used in the art to quantify the aqueous solubility of a compound. The aqueous solubility of a compound significantly affects its absorption and distribution characteristics. A low solubility often goes along with a poor absorption. LogS value is a unit stripped logarithm (base 10) of the solubility measured in mol/liter.

The phrase “high grade glioblastoma” as used herein means a grade 3 or grade 4 glioblastoma.

The phrase “low grade glioblastoma” as used herein means a grade 1 or grade 2 glioblastoma.

EXAMPLES

The invention now being generally described, it will be more readily understood by reference to the following examples which are included merely for purposes of illustration of certain aspects and embodiments of the present invention and are not intended to limit the invention.

Example 1: Exemplary Treatment of NSCLC Xenograft with Compound 1

Secreted gaussia luciferase expressing PC9 cells, which were derived from EGFR exon 19 deletion NSCLC, were intracranially injected into female NSG mice and were randomized into the following treatment groups: vehicle, osimertinib at 10 and 25 mg/kg (mpk) and Compound 1 at 10 and 25 mpk (n=8 mice per group). Dosing schedule was QD for 5 days followed by 2 days off. Relative light unit (RLU) intensity from tumor secreted gaussia luciferase was measured as a surrogate of intracranial tumor growth. RLU and body weight were measured 2x weekly until mice were taken down due to body weight loss, health observations, and/or study termination.

As of treatment day 91, Compound 1 at 10 and 25 mpk both showed an extension of survival relative to vehicle of >450%. Osimertinib at 10 and 25 mpk showed an extension of survival of 264% and >450% relative to vehicle, respectively. The median survival for vehicle treatment was 16.5 days. Median survival for either Compound 1 dose and osimertinib at 25 mpk has not been reached and the median survival for osimertinib at 10 mpk is 60 days. At either dose, Compound 1 achieved significantly greater tumor growth inhibition than osimertinib at 25 mpk (p-value < 0.05). Osimertinib at 10 and 25 mpk achieved a maximum tumor regression of 37% and 75% on treatment day 17, respectively. On this treatment day, Compound 1 at 10 and 25 mpk achieved a significantly greater tumor regression of 89% and 94% relative to osimertinib at 25 mpk (p-values < 0.05). At 25 mpk in a mouse PK study, Compound 1 achieved an unbound brain peak concentration (Cmax) of 400 nM and osimertinib achieved 20 nM. In a 3-day cellular viability assay in the PC9 cell line, Compound 1 had an IC50 of 33 nM while osimertinib had an IC50 of 31 nM.

Compound 1 at 10 and 25 mpk achieved significantly greater tumor regressions than osimertinib at its MTD mouse dose of 25 mpk in an intracranial EGFR exon 19 deletion NSCLC PC9 CDX study. Exhibiting both potent activity against EGFR alterations and high CNS penetration, Compound 1 shows promising nonclinical activity in cancers outside of GBM, such as CNS metastases of EGFR mutant NSCLC.

Table 1. Comparison of the brain penetration of Erlotinib. Osimertinib. and Compound 1 Table 2, Comparison of the pharmacokinetic properties of Osimertinib. and Compound 1

INCORPORATION BY REFERENCE

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

EQUIVALENTS

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