Use of an ilmotepride compound in the manufacture of a medicament for the treatment of a central nervous system cancer

Abstract

The present application relates to the use of an imremeritide compound of Formula (1) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a central nervous system cancer in a subject, wherein the central nervous system cancer has a histone H3 mutation.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 201780015327.8, filed January 30, 2017, entitled "MODULATION OF G PROTEIN-COUPLED RECEPTORS (GPCRs) BY IMIPRIDONES". TECHNICAL FIELD

[0002] The present application relates to the use of an imipridone compound or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a central nervous system cancer in a subject, wherein the central nervous system cancer has a histone H3 mutation. BACKGROUND

[0003] Human cells have various receptors on their surfaces. G protein-coupled receptors ("GPCRs") form one of the largest families of transmembrane receptor proteins. The human genome has about 30,000 genes, of which up to 1,000 encode GPCRs. GPCRs are divided into five classes. The first class is the rhodopsin receptor family or "Class A GPCRs" of 670 receptor proteins. The rhodopsin receptor family can react with a variety of ligands including amines (alpha group), peptides (beta group), lipid-like substances (gamma group), nucleotides and glycoproteins (delta group), and contains many drug target receptors. The second class is the secretin receptor family and has binding domains for peptide hormones. The receptors in this family are involved in homeostasis in the body and have become important targets for drug development. The third class is the adhesion receptor family, characterized by a GPCR proteolytic site (GPS). Drug development targeting this GPCR family has not occurred because they exhibit a variety of N-terminal portions and little is known about their ligands. The fourth class is the glutamate receptor family, of which 22 GPCR members have been identified to date. Little is known about the specificity of each protein. The last class is the Frizzled / Taste2 family, which includes 10 Frizzled receptors that use Wnt glycoproteins as ligands, 5 SMO (smoothened) receptors that do not require a ligand, and 25 Taste2 receptors that are required to perceive various tastes. Receptors including GPCRs are also classified based on the identification of endogenous ligands. Receptors bind to known endogenous compounds or are classified as orphan receptors for which endogenous ligands have not been identified.

[0004] GPCRs are present in a wide range of tissues and cell types and are associated with many different physiological mechanisms. They are activated by a wide variety of ligands, such as hormones such as thyroid stimulating hormone (TSH), adrenocorticotropic hormone, glucagon, and vasopressin, amines such as 5-HT, acetylcholine (muscarinic AchR), and histamine, lipids such as LPA and S1P, and signal transmitters such as amino acids, Ca2 +GPCRs, nucleic acids, peptides and light. The wide distribution and diversity of the roles played by GPCRs demonstrate their importance in a variety of pathological conditions. Indeed, GPCRs are involved in a variety of diseases, including bronchoconstriction, hypertension, diabetes, inflammation, cell death, hormonal disorders, cancer, neurotransmission and behavioral disorders. GPCRs are therefore an important area for drug development. It is currently believed that approximately 360 GPCRs are available for drug development. Of these, 46 have already been used for drug development. There are an estimated 150 orphan GPCRs (oGPCRs). In the field of drug development, cell membrane receptors act as selective sites for drug action and account for 50% of all drug targets; GPCR activity-modulating drugs account for 30% of the most commonly used 100 drugs (400 billion dollars, 9% of the total drug market). GPCRs are therefore among the most important targets for new drug development.

[0005] GPCRs share common structural features. They have seven hydrophobic transmembrane domains, each 20-30 amino acids long, connected by hydrophilic amino acid sequences of various lengths. The receptor has an extracellular N-terminus, while the C-terminus is located in the cytoplasm. GTP-binding proteins (G proteins) act as mediators, transmitting the signal generated by the binding of a hormone or other chemical ligand that stimulates the GPCR to intracellular effectors. Upon ligand binding, the intracellular domains of the GPCR undergo a conformational change to allow the receptor to interact with the G protein, which in turn activates intracellular signal transducers, such as adenylyl cyclase, phospholipase C or ion channels. The system generates a signal cascade, in which many secondary transducers act in response to the binding of one ligand to the GPCR. The cell uses this mechanism to detect changes in the extracellular environment and respond appropriately to the change. Overall, endogenous ligands activate the receptor, with the production of a conformational change that allows binding between the receptor and the G protein. Recent studies of the interactions between proteins have revealed that GPCRs bind to various proteins, such as those containing GRK or SH2 (Src homology 2) domains, as well as the adaptor Grb2 and G proteins, to participate in signal transduction.

[0006] Under normal conditions, signal transduction results in the final outcome of cell activation or inhibition. In a physiological environment, GPCRs exist in equilibrium between their inactive and active states in the cell membrane. Inactive receptors are unable to exert a biological response with the cell signal transduction pathway. Only when they are structurally changed to their active form do the receptors exhibit a biological response via the signal transduction pathway (through the G protein). The receptor can be stabilized in the active form by compounds such as endogenous ligands or drugs. Therefore, functional studies such as the cloning of such a gene family and the identification of new ligands for them have the same meaning as the development of new drug candidates, i.e. siRNAs, antibodies, polypeptides, effectors, inhibitors, agonists, antagonists.

[0007] Development, differentiation, homeostasis, response to stimuli, cell cycle control, and aging and apoptosis in living organisms are primarily the result of selective expression of specific genes within cells. The same is true for disease-related cellular mechanisms. In particular, pathological phenomena such as tumorigenesis are caused by gene mutations that ultimately lead to changes in gene expression.

[0008] ONC201 (7-benzyl-4-(2-methylbenzyl)-l,2,6,7,8,9-hexahydroimidazo[l,2- a]pyrido[3,4-e]pyrimidin-5(lH)-one) is the founding member of a class of anticancer compounds called imipridones, which is in Phase II clinical trials for multiple advanced cancers. Since the discovery that ONC201 is a p53-independent inducer of TRAIL gene transcription, preclinical studies have identified that ONC201 has anti-proliferative and pro-apoptotic effects on a wide variety of tumor cells but not normal cells. The mechanism of action of ONC201 involves engagement of the PERK-independent activation of the integrated stress response, leading to tumor upregulation of DR5 and dual Akt / ERK inactivation, and subsequent Foxo3a activation leading to upregulation of the death ligand TRAIL. ONC201 is orally active with infrequent administration in animal models, elicits sustained pharmacodynamic effects, and is not genotoxic. First-in-human ONC201 clinical trials in advanced, aggressive, refractory solid tumors have confirmed that it is well-tolerated. In summary, the imipridone family, including ONC201 and its chemical analogs, represents a new class of therapeutics. SUMMARY

[0009] In one aspect, provided herein is a compound of Formula (10):

[0010]

[0011] wherein R1and R2are independently selected from the group consisting of H, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, aralkylthio, and acyl radicals. In one embodiment, when R1is CH2Ph, R2is not CH2-(2-CH3-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(2-CH3-Ph) (i.e., ONC201). In one embodiment, R1is CH2Ph and R2is CH2-(2,4-diF-Ph) (i.e., ONC206). In one embodiment, R1is CH2Ph and R2is CH2-(4-CF3-Ph) (i.e., ONC212). In one embodiment, R1is CH2Ph and R2is CH2-(3,4-diF-Ph) (i.e., ONC213). In one embodiment, R1is CH2(3,4-di-Cl-Ph and R2is CH2-(4-CF3-Ph) (i.e., ONC234). In one embodiment, R1is CH2-3-thienyl and R2is CH2-(4-CF3-Ph) (i.e., ONC236).

[0012] In another aspect, provided herein are methods of treating or preventing a disease, disorder, or condition in a subject in need thereof, comprising: administering to the subject in need of such treatment a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (10) or an analog thereof, or a pharmaceutically acceptable salt thereof. In an embodiment, the compound is selected from the group consisting of ONC201, ONC206, ONC212, ONC213, ONC234, and ONC236. In an embodiment, the subject has or is at risk of having a cancer. In an embodiment, the cancer is selected from a central nervous system tumor, a brain tumor, a peripheral nervous system tumor, a pheochromocytoma, a paraganglioma, a neuroendocrine tumor, an Ewing sarcoma, a pancreatic cancer, a prostate cancer, an endometrial cancer, a hematological malignancy, a bone cancer, and a lymphatic system tumor. In an embodiment, the cancer is selected from a meningioma, an ependymoma, a glioma, a neuroblastoma, or diffuse intrinsic pontine glioma. In an embodiment, the cancer is selected from an acute leukemia selected from acute lymphoblastic leukemia, acute myeloid leukemia, myelodysplastic syndrome, or myeloproliferative disease. In an embodiment, the cancer has a histone H3 mutation (e.g., H3.3 K27M mutation) or epigenetically silenced unmethylated O(6)-methylguanine-DNA methyltransferase (MGMT) gene. In an embodiment, the subject has or is at risk of having a psychiatric disorder. In an embodiment, the psychiatric disorder is selected from psychosis, schizophrenia, bipolar disorder, or major depressive disorder. In an embodiment, the subject has or is at risk of having an infection. In an embodiment, the infection is a bacterial infection. In an embodiment, the infection is a Gram-negative bacterial infection. In an embodiment, the infection is a Gram-positive bacterial infection. In an embodiment, the bacterial infection is an infection with a bacterium selected from the group consisting of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. In an embodiment, the bacterial infection is a Staphylococcus infection. In an embodiment, the Staphylococcus infection is a S. aureus infection (e.g., a methicillin-resistant S. aureus (MRSA) infection).

[0013] In another aspect, provided herein are methods of treating or preventing a disease, disorder, or condition in a subject in need of selective modulation of a G protein-coupled receptor (GPCR) or a G protein-coupled receptor (GPCR) signaling pathway. Modulation includes, but is not limited to, agonism, partial agonism, inverse agonism, partial antagonism, antagonism, bivalent modulation, or community modulation. In one embodiment, the method comprises administering to a subject in need of such treatment a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has or is at risk of having a cancer. In one embodiment, the subject has or is at risk of having a psychiatric disorder. In one embodiment, the psychiatric disorder is a psychotic disorder. In one embodiment, the psychiatric disorder is schizophrenia. In one embodiment, the subject has or is at risk of having an infection. In one embodiment, the infection is a bacterial infection. In one embodiment, the infection is a Gram-negative bacterial infection. In one embodiment, the infection is a Gram-positive bacterial infection. In one embodiment, the bacterial infection is an infection with a bacterium selected from the group consisting of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. In one embodiment, the bacterial infection is a staphylococcal infection. In one embodiment, the staphylococcal infection is a Staphylococcus aureus infection (e.g., a methicillin-resistant Staphylococcus aureus (MRSA) infection). In one embodiment, the treatment regimen comprises administration of an effective amount of a therapeutic agent, e.g., a compound of Formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the GPCR is a Class A GPCR. In one embodiment, the GPCR is GPR132, GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7, or a combination thereof. In one embodiment, the GPCR is GPR132 (also known as G2A). In one embodiment, the GPCR is GPR91. In one embodiment, the GPCR is MTNR1A. In one embodiment, the GPCR is CXCR7.

[0014] In another aspect, provided herein are methods of treating or preventing a disease, disorder, or condition in a subject in need of selective modulation of the activity of a dopamine receptor or a member of the dopamine receptor signaling pathway. In one embodiment, the method comprises administering to a subject in need of such treatment a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has or is at risk of having a cancer. In one embodiment, the subject has or is at risk of having a psychiatric disorder. In one embodiment, the psychiatric disorder is psychosis. In one embodiment, the psychiatric disorder is schizophrenia. In one embodiment, the subject has or is at risk of having an infection. In one embodiment, the infection is a bacterial infection. In one embodiment, the infection is a Gram-negative bacterial infection. In one embodiment, the infection is a Gram-positive bacterial infection. In one embodiment, the bacterial infection is an infection with a bacterium selected from the group consisting of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. In one embodiment, the bacterial infection is a staphylococcal infection. In one embodiment, the staphylococcal infection is a Staphylococcus aureus infection (e.g., a methicillin-resistant Staphylococcus aureus (MRSA) infection). In one embodiment, the treatment regimen comprises administration of an effective amount of a therapeutic agent, e.g., a compound of Formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is from the D2-like family of dopamine receptors.

[0015] In another aspect, provided herein are methods of treating or preventing liver fibrosis or liver tissue regeneration, comprising: administering to a subject in need of such treatment a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (10) or a compound of Formula (100) (e.g., TIC-10), or an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is a CXCR7 agonist.

[0016] In another aspect, provided herein are methods of stimulating the immune system (e.g., activating NK cells) in a subject in need thereof, the method comprising: administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is a GPR91 agonist. In one embodiment, the compound is ONC213. In one embodiment, the subject has cancer and the method is a method of cancer immunotherapy. In one embodiment, the subject has a viral infection (e.g., HIV). In one embodiment, the subject has systemic lupus erythematosus. In one embodiment, the method further comprises administering to the subject a vaccine (e.g., a cancer vaccine), and the compound is administered as an adjuvant.

[0017] In another aspect, provided herein are methods of identifying whether a subject having a disorder is likely to respond to a treatment regimen described herein. In one embodiment, the method comprises (i) obtaining a biological sample from the subject; (ii) measuring the expression level of at least one dopamine receptor or G protein-coupled receptor (GPCR) in the sample; (iii) comparing the measured level in the sample to a level of a predetermined standard; and (iv) determining whether the subject is likely to respond to the treatment regimen based on the measured level in the sample to the level of the predetermined standard. In one embodiment, the subject has or is at risk of having a cancer. In one embodiment, the subject has or is at risk of having a psychiatric disorder. In one embodiment, the subject has or is at risk of having an infection. In one embodiment, the treatment regimen further comprises administering an effective amount of a therapeutic agent, e.g., a compound of Formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is from the D2-like dopamine receptor family. In one embodiment, the GPCR is a Class A GPCR. In one embodiment, the GPCR is GPR132, GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7, or a combination thereof. In one embodiment, the GPCR is GPR132, GPR91, MTNR1A, CXCR7, or a combination thereof. In one embodiment, the GPCR is GPR132.

[0018] On the other hand, provided herein is a method for assessing the effectiveness of a treatment regimen described herein, monitoring, or providing a prognosis for a subject with a condition. In one embodiment, the method includes (i) obtaining a biological sample from a subject; (ii) measuring the expression level of at least one dopamine receptor or G protein-coupled receptor (GPCR) in the sample; (iii) comparing the level measured in the sample with the level of a predetermined standard; and (iv) determining a prognosis or determining whether the subject is responsive to the treatment regimen based on the level measured in the sample with the level of a predetermined standard. In one embodiment, the method includes (i) obtaining a biological sample from a subject; (ii) measuring the gene copy number or mutation of at least one dopamine receptor in the sample; (iii) comparing the copy number measured in the sample or the mutation found with the copy number or mutation of a predetermined standard; and (iv) determining whether the subject is responsive to the treatment regimen based on the copy number measured in the sample or the mutation found with the copy number or mutation of a predetermined standard. In one embodiment, the subject suffers from or is at risk of suffering from cancer. In one embodiment, the subject suffers from or is at risk of suffering from a mental disorder. In one embodiment, the subject suffers from or is at risk of suffering from an infection. In one embodiment, the treatment regimen comprises administering an effective amount of a therapeutic agent, such as a compound of formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is selected from DRD2, DRD2S, DRD2L, and DRD3. In one embodiment, the dopamine receptor is from the D2-like dopamine receptor family. In one embodiment, the GPCR is a class A GPCR. In one embodiment, the GPCR is GPR132, GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7, or a combination thereof.

[0019] On the other hand, provided herein is a method for screening potential therapeutic agents for a disease. In one embodiment, the method includes (i) contacting at least one G protein-coupled (GPCR) receptor with a test molecule suspected of being a therapeutic agent for a disease; (ii) measuring the binding affinity, interaction, or signaling of the test compound to the GPCR; and (iii) comparing the binding affinity, interaction, or signaling of the test molecule to a predetermined threshold. In one embodiment, the GPCR modulation or GPCR signaling modulation of the test molecule is equivalent to or greater than the threshold value, indicating a therapeutic agent for the disease. In one embodiment, the disease is cancer. In one embodiment, the predetermined threshold value is GPCR modulation or GPCR signaling modulation of a therapeutic agent, such as a compound of formula (10) or a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the GPCR is a class A GPCR. In one embodiment, the GPCR is GPR132. In one embodiment, the GPCR is GPR132, GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7, or a combination thereof. In one embodiment, the GPCR is GPR132. In one embodiment, the GPCR is GPR91. In one embodiment, the GPCR is MTNR1A. In one embodiment, the GPCR is CXCR7.

[0020] On the other hand, provided herein is a method for screening potential therapeutic agents for a disease. In one embodiment, the method comprises (i) contacting at least one dopamine receptor with a test molecule suspected of being a therapeutic agent for a disease; (ii) measuring the binding affinity, interaction or signal transduction of the test molecule with the at least one dopamine receptor; and (iii) comparing the binding affinity or interaction of the test molecule with a predetermined threshold. In one embodiment, the modulation of the dopamine receptor by the test molecule is equivalent to or greater than the threshold value indicating a therapeutic agent for the disease. In one embodiment, the disease is cancer. In one embodiment, the dopamine receptor is a member of the D2-like dopamine receptor family. In one embodiment, the predetermined threshold is the modulation of the dopamine receptor or dopamine receptor signal transduction by a therapeutic agent, such as a compound of formula (10) or a pharmaceutically acceptable salt thereof, or an analog thereof.

[0021] In another aspect, provided herein are methods of screening for potential therapeutic agents for a disorder. In one embodiment, using a processor, the method comprises (i) using a computational docking method to simulate the binding or interaction, if any, of one or more three-dimensional structures (conformations) of a test molecule suspected to be a therapeutic agent for a disorder to a three-dimensional structure of at least one dopamine receptor or model; (ii) using the computational method to assess the binding affinity or interaction of the test molecule structure to the structure of at least one dopamine receptor or model; and (iii) using the computational method to compare the binding affinity or interaction of the test molecule to a predetermined threshold, wherein a modulation of the dopamine receptor by the test molecule equivalent to or greater than the threshold indicates the test molecule as a therapeutic agent for the disorder. In one embodiment, the disorder is cancer. In one embodiment, the dopamine receptor is a member of the D2-like dopamine receptor family.

[0022] In another aspect, provided herein are methods of treating a subject having a disorder. In one embodiment, the method comprises administering an effective amount of a therapeutic agent targeting at least one dopamine receptor or G protein-coupled receptor (GPCR). In one embodiment, the therapeutic agent is a neutralizing agent. In one embodiment, the therapeutic agent is an antagonist of the receptor. In one embodiment, the therapeutic agent is an agonist of the receptor. In one embodiment, the therapeutic agent is a competitive inhibitor of the receptor relative to dopamine. In one embodiment, the therapeutic agent is a non-competitive inhibitor of the receptor relative to dopamine. In one embodiment, the therapeutic agent is selective for the D2-like dopamine receptor family relative to the Dl -like dopamine receptor family. In one embodiment, the subject has or is at risk of having a cancer. In one embodiment, the subject has or is at risk of having a psychiatric disorder. In one embodiment, the subject has or is at risk of having an infection. In one embodiment, the dopamine receptor is a member of the D2-like dopamine receptor family. In one embodiment, the GPCR is a class A GPCR. In one embodiment, the GPCR is GPR132. In one embodiment, the GPCR is GPR91. In one embodiment, the GPCR is MTNR1A. In one embodiment, the GPCR is CXCR7. In one embodiment, the GPCR is GPR132, GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7, or a combination thereof. In one embodiment, the therapeutic agent is a monoclonal antibody (e.g., chimeric or humanized monoclonal antibody), a polyclonal antibody (e.g., chimeric or humanized polyclonal antibody), or a bispecific antibody. In one embodiment, the therapeutic agent is a drug or active agent conjugated to an antibody, e.g., an anti-cancer agent. In one embodiment, the therapeutic agent is a radioconjugated antibody or a small molecule-conjugated antibody. In one embodiment, the therapeutic agent is a vector expressing a recombinant antibody to the dopamine receptor or GPCR. In one embodiment, the therapeutic agent is a fusion protein or peptide targeting the dopamine receptor or GPCR. In one embodiment, the therapeutic agent is an siRNA, shRNA, or antisense oligonucleotide targeting the dopamine receptor or GPCR. In one embodiment, the dopamine receptor or GPCR is targeted by CRISPR interference.

[0023] On the other hand, provided herein are methods for treating subjects with illness and evaluating treatment efficacy. In one embodiment, the method includes (i) treating the subject according to the methods described herein, and (ii) evaluating the efficacy of treatment as described herein. In one embodiment, the subject suffers from or is at risk of suffering from cancer. In one embodiment, the treatment regimen includes administering an effective amount of a therapeutic agent, such as a compound of formula (10), a pharmaceutically acceptable salt thereof, or the like. In one embodiment, based on the measured gene expression level or gene copy number or the mutation found, the therapeutic agent dosage, the frequency of administration of the compound (e.g., compound of formula (10)), or both are selected or adjusted. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The foregoing summary of the invention and the following detailed description of embodiments will be better understood when read in conjunction with the accompanying drawings. However, it should be understood that the invention is not limited to the precise arrangements and instrumentalities shown. In the accompanying drawings:

[0025] Figure 1 : Antagonism of ONC201 on dopamine receptors (DRD1, DRD2S, DRD2L, DRD3, DRD4 and DRD5).

[0026] Figure 2 Soluble prolactin detected by ELISA in peripheral blood of patients with advanced solid tumors at baseline and after a single ONC201 dose (PO 125-625 mg) is shown. Post-treatment sampling time points included 6 hours, 1 day, 2 days, 7 days, and 21 days after treatment.

[0027] Figure 3 : Tumor type sensitivity of the Gene Set of Drug Sensitivity in Cancer (GDSC) cell line collection. Mean sensitivity was estimated by averaging the IC values ​​as determined by cell viability assays performed 72 hours after treatment. 50 The numbers above the bars indicate the number of cell lines per tumor type.

[0028] Figure 4A -D: ONC201 is a selective DRD2 antagonist. ( Figure 4A ) Agonism of orphan or known GPCRs or antagonism of known GPCRs using an arrestin recruitment reporter assay (10 μM ONC201). Figure 4B ) Antagonism of ONC201 against ligand-stimulated dopamine receptors using an arrestin recruitment reporter assay. Figure 4C ) using inhibitory proteins to recruit reporters or ( Figure 4D ) Schild analysis of ONC201 antagonism of DRD2L using a cAMP modulation reporter.

[0029] Figure 5A -F: ONC201 antagonism of DRD2 is highly specific among GPCRs and other cancer drug targets. Figure 5A ) GPCR antagonism using an arrestin recruitment reporter assay (10 μM ONC201). Figure 5B ) inhibitory protein recruitment of reporter molecules or ( Figure 5C ) cAMP modulation reporter molecules resulting in dopamine competition for ONC201-mediated antagonism of DRD2L. Figure 5D ) Antagonistic or agonistic effects of ONC201 (2 or 20 μM) on nuclear hormone receptors were determined using a nuclear translocation reporter assay. Figure 5E )ONC201 (1 μM) inhibits the enzymatic activity of kinases in vitro. ( Figure 5F ) DRD2L antagonistic activity of ONC201 or a biologically inactive linear isomer of ONC201 using an arrestin recruitment reporter assay.

[0030] Figure 6 GBM cell lines with higher DRD2 expression are more responsive to ONC201. (A) Inhibition of NCI60 GBM cell lines as a function of ONC201 concentration. (B) Log ONC201 GI for each GBM cell line 50 (M) relative to DRD2 expression. 2 =0.8707.

[0031] Figure 7 ONC201 exhibits superior selectivity for DRD2 among GPCRs compared to other DRD2 antagonists such as risperidone.

[0032] Figure 8 : ONC201 has higher selectivity for tumor cells than the antipsychotic DRD2 antagonist thioridazine.

[0033] Figure 9 Optimization of ONC201 inhibition of DRD2 calcium flux. HEK-293T cells were transfected with expression constructs for wild-type DRD2 (A) or a control GPCR (B). DRD2-specific calcium flux inhibition was investigated at ONC201 concentrations between 100 pM and 100 μM in the presence of 0.1 and 1 nM dopamine. ONC201 at 100 μM completely inhibited dopamine-induced calcium flux in DRD2 but had no effect on the control GPCR.

[0034] Figure 10: Comparison of DRD2 inhibitors. DRD2-specific calcium flux inhibition was investigated using the inhibitors spiperone (squares), haloperidol (triangles) and ONC201 (circles) at a concentration range under 1 nM dopamine. The non-inhibitor values (shown as 10 -11 M) were used to normalize the data of each assay to 100% activity.

[0035] Figure 11 : Identification of DRD2 residues critical for dopamine-induced calcium flux. (A) Dopamine-induced calcium flux was determined as previously described under 1 nM dopamine across the DRD2 alanine scan library. Data represent the mean of three experiments. Mutant clones were considered to have a defective calcium flux if they exhibited a flux value that was less than 2 standard deviations (AV-2SD) from the average calcium flux value of the entire library. (B) Positions of the 28 mutant residues identified are indicated on the DRD3 crystal structure (green spheres) (PDB id 3PBL; Chien, E.Y. et al. (2010) Science 330: 1091-5). The D2R / D3R antagonist, eticlopride, is shown in cyan.

[0036] Figure 12 : Identification of DRD2 residues critical for ONC201 inhibition of dopamine-induced calcium flux. (A) Dopamine-induced calcium flux was determined as previously described under 1 nM dopamine but in the presence of 100 μΜ ONC201 across the DRD2 alanine scan library. Data represent the mean of three experiments normalized to the value of the flux value with wild-type DRD2. Mutant clones were considered to be critical for ONC201 inhibition if they exhibited a flux value that was greater than 2 standard deviations (AV+2SD) from the average calcium flux value of the entire library. (B) Positions of the 8 mutant residues identified are indicated on the DRD3 crystal structure (red spheres).

[0037] Figure 13 : Reference compound (+) butaclamol and test compound ONC201 dihydrochloride successfully competed for 3 H] methylspiperone with IC 50 values of 2.5 nM and 21 μΜ, respectively.

[0038] Figure 14 : ONC201 dihydrochloride K on and K off determination against DRD2S receptors.

[0039] Figure 15: Compound activity with selected GPCR and orphan GPCR biosensor assays. Compounds were tested in both agonist and antagonist mode with the desired GPCR and orphan GPCR biosensor assays. For agonist assays, data were normalized to the maximum and minimum responses observed in the presence of control ligand and vehicle. For antagonist assays, data were normalized to the maximum and minimum responses observed in the presence of EC 80 Ligand and vehicle. The following EC 80 Concentrations: CCR4 inhibitor protein: 0.0078 mM CCL22; CHRM2 inhibitor protein: 26 mM acetylcholine; and MC4R inhibitor protein: 0.0026 mM melanotan II.

[0040] Figure 16 : ONC206 and ONC212 exhibit anticancer efficacy across various tumor types in the NCI60 panel of cancer cell lines. ONC203 is an inactive negative control.

[0041] Figure 17 : ONC206 is an iloperidone with improved DRD2 antagonism. ONC206 is an analog of ONC201 that exhibits superior antagonism of the D2-like dopamine receptor family and maintains highly selective antagonism of D2-like dopamine receptors compared to other antipsychotic drugs such as haloperidol.

[0042] Figure 18 : Bone cancer is more responsive to ONC206 than ONC201.

[0043] Figure 19 : Ewing’s sarcoma is the most responsive subtype of bone cancer to ONC206.

[0044] Figure 20 : In 14 of 16 Ewing’s sarcoma cell lines, ONC206 exhibits anticancer efficacy in the nanomolar range. In all cell lines, ONC206 exhibits superior efficacy to ONC201.

[0045] Figure 21 : Iloperidone ONC212 targets an orphan GPCR. It is a highly selective agonist of the orphan GPCR tumor suppressor GPR132 and it does not bind to DRD2.

[0046] Figure 22 : ONC212 induces cell death in cancer cells (HCT116) but not normal cells (MRC5) at nanomolar concentrations.

[0047] Figure 23: ONC212 induced integrated stress response and inhibited Akt / ERK phosphorylation at nanomolar concentrations and at an earlier time point compared to ONC201.

[0048] Figure 24 : ONC212 demonstrated oral and intraperitoneal anticancer efficacy in xenograft mouse models of colorectal and breast cancer.

[0049] Figure 25 : Leukemias were more responsive to ONC212 than ONC201.

[0050] Figure 26 : ONC212 demonstrated anticancer efficacy in the nanomolar range (superior efficacy compared to ONC201) in 55 leukemia cell lines, regardless of cell line subtype.

[0051] Figure 27 : GPCRs were agonized or antagonized (>50%) by 9 of the emicidenes tested. Emicidenes selectively target rhodopsin-like Class A GPCRs.

[0052] Figure 28 : Case study of a subject with recurrent glioblastoma (Example 16). (A) Tumor size (%) relative to baseline of total tumor burden in the subject. One cycle is 3 weeks. (B) Comparative MRI scans of one of the 2 malignant lesions at baseline, 21, 27, and 36 weeks after ONC201 initiation.

[0053] Figure 29A - C: ONC212 demonstrated anticancer effects in acute myeloid leukemia (AML) cell lines. Figure 29A Comparison of cell viability of MV411 AML cells treated with ONC212 or cytarabine. Figure 29B Comparison of cell viability of MOLM14, MV411 AML cells, MRC5 lung fibroblasts, and Hs27a bone marrow cells treated with ONC212. Figure 29C Cell viability of MOLM14 and MV411 AML cells treated with ONC212 (250 nM) at 4, 8, 24, 48, 72, and 96 h.

[0054] Figure 30: Efficacy of ONC212 in an ONC201-resistant AML xenograft model (MV411 AML cells (5 x 10 6 subcutaneously implanted in the flank of athymic nude mice). ONC212 and ONC201 were administered orally (PO) as indicated. Tumor volume (A and B) and body weight (C) were measured at the indicated days (n = 10). * indicates p < 0.05 relative to vehicle.

[0055] Figure 31 ONC206 efficacy in an Ewing sarcoma xenograft model (MHH-ES-1 Ewing sarcoma cells (5 × 10 6 ) were implanted subcutaneously into the flanks of athymic nude mice. ONC206 (PO) and methotrexate (IV) were administered as indicated on days 1 and 13. Tumor volume (A) and body weight (B) were measured on the indicated days (n=4).

[0056] Figure 32 : GPCR profile of ONC213 (10 μM) using a β-arrestin recruitment reporter assay.

[0057] Figure 33 ONC213 exhibited in vitro anticancer efficacy in HCT116 / RPMI8226 cancer cells similar to ONC212, but showed lower in vitro toxicity against normal cells than ONC212.

[0058] Figure 34 : GPCR profile of ONC237 (10 μM) using a β-arrestin recruitment reporter assay.

[0059] Figure 35 : GPCR profile of ONC236 (10 μM) using a β-arrestin recruitment reporter assay.

[0060] Figure 36 : GPCR profile of ONC234 (10 μM) using a β-arrestin recruitment reporter assay.

[0061] Figure 37 : GPCR profile of ONC201 linear isomer (TIC-10) (10 μM) using a β-arrestin recruitment reporter assay.

[0062] Figure 38 : The number of GPCR hits of several imipenems. DETAILED DESCRIPTION

[0063] Scientific terms used herein are intended to have the meanings commonly understood by those of ordinary skill in the art. These terms are found in various standard reference works and are used illustratively, and include J. Sambrook and D. W. Russell, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press; Third Edition, 2001; F. M. Ausubel, Ed., Short Protocols in Molecular Biology, Current Protocols; Fifth Edition, 2002; B. Alberts et al., Molecular Biology of the Cell, Fourth Edition, Garland, 2002; D. L. Nelson and M. M. Cox, Lehninger Principles of Biochemistry, Fourth Edition, W. H. Freeman & Company, 2004; Engelke, D. R., RNA Interference (RNAi): Nuts and Bolts of RNAi Technology, DNA Press LLC, Eagleville, PA., 2003; Herdewijn, P. (ed.), Oligonucleotide Synthesis: Methods and Applications, Methods in Molecular Biology, Humana Press, 2004; A. Nagy, M. Gertsenstein, K. Vintersten, R. Behringer, Manipulating the Mouse Embryo: A Laboratory Manual, Third Edition, Cold Spring Harbor Laboratory Press; December 15, 2002, ISBN-10: 0879695919; Kursad Turksen (ed.), Embryonic stem cells: methods and protocols, Methods Mol Biol.2002; 185, Humana Press; Current Protocols in Stem Cell Biology, ISBN: 9780470151808, and U.S. Patent No. 8,673,923. The contents of each of the foregoing references are incorporated herein by reference in their entirety.

[0064] The term“substituted” means that any one or more hydrogen atoms on the designated atom is replaced with the selection of indicated groups, provided that the designated atom’s normal valence is not exceeded, and that the substitution results in a stable compound. When the substituent is a ketone (i.e., =0), 2 hydrogens on the atom are replaced. Ketone aromatic substituents are not present on aromatic moieties. A ring double bond is a double bond formed between two adjacent ring atoms (e.g., C=C, C=N, or N=N).

[0065] When a variable (e.g., R4) occurs more than one time in a constituent or genus of compounds, its definition in each occurrence is independent of its definition in every other occurrence. Thus, for example, if a group is shown to be substituted with 0-3 R4moieties, then the group can optionally be substituted with up to three R4moieties, and at each occurrence R4is independently selected from the definition of R4. Also, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0066] When an atom or chemical moiety is followed by a range of numbers in subscript (e.g., C 1-6 ), it will be understood to encompass all numbers within the range as well as all intervening ranges. For example,“C 1-6 alkyl” means to include alkyl groups having 1, 2, 3, 4, 5, 6, 6-1, 1-5, 1-4, 1-3, 1-2, 2-6, 2-5, 2-4, 2-3, 3-6, 3-5, 3-4, 4-6, 4-5, and 5-6 carbons.

[0067] The term“alkyl” includes both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, C 1-6 alkyl” is meant to include C1, C2, C3, C4, C5, and C6alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl sec-butyl, t-butyl, n-pentyl, sec-pentyl, neopentyl, and n-hexyl. In some instances, straight- chain or branched-chain alkyl groups have six or fewer carbon atoms in their backbone (e.g., C1-C6for straight-chain, C3-C6for branched-chain), in other instances, straight-chain or branched-chain alkyl groups have four or fewer carbon atoms. Likewise, cycloalkyl groups have three to eight carbon atoms in their ring structure, in other instances, cycloalkyl groups have five or six carbon atoms in their ring structure. Most preferred are C 1-6alkyl, especially ethyl, methyl, isopropyl, isobutyl, n-pentyl, n-hexyl and cyclopropylmethyl.

[0068] The term "substituted alkyl" means an alkyl group as defined above, substituted with one, two or three substituents selected from the group consisting of halogen, -OH, alkoxy, -NH2, -N(CH3)2, -C(=O)OH, trifluoromethyl, -C≡N, -C(=O)O(C1-C4)alkyl, -C(=O)NH2, -SO2NH2, -C(=NH)NH2, -NO2, preferably with one or two substituents selected from the group consisting of halogen, -OH, alkoxy, -NH2, trifluoromethyl, -N(CH3)2and -C(=O)OH, more preferably selected from halogen, alkoxy and -OH. Examples of substituted alkyl groups include, but are not limited to, 2,2-difluoropropyl, 2-carboxycyclopentyl and 3-chloropropyl.

[0069] Unless otherwise specified, "lower alkyl" is an alkyl group, as defined above, but having from one to six carbon atoms in its main chain structure, preferably one to four. "Lower alkenyl" and "lower alkynyl" have chain lengths of 2-6 carbon atoms, preferably 2-4 carbon atoms.

[0070] "Alkenyl" includes unsaturated aliphatic groups of similar length and possible substitution of the above-mentioned alkyl groups, but containing at least one double bond. For example, the term "alkenyl" includes straight-chain alkenyl (e.g., ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl), branched-chain alkenyl, cyclic alkenyl (e.g., alicyclic) (e.g., cyclopropenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl- or alkenyl-substituted cycloalkenyl, and cycloalkyl- or cycloalkenyl-substituted alkenyl. In some instances, straight-chain or branched-chain alkenyl groups have six or fewer carbon atoms in their main chain (e.g., straight-chain C2-C6, branched-chain C3-C6). Likewise, cycloalkenyl groups can have three to eight carbon atoms in their ring structure, and in some embodiments, cycloalkenyl groups have five or six carbons in the ring structure. The term "C2-C6" includes alkenyl groups containing 2-6 carbon atoms. The term "C3-C6" includes alkenyl groups containing 3 to 6 carbon atoms.

[0071] "Alkynyl" includes unsaturated aliphatic groups of similar length and possible substitution of the above-mentioned alkyl groups, but containing at least one triple bond. For example, "alkynyl" includes straight-chain alkynyl (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl), branched-chain alkynyl, and cycloalkyl- or cycloalkenyl-substituted alkynyl. In some embodiments, straight-chain or branched-chain alkynyl groups have six or fewer carbon atoms in their main chain (e.g., straight-chain C2-C6, branched-chain C3-C6). The term "C2-C6" includes alkynyl groups containing 2-6 carbon atoms. The term "C3-C6" includes alkynyl groups containing 3 to 6 carbon atoms.

[0072] The term "cycloalkyl" refers to monocyclic or polycyclic non-aromatic radicals in which each atom (i.e., backbone atom) forming the ring is a carbon atom. In some instances, the cycloalkyl group is saturated or partially unsaturated. In other instances, the cycloalkyl group is fused with an aromatic ring. Cycloalkyl groups include groups having from 3 to 10 ring atoms. Examples of cycloalkyl groups include, but are not limited to, the following moieties:

[0073]

[0074] Monocyclic cycloalkyl groups include, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Bicyclic cycloalkyl groups include, but are not limited to, tetralinyl, indanyl, and tetrahydrofurfuryl. Polycyclic cycloalkyl groups include adamantane and norbornane. The term cycloalkyl includes "unsaturated non-aromatic carbocyclic" or "non-aromatic unsaturated carbocyclic" which refer to non-aromatic carbocyclic rings as defined herein which contain at least one carbon-carbon double bond or one carbon-carbon triple bond.

[0075] The term "cycloalkylalkyl" refers to an alkyl group substituted with a cycloalkyl group. Example cycloalkylalkyl groups include cyclopropylalkyl, cyclohexylalkyl.

[0076] The term "heterocycloalkyl" refers to non-aromatic heterocyclic rings in which one or more of the ring-forming atoms is a heteroatom, such as an O, N, or S atom. Heterocycloalkyl groups include monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) ring systems as well as spirocyclic rings. Example heterocycloalkyl groups include morpholino, thiomorpholino, piperazinyl, tetrahydrofuranyl, tetrahydrothiophenyl, 2,3-dihydrobenzofuranyl, 1,3-benzodioxolanyl, benzo-1,4-dioxanyl, piperidinyl, pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, and imidazolidinyl. Also included in the definition of heterocycloalkyl are moieties which can have one or more aromatic rings fused (i.e., sharing a bond) to the non-aromatic heterocyclic ring, such as quinolinyl, isoquinolinyl, and benzo derivatives of heterocycles. Heterocycloalkyl groups having one or more fused aromatic rings are attached through either the aromatic or non-aromatic portion. Also included in the definition of heterocycloalkyl are moieties in which one or more of the ring-forming atoms can be substituted with 1 or 2 oxygen or sulfur groups. In some instances, the heterocycloalkyl group has from 1 to about 20 carbon atoms, and in other instances from about 3 to about 20 carbon atoms. In some instances, the heterocycloalkyl group contains from 3 to 20, from 3 to 2 about 14, from 3 to 27, or from 5 to 26 ring-forming atoms. In some instances, the heterocycloalkyl group has from 1 to 24, from 1 to 23, or from 1 to 22 heteroatoms. In some instances, the heterocycloalkyl group contains from 0 to 23 double bonds. In some instances, the heterocycloalkyl group contains from 0 to 22 triple bonds.

[0077] The term "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl group. An example heteroaralkyl group is pyridylmethyl.

[0078] The term "aryl" refers to a monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) aromatic hydrocarbon, such as phenyl, naphthyl, anthryl, phenanthryl. In some cases, aryl groups have 6 to 20 carbon atoms.

[0079] The term "aryl" refers to a monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) aromatic hydrocarbon, such as phenyl, naphthyl, anthryl, phenanthryl. In some cases, aryl groups have 6 to 20 carbon atoms.

[0080] The term "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl group. An example heteroaralkyl group is pyridylmethyl.

[0081] The term "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl group. An example heteroaralkyl group is pyridylmethyl.

[0082] The term "halogen" or "halo" refers to a fluorine (F), chlorine (Cl), bromine (Br), or iodine (I) atom; preferably F, Cl, or Br; more preferably F or Cl. The term "perhalo" refers to a moiety in which all hydrogens are replaced by halogens.

[0083] The term "haloalkyl" refers to an alkyl moiety having one or more carbons of the hydrocarbon backbone replaced by a halogen. C1-C6 haloalkyl includes straight chain or branched chain alkyl groups having six or fewer backbone atoms and halogen atoms replacing one or more of the backbone carbons' hydrogens.

[0084] The term "alkoxy" or "alkoxy" includes substituted and unsubstituted alkyl, alkenyl, and alkynyl groups covalently linked to an oxygen atom. C1-C6alkoxy refers to moieties having six or fewer carbons in the hydrocarbon backbone. Examples of alkoxy (or alkoxy radicals) include methoxy, ethoxy, isopropoxy, propoxy, butoxy, and pentoxy. Preferred are (C1-C3)alkoxy, especially ethoxy and methoxy. Examples of substituted alkoxy include haloalkoxy.

[0085] The term "hydroxy" or "hydroxyl" includes groups having the formula -OH or -O -

[0086] The term "pharmaceutically acceptable salt" refers to a derivative of a compound in which the parent compound is modified by converting an existing acid or base moiety to its salt form. Pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids. The pharmaceutically acceptable salts include the conventional nontoxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, these salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or a mixture of the two; usually, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts can be found, e.g., in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, PA, 1985, p. 1418, Journal of Pharmaceutical Science, 66, 2 (1977), and P. H. Stahl and C. G. Wermuth, eds., Handbook of Pharmaceutical Salts: Properties, Selection and Use, 2nd Revised and Expanded Edition, Weinheim / Zurich: Wiley-VCH / VHCA (2011), the entire contents of which are incorporated herein by reference.

[0087] ​Examples of suitable inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid or hydrobromic acid, while examples of suitable organic acids include carboxylic acids, sulfonic acids or sulfonic acids, such as acetic acid, tartaric acid, lactic acid, propionic acid, glycolic acid, malonic acid, maleic acid, fumaric acid, tannic acid, succinic acid, alginic acid, benzoic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, cinnamic acid, mandelic acid, citric acid, maleic acid, salicylic acid, trifluoroacetic acid, 3-aminosalicylic acid, ascorbic acid, aconitic acid, nicotinic acid, isonicotinic acid, oxalic acid, gluconic acid, amino acids, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid or naphthalene-2-sulfonic acid. Examples of suitable inorganic bases include sodium hydroxide, potassium hydroxide and ammonia, while examples of suitable organic bases include amines, such as tertiary amines, for example trimethylamine, triethylamine, pyridine, N,N-dimethylaniline, quinoline, isoquinoline, a-methylpyridine, β-methylpyridine, γ-methylpyridine, quinaldine or pyrimidine.

[0088] The term "antibody" includes the native biological form of the structure constituting an antibody. In most mammals, including humans and mice, this form is a tetramer and consists of two identical pairs of immunoglobulin chains, each pair having one light chain and one heavy chain, each light chain comprising an immunoglobulin domain V L and C L , each heavy chain comprising immunoglobulin domains V H , Cγ1, Cγ2 and Cγ3. In each pair, the light and heavy chain variable regions (V L and V H ) together are responsible for binding to the antigen, the constant regions (C L , Cγ1, Cγ2 and Cγ3, in particular Cγ2 and Cγ3) are responsible for antibody effector functions. In some mammals, for example in camels and llamas, full-length antibodies can consist of only two heavy chains, each heavy chain comprising immunoglobulin domains V H , Cγ2 and Cγ3. "Immunoglobulin (Ig)" herein refers to a protein consisting of one or more polypeptides essentially encoded by an immunoglobulin gene. Immunoglobulins include, but are not limited to, antibodies. Immunoglobulins can have many structural forms, including full-length antibodies, antibody fragments and individual immunoglobulin domains including V H , Cγ1, Cγ2, Cγ3, V L and C L .

[0089] Based on the amino acid sequences of the constant domains of the heavy chains, intact antibodies can be assigned to different "classes". There are five major classes of intact antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into "subclasses" (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known and described generally (see, for example, Basic and Clinical Immunology (3rd ed.), Stites and Terr eds., 1980, Chapters 1 and 2).

[0090] The terms "antibody" or "antigen binding fragment" refer to intact molecules and functional fragments thereof, such as Fab, scFv-Fc bivalent molecules, F(ab')2, and Fv capable of specific interaction with a desired target. In some instances, the antigen binding fragment comprises:

[0091] (1) Fab, the fragment of the antibody molecule that contains a single antigen-binding site, which results from digestion with the enzyme papain to yield an intact light chain and a portion of one heavy chain;

[0092] (2) Fab', the fragment of the antibody molecule that can be obtained by treating whole antibody with the enzyme pepsin, followed by reduction, to yield an intact light chain and a portion of one heavy chain; two Fab' fragments are obtained per antibody molecule;

[0093] (3) (Fab')2, the fragment of the antibody that can be obtained by treating whole antibody with the enzyme pepsin without subsequent reduction; F(ab')2 is a dimer of two Fab' fragments held together by two disulfide bonds;

[0094] (4) Fv, a genetically engineered fragment containing the complete coding region of the light chain and the complete coding region of the heavy chain;

[0095] (5) single chain antibody ("SCA"), a genetically engineered molecule containing the variable region of the light chain and the variable region of the heavy chain linked together by a short synthetic linker; and

[0096] (6) scFv-Fc, generated by fusing a single chain Fv (scFv) with a hinge region and Fc region from an immunoglobulin (Ig), such as IgG.

[0097] In one embodiment, the antibody provided herein is a monoclonal antibody. In one embodiment, the antigen binding fragment provided herein is a single chain Fv (scFv), a diabody, a tandem scFv, a scFv-Fc bivalent molecule, a Fab, a Fab', a Fv, a F(ab')2, or an antigen binding scaffold (e.g., an affibody, a unibody, an anticalin, a DARPin, a Knottin).

[0098] The term "bind" or grammatical equivalents means that a composition has affinity for one another, either directly or indirectly. "Specific binding" is the case where two molecules bind selectively to one another. A specific example of specific binding occurs between an antibody and an antigen. In general, specific binding can be distinguished from non-specific binding when the dissociation constant (K D ) is less than about 1 x 10 -5 M or less than about 1 x 10 -6 M or 1 x 10 -7 M. Specific binding can be detected by, for example, ELISA, immunoprecipitation, co-precipitation, with or without chemical cross-linking, and two-hybrid assays. Appropriate controls can be used to distinguish between "specific" and "non-specific" binding. "Affinity" is the strength of the binding interaction of two molecules, for example, an antigen and its antibody, for one another, and for antibodies and other molecules with more than one binding site, is defined as the binding strength of the ligand at one given binding site. While noncovalent linkage of a ligand to an antibody or other molecule is generally not as strong as covalent linkage, "high affinity" means that the affinity constant (K a ) of the ligand binding to the antibody or other molecule is greater than 10 4 M -1 , typically 10 5 -10 11 M -1 ; this is determined by inhibition ELISA or equivalent affinity determined by Scatchard plot or K a / dissociation constant as the reciprocal of K d .

[0099] The term "selective" with respect to binding, inhibition, stimulation, or modulation refers to preferential binding, inhibition, stimulation, or modulation of a first activity over a second activity, respectively (e.g., preferential binding of one receptor over another; preferential inhibition over other receptors; or preferential inhibition of a mutant over wild-type or vice versa). In some cases, the selectivity is greater than two-fold, greater than five-fold, greater than ten-fold, greater than fifty-fold, greater than 100-fold, or greater than 1000-fold selective for a desired molecular target or pathway over an undesired molecular target or pathway. In some cases, a compound binds to a first molecular target or affects a pathway at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least 100-fold, or at least 1000-fold more than a second target or pathway under the same conditions. It will be appreciated that in preferred embodiments, there will be any of the foregoing amounts of selectivity for binding to a D2-like dopamine receptor family or member thereof over a Dl -like dopamine receptor family or member thereof. In vitro or in vivo activity of a molecular target or pathway can be measured by any suitable reproducible means.

[0100] The term "modulate" means to "stimulate" or "inhibit" the activity of a molecular target or pathway. For example, a composition modulates the activity of a molecular target or pathway if it stimulates or inhibits the activity of the target or pathway by at least 10%, at least about 20%, at least about 25%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 90%, at least about 95%, at least about 98%, or about 99% or more relative to the activity of the molecular target or pathway in the presence of the composition under the same conditions but in the absence of the composition only. In another example, a composition modulates the activity of a molecular target or pathway if it stimulates or inhibits the activity of the target or pathway by at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least 100-fold relative to the activity of the molecular target or pathway in the presence of the composition under the same conditions but in the absence of the composition only. The activity of a molecular target or pathway can be measured by any reproducible means. For example, the activity of a molecular target or pathway can be measured in vitro or in vivo by a suitable assay known in the art for measuring activity. A control sample (not treated with the composition) can confer a relative activity value of 100%.

[0101] In one embodiment, the antibody, antigen binding fragment, or affinity tag binds its target with a Kd in the range of 0.1 nM to 10 mM, 0.1 nM to 1 mM, or 0.1 nM. D binds its target. In one embodiment, the antibody, antigen binding fragment, or affinity tag binds its target directly. In one embodiment, the antibody, antigen binding fragment, or affinity tag binds its target indirectly, for example, as a secondary antibody that binds to an antibody that binds the target. D binds its target. In one embodiment, the antibody, antigen binding fragment, or affinity tag binds its target directly. In one embodiment, the antibody, antigen binding fragment, or affinity tag binds its target indirectly, for example, as a secondary antibody that binds to an antibody that binds the target.

[0102] The word "label" refers to a compound or composition that is conjugated or fused directly or indirectly to an agent, such as a nucleic acid probe or an antibody, and that facilitates detection of the agent to which it is conjugated or fused. The label itself can be detectable (e.g., a radioisotope or a fluorescent label), or, in the case of an enzymatic label, can catalyze chemical changes to a substrate compound or composition that are detectable.

[0103] The term "probe" refers to a synthetic or biologically manufactured nucleic acid that contains a specific nucleotide sequence that hybridizes to a target nucleic acid sequence under stringent conditions. The term "labeled probe," "nucleic acid probe operably linked to a detectable label," or "nucleic acid strand operably linked to a detectable label" refers to a probe prepared with a label moiety or "detectable label" for detection. The label moiety is attached at the 5' end, 3' end, internally, or a combination thereof. That is, a probe can be attached to multiple label moieties. The preferred moiety is a recognition label, such as a fluorophore. The labeled probe can also contain multiple different nucleic acid sequences, each labeled with one or more label moieties. Each label moiety can be the same or different. It can be beneficial to have different probes (e.g., nucleic acid sequences) each labeled with a different label moiety. This can be accomplished by having a single distinguishable moiety on each probe. For example, probe A is attached to X moiety and probe B is attached to Y moiety. Alternatively, probe A is attached to X and Y moieties and probe B is attached to Z and W moieties. Alternatively, probe A is attached to X and Y moieties and probe B is attached to Y and Z moieties. All of the above probes "A" and "B" would be distinguishable and uniquely labeled.

[0104] A "tissue sample" refers to a collection of similar cells, preferably containing nucleated cells with chromatin material, obtained from a tissue of a subject or patient. The four major human tissues are (1) epithelium; (2) connective tissue, including blood vessels, bone, and cartilage; (3) muscle tissue; and (4) nervous tissue. The tissue sample source can be from a fresh, frozen, and / or preserved organ or tissue sample or biopsy or aspirated solid tissue; blood or blood components; a body fluid such as cerebrospinal fluid, amniotic fluid, peritoneal fluid, or interstitial fluid; cells from a subject at a certain time of gestation or development. The tissue sample can be primary cells or cultured cells or cell lines. The tissue sample can contain compounds not naturally mixed with the tissue in nature, such as preservatives, anticoagulants, buffers, fixatives, nutrients, or antibiotics. A "section" of a tissue sample refers to an individual portion or piece of a tissue sample, such as a thin slice of tissue or cells cut from a tissue sample. Multiple tissue sections can be taken and analyzed. A "cell line" refers to a permanently established cell culture that will proliferate given fresh culture medium and space.

[0105] Detection methods

[0106] In various aspects, provided herein are methods of detecting or measuring a target receptor (e.g., a dopamine receptor or a GPCR) in a biological sample. The target is detected by contacting the sample with a target detection reagent, e.g., an antibody or fragment thereof, and a labeling reagent. The presence or absence of the target is detected by the presence or absence of the labeling reagent. In some cases, the sample is contacted with the target detection reagent and the labeling reagent simultaneously, e.g., the detection reagent is a primary antibody and the labeling reagent is a fluorescent dye conjugated thereto. Alternatively, the biological sample is contacted with the target detection reagent and the labeling reagent sequentially, e.g., the detection reagent is a primary antibody and the labeling reagent comprises a secondary antibody. For example, the sample is incubated with the detection reagent, in some cases with the labeling reagent, under conditions that allow for the formation of a complex between the detection reagent (and the labeling reagent) and the target. Following complex formation, the sample is optionally washed one or more times to remove unbound detection reagent (and labeling reagent). When the sample is further contacted with a labeling reagent that specifically binds to the detection reagent bound to the target, the sample can be optionally washed one or more times to remove unbound labeling reagent. The presence or absence of the target in the sample is then determined by detecting the labeling reagent.

[0107] The methods described herein provide for detecting multiple targets in a sample. Multiple targets are identified by contacting a biological sample with other detection reagents using the described methods, and then using other labeling reagents specific for the other detection reagents.

[0108] A detectable moiety, i.e., a detectable label, is a substance used to facilitate the identification and / or quantification of a target. Detectable moieties are either directly observed or measured or indirectly observed or measured. Detectable moieties include, but are not limited to, radioactive labels that can be measured with a radiation counter; pigments, dyes, or other chromophores that can be observed visually or measured with a spectrophotometer; spin labels that can be measured with a spin label analyzer; and fluorescent moieties, where the output signal is generated by exciting the appropriate molecular adduct and can be visualized by excitation with light that is absorbed by the dye or can be measured with a standard fluorometer or imaging system. Detectable moieties can be luminescent substances, such as phosphors or fluorophores; bioluminescent substances; chemiluminescent substances, where the output signal is generated by chemical modification of the signaling compound; metal-containing substances; or enzymes, where an enzyme-dependent secondary signal generation occurs, such as the formation of a colored product from a colorless substrate. Detectable moieties can also take the form of chemical or biochemical or inert particles, including colloidal gold, microspheres, quantum dots, or inorganic crystals such as nanocrystals or phosphors. The term detectable moiety or detectable label can also refer to a "tag" or hapten that can selectively bind a labeling molecule, such that the labeling molecule is used to generate a detectable signal upon subsequent addition. For example, biotin, imino-biotin, or desulfo-biotin can be used as a tag, which is then bound to avidin or streptavidin conjugates of horseradish peroxidase (HRP), which is then detected using a chromogenic substrate (e.g., tetramethylbenzidine) or a fluorescent substrate such as Amplex Red or Amplex Gold (Molecular Probes, Inc.). Similarly, the tag can be a hapten or antigen (e.g., digitoxin), and an enzymatic, fluorescent, or radiolabeled antibody can be used to bind the tag. Numerous labels are known to those skilled in the art, including but not limited to particles, fluorescent dyes, haptens, enzymes, and their chromogenic, fluorescent, and chemiluminescent substrates.

[0109] Fluorophores are chemical moieties that exhibit a maximum absorption beyond 280 nm and retain their spectral properties when covalently linked in a labeling reagent. Fluorophores include pyrene, anthracene, naphthalene, acridine, stilbene, indole or benzindole, oxazole or benzoxazole, thiazole or benzothiazole, porphyrin, cyanine, perylene, 4-amino-7-nitrobenzo-2-oxa-l,3-diazole (NBD), carbocyanine, carbostyryl, salicylate, anthranilate, azulene, pyridine, quinoline, boron-dipyrromethene, xanthene, oxazine or benzoxazine, carbazine, phenalene, coumarin, benzofuran, and benzophenalene, and derivatives thereof. Oxazines include tetrazolium, aminooxazinone, diaminooxazine, and benzoid analogs thereof.

[0110] When the fluorophore is an xanthene, the fluorophore can be fluorescein, a p- methoxyaminophenol, or a rhodamine. Fluoresceins include benzo- or dibenzo- fluoresceins, heminaphtho- or naphtho-fluoresceins. Similarly, p-methoxyaminophenols include heminaphthorhodols. Alternatively, the fluorophore is an xanthene which is bound via a single covalent bond at the 9-position of the xanthene. Preferred xanthenes include derivatives of 3H-xanthene-6-ol-3-one, derivatives of 6-amino-3H-xanthene-3-one, or derivatives of 6-amino-3H-xanthene-3-imine. Fluorophores include xanthenes (p-methoxyaminophenols, rhodamines, fluoresceins and derivatives thereof), coumarins, cyanines, pyrenes, oxazines, and boron-dipyrromethene. In addition, the fluorophore can be a sulfonated xanthene, a fluorinated xanthene, a sulfonated coumarin, a fluorinated coumarin, and a sulfonated cyanine. The choice of fluorophore in a labeling reagent will determine the absorption and fluorescence emission properties of the labeling reagent. Physical properties of fluorophore labels including spectral characteristics (absorption, emission and Stokes shift), fluorescence intensity, lifetime, polarization, and photobleaching rates can be used to distinguish one fluorophore from another.

[0111] Typically, the fluorophore contains one or more aromatic or heteroaromatic rings which are optionally substituted with one or more of a variety of substituents including halo, nitro, cyano, alkyl, perfluoroalkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, aralkyl, acyl, aryl or heteroaryl ring systems, benzo, or other substituents typically found on fluorophores known in the art.

[0112] Preferably, the detection moiety is a fluorescent dye. Fluorescent dyes include, for example, Fluorescein, Rhodamine, Texas Red, Cy2, Cy3, Cy5, Cy0, Cy0.5, Cy1, Cy1.5, Cy3.5, Cy7, VECTOR Red, ELF TM (Enzyme-labeled fluorescence), FluorX, Calcein, Calcein-AM, CRYPTOFLUOR TM 'S, Orange (42 kDa), Tangerine 35 kDa), Gold (31 kDa), Red (42 kDa), Crimson (40 kDa), BHMP, BHDMAP, Br-Oregon, Lucifer Yellow, Alexa dye family, N-(6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)hexanoyl) (NBD), BODIPY TM , dipyrrometheneboron difluoride, Oregon Green, MITOTRACKER TMRed, DiOC7(3), DiIC18, Phycoerythrin, Phycobiliproteins BPE (240 kDa) RPE (240 kDa) CPC (264 kDa) APC (104 kDa), Spectrum Blue, Spectrum Aqua, Spectrum Green, Spectrum Gold, Spectrum Orange, Spectrum Red, NADH, NADPH, FAD, Infra-Red (IR) Dye, Cyclic GDP-Ribose (cGDPR), Calcofluor White, Tyrosine and Tryptophan. Many fluorophores can also act as chromophores, so they are also preferred chromophores.

[0113] In addition to fluorophores, enzymes are also found to be useful as detectable moieties. Enzymes are ideal detectable moieties because amplification of the detectable signal can be achieved, resulting in increased sensitivity of the assay. Enzymes do not produce a detectable response by themselves, but when they are contacted with the appropriate substrate, they break down the substrate so that the transformed substrate produces a fluorescent, colorimetric, or luminescent signal. Enzymes amplify the detectable signal because one enzyme on a labeled reagent can result in the transformation of multiple substrates into a detectable signal. This is advantageous when there is a small amount of target present in the sample or when there is no fluorophore present that would produce a comparable or stronger signal than the enzyme. However, fluorophores are preferred because they do not require additional assay steps, thus reducing the total time to complete the assay. Enzyme substrates are selected to produce a preferred measurable product, such as colorimetric, fluorescent, or chemiluminescent. Such substrates are widely used in the art.

[0114] Preferred colorimetric or fluorescent substrates and enzyme combinations use oxidoreductases such as horseradish peroxidase and substrates such as 3,3'-diaminobenzidine (DAB) and 3-amino-9-ethylcarbazole (AEC) which produce distinctive colors (brown and red, respectively). Other colorimetric oxidoreductase substrates that produce detectable products include, but are not limited to: 2,2-amino-bis(3-ethyl ethylbenzothiazoline-6-sulfonic acid) (ABTS), o-phenylenediamine (OPD), 3,3',5,5'-tetramethylbenzidine (TMB), o-dianisidine, 5-aminosalicylic acid, 4-chloro-l-naphthol. Fluorescent substrates include, but are not limited to, homovanillic acid or 4-hydroxy-3-methoxyphenylacetic acid, reduced phenoxazine and reduced benzothiazine, including Amplexe Red reagent and variants thereof, and reduced xanthene, including dihydrofluorescein and dihydorhodamine, the latter including dihydorhodamine 123. Peroxidase substrates that are tyramides represent a unique class of peroxidase substrates that can be inherently detectable prior to enzyme action, but are "fixed in place" by the action of peroxidase in a process described as tyramide signal amplification (TSA). These substrates are widely used to label targets in samples that are cells, tissues, or arrays for subsequent detection by microscopy, flow cytometry, optical scanning, and fluorimetry.

[0115] Other colorimetric (and in some cases fluorescent) substrates and enzyme combinations use phosphatases such as acid phosphatase, alkaline phosphatase, or recombinant forms of such phosphatases in combination with colorimetric substrates such as 5-bromo-6-chloro-3-indolyl phosphate (BCIP), 6-chloro-3-indolyl phosphate, 5-bromo-6-chloro-3-indolyl phosphate, p-nitrophenyl phosphate, or o-nitrophenyl phosphate, or fluorescent substrates such as 4-methylumbelliferyl phosphate, 6,8-difluoro-7-hydroxy-4-methylcoumarinyl phosphate (DiFMUP) fluorescein diphosphate, 3-O-methylfluorescein phosphate, resorcinol phosphate, 9H-(l,3-dichloro-9,9-dimethylindolin-2-one-7-yl)-phosphate (DDAO phosphate), or ELF 97, ELF 39, or related phosphate ester conjugates.

[0116] Glycosidases, particularly β-galactosidase, β-glucuronidase, and β-glucosidase are other suitable enzymes. Suitable colorimetric substrates include, but are not limited to, 5-bromo-4-chloro-3-indolyl β-D-galactopyranoside (X-gal) and similar indolyl galactosides, glucosides, and glucuronides, ortho-nitrophenyl β-D-galactopyranoside (ONPG), and para-nitrophenyl β-D-galactopyranoside. Preferred fluorescent substrates include resorufm β-D-galactopyranoside, fluorescein di-galactoside (FDG), fluorescein di-glucuronide, and structural variants thereof, 4-methylumbelliferyl β-D-galactopyranoside, carboxyumbelliferyl β-D-galactopyranoside, and fluorinated coumarin β-D-galactopyranoside. Other enzymes include hydrolytic enzymes such as cholinesterase and peptidases, oxidases such as glucose oxidase and cytochrome oxidase, and reductases with known suitable substrates.

[0117] Enzymes and suitable substrates therefor that produce chemiluminescence are preferred for certain assays. These include, but are not limited to, luciferase and aequorin in both native and recombinant form. Chemiluminescent substrates for phosphatases, glycosidases, and oxidases, such as those containing stable dioxetanes, luminol, isoluminol, and acridinium esters, are also useful. For example, the enzyme is luciferase or aequorin. The substrate is luciferin, ATP, Ca ++ and coelenterazine.

[0118] In addition to enzymes, haptens such as biotin are useful detectable moieties. Biotin is useful because it can further amplify the detectable signal in an enzyme system, and it can be used as a tag in affinity chromatography for isolation purposes. For detection, an enzyme conjugate with affinity for biotin, such as avidin-HRP, is used. Subsequently, a peroxidase substrate is added to produce a detectable signal. Haptens also include hormones, naturally occurring and synthetic drugs, pollutants, allergens, effector molecules, growth factors, chemotactic factors, cytokines, lymphokines, amino acids, peptides, chemical intermediates, or nucleotides.

[0119] In some cases, the detectable moiety is a fluorescent protein. Exemplary fluorescent proteins include green fluorescent protein (GFP), a phycobiliprotein and its derivatives, luciferase, or aequorin. Fluorescent proteins, especially phycobiliproteins, are particularly useful for generating tandem dye-labeled labeling reagents. These tandem dyes comprise a fluorescent protein and a fluorophore to achieve a greater Stokes shift in which the emission spectrum is further away from the absorption spectrum of the fluorescent protein. This is particularly advantageous for detecting small amounts of target in a sample where the emitted fluorescence is maximally optimized, in other words, the fluorescent protein absorbs little of the emitted light. The fluorescent protein and the fluorophore act as an energy transfer pair, where the fluorescent protein emits at the wavelength absorbed by the fluorophore, which then emits at a wavelength further away from the fluorescent protein than would be possible with the fluorescent protein alone. Particularly useful combinations are phycobiliproteins and sulforhodamine fluorophores, or sulfo-cyanine fluorophores; or sulfo-xanthene derivatives. Alternatively, the fluorophore is the energy donor and the fluorescent protein is the energy acceptor.

[0120] Methods of visualizing the detection moiety depend on the label

[0121] In some cases, the sample is illuminated with a selected wavelength of light to produce a detectable optical response, and observed with a means to detect the response. Equipment useful for illuminating fluorescent compounds includes hand-held ultraviolet lamps, mercury arc lamps, xenon lamps, lasers, and laser diodes. These illumination sources are optically integrated into laser scanners, fluorescence microplate readers, or standard or microscopic fluorometers. The degree or location of the signal compared to a standard or expected response indicates whether and to what extent the sample has a given characteristic or desired target.

[0122] The optical response is detected by visual inspection, or by using one of the following devices: a CCD camera, a video camera, photographic film, a laser scanning device, a fluorometer, a photodiode, a quantum counter, an epi-fluorescence microscope, a scanning microscope, a flow cytometer, a fluorescence micro-well plate reader, or by a means to amplify the signal such as a photomultiplier tube. When the sample is examined using a flow cytometer, the examination of it optionally includes sorting the portions of the sample according to their fluorescent response.

[0123] When a label that can be indirectly detected is used, then the illumination typically involves adding a reagent to produce a detectable signal, such as a colorimetric enzyme substrate. Radioactive isotopes are also considered to be indirectly detectable, in that the radioactive isotope is exposed to x-ray film or other mechanism to record and measure the signal, without the need for additional reagents. This is true for some chemiluminescent signals that are observed after exposure to film.

[0124] I. ONC201 (Compound (1)), salts thereof, and synthesis thereof

[0125] Provided herein is ONC201 (Compound (1)) and their pharmaceutically acceptable salts, and their synthesis. ONC201 has a broad anti-cancer activity, low toxicity, including few adverse effects if any, low genotoxicity, and high bioavailability, including oral, in in vitro models, animal models, and human clinical trials. These characteristics make ONC201 and various analogs highly suitable for a variety of applications. ONC201 can be prepared by synthesis as shown in Scheme 1.

[0126]

[0127] Synthesis of ONC201 dihydrochloride salt begins with the commercially available intermediate, N-benzyl-3-carbomethoxy-4-piperidinone hydrochloride, compound (3). In one embodiment, the synthesis includes neutralizing compound (3) with a base (step 1) to produce compound (4), a free base. In one embodiment, compound (3) is neutralized with an inorganic base to produce compound (4). In one embodiment, compound (3) is neutralized with an organic base to produce compound (4). In one embodiment, compound (3) is neutralized in the presence of an alcohol, e.g., n-butanol. In one embodiment, compound (3) is neutralized in the presence of at least one organic solvent, e.g., n-butanol and / or ethyl acetate. In one embodiment, compound (3) is neutralized in the presence of a base and at least one organic solvent, e.g., NaHC03and n-butanol. In one embodiment, compound (3) is neutralized in the presence of n-butanol and triethylamine (Et3N).

[0128] In one embodiment, the synthesis includes reacting compound (4) and compound (5) (step 2) to produce intermediate compound (1). In one embodiment, the reaction in step 2 includes heating compound (4) with compound (5). In one embodiment, the reaction in step 2 includes heating compound (4) and compound (5) at reflux in the presence of a solvent. In one embodiment, the reaction in step 2 includes using a Dean-stark trap to remove water and / or methanol (MeOH) formed in the reaction.

[0129] In one embodiment, ONC201 dihydrochloride salt is synthesized (step 3). In one embodiment, the reaction (step 3) includes treating ONC201 with HC1 in dioxane. In one embodiment, step 3 includes treating ONC201 with 4N HC1 in dioxane. In one embodiment, the synthesis optionally includes recrystallizing the ONC201 di-salt. In a preferred embodiment, ONC201 dihydrochloride salt is synthesized as shown in Scheme 2.

[0130]

[0131] II. TNF-related apoptosis-inducing ligand ("TRAIL")

[0132] TRAIL protein can be assayed in a sample obtained from a subject to detect TRAIL expression induced by the compounds and salts thereof described herein. Immunoassays can be used to assay TRAIL in a sample, including enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunofilter assay (ELIFA), flow cytometry, immunoblotting, immunoprecipitation, immunohistochemistry, immunocytochemistry, luminescence immunoassay (LIA), fluorescence immunoassay (FIA), and radioimmunoassay. Assays can be used to obtain qualitative and / or quantitative results. Specific details of suitable methods for qualitative and quantitative sample assays are described in standard references, including E. Harlow & D. Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1988; F. Breitling & S. Dubel, Recombinant Antibodies, John Wiley & Sons, New York, 1999; H. Zola, Monoclonal Antibodies: Preparation and Use of Monoclonal Antibodies and Engineered Antibody Derivatives, Basics: From Background to Bench, BIOS Scientific Publishers, 2000; B. K. C. Lo, Antibody Engineering: Methods and Protocols, Methods in Molecular Biology, Humana Press, 2003; F. M. Ausubel et al., eds., Short Protocols in Molecular Biology, Current Protocols, Wiley, 2002; S. Klussman, ed., The Aptamer Handbook: Functional Oligonucleotides and Their Applications, Wiley, 2006; Ormerod, M. G.Flow Cytometry: a practical approach, Oxford University Press, 2000; Givan, A.L., Flow Cytometry: first principles, Wiley, New York, 2001; Gorczyca, W., Flow Cytometry in Neoplastic Hematology: morphologic-immunophenotypic correlation, Taylor & Francis, 2006; Crowther, J.R., The ELISA Guidebook (Methods in Molecular Biology), Humana Press, 2000; Wild, D., The Immunoassay Handbook, 3rdedition, Elsevier Science, 2005, and J. Sambrook and D. W. Russell, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 3rdedition, 2001.

[0133] Protocols for assaying and analyzing samples for TRAIL to detect the effect of the pharmaceutical composition are described in U.S. Patent 8,673,923 to El- deiry et al., which is incorporated by reference in its entirety.

[0134] In one embodiment, the TRAIL assay is used to monitor the subject. For example, a sample is obtained from the subject one or more times before and during and / or after treatment with the pharmaceutical composition to assess the effectiveness of the treatment. In another example, a sample is obtained from the subject various times to assess the progress or advancement of the disease or healing. In one embodiment, the death receptors from circulating tumor cells are assayed to see if the treatments described herein increase the amount or type of death receptors.

[0135] Cancers treated using the methods and compositions described herein are characterized by abnormal cell proliferation, including precancerous hyperproliferation, carcinoma in situ, tumors, and metastases. The methods and compositions described herein can be used for prevention, as well as amelioration of signs or symptoms of cancer. Treatment of a subject for cancer includes: preventing, inhibiting, or ameliorating cancer in a subject, e.g., slowing the progression of cancer or lessening or ameliorating signs or symptoms of cancer. Examples of cancers treated using the methods and compositions described herein include breast cancer, CNS cancer, colon cancer, ovarian cancer, prostate cancer, leukemia, lung cancer, and lymphoma.

[0136] III. Compounds of Formula (10) and salts thereof

[0137] In one aspect, provided herein are compounds of Formula (10) and salts thereof and methods of making the same. It will be understood by those skilled in the art that the general principles and concepts described herein in connection with ONC201 (Compound (1)) and salts thereof, including those related to methods and pharmaceutical compositions, apply with equal force to the compounds of Formula (10) and salts thereof.

[0138] In one embodiment, provided herein are compounds of Formula (10):

[0139]

[0140] wherein R1and R2are independently selected from the group consisting of H, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, aralkyl, heteroarylalkyl, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, aralkylthio, and acyl radicals. In one embodiment, R1is CH2Ph and R2is CH2-(2-CH3-Ph) (i.e., ONC201). In one embodiment, R1is CH2Ph and R2is CH2-(2,4-diF-Ph) (i.e., ONC206). In one embodiment, R1is CH2Ph and R2is CH2-(4-CF3-Ph) (i.e., ONC212). In one embodiment, R1is CH2Ph and R2is CH2-(3,4-diF-Ph) (i.e., ONC213). In one embodiment, R1is CH2(3,4-di-Cl-Ph and R2is CH2-(4-CF3-Ph) (i.e., ONC234). In one embodiment, R1is CH2-3-thienyl and R2is CH2-(4-CF3-Ph) (i.e., ONC236).

[0141] In one embodiment, R1and R2are independently selected from the group consisting of H, C 1-4 alkyl, C 1-4 alkylphenyl, C 1-4 alkylphenyl ketone, C 1-4 benzyl-piperazine, C 1-4 alkylthienyl, C1-4 Alkylpyridyl, C 1-4 Alkylisoxazolidinyl, C 1-4 Alkylmorpholinyl, C 1-4 Alkylthiazolyl and C 1-4 Alkylpyrazinyl, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine, C 1-4 Alkylthienyl, C 1-4 Alkylpyridyl, C 1-4 Alkylisoxazolidinyl, C 1-4 Alkylmorpholinyl, C 1-4 Alkylthiazolyl, C 1-4 The alkylpyrazinyl group is optionally C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, R1 and / or R2 are substituted or unsubstituted, arylalkyl or heteroarylalkyl. In one embodiment, heteroarylalkyl is selected from C 1-4 Alkylpyrrolyl, C 1-4 Alkyl furanyl, C 1-4 Alkylpyridyl, C 1-4 Alkyl-1,2,4-thiadiazolyl, C 1-4 Alkylpyrimidinyl, C 1-4 Alkylthienyl, C 1-4 Alkylisothiazolyl, C 1-4 Alkyl imidazolinyl, C 1-4 Alkyl tetrazolyl, C 1-4 Alkylpyrazinyl, C 1-4 Alkylpyrimidinyl, C 1-4 Alkylquinolinyl, C 1-4 Alkylisoquinolinyl, C 1-4 Thioalkylphenyl, C 1-4 Alkylbenzothiophene, C 1-4 Alkylisobenzofuranyl, C 1-4 Alkylpyrazolyl, C 1-4 Alkyl indolyl, C 1-4 Alkylpurine group, C 1-4 Alkylcarbazolyl, C 1-4 Alkylbenzimidazolinyl, C 1-4 Alkylisoxazolyl.

[0142] In one embodiment, R1and / or R2is benzyl optionally substituted on the benzyl ring with one or more of the following: X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, -OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m , and C(O)OR m ; R m and R n are independently selected from H or C1-C4alkyl; and wherein p is an integer from 2 to 20 and X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl.

[0143] In one embodiment, R2is selected from H, CH3, CH2Ph, CH2-(4-CF3-Ph), CH2-(4-F-Ph), CH2-(4-Cl-Ph), CH2-(OCH3-Ph), CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2-(3-thienyl), CH2-2-pyridyl, CH2-4-methyl-2-thiazolyl, CH2-2-pyrazinyl, CH2CH2Ph, CH2CH2(4-N-benzyl-piperazine), CH2-(2,4-diF-Ph), CH2-(3,4-diCl-Ph), CH2-(3,4-diF-Ph), CH2-(3,5-diF-Ph), CH2-((2-CH3)-Ph), CH2CH(OH)Ph, (4-F-Ph)-4-oxobutyl, CH2CH2NHCOOC(CH3)3, CH2CH2CH2NH2, and CD2C6D5. In one embodiment, R2is selected from H, CH3, CH2Ph, CH2-(4-CF3-Ph), CH2-((2-Cl)-Ph), CH2-((2-F)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzyl-piperazine), CH2-(2,4-diF-Ph), CH2-(2,4-diCl-Ph), CH2-(3,4-diCl-Ph), CH2-(3,4-diF-Ph), CH2-(3,5-diF-Ph), CH2-((2-CH3)-Ph), CH2(2-CH3,4-F-Ph), CH2-((4-OCH3)-Ph), CH2-(3-pyridyl), CH2-(3-isoxazolidinyl), CH2CH2-(4-morpholinyl), CH2-(2-F,4-CF3-Ph), CH2CH(OH)Ph, (CH2)3CO-4F-Ph, (4-F-Ph)-4-oxobutyl, CH2CH2NHCOOC(CH3)3, CH2CH2CH2NH2, and CD2C6D5.

[0144] In one embodiment, R1is H. In one embodiment, R1is a substituted or unsubstituted arylalkyl, such as a benzyl (CH2Ph) or phenethyl group. In one embodiment, the arylalkyl is substituted with C 1-4 alkyl, C 1-4 alkoxy, hydroxyl, perhalogenated C 1-4 alkyl, or halo.

[0145] In one embodiment, R2is a substituted or unsubstituted arylalkyl, such as a benzyl or phenethyl group. In one embodiment, the arylalkyl is substituted with C 1-4 alkyl, C 1-4 alkoxy, hydroxyl, perhalogenated C1-4 In one embodiment, the arylalkyl is substituted with one or more substituents selected from halo, CH3, CF3 or OCH3. In one embodiment, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as piperazinylalkyl or morpholinoalkyl. In one embodiment, R2 is substituted or unsubstituted heteroarylalkyl, such as pyridylmethyl or isoxazolidinylmethyl. In one embodiment, heterocycloalkylalkyl or heteroarylalkyl is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halo substituted. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with at least one halo, CH3, CF3 or OCH3 substituent.

[0146] In one embodiment, compound (10) has the structure of formula (80):

[0147]

[0148] where R a1 , R a2 , R a3 , R a4 , R a5 , R b1 , R b2 , R b3 , R b4 and R b5 Each independently selected from H, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 ,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m ,NR m R n ,NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ; R m and R n independently selected from H or C1-C4 alkyl; and wherein p is an integer from 2 to 20 and X is halogen.

[0149] In one embodiment, compound (10) has the structure of formula (90):

[0150]

[0151] wherein R2is as defined above, and wherein R b1 , R b2 , R b3 , R b4 and R b5 are each independently selected from the group consisting of H, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C 2-4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, -OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ; R m and R n are independently selected from H or C 1-4 alkyl; and wherein p is an integer from 2 to 20 and X is a halogen.

[0152] In one embodiment, compound (10) has the structure of formula (40):

[0153]

[0154] wherein R1is as defined above, and wherein R a1 , R a2 , R a3 , R a4 and R a5 are each independently selected from the group consisting of H, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C 2-4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, -OC p H 2p+1 , -OC p X2p+1 , OR m , SR m ,NR m R n ,NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ; R m and R n Independently selected from H or C 1-4 alkyl; p is an integer from 2 to 20; and X is halogen. In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl group, such as a benzyl or phenethyl group. In one embodiment, the arylalkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, the benzyl group is substituted with one or more halo groups. In one embodiment, the benzyl group is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3, and -OCH3. In one embodiment, the benzyl group is substituted with one halo group, such as F, at the ortho or para position. In one embodiment, the benzyl group is substituted with two halo substituents, such as F, at the two meta positions.

[0155] In one embodiment, compound (40) has the structure of compound (45):

[0156]

[0157] where R a1 、R a2 、R a3 、R a4 and R a5 As defined above. In one embodiment, the benzyl group is substituted with one or more halogens. In one embodiment, the benzyl group is substituted with one or more substituents selected from the group consisting of halogen, CH3, CF3, and OCH3. In one embodiment, R a1 or R a5 is a halogen group, such as F. In one embodiment, R a2 and R a3 Both are halogen groups, such as F.

[0158] In one embodiment, compound (10) has the structure of compound (50):

[0159]

[0160] wherein R1 is as defined above, and wherein Rb selected from the group consisting of H, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C 2-4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, -OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m , and C(O)OR m ; R m and R n are independently selected from H or C 1-4 alkyl; and wherein p is an integer from 2 to 20 and X is a halogen, and wherein R a1 , R a2 , R a4 and R a5 are each independently selected from the group consisting of H, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C 2-4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, -OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m , and C(O)OR m ; R m and R n are independently selected from H or C 1-4alkyl; and wherein p is an integer from 2 to 20 and X is a halogen. In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl group, such as a benzyl or phenethyl group. In one embodiment, the arylalkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, R b is selected from halo, CH3, CF3 and OCH3. In one embodiment, R a1 、R a2 、R a4 and R a5 One or more of are selected from halogen, CH3, CF3 and OCH3. In one embodiment, R a1 、R a2 、R a4 and R a5 is H, and R b is selected from halo, CH3, CF3 and OCH3. In one embodiment, R b is a halogen, such as F, and R a1 is CH3. In one embodiment, R b is F or Cl, and R a2 Is F or Cl. In one embodiment, R b In one embodiment, R b is OCH3. In one embodiment, R b and R a1 It's Cl.

[0161] In one embodiment, compound (50) has the structure of compound (55):

[0162]

[0163] where R a1 、R a2 、R a4 、R a5 and R b As defined above. In one embodiment, R b is selected from halo, CH3, CF3 and OCH3. In one embodiment, R a1 、R a2 、R a4 and R a5 One or more of are selected from halogen, CH3, CF3 and OCH3. In one embodiment, R a1 、R a2 、R a4 and R a5 is H, and Rb is selected from halo, CH3, CF3 and OCH3. In one embodiment, R b is a halogen group, such as F, and R a1 is CH3. In one embodiment, R b is F or Cl, and R a2 Is F or Cl. In one embodiment, R b In one embodiment, R b is OCH3. In one embodiment, R b and R a1 It's Cl.

[0164] In one embodiment, compound (10) has the structure of compound (60):

[0165]

[0166] In one embodiment, R1 is H. In one embodiment, R1 is substituted or unsubstituted arylalkyl, such as benzyl or phenethyl groups. In one embodiment, R1 is substituted or unsubstituted heterocycloalkylalkyl or substituted or unsubstituted heteroarylalkyl, such as CH2-(2-thienyl), CH2-(3-thienyl), CH2-4-methyl-2-thiazolyl, CH2-2-pyrazinyl, CH2CH2(4-N-benzylpiperazine), CH2-(3-isoxazolidinyl), CH2-2-pyridyl, CH2-3-pyridyl and CH2CH2-(4-morpholinyl). In one embodiment, arylalkyl is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, benzyl is substituted with alkyl or halo. In one embodiment, benzyl is substituted with one or more halogen. In one embodiment, benzyl is substituted with one or more substituents selected from halo (e.g., F), CH 3 , CF 3 and OCH 3 . In one embodiment, benzyl is substituted with halo, CH 3 , CF 3 and OCH 3 substituents in the para position. In one embodiment, R 1 is fluorophenoxybutyl or hydroxyphenylethyl.

[0167] Scheme 3 illustrates the synthesis of compounds of formula (10):

[0168] Compound (10) (i.e., amisulpride) is synthesized from a substituted piperidinone which is converted by reaction with a substituted aminoimidazoline to give the core compound (10). There are two routes, where the R1substituent is present in the piperidinone (e.g., 68). In this route, (68) is reacted with dimethylcarbonate using sodium hydride in toluene at 80 °C to form the piperidinone ester (69). Commercially available methylthiohydantoin HI salt (63) is reacted with the amine in dioxane at 70 °C to give the R2-substituted amino-imidazoline (64) as its HI salt. Reaction of (64) with the piperidinone ester (69) in 1-butanol with removal of water through a Dean-Stark trap over 3-6 hours gives the trichloro compound (10). In a variation of this scheme, the N-BOC protected piperidinone (61) is converted to the BOC protected compound (65) by the same method, which is treated with HCI in dioxane to remove the BOC group and then converted to 66) with 1 N NaOH and methyl chloride extraction. Subsequent treatment of (66) with a halide (67) or epoxide (70) gives the desired compound (10).

[0169] The crude product can be purified by column chromatography eluting with methyl chloride:methanol or by HPLC using acetonitrile:TFA:H2O to give the final product as the free base or TFA salt. The free base is treated with HCI in dioxane or the TFA salt is lyophilized to give the product (10) as the HCI or TFA salt. Alternatively, the free base can be treated with another inorganic or organic acid to form other salts, typically selected from those known to be pharmaceutically acceptable salts. The salts of compound (10) are typically solids and examples have been crystallized from ethanol or other solvents to give high quality crystals. In the case of compound (1), the tricyclic structure was confirmed by X-ray crystal structure and NMR.

[0170] The compounds described herein can be used to identify molecules (e.g., proteins) that interact with them in a cellular environment, with or without an aminoalkyl linker (e.g., compound (33)). The expression of these binding targets can be used to predict a response to amisulpride or its analogs (i.e., as a biomarker). These compounds can also be used to screen structurally unrelated molecules using competition assays known in the art to identify drugs that can outcompete the target interaction with higher affinity. In addition, these molecules can have improved drug properties or allow other applications by altering drug properties including safety, potency, pharmacokinetics, biodistribution, or metabolism.

[0171] Table 1: Examples of compounds of formula (10)

[0172]

[0173]

[0174] IV. Assessing sensitivity and efficacy of treatment regimens

[0175] Measuring the expression, genetic mutation, or gene copy number of a dopamine receptor or another G protein-coupled receptor (GPCR) can be used to predict a response or sensitivity to a treatment method described herein and to identify subjects who can respond to a treatment method described herein, e.g., treatment with a compound of Formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one aspect, provided herein are methods of identifying whether a subject having a disorder can respond to a treatment regimen described herein. In one embodiment, the method comprises (i) obtaining a biological sample from the subject; (ii) measuring the level of expression of at least one dopamine receptor or G protein-coupled receptor (GPCR) in the sample; (iii) comparing the level measured in the sample to the level of a predetermined standard; and (iv) determining whether the subject can respond to the treatment regimen based on the level measured in the sample to the level of the predetermined standard. In one embodiment, the step of measuring the level of expression of a dopamine receptor or GPCR in the sample comprises the steps of: (i) contacting the sample with an antibody or antigen-binding fragment that specifically binds to the receptor to form a complex of the antibody or antigen-binding fragment and the receptor; and (ii) measuring the amount of the complex. In one embodiment, the subject has or is at risk of having a cancer. In one embodiment, the cancer is a neuro-oncological disorder. In one embodiment, the cancer is a neuroendocrine tumor. In one embodiment, the cancer is selected from the group consisting of a meningioma, ependymoma, glioma, neuroblastoma, or diffuse intrinsic pontine glioma. In one embodiment, the subject has or is at risk of having a psychiatric disorder. In one embodiment, the psychiatric disorder is selected from the group consisting of psychosis, bipolar disorder, or major depressive disorder. In one embodiment, the subject has or is at risk of having an infection. In one embodiment, the infection is a bacterial infection. In one embodiment, the infection is a Gram-negative bacterial infection. In one embodiment, the infection is a Gram-positive bacterial infection. In one embodiment, the bacterial infection is an infection with a bacterium selected from the group consisting of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. In one embodiment, the Gram-positive bacterial infection is a staphylococcal infection. In one embodiment, the staphylococcal infection is a Staphylococcus aureus infection (e.g., a methicillin-resistant Staphylococcus aureus (MRSA) infection). In one embodiment, the treatment regimen comprises administration of an effective amount of a therapeutic agent, e.g., a compound of Formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is from the D2-like dopamine receptor family. In one embodiment, the dopamine receptor is DRD2. In one embodiment, the dopamine receptor is DRD3. In one embodiment, the dopamine receptor is DRD4. In one embodiment, the dopamine receptor is DRD2, DRD3, or both. In one embodiment, the GPCR is a Class A GPCR.In one embodiment, the GPCR is GPR132. In one embodiment, the GPCR is selected from the group consisting of GPR132, GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7, and combinations thereof. In one embodiment, the dopamine receptor is DRD5, the treatment regimen comprises administration of an effective amount of a therapeutic agent, such as a compound of Formula (10) or a pharmaceutically acceptable salt thereof, and the measured expression level of DRD5 in the sample relative to a predetermined standard indicates whether the subject is likely to respond to the treatment regimen.

[0176] In another aspect, provided herein are methods of evaluating the effectiveness of a treatment regimen described herein, monitoring, or providing a prognosis for a subject having a disorder. In one embodiment, the method comprises (i) obtaining a biological sample from the subject; (ii) measuring the expression level of at least one dopamine receptor or G protein-coupled receptor (GPCR) in the sample; (iii) comparing the level measured in the sample to a predetermined standard; and (iv) determining the prognosis or determining whether the subject responds to the treatment regimen based on the level measured in the sample to the level of the predetermined standard. In one embodiment, the step of measuring the expression level of a dopamine receptor or GPCR in the sample comprises the steps of: (i) contacting the sample with an antibody or antigen-binding fragment that specifically binds to the receptor to form a complex of the antibody or antigen-binding fragment and the receptor; and (ii) measuring the amount of the complex. In one embodiment, the method comprises (i) obtaining a biological sample from the subject; (ii) measuring the gene copy number or mutation of at least one dopamine receptor in the sample; (iii) comparing the copy number measured or mutation found in the sample to the copy number and mutation of a predetermined standard; and (iv) determining whether the subject responds to the treatment regimen based on the copy number measured or mutation found in the sample to the copy number or mutation of the predetermined standard. In one embodiment, the subject has or is at risk of having a cancer. In one embodiment, the cancer is a neuro-oncological disorder. In one embodiment, the cancer is a neuroendocrine tumor. In one embodiment, the cancer is selected from the group consisting of meningioma, ependymoma, glioma, neuroblastoma, or diffuse intrinsic pontine glioma. In one embodiment, the subject has or is at risk of having a psychiatric disorder. In one embodiment, the psychiatric disorder is selected from the group consisting of psychosis, bipolar disorder, or major depressive disorder. In one embodiment, the subject has or is at risk of having an infection. In one embodiment, the infection is a bacterial infection. In one embodiment, the infection is a Gram-negative bacterial infection. In one embodiment, the infection is a Gram-positive bacterial infection. In one embodiment, the bacterial infection is an infection with a bacterium selected from the group consisting of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. In one embodiment, the Gram-positive bacterial infection is a staphylococcal infection. In one embodiment, the staphylococcal infection is a Staphylococcus aureus infection (e.g., a methicillin-resistant Staphylococcus aureus (MRSA) infection). In one embodiment, the treatment regimen comprises administration of an effective amount of a therapeutic agent, e.g., a compound of Formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is selected from DRD2, DRD2S, DRD2L, and DRD3. In one embodiment, the dopamine receptor is from the D2-like dopamine receptor family. In one embodiment, the dopamine receptor is from the D1-like dopamine receptor family. In one embodiment, the dopamine receptor is DRD1.In one embodiment, the dopamine receptor is DRD2. In one embodiment, the dopamine receptor is DRD3. In one embodiment, the dopamine receptor is DRD4. In one embodiment, the dopamine receptor is DRD5. In one embodiment, the dopamine receptor is DRD2, DRD3, or both. In one embodiment, the GPCR is a Class A GPCR. In one embodiment, the GPCR is GPR132. In one embodiment, the GPCR is selected from the group consisting of GPR132, GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7, and combinations thereof.

[0177] In one embodiment, the dopamine receptor is DRD5, the treatment regimen comprises administering an effective amount of a compound of Formula (10) or a pharmaceutically acceptable salt thereof, and the expression level of DRD5 measured in the sample relative to a predetermined standard indicates whether the treatment regimen is effective. In one embodiment, the dopamine receptor is DRD5, the treatment regimen comprises administering an effective amount of a therapeutic agent, e.g., a compound of Formula (10) or a pharmaceutically acceptable salt thereof, and a mutation in the DRD5 gene measured in the sample indicates whether the treatment regimen is effective. In one embodiment, the dopamine receptor is DRD5, the treatment regimen comprises administering an effective amount of a therapeutic agent, e.g., a compound of Formula (10) or a pharmaceutically acceptable salt thereof, and a missense mutation Q366R in the DRD5 gene measured in the sample indicates whether the treatment regimen is effective.

[0178] In another aspect, provided herein are methods of identifying whether a subject having a disorder is likely to respond to a treatment regimen described herein. In one embodiment, the method comprises (i) obtaining a biological sample from the subject; (ii) measuring the gene copy number or mutation in at least one dopamine receptor in the sample; (iii) comparing the copy number or mutation found in the sample to the copy number or mutation of a predetermined standard; and (iv) determining whether the subject is likely to respond to the treatment regimen based on the copy number or mutation measured in the sample compared to the copy number or mutation of the predetermined standard. In one embodiment, the subject has or is at risk of having a cancer. In one embodiment, the cancer is a neural tumor disease. In one embodiment, the cancer is a neuroendocrine tumor. In one embodiment, the cancer is selected from the group consisting of meningioma, ependymoma, glioma, neuroblastoma, or diffuse intrinsic pontine glioma. In one embodiment, the subject has or is at risk of having a psychiatric disorder. In one embodiment, the psychiatric disorder is selected from the group consisting of psychosis, schizophrenia, bipolar disorder, or major depressive disorder. In one embodiment, the subject has or is at risk of having an infection. In one embodiment, the infection is a bacterial infection. In one embodiment, the infection is a Gram-negative bacterial infection. In one embodiment, the infection is a Gram-positive bacterial infection. In one embodiment, the bacterial infection is an infection by a bacterium selected from the group consisting of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species. In one embodiment, the Gram-positive bacterial infection is a staphylococcal infection. In one embodiment, the staphylococcal infection is a Staphylococcus aureus infection (e.g., a methicillin-resistant Staphylococcus aureus (MRSA) infection). In one embodiment, the treatment regimen comprises administration of an effective amount of a therapeutic agent, e.g., a compound of Formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is from the D2-like dopamine receptor family. In one embodiment, the dopamine receptor is DRD1. In one embodiment, the dopamine receptor is DRD2. In one embodiment, the dopamine receptor is DRD3. In one embodiment, the dopamine receptor is DRD4. In one embodiment, the dopamine receptor is DRD5. In one embodiment, the dopamine receptor is DRD2, DRD3, or both. In one embodiment, the dopamine receptor is DRD5, the treatment regimen comprises administration of an effective amount of a therapeutic agent, e.g., a compound of Formula (10), or a pharmaceutically acceptable salt thereof, and the mutation in the DRD5 gene measured in the sample indicates whether the subject is likely to respond to the treatment regimen. In one embodiment, the dopamine receptor is DRD5, the treatment regimen comprises administration of an effective amount of a therapeutic agent, e.g., a compound of Formula (10), or a pharmaceutically acceptable salt thereof, and the missense mutation Q366R in the DRD5 gene measured in the sample indicates whether the subject is likely to respond to the treatment regimen.

[0179] In addition, measuring the expression, post-translational modification, or activity level or mutation in eIF2-a, ATF4, CHOP, DR5, or fragmented or total cytokeratin 18 can be used to predict response or sensitivity to the treatment methods described herein or to identify subjects who can respond to treatment with a compound of Formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. Further, measuring the expression, post-translational modification, or activity level or mutation in eIF2-a, ATF4, CHOP, DR5, or fragmented or total cytokeratin 18 can be used to assess or monitor the effectiveness of the treatment methods described herein. In addition, measuring the expression, post-translational modification, or activity level or mutation in eIF2-a, ATF4, CHOP, DR5, or fragmented or total cytokeratin 18 can be used to screen structurally unrelated anti-cancer molecules in vivo, in vitro, or in silico. For example, competition and other assays can be used to identify drugs that are able to out-compete the target interaction with higher affinity, to compare these levels of change to the respective changes produced by a compound of Formula (10) or an analog thereof. Assays can also be performed on living mammalian cells, which more closely approximate the effects of particular serum levels of drugs in vivo, or microsomal extracts prepared from cultured cell lines.

[0180] In one embodiment, the subject has or is at risk of having cancer. In one embodiment, the treatment regimen comprises administration of an effective amount of imipridone, e.g., ONC201, or an analog thereof. In one embodiment, the treatment regimen comprises administration of an effective amount of ONC201. In one embodiment, the treatment regimen comprises administration of an effective amount of a compound of Formula (10). In one embodiment, the compound of Formula (10) is a compound of Formula (40), e.g., a compound of Formula (45). In one embodiment, the compound of Formula (10) is a compound of Formula (50), e.g., a compound of Formula (55). In one embodiment, the compound of Formula (10) is a compound of Formula (80). In one embodiment, the compound of Formula (10) is a compound of Formula (90). In one embodiment, the compound of Formula (10) is a compound of Formula (60). In one embodiment, the analog of compound (1) has a structure selected from the structures of Formula (25), Formula (26), Formula (27), Formula (28), Formula (29), Formula (30), or Formula (31).

[0181] The predetermined standard level can be, for example, an average or median level measured in a sample from a subject. The predetermined standard level can be measured under the same or substantially similar experimental conditions as the sample from the subject. The predetermined standard level can be obtained from a subject who responds to treatment with the chemical, e.g., ONC201, or an analog thereof. In one embodiment, the predetermined standard is obtained from a subject who responds to treatment with a chemical, and the subject can be classified as likely to respond to treatment if the level in the sample from the subject is similar to the level in the standard. The predetermined standard level can be obtained from a subject who does not respond to treatment with a chemical. In one embodiment, the predetermined standard is obtained from a subject who does not respond to treatment with a chemical, and the subject can be classified as likely to respond to treatment if the level in the subject sample is different (e.g., upregulated or downregulated) from the level in the expected standard. The predetermined standard level can be obtained from a normal, healthy subject.

[0182] Immunoassays can be used to determine protein or methyl chemical levels in a sample, including enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunosorbent assay (ELIFA), flow cytometry, immunoblotting, immunoprecipitation, immunohistochemistry, immunocytochemistry, luminescence immunoassay (LIA), fluorescence immunoassay (FIA), and radioimmunoassay. 6 A mRNA methyl chemical level can be obtained by methyl RNA immunoprecipitation (Me-RIP) or other quantitative biochemical assays known in the art.

[0183] Nucleic acid mutations can be determined by any of a number of known methods. For example, a biological sample of an individual can be obtained. Such biological samples include, but are not limited to, bodily (e.g., urine, saliva, plasma, or serum) or tissue samples (e.g., buccal tissue samples or buccal cells). The biological sample can then be sequenced or scanned using known methods. For example, DNA arrays can be used to analyze at least a portion of a subject's genomic sequence. In addition, all or a portion of the genomic sequence information can be used. Such sequences can be determined using standard sequencing methods including chain termination (Sanger dideoxynucleotides), dye terminator sequencing, and SOLID™ sequencing (Applied Biosystems). The entire genomic sequence can be sequenced by restriction enzyme cleavage or shearing (mechanically) into shorter fragments. DNA sequences can also be amplified using known methods such as PCR and vector-based cloning methods (e.g., E. coli). In one embodiment, at least a portion of a subject's genetic material (e.g., DNA, RNA, mRNA, cDNA, other nucleotide bases, or derivatives of these) is scanned or sequenced using, for example, a conventional DNA sequencer or a chip-based technology to determine whether a mutation or copy number variation is present.

[0184] In one aspect, provided herein are methods of identifying and treating subjects having a disorder and likely to respond to a treatment regimen described herein. In one embodiment, the method comprises: (i) identifying whether a subject having a disorder is likely to respond to a treatment regimen described herein; (ii) treating a subject determined to be likely to respond to the treatment regimen with the treatment regimen. In one embodiment, the subject has or is at risk of having a cancer. In one embodiment, the treatment regimen comprises administration of an effective amount of imipridone, e.g., ONC201, or an analog thereof. In one embodiment, the treatment regimen comprises administration of an effective amount of Compound (1). In one embodiment, the treatment regimen comprises administration of an effective amount of a compound of Formula (10). In one embodiment, the compound of Formula (10) is a compound of Formula (40), e.g., a compound of Formula (45). In one embodiment, the compound of Formula (10) is a compound of Formula (50), e.g., a compound of Formula (55). In one embodiment, the compound of Formula (10) is a compound of Formula (80). In one embodiment, the compound of Formula (10) is a compound of Formula (90). In one embodiment, the compound of Formula (10) is a compound of Formula (60). In one embodiment, the analog of Compound (1) has a structure selected from the structure of Formula (25), Formula (26), Formula (27), Formula (28), Formula (29), Formula (30), or Formula (31).

[0185] The predetermined standard level can be, for example, an average or median level measured in a sample from a subject. The predetermined standard level can be measured under the same or substantially similar experimental conditions as the sample from the subject. The predetermined standard level can be obtained from a subject who responds to treatment with imipridone, e.g., ONC201, or an analog thereof. In one embodiment, the predetermined standard is obtained from a subject who responds to treatment with a chemical, and the subject can be classified as likely to respond to treatment if the level in the sample from the subject is similar to the level in the standard. The predetermined standard level can be obtained from a subject who does not respond to treatment with a chemical. In one embodiment, the predetermined standard is obtained from a subject who does not respond to treatment with a compound, and the subject can be classified as likely to respond to treatment if the level in the subject sample is different (e.g., upregulated or downregulated) from the level in the expected standard. The predetermined standard level can be obtained from a normal, healthy subject. An immunoassay can be used to determine the level of the protein in the sample.

[0186] In one aspect, provided herein are methods of treating and assessing the effectiveness of treatment in a subject having a disorder. In one embodiment, the method comprises: (i) treating the subject according to the methods of treatment described herein, (ii) assessing the effectiveness of the treatment as described herein. In one embodiment, the subject has or is at risk of having cancer. In one embodiment, the treatment regimen comprises administration of an effective amount of imipridone, e.g., ONC201, or an analog thereof. In one embodiment, the treatment regimen comprises administration of an effective amount of Compound (1). In one embodiment, the treatment regimen comprises administration of an effective amount of a compound of Formula (10). In one embodiment, the compound of Formula (10) is a compound of Formula (40), e.g., a compound of Formula (45). In one embodiment, the compound of Formula (10) is a compound of Formula (50), e.g., a compound of Formula (55). In one embodiment, the compound of Formula (10) is a compound of Formula (80). In one embodiment, the compound of Formula (10) is a compound of Formula (90). In one embodiment, the compound of Formula (10) is a compound of Formula (60). In one embodiment, the analog of Compound (1) has a structure selected from the structure of Formula (25), Formula (26), Formula (27), Formula (28), Formula (29), Formula (30), or Formula (31).

[0187] Other disorders that can be suitable for the methods described herein include attention deficit disorders, addictions, epilepsy, viral infections, inflammation, neurodegenerative diseases such as Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, amyotrophic lateral sclerosis, cardiovascular diseases such as coronary artery disease, cardiomyopathy, hypertensive heart disease, heart failure, cor pulmonale, arrhythmia, inflammatory heart disease, endocarditis, inflammatory heart disease, myocarditis, valvular heart disease, cerebrovascular disease, peripheral arterial disease, congenital heart disease, rheumatic heart disease, diabetes, and light chain amyloidosis.

[0188] V. Compositions

[0189] In one aspect, provided are pharmaceutical compositions comprising a compound of Formula (10): or Formula (1): and pharmaceutically acceptable salts thereof. In one embodiment, the salt is a pharmaceutically acceptable di-salt of the compound. In one embodiment, the salt is a pharmaceutically acceptable mono- or poly-salt (e.g., di- or tri-salt) selected from the group consisting of hydrochloride, hydrobromide, bisulfate, sulfate, phosphate, fumarate, succinate, oxalate, and lactate, bisulfate, hydroxyl, tartrate, nitrate, citrate, tartrate, carbonate, malate, maleate, fumarate sulfonate, methanesulfonate, formate, acetate, and carboxylate. In one embodiment, the salt is a salt selected from the group consisting of p-toluenesulfonate, benzenesulfonate, citrate, methanesulfonate, oxalate, succinate, tartrate, fumarate, and maleate. In one embodiment, the salt is a salt selected from the group consisting of ammonium, sodium, potassium, calcium, magnesium, zinc, lithium, and / or counterions such as methylamino, dimethylamino, diethylamino, and triethylamino counterions. In one embodiment, the salt is a dihydrochloride or dihydrobromide salt.

[0190] Compound (1) (ONC201) has the same chemical structure revealed by structural analysis (e.g., NMR, X-ray diffraction) of chemical substance NSC 350625 obtained from the National Cancer Institute Developmental Therapeutics Program Repository.

[0191] In one embodiment, the pharmaceutical composition includes a di-salt of ONC201 (e.g., dihydrochloride) or an analog thereof (e.g., imipridone). A salt (e.g., di- or tri-salt) of an ONC201 analog can be prepared from an ONC201 analog, which can be synthesized as described herein, or using standard chemical synthesis methods known to one of ordinary skill in the art.

[0192] In one embodiment, the pharmaceutical composition includes at least one pharmaceutically acceptable carrier. Suitable pharmaceutically acceptable carriers include, but are not limited to, those in the "Handbook of Pharmaceutical Excipients," 7th Ed., Raymond C. Rowe et al., Eds., American Pharmaceutical Association, Washington, USA and Pharmaceutical Press, London; and earlier editions. Exemplary pharmaceutically acceptable carriers, compounds and methods of various dosage forms and modes of administration are well known in the art and are described in detail, for example, in Larry L. Augsburger & Stephen W. Hoag, Eds., Pharmaceutical Dosage Forms: Tablets, London: Informa Healthcare, 2008; and L.V. Allen, Jr. et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 8th Ed., Philadelphia, Pa.: Lippincott, Williams & Wilkins, 2004; A.R. Gennaro, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st edition, 2005, especially Chapter 89; and J.G. Hardman et al., Goodman & Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill Professional, 10th edition, 2001.

[0193] In one embodiment, the pharmaceutical composition is formulated for ocular administration. In one embodiment, the pharmaceutical composition is for topical administration. In one embodiment, the pharmaceutical composition is formulated as an oil drop, ointment or liquid. In one embodiment, the pharmaceutical composition includes a conventional pharmaceutical carrier such as an aqueous, powder or oily base, thickeners.

[0194] In one embodiment, the pharmaceutical composition is a formulation for intravenous administration. In one embodiment, the intravenous formulation comprises a compound of Formula (10) or a pharmaceutically acceptable salt thereof dissolved in a solvent. In one embodiment, the solvent comprises water. In one embodiment, the intravenous formulation comprises the compound or salt thereof at a concentration of about 0.05, about 0.25, about 0.5, about 2.5, about 5, about 25, or about 50 mg / mL. In one embodiment, the intravenous formulation comprises the compound or salt thereof at a concentration of about 0.05, 0.5, or 5 mg / mL to about 1, 10, or 100 mg / mL. In one embodiment, the intravenous formulation comprises the compound or salt thereof at about 0.005%, 0.05%, or about 0.5% to about 0.1%, 1%, or 10%. In one embodiment, the intravenous formulation comprises the compound or salt thereof at about 0.05%, 0.5%, or 5%. In one embodiment, the intravenous formulation comprises a higher or lower concentration of the compound or salt thereof.

[0195] In one embodiment, the intravenous formulation has a pH of about 3. In one embodiment, the formulation is adjusted to pH 3 with a phosphate buffer. In one embodiment, the intravenous formulation comprises dextrose or sodium chloride. In one embodiment, the intravenous formulation comprises the compound or salt thereof at a concentration of about 5 mg / mL and a pH of 3, forming a stable solution. In one embodiment, the intravenous formulation comprises the compound or salt thereof at a concentration of about 5 mg / mL, a pH < 5, forming a stable solution. In one embodiment, the intravenous formulation comprises the compound or salt thereof and one or more antioxidants. In one embodiment, the intravenous formulation comprises a mixture of the mono- and dihydrochloride salts of the compound. In one embodiment, the intravenous formulation comprises the compound or salt thereof at a concentration of about 10 mg / mL as a 1% solution, for example, the intravenous formulation is a solution having a pH of about 3.3. In one embodiment, the pH is less than 4.0.

[0196] In one embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. In one embodiment, suitable pharmaceutically acceptable carriers include aqueous carriers. In one embodiment, the aqueous carrier comprises sterile water. In one embodiment, the formulation comprises dextrose and / or sodium. In one embodiment, the pharmaceutically acceptable carrier comprises an oil.

[0197] In one embodiment, the intravenous formulation comprises ONC201 or an analog thereof or its dihydrochloride salt dissolved in water at 25 mg / mL. In one embodiment, the formulation is adjusted to pH 3 with phosphate buffer. In one embodiment, the formulation comprises dextrose, sodium chloride, or both. In one embodiment, the formulation comprises a higher or lower concentration of the dihydrochloride salt of ONC201 or an analog thereof. In one embodiment, the formulation comprises ONC201 or an analog thereof or its dihydrochloride salt at a concentration of about 5 mg / mL. In one embodiment, the formulation of about 5 mg / mL forms a stable solution and has a pH of 3. In one embodiment, the formulation of about 5 mg / mL has a pH < 5 and forms a stable solution. In one embodiment, the intravenous formulation comprises ONC201 or an analog thereof or its dihydrochloride salt and one or more antioxidants. In one embodiment, the intravenous formulation comprises a mixture of the mono- and dihydrochloride salts of ONC201 or an analog thereof. In one embodiment, the intravenous formulation comprises ONC201 or an analog thereof or its dihydrochloride salt as a 1% solution at a concentration of about 10 mg / mL. For example, the intravenous formulation is a solution having a pH of about 3.3. In one embodiment, the pH is less than 4.0.

[0198] In one embodiment, the intravenous formulation comprises about 0.5% to about 10% (or about 5 mg / mL to about 100 mg / mL) of ONC201 or an analog thereof or its dihydrochloride salt. In one embodiment, the formulation comprises about 5% (or about 50 mg / mL) or more of ONC201 or an analog thereof or its dihydrochloride salt. In one embodiment, the rate of intravenous infusion can be slowed to reduce side effects of ONC201 or an analog thereof or its dihydrochloride salt.

[0199] In one embodiment, the pharmaceutical composition comprises about 0.1-99% of a salt of ONC201 or an analog thereof; and a pharmaceutically acceptable carrier, such as an oil or sterile water or other aqueous carrier. In one embodiment, for oral dosage forms, the composition comprises in the range of about 5% to about 50% of a mono- or di-salt of ONC201 or an analog thereof.

[0200] In one embodiment, the pharmaceutical composition includes an antioxidant. Suitable antioxidants include: ascorbic acid derivatives such as ascorbic acid, erythorbic acid, sodium ascorbate, thioglycerol, cysteine, acetylcysteine, cystine, dithioerythritol, dithiothreitol, glutathione, tocopherols, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), sulfites such as sodium sulfite, sodium bisulfite, acetone sodium bisulfite, sodium metabisulfite, sodium sulfite, sodium formaldehyde sulfoxylate, and sodium thiosulfate, and dihydroguaiaretic acid. It should be noted that antioxidants for aqueous formulations typically include: sodium sulfite, sodium bisulfite, sodium formaldehyde sulfoxylate, and ascorbic acid and combinations thereof, while antioxidants for oil-based solutions, organic solvents include butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and propyl propionate and combinations thereof. In other embodiments, the antioxidant can be one or more of flavonoids, isoflavones, monothioglycerol, L-cysteine, thioglycolic acid, alpha-tocopherol, ascorbyl 6-palmitate, dihydrolipoic acid, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), vitamin E, propyl gallate, beta-carotene, ascorbic acid. Antioxidants can typically be used at about 0.1% to 1.0% by weight, more typically about 0.2%.

[0201] In one embodiment, the pharmaceutical composition comprises imirichene, e.g., ONC201, or an analog thereof or a pharmaceutically acceptable salt thereof and at least one other therapeutic agent. For example, the other therapeutic agent is selected from the group consisting of hormone analogs and anti-hormones, aromatase inhibitors, LHRH agonists and antagonists, growth factor inhibitors, growth factor antibodies, growth factor receptor antibodies, tyrosine kinase inhibitors; antimetabolites; antitumor antibiotics; platinum derivatives; alkylating agents; antimitotic agents S; tubulin inhibitors; PARP inhibitors, topoisomerase inhibitors, serine / threonine kinase inhibitors, tyrosine kinase inhibitors, protein-protein interaction inhibitors, RAF inhibitors, MEK inhibitors, ERK inhibitors, IGF-1R inhibitors, ErbB receptor inhibitors, rapamycin analogs, BTK inhibitors, CRM1 inhibitors (e.g., KPT185), P53 modulators (e.g., Nutlins), anti-angiogenic factors (e.g., atilimumab, aldesleukin, sorafenib, and regorafenib), amifostine, anagrelide, clodronate, felicitin, interferon, interferon alpha, folinic acid, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate, and porfimer, 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-alethine, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxy camptothecin, 16-aza-epothilone B, A105972, A204197, abiraterone, aldesleukin, ivermectin, allovectin-7, altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037, AP-5280, apaziquone, apomine, aranose, arglabin, arzoxifene, atamestane, atrasentan, auristatin PE, AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), ARRY 162, ARRY 438162, ARRY-300, ARRY-142886 / AZD-6244 (selumetinib) 704 / AZD-8330, AR-12, AR-42, AS-703988, AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026 (primasertib) AZD-2014, azacitidine, azanatabine B, azonafide, BAY-43-9006, BAY80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, benzydamide disodium, BCX-1777, BKM-120, blicomycin, BLP-25, BMS-184476, BMS-247550,BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW 2992 (afatinib, Gilotrif), BIBF 1120 (vargagef), BI 836845, BI 2536, BI 6727, BI 836845, BI 847325, BI 853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryostatin-1, bortezomib, brostallicin, busulphan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, capecitabine, canfosfamide, capecitabine, carboxyphthalin, CCI-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, cefotaxime, ceftriaxone, celecoxib, cemiplimab, cemiplimab, CH4987655 / RO-4987655, chloranil, cilengitide, cyclosporin, CDA-II, CDC-394, CKD-602, CKI-27, clofarabine, colchicine, combretastin A4, COT inhibitor, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibody, CP-461, CV-247, cyanomorpholino doxorubicin, cytarabine, D 24851, decitabine, deoxypyridoxine, deoxypyridoxine, deoxyribonucleic acid mycosin, desphospholipid ascorbic acid B, dexamethasone, dexamethasone, diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10, dolastatins, DS-7423, E7010, E-6201, edatrexat, edotreotide, efaproxiral, efomycin B, epratuzumab, epothilone B, eravacycline, ER-86526, erlotinib, ET-18-0CH3, ethynyl cytidine, ethynyl estradiol, exatecan, exatecan mesylate, exemestane, exisulind, fenretinide, filoplafibat, filoplafibat, folic acid, FOLFOX, FOLFOX4, FOLFIRI, gefitinib, gemtuzumab, gefitinib, gefitinib, gefitinib, GFC-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100,gp100 peptide vaccine, GSK-5126766, GSK-690693, GSK-1120212 (trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron, Herceptin, hexamethylmelamine, histamine, homoharringtonine, hyaluronic acid, hydroxyurea, hydroxyprogesterone caproate, ibandronate, ibrutinib, itraconazole, escitalopram, escitalopram, IDN-5109, IGF-1R inhibitor, IMC-1C11, IMC-A 12 (citumumab), immunoglobulin, hemolysin, interferon alfa-2a, interferon alfa-2b, pegylated interferon alfa-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib, ipilimumab, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, kahalidF, ketoconazole, KW-2170, KW-2450, lobaplatin, leflunomide, isoflavone plus, leuprorelin, leuporelin, lexidronam, LGD-1550, linezolid, THB, tylosin, lorestat, ruxolitinib, LU 223651, loxoantrone, LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechlorethamine, MEK inhibitors, MEK-162, methyltestosterone, methylprednisolone, MEDI-573, MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, midostaurin, minodronic acid, mitomycin, mivobulin, MK-2206, MK-0646 (daloizumab), MLN518, motesafam gadolinium, MS-209, MS-275, MX6, neridronate, neratinib, Nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nolatrexed, Norelin, N-acetylcysteine, 06-benzylguanine, oblimersen, omeprazole, oncophage, oncoVEXGM-CSF, ormiplatin, ortataxel, OX44 antibody, OSI-027, OSI-906 (linsitinib), 4-1BB antibody, oxantrazole, estrogen, panitumumab, patupirone, pefirstim, PCK-3145, pEG-005, PF-05197281, PF-05212384, PF-04691502, PHT-427, P-04, PKC412, P54, PI-88, pelitinib, pemetrexed,Pentrix, perifosine, perillyl alcohol, pertuzumab, PI3K inhibitor, PI3K / mTOR inhibitor, PG-TXL, PG2, PLX-4032 / RO-5185426 (vemurafenib)-3603 / RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pivaloyloxymethylbutyrate, heptanoprenone, phentodil O, PKI166, pletrexed, mithramycin, polymalonic acid, porphomicin, prednisone, prednisolone, quinolone, R115777, RAF-265, ramosetron, ranpirnase, RDEA-119 / BAY 869766, RDEA-436, pterostilbemicin analogs, receptor tyrosine kinase (RTK) inhibitors, regorafenib, revimid, RG-7167, RG-7304, RG-7421, RG-7321, RG 7440, rhizobactam, rhu-MAb, relifadine, risedronate, rituximab, rofanumab, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR 109881A, daunorubicin phenylhydrazone, rubitecan, R-flurbiprofen, RX-0201, S-9788, abalabicin, SAHA, sargramostim, satraplatin, SB 408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustin, 4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spirocarboplatin, keratinine, hydroxamic acid, sutent, T 900607, T 138067, TAK-733, TAS-103, tylosin, taparopenem, tylosin, tynosartan, tesaptan, tegafur, temozolomide, tesarofen, temofil, temofilin, testosterone, propionate, tesmilifene, tetrachloroplatinic acid, tetrodotoxin, tizocillin, thalidomide, trastuzumab, teprazole, trovastatin, trovastatin, tramatinib, trans-MID-107, retinoic acid, trastuzumab, tretinoin, tretinoin, triacetyluridine, triapine, triciribine, trimethoprim-sulfamethoxazole, TLK-286TXD 258, tykerb / tyverb, urobilirubin, mindin, varacini, vincristine, virulizumab, WX-UK1, WX-554, Vectibix, Xeloda, XELOX, XL-147, XL-228, XL-281, XL-518 / R-7420 / GDC-0973, XL-765, YM-511, YM-598, ZD-4190, ZD-6474, ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZD1839, ZSTK-474,zoledronat, zosuquidar and combinations thereof.

[0202] In one embodiment, the other therapeutic agent comprises a hormone analogue, an anti-hormone or both selected from the group consisting of tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate, flutamide, nilutamide, bicalutamide, aminoglutethimide, cyproterone acetate, finasteride, buserelin acetate, flurohydrocortisone, fluoxymesterone, medroxyprogesterone acetate, octreotide, and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more LHRH agonists and / or antagonists selected from the group consisting of goserelin acetate, ciotropate acetate, triptorelin pamoate and combinations thereof and wherein the LHRH antagonist is selected from the group consisting of degarelix, cetrorelix, abarelix, ozarelix, and combinations of galic. In one embodiment, the other therapeutic agent comprises one or more growth factor inhibitors selected from the group consisting of inhibitors of platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), insulin-like growth factor (IGF), human epidermal growth factor (HER) and hepatocyte growth factor (HGF). In one embodiment, the other therapeutic agent comprises one or more inhibitors of human epidermal growth factor selected from the group consisting of HER2, HER3 and HER4. In one embodiment, the other therapeutic agent comprises one or more tyrosine kinase inhibitors selected from the group consisting of cetuximab, gefitinib, imatinib, lapatinib and trastuzumab and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more aromatase inhibitors selected from the group consisting of anastrozole, letrozole, liarozole, vorozole, exemestane, atamestane and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more anti-metabolites which are antifolates selected from the group consisting of methotrexate, raltitrexed and pyrimidine analogues. In one embodiment, the other therapeutic agent comprises one or more anti-metabolites which are pyrimidine analogues selected from the group consisting of 5-fluorouracil, capecitabine and gemcitabine. In one embodiment, the other therapeutic agent comprises one or more anti-metabolites which are purine and / or adenosine analogues selected from the group consisting of mercaptopurine, thioguanine, cladribine and pentostatin, cytarabine, fludarabine and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more anti-tumour drugs, doxorubicin, daunorubicin and deoxydoxorubicin, bleomycin-C, bleomycin, dactinomycin, mithramycin, streptozocin and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more platinum derivatives selected from the group consisting of cisplatin, oxaliplatin, carboplatin and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more alkylating agents selected from the group consisting of estramustin, meclorethamine, melphalan, chlorambucil, busulphan, dacarbazin,cyclophosphamide, ifosfamide, temozolomide, nitsoureas, and combinations thereof. In one embodiment, the additional therapeutic agent comprises a nitsourea selected from carmustin, lomustin, thiotepa, and combinations thereof. In one embodiment, the additional therapeutic agent comprises an anti-mitotic agent selected from a vinca alkaloid and a taxane. In one embodiment, the additional therapeutic agent comprises one or more taxanes selected from paclitaxel, docetaxel, and combinations thereof. In one embodiment, the additional therapeutic agent comprises one or more vinca alkaloids selected from vinblastine, vincristine, vinorelbine, and combinations thereof. In one embodiment, the additional therapeutic agent comprises one or more topoisomerase inhibitors that are epipodophyllotoxins. In one embodiment, the additional therapeutic agent comprises one or more epipodophyllotoxins selected from etoposide and teniposide, nimustine, amsacrin, topotecan, irinotecan, mitoxantron, and combinations thereof. In one embodiment, the additional therapeutic agent comprises one or more serine / threonine kinase inhibitors selected from a PDK 1 inhibitor, a B-Raf inhibitor, an mTOR inhibitor, an mTORC1 inhibitor, a PI3K inhibitor, a dual mTOR / PI3K inhibitor, a STK 33 inhibitor, an AKT inhibitor, a PLK 1 inhibitor, a CDK inhibitor, an Aurora kinase inhibitor, and combinations thereof. In one embodiment, the additional therapeutic agent comprises one or more tyrosine kinase inhibitors that are PTK2 / FAK inhibitors. In one embodiment, the additional therapeutic agent comprises one or more protein-protein interaction inhibitors selected from IAP, Mcl-1, MDM2 / MDMX, and combinations thereof. In one embodiment, the additional therapeutic agent comprises one or more rapalogs selected from everolimus, temsirolimus, ridaforolimus, temsirolimus, and combinations thereof. In one embodiment, the additional therapeutic agent comprises one or more agents selected from amifostine, anagrelide, clodronat, filgrastin, interferon, interferon alpha, leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate, and porfimer, and combinations thereof. In one embodiment, the additional therapeutic agent comprises one or more agents selected from 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-alethine, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxy camptothecin, 16-aza-epothilone B, A105972, A204197, abiraterone, aldesleukin, acitretin, allovectin-7, altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037,AP-5280, azaroxin, alitretinoin, aliskiren, arcticitum, arcticitum, arcticitum, artemisitan, atezolizumab, aurostatin PE, ABT-199 (Venetoclax), ABT-263 (Navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganetymab), ARRY 162, ARRY 438162, ARRY-300, ARRY-142886 / AZD-6244 (selumetinib)-704 / AZD-8330, AR-12, AR-42, AS-703988, AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026 (primasertib), AZD-2014, azacytidine, azanator B, azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, bexarotene disodium, BCX-1777, BKM-120, blicomycin, BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW 2992 (afatinib, Gilotrif), BIBF 1120 (vargatef), BI 836845, BI 2536, BI 6727, BI 836845, BI 847325, BI 853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryostatin-1, bortezomib, brostallicin, busulphan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, capecitabine, canfosfamide, capecitabine, carboxyphthalin, CCl-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, cefotaxime, ceftriaxone, celecoxib, cemiplimab, cemiplimab, CH4987655 / RO-4987655, chloranil, cytarabine, cyclosporin, CDA-II, CDC-394, CKD-602, CKI-27, clofarabine, colchicine, combretastin A4, COT inhibitor, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibody, CP-461, CV-247, cyano-morpholino doxorubicin, cytarabine, D 24851,Decitabine, Doxorubicin, Deoxypyrrolnitrin B, Dexamethasone, Dexamethasone, Diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10, dolarazyme, DS-7423, E7010, E-6201, edatrexate, eflornithine, EGFR inhibitor, EKB-569, EKB-509, enzastaurin, enzalomid, enoxacin, epothilone B, epratuzumab, ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine, ethynylestradiol, exatecan, exatecan mesylate, exemestane, exisulind, fenretinide, filgrastim, floxuridine, folic acid, FOLFOX, FOLFOX4, FOLFIRI, fostresant, fluvoxetine, camptothecin, maltitol, gefitinib, gemtuzumab, gimatecan, glucosamide, GCS-100, GDC-0623, GDC-0941 (pictrelisib) 0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100, gp100-peptide vaccine, GSK-5126766, GSK-690693, GSK-1120212 (trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron, herceptin, hexamethylmelamine, histamine, homoharringtonine, hyaluronic acid, hydroxyurea, hydroxyprogesterone caproate, ibandronic acid ibritumomab, ixabepilone, itraconazole, isatuximab, isofosfamide, IGF-1R inhibitor, IMC-1C11, IMC-A12 (cixutumumab), immunoglobulin, hemolysin, interferon alpha-2a, interferon alpha-2b, pegylated interferon alpha-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib, isoproterenol, isofagin B, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, kahalid F, ketoconazole, KW-2170, KW-2450, lobaplatin, leflunomide, leuporelin, lexidronam, LGD-1550, linezolid, lutetium texaphyrin, lomustine, loosxantrone, LU 223651, losoxantrone, LY-S6 AKT1, LY-2780301, mafosfamide, marimastat, mechlorethamine, MEK inhibitor, MEK-162, methyltestosterone, methylprednisolone, MEDI-573,MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, midostaurin, minodronic acid, mitomycin, mivobulin, MK-2206, MK-0646 (dalotuzumab), MLN518, motexafin gadolinium, MS-209, MS-275, MX6, neridronate, neratinib, Nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nolatrexed, norlestrin, N-acetylcysteine, 06-benzylguanine, oblimersen, omeprazole, oncophage, oncoVEX GM-CSF, ormiplatin, ortataxel, OX44 antibody, OSI-027, OSI-906 (linsitinib), 4-1BB antibody, oxantrazole, estrogen, panitumumab, patupilone, pegfilgrastim, PCK-3145, pegfilgrastim, PBI-1402, PBI-05204, PDO325901, PD-1 antibody, PEG- paclitaxel, albumin-stabilized paclitaxel, PEP-005, PF-05197281, PF-05212384, PF-04691502, PHT-427, P-04, PKC412, P54, PI-88, pelitinib, pemetrexed, pentrix, perifosine, pinoresinol, pertuzumab, PI3K inhibitors, PI3K / mTOR inhibitors, PG-TXL, PG2, PLX-4032 / RO-5185426 (vemurafenib), PLX-3603 / RO-5212054, PT-100, PWT-33597, PX-866, platinol, pivaloyloxymethyl butanoic acid, povidone, phenoxychlorohydrin O, PKI166, pertussis, porfiromycin, polylysine, porfimer, prednisone, prednisolone, quinine, quinupristin, R115777, RAF-265, ramucirumab, rapogovab, RDEA-119 / BAY 869766, RDEA-436, rebecamycin analog, receptor tyrosine kinase (RTK) RG-7167, RG-7304, RG-7421, RG-7321, RG7440, rhizoxin, rhu-MAb, linifanib, risedronic acid, rituximab, robatumumab, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR 109881A, pyridazinone, erythromycin, R-flurbiprofen, RX-0201, S-9788, doxorubicin, SAHA, sagopilone, satraplatin, SB 408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustin, seocalcitol, SM-11355, SN-38,SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squaramate, hydroxamic acid, sutent, T 900607, T 138067, TAK-733, TAS-103, tachykantin, tacrolimus, tylosin, tesacolone, tegafur, temozolomide, tesarodine, testosterone, testosterone propionate, tilmivisene, tetrachloroaurate, tetrodotoxin, tizanidine, thalidomide, trastuzumab, enalapril, tomethamb, talampanel, torcetrapib, tomivirine, tomifene, trametinib, trans-MID-107, retinoic acid, trastuzumab, tremelimumab, tretinoin, triacetyluridine, triaziridine, TLK-286 TXD 258, tykerb / tyverb, urocidin, valrubicin, vatalanib, vincristine, vindesine, virulizin, WX-UK1, WX-554, veltuzumab, xeloda, XELOX, XL-147, XL-228, XL-281, XL-518 / R-7420 / GDC-0973, XL-765, YM-511, YM-598, ZD-4190, ZD-6474, ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZD1839, ZSTK-474, zoledronat, zosuquidar, and combinations thereof.

[0203] In one embodiment, the additional therapeutic agent comprises a steroid, including dexamethasone, prednisolone, methylprednisolone, prednisone, hydrocortisone, fludrocortisone, betamethasone, and coicosterone. In one embodiment, the additional therapeutic agent comprises an antiemetic agent, including but not limited to 5-HT3 receptor agonists (e.g., dolasetron, granisetron, ondansetron, tropisetron, palonosetron, and mirtazapine), dopamine agonists (e.g., domperidone, olanzapine, droperidol, haloperidol, chlorpromazine, prochlorperazine, alizapride, prochlorperazine, and metoclopramide), NK1 receptor antagonists (e.g., aprepitant and casopitant), antihistamines (e.g., cyclizine, diphenhydramine, dimenhydrinate, doxylamine, meclizine, promethazine, hydroxyzine), cannabinoids (e.g., cannabis, nabilone, dronabinol, and tetrahydrocannabinol), benzodiazepines (e.g., midazolam and lorazepam), anticholinergics (e.g., hyoscine), trimethobenzamide, ginger, emetrol, propofol, peppermint, muscimol, and capsicum.

[0204] In one embodiment, the additional therapeutic agent comprises an anticancer agent, including a mitotic inhibitor. In one embodiment, the mitotic inhibitor comprises a taxane. In one embodiment, the mitotic inhibitor comprises a taxane selected from paclitaxel and docetaxel.

[0205] In one embodiment, the pharmaceutical composition comprises emilione, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof; and at least one anticancer agent, including one or more of avermectin, aclarubicin, atradazole, aksuline, atifostine, acitretin, allopurinol, atrazine, onychomycin, ipratropium, amifostine, aminoglutarimide, amsacrine, anastrozole, citalopram, arsenic trioxide, asparaginase, aspirin, azacitidine, azetepa, azotomycin, batimastat, benzotelluride, bevacizumab, bicalutamide, bisantrene, bischofide dimesylate, bleomycin, brequinar, bromocriptine ... Amine, busulfan, cactinomycin, capecitabine, carcimetine, carboplatin, carmustine, carboplatin, carmustine, cariprigin, celecoxib, celecoxib, chlorambucil, siromulin, cisplatin, cladribine, cinorbendazole mesylate, cyclophosphamide, cytarabine, dacarbazine, actinomycin D, daunomycin, decitabine, dexamethasone, dezagunine, dezagunine mesylate, diazocine, docetaxel, doxorubicin, droloxamine, droloxifene, drostanolone, duroxamine, edatrexamine, etisulin, edoxacin, enloplatin, oxazolidinone, ebuloside, etoposide, etopostatin, faldolazol, fazarapine, fenretinide, fluorouracil, fludarabine, fluorouracil, flucitabine, fluoxetine, fostamicin, fulvestrant, gemcitabine, Hydroxyurea, daunorubicin, ifosfamide, sarcosine, interleukin II (IL-2, including recombinant interleukin II or rIL2), interferon alfa-2a, interferon alfa-2b, interferon alfa-1, interferon alfa-alpha3, interferon beta-1a, interferon gamma-1b, methadone, melentalline acetate, melphalan, mecamylamine, mercaptopurine, methotrexate, metoprine, meturedepa, mitindomide, mitocarcin, mitocromin, mitocromin, mitomycin, mitomycin, mitomycin, mitoxantrone, mitotane, mitoxantrone, mycophenolic acid, nelarabine, nocodazole, nokaramycin, olanzapine, oxazolidinone, taxane Alcohol, polyethylene glycol, pomomycin, pentomustine, amide, pipobroman, piposulfan, piroxatone hydrochloride, mithramycin, primimane, porphin, porphyromycin, prednimustine, procarbazine, puromycin, pyrrolidone, pyroxine, rogamid, benzoinol, ulinastatin, pyrazole, stavudine, strenomycin, spiramycin, spiramycin, spirosulfate, zosin, sumonifen, temozole, temoxifen, temoporfin, teniposide, tenilolone, tiotropolone, timolol, thioguanine, thiotapa, thiazofuran, tirabasamide, topotecan, toremifene, terilolone, triptorelin, trastuzumab, uracil mustard, vitampin, vilbiprofen,Vinflunine, vinblastine, vincristine sulfate, vindesine, vinflunine, vorbutin, vinorelbine, vorisodine, vezoloxetine, vorozoo, zenaplatin, zenostatin, zoledronic acid, zolithromycin, and combinations thereof.

[0206] Examples of suitable anticancer agents include those described in Goodman and Gilman, The Pharmacological Basis of Therapeutics, 12th edition, edited by Laurence Brunton, Bruce Chabner, Bjorn Knollman, McGrawHill Professional, 2010.

[0207] In some exemplary embodiments, the pharmaceutical composition comprises a salt (e.g., monosalt or disalt) of emilione, such as ONC201, or an analog thereof, and at least one other therapeutic agent, wherein the other therapeutic agent comprises an anti-angiogenic agent, such as bevacizumab. In one embodiment, the anti-angiogenic agent is selected from aflibercept, axitinib, angiostatin, endostatin, a 16 kDa prolactin fragment, laminin peptide, fibronectin peptide, tissue inhibitor of metalloproteinases (TIMPs), or a combination thereof. 1, 2, 3, 4), plasminogen activator inhibitors (PAI-1, -2), tumor necrosis factor alpha, (high dose, in vitro), TGF-β1, interferon (IFN-α, -β, γ), ELR-CXC chemokine, IL-12; SDF-1; MIG; platelet factor 4 (PF-4); IP-10, thrombospondin (TSP), SPARC, 2-methoxyestradiol, proliferator-activated protein kinase, suramin, sorafenib, tirofenib, thalidomide, cortisone, mitochondria, fumagillin (AGM-1470; TNP-470), tamoxifen, tretinoin, CM101, dexamethasone, leukemia inhibitory factor (LIF), hedgehog inhibitors and combinations thereof.

[0208] The pharmaceutical combination can include any desired ratio of the first and second therapeutic agents, so long as a synergistic or synergistic effect still occurs. Synergistic pharmaceutical combinations preferably contain the first and second therapeutic agents in a ratio of about 1:9 to about 9:1. In one embodiment, the synergistic combination contains the first and second therapeutic agents in a ratio of about 1:8 to about 8:1, about 1:7 to about 7:1, about 1:6 to about 6:1, about 1:5 to about 5:1, about 1:4 to about 4:1, about 1:3 to about 3:1, or about 1:2 to about 2:1. In one embodiment, the synergistic combination contains the therapeutic agents in a ratio of approximately 1:1.

[0209] In one embodiment, the second therapeutic agent is selected from allopurinol, arsenic trioxide, azacitidine, bortezomib, bevacizumab, capecitabine, carboplatin, celecoxib, chlorambucil, clofarabine, cytarabine, dacarbazine, daunorubicin hydrochloride, docetaxel, doxorubicin hydrochloride, floxuridine, gemcitabine hydrochloride, hydroxyurea, ifosfamide, imatinib mesylate, ixabepilone, lenalidomide, megestrol acetate, methotrexate, mitotane, mitoxantrone hydrochloride, oxaliplatin, paclitaxel, pralatrexate, romidepsin, sorafenib, streptozotocin, tamoxifen citrate, topotecan hydrochloride, tretinoin, vandetanib, vismodegib, vorinostat, and combinations thereof.

[0210] In one embodiment, the second therapeutic agent comprises a small molecule multikinase inhibitor, such as sorafenib or regorafenib. In one embodiment, the second therapeutic agent comprises a Hedgehog pathway inhibitor, such as vismodegib. In one embodiment, the second therapeutic agent comprises a drug selected from Table 2 below.

[0211] Table 2: Drug Classes

[0212]

[0213]

[0214]

[0215] In one embodiment, the second therapeutic agent comprises a drug that targets tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors. In one embodiment, the second therapeutic agent comprises a recombinant TRAIL or agonist antibody that activates one or more TRAIL receptors. In one embodiment, the second therapeutic agent comprises one or more antibodies or recombinant TRAIL that activate signaling through DR4, DR5, or both. In one embodiment, the second therapeutic agent comprises one or more of AMG-655, LBY-135, mapamumab, lexalimumab, Apomab, and rhApo2L / TRAIL. In one embodiment, the second therapeutic agent comprises an agent selected from the group consisting of camptothecin, 5-FU, capecitabine, cisplatin, doxorubicin, irinotecan, paclitaxel, cisplatin, bortezomib, BH3I-2, rituximab, radiation, triterpenes, sorafenib, gemcitabine, HDAC inhibitors, carboplatin, T-101 (gossypol derivative), ABT-263, ABT-737, and GX-15-070 (obatoclax), vorinostat, cetuximab, panitumumab, bevacizumab, ganitazumab, interferon gamma, sorafenib, XIAP antagonists, Bcl-2 antagonists, and Smac mimetics.

[0216] VI. Dosage

[0217] In one embodiment, the pharmaceutical composition comprises imirichmane, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 40, 50, 60, or 100 mg to about 2000 mg; about 4, 6, 6, or 10 mg to about 200 mg; or about 0.4, 0.5, 0.6, or 1 mg to about 20 mg, where the weights can be based on the compound in its free base form. In one embodiment, the pharmaceutical composition comprises imirichmane, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 50 mg to about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg; from about 5 mg to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, and 200 mg; or about 0.5 mg to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 mg. In one embodiment, the pharmaceutical composition comprises imirichmane, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 40 mg to about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg; from about 4 mg to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg; or about 0.4 mg to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 mg. In one embodiment, the pharmaceutical composition comprises imirichmane, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 60 mg to about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg; about 6 mg to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg; or about 0.6 mg to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg.In one embodiment, the pharmaceutical composition comprises an imiricotide, e.g., ONC201 or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 100 mg to about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg; from about 10 to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg; or about 1 mg to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg. In one embodiment, the pharmaceutical composition comprises an imiricotide, e.g., ONC201 or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 200 mg to about 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg; about 20 mg to about 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg; or about 2 mg to about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg, based on the compound in its free base form. In one embodiment, the pharmaceutical composition comprises an imiricotide, e.g., ONC201 or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 400 mg to about 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg; about 40 mg to about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg; or about 4 mg to about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg, based on the compound in free base form.In one embodiment, the pharmaceutical composition comprises imiricitabine, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of from about 50 mg to about 60, 70, 80, 90, or 100 mg; from about 60 mg to about 70, 80, 90, or 100 mg; from about 70 mg to about 80, 90, or 100 mg, from about 80 mg to about 90, or 100 mg; from about 90 mg to about 100 mg; from about 5 mg to about 6, 7, 8, 9, or 10 mg; from about 6 mg to about 7, 8, 9, or 10 mg; from about 7 mg to about 8, 9, or 10 mg, from about 8 mg to about 9, or 10 mg; from about 9 mg to about 10 mg; from about 0.5 mg to about 0.6, 0.7, 0.8, 0.9, or 1 mg; from about 0.6 mg to about 0.7, 0.8, 0.9, or 1 mg; from about 0.7 mg to about 0.8, 0.9, or 1 mg, from about 0.8 mg to about 0.9, or 1 mg; or from about 0.9 mg to about 1 mg.

[0218] In one embodiment, the pharmaceutical composition comprises imirichne, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 1 mg / kg to about 40 mg / kg; 0.1 mg / kg to about 4 mg / kg; or 0.01 mg / kg to about 0.40 mg / kg. In one embodiment, the pharmaceutical composition comprises imirichne, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, about 1, 2, 3, 4, 5, 6, 7, 8, or 9 mg / kg to about 10, 20, 30, or 40 mg / kg; about 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 mg / kg to about 20, 30, or 40 mg / kg; about 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 mg / kg to about 30 or 40 mg / kg; about 30, 31, 32, 33, 34, 35, 36, 37, 38, or 39 mg / kg to about 40 mg / kg; about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, or 0.9 mg / kg to about 1, 2, 3, or 4 mg / kg; about 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, or 1.9 mg / kg to about 2, 3, or 4 mg / kg; from about 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, or 2.9 mg / kg to about 3 or 4 mg / kg; or from about 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, or 3.9 mg / kg to about 4 mg / kg; about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 mg / kg to about 0.10, 0.20, 0.30, or 0.40 mg / kg; about 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, or 0.19 mg / kg to about 0.20, 0.30, or 0.40 mg / kg; about 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, or 0.29 mg / kg to about 0.30 or 0.40 mg / kg; or about 0.30 mg / kg about 0.30, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, or 0.39 mg / kg to about 0.40 mg / kg.

[0219] In one embodiment, the pharmaceutical composition comprises imirichne, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range from about 37.5 mg / m 2 to about 1500 mg / m 2 , from about 3.75 mg / m2 from about 0.1 mg / m2to about 150 mg / m2 2 from about 0.1 mg / m2to about 150 mg / m2 2 from about 0.1 mg / m2to about 150 mg / m2 2In one embodiment, the pharmaceutical composition comprises eprazinone, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range from about 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995, 1000, 1005, 1010, 1015, 1020, 1025, 1030, 1035, 1040, 1045, 1050, 1055, 1060, 1065, 1070, 1075, 1080, 1085, 1090, 1095, 1100, 1105, 1110, 1115, 1120, 1125, 1130, 1135, 1140, 1145, 1150, 1155, 1160, 1165, 1170, 1175, 1180, 1185, 1190, 1195, 1200, 1205, 1210, 1215, 1220, 1225, 1230, 1235, 1240, 1245, 1250, 1255, 1260, 1265, 1270, 1275, 1280, 1285, 1290, 1295, 1300, 1305, 1310, 1315, 1320, 1325, 1330, 1335, 1340, 1345, 1350, 1355, 1360, 1365, 1370, 1375, 1380, 1385, 1390, 1395, 1400, 1405, 1410, 1415, 1420, 1425, 1430, 1435, 1440, 1445, 1450, 1455, 1460, 1465, 1470, 1475, 1480, 1485, 1490, 1495, 1500, 1505, 1510, 1515, 1520, 1525, 1530, 1535, 1540, 1545, 1550, 1555, 1560, 1565, 1570, 1575, 1580, 1585, 1590, 1595, 1600, 1605, 1610, 1615, 1620, 1625, 1630, 1635, 1640, 1645, 1650, 1655, 1660, 1665, 1670, 1675, 1680, 1685, 1690, 1695, 1700, 1705, 1710, 1715, 1720, 1725, 1730, 1735, 1740, 1745, 1750, 1755, 1760, 1765, 1770, 1775, 1780, 1785, 1790, 1795, 1800, 1805, 1810, 1815, 1820, 1825, 1830, 1835, 1840, 1845, 1850, 1855, 1860, 1865, 1870, 1875, 1880, 1885, 1890, 1895, 1900, 1905, 1910, 1915, 1920, 1925, 1930, 1935, 1940, 1945, 1950, 1955, 1960, 1965, 1970, 1975, 1980, 1985, 1990, 1995, 2000, 2005, 2010, 2015, 2020, 2025, 2030, 2035, 2040, 2045, 2050, 2055, 2060, 2065, 2070, 2075, 2080, 2085, 2090, 2095, 2100, 2105, 2110, 2115, 2120, 2125, 2130, 2135, 2140, 2145, 2150, 2155, 2160, 2165, 2170, 2175, 2180, 2185, 2190, 2195, 2200, 2205, 2210, 2215, 2220, 2225, 2230, 2235, 2240, 2245, 2250, 2255, 2260, 2265, 2270, 2275, 2280, 2285, 2290, 2295, 2300, 2305, 2310, 2315, 2320, 2325, 2330, 2335, 2340, 2345, 2350, 2355, 2360, 2365, 2370, 2375, 2380, 2385, 2390, 2395, 2400, 2405, 2410, 2415, 2420, 2425, 2430, 2435, 2440, 2445, 2450, 2455, 2460, 2465, 2470, 2475, 2480, 2485, 2490, 2495, 2500, 2505, 2510, 2515, 2520, 2525, 2530, 2535, 2540, 2545, 2550, 2555, 2560, 2565, 2570, 2575, 2580, 2585, 2590, 2595, 2600, 2605, 2610, 2615, 2620, 2625, 2630, 2635, 2640, 2645, 2650, 2655, 2660, 2665, 2670, 2675, 2680, 2685, 2690, 2695, 2700, 2705, 2710, 2715, 2720, 2725, 2730, 2735, 2740, 2745, 2750, 2755, 2760, 2765, 2770, 2775, 2780, 27851190, 1195, 1200, 1205, 1210, 1215, 1220, 1225, 1230, 1235, 1240, 1245, 1250, 1255, 1260, 1265, 1270, 1275, 1280, 1285, 1290, 1295, 1300, 1305, 1310, 1315, 1320, 1325, 1330, 1335, 1340, 1345, 1350, 1355, 1360, 1365, 1370, 1375, 1380, 1385, 1390, 1395, 1400, 1405, 1410, 1415, 1420, 1425, 1430, 1435, 1440, 1445, 1450, 1455, 1460, 1465, 1470, 1475, 1480, 1485, 1490, 1495 mg / m 2 to about 1500 mg / m 2 ; from about 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 mg / m 2 to about 150 mg / m 2 ; or from about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 111, 11.5, 12, 12.5, 13, 13.5, 14, or 14.5 mg / m 2up to about 15 mg / m 2 .

[0220] VII. Dosage Form

[0221] A suitable pharmaceutical composition for use in the methods described herein can be formulated into a dosage form that can be administered to a patient. In one embodiment, the pharmaceutical composition is in the form of an oral dosage unit or a parenteral dosage unit. In one embodiment, the pharmaceutical composition is in the form of an oral dosage unit. In one embodiment, the oral dosage unit is divided into several smaller doses that are administered to the subject over a predetermined period of time to mitigate the toxicity of the therapeutic agent being administered. In one embodiment, the oral dosage unit is administered by a tablet or capsule that contains a controlled release formulation, which can comprise a number of particles, granules, pellets, microtablets, or tablets. In one embodiment, the pharmaceutical composition is in the form of a parenteral dosage unit. In one embodiment, the parenteral dosage unit is selected from the group consisting of intravenous (IV), subcutaneous (SC), intramuscular (M), rectal (PR), and transdermal dosage units. In one embodiment, the composition is in a dosage form selected from the group consisting of a sterile solution, a suspension, a suppository, a tablet, and a capsule. In one embodiment, the composition is an oral dosage form selected from the group consisting of a tablet, a caplet, a capsule, a lozenge, a syrup, a liquid, a suspension, and an elixir. In one embodiment, the composition is an oral dosage form selected from the group consisting of a tablet, a hard shell capsule, a soft gelatin capsule, a bead, a granule, an aggregate, a powder, a gel, a solid, and a semi-solid.

[0222] In one embodiment, a suitable pharmaceutical composition for use in the methods described herein is in the form of a dermatological composition suitable for topical administration to the skin. For example, the dermatological composition comprises a cosmetically or pharmaceutically acceptable carrier. The dermatological composition for topical administration can include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. In one embodiment, a conventional pharmaceutical carrier, aqueous, powder or oily base, thickening agents, skin enhancers can be necessary or desirable and thus be used. Examples of suitable enhancers include ethers such as diethylene glycol monoethyl ether (available as Transcutol® commercially available from Sigma-Aldrich®) and diethylene glycol monomethyl ether; surfactants such as sodium lauryl sulfate, sodium lauryl sulfate, cetyltrimethylammonium bromide, benzalkonium chloride, poloxamer (231, 182, 184), Tween (20, 40, 60, 80), and lecithin (U.S. Patent 4,783,450); alcohols such as ethanol, propanol, octanol, benzyl alcohol; polyethylene glycols and esters thereof such as polyethylene glycol monolaurate; amides and other nitrogen-containing compounds such as urea, dimethylacetamide (DMA), dimethylformamide (DMF), 2-pyrrolidone, 1-methyl-2-pyrrolidone, ethanolamine, diethanolamine, and triethanolamine; terpenes; alkanones; and organic acids, particularly citric acid and succinic acid. Also, it is possible to use commercially available from Sigma-Aldrich®) and diethylene glycol monomethyl ether; surfactants such as sodium lauryl sulfate, sodium lauryl sulfate, cetyltrimethylammonium bromide, benzalkonium chloride, poloxamer (231, 182, 184), Tween (20, 40, 60, 80), and lecithin (U.S. Patent 4,783,450); alcohols such as ethanol, propanol, octanol, benzyl alcohol; polyethylene glycols and esters thereof such as polyethylene glycol monolaurate; amides and other nitrogen-containing compounds such as urea, dimethylacetamide (DMA), dimethylformamide (DMF), 2-pyrrolidone, 1-methyl-2-pyrrolidone, ethanolamine, diethanolamine, and triethanolamine; terpenes; alkanones; and organic acids, particularly citric acid and succinic acid. Also, it is possible to use and sulfoxides such as DMSO and Cι0MSO, but are less preferred.

[0223] In one embodiment, the pharmaceutical composition is a dosage form selected from the group consisting of sustained release forms, controlled release forms, delayed release forms, and responsive release forms.

[0224] VIII. Methods of Use

[0225] The compositions and methods described herein have utility in treating many diseases, including cancers such as colorectal cancer, brain cancer, and glioblastoma. In one embodiment, the compositions and methods described herein are used to treat diseases such as ocular melanoma, desmoplastic round cell tumor, chondrosarcoma, leptomeningeal disease, diffuse large B-cell lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphoma, anal or rectal cancer, appendix cancer, astrocytoma, and atypical teratoid / rhabdoid tumor. In one embodiment, the compositions and methods described herein are used to treat diseases such as basal cell carcinoma, basal cell nevus syndrome, Gorlin nevus syndrome, bile duct cancer, bladder cancer, bone cancer, osteosarcoma and malignant fibrous histiocytoma, brain tumors, breast cancer, bronchial tumors, Burkitt's lymphoma, and spinal cord tumors. In one embodiment, the compositions and methods described herein are used to treat diseases such as carcinoid tumors, carcinoma of unknown primary, atypical teratoid / rhabdoid tumor of the central nervous system, leptomeningeal disease, embryonal tumor of the central nervous system, lymphoma of the central nervous system, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloproliferative disease, colon cancer, colorectal cancer, craniopharyngioma, and cutaneous T-cell lymphoma (including Sézary syndrome and mycosis fungoides (MF)). In one embodiment, the compositions and methods described herein are used to treat diseases such as embryonal tumor of the central nervous system, endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, Ewing's sarcoma family of tumors, extracranial germ cell tumors, extragonadal germ cell tumors, extrahepatic bile duct cancer, and eye cancers, including intraocular melanoma and retinoblastoma. In one embodiment, the compositions and methods described herein are used to treat diseases such as gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (GIST), germ cell tumors, gestational trophoblastic tumors, and gliomas. In one embodiment, the compositions and methods described herein are used to treat cancers selected from the group consisting of hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, histiocytosis, Hodgkin lymphoma, and hypopharyngeal cancer. In one embodiment, the compositions and methods described herein are used to treat diseases such as Kaposi Sarcoma and kidney (renal cell) cancer. In one embodiment, the compositions and methods described herein are used to treat diseases such as Langerhans cell histiocytosis, laryngeal cancer, lip and oral cancer, liver cancer, lung cancer, including non-small cell lung cancer and small cell lung cancer, non-Hodgkin lymphoma, and primary central nervous system lymphoma. In one embodiment, the compositions and methods described herein are used to treat diseases such as: Warren's disease, Macroglobulinemia (lymphoplasmacytic lymphoma), malignant fibrous histiocytoma and osteosarcoma of bone, medulloblastoma, medulloepithelioma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous neck cancer with occult primary, multiple endocrine neoplasia syndrome, oral cavity cancer, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myeloid leukemia with complex karyotype in blast phase, myeloproliferative disorder / myeloproliferative neoplasm, multiple myeloma, and myeloproliferative disease. In one embodiment, the compositions and methods described herein are used to treat cancer. In one embodiment, the compositions and methods described herein are used to treat diseases such as nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, and neuroblastoma. In one embodiment, the compositions and methods described herein are used to treat diseases such as oral cavity cancer, lip and oral cavity cancer, oropharyngeal cancer, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian cancer, ovarian germ cell tumor, ovarian epithelial cancer, and ovarian low malignant potential tumor. In one embodiment, the compositions and methods described herein are used to treat diseases such as pancreatic cancer, papilloma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pineal parenchymal tumors of intermediate differentiation, pineoblastoma and supratentorial primitive neuroectodermal tumors, pituitary tumor, pleuropulmonary blastoma, pregnancy and breast cancer, primary central nervous system lymphoma, and prostate cancer. In one embodiment, the compositions and methods described herein are used to treat a cancer selected from the group consisting of rectal cancer, renal cell (kidney) cancer, renal pelvis and ureter cancer, respiratory tract carcinoma involving the NUT gene on chromosome 15, retinoblastoma, and rhabdomyosarcoma. In one embodiment, the compositions and methods described herein are used to treat high-grade prostate cancer. In one embodiment, the compositions and methods described herein are used to treat intermediate-grade prostate cancer. In one embodiment, the compositions and methods described herein are used to treat low-grade prostate cancer. In one embodiment, the compositions and methods described herein are used to treat castration-resistant prostate cancer. In one embodiment, the compositions and methods described herein are used to treat a nervous system tumor. In one embodiment, the compositions and methods described herein are used to treat a central nervous system tumor. In one embodiment, the compositions and methods described herein are used to treat a peripheral nervous system tumor. In one embodiment, the compositions and methods described herein are used to treat a paraganglioma. In one embodiment, the compositions and methods described herein are used to treat a pheochromocytoma.

[0226] Compound (1) (ONC201) has broad anti-cancer activity, low toxicity, including few adverse effects if any, low genotoxicity, and high bioavailability, including oral bioavailability, in in vitro models, animal models, and human clinical trials. These characteristics make ONC201 and various analogs particularly well suited for pediatric patients. These characteristics also make ONC201 and various analogs particularly well suited for long-term treatment, high-risk patients, and to maintain a long response or stable disease or prevent disease recurrence.

[0227] In one embodiment, the compositions and methods described herein are used to treat pediatric cancers (e.g., pediatric solid tumors, pediatric sarcomas, pediatric Ewing's sarcoma, pediatric gliomas, pediatric central nervous system cancers, pediatric neuroblastomas, pediatric leukemias, and pediatric lymphomas).

[0228] In one embodiment, the compositions and methods described herein are used to treat proliferative skin diseases, such as psoriasis. In one embodiment, the compositions and methods described herein are used to treat a cancer selected from the group consisting of salivary gland carcinoma, sarcoma, Sezary syndrome, skin cancer, ocular cancer, cutaneous carcinoma, small bowel cancer, soft tissue sarcoma, squamous cell carcinoma, squamous cell carcinoma of the neck with occult primary, and supratentorial primitive neuroectodermal tumor. In one embodiment, the compositions and methods described herein are used to treat a cancer selected from the group consisting of T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell cancer of the renal pelvis and ureter, and trophoblastic tumor of pregnancy. In one embodiment, the compositions and methods described herein are used to treat a cancer selected from the group consisting of carcinoma of unknown primary site, childhood abnormal cancer, transitional cell cancer of the renal pelvis and ureter, urethral cancer, and uterine sarcoma. In one embodiment, the compositions and methods described herein are used to treat a cancer selected from the group consisting of vaginal cancer and vulvar cancer. In one embodiment, the compositions and methods described herein are used to treat a cancer selected from the group consisting of Wilms' tumor and female cancer.

[0229] In one embodiment, the compositions and methods described herein are used as a first line therapy (sometimes referred to as a primary therapy). In one embodiment, the compositions and methods described herein are used as a second line therapy. In one embodiment, the compositions and methods described herein are used as a third line therapy. In one embodiment, the compositions and methods described herein are used as a salvage therapy. The term "salvage therapy" refers to a therapeutic agent that can be employed in any regimen after a subject's initial treatment regimen has failed or after a subject's condition has not responded to the initial treatment. In one embodiment, the compositions and methods described herein are used as a salvage therapy. In one embodiment of a salvage therapy, the compositions are used as a salvage agent to counteract the effects of an initial treatment. In one embodiment of a salvage therapy, the compositions are used as a salvage agent administered to a subject who has become resistant to a standard or initial treatment. In one embodiment, the compositions and methods described herein are used as a neoadjuvant therapy. In one embodiment, a neoadjuvant therapy comprises administering one or more therapeutic agents described herein to a subject prior to a primary or first line therapy. In one embodiment, the neoadjuvant therapy reduces the size or extent of a cancer being treated prior to administering a primary or first line therapy to a subject being treated. In one embodiment, the compositions and methods described herein are used as an adjuvant therapy. In one embodiment, an adjuvant therapy comprises administering one or more therapeutic agents described herein to a subject, wherein the one or more therapeutic agents alter the effects of other therapeutic agents that have been administered to the subject or are administered to the subject concurrently or subsequently.

[0230] In one embodiment, the compositions and methods described herein exhibit a reduced probability of drug-drug interaction. In one embodiment, eunotin, e.g., ONC201, or an analog thereof, is eliminated from the patient's body before it can interact with another pharmaceutically active agent.

[0231] In one embodiment, the compositions and methods described herein exhibit a toxicity level that facilitates combination with other pharmaceutical agents.

[0232] The methods and compositions described herein are not limited to a particular animal species. In one embodiment, a subject treated in accordance with the methods described herein and using the compositions described herein can be a mammal or a non-mammal. In one embodiment, a mammal subject mammal includes, but is not limited to, a human; a non-human primate; a rodent such as a mouse, rat, or guinea pig; a domestic pet such as a cat or dog; a horse, cow, pig, sheep, goat, or rabbit. In one embodiment, a non-mammal subject includes, but is not limited to, a bird such as a duck, goose, chicken, or turkey. In one embodiment, the subject is a human. In one embodiment, the subject can be of either sex and of any age. The compositions and methods can also be used to prevent cancer. The compositions and methods can also be used to stimulate the immune system.

[0233] The methods and compositions described herein are not limited to a particular age of the subject. In one embodiment, the subject treated according to the methods described herein and using the compositions described herein is over the age of 50 years, over the age of 55 years, over the age of 60 years, or over the age of 65 years. In one embodiment, the subject treated according to the methods described herein and using the compositions described herein is under the age of 50 years, under the age of 55 years, under the age of 60 years, or under the age of 65 years.

[0234] In one embodiment, the subject treated according to the methods described herein and using the compositions described herein is a pediatric patient. In one embodiment, the pediatric patient is less than 18 years of age, less than 17 years of age, less than 16 years of age, less than 15 years of age, less than 14 years of age, less than 13 years of age, less than 12 years of age, less than 11 years of age, less than 10 years of age, less than 9 years of age, less than 8 years of age, less than 7 years of age, less than 6 years of age, less than 5 years of age, less than 4 years of age, less than 3 years of age, less than 2 years of age, or less than 1 year of age. In one embodiment, the pediatric patient is less than 12 months of age, less than 11 months of age, less than 10 months of age, less than 9 months of age, less than 8 months of age, less than 7 months of age, less than 6 months of age, less than 5 months of age, less than 4 months of age, less than 3 months of age, less than 2 months of age, or less than 1 month of age. In one embodiment, the pediatric patient is less than 4 weeks of age, less than 3 weeks of age, less than 2 weeks of age, or less than 1 week of age. In one embodiment, the pediatric patient is less than 7 days of age, less than 6 days of age, less than 5 days of age, less than 4 days of age, less than 3 days of age, less than 2 days of age, or less than 1 day of age. In one embodiment, the pediatric patient is a neonate. In one embodiment, the pediatric patient is a premature infant.

[0235] In one embodiment, the patient weighs less than 45 kg, less than 40 kg, less than 35 kg, less than 30 kg, less than 25 kg, less than 20 kg, less than 15 kg, less than 14 kg, less than 10 kg, less than 5 kg, less than 4 kg, less than 3 kg, less than 2 kg, or less than 1 kg.

[0236] In one embodiment, the subject has received at least one prior therapeutic agent. In one embodiment, the subject has received at least two, at least three, or at least four prior therapeutic agents. In one embodiment, the prior therapeutic agent is ibrutinib, bortezomib, carfilzomib, temozolomide, bevacizumab, cyclophosphamide, hydroxydaunorubicin, vincristine, prednisone, cytarabine, cisplatin, rituximab, 5-fluorouracil, oxaliplatin, leucovorin, or lenalidomide.

[0237] In one embodiment, the subject has been treated with radiation therapy. In one embodiment, the subject has been treated with surgery. In one embodiment, the subject has been treated with adoptive T cell therapy.

[0238] In one embodiment, the cancer is no longer responsive to treatment with ibrutinib, bortezomib, carfilzomib, temozolomide, bevacizumab, cyclophosphamide, hydroxydaunomycin, vincristine, prednisone, cytarabine, cisplatin, rituximab, 5-fluorouracil, oxaliplatin, folinic acid, lenalidomide, radiation, surgery, or a combination thereof.

[0239] In one embodiment, the dose response relationship of the compositions and methods described herein in cancer cells is different from the dose response relationship of the same compositions and methods in normal cells. The dose response relationship of ONC201 to proliferation and cell death in normal cells and tumor cells was determined by measuring cell viability after 72 hours of treatment with various concentrations of ONC201. The tumors tested included human colon carcinoma cell line (HCT116), breast tumor cell line (MDA-MB-231), and human primary glioblastoma cell line (U87). And the normal cells tested included human foreskin fibroblast (HFF), human fetal lung fibroblast (MRC-5) cells, and human lung fibroblast line (WI-38). Doxorubicin was used as a positive control in normal fibroblasts at 1 pg / mL. The cell viability of the normal cells tested was at least about 75% at about 1-5 mg / mL ONC201, while the viability of the tumor cells was significantly lower (e.g., equal to or less than 50%) at the same ONC201 concentration. Further, as the concentration of ONC201 increased beyond about 5 mg / mL, the viability of the tumor cells decreased to less than 25%, while the viability of the normal cells remained at about 75%. Cell viability assays in human fetal lung fibroblast (MRC-5) cells were performed after 72 hours of treatment with Compound (1) (5 mM) or DMSO and after a recovery period in complete drug-free medium after treatment. Cell recovery was seen with ONC201, but not with DMSO.

[0240] In one embodiment, the compositions and methods described herein are useful in treating cancer in a subject. In one embodiment, the compositions and methods described herein are useful in treating cancer in a human subject. In one embodiment, the method of treatment comprises administering to a subject in need of such treatment a pharmaceutically effective amount of imirichmane, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0241] In one embodiment, the method of treatment comprises administering to a subject in need of such treatment: (i) a first therapeutic agent comprising imirichne, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in combination with (ii) a second therapeutic agent, wherein the first and second therapeutic agents are administered simultaneously or sequentially. The second therapeutic agent can be any suitable therapeutic agent, including any of the pharmaceutically active agents disclosed herein. Pharmaceutically acceptable salts of ONC201 include the following dihydrochloride salt:

[0242]

[0243] It is understood that the dihydrochloride salt of ONC201 or an analog thereof, including the compound of formula (10), or an alternative dihydrochloride salt thereof apparent from the teachings of the present disclosure, can be substituted for ONC201 or an analog thereof in the compositions or administration regimens described herein.

[0244] In one embodiment, the method of treatment comprises simultaneously or sequentially administering to a subject in need of such treatment a synergistic drug combination, wherein the synergistic drug combination comprises: (i) a first therapeutic agent comprising imirichne, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) a second therapeutic agent. In one embodiment, the method of treatment comprises simultaneously or sequentially administering to a subject in need of such treatment a therapeutically synergistic amount of a combination of a first therapeutic agent and a second therapeutic agent. In one embodiment, the method of treatment comprises simultaneously or sequentially administering to a subject in need of such treatment an effective amount of a first therapeutic agent in combination with an effective amount of a second therapeutic agent, wherein the combination provides a synergistic effect in treating in vivo a cancer susceptible to the combination, and wherein the first and second therapeutic agents are administered simultaneously or sequentially. In one embodiment, the method of treatment comprises administering to a subject in need of such treatment an effective amount of a first therapeutic agent in combination with an effective amount of a second therapeutic agent, wherein the combination provides a synergistic effect in treating in vivo minimal residual disease susceptible to the combination, and wherein the first and second therapeutic agents are administered simultaneously or sequentially. In one embodiment, the second therapeutic agent is administered prior to or before the first therapeutic agent.

[0245] In one embodiment, the method of treatment targets a cancer selected from the group consisting of a solid tumor, a liquid tumor, a lymphoma, a leukemia, or a myeloma.

[0246] In one embodiment, the method of treatment targets a solid tumor, wherein the solid tumor is selected from the group consisting of: uterine cervical cancer, endometrial cancer, extracranial germ cell tumor; extragonadal germ cell tumor; gestational trophoblastic tumor; ovarian cancer, ovarian germ cell tumor, ovarian epithelial cancer, and ovarian malignant malignant tumor; penile cancer, prostate cancer; pregnancy and breast cancer high grade prostate cancer; carrier grade prostate cancer; low grade prostate cancer; castration resistant prostate cancer; breast cancer; cholangiocarcinoma; extrahepatic bile duct cancer; gallbladder cancer hepatocellular carcinoma (liver cancer) kidney (renal cell) cancer; liver cancer, renal cell (kidney) cancer, renal pelvis and ureter; basal cell carcinoma; basal cell nevus syndrome, Gorlin-Nevus syndrome, melanoma, Merkel cell carcinoma, papillomatosis, multiple endocrine neoplasia syndrome; pancreatic cancer, parathyroid cancer, ocular melanoma; eye cancer; retinoblastoma; malignant fibrous histiocytoma Ewing sarcoma family tumor; desmoplastic small round cell tumor; chondrosarcoma, Kaposi sarcoma, rhabdomyosarcoma; spinal cord tumor, meningitis, central nervous system embryonic tumor, chordoma, central nervous system embryonic tumor, endothelioma, ependymoma, neuroblastoma; intermediate-differentiated pineal parenchymal tumor, pineal cell tumor; adrenocortical carcinoma; bone cancer, osteosarcoma bone and malignant fibrous histiocytoma; bone sarcoma and malignant fibrous histiocytoma bone; carcinoid tumor, cancer of unknown primary, bronchial tumor, lung cancer, pleuropulmonary blastoma; NUT gene- involved 15-chromosome respiratory tract cancer, astrocytoma, atypical teratoid / rhabdoid tumor; central nervous system atypical teratoid / rhabdoid tumor, craniopharyngioma, glioma, brain cancer, medulloblastoma, medullary epithelioma, epithelial primitive neuroectodermal tumor; pituitary tumor stomach cancer (stomach), gastrointestinal carcinoid, gastrointestinal stromal tumor (GIST), bladder cancer, anorectal cancer, adnexal cancer, esophageal cancer, hypopharyngeal cancer; laryngeal cancer, lip and oral cavity cancer, metastatic squamous cell carcinoma with occult primary, oral cavity cancer, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, oral cavity and oral cavity cancer, oropharyngeal cancer, paranasal sinus and nasal cavity cancer, pharyngeal cancer; head and neck cancer and mesothelioma.

[0247] In one embodiment, the method of treatment targets a lymphoma selected from the group consisting of: diffuse large cell lymphoma, AIDS-related lymphoma, cutaneous T-cell lymphoma, Sezary syndrome, mycosis fungoides (MF); histiocytosis; Burkitt lymphoma and central nervous system lymphoma; non-Hodgkin lymphoma, primary central nervous system lymphoma, Hodgkin lymphoma, macroglobulinemia; mycosis fungoides primary central nervous system lymphoma; lymphoplasmacytic lymphoma and primary central nervous system lymphoma.

[0248] In one embodiment, the method of treatment targets a non-Hodgkin lymphoma (NHL) selected from the group consisting of: mantle cell lymphoma, diffuse large cell lymphoma, follicular lymphoma, marginal zone lymphoma, small lymphocytic lymphoma, lymphocytic lymphoma NHL, Waldenstrom macroglobulinemia, and cutaneous lymphoma.

[0249] In one embodiment, the method of treatment targets a leukemia selected from the group consisting of: acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloproliferative disorders; hairy cell leukemia acute myeloid leukemia (AML); chronic myeloid leukemia (CML); and Langerhans cell histiocytosis.

[0250] In one embodiment, the method of treatment targets an acute leukemia selected from the group consisting of: acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, myelodysplastic syndrome, and myeloproliferative disorder.

[0251] In one embodiment, the method of treatment targets a myeloma selected from the group consisting of: IgA myeloma; IgG myeloma; IgM myeloma; IgD myeloma; IgE myeloma; light chain myeloma; non-secretory myeloma; complex karyotype, blastic phase leukemia; multiple myeloma / plasma cell neoplasm, multiple myeloma, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, and myeloproliferative disorder.

[0252] In one embodiment, the method of treatment targets a peripheral nervous system tumor. In one embodiment, the method of treatment targets a paraganglioma. In one embodiment, the method of treatment targets a pheochromocytoma.

[0253] In one embodiment, the treatment of cancer comprises preventing tumor growth in a cancer subject. In one embodiment, the treatment of cancer comprises preventing metastasis formation in a cancer subject. In one embodiment, the treatment of cancer comprises targeting minimal residual disease in a cancer subject known to have or at risk of having minimal residual disease in a cancer subject.

[0254] This can be indicated after treatment of the primary tumor by surgery and / or after chemotherapy (radiotherapy) has been initiated or determined to be effective. Disseminated tumor cells can be in their dormant state and often cannot be attacked by chemotherapy (radiotherapy). Patients treated this way appear to be in a cured state and are referred to as "minimal residual disease". However, if the dormant tumor cells become metastatic cells after a longer dormant state due to a growth stimulus, they have the potential to form metastases.

[0255] The term "minimal residual disease" refers to the small number of cancer cells that remain in a subject during or after treatment when the subject is in remission (does not exhibit symptoms or signs of disease). The methods described herein are preferably applied to the disease forms listed herein, including adult and pediatric forms of these diseases.

[0256] In one embodiment, the method of treatment can be used to treat autoimmune diseases. Autoimmune diseases include, but are not limited to, alopecia areata, antiphospholipid, autoimmune hepatitis, celiac disease, diabetes mellitus type 1, Graves' disease, Guillain-Barre syndrome, Hashimoto's disease, hemolytic anemia, idiopathic thrombocytopenic purpura, inflammatory bowel disease, inflammatory myopathy, multiple sclerosis, primary biliary cirrhosis, psoriasis, rheumatoid arthritis, scleroderma, syndrome, systemic lupus erythematosus, and vitiligo.

[0257] In one embodiment, the method of treatment can be used to treat autoimmune and inflammatory diseases of the peripheral nervous system such as amyotrophic lateral sclerosis (Lou Gehrig's disease) based on various causes, for example, metabolic disorders including diabetes, B12 and folate vitamin deficiencies, chemotherapy drugs and medications for treatment of HIV, poisons that cause peripheral nerve damage, cancers that develop peripheral neuropathy, paraneoplastic syndromes, alcohol abuse, chronic kidney disease, injuries that cause compression of nerves and other lesions such as Lyme disease, Guillain-Barre syndrome, connective tissue diseases, rheumatoid arthritis, Sjogren's syndrome, systemic lupus erythematosus, certain inflammatory conditions such as sarcoidosis, celiac disease, inherited diseases such as charcot marie tooth syndrome, Friedreich's ataxia, and / or idiopathic where no specific cause is found but inflammation and / or autoimmune mechanisms are believed to be the cause of the disease.

[0258] In one embodiment, the method of treatment can be used to treat autoimmune and inflammatory diseases with ocular manifestations. Such ocular manifestations include, but are not limited to, ocular cicatricial pemphigoid, Mooren's ulcer, various forms of uveitis, rheumatoid arthritis, systemic lupus erythematosus, polyarteritis nodosa, relapsing polychondritis, Wegener's granulomatosis, scleroderma, Behcet's disease, Reiter's disease, inflammatory bowel disease (ulcerative colitis and Crohn's disease), and ankylosing spondylitis, retinitis pigmentosa, macular degeneration, keratoconjunctivitis, scleritis, episcleritis, keratitis, peripheral corneal ulceration, and less commonly entities such as choroiditis, retinal vasculitis, scleral nodules, retinal detachment, and / or macular edema.

[0259] In one embodiment, the method of treatment can be used to treat acute allograft rejection in transplant patients. In one embodiment, the method of treatment can be used to treat ischemic stroke. In one embodiment, the method of treatment is useful for treating inflammatory diseases, including arthritis, psoriasis, asthma, and colitis.

[0260] In one embodiment, the therapeutic agent comprises a pharmaceutically acceptable monosalt of ONC201 or an analog thereof (e.g., a compound of formula (10)). In one embodiment, the therapeutic agent comprises a pharmaceutically acceptable disalt of ONC201 or an analog thereof (e.g., a compound of formula (10)). As described herein, some analogs may be trisalts. In one embodiment, the therapeutic agent comprises a monosalt or disalt of ONC201 or an analog thereof (e.g., a compound of formula (10)) in a pharmaceutically acceptable form selected from the group consisting of hydrochloride, hydrobromide, bisulfate, sulfate, phosphate, fumarate, succinate, oxalate, and lactate, bisulfate, hydroxy, tartrate, nitrate, citrate, tartrate, carbonate, malate, maleate, fumarate, sulfonate, methanesulfonate, formate, acetate, and carboxyl. In one embodiment, the therapeutic agent comprises ONC201 or an analog thereof in the form of a pharmaceutically acceptable salt selected from the group consisting of toluenesulfonate, benzenesulfonate, methanesulfonate, oxalate, succinate, tartrate, citrate, fumarate, and maleate. In one embodiment, the therapeutic agent comprises ONC201 or an analog thereof in the form of a pharmaceutically acceptable mono- or di-salt with a counterion selected from the group consisting of ammonium, sodium, potassium, calcium, magnesium, zinc, lithium, and / or with a counterion such as methylamino, dimethylamino, diethylamino, triethylamino, and combinations thereof. In one embodiment, the therapeutic agent comprises a di-salt of a compound described herein in the form of a halide, such as a dihydrochloride or dihydrobromide salt.

[0261] In one embodiment of the method of treatment, the second therapeutic agent comprises an anticancer agent. In one embodiment of the method of treatment, the second therapeutic agent is selected from abamectin, aclarubicin, atradazole, axonin, aspirin, alderleukin, ivermectin, allopurinol, atrazine, cyclomycin, isoproterenol, amifostine, aminoglutarimide, anastrozole, anseramicin, arsenic trioxide, asparaginase, astrin, azacitidine, azetepa, azotomycin, batimastat, benzoglucose, bevacizumab, bicalutamide, bisantrene, binefadol dimesylate, bieneuxin, bleomycin, brequinar, bropiridone, busulfan, cetirizine, capecitabine, capecitabine, carboplatin, carmustine, carbenicillin ... Carfilzomib, celecoxib, chlorambucil, siromulin, cisplatin, cladribine, cinorbendazole mesylate, cyclophosphamide, cytarabine, dacarbazine, dactinomycin D, daunorubicin, decitabine, dexamethasone, deazaguanine, dezagunium mesylate, diazolin, docetaxel, doxorubicin, droloxifene, dibromomorphone, duroxamine, edatrexamine, etisulin, edatrexamine, cisplatin, etoposide, etopostatin, etizole, etoposide, etopostatin, fadazole, famustine, fenretinide, fluorouracil, fludarabine, fluorouracil, flucitabine, fluoxetine, forsterol, fulvestrant, gemcitabine, hydroxyurea, daunorubicin, ifosfamide, sarcosine, interleukin II (IL-2 2, including recombinant interleukin II or rIL2), interferon alfa-2a, interferon alfa-2b, interferon alfa-1, interferon alfa-3, interferon beta-1a, midazolam, midazolam, meropenem, mecamyl, mecamyl, mercaptopurine, methotrexate, metropine, meturedepa, mimodex, mitropine, metroxanthine, mitomycin, mitomycin, mitomycin, mitomycin, mitosper, mitotane, mitoxantrone, mycophenolic acid, nelarabine, nocodazole, nokaramycin, olanzapine, oxiconazole, paclitaxel, pegaspargase, promoxetine, promoxetine, pipobroman, piposulfa n, piroxatone hydrochloride, mithramycin, plomimane, porfimer, porfimromycin, prednimustine, procarbazine, puromycin, pyrrolidone, riboprine, rogletimide, safingol, semustine, simtrazene, sparsosycin, sparsomycin, spirogermanium, spiromustine, streptozotocin, sulofenur, tebimycin, tamoxifen, teigatran, tegafur, tetostatin, temoporfin, teniposide, teroxirone, testolactone,Thiamiprine, thioguanine, thiotepa, thiazofuran, telapazolamide, topotecan, toremifene, trestolone, vitamin B, vitamin B, verinolactone, vitamin B, vibrinol, vezoloxetine, vezolanol, zenaplatin, zonotin, zoledronic acid, zolimus, and combinations thereof. ,

[0262] In some embodiments of the method of treatment, the second therapeutic agent is selected from the group consisting of hormone analogs and anti-hormones, aromatase inhibitors, LHRH agonists and antagonists, growth factor inhibitors, growth factor antibodies, growth factor receptor antibodies, tyrosine kinase inhibitors; antimetabolites; antitumor antibiotics; platinum derivatives; alkylating agents; antimitotic agents S; tubulin inhibitors; PARP inhibitors, topoisomerase inhibitors, serine / threonine kinase inhibitors, tyrosine kinase inhibitors, protein-protein interaction inhibitors, MEK inhibitors, ERK inhibitors, IGF-1R inhibitors, ErbB receptor inhibitors, rapamycin analogs, amifostine, anagrelide, clodronate, filgrastim, interferon, interferon alpha, folinic acid, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate and porfimer, amifostine, anagrelide, clodronate, filgrastim, interferon, interferon alpha, folinic acid, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate and porfimer, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent is selected from the group consisting of 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-alethine, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxy camptothecin, 16-aza-epothilone B, A 105972, A204197, abiraterone, aldesleukin, acitretin, allovectin-7, altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037, AP-5280, apaziquone, apomine, aranose, arglabin, arzoxifene, atamestane, atrasentan, auristatin PE, ABT-199 (Venetoclax), ABT-263 (Navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), ARRY 162, ARRY 438162, ARRY-300, ARRY-142886 / AZD-6244 (selumetinib), ARRY-704 / AZD-8330, AR-12, AR-42, AS-703988, AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ680, AS-703026 (primasertib), avastin, AZD-2014, azacitidine, ixabepilone B, azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, binimetinib dichloride, BCX-1777, BKM-120, bliconin, BLP-25, BMS-184476,BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW 2992 (afatinib, Gilotrif), BIBF 1120 (vargagef), BI 836845, BI 2536, BI 6727, BI 836845, BI 847325, BI 853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryostatins-1, bortezomib, brostallicin, busulfan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, canertinib, canfosfamide, capecitabine, carboplatin, CCI-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, ceflatonin, ceftriaxone, celecoxib, celmoleukin, cemadotin, CH4987655 / RO-4987655, chloranil, ceilidin, cyclosporin, CDA-II, CKD-394, CKD-602, CKI-27, clofarabine, colchicine, combretastin A4, COT inhibitor, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTL A-4 monoclonal antibody, CP-461, CV-247, cyanomorpholino doxorubicin, cytarabine, D24851, decitabine, doxorubicin, deoxyrubicin, deoxycoformycin, depsipeptide, deoxyepothilone B, dexamethasone, dexrazoxane, dienestrol, diflomotecan, didox, DMDC, dolastatin 10, dolastatins, DS-7423, E7010, E-6201, edatrexat, edotreotide, efaproxiral, efomycin, EGFR inhibitor, EKB-569, EKB-509, enzastaurin, enzalomid, enoxacin, epothilone B, Epratuzumab, ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine, ethynyloestradiol, exatecan, exatecan mesylate, exemestane, exisulind, fenretinide, etomoxir, floxuridine, folic acid, FOLFOX, FOLFOX4, FOLFIRI, fostedil, flutaxim, gallium maltolate, gefitinib, gemtuzumab, gemcitabine, GLC-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen,GMK, GPX-100, gp100 peptide vaccine, GSK-5126766, GSK-690693, GSK-1120212 (trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron, herceptin, hexamethylmelamine, histamine, homoharringtonine, hyaluronic acid, hydroxyurea, hydroxypregesterone caproate, ibandronate, ibritumomab tiuxetan, idatrexate, idenestrol, IDN-5109, IGF-1R inhibitor, IMC-1C11, IMC-A12 (cixutumumab), immunol, indisulam, interferon alpha-2a, interferon alpha-2b, pegylated interferon alpha-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib, ipilimumab, iproplatin, irafon, isohomohalichondrin-B, isoflavones, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, kahalid F, ketoconazole, KW-2170, KW-2450, lobaplatin, leflunomide, lenograstim, leuprolide, lexidronam, LGD-1550, linezolid, lutathera, lomustine, losoxantrone, LU 223651, losoxantrone, LY-S6 AKT1, LY-2780301, mafosfamide, marimastat, mechlorethamine, MEK inhibitor, MEK-162, methyltestosterone, methylprednisolone, MEDI-573, MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, midostaurin, minodronic acid, mitomycin, mivobulin, MK-2206, MK-0646 (dalotuzumab), MLN518, motexafin gadolinium, MS-209, MS-275, MX6, neridronic acid, neratinib, Nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nolatrexed, norlestrin, N-acetylcysteine, 06-benzylguanine, Oblimersen, omeprazole, oncophage, oncoVEX GM-CSF, ormiplatin, ortataxel, OX44 antibody, OSI-027, OSI-906 (linsitinib), 4-1BB antibody, oxantrazole, estrogen, panitumumab, patupilone, pegfilgrastim, PCK-3145, pegfilgrastim, PBI-1402, PBI-05204, PDO325901,PD-1 antibody, PEG-paclitaxel, albumin-stabilized paclitaxel, PEP-005, PF-05197281, PF-05212384, PF-04691502, PHT-427, P-04, PKC412, P54, PI-88, pelitinib, premetrexed, pentrix, pirfenidone, plicatinol, pertuzumab, PI3K inhibitor, PI3K / mTOR inhibitor, PG-TXL, PG2, PLX-4032 / RO-5185426 (vemurafenib), PLX-3603 / RO-5212054, PT-100, PWT-33597, PX-866, platinol, pivaloyloxymethyl butyrate, primidone, phenytoin O, PKI166, platinol, plicamycin, polyprenic acid, porfiromycin, prednisone, prednisolone, quinamed, quinupristin, R115777, RAF-265, ramucirumab, ranpirnase, RDEA-119 / BAY 869766, RDEA-436, rebecamycin analog, receptor tyrosine kinase (RTK) inhibitor, revimid, RG-7167, RG-7304, RG-7421, RG-7321, RG 7440, rhizoxin, rhu-MAb, rilpivirine, risedronate, rituximab, rofecoxib, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR 109881A, rubidomycin phenylhydrazone, rubitecan, R-flurbiprofen, RX-0201, S-9788, sabarubicin, SAHA, sargramostim, satraplatin, SB 408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustin, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine, suberanilohydroxamic acid, sutent, T 900607, T 138067, TAK-733, TAS-103, tacedinaline, talabostat, tariquitar, tasisulam, taxotere, taxoprexin, tazarotene, tegafur, temozolomide, temilifene, testosterone, testosterone propionate, temilifene, tetraplatin, tetrodotoxin, tezacitabine, thalidomide, theralux, therarubicin, thymalfasin, thymectacin, tiazofurin, tipifarnib, tirapazamine, tozasamine, trademere, toremofin, triciribine, trans-MID-107,Trans-retinoic acid, traszutumab, tremelimumab, tretinoin, triacetyluridine, triapine, triciribine, trimetrexate, TLK-286TXD 258, tykerb / tyverb, urocidin, valrubicin, vatalanib, vincristine, vinflunine, virulizine, WX-UK1, WX-554, vectibix, xeloda, XELOX, XL-147, XL-228, XL-281, XL-518 / R-7420 / GDC-0973, XL-765, YM-511, YM-598, ZD-4190, ZD-6474, ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZD1839, ZSTK-474, zoledronat, zosuquidar, and combinations thereof.

[0263] In one embodiment of the method of treatment, the second therapeutic agent is selected from tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate, flutamide, nilutamide, bicalutamide, aminoglutethimide, cyproterone acetate, finasteride, buserelin acetate, fludrocortisone, fluoxymesterone, medroxyprogesterone, octreotide, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent is selected from LHRH agonists and LHRH antagonists. In one embodiment, the LHRH agonist is selected from goserelin acetate, loprilide acetate, triptorelin pamoate, and combinations thereof. In one embodiment, the second therapeutic agent comprises an LHRH antagonist selected from degarelix, cetrorelix, abarelix, ozarelix, degarelix, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises an inhibitor of a growth factor. In one embodiment, the inhibitor of the growth factor is selected from the group consisting of platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), insulin-like growth factor (IGF), human epidermal growth factor (HER), hepatocyte growth factor (HGF), and combinations thereof. In one embodiment, the human epidermal growth factor (HER) is selected from HER2, HER3, and HER4.

[0264] In one embodiment of this treatment method, the second therapeutic agent comprises a tyrosine kinase inhibitor. In one embodiment of this treatment method, the tyrosine kinase inhibitor is selected from cetuximab, gefitinib, imatinib, lapatinib, and trastuzumab, and combinations thereof. In one embodiment of this treatment method, the second therapeutic agent comprises an aromatase inhibitor. In one embodiment of this treatment method, the aromatase inhibitor is selected from anastrozole, letrozole, rirozole, vorozole, exemestane, atamestane, and combinations thereof.

[0265] In one embodiment of the treatment method, the second therapeutic agent comprises an antimetabolite. In one embodiment of the treatment method, the antimetabolite comprises an antifolate. In one embodiment of the treatment method, the antifolate is selected from the group consisting of methotrexate, raltitrexed, pyrimidine analogs, and combinations thereof. In one embodiment of the treatment method, the antimetabolite is a pyrimidine analog. In one embodiment of the treatment method, the pyrimidine analog is selected from the group consisting of 5-fluorouracil, capecitabine, gemcitabine, and combinations thereof. In one embodiment of the treatment method, the antimetabolite is a purine analog or an adenosine analog. In one embodiment of the treatment method, the purine analog or adenosine analog is selected from the group consisting of mercaptopurine, thioguanine, cladribine, and pentostatin, cytarabine, fludarabine, and combinations thereof. In one embodiment of the treatment method, the second therapeutic agent comprises an antitumor antibiotic. In one embodiment of the treatment method, the antitumor antibiotic is selected from the group consisting of anthracyclines, doxorubicin, daunorubicin, epirubicin and idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin, streptozocin, and combinations thereof. In one embodiment of the treatment method, the second therapeutic agent comprises a platinum derivative. In one embodiment of the treatment method, the platinum derivative is selected from the group consisting of cisplatin, oxaliplatin, carboplatin, and combinations thereof. In one embodiment of the treatment method, the second therapeutic agent comprises an alkylating agent. In one embodiment of the treatment method, the alkylating agent is selected from the group consisting of estramustine, mechlorethamine, melphalan, chlorambucil, busulfan, dacarbazine, cyclophosphamide, ifosfamide, temozolomide, nitrosoureas, and combinations thereof. In one embodiment of the treatment method, the second therapeutic agent comprises a nitrosourea. In one embodiment of the treatment method, the nitrosourea is selected from the group consisting of carmustine, lomustine, thiotepa, and combinations thereof. In one embodiment of the treatment method, the second therapeutic agent comprises an antimitotic agent. In one embodiment of the treatment method, the antimitotic agent is selected from the group consisting of vinca alkaloids and taxanes. In one embodiment of the treatment method, the taxane is selected from the group consisting of paclitaxel, docetaxel, and combinations thereof. In one embodiment of the treatment method, the vinca alkaloid is selected from the group consisting of vinblastine, vindesine, vinorelbine, vincristine, and combinations thereof. In one embodiment of the treatment method, the second therapeutic agent comprises a topoisomerase inhibitor. In one embodiment of the treatment method, the topoisomerase inhibitor is an epipodophyllotoxin. In one embodiment of the treatment method, the topoisomerase inhibitor is an epipodophyllotoxin selected from the group consisting of etoposide, teniposide, nimustine, amsacrin, topotecan, irinotecan, mitoxantrone, and combinations thereof. In one embodiment of the treatment method, the second therapeutic agent comprises a serine / threonine kinase inhibitor. In one embodiment of the treatment method, the serine / threonine kinase inhibitor is selected from the group consisting of PDK1 inhibitors, B-Raf inhibitors, mTOR inhibitors, mTORCl inhibitors, PI3K inhibitors, dual mTOR / PI3K inhibitors, STK 33 inhibitors, AKT inhibitors, PLK 1 inhibitors, CDK inhibitors, Aurora kinase inhibitors, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises a tyrosine kinase inhibitor. In one embodiment of the method of treatment, the second therapeutic agent comprises a PTK2 / FAK inhibitor. In one embodiment of the method of treatment, the second therapeutic agent comprises a protein-protein interaction inhibitor. In one embodiment of the method of treatment, the protein-protein interaction inhibitor is selected from the group consisting of IAP, Mcl-1, MDM2 / MDMX, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises a rapamycin analog. In one embodiment of the method of treatment, the rapamycin analog is selected from the group consisting of everolimus, temsirolimus, ridaforolimus, sirolimus, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent is selected from the group consisting of amifostine, anagrelide, clodronate, neulasta, interferon, interferon alpha, folinic acid, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate, and porfimer, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent is selected from the group consisting of 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-alethine, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxy camptothecin, 16-aza-epothilone B, A105972, A204197, abiraterone, aldesleukin, acitretin, allovectin-7, altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037, AP-5280, apaziquone, apomine, aranose, arglabin, arzoxifene, atamestane, atrasentan, auristatin PE, ABT-199 (Venetoclax), ABT-263 (Navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), ARRY 162, ARRY 438162, ARRY-300, ARRY-142886 / AZD-6244 (selumetinib), ARRY-704 / AZD-8330, AR-12, AR-42, AS-703988, AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ680, AS-703026 (primasertib), avastin, AZD-2014, azacitidine, ixabepilone B, azonafide, BAY-43-9006, BAY80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, biricodar di citrate, BCX-1777, BKM-120, bliconin, BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW 2992 (afatinib, tofacitinib), BIBF 1120 (vargatef), BI 836845, BI 2536, BI 6727, BI 836845, BI 847325, BI 853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, blenoxane, bryostatins-1, bortezomib, brostallicin, busulfan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, canertinib, canfosfamide, capecitabine, carboplatin, CCl-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, ceflatonin, ceftriaxone, celecoxib, celmoleukin, cemoplatinin, CH4987655 / RO-4987655, chloranil, cilengitide, ciclosporin, CDA-II, CKD-394, CKD-602, CKI-27, clofarabine, colchicine, combretastin A4, COT inhibitor, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibody, CP-461, CV-247, cyanomorpholino doxorubicin, cytarabine, D24851, decitabine, doxorubicin, deoxyrubicin, deoxycoformycin, depsipeptide, deoxylavendustin B, dexamethasone, dexrazoxane, dienestrol, diflomotecan, didox, DMDC, dolastatin 10, dolasetron, DS-7423, E7010, E-6201, edatrexat, edotreotide, efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509, enzastaurin, enzalomed, enoxacin, epirubicin B, epratuzumab, ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine, ethynylestradiol, exatecan, exatecan mesylate, exemestane, exisulind, fenretinide, etomoxir, floxuridine, folic acid, FOLFOX, FOLFOX4, FOLFIRI, fostedil, fluoxetine, camptothecin, gallium maltolate, gefitinib, gemtuzumab, gemcitabine, GLC-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100, gp100 peptide vaccine, GSK-5126766, GSK-690693, GSK-1120212 (trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron, herceptin, hexamethylmelamine, histamine, homoharringtonine, hyaluronic acid, hydroxyurea, hydroxypregesterone caproate, ibandronate, ibritumomab tiuxetan, idatrexate, idenestrol, IDN-5109, IGF-1R inhibitors, IMC-1C11, IMC-A12 (cixutumumab), immunol, indisulam, interferon alpha-2a, interferon alpha-2b, pegylated interferon alpha-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib, ipilimumab, iproplatin, irinotecan, isohomohalichondrin-B, isoflavones, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, kahalidF, ketoconazole, KW-2170, KW-2450, lopaplatin, leflunomide, leuprorelin, leuporelin, lexidronam, LGD-1550, linezolid, lutetium texaphyrin, lomotrexol, losoxantrone, LU 223651, losoxantrone, LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechlorethamine, MEK inhibitor, MEK-162, methyltestosterone, methylprednisolone, MEDI-573, MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, midostaurin, minodronic acid, mitomycin, mivobulin, MK-2206, MK-0646 (dalozumab), MLN518, motesafam gadolinium, M S-209, MS-275, MX6, neridronate, neratinib, Nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nolatrexed, norethin, N-acetylcysteine, 06-benzylguanine, oblimersen, omeprazole, oncophage, oncoVEXGM-CSF, ormiplatin, ortataxel, OX44 antibody, OSI-027, OSI-906 (linsitinib), 4 -1BB antibody, oxantrazole, estrogen, panitumumab, patupirone, pegfilgrastim, PCK-3145, pegfilgrastim, PBI-1402, PBI-05204, PDO325901, PD-1 antibody, PEG-paclitaxel, albumin-stabilized paclitaxel, PEP-005, PF-05197281, PF-05212384, PF-04691502, PHT-427, P-04, PKC412, P54, PI-88, pelitinib Ni, pemetrexed, pentrix, perifosine, perillyl alcohol, pertuzumab, PI3K inhibitor, PI3K / mTOR inhibitor, PG-TXL, PG2, PLX-4032 / RO-5185426 (vemurafenib), PLX-3603 / RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pivaloyloxymethylbutyrate, velvet, phentodil O, PKI166, pletrexed, mithramycin, polyprenic acid, porfibrinolide, prednisone, prednisolone, quinamed, quinupristin, R115777, RAF-265, ramosetron, ranpirnase, RDEA-119 / BAY 869766, RDEA-436, rapamycin analogs, receptor tyrosine kinase (RTK) inhibitors, revimid, RG-7167, RG-7304, RG-7421, RG-7321, RG7440、rhizoxin, rhu-MAb, linifanib, risedronate, rituximab, rofecoxib, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR 109881A, rubidomycin phenylhydrazone, rubitecan, R-flurbiprofen, RX-0201, S-9788, sapphyrin, SAHA, sagostim, satraplatin, SB 408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustin, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine, suberanilohydroxamic acid, sutent, T 900607, T 138067, TAK-733, TAS-103, tacedinaline, talaporfin, tarceva, tariquitar, tasisulam, taxotere, taxoprexin, tazarotene, tegafur, temozolomide, temilifene, testosterone, testosterone propionate, temilifene, tetraplatin, tetrodotoxin, tezacitabine, thalidomide, theralux, therarubicin, thymalfasin, thymectacin, tiazofurin, tipifarnib, tirapazamine, tozasulamide, tomudex, toremofin, tracbelidin, trans-MID-107, trans-retinoic acid, traszutumab, tremelimumab, tretinoin, triacetyluridine, triapine, tricetabine, trimetrexate, TLK-286 TXD 258, tykerb / tyverb, urocidin, valrubicin, vatalanib, vincristine, vindesine, virulizin, WX-UK1, WX-554, vekatifibat, xeloda, XELOX, XL-147, XL-228, XL-281, XL-518 / R-7420 / GDC-0973, XL-765, YM-511, YM-598, ZD-4190, ZD-6474, ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZD1839, ZSTK-474, zoledronat, zosuquidar, and combinations thereof.

[0266] In one embodiment, the additional therapeutic agent comprises a steroid, including dexamethasone, prednisolone, methylprednisolone, prednisone, hydrocortisone, triamcinolone, betamethasone, and cortivazole. In one embodiment, the additional therapeutic agent comprises an antiemetic. Antiemetics include, but are not limited to, 5-HT3 receptor agonists (e.g., dolasetron, granisetron, ondansetron, tropisetron, palonosetron, and mirtazapine), dopamine agonists (e.g., domperidone, olanzapine, droperidol, haloperidol, chlorpromazine, prochlorperazine, alipride, prochlorperazine, and metoclopramide), NK1 receptor antagonists (e.g., aprepitant and casopitant), antihistamines (e.g., cyclazine, diphenhydramine, dimenhydrinate, doxylamine, meclizine, promethazine, hydroxyzine), cannabinoids (e.g., cannabis, dronabinol, nabilone, and cannabinol), benzodiazepines (e.g., midazolam and lorazepam), anticholinergics (e.g., scopolamine), trimethoprim-sulfamethoxazole, ginger, emetrol, propofol, peppermint, muscimol, and cayenne pepper.

[0267] The pharmaceutical composition can be administered to a subject via any suitable route of administration. In one embodiment, the pharmaceutical composition is administered orally, parenterally, transdermally or transmucosally. In one embodiment, the pharmaceutical composition is administered parenterally. In one embodiment, the pharmaceutical composition is administered to a subject via a parenteral route of administration intravenously (IV), subcutaneously (SC) and intramuscularly (IM). In one embodiment, the pharmaceutical composition is administered to a subject by a route of administration selected from the rectum and transdermal. In one embodiment, the pharmaceutical composition is administered to a subject in a dosage form, selecting a sterile solution, suspension, suppository, tablet and capsule. In one embodiment, the pharmaceutical composition is administered to a subject in an oral dosage form, selected from tablets, sachets, capsules, lozenges, syrups, liquids, suspensions and elixirs. In one embodiment, the pharmaceutical composition is administered to a subject in an oral dosage form, selected from tablets, hard shell capsules, soft gelatin capsules, beads, granules, aggregates, powders, gels, solids and semisolids.

[0268] In one embodiment, the pharmaceutical composition is administered to a subject as a dosage form selected from the group consisting of a sustained release form, a controlled release form, a delayed release form, and a responsive release form.

[0269] In one embodiment, the pharmaceutical composition is administered to the subject once a day. In one embodiment, the pharmaceutical composition is administered to the subject according to a less frequent (e.g., once a week or less frequently) administration regimen. In one embodiment, the pharmaceutical composition is administered to the subject according to a frequent (e.g., more than once a week) administration regimen. In one embodiment, the pharmaceutical composition is administered to the subject once a week. In one embodiment, the pharmaceutical composition is administered to the subject once every four weeks. In one embodiment, the pharmaceutical composition is administered to the subject twice a week. In one embodiment, the pharmaceutical composition is administered to the subject once every two weeks. In one embodiment, the pharmaceutical composition is administered to the subject once every three weeks. In one embodiment, the pharmaceutical composition is administered to the subject in a repeating cycle of once a week, once every two weeks, once every three weeks, once every four weeks, or a combination thereof.

[0270] In one embodiment, the method of treatment comprises administering to a subject in need of such treatment: (i) a first therapeutic agent comprising a compound comprising e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in combination with (ii) a second therapeutic agent, wherein the first therapeutic agent and the second therapeutic agent are administered simultaneously or sequentially; and further comprising assaying a biological sample for expression of an endoplasmic reticulum (ER) stress response gene. In one embodiment, the endoplasmic reticulum stress response gene is selected from the group comprising, but not limited to, C / EBP-homologous protein (CHOP), activating transcription factor 3 (ATF3), and both CHOP and ATF3. In one embodiment, the endoplasmic reticulum stress response gene is selected from the group comprising, but not limited to, ATF3, activating transcription factor 4 (ATF4), CHOP, IRE1, immunoglobulin binding protein (BiP), eukaryotic translation initiation factor 2A (eIF2a), X-box binding protein 1 (XBP1). The biological sample can be a tumor, peripheral blood mononuclear cells, or a skin biopsy. The biological sample can be obtained prior to, during, or after administration of the drug. In one embodiment, the method of treatment further comprises adjusting the dosage of the first therapeutic agent to achieve about 50%, 75%, 100%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 525%, 550%, 575%, 600%, or greater than 600% induction of one or more ER stress genes. In one embodiment, the method of treatment further comprises adjusting the dosage of the first therapeutic agent to achieve about 50% to about 100%, about 100% to about 150%, about 150% to about 200%, about 200% to about 250%, about 250% to about 300%, about 300% to about 350%, about 350% to about 400%, about 400% to about 450%, about 450% to about 500%, about 500% to about 550%, about 550% to about 600%, or greater than 600% induction of ER stress genes. In one embodiment, the method of treatment further comprises adjusting the dosage of the first therapeutic agent to achieve about 50% to about 100%, about 100% to about 200%, about 200% to about 300%, about 300% to about 400%, about 400% to about 500%, about 500% to about 600%, or greater than 600% induction of ER stress genes.

[0271] In one embodiment, the method of treatment comprises administering to a subject in need of such treatment: (i) a first therapeutic agent comprising a compound comprising imirichne, e.g., ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in combination with (ii) a second therapeutic agent, wherein the first therapeutic agent and the second therapeutic agent are administered simultaneously or sequentially; and further comprising assaying the expression of proteasome activity in a biological sample. In one embodiment, the proteasome activity can be chymotrypsin-like, trypsin-like, and / or caspase-like activity. In one embodiment, the biological sample can be a tumor, peripheral blood mononuclear cells, or a skin biopsy. The biological sample can be obtained prior to, during, or after administration of the drug. In one embodiment, the method of treatment further comprises adjusting the dosage to achieve an inhibition of about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% of proteasome activity. In one embodiment, the method of treatment further comprises adjusting the dosage to achieve an inhibition of at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% of proteasome activity. In one embodiment, the method of treatment further comprises adjusting the dosage to achieve an inhibition of at least about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or greater than 90% of proteasome activity.

[0272] In one aspect, provided herein is a method of treatment comprising administering to a subject in need of such treatment a combination of a first therapeutic agent comprising imirichne, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof (e.g., a disalt or trisalt), and a second therapeutic agent, the method comprising:

[0273] (i) administering to the subject the first therapeutic agent;

[0274] (ii) waiting for a predetermined waiting time after the time the first therapeutic agent is administered to the subject; and / or until an adverse event has resolved or is resolving; and

[0275] (iii) administering a second therapeutic agent to the subject, wherein the predetermined waiting period is selected so as to obtain a delayed therapeutic effect of the first therapeutic agent without increasing the risk of possible combined toxic effect of the first and second therapeutic agents. In one embodiment, the predetermined waiting period is determined based on the clearance rate of the first therapeutic agent compound or its metabolite. In one embodiment, the predetermined waiting period is determined by quantitatively assessing renal function and renal parameters. In one embodiment, the predetermined waiting period is determined by a determination of renal function, wherein the determination is selected from the group consisting of: serum level of the first therapeutic agent compound or its metabolite; clearance rate of the first therapeutic agent compound or its metabolite; 24 hour urine clearance of the first therapeutic agent compound or its metabolite.

[0276] In one embodiment of the method of treatment, the predetermined waiting period is substantially equal to the time required for systemic clearance of the first therapeutic agent compound or its metabolite from the subject. In one embodiment of the method of treatment, the predetermined waiting period is substantially equal to the time required for renal clearance of the first therapeutic agent compound or its metabolite from the subject. In one embodiment of the method of treatment, the predetermined waiting period is substantially equal to the time required for hepatic clearance of the first therapeutic agent compound or its metabolite from the subject. In one embodiment of the method of treatment, the predetermined waiting period is substantially equal to the time required for total clearance of the first therapeutic agent compound or its metabolite from the subject. In one embodiment of the method of treatment, the predetermined waiting period is about 4 hours. In other embodiments, the waiting period is 1 day. In one embodiment, the waiting period is until the C max has elapsed. In other embodiments, the waiting period is after most adverse events have resolved or are resolving. In one embodiment of the method of treatment, the predetermined waiting period is about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, or about 7 days. In one embodiment of the method of treatment, the predetermined waiting period is in the range of about 1-7 days, about 1-6 days, about 1-5 days, about 1-4 days, about 1-3 days, or about 1-2 days. In one embodiment, the waiting period is up to 3 weeks. The foregoing is considered a "treatment time period".

[0277] When the order of administration is reversed, the time of administration of the first therapeutic agent can be after the C max of the first administered drug has elapsed. In one embodiment, the administration of the first therapeutic agent can be after most or substantially all of the first administered drug has been eliminated from the body or the toxic effects of the first administered drug have resolved or are resolving.

[0278] In one embodiment, the method of treatment further comprises monitoring the level of the first therapeutic agent compound or metabolite thereof in the subject using pharmacokinetic profiling. In some such embodiments, monitoring the level of the first therapeutic agent compound or metabolite thereof in the subject using pharmacokinetic profiling comprises constructing a pharmacokinetic profile of the first therapeutic agent compound or metabolite thereof using concentrations of the first therapeutic agent compound or metabolite thereof in at least two samples obtained from the subject at time points suitable for constructing a pharmacokinetic profile. In one embodiment of monitoring the level of the first therapeutic agent compound or metabolite thereof in the subject using pharmacokinetic profiling, samples are collected from the subject at the point of care or point of use by sampling or self-sampling on a point of care or point of use device or on a substrate suitable for storing the sample, and then quantified in a laboratory. In one embodiment, each point of care or point of use device is capable of quantifying the first therapeutic agent compound or metabolite thereof. In one embodiment of monitoring the level of the first therapeutic agent compound or metabolite thereof in the subject, one or more samples are collected from the subject at the point of care or point of use by biopsy device for analysis at the point of care or point of use device or for storage prior to analysis by a laboratory. In one embodiment, the biopsy is taken after a 3-8 hour time interval after administration of the first therapeutic agent to the subject. In one embodiment, the biopsy is taken after a 3-24 hour time interval after administration of the first therapeutic agent to the subject. In one embodiment, the biopsy is taken after an 8-24 hour time interval after administration of the first therapeutic agent to the subject. In one embodiment, the biopsy is taken after a 2 day time interval after administration of the first therapeutic agent to the subject. In one embodiment, the biopsy is taken after a 3 day time interval after administration of the first therapeutic agent to the subject. In one embodiment, the biopsy is taken after a 4 day time interval after administration of the first therapeutic agent to the subject. In one embodiment, the biopsy is taken after a 1-7 day time interval after administration of the first therapeutic agent.

[0279] In one embodiment, the pharmacokinetic profile comprises pharmacokinetic parameters suitable for guiding the pharmacokinetic profile of the first therapeutic agent in the subject being treated. In one embodiment of the method of treatment, C max In one embodiment, the Cmax is in the range from about 1000 ng / dL to 1500 ng / dL for a certain treatment period. In one embodiment, the Cmax is in the range from about 1000 ng / dL to 1500 ng / dL for a certain treatment period. max In one embodiment, the Cmax is less than 1500 ng / dL and greater than 85 ng / dL for a certain treatment period. In one embodiment, the Cmax is less than 1500 ng / dL and greater than 85 ng / dL for a certain treatment period. max In one embodiment, the Cmax is in the range from about 1000 ng / mL to 1500 ng / mL for a certain treatment period. In one embodiment, the Cmax is in the range from about 1000 ng / mL to 1500 ng / mL for a certain treatment period. max In one embodiment, the Cmax is less than 1500 ng / mL and greater than 85 ng / mL for a certain treatment period.

[0280] In one embodiment, the maximum concentration ("C max ") of the first therapeutic agent in the blood (whole blood, plasma, or serum) of the subject after administration of the first therapeutic agent to the subject is C max from about 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, or 1490 ng / dL to about 1500 ng / dL; from about 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 ng / dL to about 150 ng / dL; or from about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14, or 14.5 ng / dL to about 15 ng / dL.

[0281] In one embodiment, the maximum concentration ("C max ") of the first therapeutic agent in the blood (whole blood, plasma, or serum) of the subject after administration of the first therapeutic agent to the subject is C maxFrom about 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 126 0, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, or 1490 ng / mL to about 1500 ng / mL; from about 1 00, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 1 14.5 ng / mL to about 15 ng / mL; or from about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14, or 14.5 ng / mL to about 15 ng / mL.

[0282] In one embodiment, the maximum concentration (“C max ”) is selected from about 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280 , 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480 or 1490 ng / dL. In one embodiment, the C in the blood (whole blood, plasma, or serum) of the subject after administration of the first therapeutic agent is max (“C maxabout 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 ng / dL. In one embodiment, the C max about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14, or 14.5 ng / dL.

[0283] In one embodiment, the C max about 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, or 1490 ng / mL. In one embodiment, the C max about 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 ng / mL. In one embodiment, the C max about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14, or 14.5 ng / mL.

[0284] In one embodiment, the C maxselected from about 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 765, 775, 785, 795, 805, 815, 825, 835, 845, 855, 865, 875, 885, 895, 905, 915, 925, 935, 945, 955, 965, 975, 985, 995, 1005, 1015, 1025, 1035, 1045, 1055, 1065, 1075, 1085, 1095, 1105, 1115, 1125, 1135, 1145, 1155, 1165, 1175, 1185, 1195, 1205, 1215, 1225, 1235, 1245, 1255, 1265, 1275, 1285, 1295, 1305, 1315, 1325, 1335, 1345, 1355, 1365, 1375, 1385, 1395, 1405, 1415, 1425, 1435, 1445, 1455, 1465, 1475, 1485, 1495, or 1500 ng / dL. In one embodiment, the first therapeutic agent is administered at a dose of about 1000 ng / dL. maxabout 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, or 14.5 ng / dL. In one embodiment, the C max about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, or 14.5 ng / dL. In one embodiment, the C

[0285] about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, or 14.5 ng / dL. In one embodiment, the C maxselected from about 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 765, 775, 785, 795, 805, 815, 825, 835, 845, 855, 8 65, 875, 885, 895, 905, 915, 925, 935, 945, 955, 965, 975, 985, 995, 1005, 1015, 1025, 1035, 1045, 1055, 1065, 1075, 1085, 1095, 1105, 1115, 1125, 1135, 1145, 1155, 1165, 1175, 1185, 1195, 12 05, 1215, 1225, 1235, 1245, 1255, 1265, 1275, 1285, 1295, 1305, 1315, 1325, 1335, 1345, 1355, 1365, 1375, 1385, 1395, 1405, 1415, 1425, 1435, 1445, 1455, 1465, 1475, 1485, 1495 or 1500 ng / mL. In one embodiment, the C maxabout 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 ng / mL. In one embodiment, the C max about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, or 14.5 ng / mL.

[0286] In one embodiment, the C max ranging from about 85 ng / dL to 1500 ng / dL; from about 8.5 ng / dL to 150 ng / dL; or from about 0.85 ng / dL to 15 ng / dL. In one embodiment, the C maxselected from about 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 765, 775, 785, 795, 805, 815, 825, 835, 845, 855, 865, 875, 885, 895, 905, 915, 925, 935, 945, 955, 965, 975, 985, 995, 1005, 1015, 1025, 1035, 1045, 1055, 1065, 1075, 1085, 1095, 1105, 1115, 1125, 1135, 1145, 1155, 1165, 1175, 1185, 1195, 1205, 1215, 1225, 1235, 1245, 1255, 1265, 1275, 1285, 1295, 1305, 1315, 1325, 1335, 1345, 1355, 1365, 1375, 1385, 1395, 1405, 1415, 1425, 1435, 1445, 1455, 1465, 1475, 1485, or 1495 ng / dL to about 1500 ng / dL;about 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 ng / dL to about 150 ng / dL; or about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, or 14.5 ng / dL to about 15 ng / dL.

[0287] In one embodiment, the Cmaxof the first therapeutic agent after administration of the first therapeutic agent is in the range from about 85 ng / mL to 1500 ng / mL; from about 8.5 ng / mL to 150 ng / mL; or from about 0.85 ng / mL to 15 ng / mL. max In one embodiment, the Cmaxof the first therapeutic agent after administration of the first therapeutic agent is in the range from about 85 ng / mL to 1500 ng / mL; from about 8.5 ng / mL to 150 ng / mL; or from about 0.85 ng / mL to 15 ng / mL. maxselected from about 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 765, 775, 785, 795, 805, 815, 825, 835, 845, 855, 865, 875, 885, 895, 905, 915, 925, 935, 945, 955, 965, 975, 985, 995, 1005, 1015, 1025, 1035, 1045, 1055, 1065, 1075, 1085, 1095, 1105, 1115, 1125, 1135, 1145, 1155, 1165, 1175, 1185, 1195, 1205, 1215, 1225, 1235, 1245, 1255, 1265, 1275, 1285, 1295, 1305, 1315, 1325, 1335, 1345, 1355, 1365, 1375, 1385, 1395, 1405, 1415, 1425, 1435, 1445, 1455, 1465, 1475, 1485, or 1495 ng / mL to about 1500 ng / mL;about 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 ng / mL to about 150 ng / mL; or about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, or 14.5 ng / mL to about 15 ng / mL.

[0288] In one embodiment, the total drug exposure over time is measured as the area under the curve ("AUC") of the concentration of the drug in the blood (whole blood, plasma, or serum) plotted against time after administration of the drug, ranging from about 150 ng hr / mL to about 8000 ng hr / mL; from about 15 ng hr / mL to about 800 ng hr / mL; or from about 1.5 ng hr / mL to about 80 ng hr / mL. In one embodiment, the AUC is less than 8000 ng hr / mL and greater than or equal to 150 ng hr / mL. In one embodiment, the AUC is less than 800 ng hr / mL and greater than or equal to 15 ng hr / mL. In one embodiment, the AUC is less than 80 ng hr / mL and greater than or equal to 1.5 ng hr / mL.

[0289] In one embodiment, the total drug exposure over time is AUC from about 100 ng hr / mL to about 8000 ng hr / mL; from about 10 ng hr / mL to about 800 ng hr / mL; or from about 1 ng hr / mL to about 80 ng hr / mL. In one embodiment, the total drug exposure over time is AUC from about 150, 200, 400, 600, 800, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, 4000, 4200, 4400, 4600, 4800, 5000, 5200, 5400, 5600, 5800, 6000, 6200, 6400, 6600, 6800, 7000, 7200, 7400, 7600, or 7800 ng hr / mL to about 8000 ng hr / mL. In one embodiment, the total drug exposure over time is AUC from about 15, 20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560, 580, 600, 620, 640, 660, 680, 700, 720, 740, 760, or 780 ng hr / mL to about 800 ng hr / mL. In one embodiment, the total drug exposure over time is AUC from about 1.5, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, or 78 ng hr / mL to about 80 ng hr / mL.

[0290] In one embodiment, the total drug exposure over time is AUC from about 100 ng hr / mL to about 8000 ng hr / mL, from about 10 ng hr / mL to about 800 ng hr / mL; or from about 1 ng hr / mL to about 80 ng hr / mL. In one embodiment, the total drug exposure over time is AUC from about 150 ng hr / mL to about 7800, 7600, 7400, 7200, 7000, 6800, 6600, 6400, 6200, 6000, 5800, 5600, 5400, 5200, 5000, 4800, 4600, 4400, 4200, 4000, 3800, 3600, 3400, 3200, 3000, 2800, 2600, 2400, 2200, 2000, 1800, 1600, 1400, 1200, 1000, 800, 600, 400, or 200 ng hr / mL. In one embodiment, the total drug exposure over time is AUC from about 15 ng hr / mL to about 780, 760, 740, 720, 700, 680, 660, 640, 620, 600, 580, 560, 540, 520, 500, 480, 460, 440, 420, 400, 380, 360, 340, 320, 300, 280, 260, 240, 220, 200, 180, 160, 140, 120, 100, 80, 60, 40, or 20 ng hr / mL. In one embodiment, the total drug exposure over time is AUC from about 1.5 ng hr / mL to about 78, 76, 74, 72, 70, 68, 66, 64, 62, 60, 58, 56, 54, 52, 50, 48, 46, 44, 42, 40, 38, 36, 34, 32, 30, 28, 26, 24, 22, 20, 18, 16, 14, 12, 10, 8, 6, 4, or 2 ng hr / mL. In one embodiment, the total drug exposure over time is AUC from about 100 ng hr / mL to about 200 ng hr / mL; from about 10 ng hr / mL to about 20 ng hr / mL; or from about 1 ng hr / mL to about 2 ng hr / mL.

[0291] In one embodiment, the total drug exposure over time is AUC selected from the group consisting of about 100, 150, 200, 400, 600, 800, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, 4000, 4200, 4400, 46000, 4800, 5000, 5200, 5400, 5600, 5800, 6000, 6200, 6400, 6600, 6800, 7000, 7200, 7400, 7600, 7800, and 8000 ng hr / mL. In one embodiment, the total drug exposure over time is AUC selected from the group consisting of about 10, 15, 20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 4600, 480, 500, 520, 540, 560, 580, 600, 620, 640, 660, 680, 700, 720, 740, 760, 780, and 800 ng hr / mL. In one embodiment, the total drug exposure over time is AUC selected from the group consisting of about 1, 15, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 460, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, and 80 ng hr / mL.

[0292] In another aspect, provided herein are methods of treating or using a composition to treat a disease state, the methods comprising administering to a subject in need of such treatment a combination of a first therapeutic agent and a second therapeutic agent, the methods comprising:

[0293] (i) administering to the subject a first therapeutic agent comprising imirichne, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof;

[0294] (ii) monitoring the level of the first therapeutic agent compound or a metabolite thereof in the subject using pharmacokinetic profiling; and

[0295] (iii) administering the second therapeutic agent conditioned on the level of the first therapeutic agent in the subject. In one embodiment, the monitoring step comprises constructing a pharmacokinetic profile of the first therapeutic agent compound or metabolite thereof for the subject using the concentrations of the first therapeutic agent compound or metabolite thereof in a plurality of samples obtained from the subject at time points suitable for constructing a pharmacokinetic profile. In one embodiment, the samples are collected at the point-of-care or point-of-use by sampling or self-sampling on a point-of-care or point-of-use device or on a substrate suitable for storing the sample, and then the compound or metabolite thereof is quantified by a laboratory. In one embodiment, each point-of-care or point-of-use device is capable of quantifying the compound or metabolite thereof. In one embodiment, the pharmacokinetic profile comprises pharmacokinetic parameters suitable for guiding administration of the compound or salt thereof to the subject. In one embodiment, the samples comprise 2-12 samples. In one embodiment, the samples are collected over a period of up to 8 hours, up to 24 hours, up to 48 hours, or up to 72 hours. In one embodiment, the pharmacokinetic parameters comprise at least one parameter selected from the group consisting of AUC, AUC inf , T max , C max , time over threshold, steady state concentration, rate of absorption, rate of clearance, rate of distribution, terminal T-l / 2, or a parameter derived from non-compartmental pharmacokinetic (PK) or compartmental PK analysis, including physiologically-based compartmental PK analysis. In one embodiment, the method of treatment further comprises generating a report comprising the pharmacokinetic profile of the subject. In one embodiment, the report comprises recommendations based on the pharmacokinetic profile of the subject regarding administration. In one embodiment, based on one or more pharmacokinetic parameters, it is indicated to reduce the dose of ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof to reduce the risk of toxicity. In one embodiment, the reduction of the dose of the compound or salt thereof is indicated based on a time over threshold indicating, wherein the threshold is a drug concentration over which toxicity occurs, or one or more of: AUC, AUC inf , mean residence time (MRT), an index defining the pharmacokinetic profile, steady state volume of distribution (Vss), terminal volume of distribution (Vz), or a combination of a set of pharmacokinetic variables sufficient to describe the pharmacokinetic profile. In one embodiment, based on one or more pharmacokinetic parameters, it is indicated to adjust the dose of the compound or salt thereof to increase efficacy. In one embodiment, the adjustment of the dose of the compound or salt thereof is based on one or more of: AUC, AUC infMRT, an index defining the pharmacokinetic profile, steady state volume of distribution (Vss), volume of distribution at terminal phase (Vz), or a combination of a set of pharmacokinetic variables that sufficiently describe the pharmacokinetic profile, indicates an increase in the dose of the compound or salt thereof. In one embodiment, the dose of the compound or salt thereof is adjusted to within 5% to 25% of a predetermined target value. In one embodiment, each sample is applied to a point-of-care device or point-of-use device to determine the concentration of the compound or metabolite thereof, wherein the point-of-care device or point-of-use device comprises a lateral flow test strip having a structure and composition such that application of one or more samples to the lateral flow test strip results in a portion of the drug in the sample binding to a component of the lateral flow test strip, thereby producing a detectable signal that is directly proportional to the concentration of the drug in the applied sample. In one embodiment, the sample is applied to a matrix suitable for storing the sample prior to quantification in a laboratory. In one embodiment, the sample is stored as a dried blood spot. In one embodiment, the drug concentration is measured by ELISA, LC MS MS, LC UV, or LCMS. In one embodiment, the pharmacokinetic parameters include at least one of steady state concentration, absorption, and terminal T 1 / 2 In one embodiment, at least one of the samples is whole blood.

[0296] IX. Multimodal treatment methods

[0297] In one aspect, provided herein is a multimodal treatment method in which a subject in need of such treatment is administered amisulpride, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof supplemented by administration of other treatment modalities. In one embodiment, the multimodal treatment method comprises administering to a subject in conjunction with radiation therapy or after a determination that radiation is no longer effective, a pharmaceutical composition comprising amisulpride, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the multimodal treatment method comprises administering to a subject in conjunction with radiation therapy, a pharmaceutical composition comprising amisulpride, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition comprising amisulpride, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof and the radiation therapy are administered simultaneously or sequentially in any order. In one embodiment, the multimodal treatment method comprises administering to a subject in sequential arrangement, a pharmaceutical composition comprising amisulpride, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, in conjunction with radiation therapy. In one embodiment, the multimodal treatment method comprises administering to a subject in need of such treatment, a pharmaceutical composition comprising amisulpride, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, simultaneously with radiation therapy. In one embodiment, the multimodal treatment method is used to treat cancer. In one embodiment, the multimodal treatment method comprises administering to a subject in need of such treatment, a cancer, a pharmaceutical composition comprising amisulpride, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof and irradiating the cancer cells with a radiation beam. In one embodiment, the multimodal treatment method uses conformal radiation therapy (CRT) techniques to deliver the dose volume histogram (DVH) prescribed for the cancer subject. In one embodiment, the multimodal treatment method uses intensity modulated radiation therapy (IMRT) techniques to deliver radiation to the cancer cells. In one embodiment, the multimodal treatment method uses techniques that compensate for tumor motion in the subject during treatment (e.g., in the case of a chest tumor that must be administered a radiation dose that moves with the patient’s breath). For example, the multimodal treatment method uses scanning techniques of four-dimensional computed tomography (4D CT) to adjust the radiation field delivered to compensate for tumor motion in the respiratory cycle.

[0298] Any suitable radiation type, including fractionated gamma radiation, IMRT (intensity modulated radiation therapy), gamma knife, proton therapy, and brachytherapy can be used with the multi-modal treatment method. Radiation therapy and administration of an imirichne, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof can be used to treat brain tumors, e.g., glioblastoma or disease that has metastasized from lung cancer to the brain. The multi-modal treatment method can be used to treat lung cancer, pancreatic cancer, rectal cancer, breast cancer, sarcoma, prostate cancer, gynecological malignancies, and lymphoma. Gamma knife is often used to treat brain metastases. In one embodiment, the multi-modal treatment method includes the use of proton therapy to treat cancer, including brain tumors, prostate cancer, and any tumor near vital organs, in the latter case it is very important to minimize toxicity to nearby normal tissue.

[0299] In one embodiment, the multi-modal treatment method includes simultaneous or combined administration of a pharmaceutical composition comprising an imirichne, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, with adoptive cell therapy (e.g., CAR-T (JCAR 14, 15, 16, 17, KTE-C19, or CTL019), other T cells (AFM13), or NK (CDNO-109 or NK-92)) to a cancer subject in need of such treatment.

[0300] In one embodiment, the multi-modal treatment method eliminates minimal residual disease without increasing toxicity resulting from treatment with an imirichne, e.g., ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the multi-modal treatment method improves prognosis and / or reduces adverse side effects associated with the disease state or condition in the subject receiving treatment.X. Other imirichne derivatives, analogs, and salts

[0301] In one aspect, provided herein are analogs of the compound of formula (10) and methods of making the same. It will be understood by those skilled in the art that the general principles and concepts described above in connection with ONC201 and the compound of formula (10) and salts thereof, including those related to methods and pharmaceutical compositions, apply with equal force to the analogs below and salts thereof.

[0302] In one embodiment, the analog has the structure of compound (25):

[0303] wherein Y is NR4or O, and wherein R1, R2, R3, and R4independently represent hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and heterocyclic radicals. In one embodiment, R1, R1, R2, R3, and R4are optionally substituted. In one embodiment, some or all of the hydrogen atoms in R1, R1, R2, R3, and R4are replaced with deuterium. In other embodiments, R1, R2, R3, and R4are independently selected from the group consisting of H, C 1-4 alkyl, C 1-4 alkylphenyl, C 1-4 alkylphenyl ketone, C 1-4 benzylpiperazine, and C 1-4 alkylthiophenyl, wherein C 1-4 alkyl, C 1-4 alkylphenyl, C 1-4 alkylphenyl ketone, and C 1-4 benzylpiperazine are optionally substituted with C 1-4 alkyl, hydroxyl, or halogen. In yet other embodiments, R1, R2, R3, and R4are independently selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzylpiperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph.

[0304] In one embodiment, the analog has the structure of compound (26):

[0305] wherein R1and R2independently represent H, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and heterocyclic radicals. In one embodiment, R1and R2are independently selected from the group consisting of H, C 1-4 alkyl, C 1-4 alkylphenyl, C 1-4 alkylphenyl ketone, C 1-4 benzylpiperazine, and C 1-4 alkylthiophenyl, wherein C 1-4 alkyl, C 1-4 alkylphenyl, C1-4 alkyl phenyl ketone and C 1-4 benzyl piperazine is optionally substituted with C 1-4 alkyl, C 1-4 alkoxy, hydroxy, perhalo C 1-4 alkyl or halo. In one embodiment, R1is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzyl piperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph. In one embodiment, R2is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzyl piperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph.

[0306] In one embodiment, R1is benzyl optionally substituted with one or more of the following substituents, individually or in combination, at the ortho, meta, and / or para position of the benzyl ring: -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R2is benzyl optionally substituted with one or more of the following substituents, individually or in combination, at the ortho, meta, and / or para position of the benzyl ring: -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen.

[0307] In one embodiment, R1is H. In one embodiment, R1is substituted or unsubstituted arylalkyl, such as benzyl or phenethyl. In one embodiment, the arylalkyl is substituted with C 1-4 alkyl, C 1-4 alkoxy, hydroxy, perhalo C 1-4 alkyl or halo.

[0308] In one embodiment, R2is substituted or unsubstituted arylalkyl, such as benzyl or phenethyl. In one embodiment, the arylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3, and -OCH3. In one embodiment, R2is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In one embodiment, R2is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3, and -OCH3. 1-4 alkyl, C 1-4 alkyl, C 1-4 alkyl, or halo. In one embodiment, the arylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3, and -OCH3. In one embodiment, R2is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In one embodiment, R2is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3, and -OCH3. 1-4 alkyl, C 1-4 alkyl, C 1-4 alkyl, or halo. In one embodiment, the arylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3, and -OCH3. In one embodiment, R2is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In one embodiment, R2is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3, and -OCH3.

[0309] In one embodiment, the analog has the structure of compound (27):

[0310]

[0311] wherein R1is H, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and heterocyclic radicals. In one embodiment, R1is selected from the group consisting of H, C 1-4 alkyl, C 1-4 alkyl, C 1-4 alkyl, C 1-4 alkyl, and C 1-4 alkyl, and C 1-4 alkyl, C 1-4 alkyl, C 1-4 alkyl, and C 1-4 alkyl, and C 1-4 alkyl, C 1-4 alkyl, C 1-4alkyl or halo-substituted. In one embodiment, R1is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzylpiperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph.

[0312] In one embodiment, R1is benzyl optionally substituted at the ortho, meta, and / or para position of the benzyl ring with one or more of the following substituents, alone or in combination: -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 where p is an integer from 2 to 20 and where X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R1is H. In one embodiment, R1is substituted or unsubstituted arylalkyl, such as benzyl or phenethyl. In one embodiment, the arylalkyl is substituted with C 1-4 alkyl, C 1-4 alkoxy, hydroxy, perhaloC 1-4 alkyl or halo-substituted.

[0313] In one embodiment, the analog has the structure of compound (28):

[0314]

[0315] where R1and R2are H, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and heterocyclic radical. In one embodiment, R1and R2are independently selected from the group consisting of H, C 1-4 alkyl, C 1-4 alkylphenyl, C 1-4 alkylphenyl ketone, C 1-4 benzylpiperazine, and C 1-4 alkylthienyl, where C 1-4 alkyl, C 1-4 alkylphenyl, C 1-4 alkylphenyl ketone, and C 1-4 benzylpiperazine is optionally substituted with C 1-4alkyl, C 1-4 alkoxy, hydroxy, perhalo C 1-4 alkyl or halo. In one embodiment, R1is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, CH2CH2(4-N-benzylpiperazine), and (CH2)3CO-4F-Ph. In one embodiment, R2is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzylpiperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph. In one embodiment, when R1is CH2Ph, R2is not CH2-(2-CH3-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(2-CH3-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(2,4-diF-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(4-CF3-Ph).

[0316] In one embodiment, R1is benzyl optionally substituted at the ortho, meta, and / or para position of the benzyl ring by one or more, alone or in combination, of -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R2is benzyl optionally substituted at the ortho, meta, and / or para position of the benzyl ring by one or more, alone or in combination, of -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen.

[0317] In one embodiment, R1 is H. In one embodiment, R1 is substituted or unsubstituted arylalkyl, such as benzyl or phenethyl. In one embodiment, arylalkyl is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0318] In one embodiment, R2 is a substituted or unsubstituted arylalkyl group, such as a benzyl or phenethyl group. In one embodiment, the arylalkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, the arylalkyl group is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3. In one embodiment, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In one embodiment, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl group is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halo substituted. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0319] In one embodiment, the analog has the structure of compound (29):

[0320]

[0321] wherein R1 and R2 independently represent H, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, aralkylthio, alkanoyl, sulfhydryl, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and heterocyclic radicals. In one embodiment, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzylpiperazine and C 1-4 The group consisting of alkylthiophene groups, wherein C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone and C 1-4 Benzylpiperazine is optionally C1-4 alkyl, C 1-4 alkoxy, hydroxy, perhalo C 1-4 alkyl or halo. In one embodiment, R1is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzylpiperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph. In one embodiment, R2is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzylpiperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph. In one embodiment, when R1is CH2Ph, R2is not CH2-(2-CH3-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(2-CH3-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(2,4-diF-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(4-CF3-Ph).

[0322] In one embodiment, R1is benzyl optionally substituted at the ortho, meta, and / or para position of the benzyl ring by one or more, alone or in combination, of -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R2is benzyl optionally substituted at the ortho, meta, and / or para position of the benzyl ring by one or more, alone or in combination, of -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen.

[0323] In one embodiment, R1 is H. In one embodiment, R1 is substituted or unsubstituted arylalkyl, such as benzyl or phenethyl. In one embodiment, arylalkyl is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0324] In one embodiment, R2 is a substituted or unsubstituted arylalkyl group, such as a benzyl or phenethyl group. In one embodiment, the arylalkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, the arylalkyl group is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3. In one embodiment, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In one embodiment, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl group is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halo substituted. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0325] In one embodiment, the analog has the structure of compound (30):

[0326]

[0327] wherein R1 and R2 independently represent H, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, aralkylthio, alkanoyl, sulfhydryl, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and heterocyclic radicals. In one embodiment, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzylpiperazine and C 1-4 The group consisting of alkylthiophene groups, wherein C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone and C 1-4 Benzylpiperazine is optionally C1-4 alkyl, C 1-4 alkoxy, hydroxy, perhalo C 1-4 alkyl or halo. In one embodiment, R1is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzylpiperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph. In one embodiment, R2is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzylpiperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph. In one embodiment, when R1is CH2Ph, R2is not CH2-(2-CH3-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(2-CH3-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(2,4-diF-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(4-CF3-Ph).

[0328] In one embodiment, R1is benzyl optionally substituted at the ortho, meta, and / or para position of the benzyl ring by one or more, alone or in combination, of -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R2is benzyl optionally substituted at the ortho, meta, and / or para position of the benzyl ring by one or more, alone or in combination, of -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen.

[0329] In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl group, such as a benzyl or phenethyl group. In one embodiment, the arylalkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0330] In one embodiment, R2 is a substituted or unsubstituted arylalkyl group, such as a benzyl or phenethyl group. In one embodiment, the arylalkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, the arylalkyl group is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3. In one embodiment, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In one embodiment, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl group is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halo substituted. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0331] In one embodiment, the analog has the structure of compound (31):

[0332]

[0333] wherein R1 and R2 independently represent H, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, aralkylthio, alkanoyl, sulfhydryl, alkylthio, arylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and heterocyclic radicals. In one embodiment, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzylpiperazine and C 1-4 The group consisting of alkylthiophene groups, wherein C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone and C 1-4benzylpiperazine is optionally substituted with C 1-4 alkyl, C 1-4 alkoxy, hydroxy, perhalo C 1-4 alkyl or halo. In one embodiment, R1is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzylpiperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph. In one embodiment, R2is selected from the group consisting of H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzylpiperazine), CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, and (CH2)3CO-4F-Ph. In one embodiment, when R1is CH2Ph, R2is not CH2-(2-CH3-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(2-CH3-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(2,4-diF-Ph). In one embodiment, R1is CH2Ph and R2is CH2-(4-CF3-Ph).

[0334] In one embodiment, R1is benzyl optionally substituted with one or more of the following substituents, individually or in combination, at the ortho, meta, and / or para position of the benzyl ring: -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R2is benzyl optionally substituted with one or more of the following substituents, individually or in combination, at the ortho, meta, and / or para position of the benzyl ring: -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, or -OC p X 2p+1 wherein p is an integer from 2 to 20 and wherein X is a halogen.

[0335] In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl group, such as a benzyl or phenethyl group. In one embodiment, the arylalkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0336] In one embodiment, R2 is a substituted or unsubstituted arylalkyl group, such as a benzyl or phenethyl group. In one embodiment, the arylalkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, the arylalkyl group is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3. In one embodiment, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In one embodiment, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl group is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halo substituted. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0337] In one embodiment, provided herein is a compound of formula (100): Wherein R1 and R2 are independently selected from H, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, aralkylthio and acyl radical. In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2-CH3-Ph), which is ONC201 linear isomer (i.e., TIC-10) TIC-10 lacks anticancer activity (Jacob et al., Angew. Chem. Int. Ed., (2014) 53: 6628; Wagner et al., Oncotarget (2015) 5(24): 12728). However, as shown in the Examples, TIC-10 is a CXCR7 agonist. CXCR7 agonists can be used for liver regeneration and the prevention or treatment of liver fibrosis (Nature (2014) 505: 97).

[0338] In one embodiment, R1 and R2 are independently selected from H, C 1-4 Alkyl, C1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzylpiperazine, C 1-4 Alkylthienyl, C 1-4 Alkylpyridyl, C 1-4 Alkylisoxazolidinyl, C 1-4 Alkylmorpholinyl, C 1-4 Alkylthiazolyl and C 1-4 The group consisting of alkylpyrazinyl, wherein C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzylpiperazine, C 1-4 Alkylthienyl, C 1-4 Alkylpyridyl, C 1-4 Alkylisoxazolidinyl, C 1-4 Alkylmorpholinyl, C 1-4 Alkylthiazolyl and C 1-4 The alkylpyrazinyl group is optionally C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, R1 and / or R2 are substituted or unsubstituted arylalkyl or heteroarylalkyl. In one embodiment, heteroarylalkyl is selected from C 1-4 Alkylpyrrolyl, C 1-4 Alkyl furanyl, C 1-4 Alkylpyridyl, C 1-4 Alkyl-1,2,4-thiadiazolyl, C 1-4 Alkyl pyrimidinyl, C 1-4 Alkylthienyl, C 1-4 Alkylisothiazolyl, C 1-4 Alkyl imidazolyl, C 1-4 Alkyl tetrazolyl, C 1-4 Alkylpyrazinyl, C 1-4 Alkyl pyrimidinyl, C 1-4 Alkylquinolinyl, C 1-4 Alkylisoquinolinyl, C 1-4 Alkylthiophenyl, C 1-4 Alkylbenzothiophene, C 1-4 Alkylisobenzofuranyl, C 1-4 Alkylpyrazolyl, C 1-4 Alkyl indolyl, C 1-4 Alkylpurine, C 1-4 Alkylcarbazolyl, C 1-4 Alkylbenzimidazole and C 1-4 Alkylisoxazolyl.

[0339] In one embodiment, R1and / or R2is benzyl optionally substituted on the benzyl ring with one or more of the following substituents: X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, -OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m , and C(O)OR m ; R m and R n are independently selected from H or C1-C4alkyl; and wherein p is an integer from 2 to 20 and wherein X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl.

[0340] XI. Examples

[0341] It is to be understood that the following description and examples are only meant to be illustrative and are not meant to limit the scope of the present disclosure. The following examples are meant to illustrate the disclosed embodiments and are not to be construed as limiting thereof. Other compounds than those described below can be prepared by following the reaction schemes below or appropriate variations or modifications thereof.

[0342] Example 1: Synthesis of 2-chlorobenzylamino-2-imidazoline hydroiodide

[0343] To a stirred solution of 2-methylthio-2-imidazoline hydroiodide (244 mg, 1.00 mMol) in anhydrous dioxane (2.0 mL) was added 2-chlorobenzylamine (141 mg, 1.0 mMol). The reaction mixture was stirred at 70 °C under argon for 90 minutes. The solution was cooled to room temperature, filtered over a sintered funnel, washed with cold dioxane (2 mL) and dried in vacuo. Compound 4»HI (R2= 2-chlorobenzyl) was obtained as a white solid (242 mg, 72%) and used without further purification.

[0344] Example 2: Synthesis of 2-chlorobenzylamino-2-imidazoline Synthesis

[0345] To a stirred solution of 4-oxo-3-piperidinecarboxylic acid methyl-l-benzyl ester hydrochloride (5.7 g, 20 mMol) in ethyl acetate (50 mL) at 7°C was added triethylamine (6 mL). The reaction mixture was stirred at 7°C under argon atmosphere for 30 minutes. The reaction mixture was extracted with ethyl acetate (2 x 50 mL) and washed with water (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated. The resulting free base residue (5, R1= benzyl) was a thick oil and used in the next reaction without any further purification. MS (ESI) 248 (M+H).

[0346] Example 3: Synthesis of 4-oxo-3-piperidinecarboxylic acid methyl-1- benzyl ester (Compound (6)).

[0347] To a stirred solution of 4-oxo-3-piperidinecarboxylic acid methyl-l-benzyl ester hydrochloride (5.7 g, 20 mMol) in ethyl acetate (50 mL) at 7°C was added triethylamine (6 mL). The reaction mixture was stirred at 7°C under argon atmosphere for 30 minutes. The reaction mixture was extracted with ethyl acetate (2 x 50 mL) and washed with water (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated. The resulting free base residue (5, R1= benzyl) was a thick oil and used in the next reaction without any further purification. MS (ESI) 248 (M+H).

[0348] Example 4: Synthesis of ONC202 (Compound (14))

[0349] To a stirred solution of 4-oxo-3-piperidinecarboxylic acid methyl-l-benzyl ester hydrochloride (5.7 g, 20 mMol) in ethyl acetate (50 mL) at 7°C was added triethylamine (6 mL). The reaction mixture was stirred at 7°C under argon atmosphere for 30 minutes. The reaction mixture was extracted with ethyl acetate (2 x 50 mL) and washed with water (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated. The resulting free base residue (5, R1= benzyl) was a thick oil and used in the next reaction without any further purification. MS (ESI) 248 (M+H).

[0350] Various analogs were prepared using the same procedure starting from different benzylamines, for example ONC203, 204, 205, 206, 912, 210, 211, 212, 213, 214, 217, 218, 219, 220, 221, 222, 223, 224, 225 and 226.

[0351] Various analogs were prepared using the same procedure starting from different benzylamines, for example ONC203, 204, 205, 206, 912, 210, 211, 212, 213, 214, 217, 218, 219, 220, 221, 222, 223, 224, 225 and 226.

[0351] Various analogs were prepared using the same procedure starting from different benzylamines, for example ONC203, 204, 205, 206, 912, 210, 211, 212, 213, 214, 217, 218, 219, 220, 221, 222, 223, 224, 225 and 226.Example 5: Synthesis of ONC207 (Compound (19))

[0352] To a suspension of 60% sodium hydride (3.5 g, 88 mMol) in dry toluene (50 mL) was added dimethyl carbonate (4.32 g, 48.0 mMol) dropwise over 0.5 hours at room temperature under nitrogen. After the addition of a few drops of methanol, 1-tert-butoxycarbonyl-4-piperidinone (4.8 g, 24 mMol) dissolved in dry toluene (20 mL) was added dropwise to the reaction mixture over 1 hour with stirring at 80 °C. The reaction mixture was stirred at the same temperature for 3 hours, then cooled to 0 °C (ice bath) and adjusted to pH 6-6.5 with acetic acid. The resulting cold mixture was diluted with water (10 mL) and adjusted to pH 8 with 5% sodium hydroxide solution. The toluene layer was separated and the aqueous layer was extracted with toluene (20 mL). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The compound was dried in vacuum to yield 1-tert-butoxycarbonyl-4-oxo-3-piperidinecarboxylic acid methyl ester (5.0 g, 80%). The resulting compound was carried on to the next reaction without any further purification.

[0353] To a solution of 2-chlorobenzylamino-2-imidazoline (190 mg, 1 mMol), 1-tert-butoxycarbonyl-4-oxo-3-piperidinecarboxylic acid methyl ester (315 mg, 1.1 mMol) in 1-butanol (2 mL) was added PPTS (10 mg) and the mixture was stirred at room temperature for 48 hours. After this time, the reaction mixture was refluxed at 125 °C to 130 °C for 2 hours. The solvent was removed in vacuum, extracted with ethyl acetate (10 mL), washed with saturated sodium bicarbonate solution (2 x 10 mL) and water (10 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated. The crude free base was cleaved with 10% trifluoroacetic acid in dichloromethane, purified by RP HPLC (10% to 40% acetonitrile / water) to yield ONC907 (262 mg, 50%) as a TFA salt as a white solid, MS (ESI) 297 (M+H).

[0354] Example 6: Synthesis of ONC209 (Compound (21))

[0355] A mixture of ONC907 (100 mg, 0.2 mMol), phenethyl bromide (55.0 mg, 0.28 mMol) and potassium carbonate (150 mg, 1.0 mMol) in N,N-dimethylformamide (3 mL) was heated to 70 °C for 12 hours. The solvent was removed in vacuum, extracted with ethyl acetate (10 mL) and washed with water (5 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated. The crude free base was purified by RP HPLC (10% to 40% acetonitrile / water) to yield ONC209 (62 mg, 50%) as a TFA salt as a white solid, MS (ESI) 401 (M+H).

[0356] ONC 215 and 214 were produced using the same procedure starting from different halides. Compounds 227, 228, 229, 230, 231, 232, 233, 234, 235 and 236 were prepared starting from different benzylamines using a similar procedure to that of Examples 1 and 5. The intermediate compounds with R1 as H were then treated with different halides as described above.

[0357] Compound ONC 216 was prepared from ONC 215 by treatment with TFA.

[0358] Compound (72) was prepared by reacting a precursor NH compound prepared similarly to Example 5 and treating it with Oxone.

[0359] Example 7: Synthesis of ONC208 (Compound (20))

[0360] To a solution of 2-chlorobenzylamino-2-imidazoline (190.0 mg, 1.0 mmol), 4-oxo-3-piperidinecarboxylic acid methyl ester (185.0 mg, 1.0 mMol) in 1-butanol (2 mL) was added PPTS (10 mg) and the mixture was stirred at room temperature for 48 hours. After that, the reaction mixture was refluxed at 125-130 °C for 2 hours. The solvent was removed in vacuo, extracted with ethyl acetate (10 mL), washed with saturated sodium bicarbonate solution (2 x 10 mL) and water (10 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated. The crude free base was purified by HPLC 10%-40% acetonitrile and water to give ONC 908 (270.0 mg, 50%) as a TFA salt, white solid, MS (ESI) 311 (M+H).

[0361] Example 8: Synthesis of ONC201 (Compound (1))

[0362] To a stirred 800 mL saturated NaHC03in a 2 L round bottom flask was added compound (3) (239.7 g, 0.845 mol, 1.6 eq) in portions. To the resulting mixture was added n-butanol (500 mL) and stirred for 30 minutes before transferring to a separatory funnel. The organic phase containing compound (4) was isolated and transferred to a 2 L three necked round bottom flask equipped with mechanical stirring, N2inlet, thermocouple, condenser and Dean-Stark trap. To the contents of the flask was added compound (5) (100 g, 0.528 mol, 1 eq) and p-toluenesulfonic acid pyridinium (PPTS) (6.63 gm 0.026 mol, 5 mol%). The resulting mixture was heated to reflux for 6 hours. Water was removed from the reaction mixture into the Dean-Stark trap as needed. The reflux temperature was increased from 93 °C to 118 °C. The progress of the reaction was monitored by HPLC. The reaction was stopped when the peak area of compound (1) remained constant with time on HPLC.

[0363] Example 9: Synthesis of ONC201 di-salt (Compound (1) 2HC1)

[0364] The reaction mixture of Example 8 was not isolated, washed with water (500 mL) and diluted with methyl tert-butyl ether (MTBE) (800 mL). The organic phase was washed with water (500 mL x 2) and transferred to a 3 L three necked round bottom flask equipped with mechanical stirring, N2inlet, thermocouple, condenser and Dean-Stark trap. While stirring the reaction mixture, a solution of 1 N HC1 in dioxane-METBE (4 N HC1 in dioxane: 300 mL, 1.2 mol, 2.27 eq; MTBE: 1200 mL) was added dropwise until no more solids precipitated from the reaction mixture upon addition of HC1. The reaction mixture was heated to reflux at 60-65 °C for 2 hours. Water was removed into the Dean-Stark trap as needed. After cooling to room temperature, the solid precipitate was filtered through a sintered glass funnel and washed with n-butanol-MTBE (1 :2, 600 mL) and MTBE (600 ...

Claims

1. Use of a compound of formula (1) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating central nervous system cancer in a subject, The central nervous system cancer is recurrent glioblastoma with a histone H3 mutation.

2. The use according to claim 1, wherein the histone H3 mutation is H3.3 K27M.

3. The use according to claim 1, wherein the cancer has an epigenetically silenced unmethylated O(6)-methylguanine-DNA methyltransferase (MGMT) gene.

4. The use according to any one of claims 1 to 3, wherein the subject is a human.

5. The use according to any one of claims 1 to 3, wherein the pharmaceutically acceptable salt is dihydrochloride.

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