Modulation of g-protein coupled receptors (GPCRS) by imipridones

Abstract

To provide an agent for reducing tumor size relative to baseline of a central nervous system cancer.SOLUTION: The medicine contains a compound represented by structural formula (I) or a pharmaceutically acceptable salt thereof.SELECTED DRAWING: None

Description

[Background technology]

[0001] Human cells possess a variety of receptors on their surfaces. G protein-coupled receptors (GPCRs) form one of the largest protein families of transmembrane receptors. The human genome contains approximately 30,000 genes, of which 1,000 encode GPCRs. GPCRs have been classified into five classes. The first class is the rhodopsin receptor family, or "class A GPCRs," which includes 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 a large number of drug target receptors. The second class is the secretin receptor family, which has binding domains for peptide hormones. Receptors in this family are involved in homeostasis and have emerged as important targets for drug development. The third class is the adhesion receptor family, which is characterized by GPCR proteolytic sites (GPSs). This family of GPCRs has a variety of N-terminal segments, and few ligands have been identified, preventing the development of drugs targeting this family. The fourth class is the glutamate receptor family, with 22 GPCR members identified to date. Relatively little is known about the specificity of each protein. The final class is the Frizzled / Taste2 family, which includes 10 Frizzled receptors for which Wnt glycoproteins serve as ligands, five ligand-free SMO (smoothened) receptors, and 25 Taste2 receptors required for the perception of various tastes. Receptors, including GPCRs, are also classified based on the identification of their exogenous ligands. Receptors bind either known endogenous compounds or are classified as orphan receptors, for which no endogenous ligand has yet been identified.

[0002] GPCRs are found in a wide range of tissues and cell types and are involved in many different physiological mechanisms. GPCRs react with a variety of ligands, including hormones such as thyroid-stimulating hormone (TSH), adrenocorticotropic hormone, glucagon, and vasopressin; amines such as 5-HT, acetylcholine (muscarinic AchR) and histamine; lipids and amino acids such as LPA and S1P; and Ca. 2+ GPCRs are activated by signaling factors such as ATP, nucleic acids, peptides, and light. The wide distribution and diversity of roles played by GPCRs provides evidence of their importance in various pathological disorders. Indeed, GPCRs are involved in a variety of diseases, including bronchoconstriction, hypertension, diabetes, inflammation, cell death, hormonal disorders, cancer, neurotransmitter disorders, and behavioral disorders. This makes GPCRs an important area of ​​focus for drug development. Currently, approximately 360 GPCRs are available for drug development. Of these, 46 are already in drug development. An estimated 150 orphan GPCRs (oGPCRs) exist. In the field of drug development, cell membrane receptors serve as the selective site of drug action, contributing to 50% of all drug targets. Drugs modulating GPCR activity account for 30% of the top 100 most frequently used drugs ($40 billion, or 9% of the total pharmaceutical market). Therefore, GPCRs are among the most important targets for new drug development.

[0003] GPCRs share common structural features. They contain seven hydrophobic transmembrane domains, each consisting of 20–30 amino acids and connected by hydrophilic amino acid sequences of various lengths. Receptors have an extracellular N-terminus and a C-terminus located within the cytoplasm. GTP-binding proteins (G proteins) act as mediators, transmitting signals generated by GPCR binding to hormones or other chemical ligands to intracellular effectors. After ligand binding, a conformational change in the intracellular domain of the GPCR allows the receptor to interact with the G protein, which then activates intracellular signaling factors such as adenylate cyclase, phospholipase C, or ion channels. In this system, binding of a single ligand to a GPCR initiates a signaling cascade involving the activation of multiple second messengers. Cells use this mechanism to sense changes in the extracellular environment and respond appropriately to those changes. Overall, endogenous ligands activate receptors, simultaneously triggering a conformational change that allows the receptor to associate with the G protein. Recent studies on protein-protein interactions have revealed that GPCRs associate with various proteins, such as GRK or SH2 (Src Homology 2) domain-containing proteins and the adaptor Grb2, as well as G proteins, to mediate signal transduction.

[0004] Under normal conditions, signal transduction ultimately results in cellular activation or inhibition. In a physiological environment, GPCRs exist in an equilibrium state between inactive and active states within the cell membrane. Inactive receptors cannot coordinate intracellular signaling pathways to produce biological responses. Receptors exert biological responses via signal transduction pathways (mediated by G proteins) only when they undergo a conformational change to their active form. Receptors can be stabilized in their active form by endogenous ligands or compounds such as drugs. Therefore, functional studies, such as cloning such gene families and identifying their novel ligands, are synonymous with the development of new drug candidates, namely, siRNAs, antibodies, polypeptides, effectors, inhibitors, agonists, and antagonists.

[0005] Most biological processes, including development, differentiation, homeostasis, response to stimuli, cell cycle control, aging, and apoptosis, are driven by the selective expression of specific genes within cells. This also applies to the intracellular mechanisms associated with disease. Specifically, pathological phenomena such as carcinogenesis are caused by genetic mutations that ultimately lead to altered gene expression.

[0006] ONC201 (7-benzyl-4-(2-methylbenzyl)-1,2,6,7,8,9-hexahydroimidazo[1,2-a]pyrido[3,4-e]pyrimidin-5(1H)-one) is an established member of a class of anticancer compounds known as imipridones and is currently in Phase II clinical trials for multiple advanced cancers. Since its discovery as a p53-independent inducer of TRAIL gene transcription, preclinical studies have demonstrated its antiproliferative and proapoptotic effects in a broad range of tumor cells but not in normal cells. The mechanism of action of ONC201 involves PERK-independent activation of the integrated stress response, which leads to tumor DR5 upregulation, coupled with Akt / ERK dual inactivation and subsequent Foxo3a activation, which leads to upregulation of the death ligand TRAIL. In animal models, ONC201 is orally active with infrequent administration, produces sustained pharmacodynamic effects, and exhibits no genotoxicity. First-in-human ONC201 clinical trials in advanced, aggressive, treatment-resistant solid tumors confirmed that ONC201 was well tolerated. Collectively, the imipridone family, which includes ONC201 and its chemical analogues, represents a new class of therapeutic agents. Summary of the Invention [Means for solving the problem]

[0007] In one aspect, the present invention provides a compound represented by the following structural formula (10):

[0008] [ka]

[0009] wherein R1 and R2 are independently selected from H, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, and acyl radicals. In one embodiment, when R1 is CH2Ph, R2 is not CH2-(2-CH3-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2-CH3-Ph) (i.e., ONC201). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2,4-diF-Ph) (i.e., ONC206). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(4-CF3-Ph) (i.e., ONC212). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(3,4-diF-Ph) (i.e., ONC213). In one embodiment, R1 is CH2(3,4-di-Cl-Ph) and R2 is CH2-(4-CF3-Ph) (i.e., ONC234). In one embodiment, R1 is CH2-3-thienyl and R2 is CH2-(4-CF3-Ph) (i.e., ONC236).

[0010] In another aspect, the present invention provides a method for treating or preventing a disease, disorder, or condition in a subject in need thereof, comprising administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a compound of structural formula (10) or an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is selected from the group consisting of ONC201, ONC206, ONC212, ONC213, ONC234, and ONC236. In one embodiment, the subject has or is at risk of having cancer. In one 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, Ewing's sarcoma, pancreatic cancer, prostate cancer, endometrial cancer, a hematological malignancy, a bone cancer, and a lymphatic tumor. In one embodiment, the cancer is selected from a meningioma, an ependymoma, a glioma, a neuroblastoma, or a diffuse intrinsic pontine glioma. In one embodiment, the cancer is an acute leukemia selected from acute lymphocytic leukemia, acute myeloid leukemia, myelodysplastic syndrome, or myeloproliferative disorder. In one embodiment, the cancer has a histone H3 mutation (e.g., mutation H3.3 K27M) or an epigenetically silenced unmethylated O(6)-methylguanine-DNA methyltransferase (MGMT) gene. In one embodiment, the subject has or is at risk of having a psychiatric disorder. In one embodiment, the psychiatric disorder is selected from 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 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 Staphylococcus infection. In one embodiment, the Staphylococcus infection is a S. aureus infection (e.g., a methicillin-resistant S. aureus (MRSA) infection).

[0011] In another aspect, the present invention provides a method for treating or preventing a disease, disorder, or condition in a subject requiring selective modulation of the activity 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 bitopic 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 structural formula (10) or an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has or is at risk of having 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 Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, or an Enterobacter species. In one embodiment, the bacterial infection is a Staphylococcus infection. In one embodiment, the Staphylococcus infection is a S. aureus infection (e.g., a methicillin-resistant S. aureus (MRSA) infection). In one embodiment, the treatment regimen comprises administering an effective amount of a therapeutic agent, such as a compound of structural 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.

[0012] In another aspect, the present invention provides a method for treating or preventing a disease, disorder, or condition in a subject requiring selective modulation of the activity of a dopamine receptor or a member of a 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 structural formula (10) or an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has or is at risk of having 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 a bacterial infection 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 Staphylococcus infection. In one embodiment, the Staphylococcus infection is a S. aureus infection (e.g., a methicillin-resistant S. aureus (MRSA) infection). In one embodiment, the treatment regimen comprises administering an effective amount of a therapeutic agent, e.g., a compound of structural formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is of the D2-like family of dopamine receptors.

[0013] In another aspect, the present invention provides a method for treating or preventing liver fibrosis or regenerating liver tissue, 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.

[0014] In another aspect, the present invention provides a method of stimulating the immune system (e.g., activating NK cells) in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a compound of structural 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), wherein the compound is administered as an adjuvant.

[0015] In another aspect, the present invention provides a method for determining whether a subject having a pathological condition is likely to respond to a treatment regimen described herein. In one embodiment, the method includes (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 with that of a predetermined standard; and (iv) determining whether the subject is likely to respond to the treatment regimen based on the level measured in the sample relative to that of the predetermined standard. In one embodiment, the subject has or is at risk of having 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 infectious disease. In one embodiment, the treatment regimen further includes administering an effective amount of a therapeutic agent, such as a compound of structural formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is of the D2-like family of dopamine receptors. 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.

[0016] In another aspect, the present invention provides a method for evaluating the effectiveness of a treatment regimen described herein, for monitoring, or for predicting the prognosis of a subject with a condition. In one embodiment, the method includes: (i) collecting 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 a predetermined standard; and (iv) determining the prognosis or whether the subject will respond to the treatment regimen based on the level measured in the sample relative to the predetermined standard. In one embodiment, the method includes: (i) collecting 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 measured copy number or mutation found in the sample with a predetermined standard; and (iv) determining whether the subject will respond to the treatment regimen based on the measured copy number or mutation found in the sample relative to the predetermined standard. In one embodiment, the subject has cancer or is at risk of having cancer. In one embodiment, the subject has a psychiatric disorder 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 comprises administering an effective amount of a therapeutic agent, such as a compound of structural 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 family of dopamine receptors. 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.

[0017] In another aspect, the present invention provides a method for screening potential therapeutic agents for a disease state. In one embodiment, the method comprises: (i) contacting at least one G protein-coupled receptor (GPCR) with a test molecule that is a potential therapeutic agent for the disease state; (ii) measuring the binding affinity, interaction, or GPCR 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, modulation of the GPCR or modulation of GPCR signaling by the test molecule that is equal to or greater than the threshold is indicative of a therapeutic agent for the disease state. In one embodiment, the disease state is cancer. In one embodiment, the predetermined threshold is modulation of the GPCR or modulation of GPCR signaling by a therapeutic agent, such as a compound of structural 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.

[0018] In another aspect, the present invention provides a method for screening for potential therapeutic agents for a disease state. In one embodiment, the method comprises: (i) contacting at least one dopamine receptor with a test molecule suspected to be a therapeutic agent for the disease state; (ii) measuring the binding affinity, interaction, or signaling of the test molecule with the at least one dopamine receptor; and (iii) comparing the binding affinity or interaction of the test molecule to a predetermined threshold. In one embodiment, modulation of the dopamine receptor by the test molecule at or above the threshold is indicative of a therapeutic agent for the disease state. In one embodiment, the disease state is cancer. In one embodiment, the dopamine receptor is a member of the D2-like family of dopamine receptors. In one embodiment, the predetermined threshold is modulation of the dopamine receptor or dopamine receptor signaling by a therapeutic agent, such as a compound of structural formula (10) or a pharmaceutically acceptable salt thereof or an analog thereof.

[0019] In another aspect, the present invention provides a method for screening potential therapeutic agents for a disease state. In one embodiment using a processor, the method includes: (i) using a computational docking method to model the binding or interaction (if any) of one or more three-dimensional structures (conformations) of a test molecule that is a potential therapeutic agent for the disease state with a three-dimensional structure or model of at least one dopamine receptor; (ii) using a computational method to estimate the binding affinity or interaction between the test molecule structure and the structure or model of at least one dopamine receptor; and (iii) using a computational method to compare the binding affinity or interaction of the test molecule with a predetermined threshold, wherein modulation of the dopamine receptor by the test molecule at or above the threshold is indicative of a therapeutic agent for the disease state. In one embodiment, the disease state is cancer. In one embodiment, the dopamine receptor is a member of the D2-like family of dopamine receptors.

[0020] In another aspect, the present invention provides a method of treating a subject having a pathological condition. In one embodiment, the method comprises administering an effective amount of a therapeutic agent that targets 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 a receptor antagonist. In one embodiment, the therapeutic agent is a receptor agonist. In one embodiment, the therapeutic agent is a competitive inhibitor of a receptor for dopamine. In one embodiment, the therapeutic agent is a non-competitive inhibitor of a receptor for dopamine. In one embodiment, the therapeutic agent is selective for the D2-like family of dopamine receptors relative to the D1-like family of dopamine receptors. In one embodiment, the subject has or is at risk of having 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 infectious disease. In one embodiment, the dopamine receptor is a member of the D2-like family of dopamine receptors. 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., a chimerized or humanized monoclonal antibody), a polyclonal antibody (e.g., a chimerized or humanized polyclonal antibody), or a bispecific antibody. In one embodiment, the therapeutic agent is a drug or active agent, such as an anti-cancer agent, conjugated to the antibody. In one embodiment, the therapeutic agent is a radioactive substance-conjugated antibody or a small molecule-conjugated antibody. In one embodiment, the therapeutic agent is a vector expressing a recombinant antibody against a dopamine receptor or a GPCR. In one embodiment, the therapeutic agent is a fusion protein or peptide targeting a dopamine receptor or a GPCR.In one embodiment, the therapeutic agent is an siRNA, shRNA, or antisense oligonucleotide that targets a dopamine receptor or GPCR. In one embodiment, the dopamine receptor or GPCR is targeted by CRISPR interference.

[0021] In another aspect, the present invention provides a method for treating a subject having a pathological condition and assessing the effectiveness of the treatment. In one embodiment, the method comprises (i) treating the subject with a treatment method described herein and (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 administering an effective amount of a therapeutic agent, such as a compound of structural formula (10) or a pharmaceutically acceptable salt or analog thereof. In one embodiment, the dose of the administered therapeutic agent, the frequency of administration of the compound (e.g., a compound of structural formula (10)), or both, is selected or adjusted based on the measured level of gene expression or gene copy number or the mutations found.

[0022] The foregoing summary, as well as the following detailed description of embodiments of the invention, will be better understood when read in conjunction with the appended drawings, it being understood, however, that the invention is not limited to the precise arrangements and instrumentalities shown. [Brief explanation of the drawings]

[0023] [Figure 1] FIG. 1 shows the antagonism of dopamine receptors (DRD1, DRD2S, DRD2L, DRD3, DRD4 and DRD5) by ONC201. [Figure 2] Figure 1 shows soluble prolactin detected in peripheral blood of patients with advanced solid tumors by ELISA at baseline and after a single dose of ONC201 (125-625 mg PO). Post-treatment sampling time points include 6 hours, 1 day, 2 days, 7 days, and 21 days after treatment. [Figure 3]Figure 1 shows the sensitivity of tumor types in the Genomic of Drug Sensitivity in Cancer program (GDSC) cell line collection. Mean sensitivity was determined by the mean IC50 value estimated from cell viability assays performed 72 hours after treatment. Numbers above the bars represent the number of cell lines per tumor type. [Figure 4A] FIG. 1 shows that ONC201 is a selective DRD2 antagonist, demonstrating agonism of orphan or known GPCRs or antagonism of known GPCRs using an arrestin recruitment reporter assay (10 μM ONC201). [Figure 4B] FIG. 1 shows that ONC201 is a selective DRD2 antagonist. Antagonism of ligand-stimulated dopamine receptors by ONC201 is shown using an arrestin recruitment reporter assay. [Figure 4C] FIG. 1 shows that ONC201 is a selective DRD2 antagonist. Shield analysis of DRD2L antagonism by ONC201 using an arrestin recruitment reporter is shown. [Figure 4D] FIG. 1 shows that ONC201 is a selective DRD2 antagonist. Shield analysis of DRD2L antagonism by ONC201 using a cAMP-regulated reporter. [Figure 5A] FIG. 1 shows the highly specificity of ONC201 antagonism of DRD2 among GPCRs and other cancer therapeutic targets. Antagonism of GPCRs is shown using an arrestin recruitment reporter assay (10 μM ONC201). [Figure 5B] FIG. 10 shows the high specificity of ONC201 antagonism of DRD2 among GPCRs and other cancer therapeutic targets. Competition of ONC201-mediated antagonism of DRD2L by dopamine in an arrestin recruitment reporter. [Figure 5C] FIG. 10 shows the high specificity of ONC201 antagonism of DRD2 among GPCRs and other cancer therapeutic targets. Competition of ONC201-mediated antagonism of DRD2L by dopamine on a cAMP-regulated reporter. [Figure 5D] Figure 1 shows the highly specificity of ONC201 antagonism of DRD2 among GPCRs and other cancer therapeutic targets. Nuclear hormone receptor antagonism or agonism by ONC201 (2 μM or 20 μM) using a nuclear translocation reporter assay. [Figure 5E] FIG. 1 shows the highly specificity of ONC201 antagonism of DRD2 among GPCRs and other cancer therapeutic targets, showing in vitro inhibition of kinase enzyme activity by ONC201 (1 μM). [Figure 5F] FIG. 1 shows the highly specificity of ONC201 antagonism of DRD2 among GPCRs and other cancer therapeutic targets. The figure shows the DRD2L antagonism activity of ONC201 or the non-biologically active linear isomer of ONC201 using an arrestin recruitment reporter assay. [Figure 6] Figure 1 shows that GBM cell lines with higher DRD2 expression respond better to ONC201. (A) Inhibition of NCI60 GBM cell line as a function of ONC201 concentration. (B) Log ONC201 GI50(M) vs. DRD2 expression for each GBM cell line. R2=0.8707. [Figure 7] FIG. 1 shows that ONC201 exhibits superior selectivity for DRD2 among GPCRs compared to other DRD2 antagonists such as risperidone. [Figure 8] FIG. 1 shows that ONC201 is more selective for tumor cells than the antipsychotic DRD2 antagonist thioridazine. [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 examined with 0.1 nM and 1 nM dopamine for ONC201 concentrations ranging from 100 pM to 100 μM. 100 μM ONC201 completely inhibited DRD2 dopamine-induced calcium flux but had no effect on the control GPCR. [Figure 10]Figure 1 shows a comparison of DRD2 inhibitors. The inhibitors spiperone (squares), domperidone (triangles), and ONC201 (circles) were used at various concentrations to examine DRD2-specific calcium flux inhibition at 1 nM dopamine. Data from individual assays were normalized to the inhibitor-free value (shown as 10-11 M) as 100% activity. [Figure 11] Figure 1. Identification of DRD2 residues critical for dopamine-induced calcium flux. (A) Dopamine-induced calcium flux at 1 nM dopamine was assayed across the DRD2 alanine-scan library as described above. Data represent the mean of triplicate experiments. Mutant clones were considered calcium flux-deficient if they exhibited flux values ​​below two standard deviations (AV-2SD) of the mean calcium flux value across the entire library. (B) The locations of the 28 identified mutant residues are shown (green spheres) on the DRD3 crystal structure (PDB id 3PBL; Chien, EY et al. (2010) Science 330:1091-5). The D2R / D3R antagonist eticlopride is shown in cyan. [Figure 12] Figure 1. Identification of DRD2 residues critical for ONC201 inhibition of dopamine-induced calcium flux. (A) Dopamine-induced calcium flux at 1 nM dopamine was assayed across the DRD2 alanine-scan library as described above, except in the presence of 100 μM ONC201. Data represent the mean of triplicate experiments normalized to the wild-type DRD2 flux value (%WT). Mutant clones were considered critical for ONC201 inhibition if they exhibited flux values ​​greater than two standard deviations above the mean calcium flux value for the entire library (AV + 2SD). (B) The locations of the eight identified mutant residues are shown on the DRD3 crystal structure (red spheres). [Figure 13] FIG. 1 shows that the reference compound (+)-butaclamol and the test compound ONC201 dihydrochloride competed well for [3H]methylspiperone, with IC50 values ​​of 2.5 nM and 21 μM, respectively. [Figure 14] FIG. 1 shows the binding rate curves for determining Kon and Koff between ONC201 dihydrochloride and the DRD2S receptor. [Figure 15] Figure 1 shows compound activity using biosensor assays for selected GPCRs and orphan GPCRs. Compounds were tested in antagonist and agonist modes using biosensor assays for the desired GPCRs and orphan GPCRs. 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 EC80 ligand and vehicle. The following EC80 concentrations were used: CCR4 arrestin: 0.0078 μM CCL22; CHRM2 arrestin: 26 μM acetylcholine; and MC4R arrestin: 0.0026 μM melanotan II. [Figure 16] 1 shows that ONC206 and ONC212 exhibited anti-cancer effects on various tumor types in the NCI60 cancer cell line panel. ONC203 is an inactive negative control. [Figure 17] ONC206 is an imipridone with improved DRD2 antagonism. ONC206, an analog of ONC201, exhibits superior antagonism at the D2-like dopamine receptor family compared to other antipsychotics such as haloperidol, while retaining highly selective antagonism at D2-like dopamine receptors. [Figure 18] FIG. 1 shows that bone cancer is more responsive to ONC206 than to ONC201. [Figure 19] FIG. 1 shows that Ewing's sarcoma is the most responsive bone cancer subtype to ONC206. [Figure 20] Figure 1 shows that the anti-cancer effect of ONC206 is in the nanomolar range in 14 of 16 Ewing's sarcoma cell lines. ONC206 showed superior efficacy to ONC201 in all cell lines. [Figure 21]1 shows that the imipridone ONC212 targets orphan GPCRs. ONC212 is a highly selective agonist of the orphan GPCR tumor suppressor GPR132 and does not bind to DRD2. [Figure 22] FIG. 1 shows that ONC212 induced cell death in cancer cells (HCT116) but not in normal cells (MRC5) at nanomolar concentrations. [Figure 23] FIG. 1 shows that ONC212 induces the integrated stress response and inhibits Akt / ERK phosphorylation at nanomolar concentrations at earlier time points than ONC201. [Figure 24] FIG. 1 shows that ONC212 exhibits oral and IP anticancer efficacy in xenograft mouse models of colorectal and breast cancer. [Figure 25] FIG. 1 shows that leukemia is more responsive to ONC212 than to ONC201. [Figure 26] FIG. 1 shows that ONC212 exhibits anti-cancer efficacy (and superior efficacy to ONC201) in the nanomolar concentration range against 55 leukemia cell lines of any subtype. [Figure 27-1] Figure 1 shows that GPCRs were stimulated or antagonized (>50%) by the nine imipridones tested. Imipridones selectively target rhodopsin-like class A GPCRs. [Figure 27-2] Figure 1 shows that GPCRs were stimulated or antagonized (>50%) by the nine imipridones tested. Imipridones selectively target rhodopsin-like class A GPCRs. [Figure 28] Figure 16 shows a case study of a subject with recurrent glioblastoma (Example 16). (A) Tumor size (%) of the subject's total tumor burden relative to baseline. One cycle is 3 weeks. (B) Contrast-enhanced MRI scans of one of two malignant lesions at baseline, 21 weeks, 27 weeks, and 36 weeks after starting ONC201. [Figure 29A]FIG. 1 shows that ONC212 exhibits anti-cancer activity in acute myeloid leukemia (AML) cell lines, showing a comparison of cell viability of MV411 AML cells treated with ONC212 or cytarabine. [Figure 29B] FIG. 1 shows that ONC212 exhibits anti-cancer activity in acute myeloid leukemia (AML) cell lines, comparing cell viability of MOLM14 cells, MV411 AML cells, MRC5 lung fibroblasts, and Hs27a bone marrow cells treated with ONC212. [Figure 29C] FIG. 1 shows that ONC212 exhibits anti-cancer activity in acute myeloid leukemia (AML) cell lines, showing the cell viability of MOLM14 and MV411 AML cells treated with ONC212 (250 nM) for 4 hours, 8 hours, 24 hours, 48 ​​hours, 72 hours, and 96 hours. [Figure 30A] Figure 1 shows the effect of ONC212 in an ONC201-resistant AML xenograft model (MV411 AML cells (5x106) implanted subcutaneously in the flanks of athymic nude mice). ONC212 and ONC201 were administered orally (PO) as indicated. Tumor volumes (n=10) were measured on the indicated days. * indicates p<0.05 vs. vehicle. [Figure 30B] Figure 1 shows the effect of ONC212 in an ONC201-resistant AML xenograft model (MV411 AML cells (5x106) implanted subcutaneously in the flanks of athymic nude mice). ONC212 and ONC201 were administered orally (PO) as indicated. Tumor volumes (n=10) were measured on the indicated days. * indicates p<0.05 vs. vehicle. [Figure 30C] Figure 1 shows the effect of ONC212 in an ONC201-resistant AML xenograft model (MV411 AML cells (5 x 106) implanted subcutaneously in the flanks of athymic nude mice). ONC212 and ONC201 were administered orally (PO) as indicated. Body weights (n=10) were measured on the indicated days. [Figure 31]Effect of ONC206 on a Ewing sarcoma xenograft model (MHH-ES-1 Ewing sarcoma cells (5x10) implanted subcutaneously in the flanks of athymic nude mice). ONC206 (PO) and methotrexate (IV) were administered on days 1 and 13 as indicated. Tumor volume (A) and body weight (B) (n=4) were measured on the days indicated. [Figure 32] FIG. 1 shows the GPCR profile of ONC213 (10 μM) using a β-arrestin recruitment reporter assay. [Figure 33] FIG. 1 shows that ONC213 exhibited anticancer efficacy against HCT116 / RPMI8226 cancer cells in vitro comparable to ONC212, but had lower in vitro toxicity against normal cells than ONC212. [Figure 34] FIG. 1 shows the GPCR profile of ONC237 (10 μM) using a β-arrestin recruitment reporter assay. [Figure 35] FIG. 1 shows the GPCR profile of ONC236 (10 μM) using a β-arrestin recruitment reporter assay. [Figure 36] FIG. 1 shows the GPCR profile of ONC234 (10 μM) using a β-arrestin recruitment reporter assay. [Figure 37] FIG. 1 shows the GPCR profile of ONC201 linear isomer (TIC-10) (10 μM) using a β-arrestin recruitment reporter assay. [Figure 38] FIG. 1 shows the number of GPCR hits for several imipridones. DETAILED DESCRIPTION OF THE INVENTION

[0024] Scientific and technical terms used herein shall have meanings that are commonly understood by those of ordinary skill in the art.Such terms are used in, for example, J. Sambrook and DWRussell, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press; 3rd Ed., 2001; FM Ausubel, Ed., Short Protocols in Molecular Biology, Current Protocols; 5th Ed., 2002; MMCox,Lehninger Principles of Biochemistry,4th Ed.,WHFreeman & Company,2004;Engelke,DR,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. Wintersten, R. Behringer, Manipulating the Mouse Embryo: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory Press; December 15, 2002, ISBN-10: 0879695919; Kursad Turksen (Ed.), Embryonic stem cells: methods and protocols in 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 above documents are incorporated herein by reference in their entirety.

[0025] The term "substituted" means replacing any one or more hydrogens on the specified atom with a group selected from the indicated group, provided that the substitution does not exceed the normal valence of the specified atom and results in a stable compound. When the substituent is keto (i.e., =0), two hydrogens on the atom are replaced. Keto substituents do not occur 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).

[0026] Variables (e.g., R 4 ) occurs more than one time, its definition on each occurrence is independent of its definition at every other occurrence. Thus, for example, a group may contain 0 to 3 R 4 When a moiety is indicated to be substituted, the group may optionally be substituted with up to three R 4 may be substituted with the moiety, and each occurrence of R 4 is R 4 Combinations of substituents and / or variables are permissible, but only if such combinations result in stable compounds.

[0027] When an atom or chemical moiety is followed by a subscripted numerical range (e.g., C 1~6 ), it will be understood that each value within that range and all intermediate ranges are encompassed. For example, "C 1~6 "Alkyl" is meant to include alkyl groups having 1, 2, 3, 4, 5, 6, 1-6, 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.

[0028] The term "alkyl" includes both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, C 1~6Alkyl is intended to include C1, C2, C3, C4, C5, and C6 alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, s-pentyl, neopentyl, and n-hexyl. In certain cases, a straight chain or branched chain alkyl has 6 or fewer carbon atoms in its backbone (e.g., C1-C6 for straight chain, C3-C6 for branched chain), and in other cases, a straight chain or branched chain alkyl has 4 or fewer carbon atoms. Likewise, cycloalkyls have from 3 to 8 carbon atoms in their ring structure, and in other cases, cycloalkyls have 5 or 6 carbons in the ring structure. Most preferred are C 1~6 Alkyl, in particular ethyl, methyl, isopropyl, isobutyl, n-pentyl, n-hexyl and cyclopropylmethyl.

[0029] The term "substituted alkyl" refers to alkyl, as defined above, substituted with one, two, or three substituents selected from halogen, -OH, alkoxy, -NH, -N(CH), -C(=O)OH, trifluoromethyl, -C≡N, -C(=O)O(C-C)alkyl, -C(=O)NH, -SONH, -C(=NH)NH, and -NO, preferably containing one or two substituents selected from halogen, -OH, alkoxy, -NH, trifluoromethyl, -N(CH), and -C(=O)OH, more preferably halogen, alkoxy, and -OH. Examples of substituted alkyl include, but are not limited to, 2,2-difluoropropyl, 2-carboxycyclopentyl, and 3-chloropropyl.

[0030] A "lower alkyl" is an alkyl group, as defined above, having from 1 to 6 carbon atoms, preferably 1 to 4 carbon atoms, in its backbone structure, unless the number of carbon atoms is otherwise specified. A "lower alkenyl" and a "lower alkynyl" have a chain length of from 2 to 6 carbon atoms, preferably 2 to 4 carbon atoms.

[0031] "Alkenyl" includes unsaturated aliphatic groups similar in length and possible substitution to the alkyl groups described above, except that they contain at least one double bond. For example, the term "alkenyl" includes straight-chain alkenyl groups (e.g., ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl), branched-chain alkenyl groups, cycloalkenyl (e.g., alicyclic) groups (e.g., cyclopropenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl- or alkenyl-substituted cycloalkenyl groups, and cycloalkyl- or cycloalkenyl-substituted alkenyl groups. In certain cases, a straight-chain or branched-chain alkenyl group has six or fewer carbon atoms in its backbone (e.g., C2-C6 for straight chain, C3-C6 for branched chain). Likewise, cycloalkenyl groups have from 3-8 carbon atoms in their ring structure, and in some embodiments, cycloalkenyl groups have 5 or 6 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 3-6 carbon atoms.

[0032] "Alkynyl" includes unsaturated aliphatic groups analogous in length and possible substitution to the alkyl groups described above, except that they contain at least one triple bond. For example, "alkynyl" includes straight-chain alkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonyl, decynyl), branched-chain alkynyl groups, and cycloalkyl- or cycloalkenyl-substituted alkynyl groups. In certain embodiments, a straight-chain or branched-chain alkynyl group has six or fewer carbon atoms in its backbone (e.g., C2-C6 for straight chain, C3-C6 for branched chain). The term "C2-C6" includes alkynyl groups containing two to six carbon atoms. The term "C3-C6" includes alkynyl groups containing three to six carbon atoms.

[0033] The term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic radical in which each atom forming the ring (i.e., skeletal atoms) is a carbon atom. In some cases, cycloalkyl groups are saturated or partially unsaturated. In other cases, cycloalkyl groups are fused to an aromatic ring. Cycloalkyl groups include groups having 3 to 10 ring atoms. Examples of cycloalkyl groups include, but are not limited to, the following moieties:

[0034] [ka]

[0035] Monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Bicyclic cycloalkyls include, but are not limited to, tetrahydronaphthyl, indanyl, and tetrahydropentalene. Polycyclic cycloalkyls include adamantane and norbornane. The term cycloalkyl includes "unsaturated non-aromatic carbocyclyl" or "non-aromatic unsaturated carbocyclyl" groups, both of which refer to a non-aromatic carbocyclic ring containing at least one carbon-carbon double bond or one carbon-carbon triple bond, as defined herein.

[0036] The term "cycloalkylalkyl" refers to an alkyl group substituted with a cycloalkyl group. Examples of cycloalkylalkyl groups include cyclopropylalkyl and cyclohexylalkyl.

[0037] The term "heterocycloalkyl" refers to a non-aromatic heterocycle in which one or more ring-forming atoms are heteroatoms such as O, N, or S atoms. Heterocycloalkyl groups include monocyclic or polycyclic (e.g., 2, 3, or 4 fused rings) ring systems and spirocycles. Examples of heterocycloalkyl groups include morpholino, thiomorpholino, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, 2,3-dihydrobenzofuryl, 1,3-benzodioxole, benzo-1,4-dioxane, piperidinyl, pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, and imidazolidinyl. The definition of heterocycloalkyl can also include moieties having one or more aromatic rings fused to (i.e., sharing a bond with) a non-aromatic heterocycle, such as quinolyl, isoquinolyl, and benzo derivatives of heterocycles. Heterocycloalkyl groups having one or more fused aromatic rings are bonded through an aromatic or non-aromatic portion. The definition of heterocycloalkyl also includes moieties in which one or more ring-forming atoms can be substituted with one or two oxo or sulfido groups. In some cases, heterocycloalkyl groups have 1 to about 20 carbon atoms, and in further cases, about 3 to about 20 carbon atoms. In some cases, heterocycloalkyl groups contain 3 to about 20, 3 to about 14, 3 to 7, or 5 to 6 ring-forming atoms. In some cases, heterocycloalkyl groups contain 1 to about 4, 1 to about 3, or 1 to 2 heteroatoms. In some cases, heterocycloalkyl groups contain 0 to 3 double bonds. In some cases, heterocycloalkyl groups contain 0 to 2 triple bonds.

[0038] The term "heterocycloalkylalkyl" refers to an alkyl group substituted with a heterocycloalkyl. Examples of heterocycloalkylalkyl include morpholinoalkyl and piperazinylalkyl.

[0039] The term "aryl" refers to a monocyclic or polycyclic (e.g., 2, 3, or 4 fused rings) aromatic hydrocarbon, such as phenyl, naphthyl, anthracenyl, phenanthrenyl, etc. In some cases, aryl groups have from 6 to about 20 carbon atoms.

[0040] The term "arylalkyl" refers to an alkyl group substituted with an aryl group. Examples of arylalkyl groups include benzyl and phenylethyl.

[0041] The term "heteroaryl" refers to an aromatic heterocycle having at least one heteroatom ring member such as O, S, or N. Heteroaryl groups include monocyclic and polycyclic (e.g., 2, 3, or 4 fused ring) systems. Ring-forming N atoms in heteroaryl groups can be oxidized to form N-oxo moieties. Examples of heteroaryl groups include pyridyl, N-oxopyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furyl, quinolyl, isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl, pyrryl, oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl, isothiazolyl, benzothienyl, purinyl, carbazolyl, benzimidazolyl, and indolinyl. In some cases, heteroaryl groups have 1 to about 20 carbon atoms, and in further cases, from about 3 to about 20 carbon atoms. In some cases, heteroaryl groups contain 3 to about 14, 3 to 7, or 5 to 6 ring-forming atoms. In some cases, heteroaryl groups contain 1 to about 4, 1 to about 3, or 1 to 2 heteroatoms.

[0042] A "heteroarylalkyl" group refers to an alkyl group substituted with a heteroaryl group. An example of a heteroarylalkyl group is pyridylmethyl.

[0043] The term "halo" or "halogen" 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 "perhalogenated" refers to a moiety in which all hydrogens are replaced with halogens. The term "haloalkyl" refers to an alkyl moiety in which halogens replace hydrogens on one or more carbons of the hydrocarbon backbone. C1-C6 haloalkyl includes straight or branched alkyls having up to six main chain carbon atoms and halogens replacing hydrogens on one or more main chain carbons.

[0044] The terms "alkoxy" or "alkoxyl" include substituted and unsubstituted alkyl, alkenyl, and alkynyl groups covalently linked to an oxygen atom. C1-C6 alkoxy refers to a moiety having six or fewer carbon atoms in the hydrocarbon backbone. Examples of alkoxy groups (or alkoxyl radicals) include methoxy, ethoxy, isopropyloxy, propoxy, butoxy, and pentoxy groups. Preferred are (C1-C3) alkoxy, particularly ethoxy and methoxy. Examples of substituted alkoxy groups include halogenated alkoxy groups.

[0045] The term "hydroxy" or "hydroxyl" includes groups with an --OH or --O-- group.

[0046] The term "pharmaceutically acceptable salt" refers to a derivative of a compound in which the parent compound has been modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. Pharmaceutically acceptable salts include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or a mixture of the two; non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are usually preferred. Lists of suitable salts can be found 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, editors, Handbook of Pharmaceutical Salts: Properties, Selection and Use, 2nd Revised edition, Weinheim / Zurich: Wiley-VCH / VHCA (2011), each of which is incorporated herein by reference in its entirety.

[0047] Examples of suitable inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid, or hydrobromic acid, and 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, embonic 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, and examples of suitable organic bases include amines, for example tertiary amines such as trimethylamine, triethylamine, pyridine, N,N-dimethylaniline, quinoline, isoquinoline, α-picoline, β-picoline, γ-picoline, quinaldine or pyrimidine.

[0048] The term "antibody" encompasses the structure that constitutes the natural biological form of an antibody. In most mammals, including humans and mice, this form is a tetramer, consisting of two identical pairs of immunoglobulin chains, each pair having one light and one heavy chain, and each light chain containing immunoglobulin domain V. L and C L Each heavy chain contains immunoglobulin domain V H , Cγ1, Cγ2, and Cγ3. In each pair, the variable regions of the light and heavy chains (V L and V H ) together are responsible for binding to antigens, and the constant region (C L , Cγ1, Cγ2, and Cγ3, especially Cγ2 and Cγ3) are responsible for antibody effector functions. In some mammals, such as camels and llamas, full-length antibodies may consist of only two heavy chains, each containing immunoglobulin domain V. H, Cγ2, and Cγ3. As used herein, "immunoglobulin (Ig)" refers to a protein consisting of one or more polypeptides substantially encoded by immunoglobulin genes. Immunoglobulins include, but are not limited to, antibodies. Immunoglobulins include full-length antibodies, antibody fragments, and V H , Cγ1, Cγ2, Cγ3, V L and C L The immunoglobulin domains may have several structural forms, including individual immunoglobulin domains including:

[0049] Based on the amino acid sequence of the heavy chain constant domain, intact antibodies can be assigned to different "classes." There are five major classes (isotypes) of intact antibodies: IgA, IgD, IgE, IgG, and IgM, some of which can be further divided into "subclasses," e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The heavy chain constant domains corresponding to the various antibody classes are called alpha, delta, epsilon, gamma, and mu, respectively. The subunit structures and three-dimensional configurations of the various classes of immunoglobulins are well known to those skilled in the art.

[0050] The terms "antibody" or "antigen-binding fragment" refer to intact molecules and functional fragments thereof, respectively, such as Fab, scFv-Fc bivalent molecules, F(ab')2, and Fv, which are capable of specifically interacting with a desired target. In some cases, an antigen-binding fragment is (1) Fab, which is a fragment that contains a monovalent antigen-binding fragment of an antibody molecule and can be produced by digesting a whole antibody with the enzyme papain to yield an intact light chain and a portion of one heavy chain; (2) Fab', the fragment of an antibody molecule that can be obtained by treating whole antibody with pepsin, followed by reduction, to yield an intact light chain and a portion of the heavy chain; two Fab' fragments are obtained per antibody molecule; (3) (Fab')2, the fragment of an 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; (4) Fv, a genetically engineered fragment containing the light chain variable region and the heavy chain variable region expressed as two chains; (5) Single-chain antibodies ("SCAs"), which are genetically engineered molecules containing a light chain variable region and a heavy chain variable region linked by a suitable polypeptide linker as a genetically fused single-chain molecule; and (6) scFv-Fc, which is produced by fusing a single-chain Fv (scFv) with a hinge region and an Fc region derived from an immunoglobulin (Ig) such as IgG. Includes:

[0051] In one embodiment, the antibodies provided by the present invention are monoclonal antibodies. In one embodiment, the antigen-binding fragments provided by the present invention are single-chain Fvs (scFvs), diabodies, tandem scFvs, scFv-Fc bivalent molecules, Fabs, Fab's, Fvs, F(ab')2s, or antigen-binding scaffolds (e.g., affibodies, monobodies, anticalins, DARPins, Knottins).

[0052] The terms "bind," "binding," or grammatical equivalents refer to compositions that have a direct or indirect affinity for one another. "Specific binding" refers to when the binding between two molecules is selective. A specific example of specific binding occurs between an antibody and an antigen. Specific binding is usually measured by a dissociation constant (K D ) is about 1 × 10 -5 Less than M or approximately 1 × 10 -6 M or 1 x 10 -7M or less can be distinguished from nonspecific binding. Specific binding can be detected, for example, by ELISA, immunoprecipitation, co-precipitation, and two-hybrid assays, with or without chemical cross-linking. Appropriate controls can be used to distinguish "specific" from "nonspecific" binding. "Affinity" refers to the strength of the binding interaction between two molecules, such as an antigen and its antibody, and is defined as the strength of binding between a ligand and one specific binding site for antibodies and other molecules with two or more binding sites. Non-covalent binding between a ligand and an antibody or other molecule is usually not as strong as covalent binding, but "high affinity" can be defined as a binding strength of 10 as determined by inhibition ELISA. 4 M -1 Super, normal 10 5 ~10 11 M -1 The affinity constant (K a ) or equivalent techniques such as Scatchard plots, or by K a The reciprocal of K d / for a ligand that binds to an antibody or other molecule with an affinity comparable to this, as determined using the dissociation constant.

[0053] The term "selective" with respect to binding, inhibition, stimulation, or modulation means that a first activity binds, inhibits, stimulates, or modulates preferentially over a second activity (e.g., one receptor binds preferentially to another receptor; inhibits preferentially over other receptors; or inhibits a mutant over a wild-type receptor, or vice versa). In some cases, the binding is more than 2-fold, more than 5-fold, more than 10-fold, more than 50-fold, more than 100-fold, or more than 1000-fold selective for the desired molecular target or pathway over the undesired molecular target or pathway. In some cases, the compound binds to or affects a first molecular target or pathway at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 50-fold, or at least 100-fold more selectively than a second target or pathway under the same conditions. It will be understood that in preferred embodiments, binding to the D2-like family of dopamine receptors or members thereof will be more selective than the D1-like family of dopamine receptors or members thereof by any of the amounts described above. The in vitro or in vivo activity of a molecular target or pathway may be measured by any suitable reproducible means.

[0054] The term "modulation" refers to "stimulating" or "inhibiting" 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 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 compared to the activity of the target or pathway under the same conditions but without the composition. 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, or at least 100-fold compared to the activity of the target or pathway under the same conditions but without the composition. 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 any assay known in the art that is suitable for measuring that activity. A control sample (untreated with the composition) can be assigned a relative activity value of 100%.

[0055] In one embodiment, the antibody, antigen-binding fragment, or affinity tag has a K in the range of 0.1 nM to 10 mM, 0.1 nM to 1 mM, or 0.1 nM. D In one embodiment, the antibody, antigen-binding fragment, or affinity tag binds to its target with a K of 0.1-2 nM, 0.1-1 nM, 0.05-1 nM, 0.1-0.5 nM, or 0.1-0.2 nM. D In one embodiment, the antibody, antigen-binding fragment, or affinity tag binds to its target directly. In one embodiment, the antibody, antigen-binding fragment, or affinity tag binds to its target indirectly, for example, as a secondary antibody that binds to the antibody bound to the target.

[0056] The term "label" refers to a compound or composition that is directly or indirectly conjugated or fused to a reagent, such as a nucleic acid probe or antibody, and that renders the conjugated or fused reagent easier to detect. The label may be itself detectable (e.g., a radioisotope or fluorescent label) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition that is detectable.

[0057] The term "probe" refers to a synthetic or biologically produced nucleic acid containing a specific nucleotide sequence that hybridizes to a target nucleic acid sequence under stringent conditions. The terms "labeled probe," "nucleic acid probe operably linked to a detectable label," or "nucleic acid strand operably linked to a detectable label" refer to a probe prepared with a marker moiety or "detectable label" for detection. The marker moiety may be attached to the 5' end, 3' end, internally, or a combination thereof. That is, a single probe may be attached to multiple marker moieties. Preferred moieties include distinguishing labels such as fluorophores. A labeled probe may also contain multiple different nucleic acid sequences labeled with one or more marker moieties. Each marker moiety may be the same or different. It may be advantageous to label different probes (e.g., nucleic acid sequences) with different marker moieties. This can be achieved by providing a single distinguishing moiety on each probe. For example, probe A may be attached to moiety X and probe B to moiety Y. Alternatively, probe A may be attached to moieties X and Y, and probe B to moieties Z and W. Alternatively, probe A is conjugated to moieties X and Y, and probe B is conjugated to moieties Y and Z. Both probes "A" and "B" above will be distinguishably and uniquely labeled.

[0058] A "tissue sample" refers to a collection of similar cells, preferably containing nucleated cells with chromosomal material, taken from the 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 source of the tissue sample can be fresh, frozen, and / or preserved organ or tissue samples or solid tissue obtained from biopsy or aspiration; blood or blood components; bodily fluids such as cerebrospinal fluid, amniotic fluid, peritoneal fluid, or interstitial fluid; or cells from a subject's pregnancy or development. A tissue sample can be primary or cultured cells or cell lines. A tissue sample can contain compounds that are not naturally associated with the tissue in its natural state, such as preservatives, anticoagulants, buffers, fixatives, nutrients, or antibiotics. A "section" of a tissue sample refers to a single portion or piece of the tissue sample, e.g., a thin slice of tissue or cells cut from the tissue sample. Multiple sections of a tissue sample can be taken and subjected to analysis. "Cell line" refers to a permanently established cell culture that will grow if given adequate fresh medium and space.

[0059] Detection Method

[0060] In various embodiments, the present invention provides methods for detecting or measuring a target receptor (e.g., a dopamine receptor or GPCR) in a biological sample. The target is detected by contacting the sample with a target detection reagent, such as 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 target 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 together with the labeling reagent, under conditions in which a complex is formed between the detection reagent (and labeling reagent) and the target. After complex formation, the sample is optionally washed one or more times to remove unbound detection reagent (and labeling reagent). If the sample is further contacted with a reagent that specifically binds to the detection reagent bound to the target, the sample may 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.

[0061] The methods described herein detect multiple targets in a sample. Using the methods described, the multiple targets are identified by contacting the biological sample with an additional detection reagent and then an additional labeling reagent specific for the additional detection reagent.

[0062] A detection moiety, or detectable label, is a substance that facilitates target identification and / or quantification. Detection moieties can be directly observed or measured, or indirectly observed or measured. Detection moieties include, but are not limited to, radiolabels that can be measured with a radiation measuring device; dyes, pigments, or other chromogens that can be visually observed or measured with a spectrophotometer; spin labels that can be measured with a spin label analyzer; and fluorescent moieties, where excitation of an appropriate molecular adduct generates an output signal that can be visualized by excitation with light absorbed by the dye or measured with a standard fluorometer or imaging system. Detection moieties can be luminescent substances such as fluorescent substances or fluorophores; bioluminescent substances; chemiluminescent substances that generate an output signal by chemical modification of the signal compound; metal-containing substances; or enzymes that result in enzyme-dependent secondary signal generation, such as the formation of a colored product from a colorless substrate. Detection moieties can also take the form of chemical or biochemical particles or inert particles, including inorganic crystals such as colloidal gold, microspheres, quantum dots, nanocrystals, or fluorophores. The term detection moiety or detectable label can also refer to a "tag" or hapten that can selectively bind to a labeled molecule and generate a detectable signal using a subsequently added labeled molecule. For example, biotin, iminobiotin, or desthiobiotin can be used as a tag, and then horseradish peroxidase (HRP) can be conjugated to the tag using an avidin or streptavidin conjugate, followed by detection of the presence of HRP using a chromogenic substrate (e.g., tetramethylbenzidine) or a fluorogenic substrate such as Amplex Red or Amplex Gold (Molecular Probes). Similarly, the tag can be a hapten or antigen (e.g., digoxigenin), and can be conjugated to the tag using an antibody labeled with an enzyme, a fluorescent substance, or a radioactive substance. Numerous labels are known to those skilled in the art, including, but not limited to, particles, fluorescent dyes, haptens, enzymes, and their chromogenic, fluorogenic, and chemiluminescent substrates.

[0063] Fluorophores are chemical moieties that exhibit an absorption maximum above 280 nm and retain their spectral properties when covalently attached to 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-1,3-diazole (NBD), carbocyanine, carbostyryl, salicylate, anthranilate, azulene, pyridine, quinoline, borapolyazaindacene, xanthene, oxazine or benzoxazine, carbazine, phenalenone, coumarin, benzofuran, and benzphenalenone, as well as their derivatives. Oxazines include resorufin, aminooxazinones, diaminooxazines, and their benzo-substituted analogs.

[0064] When the fluorophore is a xanthene, it can be fluorescein, rhodol, or rhodamine. Fluoresceins include benzofluorescein or dibenzofluorescein, seminaphthofluorescein, or naphthofluorescein. Similarly, rhodols include seminaphthorhodafluor. Alternatively, the fluorophore is a xanthene bound via a single covalent bond at the 9-position of the xanthene. Preferred xanthenes include derivatives of 3H-xanthen-6-ol-3-one, derivatives of 6-amino-3H-xanthen-3-one, or derivatives of 6-amino-3H-xanthen-3-imine. Fluorophores include xanthenes (rhodol, rhodamine, fluorescein, and their derivatives), coumarins, cyanines, pyrenes, oxazines, and borapolyazaindacenes. Furthermore, the fluorophore can be a sulfonated xanthene, a fluorinated xanthene, a sulfonated coumarin, a fluorinated coumarin, or a sulfonated cyanine. The choice of fluorophore for a labeling reagent determines the absorption and fluorescence emission properties of the labeling reagent. The physical properties of the fluorophore label include spectral properties (absorption, emission, and Stokes shift), fluorescence intensity, lifetime, polarization, and photobleaching rate, all of which can be used to distinguish one fluorophore from another.

[0065] Fluorophores typically contain one or more aromatic or heteroaromatic rings that are optionally substituted with one or more of a variety of substituents, including halogen, nitro, cyano, alkyl, perfluoroalkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, arylalkyl, acyl, aryl or heteroaryl ring systems, benzo, or other substituents commonly found on fluorophores and known in the art.

[0066] Preferably, the detection moiety is a fluorescent dye. Examples of fluorescent dyes include fluorescein, rhodamine, Texas Red, Cy2, Cy3, Cy5, Cy0, Cy0.5, Cy1, Cy1.5, Cy3.5, Cy7, VECTOR Red, ELF™ (Enzyme-Labeled Fluorescence), FluorX, Calcein, Calcein-AM, CRYPTOFLUOR™'S, Orange (42 kDa), Tangerine (35 kDa), Gold (31 kDa), Red (42 kDa), Crimson (40 kDa), BHMP, BHDMAP, Br-Oregon, Lucifer Yellow, the Alexa dye family, N-(6-(7-nitrobenzo-2-oxa-1,3-diazol-4-yl)amino)caproyl (NBD), BODIPY™, borondipyrromethene difluoride, and Oregon Green, MITOTRACKER™ Red, 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 function as chromophores, and therefore are also preferred chromophores.

[0067] In addition to fluorophores, enzymes are also used as detectable moieties. Enzymes are desirable detectable moieties because they can increase assay sensitivity by amplifying the detectable signal. While the enzyme itself does not produce a detectable response, upon contact with an appropriate substrate, it cleaves the substrate, generating a fluorescent, colorimetric, or luminescent signal from the converted substrate. Enzymes amplify the detectable signal because a single enzyme for a given labeling reagent converts multiple substrates into detectable signals. This is advantageous when the amount of target present in a sample is low or when no fluorophore is available that generates a signal as strong or stronger than the enzyme. However, fluorophores are preferred because they do not require additional assay steps, thereby reducing the overall time required to complete the assay. The enzyme substrate is selected to produce a desired measurable product, such as colorimetric, fluorescent, or chemiluminescent. Such substrates are widely used in the art.

[0068] A preferred chromogenic or fluorogenic substrate-enzyme combination is one that uses an oxidoreductase, such as horseradish peroxidase, with substrates such as 3,3'-diaminobenzidine (DAB) and 3-amino-9-ethylcarbazole (AEC), which produce characteristic colors (brown and red, respectively). Other chromogenic oxidoreductase substrates that produce detectable products include, but are not limited to, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), o-phenylenediamine (OPD), 3,3',5,5'-tetramethylbenzidine (TMB), o-dianisidine, 5-aminosalicylic acid, and 4-chloro-1-naphthol. Fluorogenic substrates include, but are not limited to, homovanillic acid or 4-hydroxy-3-methoxyphenylacetic acid, reduced phenoxazines and benzothiazines, including Amplexe Red reagent and its variants, and reduced dihydroxyxanthenes, including dihydrofluorescein and dihydrorhodamine 123. Tyramide peroxidase substrates are a unique class of peroxidase substrates in that they may be intrinsically detectable before the action of the enzyme, but are "locked in place" by the action of peroxidase in a process called tyramide signal amplification (TSA). These substrates are widely used to label targets in cell, tissue, or array samples for subsequent detection by microscopy, flow cytometry, optical scanning, and fluorometry.

[0069] Additional chromogenic (and in some cases fluorogenic) substrate and enzyme combinations utilize phosphatase enzymes, such as acid phosphatase, alkaline phosphatase, or recombinant forms of such phosphatases, in combination with chromogenic 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 fluorogenic substrates such as 4-methylumbelliferyl phosphate, 6,8-difluoro-7-hydroxy-4-methylcoumarinyl phosphate (DiFMUP), fluorescein diphosphate, 3-O-methylfluorescein phosphate, resorufin phosphate, 9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl) phosphate (DDAO phosphate), or ELF97, ELF39, or related phosphates.

[0070] Glycosidases, particularly β-galactosidase, β-glucuronidase, and β-glucosidase, are further suitable enzymes. Suitable chromogenic substrates include, but are not limited to, 5-bromo4-chloro-3-indolyl β-D-galactopyranoside (X-gal) and similar indolyl galactosides, glucosides, and glucuronides, o-nitrophenyl β-D-galactopyranoside (ONPG) and p-nitrophenyl β-D-galactopyranoside. Preferred fluorogenic substrates include resorufin β-D-galactopyranoside, fluorescein diglucuronide (FDG), fluorescein diglucuronide and its structural variants, 4-methylumbelliferyl β-D-galactopyranoside, carboxyumbelliferyl β-D-galactopyranoside, and fluorinated coumarin β-D-galactopyranoside. Further enzymes include hydrolases such as cholinesterases and peptidases, oxidative enzymes such as glucose oxidase and cytochrome oxidase, and reductases for which suitable substrates are known.

[0071] Chemiluminescent enzymes and suitable substrates are preferred for some assays. These include, but are not limited to, natural and recombinant luciferases and aequorins. Additionally, chemiluminescent substrates for phosphatases, glycosidases, and oxidases, such as those containing stable dioxetanes, luminol, isoluminol, and acridinium esters, are useful. For example, the enzyme is luciferase or aequorin. The substrates can be luciferin, ATP, Ca, or ATP. ++ and coelenterazine.

[0072] In addition to enzymes, haptens such as biotin are also useful detectable moieties. Biotin is useful because it exists in enzyme systems that can further amplify the detectable signal and can serve as a tag for affinity chromatography for isolation. Detection is performed using an enzyme conjugate with affinity for biotin, such as avidin-HRP. A peroxidase substrate is then added to generate a detectable signal. Haptens also include hormones, natural and synthetic drugs, pollutants, allergens, affector molecules, growth factors, chemokines, cytokines, lymphokines, amino acids, peptides, chemical intermediates, or nucleotides.

[0073] In some cases, the detectable moiety is a fluorescent protein. Exemplary fluorescent proteins include green fluorescent protein (GFP), phycobiliproteins and their derivatives, luciferase, or aequorin. Fluorescent proteins, particularly phycobiliproteins, are particularly useful for creating tandem dye-labeled labeling reagents. These tandem dyes contain a fluorescent protein and a fluorophore to achieve a larger Stokes shift, where the emission spectrum is shifted further away from the absorption spectrum of the fluorescent protein. This is particularly advantageous for detecting low-abundance targets in a sample when the emitted fluorescence is maximally optimized, i.e., when the fluorescent protein reabsorbs little or no emitted light. The fluorescent protein and fluorophore function as an energy transfer pair, where the fluorescent protein emits at a wavelength where the fluorophore absorbs, and the fluorphore then emits at a wavelength further away from the fluorescent protein than would be possible with the fluorescent protein alone. A particularly useful combination is a phycobiliprotein with a sulforhodamine fluorophore or a sulfonated cyanine fluorophore; or a sulfonated xanthene derivative. Alternatively, the fluorophore is the energy donor and the fluorescent protein is the energy acceptor.

[0074] The method for visualizing the detection moiety depends on the label

[0075] In some cases, the sample is exposed to a wavelength of light selected to produce a detectable optical response, and the response is observed using a means for detecting it. Useful devices for illuminating fluorescent compounds include portable 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 microfluorometers. The level or position of the signal compared to the baseline response or expected response indicates the presence or absence and degree of a given characteristic or desired target in the sample.

[0076] The optical response is detected visually or by the following devices: CCD camera, video camera, photographic film, laser scanning device, fluorometer, photodiode, quantum counter, epifluorescence microscope, scanning microscope, flow cytometer, fluorescence microplate reader, or by means of signal amplification such as a photomultiplier tube. When the sample is examined using a flow cytometer, the examination optionally includes aliquoting portions according to their fluorescence response.

[0077] When an indirectly detectable label is used, illumination usually involves adding a reagent to generate a detectable signal, such as a chromogenic enzyme substrate. Radioisotopes are also considered indirectly detectable, in which case no additional reagents are required; rather, the signal is recorded and measured by exposing the radioisotope to X-ray film or other mechanism. This is also true for some chemiluminescent signals, which are observed after exposure to film.

[0078] I. ONC201 (Compound (1)), its salts and their synthesis

[0079] ONC201 (compound (1))

[0080] [ka]

[0081] The present invention provides ONC201 and its analogs, pharmaceutically acceptable salts, and syntheses thereof. In vitro models, animal models, and human clinical trials have shown that ONC201 has broad-spectrum anticancer activity and possesses low toxicity with few, if any, adverse effects, low genotoxicity, and high bioavailability, including oral bioavailability. These characteristics make ONC201 and various analogs suitable for a variety of applications. ONC201 can be prepared by the synthesis shown in Scheme 1 below.

[0082] [ka]

[0083] The synthesis of ONC201 dihydrochloride begins with the commercially available intermediate N-benzyl-3-carbomethoxy-4-piperidone hydrochloride, compound (3). In one embodiment, the synthesis involves neutralizing compound (3) with a base (Step 1) to form the free base of compound (4). In one embodiment, compound (3) is neutralized with an inorganic base to form compound (4). In one embodiment, compound (3) is neutralized with an organic base to form compound (4). In one embodiment, compound (3) is neutralized in the presence of an alcohol, such as n-butanol. In one embodiment, compound (3) is neutralized in the presence of at least one organic solvent, such as 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, such as NaHCO and n-butanol. In one embodiment, compound (3) is neutralized in the presence of n-butanol and triethylamine (EtN).

[0084] In one embodiment, the synthesis involves reacting compound 4 with compound 5 (Step 2) to produce intermediate compound 1. In one embodiment, the Step 2 reaction involves heating compound 4 with compound 5. In one embodiment, the Step 2 reaction involves heating compound 4 and compound 5 to reflux in the presence of a solvent. In one embodiment, the Step 2 reaction involves the use of a Dean-Stark trap to remove water and / or methanol (MeOH) formed during the reaction.

[0085] In one embodiment, ONC201 dihydrochloride is synthesized (Step 3). In one embodiment, this reaction (Step 3) involves treating ONC201 with HCl in dioxane. In one embodiment, Step 3 involves treating ONC20 with 4N HCl in dioxane. In one embodiment, the synthesis optionally involves recrystallization of the ONC201 dihydrochloride. In a preferred embodiment, ONC201 dihydrochloride is synthesized as shown in Scheme 2 below.

[0086] [ka]

[0087] II. TNF-Related Apoptosis-Inducing Ligand ("TRAIL")

[0088] TRAIL protein can be assayed in the sample collected from the subject to detect the TRAIL expression induced by the compounds described herein and their salts.TRAIL can be assayed in the sample using immunoassay methods, including enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunofiltration assay (ELIFA), flow cytometry, immunoblot, immunoprecipitation, immunohistochemistry, immunocytochemistry, luminescence immunoassay (LIA), fluorescence immunoassay (FIA) and radioimmunoassay.The assay can be used to obtain qualitative and / or quantitative results. Specific details regarding methods suitable for both qualitative and quantitative sample assays can be found in E. Harlow & D. Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1988; F. Breitling & S. Diibel, 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; BKC Lo, Antibody Engineering: Methods and Protocols, Methods in Molecular Biology, Humana Press, 2003; FMAusubel 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,MGIt is described in standard references, including Givan, A.L., 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, 3rd Edition, Elsevier Science, 2005, and J. Sambrook and D.W. Russell, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 3rd ed., 2001.

[0089] Protocols for assaying and analyzing TRAIL in samples to detect the effects of pharmaceutical compositions are described in U.S. Patent No. 8,673,923 to Wafik S. El-deiry et al., which is incorporated herein by reference in its entirety.

[0090] In one embodiment, a TRAIL assay is used to monitor a subject. For example, samples are taken from a subject before treatment with a pharmaceutical composition, and at one or more time points during and / or after treatment to assess the effectiveness of the treatment. In another example, samples are taken from a subject at various time points to assess the course or progression of disease or healing. In one embodiment, death receptors in circulating tumor cells are assayed to determine whether the treatments described herein increase the amount or type of death receptors.

[0091] Cancers that can be treated using the methods and compositions described herein are characterized by abnormal cell proliferation, including pre-neoplastic hyperproliferation, in situ cancer, neoplasia, and metastasis. The methods and compositions described herein can be used to prevent and ameliorate signs and / or symptoms of cancer. "Treating" a subject's cancer includes preventing, inhibiting, or ameliorating the subject's cancer, such as slowing the progression of cancer, or alleviating or ameliorating signs or symptoms of cancer. Examples of cancers that can be treated using the methods and compositions described herein include breast cancer, CNS cancer, colon cancer, ovarian cancer, prostate cancer, leukemia, lung cancer, and lymphoma.

[0092] III. Compounds of structural formula (10) and salts thereof

[0093] In one aspect, the present invention provides compounds and salts of structural formula 10 and methods for their preparation. Those skilled in the art will appreciate that the general principles and concepts described herein in conjunction with ONC201 (compound 1) and its salts, including principles and concepts related to methods and pharmaceutical compositions, apply equally to compounds of structural formula 10 and their salts.

[0094] In one embodiment, the present invention provides a compound represented by the following structural formula (10):

[0095] [ka]

[0096] wherein R1 and R2 are independently selected from H, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, arylalkyl, heteroarylalkyl, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, and acyl radicals. In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2-CH3-Ph) (i.e., ONC201). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2,4-diF-Ph) (i.e., ONC206). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(4-CF3-Ph) (i.e., ONC212). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(3,4-diF-Ph) (i.e., ONC213). In one embodiment, R1 is CH2(3,4-di-Cl-Ph) and R2 is CH2-(4-CF3-Ph) (i.e., ONC234). In one embodiment, R1 is CH2-3-thienyl and R2 is CH2-(4-CF3-Ph) (i.e., ONC236).

[0097] In one embodiment, R1 and R2 are H, 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 Alkylpyridinyl, C 1~4 Alkylisoxazolidinyl, C 1~4 Alkylmorpholinyl, C 1~4 Alkylthiazolyl and C 1~4 alkylpyrazinyl, independently selected from the group consisting of 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 Alkylpyridinyl, C 1~4 Alkylisoxazolidinyl, C 1~4Alkylmorpholinyl, C 1~4 Alkylthiazolyl and C 1~4 Alkylpyrazinyl is optionally C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, R and / or R are substituted or unsubstituted arylalkyl or heteroarylalkyl. In one embodiment, heteroarylalkyl is C 1~4 Alkylpyrrolyl, C 1~4 Alkylfuryl, C 1~4 Alkylpyridyl, C 1~4 Alkyl-1,2,4-thiadiazolyl, C 1~4 Alkylpyrimidyl, C 1~4 Alkylthienyl, C 1~4 Alkylisothiazolyl, C 1~4 Alkyl imidazolyl, C 1~4 Alkyltetrazolyl, C 1~4 Alkylpyrazinyl, C 1~4 Alkylpyrimidyl, C 1~4 Alkylquinolyl, C 1~4 Alkylisoquinolyl, C 1~4 Alkylthiophenyl, C 1~4 Alkylbenzothienyl, C 1~4 Alkylisobenzofuryl, C 1~4 Alkylpyrazolyl, C 1~4 Alkyl indolyl, C 1~4 Alkylpurinyl, C 1~4 Alkylcarbazolyl, C 1~4 Alkylbenzimidazolyl and C 1~4 alkylisoxazolyl.

[0098] In one embodiment, R1 and / or R2 optionally have one or more of the following substituents on the benzyl ring: 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, -OCpH2p+1 , -OC p X 2p+1 , OR m , S.R. m , N.R. m R n , N.R. m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m benzyl substituted with R m and R n are independently selected from H or C1-C4 alkyl; 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.

[0099] In one embodiment, R1 is 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-pyridinyl, CH2-4-methyl-2-thiazolyl, CH2-2-pyrazinyl, CH2CH2Ph, CH2CH 2(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, R2 is 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-CH 3)-Ph), CH2(2-CH3,4-F-Ph), CH2-((4-OCH3)-Ph), CH2-(3-pyridinyl), 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.

[0100] In one embodiment, R is H. In one embodiment, R is a substituted or unsubstituted arylalkyl, such as a benzyl (CHPh) or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 It is substituted with alkyl or halo.

[0101] In one embodiment, R2 is substituted or unsubstituted arylalkyl, such as benzyl or phenylethyl. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, the arylalkyl is substituted with one or more substituents selected from halo, CH3, CF3, or OCH3. In one embodiment, R2 is a substituted or unsubstituted heterocycloalkylalkyl, such as piperazinylalkyl or morpholinoalkyl. In one embodiment, R2 is a substituted or unsubstituted heteroarylalkyl, such as pyridylmethyl or isoxazolidinylmethyl. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with at least one substituent selected from halo, CH3, CF3, or OCH3.

[0102] In one embodiment, compound (10) has a structure represented by formula (80):

[0103] [ka]

[0104] In the formula, R a1 , R a2 , R a3 , R a4 , R a5 , R b1 , R b2 , R b3 , R b4 and R b5is 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 , S.R. m , N.R. m R n , N.R. m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m are each independently selected from the group consisting of: m and R n are independently selected from H or C1-C4 alkyl; p is an integer from 2 to 20, and X is a halogen.

[0105] In one embodiment, compound (10) has a structure represented by structural formula (90):

[0106] [ka]

[0107] where R2 is as defined above and R b1 , R b2 , R b3 , R b4 and R b5 is 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 , S.R. m , N.R. m Rn , N.R. m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m R m and R n is H or C 1~4 alkyl; p is an integer from 2 to 20, and X is a halogen.

[0108] In one embodiment, compound (10) has a structure represented by structural formula (40):

[0109] [ka]

[0110] where R1 is as defined above and R a1 , R a2 , R a3 , R a4 and R a5 is 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 , S.R. m , N.R. m R n , N.R. m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m R m and R n is H or C 1~4alkyl; p is an integer from 2 to 20; and X is halogen. In one embodiment, R1 is H. In one embodiment, R1 is substituted or unsubstituted arylalkyl, such as benzyl or phenylethyl. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 It is substituted with alkyl or halo. In one embodiment, benzyl is substituted with one or more halo. In one embodiment, benzyl is substituted with one or more substituents selected from halo, CH3, CF3 and OCH3. In one embodiment, benzyl is substituted with one halo, for example, F at the ortho or para position. In one embodiment, benzyl is substituted with two halogens, for example, F at both meta positions.

[0111] In one embodiment, compound 40 has a structure represented by formula 45:

[0112] [ka]

[0113] In the formula, R a1 , R a2 , R a3 , R a4 and R a5 is as defined above. In one embodiment, the benzyl is substituted with one or more halogens. In one embodiment, the benzyl is substituted with one or more substituents selected from halo, CH, CF, and OCH. In one embodiment, R a1 or R a5 is halo, e.g., F. In one embodiment, R a2 and R a3 are both halo, e.g. F.

[0114] In one embodiment, compound (10) has a structure represented by formula (50):

[0115] [ka]

[0116] where R1 is as defined above and R b is 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 , S.R. m , N.R. m R n , N.R. m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m R m and R n is H or C 1~4 alkyl; p is an integer from 2 to 20; X is halogen; and R a1 , R a2 , R a4 and R a5 is 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 , S.R. m , N.R. m R n , N.R. m C(O)R n , SOR m , SO2R m , C(O)R mand C(O)OR m are each independently selected from the group consisting of: m and R n is 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, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, R b is selected from halo, CH, CF, and OCH. In one embodiment, R a1 , R a2 , R a4 and R a5 One or more of R are selected from halo, CH, CF, and OCH. a1 , R a2 , R a4 and R a5 is H and R b is selected from halo, CH, CF, and OCH. In one embodiment, R b is a halogen, e.g., 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 is CF. In one embodiment, R b is OCH. In one embodiment, R b and R a1 is Cl.

[0117] In one embodiment, compound (50) has a structure represented by structural formula (55):

[0118] [ka]

[0119] In the formula, R a1 , R a2 , R a4 , R a5 and R b is as defined above. In one embodiment, R b is selected from halo, CH, CF, and OCH. In one embodiment, R a1 , R a2 , R a4 and R a5 One or more of R are selected from halo, CH, CF, and OCH. a1 , R a2 , R a4 and R a5 is H and R b is selected from halo, CH, CF, and OCH. In one embodiment, R b is a halo, e.g., 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 is CF. In one embodiment, R b is OCH. In one embodiment, R b and R a1 is Cl.

[0120] In one embodiment, compound (10) has a structure represented by formula (60):

[0121] [ka]

[0122] In one embodiment, R1 is H. In one embodiment, R1 is substituted or unsubstituted arylalkyl, such as benzyl or phenylethyl. 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-benzyl-piperazine), CH2-(3-isoxazolidinyl), CH2-2-pyridinyl, CH2-3-pyridinyl, and CH2CH2-(4-morpholinyl). In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, benzyl is substituted with one or more halogens. In one embodiment, benzyl is substituted with one or more substituents selected from halo (e.g., F), CH, CF, and OCH. In one embodiment, benzyl is substituted at the para position with a halo, CH, CF, or OCH substituent. In one embodiment, R is fluorophenyloxobutyl or hydroxyphenylethyl.

[0123] Scheme 3 below illustrates the synthesis of the compound of formula (10).

[0124] [ka]

[0125] Methods: a. NaH, dimethyl carbonate, toluene, 80°C, 4 hours; b. Convert to free base with 1N NaOH / CHCl, then heat in dioxane at 70°C; c. 1-butanol / reflux (Dean-Stark trap) PPTS; d. dioxane at 70°C; e. HCl in dioxane at -25°C to room temperature to make HCl salt; f. NaCO, DIEA at 80°C; g. Free base with NaOH / CHCl, then reflux MeOH, 3.5 hours

[0126] Starting from a substituted piperidone, compounds of structural formula (10) (i.e., imipridone) can be synthesized, which can be converted by reaction with a substituted aminoimidazoline to give the core compound (10). Two routes are available, one of which involves the R1 substituent present in the piperidone (e.g., 68). In this route, 68 is acylated with dimethyl carbonate using sodium hydride in toluene at 80 °C to form the piperidone ester (69). Commercially available methylthioimidazoline HI salt (63) is reacted with an amine in dioxane at 70 °C to give the R2-substituted aminoimidazoline (64) as its HI salt. Direct reaction of 64 with piperidone ester (69) in 1-butanol at reflux for 3–6 h with water removal using a Dean-Stark trap affords the tricyclic compound (10). In a variation of this scheme, N-BOC-protected piperidone 61 can be converted to BOC-protected compound 65 in the same manner, which can be treated with HCl in dioxane to remove the BOC group and then converted to the free base 66 by using 1N NaOH and extraction with methylene chloride. 66 can then be treated with halide 67 or epoxide 70 to give the desired compound 10.

[0127] The crude product can be purified by column chromatography eluting with methylene chloride:methanol or HPLC using acetonitrile:TFA:HO to obtain the final product as the free base or TFA salt. Treatment of the free base with HCl in dioxane or lyophilization of the TFA salt affords product (10) as the HCl or TFA salt. Alternatively, the free base may be treated with another inorganic or organic acid to form other salts selected from those generally known to be pharmaceutically acceptable. Salts of compound (10) are typically solids, and several examples have been crystallized from ethanol or other solvents to yield high-quality crystals. In the case of compound (1), the tricyclic structure has been conclusively confirmed by X-ray crystallography and NMR.

[0128] The compounds described herein, with or without an aminoalkyl linker (e.g., compound (33)), can be used to identify molecules (e.g., proteins) that interact with them in a particular cellular context. Expression of these binding targets can be used to predict response to imipridone or its analogs (i.e., serve as biomarkers). 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. Furthermore, these molecules may have improved drug properties or enable additional uses by altering drug properties, including safety, efficacy, pharmacokinetics, biodistribution, or metabolism.

[0129] [Table 1]

[0130] IV. Evaluating the sensitivity and efficacy of treatment regimens

[0131] Measurement of dopamine receptor or other G protein-coupled receptor (GPCR) expression, genetic mutation, or gene copy number may be used to predict response or susceptibility to the treatment methods described herein and to identify subjects likely to respond to the treatment methods described herein, such as treatment with a compound of structural formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one aspect, the present invention provides a method for determining whether a subject having a pathological condition is likely to respond to a treatment regimen described herein. In one embodiment, the method includes: (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 with that of a predetermined standard; and (iv) determining whether the subject is likely to respond to the treatment regimen based on the level measured in the sample relative to that of the predetermined standard. In one embodiment, measuring the expression level of a dopamine receptor or GPCR in a sample comprises (i) contacting the sample with an antibody or antigen-binding fragment that specifically binds to the receptor to form a complex between 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 cancer. In one embodiment, the cancer is a neuro-oncological 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, and 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, and 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 a bacterial infection 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 Staphylococcus infection. In one embodiment, the Staphylococcus infection is a S. aureus infection (e.g., a methicillin-resistant S. aureus (MRSA) infection). In one embodiment, the treatment regimen comprises administering an effective amount of a therapeutic agent, such as a compound of structural formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is a member of the D2-like family of dopamine receptors. 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 administering an effective amount of a therapeutic agent, such as a compound of structural formula (10) or a pharmaceutically acceptable salt thereof, and an increase in DRD5 expression level measured in the sample above a predetermined standard indicates that the subject is likely or unlikely to respond to the treatment regimen.

[0132] In another aspect, the present invention provides a method for evaluating the effectiveness of a treatment regimen described herein, for monitoring, or for predicting the prognosis of a subject with a condition. In one embodiment, the method includes: (i) collecting 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 with that of a predetermined standard; and (iv) determining the prognosis or whether the subject will respond to the treatment regimen based on the level measured in the sample relative to that of the predetermined standard. In one embodiment, measuring the expression level of a dopamine receptor or GPCR in the sample includes: (i) contacting the sample with an antibody or antigen-binding fragment that specifically binds to the receptor to form a complex between the antibody or antigen-binding fragment and the receptor; and (ii) measuring the amount of the complex. In one embodiment, the method includes (i) obtaining a biological sample from a subject; (ii) measuring gene copy numbers or mutations of at least one dopamine receptor in the sample; (iii) comparing the measured copy numbers or mutations found in the sample with those of a predetermined standard; and (iv) determining whether the subject will respond to a treatment regimen based on the measured copy numbers or mutations found in the sample relative to those of the predetermined standard. In one embodiment, the subject has or is at risk of having cancer. In one embodiment, the cancer is a neuro-oncological 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, and 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, and 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 a bacterial infection 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 Staphylococcus infection. In one embodiment, the Staphylococcus infection is a S. aureus infection (e.g., a methicillin-resistant S. aureus (MRSA) infection). In one embodiment, the treatment regimen comprises administering an effective amount of a therapeutic agent, such as a compound of structural 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 family of dopamine receptors. In one embodiment, the dopamine receptor is from the D1-like family of dopamine receptors. 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.

[0133] In one embodiment, the dopamine receptor is DRD5, and the treatment regimen comprises administering an effective amount of a compound of structural formula (10) or a pharmaceutically acceptable salt thereof, and an increase in expression level measured in the sample above a predetermined standard indicates that the treatment regimen is efficacious or ineffective. In one embodiment, the dopamine receptor is DRD5, and the treatment regimen comprises administering an effective amount of a therapeutic agent, e.g., a compound of structural formula (10) or a pharmaceutically acceptable salt thereof, and a mutation in the DRD5 gene measured in the sample indicates that the treatment regimen is efficacious or ineffective. In one embodiment, the dopamine receptor is DRD5, and the treatment regimen comprises administering an effective amount of a therapeutic agent, e.g., a compound of structural formula (10) or a pharmaceutically acceptable salt thereof, and a misense mutation Q366R in the DRD5 gene measured in the sample indicates that the treatment regimen is efficacious or ineffective.

[0134] In another aspect, the present invention provides a method for determining whether a subject with a pathological condition is likely to respond to the treatment regimen described herein. In one embodiment, the method includes: (i) collecting a biological sample from the subject; (ii) measuring the gene copy number or mutations of at least one dopamine receptor in the sample; (iii) comparing the measured copy number or found mutations in the sample with those of a predetermined standard; and (iv) determining whether the subject is likely to respond to the treatment regimen based on the measured copy number or found mutations relative to those of the predetermined standard. In one embodiment, the subject has or is at risk of having cancer. In one embodiment, the cancer is a neuro-oncological 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, and 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, and 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 a bacterial infection 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 Staphylococcus infection. In one embodiment, the Staphylococcus infection is a S. aureus infection (eg, a methicillin-resistant S. aureus (MRSA) infection).In one embodiment, the treatment regimen comprises administering an effective amount of a therapeutic agent, such as a compound of structural formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. In one embodiment, the dopamine receptor is of the D2-like family of dopamine receptors. 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, and the treatment regimen comprises administering an effective amount of a therapeutic agent, such as a compound of structural formula (10), or a pharmaceutically acceptable salt thereof, and a mutation in the DRD5 gene measured in the sample indicates that the subject is likely or unlikely to respond to the treatment regimen. 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 structural formula (10) or a pharmaceutically acceptable salt thereof, and a misense mutation Q366R in the DRD5 gene measured in the sample indicates that the subject is likely or unlikely to respond to the treatment regimen.

[0135] Furthermore, measurements of the expression, post-translational modifications, activity levels, or mutations of eIF2-α, ATF4, CHOP, DR5, or truncated or intact cytokeratin 18 can be used to predict response or susceptibility to the therapeutic methods described herein and to identify subjects likely to respond to the therapeutic methods described herein, such as treatment with a compound of structural formula (10), a pharmaceutically acceptable salt thereof, or an analog thereof. Furthermore, measurements of the expression, post-translational modifications, activity levels, or mutations of eIF2-α, ATF4, CHOP, DR5, or truncated or intact cytokeratin 18 can be used to evaluate the efficacy of or monitor the therapeutic methods described herein. Furthermore, measurements of the expression, post-translational modifications, activity levels, or mutations of eIF2-α, ATF4, CHOP, DR5, or truncated or intact cytokeratin 18 can be used to conduct in vivo, in vitro, or in silico screens for structurally unrelated anti-cancer molecules. For example, competition and other assays known in the art can be used to identify drugs that can outcompete the target interaction with higher affinity and compare the changes in levels to the respective changes caused by a compound of formula (10) or its analogs. Assays can also be performed in live mammalian cells, thereby more closely approximating the in vivo effect of a particular serum level of the drug, or assays can be performed in microsomal extracts prepared from cultured cell lines.

[0136] In one embodiment, the subject has or is at risk of having cancer. In one embodiment, the treatment regimen comprises administering an effective amount of an imipridone or analog thereof, such as ONC201. In one embodiment, the treatment regimen comprises administering an effective amount of ONC201. In one embodiment, the treatment regimen comprises administering an effective amount of a compound of structural formula (10). In one embodiment, the compound of structural formula (10) is a compound of structural formula (40), e.g., a compound of structural formula (45). In one embodiment, the compound of structural formula (10) is a compound of structural formula (50), e.g., a compound of structural formula (55). In one embodiment, the compound of structural formula (10) is a compound of structural formula (80). In one embodiment, the compound of structural formula (10) is a compound of structural formula (90). In one embodiment, the compound of structural formula (10) is a compound of structural 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).

[0137] The predetermined reference level can be, for example, the mean or median of the levels measured in samples from subjects. The predetermined reference level can be measured under the same or substantially similar experimental conditions as those used to measure samples from subjects. The predetermined reference level can be obtained from subjects who respond to treatment with an imipridone such as ONC201 or an analog thereof. In one embodiment, the predetermined reference level is obtained from subjects who respond to treatment with the compound, and if the level in the sample from the subject is similar to that of the reference level, the subject can be classified as a potential responder to treatment. The predetermined reference level can be obtained from subjects who do not respond to treatment with the compound. In one embodiment, the predetermined reference level is obtained from subjects who do not respond to treatment with the compound, and if the level in the sample from the subject is different from that of the predetermined reference level (e.g., up-regulated or down-regulated), the subject can be classified as a potential responder to treatment. The predetermined reference level can also be obtained from normal, healthy subjects.

[0138] Immunoassays, including enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunofiltration assay (ELIFA), flow cytometry, immunoblot, immunoprecipitation, immunohistochemistry, immunocytochemistry, luminescence immunoassay (LIA), fluorescence immunoassay (FIA), and radioimmunoassay, can be used to assay protein or methylation levels in a sample. 6 The level of mRNA methylation can be obtained by methylated RNA immunoprecipitation (Me-RIP) or other quantitative biochemical assays known in the art.

[0139] Nucleic acid mutations can be revealed by any of several known methods. For example, a biological sample can first be collected from an individual. Such biological samples include, but are not limited to, bodily fluids (e.g., urine, saliva, plasma, or serum) or tissue samples (such as oral tissue samples or oral cells). The biological sample can then be sequenced or scanned using known methods. For example, a DNA array can be used to analyze at least a portion of the subject's genome sequence. Furthermore, full or partial genome sequence information can be used. Such sequences can be determined using standard sequencing methods, including chain termination (Sanger dideoxynucleotide), dye terminator sequencing, and SOLID™ sequencing (Applied Biosystems). The entire genome sequence can be digested with restriction enzymes or sheared (mechanically) to shorter fragments for sequencing. DNA sequences can also be amplified using known methods, such as PCR and vector-based cloning (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 thereof) is scanned or sequenced, for example using conventional DNA sequencers or chip-based technology, to reveal the presence or absence of mutations or changes in copy number.

[0140] In one aspect, the present invention provides a method for identifying and treating a subject having a condition who is likely to respond to a treatment regimen described herein. In one embodiment, the method includes (i) determining whether a subject having the condition is likely to respond to a treatment regimen described herein; and (ii) treating the 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 cancer. In one embodiment, the treatment regimen includes administering an effective amount of an imipridone, such as ONC201, or an analog thereof. In one embodiment, the treatment regimen includes administering an effective amount of Compound (1). In one embodiment, the treatment regimen includes administering an effective amount of a compound of structural formula (10). In one embodiment, the compound of structural formula (10) is a compound of structural formula (40), e.g., a compound of structural formula (45). In one embodiment, the compound of structural formula (10) is a compound of structural formula (50), e.g., a compound of structural formula (55). In one embodiment, the compound of structural formula (10) is a compound of structural 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).

[0141] The predetermined reference level can be, for example, the mean or median of the levels measured in samples from subjects. The predetermined reference level can be measured under the same or substantially similar experimental conditions as those used to measure samples from subjects. The predetermined reference level can be obtained from subjects who respond to treatment with an imipridone such as ONC201 or an analog thereof. In one embodiment, the predetermined reference level is obtained from subjects who respond to treatment with the compound, and if the level in the sample from the subject is similar to that of the reference level, the subject can be classified as a potential responder to treatment. The predetermined reference level can be obtained from subjects who do not respond to treatment with the compound. In one embodiment, the predetermined reference level is obtained from subjects who do not respond to treatment with the compound, and if the level in the sample from the subject is different from that of the predetermined reference level (e.g., up-regulated or down-regulated), the subject can be classified as a potential responder to treatment. The predetermined reference level can also be obtained from normal, healthy subjects. Protein levels in samples can be assayed using immunoassays.

[0142] In one aspect, the present invention provides methods for treating a subject having a pathological condition and methods for assessing the effectiveness of the treatment. In one embodiment, the method comprises (i) treating the subject with a treatment method described herein and (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 administering an effective amount of an imipridone, such as ONC201, or an analog thereof. In one embodiment, the treatment regimen comprises administering an effective amount of compound (1). In one embodiment, the treatment regimen comprises administering an effective amount of a compound of structural formula (10). In one embodiment, the compound of structural formula (10) is a compound of structural formula (40), e.g., a compound of structural formula (45). In one embodiment, the compound of structural formula (10) is a compound of structural formula (50), e.g., a compound of structural formula (55). In one embodiment, the compound of structural formula (10) is a compound of structural formula (80). In one embodiment, the compound of structural formula (10) is a compound of structural 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).

[0143] Other conditions that may be amenable to the methods described herein include attention deficit disorder; addiction; epilepsy; viral infections; inflammation; neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, etc.; cardiovascular diseases such as coronary artery disease, cardiomyopathy, hypertensive heart disease, heart failure, pulmonary heart disease, arrhythmias, inflammatory heart disease, endocarditis, inflammatory cardiac hypertrophy, myocarditis, valvular heart disease, cerebrovascular disease, peripheral arterial disease, congenital heart disease, rheumatic heart disease, etc.; diabetes; and light chain amyloidosis.

[0144] V. Composition

[0145] In one aspect, the present invention provides a pharmaceutical composition comprising a compound represented by the following structural formula (10), or a compound represented by the following structural formula (1), or a pharmaceutically acceptable salt thereof:

[0146] [ka]

[0147] [ka]

[0148] In one embodiment, the salt is a pharmaceutically acceptable monosalt of the compound. 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 multiple salt (e.g., di- or tri-salt) selected from the group consisting of hydrochloride, hydrobromide, bisulfate, sulfate, phosphate, fumarate, succinate, oxalate, lactate, bisulfate, hydroxide, tartrate, nitrate, citrate, bitartrate, carbonate, malate, maleate, fumarate, sulfonate, methylsulfonate, 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 having a counterion selected from the group consisting of ammonium, sodium, potassium, calcium, magnesium, zinc, lithium, and / or a counterion such as methylamino, dimethylamino, diethylamino, and triethylamino, hi one embodiment, the salt is a dihydrochloride or dihydrobromide salt.

[0149] Compound (1) (ONC201) has the same chemical structure as revealed by structural analysis (e.g., NMR, X-ray diffraction) of compound NSC350625, available from the Developmental Therapeutics Program Repository of the National Cancer Institute.

[0150] In one embodiment, the pharmaceutical composition comprises a di-salt (e.g., dihydrochloride salt) of ONC201 or an analog thereof (e.g., imipridone). Salts (e.g., di- or tri-salts) of ONC201 analogs can be prepared from ONC201 analogs, which can be synthesized as described herein or using standard chemical synthesis methods known to those skilled in the art.

[0151] In one embodiment, the pharmaceutical composition comprises at least one pharmaceutically acceptable carrier. Suitable pharmaceutically acceptable carriers include, but are not limited to, those in Handbook of Pharmaceutical Excipients, 7th Edition, edited by Raymond C. Rowe et al., American Pharmaceutical Association, Washington, USA and Pharmaceutical Press, London and earlier editions. Exemplary pharmaceutically acceptable carriers, pharmaceutical compositions, and methods of manufacturing various dosage forms and modes of administration are well known in the art, as detailed, for example, in Pharmaceutical Dosage Forms: Tablets, edited by Larry L. Augsburger and Stephen W. Hoag, London: Informa Healthcare, 2008; 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 ed., 2005, particularly chapter 89; and J.G. Hardman et al., Goodman & Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill Professional, 10th ed., 2001.

[0152] In one embodiment, the pharmaceutical composition is formulated for intraocular administration. In one embodiment, the pharmaceutical composition is formulated for topical administration. In one embodiment, the pharmaceutical composition is formulated as drops, ointments, or solutions. In one embodiment, the pharmaceutical composition comprises a conventional pharmaceutical carrier, such as an aqueous, powder, or oily base, a thickener, or the like.

[0153] In one embodiment, the pharmaceutical composition is a formulation for intravenous administration. In one embodiment, the intravenous formulation comprises a compound of Structural 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 a 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 a 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 about 0.005%, 0.05%, or 0.5% to about 0.1%, 1%, or 10% of the compound or a salt thereof. In one embodiment, the intravenous formulation comprises about 0.05%, 0.5%, or 5% of the compound or a salt thereof. In one embodiment, the intravenous formulation comprises a higher or lower concentration of the compound or a salt thereof.

[0154] In one embodiment, the intravenous formulation has a pH of about 3. In one embodiment, the intravenous 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 its salt at a concentration of about 5 mg / ml and a pH of 3, which forms a stable solution. In one embodiment, the intravenous formulation comprises the compound or its salt at a concentration of about 5 mg / ml and a pH of less than 5, which forms a stable solution. In one embodiment, the intravenous formulation comprises the compound or its salt and one or more antioxidants. In one embodiment, the intravenous formulation comprises a mixture of the monohydrochloride and dihydrochloride salts of the compound. In one embodiment, the intravenous formulation comprises the compound or its salt as a 1% solution at a concentration of about 10 mg / mL. For example, the intravenous formulation is a solution with a pH of about 3.3. In one embodiment, the pH is less than 4.0.

[0155] In one embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. In one embodiment, a suitable pharmaceutically acceptable carrier comprises an aqueous carrier. 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.

[0156] 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, a formulation at about 5 mg / ml forms a stable solution and a pH of 3. In one embodiment, a formulation at about 5 mg / ml has a pH of less than 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 monohydrochloride and dihydrochloride salts of ONC201 or an analog thereof. In one embodiment, the intravenous formulation comprises ONC201 or an analog thereof or their 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.

[0157] In one embodiment, the intravenous formulation contains about 0.5% to about 10% (or about 5 mg / ml to about 100 mg / ml) ONC201 or an analog thereof, or a di-salt thereof. In one embodiment, the formulation contains about 5% (or about 50 mg / ml) ONC201 or an analog thereof, or a di-salt thereof. In one embodiment, the intravenous infusion rate may be reduced to reduce side effects of ONC201 or an analog thereof, or a di-salt thereof.

[0158] In one embodiment, the pharmaceutical composition comprises about 0.1-99% of an ONC201 salt or analog thereof and a pharmaceutically acceptable carrier, such as an oil or sterile water or other aqueous carrier. In one embodiment, the pharmaceutical composition comprises about 5% to about 50% of a mono- or disalt of ONC201 or an analog thereof for oral dosage forms.

[0159] In one embodiment, the pharmaceutical composition comprises an antioxidant.Suitable antioxidants include ascorbic acid derivatives, such as ascorbic acid, erythorbic acid, sodium ascorbate, etc., thiol derivatives, such as thioglycerol, cysteine, acetylcysteine, cystine, dithioerythritol, dithiothreitol, glutathione, etc., tocopherol, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), sulfites, such as sodium sulfate, sodium bisulfite, sodium acetone bisulfite, sodium disulfite, sodium sulfite, sodium formaldehyde sulfoxylate and sodium thiosulfate, and nordihydroguaiaretic acid.It should be noted that the antioxidants used in aqueous formulations usually include sodium sulfite, sodium disulfite, sodium formaldehyde sulfoxylate and ascorbic acid and combinations thereof, while the antioxidants used in oily solutions and organic solvents include butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate and combinations thereof. In yet another embodiment, the antioxidant can be one or more of flavanoids, isoflavones, monothioglycerol, L-cysteine, thioglycolic acid, α-tocopherol, ascorbic acid 6-palmitate, dihydrolipoic acid, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), vitamin E, propyl gallate, β-carotene, and ascorbic acid. Antioxidants are typically used at about 0.1% to 1.0% by weight, more typically about 0.2%.

[0160] In one embodiment, the pharmaceutical composition comprises an imipridone or analog thereof, such as ONC201, or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent. For example, the other therapeutic agent may be a hormone analog or antihormonal agent, an aromatase inhibitor, an LHRH agonist or antagonist, a growth factor inhibitor, a growth factor antibody, a growth factor receptor antibody, a tyrosine kinase inhibitor, an antimetabolite, an antitumor antibiotic, a platinum derivative, an alkylating agent, an antimitotic agent, a tubulin inhibitor, a PARP inhibitor, a topoisomerase inhibitor, a serine / threonine kinase inhibitor, a tyrosine kinase inhibitor, a protein-protein interaction inhibitor, a RAF inhibitor, a MEK inhibitor, an ERK inhibitor, an IGF-1R inhibitor, or an IGF-1R inhibitor. anti-inflammatory drugs, ErbB receptor inhibitors, rapamycin analogs, BTK inhibitors, CRM1 inhibitors (e.g., KPT185), P53 modulators (e.g., nutlin), anti-angiogenic agents (e.g., axitinib, aflibercept, sorafenib, and regorafenib), amifostine, anagrelide, clodronate, filgrastin, interferon, interferon alpha, leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate, and pol Fimer, 2-chlorodesoxyadenosine, 2-fluorodesoxy-cytidine, 2-methoxyestradiol, 2C4,3-arretin, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A105972, A204197, abiraterone, aldesleukin, alitretinoin, allovectin-7, altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037, AP-5280, apaziquone, apomine, alano ace, Arglavin, Arzoxifene, Atamestane, Atrasentan, Auristatin PE, ABT-199 (Venetoclax), ABT-263 (Navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (Ganitumab), ARRY162, ARRY438162, 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, azaepothilone B, azonafide, BAY-43-9006, BAY80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, biricodal dicitrate, BCX-1777, BKM-120, bleocin, BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW2992 (afatinib, tom Tobok), BIBF1120 (Valgatef), BI836845, BI2536, BI6727, BI836845, BI847325, BI853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryocistatin-1, bortezomib, brostallicin, busulfan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, canertinib, canfosfamide, capecitabine, carboxyphthalatoplatin, CCl-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, cefratonin, ceftriaxone, celecoxib, celmoleukin, Cemadotin, CH4987655 / RO-4987655, chlorotrianisene, cilengitide, cyclosporine, CDA-II, CDC-394, CKD-602, CKI-27, clofarabine, colchicine, combretastatin A4, COT inhibitors, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibody, CP-461, CV-247 , cyanomorpholinodoxorubicin, cytarabine, D24851, decitabine, deoxorubicin, deoxyrubicin, deoxycoformycin, depsipeptide, desoxyepothilone B, dexamethasone, dexrazoxanet, diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10, doranidazole, DS-7423, E7010, E-6201, edatrexate,Edotreotide, efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509, enzastaurin, enzalutamide, elsamitrucin, epothilone B, epratuzumab, ER-86526, erlotinib, ET-18-0CH3, ethinylcytidine, ethinylestradiol, exatecan, exatecan mesylate, exemestane, exisulind, fenretinide, figitumumab, floxuridine, folic acid, FOLFOX, FOLFOX4, FOLFIRI, formestane, fotemustine, galarubicin, Gallium maltolate, gefinitib, gemtuzumab, gimatecan, glufosfamide, GCS-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, ibandronic acid, ibrutinib, ibritumomab, idatrexate, idenestrol, IDN-5109, IGF-1R inhibitor, IMC-1C11, IMC-A12 (cixutumumab), Immunol, I Indisulam, interferon alpha-2a, interferon alpha-2b, pegylated interferon alpha-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib, ipilimumab, iproplatin, irofulven, isohomohalichondrin-B, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, Kahalid F, ketoconazole, KW-2170, KW-2450, lobaplatin, leflunomide, lenograstim, leuprolide,Leuporelin, lexidronam, LGD-1550, linezolid, lutetium texaphyrin, lometrexol, losoxantrone, LU223651, lurtotecan, LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechloroethamine, MEK inhibitors, MEK-162, methyltestosterone, methylprednisolone, MEDI-573, MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, midostaurin, minodronic acid, mitomycin, Vovulin, MK-2206, MK-0646 (dalotuzumab), MLN518, motexafin gadolinium, MS-209, MS-275, MX6, neridronic acid, neratinib, nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nolatrexed, norelin, N-acetylcysteine, 6-benzylguanine, oblimersen, omeprazole, Oncophage, OncoVEX GM-CSF, olmiplatin, 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, perillyl alcohol, pertuzumab, PI 3K inhibitors, PI3K / mTOR inhibitors, PG-TXL, PG2, PLX-4032 / RO-5185426 (vemurafenib), PLX-3603 / RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pivaloyloxymethyl butyrate, pixantrone, phenoxodiol O, PKI166, previtrexed, plicamycin, polyprenoic acid, porfiromycin, prednisone, prednisolone, quinamed, quinupristin, R115777, RAF-265, ramosetron, ranpirnase,RDEA-119 / BAY869766, RDEA-436, rebeccamycin analogs, receptor tyrosine kinase (RTK) inhibitors, regorafenib, revimid, RG-7167, RG-7304, RG-7421, RG-7321, RG7440, rhizoxin, rhu-MAb, linfavert, risedronate, rituximab, lobatumumab, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR109881A, rubidazone, rubitecan, R-flurbiprofen, RX-0201, S-9788, sabalubicin, SAHA, sargramostim, satraplatin, SB408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustine, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897, SR-3 1747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine, suberanilohydroxamic acid, Sutent, T900607, T138067, TAK-733, TAS-103, tacedinaline, talaporfin, tarceva, tariquitar, tasisulam, taxotere, taxoplexin, tazarotene, tegafur, temozolamide, tesmilifen, testos Telon, testosterone propionate, tesmilifen, tetraplatin, tetrodotoxin, tezacitabine, thalidomide, telarux, terarubicin, thymalfasin, thymectacin, tiazofurin, tipifarnib, tirapazamine, tocladesine, tomudex, tremofin, trabectedin, TransMID-107, transretinoic acid acid), traszutumab, tremelimumab, tretinoin, triacetyluridine, triapine, triciribine, trimetrexate, TLK-286TXD258, Tycarb / Tybarb, Urocidine, valrubicin, vatalanib, vincristine, vinflunine, viridine, 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,Selected from the group consisting of ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZD1839, ZSTK-474, zoledronate, zosuquidar and combinations thereof.

[0161] In one embodiment, the other therapeutic agent comprises a hormone analog, an antihormonal agent, or both 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, the other therapeutic agent comprises one or more LHRH agonists and / or antagonists selected from goserelin acetate, luprolide acetate, triptorelin pamoate, and combinations thereof, wherein the LHRH antagonist is selected from degarelix, cetrorelix, abarelix, ozarelix, degarelix, and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more growth factor inhibitors selected from 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 HER2, HER3, and HER4. In one embodiment, the other therapeutic agent comprises one or more tyrosine kinase inhibitors selected from cetuximab, gefitinib, imatinib, lapatinib, and trastuzumab, and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more aromatase inhibitors selected from anastrozole, letrozole, liarozole, vorozole, exemestane, atamestane, and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more antimetabolites that are antifolates selected from methotrexate, raltitrexed, and pyrimidine analogs. In one embodiment, the other therapeutic agents include one or more antimetabolites that are pyrimidine analogs selected from 5-fluorouracil, capecitabine, and gemcitabine. In one embodiment, the other therapeutic agents include mercaptopurine, thioguanine, cladribine, and pentostatin, cytarabine,The other therapeutic agents include one or more antimetabolites that are purine and / or adenosine analogs selected from fludarabine and combinations thereof. In one embodiment, the other therapeutic agents include one or more antitumor antibiotics selected from anthracyclines, doxorubicin, daunorubicin, epirubicin and idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin, streptozocin, and combinations thereof. In one embodiment, the other therapeutic agents include one or more platinum derivatives selected from cisplatin, oxaliplatin, carboplatin, and combinations thereof. In one embodiment, the other therapeutic agents include one or more alkylating agents selected from estramustine, mechlorethamine, melphalan, chlorambucil, busulfan, dacarbazine, cyclophosphamide, ifosfamide, temozolomide, nitrosoureas, and combinations thereof. In one embodiment, the other therapeutic agents include nitrosoureas selected from carmustine, lomustine, thiotepa, and combinations thereof. In one embodiment, the other therapeutic agent comprises an antimitotic agent selected from a vinca alkaloid and a taxane. In one embodiment, the other therapeutic agent comprises one or more taxanes selected from paclitaxel, docetaxel, and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more vinca alkaloids selected from vinblastine, vindesine, vinorelbine, vincristine, and combinations thereof. In one embodiment, the other therapeutic agent comprises one or more topoisomerase inhibitors that are epipodophyllotoxins. In one embodiment, the other therapeutic agent comprises one or more epipodophyllotoxins selected from etoposide and etopophos, teniposide, amsacrine, topotecan, irinotecan, mitoxantrone, and combinations thereof. In one embodiment, the other therapeutic agents comprise one or more serine / threonine kinase inhibitors selected from a PDK1 inhibitor, a B-Raf inhibitor, an mTOR inhibitor, an mTORC1 inhibitor, a PI3K inhibitor, a dual mTOR / PI3K inhibitor, an STK33 inhibitor, an AKT inhibitor, a PLK1 inhibitor, a CDK inhibitor, an Aurora kinase inhibitor, and combinations thereof.The other therapeutic agents include one or more tyrosine kinase inhibitors that are PTK2 / FAK inhibitors. In one embodiment, the other therapeutic agents include one or more protein-protein interaction inhibitors selected from IAP, Mcl-1, MDM2 / MDMX, and combinations thereof. In one embodiment, the other therapeutic agents include one or more rapamycin analogs selected from everolimus, temsirolimus, ridaforolimus, sirolimus, and combinations thereof. In one embodiment, the other therapeutic agents include one or more therapeutic agents selected from amifostine, anagrelide, clodronate, filgrastin, interferon, interferon alpha, leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate, and porfimer, and combinations thereof. In one embodiment, the other therapeutic agent is 2-chlorodesoxyadenosine, 2-fluorodesoxy-cytidine, 2-methoxyestradiol, 2C4,3-arretin, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A105972, A204197, abiraterone, aldesleukin, alitretinoin, allovectin-7, altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037, AP-5280, apaziquone, apomine, alanose, arglabin, arzoxifene, atamestane, atrasentan, auristatin PE, ABT-199 (venetoclax), ABT-263 (navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), ARRY162, ARRY438162, 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, azaepothilone B, azonafide, BAY-43-9006, BAY80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, bilirubin dicitrate,BCX-1777, BKM-120, bleocin, BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW2992 (afatinib, tomtobok), BIBF1120 (valgatef), BI836845, BI2536, BI6727, BI836845, BI847325, BI853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryocyta Chin-1, bortezomib, brostallicin, busulfan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, canertinib, canfosfamide, capecitabine, carboxyphthalatoplatin, CCl-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1, cefixime, cefratonin, ceftriaxone, celecoxib, celmoleukin, cemadotin, CH4987655 / RO-4987655, chlorotrianisene, cilengitide, cyclosporine, CDA-II, CDC -394, CKD-602, CKI-27, clofarabine, colchicine, combretastatin A4, COT inhibitor, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibody, CP-461, CV-247, cyanomorpholinodoxorubicin, cytarabine, D24851, decitabine, deoxorubicin, deoxyrubicin, deoxycoformycin, depsipeptide, desoxyepothilone B, dexamethasone, dexrazoxane t), diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10, doranidazole, DS-7423, E7010, E-6201, edatrexate, edotreotide, efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509, enzastaurin, enzalutamide, elsamitrucin, epothilone B, epratuzumab, ER-86526, erlotinib, ET-18-0CH3, ethinylcytidine, ethinyl estradiol, exatecan, exatecan mesylate, exemestane,Exisulind, fenretinide, figitumumab, floxuridine, folic acid, FOLFOX, FOLFOX4, FOLFIRI, formestane, fotemustine, galarubicin, gallium maltolate, gefinitib, gemtuzumab, gimatecan, glufosfamide, GCS-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, ibritumomab, idatrexate ate), 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, irofulven, isohomohalichondrin-B, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-10 7088, conjugated estrogens, Kahalid F, ketoconazole, KW-2170, KW-2450, lobaplatin, leflunomide, lenograstim, leuprolide, leuporelin, lexidronam, LGD-1550, linezolid, lutetium texaphyrin, lometrexol, losoxantrone, LU223651, lurtotecan, LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechloroethamine, 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 (dalotuzumab), MLN518, motexafin gadolinium, MS-209, MS-275, MX6, neridronic acid, neratinib, nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nolatrexed, norepinephrine, Phosphorus, N-acetylcysteine, 06-benzylguanine, oblimersen, omeprazole, Oncophage, OncoVEX GM-CSF, olmiplatin, 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-0 05, 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), PLX-3603 / RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pivaloyloxymethyl butyrate, pixantro phenoxodiol O, PKI166, previtrexed, plicamycin, polyprenoic acid, porfiromycin, prednisone, prednisolone, quinamed, quinupristin, R115777, RAF-265, ramosetron, ranpirnase, RDEA-119 / BAY869766, RDEA-436, rebeccamycin analogs, receptor tyrosine kinase (RTK) inhibitors, revimid, RG-7167, RG-7304, RG-7421, RG-7321, RG7440, rhizoxin, rhu-MAb, rinfavert, risedronate, Rituximab Cimab, lobatumumab, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR109881A, rubidazone, rubitecan, R-flurbiprofen, RX-0201, S-9788, sabalubicin, SAHA, sargramostim, satraplatin, SB408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustine, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172,sorafenib, spiroplatin, squalamine, suberanilohydroxamic acid, Sutent, T900607, T138067, TAK-733, TAS-103, tacedinaline, talaporfin, tarceva, tariquitar, tasisulam, taxotere, taxoplexin, tazarotene, tegafur, temozolamide, tesmilifene, testosterone, testosterone propionate, tesmilifene, tetraplatin, tetrodotoxin, tezacitabine, thalidomide, telarux, telarubicin, thymalfasin, thymectacin, tiazofurin, tipifarnib, tirapazamine, tocladesine, tomudex, tremofin, trabectedin, trans-MID-107, trans-retinoic acid acid), traszutumab, tremelimumab, tretinoin, triacetyluridine, triapine, triciribine, trimetrexate, TLK-286TXD258, Tycarb / Tybarb, Urocidine, valrubicin, vatalanib, vincristine, vinflunine, virudin, WX-UK1, WX-554, Vectibix, Xeloda, XELO X, 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, zoledronate, zosuquidar, and combinations thereof.

[0162] In one embodiment, the other therapeutic agent comprises a steroid, including dexamethasone, prednisolone, methylprednisolone, prednisone, hydrocortisone, triamcinolone, betamethasone, and cortivazol. In one embodiment, the other 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, alizapride, prochlorperazine, and metoclopramide), and NK1 receptor antagonists (e.g., aprepitant and casopitant). , antihistamines (such as cyclizine, diphenhydramine, dimenhydrinate, doxylamine, meclizine, promethazine, hydroxyzine, etc.), cannabinoids (such as cannabis, dronabinol, nabilone, and Sativex), benzodiazepines (such as midazolam and lorazepam), anticholinergics (such as hyoscine), trimethobenzamide, ginger, emetrol, propofol, peppermint, muscimol, and ajowan.

[0163] In one embodiment, the other therapeutic agent comprises an anti-cancer agent, which includes an anti-mitotic agent. In one embodiment, the anti-mitotic agent comprises a taxane. In one embodiment, the anti-mitotic agent comprises a taxane selected from paclitaxel and docetaxel.

[0164] In one embodiment, the pharmaceutical composition comprises an imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, and at least one anticancer agent, including acivicin, aclarubicin, acodazole, acronine, adozelesin, aldesleukin, alitretinoin, allopurinol, altretamine, ambomycin, amethanthrone, amifostine, aminoglutethimide, amsacrine, anastrozole, anthramycin, arsenic trioxide, asparaginase, asperlin, azacytidine, azetepa, azotomycin, batimas Tat, benzodepa, bevacizumab, bicalutamide, bisantrene, visnafide dimesylate, bizelesin, bleomycin, brequinar, bropirimine, busulfan, cactinomycin, calsterone, capecitabine, caracemide, carbetimer, carboplatin, carmustine, carubicin, carzelesin, cedefingol, celecoxib, chlorambucil, ciloremycin, cisplatin, cladribine, crisnatol mesylate, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, decitabine, dexol Maplatin, dezaguanine, dezaguanine mesylate, diaziquone, docetaxel, doxorubicin, droloxifene, dromostanolone, duazomycin, edatrexate, eflomithine, elsamitrucin, enloplatin, enpromate, epipropizine, epirubicin, elbrozole, esorubicin, estramustine, etanidazole, etoposide, etopurine, fadrozole, fazarabine, fenretinide, floxuridine, fludarabine, fluorouracil, flurocitabine, foskidone, foskidone Striesin, fulvestrant, gemcitabine, hydroxyurea, idarubicin, ifosfamide, ilmofosine, interleukin II (IL-2, including recombinant interleukin II or rIL2), interferon alpha-2a, interferon alpha-2b, interferon alpha-n1, interferon alpha-n3, interferon beta-Ia, interferon gamma-Ib, iproplatin, irinotecan, lanreotide, letrozole, leuprolide, liarozole, lometrexol, lomustine, losoxantrone, masoprocol,Maytansine, mechlorethamine hydrochloride, megestrol, melengestrol acetate, melphalan, menogaril, mercaptopurine, methotrexate, metoprine, meturedepa, mitindomide, mitocalcine, mitochromine, mitogillin, mitomarcine, mitomycin, mitospel, mitotane, mitoxantrone, mycophenolate, nelarabine, nocodazole, nogalamycin, ormunaplatin, oxisuran, paclitaxel, pegaspargase, periomycin, pentamustine, peplomycin, perfosfamide, pipobroman, piposulfan, piroxantrone hydrochloride, plicamycin, promestane, porfimer, porfiromycin, prednimustine, procarbazine, puromycin, pyrazofurin, ribopurine, rogletimide, safingol, semustine, simtrazene, sparfosate , sparsomycin, spirogermanium, spiromustine, spiroplatin, streptonigrin, streptozocin, sulofenur, tallysomycin, tamoxifen, tecogalan, tegafur, teloxantrone, temoporfin, teniposide, teroxylone, testolactone, thiamiprine, thioguanine, thiotepa, tiazofurin, tirapazamine, topotecan, toremifene, trestron, triciribine, trimetrexate, triptorelin, tubrozole, uracil mustard, uredepa, vapreotide, verteporfin, vinblastine, vincristine sulfate, vindesine, vinepidine, vinglisinate, vinleurosine, vinorelbine, vinrocidine, vinzolidine, vorozole, zeniplatin, zinostatin, zoledronic acid, zorubicin, and combinations thereof.

[0165] Examples of suitable anti-cancer agents include those described in The Pharmacological Basis of Therapeutics, 12th Edition, by Goodman and Gilman, edited by Laurence Brunton, Bruce Chabner, and Bjorn Knollman, McGraw Hill Professional, 2010.

[0166] In some exemplary embodiments, the pharmaceutical composition comprises a salt (e.g., mono- or di-salt) of imipridone, e.g., ONC201, or an analog thereof, and at least one other therapeutic agent, wherein the other therapeutic agent comprises an anti-angiogenic agent, e.g., bevacizumab. In one embodiment, the anti-angiogenic agent is aflibercept, axitinib, angiostatin, endostatin, 16 kDa prolactin fragment, laminin peptide, fibronectin peptide, tissue inhibitor of metalloproteinases (TIMP1, 2, 3, 4), plasminogen activator inhibitor (PAI-1, -2), tumor necrosis factor alpha (high dose, in in vitro), TGF-β1, interferons (IFN-α, -β, -γ), ELR-CXC chemokines, IL-12; SDF-1; MIG; platelet factor 4 (PF-4); IP-10, thrombospondin (TSP), SPARC, 2-methoxyestradiol, proliferin-related protein, suramin, sorafenib, regorafenib, thalidomide, cortisone, linomide, fumagillin (AGM-1470; TNP-470), tamoxifen, retinoids, CM101, dexamethasone, leukemia inhibitory factor (LIF), hedgehog inhibitors, and combinations thereof.

[0167] The pharmaceutical combination can contain the first therapeutic agent and the second therapeutic agent in any desired ratio, so long as a synergistic or cooperative effect still occurs. Preferably, the synergistic pharmaceutical combination contains the first therapeutic agent and the second therapeutic agent in a ratio of about 1:9 to about 9:1. In one embodiment, the synergistic combination contains the first therapeutic agent and the second therapeutic agent 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 about 1:1.

[0168] In one embodiment, the second therapeutic agent is selected from allopurinol, arsenous acid, azacitidine, bortezomib, bevacizumab, capecitabine, carboplatin, celecoxib, chlorambucil, clofarabine, cytarabine, dacarbazine, daunorubicin HCl, docetaxel, doxorubicin HCl, floxuridine, gemcitabine HCl, hydroxyurea, ifosfamide, imatinib mesylate, ixabepilone, lenalidomide, megestrol acetate, methotrexate, mitotane, mitoxantrone HCl, oxaliplatin, paclitaxel, pralatrexate, romidepsin, sorafenib, streptozocin, tamoxifen citrate, topotecan HCl, tretinoin, vandetanib, vismodegib, vorinostat, and combinations thereof.

[0169] 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.

[0170] [Table 2-1]

[0171] [Table 2-2]

[0172] In one embodiment, the second therapeutic agent comprises a drug that targets the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor. In one embodiment, the second therapeutic agent comprises recombinant TRAIL or an 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, mapatumumab, lexatumumab, apomab, and rhApo2L / TRAIL. In one embodiment, the second therapeutic agent comprises an active agent selected from camptothecin, 5-FU, capecitabine, cisplatin, doxorubicin, irinotecan, paclitaxel, cisplatin, bortezomib, BH3I-2, rituximab, radiation, triterpenoids, sorafenib, gemcitabine, HDAC inhibitors, carboplatin, T-101 (a gossypol derivative), ABT-263, ABT-737 and GX-15-070 (obatoclax), vorinostat, cetuximab, panitumumab, bevacizumab, ganitumab, interferon gamma, sorafenib, XIAP antagonists, Bcl-2 antagonists, and Smac mimetics.

[0173] VI. Dose

[0174] In one embodiment, the pharmaceutical composition comprises an imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dose range of about 40, 50, 60, or 100 mg to about 2000 mg; about 4, 5, 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 may be based on the free base form of the compound. In one embodiment, the pharmaceutical composition comprises an imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dose 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; at dosage levels ranging 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 from 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 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 of imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof; or at dosage levels ranging from 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 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 of imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof; 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 at dosage levels ranging from 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 about 100 mg to about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900 mg or 2000 mg of imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof. or at dosage levels ranging from about 1 mg to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 190, or 200 mg. In one embodiment, the pharmaceutical composition comprises about 200 mg to about 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or more of imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, based on the free base form of the compound. 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 at dosage levels ranging from 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.In one embodiment, the pharmaceutical composition comprises imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, at a dosage level ranging from about 400 mg to about 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg; from about 40 mg to about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg based on the free base form of the compound. In one embodiment, the pharmaceutical composition comprises an imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in an amount of about 50 mg to about 60, 70, 80, 90, or 100 mg; about 60 mg to about 70, 80, 90, or 100 mg; about 70 mg to about 80, 90, or 100 mg; about 80 mg to about 90 or 100 mg; about 90 mg to about 100 mg; about 5 mg to about 6, 7, 8, 9, or 10 mg; or about 6 mg about 7, 8, 9, or 10 mg; about 7 mg to about 8, 9, or 10 mg, about 8 mg to about 9 or 10 mg; about 9 mg to about 10 mg; about 0.5 mg to about 0.6, 0.7, 0.8, 0.9, or 1 mg; about 0.6 mg to about 0.7, 0.8, 0.9, or 1 mg; about 0.7 mg to about 0.8, 0.9, or 1 mg, about 0.8 mg to about 0.9, or 1 mg; or about 0.9 mg to about 1 mg.

[0175] In one embodiment, the pharmaceutical composition comprises imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, at a dose ranging from 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 imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, at a dose ranging from about 1, 2, 3, 4, 5, 6, 7, 8, or 9 mg / kg to about 10, 20, 30, or 40 mg / kg; from about 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 mg / kg to about 20, 30, or 40 mg / kg; from about 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 mg / kg to about 30 or 40 mg / kg. g / 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; about 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.10, 3.11, 3.12, 3.13, 3.14, 3.15, 3.16, 3.17, 3.18, 3.19, 3.20, 3.21, 3.22, 3.23, 3.24, 3.25, 3.26, 3.27, 3.28, 3.29, 3.30, 3.31, 3.32, 3.33, 3.34, 3.35, 3.36, 3.37, 3.38, or 3.39 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 7, 2.8 or 2.9 mg / kg to about 3 or 4 mg / kg; or 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. at dose levels ranging from 17, 0.18, or 0.19 mg / kg to about 0.20, 0.30, or 0.40 mg / kg; from 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 0.400 mg / kg; or from 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.

[0176] In one embodiment, the pharmaceutical composition contains about 37.5 mg / m of imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof. 2 ~about 1500mg / m 2 ; approx. 3.75mg / m 2 ~about 150mg / m 2 or about 0.4 mg / m 2 ~about 15mg / m 2In one embodiment, the pharmaceutical composition comprises an imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dose range of 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, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 500, 510, 515, 520, 525, 530, 535, 5 0, 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 45, 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, 6 00, 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, 1 050, 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, 1 420, 1425, 1430, 1435, 1440, 1445, 1450, 1455, 1460, 1465, 1470, 1475, 1480, 1485, 1490, 1495mg / m, 2 ~about 1500mg / m 2 ; approx. 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 ~about 150mg / m 2or 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 2 ~about 15mg / m 2 Contains in doses ranging from.

[0177] VII. Dosage Form

[0178] Pharmaceutical compositions suitable for use in the methods described herein can be formulated into dosage forms 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, to reduce the toxicity of the administered therapeutic agent, the oral dosage unit is divided into multiple smaller doses that are administered to the subject over a predetermined period of time. In one embodiment, the oral dosage unit is administered via a tablet or capsule containing a controlled-release formulation that may include multiple 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, suspension, suppository, tablet, and capsule. In one embodiment, the composition is in an oral dosage form selected from the group consisting of a tablet, caplet, capsule, lozenge, syrup, solution, suspension, and elixir. In one embodiment, the composition is an oral dosage form selected from the group consisting of tablets, hard shell and gelatin capsules, beads, granules, agglomerates, powders, gels, solids, and semi-solids.

[0179] In one embodiment, the pharmaceutical composition suitable for use in the methods described herein may comprise a dermatological composition suitable for topical application to the skin.For example, the dermatological composition comprises a cosmetically acceptable or pharmaceutically acceptable medium.The dermatological composition for topical application may comprise an ointment, lotion, cream, gel, drops, suppository, spray, liquid and powder.In one embodiment, conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners, skin enhancers may be used if necessary or desired. Examples of suitable enhancers include ethers such as diethylene glycol monoethyl ether (commercially available as TRANSCUTOL®) and diethylene glycol monomethyl ether; surfactants such as sodium laurate, sodium lauryl sulfate, cetyltrimethylammonium bromide, benzalkonium chloride, poloxamers (231, 182, 184), Tween® (20, 40, 60, 80), and lecithin (U.S. Pat. No. 4,783,450); alcohols such as ethanol, propanol, octanol, benzyl alcohol, polyethylene glycol and its esters, such as polyethylene glycol monolaurate; amides and other nitrogen 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. AZONE® and sulfoxides such as DMSO and C10MSO may also be used, but are less preferred.

[0180] In one embodiment, the pharmaceutical composition is in a dosage form selected from the group consisting of sustained release, controlled release, delayed release and responsive release forms.

[0181] VIII.How to use

[0182] The compositions and methods described herein are useful for treating a variety of disease states, including cancer (e.g., colorectal cancer, brain cancer, and glioblastoma).In one embodiment, the compositions and methods described herein are used to treat diseases such as intraocular melanoma, desmoplastic round cell tumor, chondrosarcoma, leptomeningeal disease, diffuse large B-cell lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancer, 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 tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, and spinal tumor. In one embodiment, the compositions and methods described herein are used to treat diseases such as carcinoid tumors, cancer of unknown primary, central nervous system atypical teratoid / rhabdoid tumor, leptomeningeal disease, central nervous system embryonal tumors, central nervous system lymphoma, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, 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 central nervous system embryonal tumors, endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, Ewing's sarcoma tumor family, 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 tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, and glioma. In one embodiment, the compositions and methods described herein are used to treat a cancer selected from the group consisting of hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, histiocytosis, Hodgkin's lymphoma, and hypopharyngeal carcinoma. In one embodiment, the compositions and methods described herein are used to treat diseases such as Kaposi's sarcoma and kidney (renal cell) carcinoma.In one embodiment, the compositions and methods described herein are used to treat diseases such as Langerhans cell histiocytosis, laryngeal cancer, lip and oral cavity cancer, liver cancer, lung cancer, including non-small cell lung cancer and small cell lung cancer, non-Hodgkin's lymphoma, and primary central nervous system lymphoma. In one embodiment, the compositions and methods described herein are used to treat diseases such as Waldenstrom's macroglobulinemia (lymphoplasmacytic lymphoma), malignant fibrous histiocytoma and osteosarcoma of bone, medulloblastoma, medulloepithelioma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous cell carcinoma of the neck with unknown primary, multiple endocrine neoplasia syndrome, oral cancer, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, complex karyotype blast phase leukemia, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, and myeloproliferative disorders. 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 cancer, parathyroid cancer, penile cancer, pharyngeal cancer, intermediate pineal parenchymal tumor, pineoblastoma and supratentorial primitive neuroectodermal tumor, 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 cancer selected from the group consisting of rectal cancer, renal cell (kidney) cancer, renal pelvis and ureter, airway cancer involving the NUT gene on chromosome 15, retinoblastoma, and rhabdomyosarcoma. In one embodiment, the compositions and methods described herein are used to treat aggressive 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 indolent 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 nervous system tumors. In one embodiment, the compositions and methods described herein are used to treat central nervous system tumors. In one embodiment, the compositions and methods described herein are used to treat peripheral nervous system tumors. In one embodiment, the compositions and methods described herein are used to treat paragangliomas. In one embodiment, the compositions and methods described herein are used to treat pheochromocytoma.

[0183] In vitro models, animal models, and human clinical trials have shown that compound (1) (ONC201) has broad-spectrum anticancer activity and possesses low toxicity with few, if any, adverse effects, low genotoxicity, and high bioavailability, including oral bioavailability. 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 ensuring durable responses or stable disease or preventing disease recurrence in high-risk patients.

[0184] In one embodiment, the compositions and methods described herein are used to treat pediatric cancers (e.g., pediatric solid tumors, pediatric sarcoma, pediatric Ewing's sarcoma, pediatric glioma, pediatric central nervous system cancer, pediatric neuroblastoma, pediatric leukemia, and pediatric lymphoma).

[0185] In one embodiment, the compositions and methods described herein are used to treat proliferative skin disorders 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 cancer, sarcoma, Sézary syndrome, skin cancer, eye cancer, skin cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, cervical squamous cell carcinoma of unknown primary site, 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 carcinoma of the renal pelvis and ureter, and gestational trophoblastic tumor. 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, cancer of unknown primary site, rare cancers of childhood, transitional cell carcinoma 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 women's cancer.

[0186] In one embodiment, the compositions and methods described herein are used as first-line therapy (sometimes referred to as primary therapy). In one embodiment, the compositions and methods described herein are used as second-line therapy. In one embodiment, the compositions and methods described herein are used as third-line therapy. In one embodiment, the compositions and methods described herein are used as salvage therapy. The term "salvage therapy" refers to a therapeutic agent that may be employed in any treatment regimen after a subject's initial treatment regimen has failed or after the subject's condition has not responded to the initial treatment. In one embodiment, the compositions and methods described herein are used as rescue therapy. In one embodiment of rescue therapy, the composition is used as a rescue agent to counteract the effects of the initial treatment. In one embodiment of rescue therapy, the composition is used as a rescue agent administered to a subject who has developed resistance to the standard or initial treatment. In one embodiment, the compositions and methods described herein are used as neoadjuvant therapy. In one embodiment, neoadjuvant therapy involves administering one or more therapeutic agents described herein to a subject prior to the main or first-line treatment. In one embodiment, neoadjuvant therapy reduces the size or extent of the cancer being treated before the main or first-line treatment is administered to the subject being treated. In one embodiment, the compositions and methods described herein are used as adjuvant therapy. In one embodiment, adjuvant therapy involves the administration of one or more therapeutic agents described herein to a subject, where the one or more therapeutic agents alter the effect of other therapeutic agents already administered to the subject, administered simultaneously with the subject, or administered later to the subject.

[0187] In one embodiment, the compositions and methods described herein reduce the potential for drug-drug interactions. In one embodiment, the imipridone, such as ONC201, or its analog, is cleared from the patient's body before it can interact with another pharmaceutically active agent.

[0188] In one embodiment, the compositions and methods described herein exhibit a level of toxicity that is amenable to combination with other pharmaceutical agents.

[0189] The methods and compositions described herein are not limited to any particular animal species. In one embodiment, the subject treated with the compositions described herein according to the methods of the present invention can be a mammal or a non-mammal. In one embodiment, mammalian subjects include, but are not limited to, humans; non-human primates; rodents such as mice, rats, or guinea pigs; domesticated pets such as cats or dogs; horses, cows, pigs, sheep, goats, or rabbits. In one embodiment, non-mammalian subjects include, but are not limited to, birds such as ducks, geese, chickens, or turkeys. In one embodiment, the subject is a human. In one embodiment, the subject can be of either gender 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.

[0190] The compositions and methods described herein are not limited to subjects of a particular age.In one embodiment, the subject treated with the compositions described herein according to the methods described herein is over 50 years old, over 55 years old, over 60 years old, or over 65 years old.In one embodiment, the subject treated with the compositions described herein according to the methods described herein is under 50 years old, under 55 years old, under 60 years old, or under 65 years old.

[0191] In one embodiment, the subject treated with the compositions described herein according to the methods described herein is a pediatric patient. In one embodiment, the pediatric patient is under 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 year old. In one embodiment, the pediatric patient is under 12 months, 11 months, 10 months, 9 months, 8 months, 7, 6 months, 5 months, 4, 3, 2, or 1 month old. In one embodiment, the pediatric patient is under 4 weeks, 3 weeks, 2 weeks, or 1 week old. In one embodiment, the pediatric patient is under 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day old. In one embodiment, the pediatric patient is a newborn. In one embodiment, the pediatric patient is a premature infant.

[0192] In one embodiment, the patient weighs less than 45 kg, weighs less than 40 kg, weighs less than 35 kg, weighs less than 30 kg, weighs less than 25 kg, weighs less than 20 kg, weighs less than 15 kg, weighs less than 14 kg, weighs less than 10 kg, weighs less than 5 kg, weighs less than 4 kg, weighs less than 3 kg, weighs less than 2 kg or weighs less than 1 kg.

[0193] 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.

[0194] In one embodiment, the subject is undergoing radiation therapy, in one embodiment, the subject is undergoing surgery, in one embodiment, the subject is undergoing adoptive T cell therapy.

[0195] In one embodiment, the cancer has stopped responding to treatment with ibrutinib, bortezomib, carfilzomib, temozolomide, bevacizumab, cyclophosphamide, hydroxydaunorubicin, vincristine, prednisone, cytarabine, cisplatin, rituximab, 5-fluorouracil, oxaliplatin, leucovorin, lenalidomide, radiation, surgery, or a combination thereof.

[0196] In one embodiment, the compositions and methods described herein exhibit a dose-response relationship in cancer cells that differs from the dose-response relationship in normal cells. The dose-response relationship of ONC201 on proliferation and cell death of normal and tumor cells was determined by measuring cell viability after 72 hours of treatment with various concentrations of ONC201. The tumors tested included a human colon cancer cell line (HCT116), a breast tumor cell line (MDA-MB-231), and a human primary glioblastoma cell line (U87). The normal cells tested included human foreskin fibroblasts (HFF), human fetal lung fibroblast (MRC-5), and a human lung fibroblast cell line (WI-38). 1 μg / mL doxorubicin was used as a positive control for normal fibroblasts. Cell viability of the normal cells tested was at least approximately 75% at concentrations of 1-5 mg / mL of ONC201, whereas tumor cell viability was significantly lower (e.g., below 50%) at the same ONC201 concentrations. Furthermore, increasing the ONC201 concentration above approximately 5 mg / mL reduced tumor cell viability to less than 25%, while normal cell viability remained at approximately 75%. Cell viability assays of human fetal lung fibroblast (MRC-5) cells were performed after 72 hours of treatment with compound (1) (5 μM) or DMSO, followed by a post-treatment recovery period in complete drug-free medium. ONC201, but not DMSO, resulted in cell recovery.

[0197] In one embodiment, the compositions and methods described herein are useful for treating cancer in a subject. In one embodiment, the compositions and methods described herein are useful for 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 an imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0198] In one embodiment, a method of treatment comprises administering to a subject in need of such treatment (i) a first therapeutic agent comprising an imipridone such as 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 pharmaceutically active agent disclosed herein. Pharmaceutically acceptable salts of ONC201 include the dihydrochloride salt shown below.

[0199] [ka]

[0200] It is understood that in the compositions or dosing regimens described herein, the dihydrochloride salt of ONC201 or an analog thereof (including the compound of structural formula (10)), or an alternative disalt thereto apparent from the teachings of the present disclosure, may be substituted for ONC201 or an analog thereof.

[0201] In one embodiment, a method of treatment comprises administering to a subject in need of such treatment a synergistic combination pharmaceutical formulation, either simultaneously or sequentially, the synergistic combination pharmaceutical formulation comprising (i) a first therapeutic agent comprising an imipridone, such as ONC201, or an analog thereof, or a pharmaceutically acceptable salt thereof, and (ii) a second therapeutic agent. In one embodiment, a method of treatment comprises administering to a subject in need of such treatment a therapeutically synergistic amount of a first therapeutic agent in combination with a second therapeutic agent, either simultaneously or sequentially. In one embodiment, a 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, the combination providing a synergistic effect in the in vivo treatment of a susceptible cancer, the first and second therapeutic agents being administered simultaneously or sequentially. In one embodiment, a 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, the combination providing a synergistic effect in the in vivo treatment of a susceptible minimal residual disease, the first and second therapeutic agents being administered simultaneously or sequentially. In one embodiment, the second therapeutic agent is administered before or prior to the first therapeutic agent.

[0202] 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.

[0203] In one embodiment, the therapeutic method targets solid tumors, including cervical cancer, endometrial cancer, extracranial germ cell tumors; extragonadal germ cell tumors; germ cell tumors; gestational trophoblastic tumors; ovarian cancer, ovarian germ cell tumors, ovarian epithelial cancer, ovarian low-grade malignant tumors; penile cancer, prostate cancer; pregnancy and breast cancer; high-grade prostate cancer; intermediate-grade prostate cancer; low-grade prostate cancer; castration-resistant prostate cancer; breast cancer; cholangiocarcinoma; extrahepatic cholangiocarcinoma; gallbladder cancer; hepatocellular (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, papilloma, multiple endocrine neoplasia syndrome; pancreatic cancer, parathyroid carcinoma, intraocular melanoma; eye cancer; retinoblastoma; malignant fibrous histiocytoma; Ewing's sarcoma tumor family; desmoplastic round cell tumor; chondrosarcoma, Kaposi's sarcoma, rhabdomyosarcoma; spinal cord tumors, leptomeningeal disease, central nervous system embryonal tumors, chordoma, central nervous system embryonal tumors, Ependymoblastoma, ependymoma, neuroblastoma; intermediate pineal parenchymal tumor, pineoblastoma; adrenocortical carcinoma; bone cancer, osteosarcoma; malignant fibrous histiocytoma and osteosarcoma of bone; osteosarcoma and malignant fibrous histiocytoma of bone; carcinoid tumor, cancer of unknown primary, bronchial tumor, lung cancer, pleuropulmonary blastoma; respiratory tract cancer involving the NUT gene on chromosome 15, astrocytoma, atypical teratoid / rhabdoid tumor; central nervous system atypical teratoid / rhabdoid tumor, craniopharyngioma, glioma, brain tumor , medulloblastoma, medulloepithelioma, supratentorial primitive neuroectodermal tumor; pituitary tumor; gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), bladder cancer, anal or rectal cancer, appendix cancer, esophageal cancer, hypopharyngeal cancer; laryngeal cancer, lip and oral cavity cancer, metastatic squamous cell carcinoma of the neck of unknown primary, oral cancer, nasal cavity and paranasal cavity cancer, nasopharyngeal cancer, oral cancer, lip and oral cavity cancer, oropharynx cancer, paranasal cavity and nasal cavity cancer, pharyngeal cancer; head and neck cancer and mesothelioma.

[0204] In one embodiment, the therapeutic method targets a lymphoma selected from the group consisting of diffuse large B-cell lymphoma, AIDS-related lymphoma, cutaneous T-cell lymphoma, Sezary syndrome, mycosis fungoides (MF); histiocytosis; Burkitt's lymphoma and central nervous system lymphoma; non-Hodgkin's lymphoma and primary central nervous system lymphoma, Hodgkin's lymphoma, Waldenstrom's macroglobulinemia; mycosis fungoides; primary central nervous system lymphoma; lymphoplasmacytic lymphoma and primary central nervous system lymphoma.

[0205] In one embodiment, the treatment method targets a non-Hodgkin's lymphoma (NHL) selected from the group consisting of mantle cell lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, marginal zone lymphoma, small lymphocytic lymphoma, lymphoplasmacytic NHL, Waldenstrom's macroglobulinemia, and cutaneous lymphoma.

[0206] In one embodiment, the therapeutic method targets a leukemia selected from the group consisting of acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloproliferative disease; hairy cell leukemia; acute myeloid leukemia (AML); chronic myeloid leukemia (CML); and Langerhans cell histiocytosis.

[0207] In one embodiment, the method of treatment targets an acute leukemia selected from the group consisting of acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphoblastic leukemia, chronic myelogenous leukemia, myelodysplastic syndromes, and myeloproliferative disorders.

[0208] In one embodiment, the therapeutic method 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 blast phase leukemia; multiple myeloma / plasma cell neoplasm, multiple myeloma, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, and myeloproliferative disease.

[0209] In one embodiment, the therapeutic method targets a peripheral nervous system tumor. In one embodiment, the therapeutic method targets a paraganglioma. In one embodiment, the therapeutic method targets a pheochromocytoma.

[0210] In one embodiment, treating cancer comprises preventing tumor growth in a subject with cancer. In one embodiment, treating cancer comprises preventing the formation of cancer metastases in a subject with cancer. In one embodiment, treating cancer comprises targeted treatment of minimal residual disease in a subject with cancer known to have minimal residual disease or at risk of having minimal residual disease in cancer.

[0211] This may be indicated after treatment of the primary tumor by surgery and / or after chemotherapy (radiotherapy) has been initiated or has been determined to be effective. Disseminated tumor cells may be in a dormant state and are often unable to be attacked by chemotherapy (radiotherapy). Patients treated in this manner are apparently cured, a state also known as "minimal residual disease." Nevertheless, dormant tumor cells may form metastases if, after a longer period of dormancy, they become metastatic cells upon proliferative stimulation.

[0212] The term "minimal residual disease" refers to a small number of cancer cells that persist in a subject during or after treatment when the subject is in remission (exhibiting no symptoms or signs of the disease). The methods described herein are preferably applied to the forms of the disease listed herein, including adult and pediatric forms of the disease.

[0213] In one embodiment, the method of treatment is useful for treating autoimmune diseases, including, but not limited to, alopecia areata, antiphospholipid, autoimmune hepatitis, celiac disease, type 1 diabetes, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, hemolytic anemia, idiopathic thrombocytopenic purpura, inflammatory bowel disease, inflammatory myopathy, multiple sclerosis, primary biliary cirrhosis, psoriasis, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus, and vitiligo.

[0214] In one embodiment, the method of treatment is useful for treating autoimmune and inflammatory disorders of the peripheral nervous system, such as metabolic disorders including diabetes and B12 and vitamin folate deficiency, chemotherapeutic agents and drugs used to treat HIV, toxins that cause peripheral nerve damage, cancers that cause peripheral neuropathy and paraneoplastic syndromes, alcohol abuse, chronic kidney disease, injuries that cause nerve compression and other lesions, infectious diseases such as Lyme disease, certain inflammatory conditions such as Guillain-Barré syndrome, connective tissue diseases, rheumatoid arthritis, Sjogren's syndrome, systemic lupus erythematosus, sarcoidosis, genetic disorders such as celiac disease, Charcot-Marie-Tooth syndrome, Friedreich's ataxia, and / or amyotrophic lateral sclerosis (Lou Gehrig's disease) of various etiologies, including idiopathic forms where the specific cause is unknown but where inflammatory and / or autoimmune mechanisms are responsible for their development.

[0215] In one embodiment, the method of treatment is useful for treating autoimmune and inflammatory disorders that manifest with ocular symptoms, including, but not limited to, ocular cicatricial pemphigoid, Mooren's corneal ulcer, various forms of uveitis, rheumatoid arthritis, systemic lupus erythematosus, polyarteritis nodosa, relapsing polychondritis, Wegener's granulomatosis, scleroderma, Behçet's disease, Reiter's disease, inflammatory bowel disease (ulcerative colitis and Crohn's disease), and ankylosing spondylitis, retinitis pigmentosa, macular degeneration, keratoconjunctivitis sicca, scleritis, episcleritis, keratitis, peripheral corneal ulcer, and less common entities such as choroiditis, retinal vasculitis, episcleral nodules, retinal detachment, and / or macular edema.

[0216] In one embodiment, the method of treatment is useful for treating acute allograft rejection in transplant patients. In one embodiment, the method of treatment is useful for treating ischemic stroke. In one embodiment, the method of treatment is useful for treating inflammatory diseases including arthritis, psoriasis, asthma, and colitis.

[0217] In one embodiment, the therapeutic agent comprises a pharmaceutically acceptable monosalt of ONC201 or an analog thereof (e.g., a compound of structural formula (10)). In one embodiment, the therapeutic agent comprises a pharmaceutically acceptable di-salt of ONC201 or an analog thereof (e.g., a compound of structural formula (10)). As described herein, some analogs may be tri-salts. In one embodiment, the therapeutic agent comprises ONC201 or an analog thereof (e.g., a compound of structural formula (10)) in the form of a pharmaceutically acceptable mono- or di-salt selected from the group consisting of hydrochloride, hydrobromide, bisulfate, sulfate, phosphate, fumarate, succinate, oxalate, and lactate, bisulfate, hydroxide, tartrate, nitrate, citrate, bitartrate, carbonate, malate, maleate, fumarate sulfonate, methylsulfonate, formate, acetate, and carboxylate. In one embodiment, the therapeutic agent comprises ONC201 or an analog thereof in the form of a pharmaceutically acceptable mono- or di-salt selected from p-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 having a counterion selected from the group consisting of ammonium, sodium, potassium, calcium, magnesium, zinc, and lithium, and / or having a counterion such as methylamino, dimethylamino, diethylamino, triethylamino, and combinations thereof. In one embodiment, the therapeutic agent comprises a compound described herein in the form of a halide disalt, such as a dihydrochloride or hydrobromide salt.

[0218] 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 comprises acivicin, aclarubicin, acodazole, acronine, adozelesin, aldesleukin, alitretinoin, allopurinol, altretamine, ambomycin, amethanthrone, amifostine, aminoglutethimide, amsacrine, anastrozole, anthramycin, arsenic trioxide, asparaginase, asperlin, azacytidine, azetepa, azotomycin, batimastat, benzodepa, bevacizumab, bicalutamide, bisantrene, bisnafide dimesylate, or bizelesin. , bleomycin, brequinar, bropirimine, busulfan, cactinomycin, calsterone, capecitabine, caracemide, carbetimer, carboplatin, carmustine, carubicin, carzelesin, cedefingol, celecoxib, chlorambucil, ciloremycin, cisplatin, cladribine, crisnatol mesylate, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, decitabine, dexorumaplatin, dezaguanine, dezaguanine mesylate, diaziquone, docetaxel, doxorubicin Rubicin, droloxifene, dromostanolone, duazomycin, edatrexate, eflomithine, elsamitrucin, enloplatin, enpromate, epipropidine, epirubicin, elbrozole, esorubicin, estramustine, etanidazole, etoposide, etopurine, fadrozole, fazarabine, fenretinide, floxuridine, fludarabine, fluorouracil, flurocitabine, foskidone, fostriecin, fulvestrant, gemcitabine, hydroxyurea, ibuprofen Darubicin, ifosfamide, ilmofosine, interleukin II (IL-2, including recombinant interleukin II or rIL2), interferon alpha-2a, interferon alpha-2b, interferon alpha-n1, interferon alpha-n3, interferon beta-Ia, interferon gamma-Ib, iproplatin, irinotecan, lanreotide, letrozole, leuprolide, liarozole, lometrexol, lomustine, losoxantrone, masoprocol, maytansine, mechlorethamine hydrochloride, megestrol,Melengestrol acetate, melphalan, menogaril, mercaptopurine, methotrexate, metoprine, meturedepa, mitindomide, mitocalcin, mitochromine, mitogillin, mitomarcine, mitomycin, mitospel, mitotane, mitoxantrone, mycophenolate, nelarabine, nocodazole, nogalamycin, ormunaplatin, oxisuran, paclitaxel, pegaspargase, periomycin, pentamustine, peplomycin, perfosfamide, pipobroman, piposulfan, piroxantrone hydrochloride, plicamycin, promestane, porfimer, porfiromycin, prednimustine, procarbazine, puromycin, pyrazofurin, ribopurine, rogletimide, safingol, semustine, simtrazene, sparfosate, sparsomycin The anticoagulant is selected from the group consisting of vincristine, spirogermanium, spiromustine, spiroplatin, streptonigrin, streptozocin, sulofenur, tallysomycin, tamoxifen, tecogalan, tegafur, teloxantrone, temoporfin, teniposide, teroxylone, testolactone, thiamiprine, thioguanine, thiotepa, tiazofurin, tirapazamine, topotecan, toremifene, trestron, triciribine, trimetrexate, triptorelin, tubrozole, uracil mustard, uredepa, vapreotide, verteporfin, vinblastine, vincristine sulfate, vindesine, vinepidine, vinglisinate, vinleurosine, vinorelbine, vinrocidine, vinzolidine, vorozole, zeniplatin, zinostatin, zoledronic acid, zorubicin, and combinations thereof.

[0219] In one embodiment of the method of treatment, the second therapeutic agent is selected from the group consisting of hormone analogs and antihormones, 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; 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, and the like. Isin analogues, amifostine, anagrelide, clodronate, filgrastin, interferon, interferon alpha, leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate and porfimer, 2-chlorodesoxyadenosine, 2-fluorodesoxy-cytidine, 2-methoxyestradiol, 2C4,3-arretin, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A105972, A20419 7, abiraterone, aldesleukin, alitretinoin, allovectin-7, altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037, AP-5280, apaziquone, apomine, alanose, arglabin, arzoxifene, atamestane, atrasentan, auristatin PE, ABT-199 (venetoclax), ABT-263 (navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), ARRY162, ARRY438162, ARRY-300, ARRY-14 2886 / 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, azaepothilone B, azonafide, BAY-43-9006, BAY80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, bilirubin dicitrate,BCX-1777, BKM-120, bleocin, BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW2992 (afatinib, tomtobok), BIBF1120 (valgatef), BI836845, BI2536, BI6727, BI836845, BI847325, BI853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryocyta Chin-1, bortezomib, brostallicin, busulfan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, canertinib, canfosfamide, capecitabine, carboxyphthalatoplatin, CCl-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1, cefixime, cefratonin, ceftriaxone, celecoxib, celmoleukin, cemadotin, CH4987655 / RO-4987655, chlorotrianisene, cilengitide, cyclosporine, CDA-II, CDC -394, CKD-602, CKI-27, clofarabine, colchicine, combretastatin A4, COT inhibitor, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibody, CP-461, CV-247, cyanomorpholinodoxorubicin, cytarabine, D24851, decitabine, deoxorubicin, deoxyrubicin, deoxycoformycin, depsipeptide, desoxyepothilone B, dexamethasone, dexrazoxane t), diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10, doranidazole, DS-7423, E7010, E-6201, edatrexate, edotreotide, efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509, enzastaurin, enzalutamide, elsamitrucin, epothilone B, epratuzumab, ER-86526, erlotinib, ET-18-0CH3, ethinylcytidine, ethinyl estradiol, exatecan, exatecan mesylate, exemestane,Exisulind, fenretinide, figitumumab, floxuridine, folic acid, FOLFOX, FOLFOX4, FOLFIRI, formestane, fotemustine, galarubicin, gallium maltolate, gefinitib, gemtuzumab, gimatecan, glufosfamide, GCS-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100, gp100-peptide vaccine, G SK-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, idatrexate, idene 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, irofulven, isohomohalichondrin-B, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugates Gate estrogen, Kahalid F, ketoconazole, KW-2170, KW-2450, lobaplatin, leflunomide, lenograstim, leuprolide, leuporelin, lexidronam, LGD-1550, linezolid, lutetium texaphyrin, lometrexol, losoxantrone, LU223651, lurtotecan, LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechloroethamine, 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, norelin, N-acetylcysteine, 6-benzylguanine, oblimersen, omeprazole, Oncophage, OncoVEXGM -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, P 54, PI-88, pelitinib, 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, pivaloyloxymethyl butyrate, pixantrone, phenoxodiol O, PKI166, previtrexed, plicamycin, polyprenoic acid, porfiromycin, prednisone, Prednisolone, Quinamed, Quinupristin, R115777, RAF-265, Ramosetron, Ranpirnase, RDEA-119 / BAY869766, RDEA-436, Rebeccamycin analogs, Receptor tyrosine kinase (RTK) inhibitors, Revimid, RG-7167, RG-7304, RG-7421, RG-7321, RG7440, Rhizoxin, rhu-MAb, Linfavert, Risedronic acid, Rituximab, Lobatumumab, Rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR109881A,Rubidazone, rubitecan, R-flurbiprofen, RX-0201, S-9788, sabalubicin, SAHA, sargramostim, satraplatin, SB408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustine, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine, suberanilohydroxamic acid, Sutent, T900607, T138067, TAK-733, TAS-103, tacedinaline, tara Porfin, Tarceva, Tariquitar, Tasisulam, Taxotere, Taxoplexin, Tazarotene, Tegafur, Temozolamide, Tesmilifene, Testosterone, Testosterone propionate, Tesmilifene, Tetraplatin, Tetrodotoxin, Tezacitabine, Thalidomide, Telarux, Terarubicin, Thymalfasin, Timectacin, Tiazofurin, Tipifarnib, Tirapazamine, Tocladesine, Tomudex, Tremofin, Trabectedin, TransMID-107, Transretinoic acid acid), traszutumab, tremelimumab, tretinoin, triacetyluridine, triapine, triciribine, trimetrexate, TLK-286TXD258, Tycarb / Tybarb, Urocidin, valrubicin, vatalanib, vincristine, vinflunine, virudin, WX-UK1, WX-554, Vectibix, Zero The steroid inhibitor is selected from the group consisting of tetracycline, 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, zoledronate, zosuquidar, and combinations thereof.

[0220] 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 an LHRH agonist and an LHRH antagonist. In one embodiment, the LHRH agonist is selected from goserelin acetate, luprolide 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 a growth factor inhibitor. In one embodiment, the growth factor inhibitor is selected from 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), hepatocyte growth factor (HGF), and combinations thereof. In one embodiment, the human epidermal growth factor (HER) is selected from HER2, HER3, and HER4.

[0221] 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 tyrosine kinase inhibitor is selected from cetuximab, gefitinib, imatinib, lapatinib, and trastuzumab, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises an aromatase inhibitor. In one embodiment of the method of treatment, the aromatase inhibitor is selected from anastrozole, letrozole, liarozole, vorozole, exemestane, atamestane, and combinations thereof.

[0222] In one embodiment of the method of treatment, the second therapeutic agent comprises an antimetabolite. In one embodiment of the method of treatment, the antimetabolite comprises an antifolate. In one embodiment of the method of treatment, the antifolate is selected from methotrexate, raltitrexed, a pyrimidine analog, and combinations thereof. In one embodiment of the method of treatment, the antimetabolite is a pyrimidine analog. In one embodiment of the method of treatment, the pyrimidine analog is selected from 5-fluorouracil, capecitabine, gemcitabine, and combinations thereof. In one embodiment of the method of treatment, the antimetabolite is a purine analog or an adenosine analog. In one embodiment of the method of treatment, the purine analog or adenosine analog is selected from mercaptopurine, thioguanine, cladribine, pentostatin, cytarabine, fludarabine, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises an antitumor antibiotic. In one embodiment of the method of treatment, the antitumor antibiotic is selected from anthracyclines, doxorubicin, daunorubicin, epirubicin and idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin, streptozocin, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises a platinum derivative. In one embodiment of the method of treatment, the platinum derivative is selected from cisplatin, oxaliplatin, carboplatin, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises an alkylating agent. In one embodiment of the method of treatment, the alkylating agent is selected from estramustine, mechlorethamine, melphalan, chlorambucil, busulfan, dacarbazine, cyclophosphamide, ifosfamide, temozolomide, nitrosoureas, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises a nitrosourea. In one embodiment of the method of treatment, the nitrosourea is selected from carmustine, lomustine, thiotepa, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises an antimitotic agent. In one embodiment of the method of treatment, the antimitotic agent is selected from a vinca alkaloid and a taxane. In one embodiment of the method of treatment, the taxane is selected from paclitaxel, docetaxel, and combinations thereof. In one embodiment of the method of treatment, the vinca alkaloid is vinblastine, vindesine, vinorelbine,vincristine, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises a topoisomerase inhibitor. In one embodiment of the method of treatment, the topoisomerase inhibitor is an epipodophyllotoxin. In one embodiment of the method of treatment, the topoisomerase inhibitor that is an epipodophyllotoxin is selected from etoposide, etopophos, teniposide, amsacrine, topotecan, irinotecan, mitoxantrone, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent comprises a serine / threonine kinase inhibitor. In one embodiment of the method of treatment, the serine / threonine kinase inhibitor is selected from a PDK1 inhibitor, a B-Raf inhibitor, an mTOR inhibitor, an mTORC1 inhibitor, a PI3K inhibitor, an mTOR / PI3K dual inhibitor, an STK33 inhibitor, an AKT inhibitor, a PLK1 inhibitor, a CDK inhibitor, an Aurora kinase inhibitor, 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 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 everolimus, temsirolimus, ridaforolimus, sirolimus, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent is selected from amifostine, anagrelide, clodronate, filgrastin, interferon, interferon alpha, leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate, and porfimer, and combinations thereof. In one embodiment of the method of treatment, the second therapeutic agent is 2-chlorodesoxyadenosine, 2-fluorodesoxy-cytidine, 2-methoxyestradiol, 2C4,3-arretin, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A105972, A204197, abiraterone, aldesleukin, alitretinoin, allovectin-7,Altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037, AP-5280, apaziquone, apomine, alanose, arglabin, arzoxifene, atamestane, atrasentan, auristatin PE, ABT-199 (venetoclax), ABT-263 (nabitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), ARRY162, ARRY438162, 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, azaepothilone B, azonafide, BAY-43-9006, BAY80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, bilirubin dicitrate, BCX-1777, BKM-120, bleocin, BLP-25, BMS- 184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW2992 (afatinib, tomtobok), BIBF1120 (valgatef), BI836845, BI2536, BI6727, BI836845, BI847325, BI853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryocistatin-1, bortezomib, brostallicin, busulfan, BYL-719 , CA-4 prodrug, CA-4, CapCell, calcitriol, canertinib, canfosfamide, capecitabine, carboxyphthalatoplatin, CCl-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, cefratonin, ceftriaxone, celecoxib, celmoleukin, cemadotin, CH4987655 / RO-4987655, chlorotrianisene, cilengitide, cyclosporine, CDA-II, CDC-394, CKD-602, CKI-27, clofarabine, colchicine,Combretastatin A4, COT inhibitors, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibody, CP-461, CV-247, cyanomorpholinodoxorubicin, cytarabine, D24851, decitabine, deoxorubicin, deoxyrubicin, deoxycoformycin, depsipeptide, desoxyepothilone B, dexamethasone, dexrazoxanet, diethylstilbestrol, diflomotecan, didox, DM DC, dolastatin 10, doranidazole, DS-7423, E7010, E-6201, edatrexate, edotreotide, efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509, enzastaurin, enzalutamide, elsamitrucin, epothilone B, epratuzumab, ER-86526, erlotinib, ET-18-0CH3, ethinylcytidine, ethinyl estradiol, exatecan, exatecan mesylate, exemestane, exisulind, fenretinide, figitumumab, floxuridi Folic acid, FOLFOX, FOLFOX4, FOLFIRI, formestane, fotemustine, galarubicin, gallium maltolate, gefinitib, gemtuzumab, gimatecan, glufosfamide, GCS-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-112 0212 (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, idatrexate, idenestrol, IDN-5109,IGF-1R inhibitors, IMC-1C11, IMC-A12 (cixutumumab), Immunol, indisulam, interferon α-2a, interferon α-2b, pegylated interferon α-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib, ipilimumab, iproplatin, irofulven, isohomohalichondrin-B, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, Kahalid F, ketoconazole, KW -2170, KW-2450, lobaplatin, leflunomide, lenograstim, leuprolide, leuporelin, lexidronam, LGD-1550, linezolid, lutetium texaphyrin, lometrexol, losoxantrone, LU223651, lurtotecan, LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechloroethamine, MEK inhibitor, MEK-162, methyltestosterone, methylprednisolone, MEDI-573, MEN-10755, MDX-H210, MDX-44 7, 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, norelin, N-acetylcysteine, 6-benzylguanine, oblimersen, omeprazole, Oncophage, OncoVEX, GM-CSF, olmiplatin, ortataxel, OX4 4 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, 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, Piva, methylbutyrate, pixantrone, phenoxodiol O, PKI166, previtrexed, plicamycin, polyprenoic acid, porfiromycin, prednisone, prednisolone, quinamed, quinupristin, R115777, RAF-265, ramosetron, ranpirnase, RDEA-119 / BAY869766, RDEA-436, rebeccamycin analogs, receptor tyrosine kinase (RTK) inhibitors, revimid, RG-7167, RG-7304, RG -7421, RG-7321, RG7440, rhizoxin, rhu-MAb, linfavert, risedronate, rituximab, lobatumumab, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR109881A, rubidazone, rubitecan, R-flurbiprofen, RX-0201, S-9788, sabalubicin, SAHA, sargramostim, satraplatin, SB408075, Se-015 / Ve-015, SU5416, SU6 668, SDX-101, semustine, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine, suberanilohydroxamic acid, Sutent, T900607, T138067, TAK-733, TAS-103, tacedinaline, talaporfin, tarceva, tariquitar, tasisulam, taxotere, taxoplexin , tazarotene, tegafur, temozolamide, tesmilifene, testosterone, testosterone propionate, tesmilifene, tetraplatin, tetrodotoxin, tezacitabine, thalidomide, telarubicin, thymalfasin, thymectacin, tiazofurin, tipifarnib, tirapazamine, tocladesine, tomudex, tremofin, trabectedin, TransMID-107, transretinoic acidacid), traszutumab, tremelimumab, tretinoin, triacetyluridine, triapine, triciribine, trimetrexate, TLK-286TXD258, Tycarb / Tybarb, Urocidin, valrubicin, vatalanib, vincristine, vinflunine, virudin, WX-UK1, WX-554, Vectibix, Zero The compound is selected from the group consisting of tetracycline, ...

[0223] In one embodiment, the other therapeutic agent comprises a steroid, including dexamethasone, prednisolone, methylprednisolone, prednisone, hydrocortisone, triamcinolone, betamethasone, and cortivazol. In one embodiment, the other therapeutic agent comprises an antiemetic. Antiemetics include, but are not limited to, 5-HT3 receptor agonists (such as dolasetron, granisetron, ondansetron, tropisetron, palonosetron, and mirtazapine), dopamine agonists (such as domperidone, olanzapine, droperidol, haloperidol, chlorpromazine, prochlorperazine, alizapride, prochlorperazine, and metoclopramide), NK1 receptor antagonists (such as aprepitant and casopitant), and the like. ), antihistamines (such as cyclizine, diphenhydramine, dimenhydrinate, doxylamine, meclizine, promethazine, hydroxyzine), cannabinoids (such as cannabis, dronabinol, nabilone, and Sativex), benzodiazepines (such as midazolam and lorazepam), anticholinergics (such as hyoscine), trimethobenzamide, ginger, emetrol, propofol, peppermint, muscimol, and ajowan.

[0224] The pharmaceutical composition may be administered to a subject by any suitable route of administration. In one embodiment, the pharmaceutical composition is administered to a subject orally, parenterally, transdermally, or transmucosally. In one embodiment, the pharmaceutical composition is administered to a subject parenterally. In one embodiment, the pharmaceutical composition is administered to a subject by a parenteral route selected from the group consisting of intravenous (IV), subcutaneous (SC), and intramuscular (IM). In one embodiment, the pharmaceutical composition is administered to a subject by a route of administration selected from rectal and transdermal. In one embodiment, the pharmaceutical composition is administered to a subject in a dosage form selected from the group consisting of 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 the group consisting of a tablet, caplet, capsule, lozenge, syrup, solution, suspension, and elixir. In one embodiment, the pharmaceutical composition is administered to a subject in an oral dosage form selected from the group consisting of a tablet, hard shell capsule, soft gelatin capsule, beads, granules, agglomerates, powders, gels, solids, and semisolids.

[0225] In one embodiment, the pharmaceutical composition is administered to the subject in a dosage form selected from the group consisting of sustained release, controlled release, delayed release, and responsive release forms.

[0226] In one embodiment, the pharmaceutical composition is administered to the subject once daily. In one embodiment, the pharmaceutical composition is administered to the subject according to an infrequent dosing schedule (e.g., administered once a week or less). In one embodiment, the pharmaceutical composition is administered to the subject according to a frequent dosing schedule (e.g., administered twice a week or more). 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.

[0227] In one embodiment, a treatment method comprises administering to a subject in need of such treatment (i) a first therapeutic agent comprising a compound comprising an imipridone, such as 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; and further comprising assaying the expression of an endoplasmic reticulum (ER) stress response gene in a biological sample. In one embodiment, the ER stress response gene is selected from the group including, but not limited to, C / EBP-homologous protein (CHOP), activating transcription factor 3 (ATF3), and both CHOP and ATF3. In one embodiment, the ER stress response gene is selected from the group including, but not limited to, ATF3, activating transcription factor 4 (ATF4), CHOP, IRE1, binding immunoglobulin protein (BiP), eukaryotic translation initiation factor 2A (eIF2a), and X-box binding protein 1 (XBP1). The biological sample may be a tumor, peripheral blood mononuclear cell, or skin sample. The biological sample may be taken before, during, or after administration of the drug. In one embodiment, the method of treatment further comprises adjusting the dose 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 dose 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 an ER stress gene. In one embodiment, the method of treatment further comprises adjusting the dose 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 the ER stress gene.

[0228] In one embodiment, a treatment method comprises administering to a subject in need of such treatment (i) a first therapeutic agent comprising a compound comprising an imipridone, such as ONC201, 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; and further comprising assaying the expression of proteasome activity in a biological sample. In one embodiment, the proteasome activity can be chymotrysin-like, trypsin-like, and / or caspase-like activity. In one embodiment, the biological sample can be a tumor, peripheral blood mononuclear cells, or skin cells. The biological sample can be collected before, during, or after drug administration. In one embodiment, the method of treatment further comprises adjusting the dose to achieve 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% inhibition of proteasome activity. In one embodiment, the method of treatment further comprises adjusting the dose to achieve 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% inhibition of proteasome activity. In one embodiment, the treatment method further includes adjusting the dose to achieve 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% inhibition of proteasome activity.

[0229] In one aspect, the present invention provides a method of treatment comprising administering to a subject in need thereof a combination of a first therapeutic agent comprising an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt (e.g., a di- or tri-salt) thereof, and a second therapeutic agent, the method comprising: (i) administering to a subject a first therapeutic agent; (ii) waiting until a predetermined waiting period has elapsed after administration of the first therapeutic agent to the subject; and / or until the adverse event has resolved or is resolving; and (iii) administering to the subject a second therapeutic agent. wherein the predetermined waiting time is selected to provide a delayed therapeutic effect of the first therapeutic agent without increasing the risk of potential combined toxic effects of the first and second therapeutic agents. In one embodiment, the predetermined waiting time is determined based on the clearance rate of the first therapeutic agent compound or a metabolite thereof. In one embodiment, the predetermined waiting time is determined by a quantitative assessment of renal function and renal parameters. In one embodiment, the predetermined waiting time is determined by an assay that measures renal function, the assay being selected from the group consisting of: serum levels of the first therapeutic agent compound or a metabolite thereof; clearance rate of the first therapeutic agent compound or a metabolite thereof; and 24-hour urinary clearance of the first therapeutic agent compound or a metabolite thereof.

[0230] In one embodiment of the method of treatment, the predetermined waiting time is substantially equal to the time required for systemic clearance of the first therapeutic compound or its metabolites from the subject's body. In one embodiment of the method of treatment, the predetermined waiting time is substantially equal to the time required for renal clearance of the first therapeutic compound or its metabolites from the subject's body. In one embodiment of the method of treatment, the predetermined waiting time is substantially equal to the time required for hepatic clearance of the first therapeutic compound or its metabolites from the subject's body. In one embodiment of the method of treatment, the predetermined waiting time is substantially equal to the time required for total clearance of the first therapeutic compound or its metabolites from the subject's body. In one embodiment of the method of treatment, the predetermined waiting time is about 4 hours. In another embodiment, the waiting time is 1 day. In one embodiment, the waiting time is equal to the time required for total clearance of the first therapeutic compound or its metabolites from the subject's body. maxIn other embodiments, the waiting period is after the majority of adverse events have resolved or are in the process of being resolved. 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 within 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 above periods are considered the "treatment period."

[0231] If the order of administration is reversed, the timing of administration of the first therapeutic agent should be adjusted to coincide with the C of the second therapeutic agent (i.e., the drug administered first). max In one embodiment, administration of the first therapeutic agent can be after most or substantially all of the initially administered drug has been eliminated from the body or after the toxic effects of the initially administered drug have resolved or are being resolved.

[0232] In one embodiment, the treatment method further includes monitoring the level of the first therapeutic compound or a metabolite thereof in the subject using pharmacokinetic profiling. In some such embodiments, monitoring the level of the first therapeutic compound or a metabolite thereof in the subject using pharmacokinetic profiling includes generating a pharmacokinetic profile of the first therapeutic compound or a metabolite thereof in the subject using the concentrations of the first therapeutic compound or a metabolite thereof in at least two samples collected from the subject at time points suitable for generating the pharmacokinetic profile. In one embodiment that includes monitoring the level of the first therapeutic compound or a metabolite thereof in the subject using pharmacokinetic profiling, the sample is collected from the subject at the point of care or point of use via specimen collection or self-sample collection using a point-of-care or point-of-use device or on a matrix suitable for storing the sample before quantification in a laboratory. In one embodiment, the point-of-care or point-of-use device, respectively, is capable of quantifying the first therapeutic compound or a metabolite thereof. In one embodiment involving monitoring the level of a first therapeutic agent compound or its metabolite in a subject, one or more samples are collected from the subject at the point of care or point of use using a biopsy device and either analyzed by the point of care or point of use device or stored prior to laboratory analysis. In one embodiment, the biopsy samples are taken 3 to 8 hours apart after administering the first therapeutic agent to the subject. In one embodiment, the biopsy samples are taken 3 to 24 hours apart after administering the first therapeutic agent to the subject. In one embodiment, the biopsy samples are taken 8 to 24 hours apart after administering the first therapeutic agent to the subject. In one embodiment, the biopsy samples are taken 2 days apart after administering the first therapeutic agent to the subject. In one embodiment, the biopsy samples are taken 3 days apart after administering the first therapeutic agent to the subject. In one embodiment, the biopsy samples are taken 4 days apart after administering the first therapeutic agent to the subject. In one embodiment, the biopsy samples are taken 1 to 7 days apart after administering the first therapeutic agent.

[0233] In one embodiment, the pharmacokinetic profile comprises pharmacokinetic parameters suitable for guiding the administration of the first therapeutic agent to the subject being treated. In one embodiment of the method of treatment, the C of the first therapeutic agent after administration to the subject is max is in the range of about 1000 ng / dL to 1500 ng / dL during treatment. max is less than 1500 ng / dL and greater than 85 ng / dL during the treatment period. In one embodiment, the C after administering the first therapeutic agent to the subject max is in the range of about 1000 ng / mL to 1500 ng / mL during the treatment period. max is less than 1500 ng / mL and greater than 85 ng / mL during treatment.

[0234] In one embodiment, the maximum concentration ("C") of a first therapeutic agent in a subject's blood (whole blood, plasma, or serum) after administration of the therapeutic agent to the subject is measured. max ") are approximately 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 approximately 1500 ng / dL; approximately 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 C of 5, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 ng / dL to about 150 ng / dL; or a C of about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14, or 14.5 ng / dL to about 15 ng / dLmax is.

[0235] In one embodiment, the maximum concentration ("C") of the first therapeutic agent in the subject's blood (whole blood, plasma, or serum) following administration of the first therapeutic agent is measured. max ") are approximately 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 to about 1500 ng / mL; 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 C of about 5, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 ng / mL to about 150 ng / mL; or about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14, or 14.5 ng / mL to about 15 ng / mL max is.

[0236] In one embodiment, the maximum concentration ("C") of the first therapeutic agent in the subject's blood (whole blood, plasma, or serum) following administration of the first therapeutic agent is measured. max") is approximately 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 maximum concentration ("C") of the first therapeutic agent in the subject's blood (whole blood, plasma or serum) following administration of the first therapeutic agent is selected from the group consisting of: max ")C max is selected 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. max is selected from about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14 or 14.5 ng / dL.

[0237] In one embodiment, the C after administration of the first therapeutic agent maxare approximately 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, In one embodiment, the C after administration of the first therapeutic agent is selected from the group consisting of 1260, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, and 1490 ng / mL. max is selected 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 / mL. max is selected from about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14 or 14.5 ng / mL.

[0238] In one embodiment, the C after administration of the first therapeutic agent maxare approximately 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, 47 5, 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, 87 5, 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. maxare approximately 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 8, 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 9, 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. In one embodiment, the C after administration of the first therapeutic agent max is selected from 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.

[0239] In one embodiment, the C after administration of the first therapeutic agent maxare approximately 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, 47 5, 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, 87 5, 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 / mL. maxare approximately 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 8, 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 , 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 after administration of the first therapeutic agent max is selected from 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.

[0240] In one embodiment, the C of the first therapeutic agent after administration to the subject max In one embodiment, the C in the subject's blood (whole blood, plasma, or serum) after administration of the first therapeutic agent is in the range of about 85 ng / dL to 1500 ng / dL; about 8.5 ng / dL to 150 ng / dL; or about 0.85 ng / dL to 15 ng / dL. maxare approximately 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, 47 5, 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, 87 5, 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 approximately 1500 ng / dL;Approximately 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, 5 9, 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 6, 147, 148, or 149 ng / dL to about 150 ng / dL; or selected from 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;

[0241] In one embodiment, the C after administration of the first therapeutic agent max is in the range of about 85 ng / mL to 1500 ng / mL; about 8.5 ng / mL to 150 ng / mL; or about 0.85 ng / mL to 15 ng / mL. In one embodiment, the C after administration of the first therapeutic agent is in the range of about 85 ng / mL to 1500 ng / mL; about 8.5 ng / mL to 150 ng / mL; or about 0.85 maxare approximately 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, 47 5, 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, 87 5, 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;Approximately 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, 5 9, 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 6, 147, 148, or 149 ng / mL to about 150 ng / mL; or selected from 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;

[0242] In one embodiment, total drug exposure over time, measured as the area under the curve ("AUC") of a plot of the concentration of drug in a subject's blood (whole blood, plasma, or serum) after drug administration versus time after drug administration, is within the range of about 150 ng-hr / mL to about 8000 ng-hr / mL; about 15 ng-hr / mL to about 800 ng-hr / mL; or 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 800 ng-hr / mL and greater than or equal to 1.5 ng-hr / mL.

[0243] In one embodiment, the total drug exposure over time is an AUC of about 100 ng-hr / mL to about 8000 ng-hr / mL; about 10 ng-hr / mL to about 800 ng-hr / mL; or about 1 ng-hr / mL to about 80 ng-hr / mL. In one embodiment, the total drug exposure over time is an AUC of 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 an AUC of 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 an AUC of 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.

[0244] In one embodiment, the total drug exposure over time is an AUC of about 100 ng-hr / mL to about 8000 ng-hr / mL, about 10 ng-hr / mL to about 800 ng-hr / mL; or about 1 ng-hr / mL to about 80 ng-hr / mL. In one embodiment, the total drug exposure over time is an AUC of 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 an AUC of 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 an AUC of 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 an AUC of about 100 ng-hr / mL to about 200 ng-hr / mL; about 10 ng-hr / mL to about 20 ng-hr / mL; or about 1 ng-hr / mL to about 2 ng-hr / mL.

[0245] In one embodiment, the total drug exposure over time is an AUC selected from 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 nghr / mL. In one embodiment, the total drug exposure over time is an AUC selected from 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 nghr / mL. In one embodiment, the total drug exposure over time is an AUC selected from 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 nghr / mL.

[0246] In another aspect, the present invention provides a therapeutic method or use of a composition for treating a pathological condition, comprising administering to a subject in need thereof a combination of a first therapeutic agent and a second therapeutic agent, the method comprising: (i) administering to the subject a first therapeutic agent comprising an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof; (ii) monitoring the level of the first therapeutic compound or a metabolite thereof in the subject using pharmacokinetic profiling; and (iii) administering a second therapeutic agent in response to the level of the first therapeutic agent in the subject. In one embodiment, the monitoring step comprises generating a pharmacokinetic profile of the first therapeutic compound or its metabolite in the subject using concentrations of the first therapeutic compound or its metabolite in multiple samples collected from the subject at time points suitable for generating the pharmacokinetic profile. In one embodiment, at least two samples are collected at the point of care or point of use by specimen collection or self-sample collection with a point-of-care or point-of-use device or on a matrix suitable for storing the samples prior to quantification of the compound or its metabolite in a laboratory. In one embodiment, the point-of-care or point-of-use device is each capable of quantifying the compound or its metabolite. In one embodiment, the pharmacokinetic profile comprises pharmacokinetic parameters suitable for guiding administration of the compound or its salt 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 include AUC, AUC inf , T max , C max , time above threshold, steady-state concentration, absorption rate, clearance rate, distribution rate, terminal phase T-1 / 2, or a parameter derived from a compartmental pharmacokinetic (PK) analysis, including a non-compartmental PK analysis or a physiologically model-based compartmental PK analysis. In one embodiment, the method of treatment further comprises generating a report comprising the subject's pharmacokinetic profile. In one embodiment, the report comprises dosing recommendations based on the subject's pharmacokinetic profile. In one embodiment, a dose reduction of ONC201, its analog, or a pharmaceutically acceptable salt thereof is indicated to reduce the risk of toxicity based on the one or more pharmacokinetic parameters. In one embodiment, a dose reduction of the compound or its salt is indicated based on the time above threshold, and the threshold is the drug concentration or AUC above which toxicity occurs, AUC inf, mean residence time (MRT), an exponential function defining the pharmacokinetic profile, volume of distribution at steady state (Vss), volume of distribution at terminal phase (Vz), or a combination of pharmacokinetic variables that adequately describe the pharmacokinetic profile. In one embodiment, one or more pharmacokinetic parameters are used to direct dose adjustment of the compound or its salt to enhance efficacy. In one embodiment, AUC, AUC inf The dose of the compound or its salt is increased based on one or more of the following: MRT, an exponential function defining the pharmacokinetic profile, the volume of distribution at steady state (Vss), the volume of distribution at terminal phase (Vz), or a combination of pharmacokinetic variables that adequately describe the pharmacokinetic profile. In one embodiment, the dose of the compound or its salt is adjusted to within 5% to 25% of a desired target value. In one embodiment, each sample is applied to a point-of-care or point-of-use device for determining the concentration of the compound or its metabolites, the point-of-care or point-of-use device comprising a lateral flow strip having a structure and composition such that application of one or more samples to the lateral flow strip causes a fraction of the drug in the sample to bind to components of the lateral flow strip, thereby generating a detectable signal proportional to the concentration in the applied sample. In one embodiment, the sample is applied to a matrix suitable for storing the sample prior to quantification in the 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 LC-MS. In one embodiment, the pharmacokinetic parameters include at least one of steady state concentration, absorption, and terminal phase T1 / 2. In one embodiment, at least one of the samples is whole blood.

[0247] IX. Multidisciplinary therapy

[0248] In one aspect, the present invention provides a multimodality therapy in which the administration of an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof to a subject in need of treatment is supplemented with other therapies. In one embodiment, the multimodality therapy comprises administering to the subject a pharmaceutical composition comprising an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, in combination with radiation therapy or after determining that radiation has been ineffective. In one embodiment, the multimodality therapy comprises administering to the subject a pharmaceutical composition comprising an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, in combination with radiation therapy, wherein the administration of the pharmaceutical composition comprising an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof and radiation therapy are performed simultaneously or sequentially. In one embodiment, the multimodality therapy comprises administering to the subject a pharmaceutical composition comprising an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, in a sequential sequence in combination with radiation therapy. In one embodiment, the multimodality therapy comprises administering to a subject in need of such treatment a pharmaceutical composition comprising an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, simultaneously with radiation therapy. In one embodiment, the multimodality therapy is used to treat cancer. In one embodiment, the multimodality therapy comprises administering to a cancer subject in need of such treatment a pharmaceutical composition comprising an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, and irradiating the cancer cells with a radiation beam. In one embodiment, the multimodality therapy uses a conformal radiation therapy (CRT) technique to obtain a dose-volume histogram (DVH) to be prescribed to the cancer subject. In one embodiment, the multimodality therapy uses an intensity-modulated radiation therapy (IMRT) technique to irradiate the cancer cells. In one embodiment, the multimodality therapy uses a technique to compensate for the behavior of the subject's tumor during treatment (e.g., when radiation must be administered to a thoracic tumor that moves with the patient's breathing). For example, multimodality therapy uses four-dimensional computed tomography (4D CT) scanning technology to adjust the delivered radiation field to compensate for tumor behavior throughout the respiratory cycle.

[0249] Multimodality therapy can use any suitable type of radiation, including fractionated gamma radiation, IMRT (intensity-modulated radiation therapy), gamma knife, proton therapy, and brachytherapy. Radiation therapy and administration of imipridone, such as ONC201, its analogs, or pharmaceutically acceptable salts thereof, can be used to treat brain tumors, such as glioblastoma or brain metastases from lung cancer. Multimodality therapy 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, multimodality therapy includes using proton therapy to treat cancers, including brain tumors, prostate cancer, and tumors that are close to the organ and for which minimizing toxicity to nearby normal tissues is crucial.

[0250] In one embodiment, the multimodality therapy comprises administering to a cancer subject in need of such treatment a pharmaceutical composition comprising an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof, simultaneously or in combination with adoptive cell therapy (e.g., CAR-T (JCAR14, 15, 16, 17, KTE-C19, or CTL019); other T cells (AFM13); or NK (CDNO-109 or NK-92)).

[0251] In one embodiment, the multimodality therapy eliminates minimal residual disease without increasing the toxicity resulting from treatment with an imipridone, such as ONC201, an analog thereof, or a pharmaceutically acceptable salt thereof. In one embodiment, the multimodality therapy improves the prognosis of the subject being treated and / or reduces adverse side effects of the pathological state or condition.

[0252] X. Other Imipridone Derivatives, Analogues, and Salts

[0253] In one aspect, the present invention provides compounds and methods for making analogs of the compound of formula 10. Those skilled in the art will appreciate that the general principles and concepts described above in connection with ONC201 and compounds of formula 10 and salts thereof, including principles and concepts related to methods and pharmaceutical compositions, apply equally to the following analogs and salts thereof.

[0254] In one embodiment, the analog is a compound having a structure represented by the following structural formula (25):

[0255] [ka]

[0256] wherein R1, R2, R3, and R4 are, independently of one another, 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, R1, R2, R3, and R4 are optionally substituted. In one embodiment, some or all of the hydrogen atoms in R1, R2, R3, and R4 are replaced with deuterium. In another embodiment, R1, R2, R3, and R4 are not H, C, or C. 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone, C 1~4 Benzyl-piperazine and C 1~4 alkylthienyl, independently selected from the group consisting of C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone and C 1~4 Benzyl-piperazine is optional, C1~4 In another embodiment, R, R, R, and R are independently selected from the group consisting of H, CH, CHPh, CH-((2-Cl)-Ph), CH-(2-thienyl), CHCHPh, CHCH(4-N-benzyl-piperazine), CH-(2,4-diF-Ph), CH-((2-CH)-Ph), CHCHOHPh, and (CH)CO-4F-Phare.

[0257] In one embodiment, the analog is a compound having a structure represented by the following structural formula (26):

[0258] [ka]

[0259] wherein R1 and R2 independently 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, R1 and R2 are selected from the group consisting of H, C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone, C 1~4 Benzyl-piperazine and C 1~4 alkylthienyl, independently selected from the group consisting of C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone and C 1~4 Benzyl-piperazine is optional, C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4In one embodiment, R is selected from the group consisting of H, CH, CHPh, CH-((2-Cl)-Ph), CH-(2-thienyl), CHCHPh, CHCH(4-N-benzyl-piperazine), CH-(2,4-diF-Ph), CH-((2-CH)-Ph), CHCHOHPh, and (CH)CO-4F-Ph. In one embodiment, R2 is 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.

[0260] In one embodiment, R1 optionally contains one or more of the following substituents, alone or in combination, at the ortho, meta and / or para positions 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 X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R2 is one or more of the following substituents, alone or in combination, at the ortho, meta, and / or para positions 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 of 2 to 20, and X is a halogen.

[0261] In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 It is substituted with alkyl or halo.

[0262] In one embodiment, R2 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 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, R2 is a substituted or unsubstituted heterocycloalkylalkyl, such as a morpholinoalkyl or piperazinylalkyl group. In one embodiment, R2 is a substituted or unsubstituted heteroarylalkyl, such as an isoxazolidinylmethyl or pyridylmethyl group. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, —CH 3 , —CF 3 , and —OCH 3 .

[0263] In one embodiment, the analog is a compound having a structure represented by the following structural formula (27):

[0264] [ka]

[0265] wherein R1 is 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, R1 is H, C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone, C 1~4 Benzyl-piperazine and C 1~4 alkylthienyl, C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone and C 1~4 Benzyl-piperazine is optional, C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, R is selected from the group consisting of H, CH, CHPh, CH-((2-Cl)-Ph), CH-(2-thienyl), CHCHPh, CHCH(4-N-benzyl-piperazine), CH-(2,4-diF-Ph), CH-((2-CH)-Ph), CHCHOHPh, and (CH)CO-4F-Ph.

[0266] In one embodiment, R1 optionally contains one or more of the following substituents, alone or in combination, at the ortho, meta and / or para positions 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+1wherein 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. In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is a C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 It is substituted with alkyl or halo.

[0267] In one embodiment, the analog is a compound having a structure represented by the following structural formula (28):

[0268] [ka]

[0269] wherein R1 and R2 independently 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, R1 and R2 are selected from the group consisting of H, C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone, C 1~4 Benzyl-piperazine and C 1~4 alkylthienyl, independently selected from the group consisting of C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone and C 1~4 Benzyl-piperazine is optional, C 1~4 Alkyl, C 1~4Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, R is selected from the group consisting of H, CH, CHPh, CH-((2-Cl)-Ph), CH-(2-thienyl), CHCHPh, CH-(2,4-diF-Ph), CH-((2-CH)-Ph), CHCHOHPh, CHCH(4-N-benzyl-piperazine), and (CH)CO-4F-Ph. In one embodiment, R2 is 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, when R1 is CH2Ph, R2 is not CH2-(2-CH3-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2-CH3-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2,4-diF-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(4-CF3-Ph).

[0270] In one embodiment, R1 optionally contains one or more of the following substituents, alone or in combination, at the ortho, meta and / or para positions 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 X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R2 is one or more of the following substituents, alone or in combination, at the ortho, meta, and / or para positions of the benzyl ring: -CH3, -NO2, -OCH3, -CXH2, -CX2H, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X2p+1 , -OCX3 or -OC p X 2p+1 wherein p is an integer of 2 to 20, and X is a halogen.

[0271] In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 It is substituted with alkyl or halo.

[0272] In one embodiment, R2 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 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, R2 is a substituted or unsubstituted heterocycloalkylalkyl, such as a morpholinoalkyl or piperazinylalkyl group. In one embodiment, R2 is a substituted or unsubstituted heteroarylalkyl, such as an isoxazolidinylmethyl or pyridylmethyl group. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, —CH 3 , —CF 3 , and —OCH 3 .

[0273] In one embodiment, the analog is a compound having a structure represented by the following structural formula (29):

[0274] [ka]

[0275] wherein R1 and R2 independently 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, R1 and R2 are selected from the group consisting of H, C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone, C 1~4 Benzyl-piperazine and C 1~4 alkylthienyl, independently selected from the group consisting of C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone and C 1~4 Benzyl-piperazine is optional, C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4In one embodiment, R is selected from the group consisting of H, CH, CHPh, CH-((2-Cl)-Ph), CH-(2-thienyl), CHCHPh, CHCH(4-N-benzyl-piperazine), CH-(2,4-diF-Ph), CH-((2-CH)-Ph), CHCHOHPh, and (CH)CO-4F-Ph. In one embodiment, R2 is 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, when R1 is CH2Ph, R2 is not CH2-(2-CH3-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2-CH3-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2,4-diF-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(4-CF3-Ph).

[0276] In one embodiment, R1 optionally contains one or more of the following substituents, alone or in combination, at the ortho, meta and / or para positions 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 X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R2 is one or more of the following substituents, alone or in combination, at the ortho, meta, and / or para positions 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 pX 2p+1 wherein p is an integer of 2 to 20, and X is a halogen.

[0277] In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 It is substituted with alkyl or halo.

[0278] In one embodiment, R2 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 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, R2 is a substituted or unsubstituted heterocycloalkylalkyl, such as a morpholinoalkyl or piperazinylalkyl group. In one embodiment, R2 is a substituted or unsubstituted heteroarylalkyl, such as an isoxazolidinylmethyl or pyridylmethyl group. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, —CH 3 , —CF 3 , and —OCH 3 .

[0279] In one embodiment, the analog is a compound having a structure represented by the following structural formula (30):

[0280] [ka]

[0281] wherein R1 and R2 independently 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, R1 and R2 are selected from the group consisting of H, C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone, C 1~4 Benzyl-piperazine and C 1~4 alkylthienyl, independently selected from the group consisting of C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone and C 1~4 Benzyl-piperazine is optional, C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4In one embodiment, R is selected from the group consisting of H, CH, CHPh, CH-((2-Cl)-Ph), CH-(2-thienyl), CHCHPh, CHCH(4-N-benzyl-piperazine), CH-(2,4-diF-Ph), CH-((2-CH)-Ph), CHCHOHPh, and (CH)CO-4F-Ph. In one embodiment, R2 is 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, when R1 is CH2Ph, R2 is not CH2-(2-CH3-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2-CH3-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2,4-diF-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(4-CF3-Ph).

[0282] In one embodiment, R1 optionally contains one or more of the following substituents, alone or in combination, at the ortho, meta and / or para positions 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 X is a halogen, including F, Cl, Br, or I, preferably F, Cl, or Br, more preferably F or Cl. In one embodiment, R2 is one or more of the following substituents, alone or in combination, at the ortho, meta, and / or para positions 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 pX 2p+1 wherein p is an integer of 2 to 20, and X is a halogen.

[0283] In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 It is substituted with alkyl or halo.

[0284] In one embodiment, R2 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 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, R2 is a substituted or unsubstituted heterocycloalkylalkyl, such as a morpholinoalkyl or piperazinylalkyl group. In one embodiment, R2 is a substituted or unsubstituted heteroarylalkyl, such as an isoxazolidinylmethyl or pyridylmethyl group. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, —CH 3 , —CF 3 , and —OCH 3 .

[0285] In one embodiment, the analog is a compound having a structure represented by the following structural formula (31):

[0286] [ka]

[0287] wherein R1 and R2 independently 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, R1 and R2 are selected from the group consisting of H, C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone, C 1~4 Benzyl-piperazine and C 1~4 alkylthienyl, independently selected from the group consisting of C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4 Alkyl phenyl ketone and C 1~4 Benzyl-piperazine is optional, C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4In one embodiment, R is selected from the group consisting of H, CH, CHPh, CH-((2-Cl)-Ph), CH-(2-thienyl), CHCHPh, CHCH(4-N-benzyl-piperazine), CH-(2,4-diF-Ph), CH-((2-CH)-Ph), CHCHOHPh, and (CH)CO-4F-Ph. In one embodiment, R2 is 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, when R1 is CH2Ph, R2 is not CH2-(2-CH3-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2-CH3-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2,4-diF-Ph). In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(4-CF3-Ph).

[0288] In one embodiment, R1 optionally contains one or more of the following substituents, alone or in combination, at the ortho, meta and / or para positions 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 X is a halogen, including F, Cl, Br, or I; preferably F, Cl, or Br; more preferably F or Cl. In one embodiment, R2 is one or more of the following substituents, alone or in combination, at the ortho, meta, and / or para positions 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 pX 2p+1 wherein p is an integer of 2 to 20, and X is a halogen.

[0289] In one embodiment, R1 is H. In one embodiment, R1 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 It is substituted with alkyl or halo.

[0290] In one embodiment, R2 is a substituted or unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In one embodiment, arylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 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, R2 is a substituted or unsubstituted heterocycloalkylalkyl, such as a morpholinoalkyl or piperazinylalkyl group. In one embodiment, R2 is a substituted or unsubstituted heteroarylalkyl, such as an isoxazolidinylmethyl or pyridylmethyl group. In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, —CH 3 , —CF 3 , and —OCH 3 .

[0291] In one embodiment, the present invention provides a compound represented by the following structural formula (29):

[0292] [ka]

[0293] wherein R1 and R2 are independently selected from H, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, and acyl radicals. In one embodiment, R1 is CH2Ph ​​and R2 is CH2-(2-CH3-Ph), which is the linear isomer of ONC201 (i.e., TIC-10) that lacks anticancer activity (Jacob et al., Angew. Chem. Int. Ed., (2014) 53:6628; Wagner et al., Oncotarget (2015) 5(24):12728).

[0294] [ka]

[0295] 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).

[0296] In one embodiment, R1 and R2 are H, 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 Alkylpyridinyl, C 1~4 Alkylisoxazolidinyl, C 1~4 Alkylmorpholinyl, C 1~4 Alkylthiazolyl and C 1~4 alkylpyrazinyl, independently selected from the group consisting of C 1~4 Alkyl, C 1~4 Alkylphenyl, C 1~4Alkyl phenyl ketone, C 1~4 Benzyl-piperazine, C 1~4 Alkylthienyl, C 1~4 Alkylpyridinyl, C 1~4 Alkylisoxazolidinyl, C 1~4 Alkylmorpholinyl, C 1~4 Alkylthiazolyl and C 1~4 Alkylpyrazinyl is optionally C 1~4 Alkyl, C 1~4 Alkoxyl, hydroxyl, perhalogenated C 1~4 In one embodiment, R and / or R are substituted with alkyl or halo. In one embodiment, R and / or R are substituted or unsubstituted aryl alkyl or heteroaryl alkyl. In one embodiment, heteroaryl alkyl is C 1~4 Alkylpyrrolyl, C 1~4 Alkylfuryl, C 1~4 Alkylpyridyl, C 1~4 Alkyl-1,2,4-thiadiazolyl, C 1~4 Alkylpyrimidyl, C 1~4 Alkylthienyl, C 1~4 Alkylisothiazolyl, C 1~4 Alkyl imidazolyl, C 1~4 Alkyltetrazolyl, C 1~4 Alkylpyrazinyl, C 1~4 Alkylpyrimidyl, C 1~4 Alkylquinolyl, C 1~4 Alkylisoquinolyl, C 1~4 Alkylthiophenyl, C 1~4 Alkylbenzothienyl, C 1~4 Alkylisobenzofuryl, C 1~4 Alkylpyrazolyl, C 1~4 Alkyl indolyl, C 1~4 Alkylpurinyl, C 1~4 Alkylcarbozolyl, C 1~4 Alkylbenzimidazolyl and C 1~4 alkylisoxazolyl.

[0297] In one embodiment, R1 and / or R2 optionally have one or more of the following substituents on the benzyl ring: 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 , S.R. m , N.R. m R n , N.R. m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m benzyl substituted with R m and R n are independently selected from H or C1-C4 alkyl; 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.

[0298] XI. Working Examples

[0299] It should be understood that the descriptions and examples set forth below are merely intended to be illustrative and are not intended to limit the scope of the present disclosure. The examples below are intended to illustrate the disclosed embodiments and should not be construed as being limited thereto. In addition to those described below, other compounds can be prepared according to the following reaction schemes or suitable variations or modifications thereof.

[0300] Example 1. Synthesis of 2-chlorobenzylamino-2-imidazoline hydroiodide

[0301] To a stirred solution of 2-methylthio-2-imidazoline hydroiodide (244 mg, 1.00 mMol) in dry 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 min. The solution was cooled to room temperature, filtered through a sintered funnel, washed with cold dioxane (2 mL), and dried under vacuum. Compound 4·HI (R2 = 2-chlorobenzyl) (242 mg, 72%) was obtained as a white solid and used without further purification.

[0302] Example 2. Synthesis of 2-chlorobenzylamino-2-imidazoline

[0303] To a stirred solution of 2-chlorobenzylamino-2-imidazoline hydroiodide (242 mg, 0.72 mMol) in water (3 mL) at 7 °C was added 1.0 N sodium hydroxide (2 mL). The reaction mixture was stirred under argon at 7 °C for 30 min. After that, methylene chloride (5 mL) was added and the mixture was stirred for an additional 5 min. The reaction mixture was extracted with methylene chloride (2 × 2.5 mL). The organic layer was dried over anhydrous NaSO, filtered, and evaporated. The resulting free base (150 mg, 100%) was obtained as a viscous liquid and was used in the next reaction without further purification. MS (ESI) 210 (M+H).

[0304] Example 3. Synthesis of methyl-1-benzyl 4-oxo-3-piperidinecarboxylate (compound (6))

[0305] To stirred methyl-1-benzyl 4-oxo-3-piperidinecarboxylate hydrochloride (5.7 g, 20 mMol) in ethyl acetate (50 mL) was added triethylamine (6 mL) at 7 °C. The reaction mixture was stirred at 7 °C for 30 minutes under an argon atmosphere. The reaction mixture was extracted with ethyl acetate (2 × 50 mL) and washed with water (50 mL). The organic layer was dried over anhydrous NaSO, filtered, and evaporated. The free base residue (5, R = benzyl) obtained as a viscous oil was used in the next reaction without further purification. MS (ESI) 248 (M + H).

[0306] Example 4. Synthesis of ONC202 (compound (14))

[0307] To a solution of 2-chlorobenzylamino-2-imidazoline (150 mg, 0.72 mMol) and methyl 1-benzyl 4-oxo-3-piperidinecarboxylate (5, R = benzyl) (195 mg, 0.79 mMol) in 1-butanol (2 mL) was added PPTS (10 mg), and the mixture was stirred at room temperature for 48 h. The reaction mixture was then refluxed at 125-130 °C for 2 h. The solvent was removed in vacuo, extracted with ethyl acetate (10 mL), and washed with saturated sodium bicarbonate solution (2 × 10 mL) and water (10 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and evaporated. The crude free base was purified by RP HPLC (10%-40% acetonitrile / water) to give ONC902 TFA salt as a white solid (228 mg, 50% yield). MS (ESI) 407 (M+H).

[0308] Using the same procedure starting from various benzylamines, various analogs such as ONC203, 204, 205, 206, 912, 210, 211, 212, 213, 214, 217, 218, 219, 220, 221, 222, 223, 224, 225 and 226 were prepared.

[0309] Example 5. Synthesis of ONC207 (compound (19))

[0310] Dimethyl carbonate (4.32 g, 48.0 mMol) was added dropwise to a suspension of 60% sodium hydride (3.5 g, 88 mMol) in dry toluene (50 mL) under nitrogen at room temperature over 0.5 h. After adding a few drops of methanol, 1-tert-butoxycarbonyl-4-piperidone (4.8 g, 24 mMol) dissolved in dry toluene (20 mL) was added dropwise to the reaction mixture over 1 h with stirring at 80 °C. The reaction mixture was stirred at the same temperature for 3 h, then cooled to 0 °C (ice bath) and adjusted to pH 6-6.5 with acetic acid. The resulting cooled 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 under vacuum to give methyl-1-tertbutoxycarbonyl-4-oxo-3-piperidinecarboxylate (5.0 g, 80%), which was used in the next reaction without further purification.

[0311] To 2-methylbenzylamino-2-imidazoline (190 mg, 1 mMol), methyl 1-tert-butoxycarbonyl-4-oxo-3-piperidinecarboxylate (315 mg, 1.1 mMol) in 1-butanol (2 mL) was added PPTS (10.0 mg), and the mixture was stirred at room temperature for 48 h. The reaction mixture was then refluxed at 125-130 °C for 2 h. The solvent was removed in vacuo, extracted with ethyl acetate (10 mL), and 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 and purified by RP HPLC (10%-40% acetonitrile / water) to give ONC907 (262 mg, 50%) as a TFA salt as a white solid. MS(ESI) 297(M+H).

[0312] Example 6. Synthesis of ONC209 (compound (21))

[0313] A mixture of ONC907 (100 mg, 0.2 mMol), phenylethyl 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 h. The solvent was removed in vacuo, extracted with ethyl acetate (10 mL), and washed with water (5 mL). The organic layer was dried over anhydrous NaSO, filtered, and evaporated. The crude free base was purified by RP HPLC (10%-40% acetonitrile / water) to give ONC209 (62 mg, 50%) as the TFA salt as a white solid. MS (ESI) 401 (M+H).

[0314] The same procedure starting from various halides was used to give ONCs 215 and 214. Starting from various benzylamines and using the analogous procedure of Examples 1 and 5, compounds 227, 228, 229, 230, 231, 232, 233, 234, 235, and 236 were prepared. The intermediate compounds where R1 is H were then treated with various halides as above.

[0315] Compound ONC216 was prepared from ONC215 by treatment with TFA.

[0316] The precursor NH compound prepared in the same manner as in Example 5 was reacted and treated with styrene oxide to prepare compound (72).

[0317] Example 7. Synthesis of ONC208 (compound (20))

[0318] To a solution of 2-methylbenzylamino-2-imidazoline (190.0 mg, 1.0 mMol) and methyl 1-methyl 4-oxo-3-piperidinecarboxylate (185.0 mg, 1.0 mMol) in 1-butanol (2.0 mL) was added PPTS (10.0 mg), and the mixture was stirred at room temperature for 48 h. The reaction mixture was then refluxed at 125-130 °C for 2 h. The solvent was removed in vacuo, extracted with ethyl acetate (10 mL), and 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 using 10%-40% acetonitrile and water to give ONC208 (270.0 mg, 50%) as a TFA salt as a white solid. MS (ESI) 311 (M+H).

[0319] Example 8. Synthesis of ONC201 (compound (1))

[0320] Compound (3) (239.7 g, 0.845 mol, 1.6 equiv.) was added in small portions to 800 mL of stirred saturated NaHCO3 in a 2 L round-bottom flask. n-Butanol (500 mL) was added to the resulting mixture, which was stirred for 30 min and then transferred to a separatory funnel. The organic phase containing compound (4) was separated and transferred to a 2 L three-necked round-bottom flask equipped with mechanical stirring, a N2 inlet, a thermocouple, a condenser, and a Dean-Stark trap. Compound (5) (100 g, 0.528 mol, 1 equiv.) and pyridinium p-toluenesulfonate (PPTS) (6.63 g, 0.026 mol, 5 mol%) were added to the contents of the flask. The resulting mixture was heated to reflux for 6 h. Water in the reaction mixture was separated into the Dean-Stark trap as needed. The reflux temperature was increased from 93 °C to 118 °C. T...

Claims

1. A drug for treating central nervous system cancer having an epigenetically suppressed unmethylated O(6)-methylguanine-DNA methyltransferase (MGMT) gene, the drug comprising a compound of the following formula (I) or a pharmaceutically acceptable salt thereof: 【Chemistry 1】

2. The method of claim 1, wherein the central nervous system cancer is selected from the group consisting of meningioma, ependymoma, glioma, neuroblastoma, and diffuse intrinsic pontine glioma.

3. The drug according to claim 1 or 2, for use in humans.

4. The drug according to any one of claims 1 to 3, wherein the pharmaceutically acceptable salt is a dihydrochloride salt.

5. The agent according to any one of claims 1 to 4, wherein the central nervous system cancer is highly sensitive to modulation by one or more of dopamine receptors (DRs) and G protein-coupled receptors (GPCRs).

Images