Hexahydroimidazopyridopyrimidone, its analogs and salts, and therapeutic uses thereof

Abstract

The present application relates to hexahydroimidazopyridopyrimidone, its analogs and salts, and therapeutic uses thereof. In particular, the present application relates to a compound of formula (10), and a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same, and pharmaceutical uses thereof.

Description

[0001] The present application is a divisional application of Chinese patent application No. 201680008476.7 filed on January 29, 2016, with the invention name of “7-benzyl-4-(2-methylbenzyl)-2,4,6,7,8,9-hexahydroimidazo[1,2-A]pyrido[3,4-E]pyrimidin-5(1H)-one, its analogs, and its salts, and methods for their use in treatment”. Technical Field

[0002] The present application relates to a compound of formula (10) or a pharmaceutically acceptable salt thereof, and its pharmaceutical use. Background Art

[0003] TNF-related apoptosis-inducing ligand (TRAIL; Apo2L) is an endogenous protein that selectively induces apoptosis in cancer cells. TRAIL is a potent inducer of apoptosis in a wide range of human cancer cell lines - via the pro-apoptotic death receptor 4 (DR4; TRAIL-R1) and death receptor 5 (DR5; TRAIL-R2) on the cell surface, by engaging either the extrinsic or intrinsic apoptotic pathways. TRAIL plays a direct role in tumor suppression during immune surveillance, but this anti-tumor mechanism is lost during disease progression. The ability of TRAIL to selectively induce apoptosis in cancer cells has led to ongoing clinical trials of recombinant TRAIL, as well as longer-lived TRAIL agonist antibodies targeting either of its two pro-apoptotic death receptors.

[0004] Although it is effective, recombinant TRAIL has properties that limit effectiveness, such as short serum half-life, stability, cost, and delivery. Delivery of recombinant TRAIL or TRAIL agonist antibodies to the brain is limited due to the inability of recombinant TRAIL and TRAIL agonist antibodies to cross the blood-brain barrier. Therefore, there is a continuing need for anticancer compositions and methods. Summary of the invention

[0005] In one aspect, provided herein is a compound of formula (10): wherein R1 and R2 independently represent hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, thioaralkyl and acyl radicals, wherein when R1 represents CH2Ph, R2 does not represent CH2-((2-CH3)-Ph). In some embodiments, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine, C 1-4 Alkylthienyl, C1-4 Alkylpyridyl, C 1-4 Alkylisoxazolidinyl, C 1-4 Alkylmorpholinyl, C 1-4 Alkylthiazolyl and C 1-4 Alkylpyrazinyl, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine, C 1-4 Alkylthienyl, C 1-4 Alkylpyridyl, C 1-4 Alkylisoxazolidinyl, C 1-4 Alkylmorpholinyl, C 1-4 Alkylthiazolyl, and C 1-4 The alkylpyrazinyl group is optionally substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In some embodiments, R1 and / or R2 are substituted or unsubstituted aralkyl or heteroaralkyl. In some embodiments, heteroaralkyl is selected from C 1-4 Alkylpyrrolyl, C 1-4 Alkyl furanyl, C 1-4 Alkylpyridyl, C 1-4 Alkyl-1,2,4-thiadiazolyl, C 1-4 Alkylpyrimidinyl, C 1-4 Alkylthienyl, C 1-4 Alkylisothiazolyl, C 1-4 Alkyl imidazoline, C 1-4 Alkyl tetrazolyl, C 1-4 Alkylpyrazinyl, C 1-4 Alkylpyrimidinyl, C 1-4 Alkylquinolyl, C 1-4 Alkylisoquinolyl, C 1-4 Thioalkylphenyl, C 1-4 Alkylbenzothiophene, C 1-4 Alkyl isobenzofuranyl, C 1-4 Alkylpyrazolyl, C 1-4 Alkyl indolyl, C 1-4 Alkyl purine, C 1-4 Alkylcarbazolyl, C 1-4 Alkylbenzimidazolinyl and C 1-4In some embodiments, R1 and / or R2 are substituted or unsubstituted benzyl or phenethyl. In some embodiments, R1 and / or R2 are benzyl optionally substituted on the benzyl ring with one or more of the following substituents: X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 ,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ;

[0006] R m and R n Independently selected from hydrogen or C1-C4 alkyl;

[0007] wherein p is an integer from 2 to 20 and X represents a halogen.

[0008] In one embodiment, compound (10) has compound (90) The structure of, wherein R2 is as defined above, and wherein R b1 , R b2 , R b3 , R b4 and R b5 Each is independently selected from hydrogen, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, -OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2Rm , C(O)R m and C(O)OR m ;

[0009] R m and R n Independently selected from hydrogen or C1-C4 alkyl;

[0010] wherein p is an integer from 2 to 20 and X represents a halogen.

[0011] In one embodiment, compound (10) has compound (40) wherein R1 is as defined above, and wherein R a1 , R a2 , R a3 , R a4 and R a5 are each independently selected from hydrogen, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 ,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ;

[0012] R m and R n Independently selected from hydrogen or C1-C4 alkyl;

[0013] wherein p is an integer from 2 to 20 and X represents a halogen.

[0014] In one embodiment, compound (10) has compound (50): The structure wherein R1 is as defined above, and wherein R b is selected from X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ; and where R a1 , R a2 , R a4 and R a5 are each independently selected from hydrogen, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 , -OCX3, -OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ;

[0015] R m and R n Independently selected from hydrogen or C1-C4 alkyl;

[0016] wherein p is an integer from 2 to 20 and X represents a halogen.

[0017] In one embodiment, compound (10) has compound (80) The structure of R a1 , R a2 , R a3 , R a4 , R as , R b1 , R b2 , R b3 , R b4 and R b5are each independently selected from hydrogen, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 ,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ;

[0018] R m and R n Independently selected from hydrogen or C1-C4 alkyl;

[0019] wherein p is an integer from 2 to 20 and X represents a halogen.

[0020] On the other hand, the present invention provides a pharmaceutical composition comprising a compound of formula (10) or a pharmaceutically acceptable salt thereof. In one embodiment, the pharmaceutical composition comprises a pharmaceutically acceptable salt of a compound of formula (10). In one embodiment, the salt is a pharmaceutically acceptable monosalt of a compound of formula (10). In one embodiment, the salt is a pharmaceutically acceptable disalt of a compound of formula (10). In one embodiment, the pharmaceutically acceptable salt is selected from hydrochloride, hydrobromide, bisulfate, sulfate, phosphate, fumarate, succinate, oxalate and lactate, bisulfate, hydroxy, tartrate, nitrate, citrate, bitartrate, carbonate, malate, maleate, fumarate, sulfonate, methanesulfonate, formate, acetate and carboxylate. In one embodiment, the pharmaceutically acceptable salt is selected from p-toluenesulfonate, benzenesulfonate, methanesulfonate, oxalate, succinate, tartrate, citrate, fumarate, glucuronate, ascorbate and maleate. In one embodiment, the pharmaceutically acceptable salt is selected from ammonium, sodium, potassium, calcium, magnesium, zinc, lithium and / or other counterions, such as methylamino, dimethylamino, diethylamino and triethylamino counterions. In one embodiment, the pharmaceutical composition comprises the hydrochloride disalt or hydrobromide disalt of the compound of formula (10). In some embodiments, the pharmaceutical composition of the present invention comprises a pharmaceutically acceptable carrier.

[0021] In some embodiments, the pharmaceutical composition of the present invention comprises a second therapeutic agent. In one embodiment, the second therapeutic agent is an anticancer agent. In one embodiment, the anticancer agent is a mitotic inhibitor. In one embodiment, the anticancer agent is selected from: paclitaxel, docetaxel and combinations thereof. In an alternate embodiment, the second therapeutic agent is an anti-angiogenic agent. In one embodiment, the anti-angiogenic agent is bevacizumab. In one embodiment, the second therapeutic agent is administered as part of a combination therapy to treat the patient. In one embodiment, the details of the combination therapy are included in the package insert of the compound of formula (10).

[0022] In some embodiments, the pharmaceutical composition is formulated for oral administration.

[0023] In another aspect, the present invention provides a method of treatment. In one embodiment, the method of treatment comprises administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula (10) or a pharmaceutically acceptable salt thereof.

[0024] In one embodiment, the method of treatment comprises administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula (10) or a pharmaceutically acceptable salt thereof. In one embodiment, the method of treatment comprises administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula (10) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0025] In some embodiments, the method of treatment further comprises administering an additional therapeutic agent. In one embodiment, the additional therapeutic agent comprises an anticancer agent. In one embodiment, the additional anticancer agent comprises an antimitotic agent. In one embodiment, the additional anticancer agent comprises paclitaxel, docetaxel, bevacizumab, or a combination thereof.

[0026] In one embodiment, the method of treatment further comprises testing a sample obtained from the subject undergoing treatment for tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL). In one embodiment, the sample is a blood sample.

[0027] In one embodiment of the method of treatment, the subject undergoing treatment has cancer or is at risk of cancer. In one embodiment, the cancer is selected from colon cancer, breast cancer, glioblastoma multiforme, endometrial cell lymphoma, and colorectal cancer. In one embodiment, the cancer is selected from actinic keratosis, Barrett's esophagus, atrophic gastritis, dyskeratosis congenita, subarachnoid dysphagia, lichen planus, oral submucous fibrosis, solar elastosis, cervical dysplasia, leukoplakia, and erythema.

[0028] In one embodiment of the method of treatment, the pharmaceutical composition is administered by an oral administration route. In one embodiment, the pharmaceutical composition is administered by an administration route selected from the group consisting of intravenous, rectal, nasal, pulmonary, epidural, ophthalmic, aural, intra-arterial, topical, intracardiac, intracerebroventricular, intradermal, intramuscular, intraperitoneal, intraosseous, intrathecal, intravesical, subcutaneous, transdermal, transmucosal, sublingual, oral, vaginal, and inhalation administration routes.

[0029] In one embodiment, the present invention provides a method for treating a subject having brain cancer or at risk of brain cancer, the method comprising: administering to the subject a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula (10) or a pharmaceutically acceptable salt thereof. In one embodiment, the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

[0030] In one embodiment, the present invention provides a method of treatment, comprising administering to a subject a pharmaceutical composition comprising: a pharmaceutically effective amount of a compound of formula (10) or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

[0031] In another aspect, the present invention provides a method of treating a subject in need of such treatment, the method comprising:

[0032] (i) administering to a subject a first therapeutic agent comprising a compound of formula (10) or a pharmaceutically acceptable salt thereof;

[0033] (ii) after the time at which the first therapeutic agent is administered to the subject, waiting until a predetermined waiting time has elapsed; and

[0034] (iii) administering a second therapeutic agent to the subject, wherein the predetermined waiting period is selected to obtain a delayed therapeutic effect of the first therapeutic agent without increasing the risk of an increase in, or a reduced risk of, a possible combined toxic effect of the first and second therapeutic agents.

[0035] In another aspect, the present invention provides a method of treating a subject in need of such treatment, the method comprising:

[0036] (i) administering to a subject a first therapeutic agent comprising a compound of formula (10) or a pharmaceutically acceptable salt thereof;

[0037] (ii) monitoring the level of the compound of formula (10) or a salt thereof, or a metabolite thereof in a subject using a pharmacokinetic assay; and

[0038] (iii) administering a second therapeutic agent conditioned on the level of the first therapeutic agent in the subject.

[0039] In another aspect, the present invention provides a method comprising:

[0040] (i) administering to the subject a first therapeutic agent comprising a compound of formula (10) or a pharmaceutically acceptable salt thereof; and

[0041] (ii) administering the second therapeutic agent under conditions consistent with the expected half-life of the compound of formula (10) in the subject undergoing treatment.

[0042] In another aspect, provided herein are analogs of compounds of formula (10): Formula (1): and their pharmaceutically acceptable salts, and their preparation methods. In one aspect, a method of treatment is provided herein. In one embodiment, the method of treatment administers a pharmaceutical composition to a subject, the composition comprising: a pharmaceutically effective amount of a compound of formula (10) or formula (1) or its analog, or a pharmaceutically acceptable salt thereof. In one embodiment, the pharmaceutical composition comprises a pharmaceutically acceptable carrier. In one embodiment, the subject undergoing treatment has cancer or is at risk of cancer. In one embodiment, the subject is a human.

[0043] In another aspect, the present invention provides a method of treating a subject in need of such treatment, the method comprising:

[0044] (i) administering to the subject a first therapeutic agent, including an analog of compound (1) (eg, a compound of formula (10)) or a pharmaceutically acceptable salt thereof; and

[0045] (ii) administering the second therapeutic agent in accordance with the condition that the adverse event of the first therapeutic agent has been resolved or is being resolved. In some embodiments, the adverse event of the first therapeutic agent is related to the blood level of the agent or metabolite in the subject undergoing treatment.

[0046] In another aspect, the present invention provides a kit for monitoring compound (1) (e.g., compound of formula (10)) analogs, or pharmaceutically acceptable salts thereof, or metabolites thereof in individuals treated with compound (1) or analogs thereof, or pharmaceutically acceptable salts thereof, or metabolites thereof using pharmacokinetic analysis, the kit comprising a plurality of point-of-care devices or point-of-use devices capable of quantifying the drug in at least two samples or matrices suitable for storing those samples prior to quantification in a laboratory. In some embodiments, the kit further comprises instructions for collecting and / or storing at least two samples.

[0047] In some embodiments, the method of treatment administers a pharmaceutical composition to a subject, the pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula (1) or formula (10) or an analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) monitoring the level of the compound or its pharmaceutically acceptable salt, or its metabolite in the subject using pharmacokinetic analysis, wherein the dose of the compound or its salt is selected to maintain a concentration of the compound or its metabolite of at least about 400 ng / mL in the subject's whole blood, plasma, serum, or cerebrospinal fluid within 4 hours after treatment. In some embodiments, the subject undergoing treatment has cancer or a risk of cancer. In some embodiments, the method of treatment further comprises a step of waiting for a predetermined waiting time between one or more repetitions of step (i) (e.g., the waiting time is the same as the length of the first time interval). In some embodiments, the subject is a human.

[0048] In some embodiments, the method of treatment comprises administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula (1) or (10), or an analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) monitoring the level of the compound or its metabolite in the subject using a pharmacokinetic assay, wherein the dose of the compound or its salt is selected to achieve an AUC of at least about 3,500 hr-ng / mL for the compound or its metabolite in the whole blood, plasma, serum or cerebrospinal fluid of the subject over a period of 2, 6, 12, 24, 48, 72 or more hours, or extrapolated to infinity.

[0049] In some embodiments, the treatment method comprises administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula (1) or formula (10) or an analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) monitoring the level of compound (1), or a pharmaceutically acceptable salt thereof, or a metabolite thereof in the subject using a pharmacokinetic analysis, wherein the dose of the compound or its salt is selected to result in an undetectable concentration of the compound or its metabolite in the whole blood, plasma, serum or cerebrospinal fluid of the subject at 3 days, 4 days, 5 days, 6 days or 7 days after treatment.

[0050] In some embodiments, the method of treatment comprises: (i) administering multiple doses of a therapeutic agent, including a compound of formula (1) or formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof, within a first time interval (e.g., 7 days), wherein multiple doses are administered during a second time interval within the first time interval (e.g., a first time interval of two to five consecutive days for a week); and (ii) repeating step (i) one or more times. In some embodiments, the subject undergoing treatment has cancer or is at risk of cancer. In some embodiments, the method of treatment further comprises a step of waiting for a predetermined waiting time (e.g., the waiting time is the same length as the first time interval) between one or more repetitions of step (i). In some embodiments, the subject is a human.

[0051] In some embodiments, the pharmaceutical composition is administered to the subject once a day. In some embodiments, the pharmaceutical composition is administered to the subject according to an infrequent dosing regimen (e.g., administered once a week or less frequently). In some embodiments, the pharmaceutical composition is administered to the subject according to a frequent dosing regimen (e.g., administered more than once a week). In some embodiments, the pharmaceutical composition is administered to the subject once a week. In some embodiments, the pharmaceutical composition is administered to the subject once every four weeks. In some embodiments, The pharmaceutical composition is administered to the subject twice a week. In some embodiments, the pharmaceutical composition is administered to the subject three times a week. In some embodiments, the pharmaceutical composition is administered to the subject four times a week. In some embodiments, the pharmaceutical composition is administered to the subject once every two weeks. In some embodiments, the pharmaceutical composition is administered to the subject once every three weeks. In some embodiments, the pharmaceutical composition is administered to the subject once a week, once every two weeks, once every three weeks, once every four weeks, or a combination thereof in a repetitive cycle.

[0052] In one aspect, the present invention provides a method for identifying whether a subject with a disease may respond to a treatment regimen. In some embodiments, the method includes (i) obtaining a biological sample from the subject; (ii) measuring the RNA modification level and / or expression or activity level of one or more proteins in Tables 1 and 3 in the sample, and / or expressing the presence or absence of one or more mutations in at least one gene of the protein of one or more Tables 1 and 3; (iii) comparing the measured level and / or mutations found in the sample with those of a predetermined standard; and (iv) based on the level of the mutation measured and / or found in the sample and those of a predetermined standard, determining whether the subject may respond to a treatment regimen. In one aspect, the present invention provides a method for providing a prognosis for a subject with a disease. In some embodiments, the method comprises: (i) obtaining a biological sample from the subject; (ii) measuring the RNA modification level and / or expression or activity level of one or more proteins in Tables 1 and 3 in the sample, and / or the presence or absence of one or more mutations in at least one gene expressing one or more proteins in Tables 1 and 3; (iii) comparing the measured levels and / or mutations found in the sample with those of predetermined standards; and (iv) determining the prognosis of the subject based on the levels of mutations measured and / or found in the sample with those of predetermined standards.

[0053] In some embodiments, the subject has cancer or is at risk of cancer. In some embodiments, the treatment regimen comprises administering an effective amount of compound (1) or an analog thereof. In one embodiment, the RNA is mRNA. In one embodiment, the RNA is miRNA. In one embodiment, the RNA modification is methylation. In one embodiment, the RNA modification is N 6 -methyladenosine (m 6 A). In one embodiment, the RNA modification is m 6 In one embodiment, the subject is a human. In one embodiment, the subject is a domesticated pet, such as a cat or dog. In one embodiment, the protein is an RNA reader, such as YTHDF3. In one embodiment, the protein is an RNA writer. In one embodiment, the protein is an RNA wiper.

[0054] In one aspect, provided herein are methods for identifying and treating subjects with a condition and a possible response to a treatment regimen described herein. In some embodiments, the method comprises (i) identifying whether the subject has a condition that may respond to a treatment regimen described herein; and (ii) using a treatment regimen to treat subjects determined to be likely to respond to the treatment regimen. In some embodiments, the subject has cancer or a risk of cancer. In some embodiments, the treatment regimen comprises administering an effective amount of compound (1) or its analog.

[0055] In one aspect, a method for evaluating monitoring a subject with a disease and experiencing the effect of treatment according to a treatment regimen described herein is provided herein. In some embodiments, the method includes (i) obtaining a biological sample from the subject; (ii) measuring the RNA modification level and / or expression or activity level of one or more proteins in Tables 1 and 3 in the sample, and / or the presence or absence of one or more mutations in at least one gene expressing one or more proteins in Tables 1 and 3; (iii) comparing the measured levels and / or mutations found in the sample with those of a predetermined standard; and (iv) determining whether the subject responds to the treatment regimen based on the level of the mutation measured and / or found in the sample with those of a predetermined standard.

[0056] In some embodiments, the subject has cancer or is at risk of cancer. In some embodiments, the treatment regimen comprises administering an effective amount of compound (1) or an analog thereof. In one embodiment, the RNA is mRNA. In one embodiment, the RNA is miRNA. In one embodiment, the RNA modification is methylation. In one embodiment, the RNA modification is N 6 -methyladenosine (m 6 A). In one embodiment, the RNA modification is m 6 In one embodiment, the subject is a human. In one embodiment, the subject is a domesticated pet, such as a cat or dog. In one embodiment, the protein is an RNA reader, such as YTHDF3. In one embodiment, the protein is an RNA writer. In one embodiment, the protein is an RNA wiper.

[0057] In one aspect, provided herein is a method for screening potential treatments for a disease. In some embodiments, the method comprises (i) contacting a protein of Tables 1 and 3 with a test compound suspected of being a treatment for the disease; (ii) measuring the binding affinity or interaction of the test compound and the protein; and (iii) comparing the binding affinity or interaction of the test compound and a predetermined threshold, wherein the affinity or interaction of the test compound is comparable to or greater than the threshold and indicates treatment of the disease. In some embodiments, the disease is cancer. In some embodiments, the predetermined threshold is the affinity or interaction of compound (1) or its analog and the protein. In some embodiments, the predetermined threshold is the affinity or interaction of compound (1) and the protein. In some embodiments, the protein is an RNA reader, such as YTHDF3. In some embodiments, the protein is an RNA writer. In some embodiments, the protein is an RNA wiper.

[0058] In one aspect, provided herein is a method for screening potential treatments for a disease. In some embodiments, the method includes (i) contacting a protein of Tables 1 and 3 with a reference compound in a disease, wherein the reference compound and the protein bind or interact; (ii) contacting a protein of Tables 1 and 3 with a test compound suspected of being a treatment for the disease; (iii) measuring the binding affinity or interaction of the reference and test compounds and the protein; and (iv) comparing the binding affinity or interaction of the reference and test compounds, wherein the comparable or stronger affinity or interaction of the test compound relative to the reference compound indicates the treatment of the disease. In one embodiment, the reference compound is compound (1) or an analog thereof. In one embodiment, the reference compound is compound (1). In one embodiment, the disease is cancer. In one embodiment, the protein is an RNA reader, such as YTHDF3. In one embodiment, the protein is an RNA writer. In one embodiment, the protein is an RNA wiper.

[0059] In one aspect, provided herein is a method for screening for potential treatment of a disease. In some embodiments, the method includes (i) if any, using a processor to model the combination or interaction of a 3-dimensional structure of a test compound suspected of treating a disease and a 3-dimensional structure of a protein of Tables 1 and 3; (ii) using a processor, determining the binding affinity or interaction of the test compound structure and the protein structure; and (iii) using a processor, comparing the binding affinity or interaction of the test compound with a predetermined threshold, wherein the binding affinity or interaction of the test compound is comparable or greater than the threshold, indicating cancer treatment. In one embodiment, the predetermined threshold is the affinity or interaction of compound (1) or its analog and the protein. In one embodiment, the predetermined threshold is the affinity or interaction of compound (1) and the protein. In some embodiments, the disease is cancer. In one embodiment, the protein is an RNA reader, such as YTHDF3. In one embodiment, the protein is an RNA writer. In one embodiment, the protein is an RNA wiper.

[0060] In one aspect, a method for screening potential treatments for a disease is provided herein. In some embodiments, the method includes (i) using a processor to model a combination or interaction of a 3-dimensional structure of a reference compound and a 3-dimensional structure of a protein of Tables 1 and 3; (ii) if any, using a processor to model a combination or interaction of a 3-dimensional structure of a test compound suspected of treating a disease and a 3-dimensional structure of a protein; (iii) using a processor to determine the binding affinity or interaction of the reference and test compounds with the protein; and (iv) using a processor to compare the binding affinity or interaction of the reference and test compound structures, wherein the comparable or stronger affinity or interaction of the test compound relative to the reference compound indicates the treatment of the disease. In one embodiment, the reference compound structure is the structure of compound (1) or an analog thereof. In some embodiments, the reference compound structure is the structure of compound (1). In some embodiments, the disease is cancer. In some embodiments, the protein is an RNA reader, such as YTHDF3. In some embodiments, the protein is an RNA writer. In some embodiments, the protein is an RNA wiper.

[0061] In one aspect, provided herein is a method for treating a subject with a disorder by administering an agent that modulates the transcription, translation or biological activity of YTHDF3 or other proteins in Tables 1 and 3 or nucleic acids encoding these proteins. Agents include, but are not limited to, siRNA, antisense nucleic acids, ribozymes, triple helix forming agents, antibodies and polypeptides, and small molecule compounds. Preferably, siRNA, antisense nucleic acids, ribozymes, triple helix forming agents inhibit the translation or transcription of one or more genes in Tables 1 and 3. To achieve this, the oligonucleotides used are designed based on the unique associated sequences of the target gene. Various gene therapy methods can be used according to the present invention to modulate the expression of the genes in Tables 1 and 3. For example, antisense DNA molecules can be engineered and used to block the translation of YTHDF3 mRNA in vivo. Alternatively, ribozyme molecules may be designed to cut and destroy YTHDF3 mRNA in vivo. Alternatively, oligonucleotides designed to hybridize with the 5′ region of the YTHDF3 gene (including the region upstream of the coding sequence) and form a triple helix structure block or reduce the transcription of the YTHDF3 gene. In another alternative, nucleic acid encoding a full-length wild-type YTHDF3 message can be introduced in vivo into cells that would otherwise fail to produce sufficient amounts or no wild-type YTHDF3 gene product.

[0062] In one embodiment, the pharmaceutical composition comprises a pharmaceutically acceptable carrier. In some embodiments, the subject has cancer or is at risk of cancer. In some embodiments, the subject is infected with a virus or is at risk of being infected with a virus. In some embodiments, the subject is a human. In some embodiments, the dose of compound (1) or a pharmaceutically acceptable salt thereof is in the range of about 125 mg to about 625 mg. In some embodiments, a sample obtained from the subject is tested for cracked and / or total cytokeratin-18 to determine whether the treatment regimen should continue based on the results of the cracked and / or total cytokeratin-18 test. In some embodiments, the treatment regimen described herein also includes the step of administering a second therapeutic agent, wherein compound (1) or a pharmaceutically acceptable salt thereof, or an analog thereof, is administered before, simultaneously with, or after the second therapeutic agent.

[0063] In one aspect, a method for identifying whether a subject with a condition is likely to respond to a treatment regimen described herein is provided herein. In some embodiments, the method comprises (i) obtaining a biological sample from the subject; (ii) measuring the expression, post-translational modification, or activity level, or mutation of eIF2-α, ATF4, CHOP or DR5 in the sample; (iii) comparing the level of mutation measured and / or found in the sample with those of a predetermined standard; and (iv) determining whether the subject is likely to respond to a treatment regimen based on the level of mutation measured and / or found in the sample with those of a predetermined standard. In another aspect, a method for evaluating the effect of a treatment regimen described herein, monitoring or providing a prognosis for a subject with a condition is provided herein. In some embodiments, the method comprises (i) obtaining a biological sample from the subject; (ii) measuring the expression, post-translational modification, or activity level, or mutation of eIF2-α, ATF4, CHOP or DR5 in the sample; (iii) comparing the level of mutation measured and / or found in the sample with those of a predetermined standard; and (iv) determining a prognosis or determining whether the subject responds to a treatment regimen based on the level of mutation measured and / or found in the sample with those of a predetermined standard. In some embodiments, the subject has cancer or is at risk of cancer. In some embodiments, the subject has cancer or is at risk of cancer. In some embodiments, the subject is infected with a virus or is at risk of being infected with a virus. In some embodiments, the treatment regimen comprises administering an effective amount of compound (1) or a pharmaceutically acceptable salt thereof, or an analog thereof.

[0064] In one aspect, the present invention provides a method for identifying whether a subject with a disease may respond to a treatment regimen described herein. In some embodiments, the method includes (i) obtaining a biological sample from the subject; (ii) measuring the expression, post-translational modification, or activity level, or mutation of at least one dopamine receptor in the sample; (iii) comparing the level of mutations measured and / or found in the sample with those of a predetermined standard; and (iv) based on the level of mutations measured and / or found in the sample with those of a predetermined standard, determining whether the subject may respond to the treatment regimen. On the other hand, the present invention provides a method for evaluating the effect of the treatment regimen described herein, monitoring, or providing a prognosis for a subject with a disease. In some embodiments, the method includes (i) obtaining a biological sample from the subject; (ii) measuring the expression, post-translational modification, or activity level, or mutation of at least one dopamine receptor in the sample; (iii) comparing the level of mutations measured and / or found in the sample with those of a predetermined standard; and (iv) based on the level of mutations measured and / or found in the sample with those of a predetermined standard, determining the prognosis or determining whether the subject responds to the treatment regimen. In some embodiments, the subject has cancer or has a risk of cancer. In some embodiments, the subject has cancer or is at risk of cancer. In some embodiments, the treatment regimen comprises administering an effective amount of compound (1) or a pharmaceutically acceptable salt thereof, or an analog thereof. In some embodiments, the dopamine receptor is selected from DRD2, DRD2S, DRD2L and DRD3. In some embodiments, the dopamine receptor is from the D2-like family of dopamine receptors.

[0065] In one aspect, the present invention provides a method for screening potential treatments for a disease. In some embodiments, the method includes: (i) contacting at least one G protein-coupled receptor (GPCR) with a test compound suspected of being a treatment for the disease; (ii) measuring the binding affinity, interaction or GPCR signal of the test compound and the GPCR; and (iii) comparing the binding affinity or interaction of the test compound and a predetermined threshold, wherein the GPCR antagonism or GPCR signal antagonism of the test compound is comparable or greater than the threshold, indicating cancer treatment. In one embodiment, the disease is cancer. In one embodiment, the predetermined threshold is the GPCR antagonism or GPCR signal antagonism of compound (1) or a pharmaceutically acceptable salt thereof, or an analog thereof.

[0066] In one aspect, the present invention provides a method for screening potential treatments for a disease. In some embodiments, the method includes (i) contacting at least one dopamine receptor with a test compound suspected of being a treatment for the disease; (ii) measuring the binding affinity or interaction of the test compound and at least one dopamine receptor; and (iii) comparing the binding affinity or interaction of the test compound and a predetermined threshold, wherein the inhibition of at least one dopamine receptor by the test compound is comparable to or greater than the threshold, indicating cancer treatment. In some embodiments, the disease is cancer. In some embodiments, the dopamine receptor is selected from DRD2, DRD2S, DRD2L and DRD3. In some embodiments, the inhibition of the D2-like family of dopamine receptors indicates the treatment of the disease. In some embodiments, the inhibition of DRD2 and DRD3 dopamine receptors indicates the treatment of the disease. In some embodiments, the predetermined threshold is the inhibition of at least one dopamine receptor by compound (1) or a pharmaceutically acceptable salt thereof, or an analog thereof and a dopamine receptor.

[0067] In one aspect, the present invention provides a method for screening potential treatments for a disease. In some embodiments, the method includes (i) if there is, using a processor to model the combination or interaction of a 3-dimensional structure of a test compound suspected of treating the disease and a 3-dimensional structure of at least one dopamine receptor; (ii) using a processor, determining the binding affinity or interaction of the test compound structure and at least one dopamine receptor structure; and (iii) using a processor, comparing the binding affinity or interaction of the test compound with a predetermined threshold, wherein the inhibition of at least one dopamine receptor by the test compound is comparable to or greater than the threshold, indicating cancer treatment. In some embodiments, the disease is cancer. In some embodiments, the dopamine receptor is selected from DRD2, DRD2S, DRD2L and DRD3. In some embodiments, the inhibition of the D2-like family of dopamine receptors indicates the treatment of the disease. In some embodiments, the inhibition of DRD2 and DRD3 dopamine receptors indicates the treatment of the disease. In some embodiments, the predetermined threshold is the inhibition of at least one dopamine receptor by compound (1) or a pharmaceutically acceptable salt thereof, or an analog thereof and a dopamine receptor.

[0068] In one aspect, provided herein are methods for treating and evaluating the effect of treatment in a subject with a condition. In some embodiments, the method comprises (i) treating the subject according to the treatment methods described herein; (ii) evaluating the effect of treatment as described herein. In some embodiments, the subject has cancer or is at risk of cancer. In one embodiment, the treatment regimen comprises administering an effective amount of compound (1) or a pharmaceutically acceptable salt thereof or an analog thereof.

[0069] The foregoing summary, as well as the following detailed description of embodiments of compositions and methods of treatment, will be better understood when read in conjunction with the appended claims.It is to be understood, however, that the invention is not limited to the precise arrangements and instrumentalities described herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0070] 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 of exemplary embodiments. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities shown.

[0071] In the attached picture:

[0072] Figure 1 Dose-response relationships are shown, showing the effects of various concentrations of compound (1) on the viability of tumor and normal cells;

[0073] Figure 2 The cell viability test in human fetal lung fibroblast (MRC-5) cells after 72 hours of treatment with compound (1) is illustrated.

[0074] Figure 3 Antagonism of ONC201 at dopamine receptors (DRD1, DRD2S, DRD2L, DRD3, DRD4, and DRD5) is shown.

[0075] Figure 4 Soluble prolactin detected by ELISA testing in peripheral blood of tumor patients at baseline and in real-time before and after a single ONC201 dose (PO 125-625 mg) is shown. Sampling time points post-treatment included 6 hours, 1, 2, 7 days, and 21 days post-treatment.

[0076] Figure 5A and Figure 5B Shown is the relationship between the mean ONC201 plasma concentration and time after the first dose of ONC201. Figure 5A Plasma concentrations are shown as the mean for each dose cohort. Figure 5B Plasma concentrations for subjects treated with 625 mg are shown in . Error bars represent standard deviation.

[0077] Fig. 6A and Figure 6B The AUC and C of ONC201 are shown. max Individual measurements relative to dose. *625 mg dose group based on expected dose proportionality line for males.

[0078] Fig. 7A and Figure 7B The M30 test rates relative to time after a single dose for the four patients and relative to tumor type are shown separately.

[0079] Figure 8 The in vitro sensitivity of ONC201 to various tumor types as determined in a large panel of cell lines from the Genomic Drug Sensitivity of Cancer Regime (GDSC) is shown. DETAILED DESCRIPTION

[0080] Scientific terms used herein are intended to have the meanings commonly understood by one of ordinary skill in the art. These terms are found defined and used illustratively in various standard references, including J. Sambrook and DW Russell, 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; B. Alberts et al., Molecular Biology of the Cell, 4th Ed., Garland, 2002; DL Nelson and MM Cox, Lehninger Principles of Biochemistry, 4th Ed., VH Freeman & 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. Vintersten, R. Behringer, Manipulating the Mouse Embryo: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory Press; Dec. 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 foregoing references are incorporated herein by reference in their entirety.

[0081] The singular terms "a," "an," and "the" are not limiting and include plural referents unless expressly specified or the context clearly dictates otherwise.

[0082] The term "substituted", if used herein, means that any one or more hydrogen atoms on the designated atom are replaced with the indicated group of choice, provided that the normal valence of the designated atom is not exceeded and the substitution results in a stable compound. When the substituent is keto (i.e., =O), 2 hydrogens on the atom are replaced. Ketoaromatic substituents are not present on aromatic moieties. Ring double bonds are double bonds formed between two adjacent ring atoms (e.g., C=C, C=N, or N=N).

[0083] When any variable (e.g., R4) occurs more than once in any constituent or type of compound, its definition at each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group is shown as substituted with 0-3 R4 moieties, the group may be optionally substituted with up to three R4 moieties, and R4 at each occurrence is independently selected from the definition of R4. In addition, combinations of substituents and / or variables are permissible, but only if such combinations result in stable compounds.

[0084] When an atom or chemical moiety is followed by a subscripted numerical range (e.g., C 1-6 ), you will realize that this means that all numbers in that range and all intermediate ranges are included. For example, "C 1-6 "Alkyl" is meant to include alkyl groups having 1, 2, 3, 4, 5, 6, 6-1, 1-5, 1-4, 1-3, 1-2, 2-6, 2-5, 2-4, 2-3, 3-6, 3-5, 3-4, 4-6, 4-5 and 5-6 carbons.

[0085] As used herein, "alkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. 1-6Alkyl is intended to include C1, C2, C3, C4, C5 and C6 alkyl groups. Examples of alkyl include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl sec-butyl, tert-butyl, n-pentyl, sec-pentyl, neopentyl and n-hexyl. In certain embodiments, straight or branched alkyl has six or less carbon atoms (e.g., straight C1-C6, branched C3-C6) in its main chain, and in other embodiments, straight or branched alkyl has four or less carbon atoms. Similarly, cycloalkyl has three to eight carbon atoms in its ring structure, and in other embodiments, cycloalkyl has five or six carbon atoms in the ring structure. Most preferably (C1-C6) alkyl, particularly ethyl, methyl, isopropyl, isobutyl, n-pentyl, n-hexyl and cyclopropylmethyl.

[0086] As used herein, the term "substituted alkyl" refers to an alkyl group as defined above, substituted by one, two or three substituents selected from halogen, -OH, alkoxy, -NH2, -N(CH3)2, -C(=O)OH, trifluoromethyl, -C≡N, -C(=O)O(C1-C4)alkyl, -C(=O)NH2, -SO2NH2, -C(=NH)NH2, -NO2, preferably containing one or two substituents selected from halogen, -OH, alkoxy, -NH2, trifluoromethyl, -N(CH3)2 and -C(=O)OH, more preferably selected from halogen, alkoxy and -OH. Examples of substituted alkyl groups include, but are not limited to, 2,2-difluoropropyl, 2-carboxycyclopentyl and 3-chloropropyl.

[0087] Unless otherwise specified, "lower alkyl" includes alkyl groups as defined above but having 1 to 6 carbon atoms in their backbone structure, preferably 1 to 4. "Lower alkenyl" and "lower alkynyl" have chain lengths of 2-6 carbon atoms, preferably 2-4 carbon atoms.

[0088] "Alkenyl" includes similar unsaturated aliphatic groups and possible replacement of the above-mentioned alkyl in length, but contains at least one double bond. For example, the term "alkenyl" includes straight chain alkenyl (e.g., vinyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl), branched alkenyl, cycloalkenyl (e.g., alicyclic) (e.g., cyclopropenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or alkenyl substituted cycloalkenyl, and cycloalkyl or cycloalkenyl substituted alkenyl. In certain embodiments, straight or branched alkenyl has six or less carbon atoms (e.g., C2-C6 for straight chain, C3-C6 for branched chain) in its main chain. Similarly, cycloalkenyl can have three to eight carbon atoms in its ring structure, and in some embodiments, cycloalkenyl has five or six carbons in the ring structure. The term "C2-C6" includes alkenyl containing 2-6 carbon atoms. The term "C3-C6" includes alkenyl groups containing 3 to 6 carbon atoms.

[0089] "Alkynyl" includes unsaturated aliphatic groups of similar length and possible replacement of the above-mentioned alkyl, but contains at least one triple bond. For example, "alkynyl" includes straight chain alkynyl (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl), branched chain alkynyl and cycloalkyl or cycloalkenyl substituted alkynyl. In certain embodiments, straight chain or branched chain alkynyl has 6 or less carbon atoms (e.g., straight chain C2-C6, branched C3-C6) on its main chain. The term "C2-C6" includes alkynyl containing 2-6 carbon atoms. The term "C3-C6" includes alkynyl containing 3 to 6 carbon atoms.

[0090] As used herein, "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic group in which each atom (i.e., backbone atom) forming the ring is a carbon atom. In one embodiment, the cycloalkyl group is saturated or partially unsaturated. In another embodiment, the cycloalkyl group is fused to an aromatic ring. The cycloalkyl group includes a group having 3 to 10 ring atoms. Illustrative examples of cycloalkyl groups include, but are not limited to, the following parts:

[0091]

[0092] Monocyclic cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Bicyclic cycloalkyl includes, but is not limited to, tetrahydronaphthyl, indanyl and tetrahydropentene. Polycyclic cycloalkyl includes adamantane and norbornane. The term cycloalkyl includes "unsaturated non-aromatic carbocyclic group" or "non-aromatic unsaturated carbocyclic group", which all refer to non-aromatic carbocyclic rings as defined herein, which contain at least one carbon-carbon double bond or one carbon-carbon triple bond.

[0093] As used herein, "cycloalkylalkyl" refers to an alkyl group substituted with a cycloalkyl group. Example cycloalkylalkyl groups include cyclopropylalkyl, cyclohexylalkyl, and the like.

[0094] As used herein, "heterocycloalkyl" refers to a non-aromatic heterocycle, wherein one or more ring atoms can be a heteroatom, such as O, N or S atoms. Heterocycloalkyl groups can include monocyclic or polycyclic (e.g., with 2,3 or 4 fused rings) ring systems and spirocycles. Example heterocycloalkyl groups include morpholino, thiomorpholino, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, 2,3-dihydrobenzofuranyl, 1,3-benzodioxolane, benzo-1,4-dioxane, piperidinyl, pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, imidazolyl, etc. Also included in the definition of heterocycloalkyl is a portion of an aromatic ring having one or more aromatic rings fused to a non-aromatic heterocycle (i.e., having the same key therewith), such as phthalimido, naphthyl imino and heterocyclic benzo derivatives. Heterocycloalkyl groups with one or more fused aromatic rings are connected by aromatic or non-aromatic moieties. Also included in the definition of heterocycloalkyl may be one or more of the ring atoms may be substituted with 1 or 2 oxygen or sulfur groups. In some embodiments, heterocycloalkyl groups have 1 to about 20 carbon atoms, and in other embodiments, from about 3 to about 20 carbon atoms. In some embodiments, heterocycloalkyl groups contain or In some embodiments, the heterocycloalkyl group has or In some embodiments, the heterocycloalkyl group contains In some embodiments, the heterocycloalkyl group contains Three keys.

[0095] As used herein, "heterocycloalkylalkyl" refers to an alkyl group substituted by a heteroalkyl group. Example heterocycloalkylalkyl groups include morpholinoalkyl and piperazinylalkyl, among others.

[0096] As used herein, "aryl" refers to monocyclic or polycyclic (eg, having 2, 3, or 4 fused rings) aromatic hydrocarbons, such as phenyl, naphthyl, anthracenyl, phenanthrenyl, and the like. In some embodiments, aryl has carbon atoms.

[0097] As used herein, "aralkyl" refers to an alkyl group substituted by an aryl group. Example aralkyl groups include benzyl and phenethyl.

[0098] As used herein, "heteroaryl" refers to an aromatic heterocycle having at least one heteroatom ring member such as sulfur, oxygen or nitrogen. Heteroaryl groups include monocyclic and polycyclic (e.g., having 2, 3 or 4 fused rings) systems. Any annular N atom in the heteroaryl group may also be oxidized to form an N-oxygen moiety. Examples of heteroaryl groups include, but are not limited to, pyridyl, N-oxypyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazinyl, furanyl, quinolyl, isoquinolyl, thienyl, imidazolinyl, thiazolyl, indolyl, pyrrolyl, oxazolyl, benzofuranyl, benzothienyl, benzothiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thienadiazolyl, isothiazolyl, benzothienyl, purinyl, carbazolyl, benzimidazolinyl, indolinyl, etc. In some embodiments, the heteroaryl group has 1 to about 20 carbon atoms, and in other embodiments, from about 3 to about 20 carbon atoms. In some embodiments, the heteroaryl group contains or In some embodiments, the heteroaryl group has or Heteroatoms.

[0099] As used herein, "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl group. An example of a heteroaryl group is pyridinylmethyl.

[0100] As used herein, "halogen" or "halogen" refers to a fluorine, chlorine, bromine or iodine atom, preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine. The term "halogen" refers to a moiety in which all hydrogens are replaced by halogen atoms.

[0101] The term "haloalkyl" refers to an alkyl moiety having a halogen atom replacing a hydrogen atom on one or more carbons of the hydrocarbon backbone. C1-C6 haloalkyl is intended to include a straight or branched chain alkyl group having six or less carbon atoms in its backbone, and a halogen atom replacing a hydrogen atom on one or more carbons of the hydrocarbon backbone.

[0102] The term "alkoxy" or "alkoxy" includes substituted and unsubstituted alkyl, alkenyl and alkynyl groups covalently linked to an oxygen atom. C1-C6 alkoxy refers to a six moiety with a small number of carbon atoms in the hydrocarbon backbone. Examples of alkoxy (or alkoxy groups) include methoxy, ethoxy, isopropoxy, propoxy, butoxy and pentoxy. Preferred are (C1-C3) alkoxy, particularly ethoxy and methoxy. Examples of substituted alkoxy include halogen alkoxy.

[0103] The term "hydroxy" or "hydroxyl" includes groups having either -OH or -O-.

[0104] The present invention also includes pharmaceutically acceptable salts of the chemical substances described herein. As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds, whose parent compounds are modified by converting existing acid or base moieties into their salt forms. Pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali metal or organic salts of acidic residues such as carboxyl acids, etc. The pharmaceutically acceptable salts of the present invention include conventional non-toxic salts of the parent compound formed, for example from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present invention are synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, these salts can be prepared by reacting the free acid or base forms of these compounds with stoichiometric amounts of an appropriate base or acid in water or in an organic solvent or in a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17 th ed., Mack Publishing Company, Easton, Pa., 1985, p.1418, Journal of Pharmaceutical Science, 66, 2 (1977), and PH. Stahl and CG Wermuth, editors, Handbook of Pharmaceutical Salts: Properties, Selection and Use, 2 nd Revised edition, Weinheim / Zürich: Wiley-VCH / VHCA (2011), the entire contents of which are incorporated herein by reference.

[0105] Examples of suitable inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid or hydrobromic acid, while examples of suitable organic acids may include carboxylic acid, sulfonic acid or sulfonic acid, such as acetic acid, tartaric acid, lactic acid, propionic acid, glycolic acid, malonic acid, maleic acid, fumaric acid, tannic acid, succinic acid, alginic acid, benzovaleric acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, cinnamic acid, mandelic acid, citric acid, maleic acid, salicylic acid, trifluoroacetic acid, 3-aminosalicylic acid, ascorbic acid, aconitic acid, nicotinic acid, isonicotinic acid, oxalic acid, gluconic acid, amino acids, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid or naphthalene-2-sulfonic acid. Examples of suitable inorganic bases may include sodium hydroxide, potassium hydroxide and ammonia, and examples of suitable organic bases are amines, for example tertiary amines such as trimethylamine, triethylamine, pyridine, N,N-dimethylaniline, quinoline, isoquinoline, α-picoline, β-picoline, γ-picoline, quinaldine or pyrimidine.

[0106] I. Compound (1), its salt and synthesis thereof

[0107] The inventors have found in in vitro models, animal models and human clinical trials that ONC201 (compound (1)) has broad anticancer activity, low toxicity, including few (if any) side effects, low genotoxicity and high bioavailability, including oral bioavailability. These features make ONC 201 and various analogs particularly suitable for a variety of applications.

[0108] In one aspect, the present invention provides compound (1): Compound (1) can be prepared by the synthetic method shown in Scheme 1 below.

[0109]

[0110] Solution 1

[0111] In one embodiment, the synthesis of the dihydrochloride salt of compound (1) starts from the commercially available intermediate N-benzyl-3-methoxy-4-piperidone hydrochloride, compound (3). In one embodiment, the synthesis method comprises neutralizing compound (3) with a base (step 1) to produce compound (4), a free radical. In one embodiment, the synthesis method comprises neutralizing compound (3) and an inorganic base to produce compound (4). In one embodiment, the synthesis method comprises neutralizing compound (3) with an organic substrate to produce compound (4). In one embodiment, the intermediate compound (3) is neutralized in the presence of alcohol. For example, the intermediate compound (3) is neutralized in the presence of n-butanol. In one embodiment, the intermediate compound (3) is neutralized in the presence of at least one organic solvent. For example, the intermediate compound (3) is neutralized in the presence of n-butanol and / or ethyl acetate. In one embodiment, the intermediate compound (3) is neutralized in the presence of a base and at least one organic solvent. For example, the intermediate compound (3) is neutralized in the presence of NaHCO3 and n-butanol. In one embodiment, the intermediate compound (3) is neutralized in the presence of n-butanol and triethylamine (Et3N).

[0112] In one embodiment, the synthesis method comprises reacting compound (4) and compound (5) (step 2) to prepare intermediate compound (1). In one embodiment, the reaction in step 2 comprises heating compound (4) and compound (5). In one embodiment, the reaction in step 2 comprises heating compound (4) and compound (5) under reflux in the presence of a solvent. In one embodiment, the reaction in step 2 comprises using a Dean-stark trap to remove water and / or methanol (MeOH) formed in the reaction.

[0113] In one embodiment, the synthesis method comprises forming a dihydrochloride salt of compound (1) (step 3). In one embodiment, the reaction in step 3 comprises treating compound (1) with HCl in dioxane. In one embodiment, the reaction in step 3 comprises treating compound (3) with 4N HCl in dioxane. In one embodiment, the synthesis method optionally comprises recrystallization of the di-salt of compound (1).

[0114] In a preferred embodiment, the synthetic method for preparing the dihydrochloride salt of compound (1) is described in Scheme 2 below.

[0115]

[0116] Solution 2

[0117] II. TNF-related viral-induced ligand ("TRAIL")

[0118] TRAIL protein can be tested in a test sample obtained from a subject to detect TRAIL expression induced by compounds described herein and salts thereof. Immunoassays can be used for TRAIL in test samples, including but not limited to enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunosorbent assay (ELIFA), flow cytometry, immunoblotting, immunoprecipitation, immunohistochemistry, immunocytochemistry, luminescent immunoassay (LIA), fluorescent immunoassay (FIA) and radioimmunoassay. Test methods can be used to obtain qualitative and / or quantitative results. Specific details of suitable test methods for qualitative and quantitative testing of samples are described in standard reference texts, including 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; FM Ausubel et al., Eds., Short Protocols in Molecular Biology, Current Protocols, Wiley, 2002; S. Klussman, Ed., The Aptamer Handbook: Functional Oligonucleotides and Their Applications, Wiley, 2006; Ormerod, MG, Flow Cytometry: a practical approach, OxfordUnivetsity Press, 2000; Givan, AL, Flow Cytometry: first principles, Wiley, New York, 2001; Gorczyca, W., Flow Cytometry in Neoplastic Hematology: morphologic-immunophenotypic eorrelation, Taylor & Francis, 2006; Crowther, JR, The ELISA Guidebook (Methods in Molecular Biology), Humana Press, 2000; Wild, D., TheImmunoassay Handbook, 3. rd Edition, Elsevier Science, 2005, and J.Sambrook andD.W.Russell, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Ptess, 3 rd ed., 2001.

[0119] An example of a method for testing a sample for TRAIL for use in testing the effects of a pharmaceutical composition is described in US Pat. No. 8,673,923 to Wafik S. El-deiry et al., which is incorporated herein by reference in its entirety.

[0120] In one embodiment, the TRAIL test is used to monitor the subject. Thus, for example, before treatment with the pharmaceutical composition, during treatment and / or after treatment, one or more test samples are obtained from the subject to assess the effectiveness of the treatment. In another example, test samples are obtained from the subject at different times to assess the course or progress of the disease or healing. In one embodiment, death receptors can also be analyzed from circulating tumor cells to see if the administration of the compounds described herein or salts thereof increases the amount or type of death receptors.

[0121] Cancers treated using the methods and compositions described herein are characterized by abnormal cell proliferation including, but not limited to, pre-neoplastic hyperproliferation, cancer in situ, tumors and metastasis. The methods and compositions described herein can be used to prevent and improve the signs and / or symptoms of cancer. The terms "treat" and "treatment" for treating a subject's cancer include: preventing, inhibiting or improving a subject's cancer, such as slowing cancer progression and / or reducing or improving signs or symptoms of cancer. Examples of cancers treated using the methods and compositions described herein include, but are not limited to, breast cancer, CNS cancer, colon cancer, ovarian cancer, prostate cancer, leukemia, lung cancer and lymphoma.

[0122] III. Compound (10) and its salt

[0123] In one aspect, provided herein are compounds and related salts of compounds (10) and processes for making the same. One skilled in the art will appreciate that the general principles and concepts described herein in conjunction with compound (1) and its salts (including principles and concepts related to methods and pharmaceutical compositions) are applicable to compounds of formula (10) and their salts.

[0124] In some embodiments, provided herein is compound (10): wherein R1 and R2 independently represent hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, thioaralkyl and acyl groups, wherein when R1 represents CH2Ph, R2 does not represent CH2-((2-CH3)-Ph.

[0125] In some embodiments, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine, C 1-4 Alkylthienyl, C 1-4 Alkylpyridyl, C 1-4 Alkylisoxazolidinyl, C 1-4 Alkylmorpholinyl, C 1-4 Alkylthiazolyl and C 1-4 Alkylpyrazinyl, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine, C 1-4 Alkylthienyl, C 1-4 Alkylpyridyl, C 1-4 Alkylisoxazolidinyl, C 1-4 Alkylmorpholinyl, C 1-4 Alkylthiazolyl, C1-4 The alkylpyrazinyl group is optionally substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In some embodiments, R1 and / or R2 are substituted or unsubstituted, arylalkyl or heteroarylalkyl. In some embodiments, heteroarylalkyl is selected from C 1-4 Alkylpyrrolyl, C 1-4 Alkyl furanyl, C 1-4 Alkylpyridyl, C 1-4 Alkyl-1,2,4-thiadiazolyl, C 1-4 Alkylpyrimidinyl, C 1-4 Alkylthienyl, C 1-4 Alkylisothiazolyl, C 1-4 Alkyl imidazoline, C 1-4 Alkyl tetrazolyl, C 1-4 Alkylpyrazinyl, C 1-4 Alkylpyrimidinyl, C 1-4 Alkylquinolyl, C 1-4 Alkylisoquinolyl, C 1-4 Thioalkylphenyl, C 1-4 Alkylbenzothiophene, C 1-4 Alkyl isobenzofuranyl, C 1-4 Alkylpyrazolyl, C 1-4 Alkyl indolyl, C 1-4 Alkyl purine, C 1-4 Alkylcarbazolyl, C 1-4 Alkylbenzimidazolinyl, C 1-4 Alkylisoxazolyl.

[0126] In some embodiments, R1 and / or R2 are benzyl groups optionally substituted on the benzyl ring with one or more of the following substituents: X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 ,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)ORm ; R m and R n independently selected from hydrogen or C1-C4 alkyl; and wherein p is an integer from 2 to 20 and X represents a halogen, including a fluorine, chlorine, bromine or iodine atom, preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine.

[0127] In some embodiments, R1 is selected from H, CH3, CH2Ph, CH2-(4-CF3-Ph), CH2-(4-F-Ph), CH2-(4-Cl-Ph), CH2-(OCH3-Ph), CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2-(3-thienyl), CH2-2-pyridyl, CH2-4-methyl-2-thiazolyl, CH2-2-pyrazinyl, CH2CH2Ph, CH2CH2(4-N-benzyl-piperazine), CH2-(2,4-diF-Ph), CH2-(3,4-diCl-Ph), CH2-(3,4-diF-Ph), CH2-(3,5-diF-Ph), CH2-((2-CH3)-Ph), CH2CH(OH)Ph, (4-F-Ph)-4-oxobutyl, CH2CH2NHCOOC(CH3)3, CH2CH2CH2NH2 and CD2C6D5. In some embodiments, R2 is selected from H, CH3, CH2Ph, CH2-(4-CF3-Ph), CH2-((2-Cl)-Ph), CH2-((2-F)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2CH2(4-N-benzyl-piperazine), CH2-(2,4-diF-Ph), CH2-(2,4-diF-Ph), CH2-(2,4-diCl-Ph), CH2-(3,4-diCl-Ph), CH2-(3,4-diF-Ph), CH2-(3,5-diF-Ph), CH2-((2-CH3)-Ph), CH2(2-CH3,4-F-Ph), CH2-((4-OCH3)-Ph), CH2-(3- pyridyl), CH2-(3-isoxazolidinyl), CH2CH2-(4-morpholinyl), CH2-(2-F,4-CF3-Ph), CH2CH(OH)Ph, (CH2)3CO-4F-Ph, (4-F-Ph)-4-oxobutyl, CH2CH2NHCOOC(CH3)3, CH2CH2CH2NH2 and CD2C6D5.

[0128] In some embodiments, R1 is hydrogen. In some embodiments, R1 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl group is replaced by C 1-4 Alkyl, C 1-4Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0129] In one embodiment, R2 is a substituted or unsubstituted aralkyl group, such as benzyl or phenethyl. In one embodiment, the aralkyl group is C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, 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 substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In one embodiment, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In one embodiment, heterocycloalkylalkyl or heteroarylalkyl is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted. In one embodiment, the heterocycloalkylalkyl or heteroaralkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0130] In one embodiment, compound (10) has the structure of compound (80): Where R a1 , R a2 , R a3 , R a4 , R a5 , R b1 , R b2 , R b3 , R b4 and R b5 Each is independently selected from X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 ,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R mand C(O)OR m ; R m and R n independently selected from hydrogen or C1-C4 alkyl; and wherein p is an integer from 2 to 20 and X represents halogen.

[0131] In one embodiment, compound (10) has the structure of compound (90) wherein R2 is as defined above, and wherein R b1 , R b2 , R b3 , R b4 and R b5 Each is independently selected from X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 ,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ; R m and R n independently selected from hydrogen or C1-C4 alkyl; and wherein p is an integer from 2 to 20 and X represents halogen.

[0132] In one embodiment, compound (10) has the structure of compound (40) wherein R1 is as defined above, and wherein R a1 , R a2 , R a3 , R a4 and R a5 are each independently selected from hydrogen, 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 , ORm , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ; R m and R n R1 is independently selected from hydrogen or C1-C4 alkyl; and wherein p is an integer from 2 to 20 and X represents a halogen. In one embodiment, R1 is hydrogen. In one embodiment, R1 is a substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In one embodiment, the aralkyl is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, the benzyl group is substituted with one or more halogens. In one embodiment, the benzyl group is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3, -OCH3. In one embodiment, the benzyl group is substituted with one halogen substituent, such as a fluorine substituent, and with two halogen substituents, such as a fluorine substituent, at the two meta positions.

[0133] In one embodiment, compound (40) has the structure of compound (45):

[0134] Where R a1 , R a2 , R a3 , R a4 and R a5 As defined above. In one embodiment, the benzyl group is substituted with one or more halogens. In some embodiments, the benzyl group is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3, and -OCH3. In one embodiment, R a1 or R a5 is halogen, such as fluorine. In one embodiment, R a2 or R a3 is a halogen, such as a fluorine substituent.

[0135] In one embodiment, compound (10) has the structure of compound (50) wherein R1 is as defined above, and wherein R bis selected from X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 ,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ; R m and R n are independently selected from hydrogen or C1-C4 alkyl; and wherein p is an integer from 2 to 20 and X represents halogen, and wherein R a1 R a2 , R a4 and R a5 are each independently selected from hydrogen, X, -CH3, -NO2, -OCH3, -CN, -CXH2, -CX2H, C2-C4 alkyl, -CX3, -CH2(CX3), -CH(CX3)2, -C(CX3)3, -C p X 2p+1 ,-OCX3,-OC p H 2p+1 , -OC p X 2p+1 , OR m , SR m , NR m R n , NR m C(O)R n , SOR m , SO2R m , C(O)R m and C(O)OR m ; R m and R n independently selected from hydrogen or C1-C4 alkyl; and wherein p is an integer from 2 to 20 and X represents a halogen. In one embodiment, R1 is hydrogen. In one embodiment, R1 is a substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In one embodiment, the aralkyl is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C1-4 In one embodiment, R b In one embodiment, one or more R a1 , R a2 , R a4 , R a5 In one embodiment, R a1 , R a2 , R a4 , R a5 is hydrogen, R b In one embodiment, R b is a halogen such as fluorine, and R a1 In one embodiment, R b is fluorine or chlorine, and R a2 is fluorine or chlorine. In one embodiment, R b is CF3. In one embodiment, R b In one embodiment, R b is chlorine and R a1 It's chlorine.

[0136] In one embodiment, compound (50) has the structure of compound (55): Where R a1 , R a2 , R a4 , R a5 , R b As defined above. In one embodiment, R b In one embodiment, one or more R a1 , R a2 , R a4 , R a5 In one embodiment, R a1 , R a2 , R a4 , R a5 is hydrogen, R b In one embodiment, R b is a halogen such as fluorine, R a1 In one embodiment, R b is fluorine or chlorine, R a2 is fluorine or chlorine. In one embodiment, R b is CF3. In one embodiment, Rb is OCH3. In one embodiment, R b is chlorine and R a1 It's chlorine.

[0137] In one embodiment, compound (10) has the structure of compound (60) In some embodiments, R1 is hydrogen. In some embodiments, R1 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, R1 is substituted or unsubstituted heterocycloalkylalkyl or substituted or unsubstituted heteroaralkyl, such as CH2-(2-thienyl), CH2-(3-thienyl), CH2-2-pyridyl, CH2-3-pyridyl, CH2-4-methyl-2-thiazolyl, CH2-2-pyrazinyl, CH2CH2 (4-N-benzyl-piperazine), CH2-(3-isoxazolidinyl) and CH2CH2-(4-morpholinyl). In one embodiment, the aralkyl is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In one embodiment, benzyl is substituted with one or more halogens. In some embodiments, benzyl is substituted with one or more substituents selected from the following: halogen (e.g., fluorine) -CH3, -CF3, -OCH3. In one embodiment, the substituents at the para position of benzyl are selected from halo, -CH3, -CF3, -OCH3. In one embodiment, R1 is fluorophenyloxybutyl or hydroxyphenylethyl.

[0138] Scheme 3 describes the synthesis of compounds of formula (10)

[0139]

[0140] Solution 3

[0141] Compounds of formula (10) are synthesized starting from substituted piperidones which are converted by reaction with substituted aminoimidazolines to give the core compound (10). There are two routes where the R1 substituent is present in the piperidone (e.g. 68). In this route, (80) is reacted with dimethyl carbonate at 80°C using sodium hydride in toluene to form the piperidone ester (69). The commercially available methylthiohydrazoline HI salt (63) is reacted with an amine in dioxane at 70°C to give the R2-substituted amino-imidazoline (64) as its HI salt. The reaction of (64) with the piperidone ester (69) in 1-butanol with removal of water over 3-6 hours via a Dean-Stark trap gives the trichloro compound (10). In a variation of this scheme, the N-BOC protected piperidone (61) is converted to the BOC protected compound (65) by the same method, which is treated with HCl in dioxane to remove the BOC group and then converted to (66) extracted with 1 N NaOH and methyl chloride. Subsequent treatment of (66) with a halide (67) or epoxide (70) affords the desired compound (10).

[0142] The crude product can be purified by column chromatography eluting with methyl chloride methanol, or by HPLC using acetonitrile:TFA:H2O to give the final product as a free base or a TFA salt. Treatment of the free base with HCl in dioxane, or lyophilization of the TFA salt yields product (10) as an HCl or TFA salt. Alternatively, the free base can be treated with another inorganic or organic acid to form other salts, typically selected from salts known to be pharmaceutically acceptable salts. Salts of compound (10) are typically solids, and examples have been crystallized from ethanol or other solvents to give high quality crystals. In the case of compound (1), the tricyclic structure was determined by X-ray crystallography and NMR.

[0143] 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 it in the cellular environment. Expression of these binding targets can be used to predict response to compound (1) (ONC201) or its analogs (i.e., as biomarkers). In addition, these compounds can be used to screen for structurally unrelated anticancer compounds known in the art in competition assays to identify drugs that can surpass the target interaction with higher affinity. In addition, these molecules can have drug properties that produce therapeutic improvements or allow additional therapeutic applications by changing drug properties, including but not limited to pharmacokinetics, efficacy, safety, biodistribution or metabolism.

[0144] Examples of Compound (10)

[0145]

[0146]

[0147] As described in Example 12 below, compound (1) with an aminoalkyl linker (i.e., compound (33)) was used to identify proteins that interact with compound (1). Compound (1) was found to interact with proteins involved in N6-methyl-adenosine (m 6 A) Protein interactions of mRNA methylation. 6 A Proteins that epigenetically modify mRNA include those listed in Table 1. These proteins include methylated mRNAs (RNA writers), such as METTL3, METTL14, WTAP, and KIAA1429; those that demethylate mRNAs (RNA writers), such as METTL3, METTL14, WTAP, and KIAA1429; 6 A mRNA (RNA wiper), such as FTO and ALKBH5; and specific recognition of m 6 A RNA (RNA reader), such as YTHDF3, YTHDF2, YTHDF1, YTHDC1 and YTHDC2.

[0148] Table 1: m 6 A mRNA methylation protein

[0149]

[0150] IV.x Evaluate the sensitivity and efficacy of treatment options

[0151] Measuring expression, post-translational modification, or activity level, or mutation in eIF2-α, ATF4, CHOP, DR5 or cleaved or whole cytokeratin 18 can be used to predict response or sensitivity, to identify subjects who may respond to treatment methods described herein, such as treatment methods with compound (1) or a pharmaceutically acceptable salt thereof. In addition, measuring expression, post-translational modification, or activity level, or mutation in eIF2-α, ATF4, CHOP, DR5 or cleaved or whole cytokeratin 18 can be used to evaluate or monitor the effectiveness of treatment methods described herein. In addition, measuring expression, post-translational modification, or activity level, or mutation in eIF2-α, ATF4, CHOP, DR5 or cleaved or whole cytokeratin 18 can be used to screen for structurally unrelated anticancer compounds in vitro or in vitro. For example, competition and other assays can be used to identify drugs that can exceed the target interaction with higher affinity to compare the changes in these levels with the respective changes produced by compound (1) or its analogs. Testing can also be performed on living mammalian cells, which more closely approximates the effects of a drug at a specific serum level in vivo, or on microsomal extracts prepared from cultured cell lines.

[0152] In some embodiments, the subject has cancer or is at risk of cancer. In some embodiments, the treatment regimen comprises administering an effective amount of compound (1) or compound (10) or an analog thereof. In some embodiments, the treatment regimen comprises administering an effective amount of compound (1). In some embodiments, the treatment regimen comprises administering an effective amount of a compound of formula (10). In some embodiments, the compound of formula (10) is a compound of formula (40), for example, a compound of formula (45). In some embodiments, the compound of formula (10) is a compound of formula (50), for example, a compound of formula (55). In some embodiments, the compound of formula (10) is a compound of formula (80). In some embodiments, the compound of formula (10) is a compound of formula (90). In some embodiments, the compound of formula (10) is a compound of formula (60). In some embodiments, the analog of compound (1) has a molecule selected from compound (25), compound (26), compound (27), compound (28), compound (29), compound (30), or compound (31).

[0153] The level of the predetermined standard can be, for example, the mean or median level measured in a sample from a subject. The level of the predetermined standard can be measured under the same or substantially similar experimental conditions as measuring a sample from a subject. The level of the predetermined standard can be obtained from a subject to which a chemical substance (1) or compound (10) or its analog is reacted. In one embodiment, the predetermined standard is obtained from a subject who is responsive to chemical treatment, and if the level in the sample from the standard is similar to the level in the standard, the subject can be classified as having a possibility of responding to treatment. The level of the predetermined standard can be obtained from a subject who is unresponsive to chemical treatment. In one embodiment, the predetermined standard is obtained from a subject who is unresponsive to compound treatment, and if the level in the subject sample is different from the level in the expected standard (e.g., up-regulated or down-regulated), the subject can be classified as having a possibility of responding to treatment. The level of the predetermined standard can be obtained from a normal healthy subject.

[0154] Immunoassays can be used to measure protein or methylation levels in a sample, including but not limited to enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunosorbent assay (ELIFA), flow cytometry, immunoblotting, immunoprecipitation, immunohistochemistry, immunocytochemistry, luminescent immunoassay (LIA), fluorescent immunoassay (FTA), and radioimmunoassay. A Methyl chemical levels in mRNA samples can be obtained by methyl RNA immunoprecipitation (Me-RIP) or other quantitative biochemical assays known in the art.

[0155] Nucleic acid mutations can be determined by any of many known methods. For example, a personal biological sample can be obtained. This biological sample includes but is not limited to body (such as urine, saliva, plasma or serum) or tissue sample (such as cheek tissue sample or cheek cell). Then the biological sample can be sequenced or scanned using known methods. For example, DNA arrays can be used to analyze at least a portion of the subject's genomic sequence. In addition, all or part of the genomic sequence information can be used. Such sequences can be determined using standard sequencing methods including chain termination (Sanger dideoxynucleotides), dye terminator sequencing and SOLID TM sequencing (Applied Biosystems). The entire genomic sequence can be sequenced by restriction enzyme cutting or shearing (mechanical) into shorter fragments. Known methods such as PCR and vector-based cloning methods (such as Escherichia coli) can also be used to amplify DNA sequences. In one embodiment, at least a portion of the subject's genetic material (such as DNA, RNA, mRNA, cDNA, other nucleotide bases or derivatives of these) is scanned or sequenced using, for example, a conventional DNA sequencer or chip-based technology to determine whether there is a mutation or copy number variation.

[0156] In one aspect, provided herein is a method for identifying and treating a subject with a condition and can respond to the treatment regimen described herein. In one embodiment, the method includes (i) identifying whether a subject with the condition can respond to the treatment regimen described herein; and (ii) treating a subject that may respond to the treatment regimen with the treatment regimen. In one embodiment, the subject has cancer or is at risk of cancer. In one embodiment, the treatment regimen includes administering an effective amount of compound (1) or compound (10) or an analog thereof. In some embodiments, the treatment regimen includes administering an effective amount of compound (1). In some embodiments, the treatment regimen includes administering an effective amount of a compound of formula (10). In some embodiments, the compound of formula (10) is a compound of formula (40), such as a compound of formula (45). In some embodiments, the compound of formula (10) is a compound of formula (50), such as a compound of formula (55). In some embodiments, the compound of formula (10) is a compound of formula (80). In some embodiments, the compound of formula (10) is a compound of formula (90). In some embodiments, the compound of formula (10) is a compound of formula (60). In some embodiments, the analog of compound (1) has a structure selected from the structure of compound (25), compound (26), compound (27), compound (28), compound (29), compound (30), or compound (31).

[0157] The level of the predetermined standard can be, for example, the mean or median level measured in a sample from a subject. The level of the predetermined standard can be measured under the same or substantially similar experimental conditions as measuring a sample from a subject. The level of the predetermined standard can be obtained from a subject to which a chemical substance (1) or compound (10) or its analog is reacted. In one embodiment, the predetermined standard is obtained from a subject who is responsive to compound treatment, and if the level of the subject's sample is similar to the level in the standard, the subject can be classified as having a responsiveness treatment. The level of the predetermined standard can be obtained from a subject who is unresponsive to chemical treatment. In one embodiment, the predetermined standard is obtained from a subject who is unresponsive to compound treatment, and if the level in the subject's sample is different from the level in the expected standard (e.g., up-regulated or down-regulated), the subject can be classified as having a possibility of responding to treatment. The level of the predetermined standard can be obtained from a normal healthy subject. Immunoassays can be used to measure the protein level in a sample.

[0158] In one aspect, provided herein is a method for treating and evaluating the effectiveness of a treatment of a subject having a condition. In some embodiments, the method comprises (i) treating the subject according to the treatment methods described herein; (ii) evaluating the effect of the treatment as described herein. In some embodiments, the subject has cancer or is at risk of cancer. In some embodiments, the treatment regimen comprises administering an effective amount of compound (1) or compound (10), or an analog thereof. In some embodiments, the treatment regimen comprises administering an effective amount of compound (1). In some embodiments, the treatment regimen comprises administering an effective amount of a compound of formula (10). In some embodiments, the compound of formula (10) is a compound of formula (40), such as a compound of formula (45). In some embodiments, the compound of formula (10) is a compound of formula (50), such as a compound of formula (55). In some embodiments, the compound of formula (10) is a compound of formula (80). In some embodiments, the compound of formula (10) is a compound of formula (90). In some embodiments, the compound of formula (10) is a compound of formula (60). In some embodiments, the analog of compound (1) has a structure selected from the structure of compound (25), compound (26), compound (27), compound (28), compound (29), compound (30), or compound (31).

[0159] Other conditions that may be amenable to the methods described herein include, but are not limited to, attention deficit disorder, addiction, epilepsy, viral infection, inflammation, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, cardiovascular diseases such as coronary artery disease, cardiomyopathy, hypertensive heart disease, heart failure, cor pulmonale, arrhythmias, inflammatory heart disease, endocarditis, inflammatory heart disease, myocarditis, valvular heart disease, cerebrovascular disease, peripheral arterial disease, congenital heart disease, rheumatic heart disease, diabetes, and light chain amyloidosis.

[0160] V. Composition

[0161] In one aspect, a pharmaceutical composition is provided, comprising formula (10): Or formula (1): Compounds and their pharmaceutically acceptable salts. In one embodiment, the pharmaceutical composition comprises a pharmaceutically acceptable salt of the compound. In one embodiment, the salt is a pharmaceutically acceptable monosalt of the compound. In one embodiment, the salt is a pharmaceutically acceptable disalt of the compound. In one embodiment, the salt is a pharmaceutically acceptable monosalt or polysalt (e.g., disalt or trisalt) selected from hydrochloride, hydrobromide, bisulfate, sulfate, phosphate, fumarate, succinate, oxalate and lactate, bisulfate, hydroxyl, tartrate, nitrate, citrate, tartrate, carbonate, malate, maleate, fumarate sulfonate, methanesulfonate, formate, acetate and carboxyl. In one embodiment, the salt is a pharmaceutically acceptable salt selected from p-toluenesulfonate, benzenesulfonate, citrate, methanesulfonate, oxalate, succinate, tartrate, fumarate and maleate. In one embodiment, the salt is a pharmaceutically acceptable salt selected from ammonium, sodium, potassium, calcium, magnesium, zinc, lithium and / or with counterions such as methylamino, dimethylamino, diethylamino and triethylamino counterions. In one embodiment, the salt is a dihydrochloride or dihydrobromide salt of the compound.

[0162] Compound (1) has the same chemical structure as revealed by structural analysis (eg, NMR, X-ray diffraction) of chemical substance NSC350625 obtained from the National Cancer Institute's Developmental Therapeutics Program repository.

[0163] In one embodiment, the pharmaceutical composition includes a di-salt (e.g., dihydrochloride) of compound (1) or an analog thereof (e.g., a compound of formula (10)). Salts (e.g., di-salts or tri-salts) of analogs of compound (1) can be prepared from analogs of compound (1), which can be synthesized as described herein, or using standard chemical synthesis methods known to those of ordinary skill in the art.

[0164] In one embodiment, the pharmaceutical composition includes at least one pharmaceutically acceptable carrier. Suitable pharmaceutically acceptable carriers include, but are not limited to, those described 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, methods of preparing drug compositions and various dosage forms and modes of administration are well known in the art and are described, for example, in detail in Pharmaceutical Dosage Forms: Tablets, edited by Larry L. Augsburger & Stephen W. Hoag., London: Informa Healthcare, 2008; and in LV Allen, Jr. et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 8th Ed., Philadelphia, Pa.: Lippincott, Williams & Wilkins, 2004; AR Gennaro, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st ed., 2005, particularly chapter 89; and JG Hardman et al., Goodman & Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill Professional, 10th ed., 2001.

[0165] In one embodiment, the pharmaceutical composition is for ocular administration. In one embodiment, the pharmaceutical composition is for topical administration. In one embodiment, the pharmaceutical composition is formulated as oil drops, ointments or liquids. In one embodiment, the pharmaceutical composition includes conventional pharmaceutical carriers such as aqueous, powdered or oily bases, thickeners, etc.

[0166] In some embodiments, the pharmaceutical composition is a preparation for intravenous administration. In one embodiment, the intravenous preparation comprises a compound of formula (10) or a pharmaceutically acceptable salt thereof dissolved in a solvent. In one embodiment, the solvent comprises water. In one embodiment, the intravenous preparation comprises a 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 preparation comprises a 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 preparation comprises about 0.005% 0.05%, or about 0.5% to about 0.1%, 1% or 10% of a compound or a salt thereof. In one embodiment, the intravenous preparation comprises about 0.05%, 0.5% or 5% of a compound or a salt thereof. In some embodiments, the intravenous preparation comprises a higher or lower concentration of a compound or a salt thereof.

[0167] In one embodiment, the pH of the intravenous formulation is about 3. In one embodiment, the intravenous formulation is adjusted to pH 3 with a phosphate buffer. . In one embodiment, the intravenous formulation includes glucose or sodium chloride. In one embodiment, the intravenous formulation includes the compound or its salt at a concentration of about 5 mg / mL and pH 3, forming a stable solution. In one embodiment, the intravenous formulation includes the compound or its salt at a concentration of about 5 mg / mL, pH <5, forming a stable solution. In one embodiment, the intravenous formulation includes the compound or its salt and one or more antioxidants. In one embodiment, the intravenous formulation includes a mixture of mono- and dihydrochloride salts of the compound. In one embodiment, the intravenous formulation contains the compound or its salt at a concentration of about 10 mg / mL of a 1% solution, 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.

[0168] In one embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. In one embodiment, a suitable pharmaceutically acceptable carrier comprises oil. In one embodiment, a suitable pharmaceutically acceptable carrier comprises sterile water. In one embodiment, a suitable pharmaceutically acceptable carrier comprises an aqueous carrier. In some embodiments, the intravenous formulation comprises glucose and / or sodium.

[0169] In one embodiment, the intravenous formulation comprises compound (1) or its analog or dihydrochloride dissolved in 25 mg / mL of water. In one embodiment, the intravenous formulation is adjusted to pH 3 with phosphate buffer. In one embodiment, the intravenous formulation comprises glucose or sodium chloride. In one embodiment, the intravenous formulation comprises a dihydrochloride of compound (1) or its analog at a higher or lower concentration. In one embodiment, the intravenous formulation comprises compound (1) or its analog or dihydrochloride at a concentration of about 5 mg / mL. In one embodiment, the intravenous formulation comprises compound (1) or its analog or dihydrochloride at a concentration of about 5 mg / mL and pH 3 to form a stable solution. In one embodiment, the intravenous formulation comprises compound (1) or its analog or dihydrochloride at a concentration of about 5 mg / mL and pH <5 to form a stable solution. In one embodiment, the intravenous formulation comprises compound (1) or its analog or dihydrochloride and one or more antioxidants. In one embodiment, the intravenous formulation comprises a mixture of mono- and dihydrochloride salts of compound (1) or its analog. In one embodiment, the intravenous formulation comprises Compound (1) or an analog thereof or its dihydrochloride salt as a 1% solution at a concentration of about 10 mg / mL, for example, the intravenous formulation has a pH of about 3.3. In one embodiment, the pH is less than 4.0.

[0170] In one embodiment, the intravenous formulation comprises about 0.5% to about 10% (or about 5 mg / mL to about 100 mg / mL) of compound (1) or its analog or its di-salt. In one embodiment, the intravenous formulation comprises about 5% (or about 50 mg / mL) of compound (1) or its analog or its di-salt. In one embodiment, the intravenous infusion rate may be slowed down to reduce the side effects of compound (1) or its analog or its di-salt.

[0171] In one embodiment, the pharmaceutical composition comprises about 0.1-99% of a salt of compound (1) or its analog; and a pharmaceutically acceptable carrier, such as oil or sterile water or other aqueous carrier. In one embodiment, the pharmaceutical composition comprises a mono- or di-salt of compound (1) or its analog in an oral dosage form in the range of about 5% to about 50%.

[0172] In some embodiments, the pharmaceutical composition includes an antioxidant. Suitable antioxidants include: ascorbic acid derivatives, such as ascorbic acid, isoascorbic acid, sodium ascorbate, thioglycerol, cysteine, acetylcysteine, cystine, dithioerythritol, dithiothreitol, glutamic acid, tocopherol, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), Sulfites such as sodium sulfate, sodium bisulfite, acetone sodium bisulfite, sodium metabisulfite, sodium sulfite, sodium formaldehyde sulfoxylate and sodium thiosulfate, dihydroguaiaretic acid. It should be noted that antioxidants for aqueous preparations generally include: sodium sulfite, sodium metabisulfite, Sodium formaldehyde sulfonate and ascorbic acid and combinations thereof, and antioxidants for oil-based solutions, organic solvents include butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA) and propyl propionate and combinations thereof. In other embodiments, the antioxidant can be one or more of flavonoids, isoflavones, monothioglycerol, L-cysteine, thioglycolic acid, alpha-tocopherol, ascorbyl 6-palmitate, dihydrolipoic acid, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), vitamin E, propyl gallate, beta-carotene, ascorbic acid. Antioxidants can generally be used at about 0.1% to 1.0% by weight, more typically about 0.2%.

[0173] In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10), or an analog thereof or a pharmaceutically acceptable salt thereof, and at least one other therapeutic agent. For example, the other therapeutic agent is selected from hormone analogs and anti-hormones, aromatase inhibitors, LHRH agonists and antagonists, growth factor inhibitors, growth factor antibodies, growth factor receptor antibodies, tyrosine kinase inhibitors; antimetabolites; antitumor antibiotics; platinum derivatives; alkylating agents; antimitotic agent S; tubuline inhibitors; PARP inhibitors, topoisomerase inhibitors, serine / threonine kinase inhibitors, tyrosine kinase inhibitors, protein-protein interaction inhibitors, RAF inhibitors, MEK inhibitors, ERK inhibitors, IGF-1R inhibitors, ErbB receptor inhibitors, rapamycin analogs, BTK inhibitors, CRM1 inhibitors (e.g., KPT185), P53 modulators (e.g., Nutlins), anti-angiogenic factors (e.g., atilibinin, afliposome, sorafenib and regorafenib), amifostine, anagrelide, clodronate, filgrastim, interferon, interferon α, folinic acid, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate and porphyrin, 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-alethine, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A 105972, A 204197, abiraterone, aldesleukin, ivermectin, alloectin-7, altret amine, alvocidib, amonafide, anthrapyrazole, AG-2037, AP-5280, apaziquone, apomine, aripiprazole, atroxifen, atamitane, atestamitan, aspirin PE, AVLB, AZ10992, ABX-EGF, AMG-479 (ganituzumab), ARRY 162, ARRY 438162, ARRY-300, ARRY-142886 / AZD-6244 (selumetinib) 704 / AZD-8330, AR-12, AR-42, AS-703988, AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026 (primasertib) AZD-2014, azacytidine, azabetaloxone B, azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, disodium docosanoate, BCX-1777, BKM-120, bucomitin, BLP-25,BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW2992 (afatinib, tomtovok), BIBF1120 (vargatef), BI 836845, BI 2536,BI 6727,BI 836845,BI 847325,BI 853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryostatin-1, bortezomib, brostallicin, busulphan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, capecitabine, canfosfamide, capecitabine, carboxyphthalimin, CCl-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, cefotaxime, ceftriaxone, celecoxib, celmoleukin, cemadotin, CH4987655 / RO-4987655, chlortriphenylamine, cilengitide, cyclosporine, CDA-II, CDC-394, CKD-602, CKI-27, clofarabin, colchicine, combretastatin A4, COT inhibitors, CHS-828, CH-5132799, CLL-Thera, CMT-3cryptophycin52, CTP-37, CTLA-4 monoclonal antibody, CP-461, CV-247, cyanomorpholino doxorubicin, cytarabine, D 24851, decitabine, deoxyvitamin, deoxyvitamin, deoxyribonucleic acid, dephospholipid ascorbate B, dexamethasone, dexamethasone, diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10, doranidazole, DS-7423, E7010, E-6201, edatrexat, edotreotide, efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509, enzastaurin, itromycin B, epothilone, epothilone B, epothilone, ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine, ethinyl estradiol, exatecan, exatecan mesylate, exemestane, exisulind, fenretinide, filgretan, filgrolidon, folic acid, FOLFOX, FOLFOX4, FOLFIRI, gefitinib, gemtuzumab, gefitinib, gefitinib, gefitinib, GFC-100,GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100, gp100 peptide vaccine, GSK-5126766, GSK-690693, GSK-1120212 (trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron, Herceptin, hexamethylmelamine, histamine, homoharrington nine, hyaluronic acid, hydroxyurea, hydroxyprogesterone caproate, ibandronate, ibrutinib, itraconazole, itrastatin, escitalopram, IDN-5109, IGF-1R inhibitor, IMC-1C11, IMC-A12 (cixutumumab), immunoglobulin, hemolysin, interferon alpha-2a, interferon alpha-2b, pegylated interferon alpha-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafamib, ipilimumab, isoflavone, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, kahalid F, ketoconazole, KW-2170, KW-2450, lobaplatin, leflunomide, isoflavone plus, leuprorelin, leuporelin, lexidronam, LGD-1550, linezolid, THB, tylosin, lorestat, ruxolitin, LU 223651, lurtotecan, LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechlorethamine, MEK inhibitors, MEK-162, methyltestosterone, methylprednisolone, MEDI-573, MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, midostaurin, minodronic acid, mitomycin, mivobulin, MK-2206, MK-0646 (dalotuzumab), MLN518, motexafin gadolinium, MS-209, MS-275, MX6, neridronate, neratinib, Nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nolatrexed, norrelin, N-acetylcysteine, 06-benzylguanine, oblimersen, Omeprazole,oncophage, oncoVEXGM-CSF, ormiplatin, ortataxel, OX44 antibody, OSI-027, OSI-906 (1insitinib), 4-1BB antibody, oxantrazole, estrogen, panitumumab, patupilone, pegfilgrastim, PCK-3145, pEG-005, PF-05197281, PF-05212384, PF-04691502, PHT-427, P-04, PKC412, P54, PI-88, pelitinib, pemetrexed, pentrix, perifosine, perillylalcohol, pertuzumab, PI3K inhibitor, PI3K / mTOR inhibitor, PG-TXL, PG2, PLX-4032 / RO-5185426 (vemurafenib)-3603 / RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pivaloyloxymethylbutyrate, heptameric isoprenone, phenoxodiol O, PKI166, plevitrexed, mithramycin, polymalonic acid, porphomicin, prednisone, prednisolone, quinolone, R115777, RAF-265, ramosetron, ranpimase, RDEA-119 / BAY869766, RDEA-436, rebeccamycin analogs, receptor tyrosine kinase (RTK) inhibitors, regorafenib, revimid, RG-7167, RG-7304, RG-7421, RG-7321, RG 7440, rhizobactin, rhu-MAb, relifurcin, risedronate, rituximab, rofanumab, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR109881A, naringenone, erythromycin, R-flurbiprofen, RX-0201, S-9788, abalabicin, SAHA, sargramostim, satraplatin, SB 408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustin, 4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spirocarboplatin, keratinine, simvastatin hydroxamic acid, sutent, T 900607, T 138067, TAK-733, TAS-103, tylosin, taparopenem, tylosin, tenosartan, talysaptan, tegafur, temozolomide, tesarofin, temofil, temofilin,testosterone, propionate, tesmid, tetrachloroplatinic acid, tetrodotoxin, tizanpine, thalidomide, trastuzumab, tirazole, trovastatin, trovastatin, tramatinib, TransMID-107, retinoic acid, trastuzumab, tremelimumab, tretinoin, triacetyluridine, triapine, triciribine, trimethoprim, TLK-286TXD 258, tykerb / tyverb, urobilirubin, psychrin, varasini, vincristine, virulizin, WX-UK1, WX-554, vectibix, xeloda, XELOX, XL-147, XL-228, XL-281, XL-518 / R-7420 / GDC-0973, XL-765, YM-511, YM-598, ZD-4190, ZD-6474, ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZD1839, ZSTK-474, zoledronat, zosuquidar and combinations thereof. ,

[0174] In one embodiment, the other therapeutic agent comprises a hormone analog, an antihormone or is selected from tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate, flutamide, nilutamide, bicalutamide, aminoglutarimide, cyproterone acetate, finasteride, buserelin acetate, fludrocortisone, in one embodiment, the other therapeutic agent comprises one or more LHRH agonists and / or antagonists selected from goserelin acetate, cloproceptin acetate, triptorelin pamoate and combinations thereof and wherein the LHRH antagonist is selected from gagarelix, cetrorelix, abalex, ozarelin, gagarelix combination. In one embodiment, the other therapeutic agents include one or more growth factor inhibitors selected from the group consisting of inhibitors of platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), insulin-like growth factor (IGF), human epidermal growth factor (HER) and hepatocyte growth factor (HGF). In one embodiment, the other therapeutic agents include one or more inhibitors of human epidermal growth factor HER2, HER3 and HER4. In one embodiment, the other therapeutic agents include one or more tyrosine kinase inhibitors selected from the group consisting of cetuximab, gefitinib, imatinib, lapatinib and trastuzumab and combinations thereof. In one embodiment, the other therapeutic agents include one or more aromatase inhibitors selected from the group consisting of anastrozole, letrozole, liarozole, vorozole, exemestane, atamestane and combinations thereof. In one embodiment, the other therapeutic agents include one or more antimetabolites, which are antifolates selected from methotrexate, raltitrexed and pyrimidine analogs. In one embodiment, the other therapeutic agents include one or more antimetabolites which are pyrimidine analogs selected from 5-fluorouracil, capecitabine and gemcitabine. In one embodiment, the other therapeutic agents include one or more antimetabolites which are purine and / or adenosine analogs selected from mercaptopurine, thioguanine, cladribine and pentostatin, cytarabine, fludarabine and combinations thereof. In one embodiment, the other therapeutic agents include one or more antineoplastic drugs, doxorubicin, daunomycin, epirubicin and desmethoxydaunorubicin, scleromycin-C, bleomycin, actinomycin D, mithramycin, streptozocin and combinations thereof. In one embodiment, the other therapeutic agents comprise 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 estramustin, mecloreth amine, melphalan, chlorambucil, busulphan, dacarbazin, cyclophosphamide,ifosfamide, temozolomide, nitsoureas and combinations thereof. In one embodiment, the other therapeutic agents include nitsoureas selected from carmustin, lomustin, thiotepa and combinations thereof. In one embodiment, the other therapeutic agents include antimitotic agents selected from vinca alkaloids and taxanes. In one embodiment, the other therapeutic agents include one or more taxanes selected from paclitaxel, docetaxel and combinations thereof. In one embodiment, the other therapeutic agents include one or more vinblastine alkaloids selected from vitamins, verinoxil, verinrein, vincristine and combinations thereof. In one embodiment, the other therapeutic agents include one or more topoisomerase inhibitors that are epipodophyllotoxins. In one embodiment, the other therapeutic agents include one or more epipodophyllotoxins selected from etoposide and fenbifu, teniposide, amsacrin, topotecan, irinotecan, mitoxantron and combinations thereof. One or more serine / threonine kinase inhibitors are selected from PDK 1 inhibitors, B-Raf inhibitors, mTOR inhibitors, mTORC1 inhibitors, PI3K inhibitors, dual mTOR / PI3K inhibitors, STK 33 inhibitors, AKT inhibitors, PLK 1 inhibitors, CDK inhibitors, Aurora kinase inhibitors and combinations thereof. In one embodiment, the other therapeutic agents comprise one or more tyrosine kinase inhibitors, which are PTK2 / FAK inhibitors. In one embodiment, the other therapeutic agents comprise 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, sirolimus, ridaforolimus, sirolimus and combinations thereof. In one embodiment, the other therapeutic agents include one or more therapeutic agents selected from amifestine, anagrelide, clodronat, filgrastin, interferon, interferon alpha, leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate and porfimer and combinations thereof. In one embodiment, the additional therapeutic agents include one or more therapeutic agents selected from 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-alethine, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A 105972, A 204197, abiraterone, aldesleukin, acitretin, allovectin-7, altretamine, alvcidib, amonafide, anthrapyrazole, AG-2037,AP-5280, azaquinone, aritol, alicandine, atroxifen, atamistatin, artestestane, aspirin PE, AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), ARRY 162, ARRY 438162, ARRY-300, ARRY-142886 / AZD-6244 (xelometine)-704 / AZD-8330, AR-12, AR-42, AS-703988, AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026 (primasertib), AZD-2014, azacytidine, azaprotectin B, azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, disodium docosanoate, BCX-1777, BKM-120, bucomitin, BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW 2992 (afatinib, tomtovok), BIBF 1120 (vargatef), BI 836845, BI 2536, BI 6727, BI 836845, BI 847325, BI 853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryostatin-1, bortezomib, brostallicin, busulphan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, capecitabine, canfosfamide, capecitabine, carboxyphthalimin, CCl-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, cefotaxime, ceftriaxone, celecoxib, celmoleukin, cemadotin, CH4987655 / RO-4987655, chlortriphenylamine, cilengitide, cyclosporine, CDA-II, CDC-394, CKD-602, CKI-27, clofarabin, 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, cyanomorpholino doxorubicin, cytarabine,D 24851, decitabine, deoxyrubicin, desogestrel, dexamethasone, dexamethasone, diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10, dorazolidinone, DS-7423, E7010, E-6201, edatrexamine, eflornithine, EGFR inhibitors, EKB-569, EKB-509, enzastaurin, enzalutamide, edatrexamine, epothilone B, epratuzumab, ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine, ethinyl estradiol, exatecan, exatecan mesylate, exemestane, exisulind, fenretinide, fenacic acid, floxetine, folic acid, FOLFOX, FOLFOX4, FOLFIRI, mesultan, flutoxetine, camptothecin, maltosaccharide, gefinitib, gemtuzumab, gimatecan, glucosamide, GCS-100, GDC-0623, GDC-0941 (pictrelisib) 0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100, gp100-peptide vaccine, GSK-5126766, GSK-690693, GSK-1120212 (trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron, Herceptin, hexamethylmelamine, histamine, homoserine, hyaluronic acid, hydroxyurea, hydroxyprogesterone caproate, ibandronate ibrutinib, itrafenib, isostearol, IDN-5109, IGF-1R inhibitor, IMC-1C11, IMC-A12 (cixutumumab), immunoglobulin, hemolysin, interferon α-2a, interferon α-2b, pegylated interferon α-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib, isoproterenol, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, kahalid F, ketoconazole, KW-2170, KW-2450, lopaplatin, leflunomide, leuporelin, lexidronam, LGD-1550, linezolid, lutetium tuxaphyrin, lomotrexol, loosxantrone, LU 223651, lurtotecan,LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechlorethamine, MEK inhibitor, MEK-162, methyltestosterone, methylprednisolone, MEDI-573, MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, midostaurin, minodronic acid, mitomycin, mivobulin, MK-2206, MK-0646 (dalotuzumab), MLN518, motexat in gadolinium, MS-209, MS-275, MX6, neridronate, neratinib, Nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nolatrexine, norethin, N-acetylcysteine, 06-benzylguanine, oblimersen, omeprazole, oncophage, oncoVEXGM-CSF, ormiplatin, ortataxel, OX44 antibody, OSI-027, OSI-906 (linsitinib), 4-1BB antibody, oxantrazole, estrogen, panitumumab, 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, perillylalcohol, pertuzumab, PI3K inhibitor, PI3K / mTOR inhibitor, PG-TXL, PG2, PLX-4032 / RO-5185426 (vemurafenib), PLX-3603 / RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pivaloyloxymethylbutyrate, heptapolyvidone, phenoxydiol O, PKI166, pertussis, mithramycin, polylysine, porphomicin, prednisone, prednisolone, quinine, quinupristin, R115777, RAF-265, ramosetron, rapamycin, RDEA-119 / BAY 869766, RDEA-436, Rebeccamycin analogs, Receptor tyrosine kinase (RTK) RG-7167, RG-7304, RG-7421, RG-7321, RG 7440,rhizobactin, rhu-MAb, rinfabate, risedronic acid, rituximab, robatumumab, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR 109881A, pyridazine, erythromycin, R-flurbiprofen, RX-0201, S-9788, doxorubicin, SAHA, sargramostim, satraplatin, SB 408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustin, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spirocycline, keratinine, simvastatin hydroxamic acid, sutent, T 900607, T 138067, TAK-733, TAS-103, tacicaine, tacaroline, tylosin, tasadone, tegafur, temozolomide, tesarofen, testosterone, testosterone propionate, tesmefen, tetrachloroplatinic acid, tetrodotoxin, tezazepine, thalidomide, trastuzumab, enalapril, tolmetib, tetrazodone, tovastatin, tolmitre, tomifenfen, tramatinib, TransMID-107, retinoic acid, trastuzumab, tremelimumab, tretinoin, triacetyluridine, triaziridine, TLK-286TXD 258, tykerb / tyverb, urocidin, valrubicin, vatalanib, vincristine, vitamin, virulizin, WX-UK1, WX-554, vectibix, xeloda, XELOX, XL-147, XL-228, XL-281, XL-518 / R-7420 / GDC-0973, XL-765, YM-511, YM-598, ZD-4190, ZD-6474, ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZD1839, ZSTK-474, zoledronat, zosuquidar and combinations thereof. ,

[0175] In one embodiment, the other therapeutic agent comprises a steroid. Steroids include, but are not limited to, dexamethasone, prednisolone, methylprednisolone, prednisone, hydrocortisone, triamcinolone acetonide, betamethasone and corticosteroids. In one embodiment, the other therapeutic agent comprises an antiemetic. Dopamine agonists (e.g. domperidone, olanzapine, haloperidol, haloperidol, chlorpromazine, chlorpromazine, alizapride, atorvastatin, etc.), 5-HT 3 receptor agonists (e.g., doxacetron, granisetron, ondansetron, tropisetron, proclopramide and metoclopramide), NK1 receptor antagonists (e.g., aspirin and placebo), antihistamines (e.g., cimetidine, diphenylamine, dimethylbenzidine, doxylcycline, medilide, promethazine, hydroxyzine), cannabinoids (e.g., cannabis, tanshinone and sativex), benzodiazepines (e.g., midazolam and lorazepam), anticholinergics (e.g., hyoscine), triphenylether carboxamide, ginger, emeffol, propofol, mint, thymol and ferulic acid.

[0176] In one embodiment, the other therapeutic agent comprises an anticancer agent, including a mitotic inhibitor. In one embodiment, the mitotic inhibitor comprises a taxane. In one embodiment, the mitotic inhibitor comprises a taxane selected from paclitaxel and docetaxel.

[0177] In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or an analog thereof, or a pharmaceutically acceptable salt thereof; and at least one anticancer agent, including but not limited to one or more of avermectin, aclarubicin, atradazole, aksunin, atifostine, acitretin, allopurinol, atrazine, ondomycin, ipratropium, amifostine, aminoglutarimide, amsacrine, anastrozole, citalopram, arsenic trioxide, asparaginase, aspirin, azacitidine, azetepa, azotomycin, batimastat, benzotelluride, bevacizumab, bicalutamide, bisanffene, bischafide dimesylate, bizelesin, Bleomycin, brequinar, bropirimine, busulfan, cetuximab, capecitabine, carcimetine, carboplatin, carmustine, carbicin, carpricin, celecoxib, chlorambucil, siromulin, cisplatin, cladribine, cinorbendazole mesylate, cyclophosphamide, cytarabine, dacarbazine, actinomycin D, daunorubicin, decitabine, dexamethasone, dezagunine, dezagunine mesylate, diazocine, docetaxel, doxorubicin, droloxifene, drostanolone, duroxamine, edatroloxamine, etisulin, edotoloxamine, enloxacin, enloplatin, fenretinide, ebuloside, etoposide, etopostatin, faloperazole, fazarapine, fenretinide, fluorouracil, fludarabine, fluorouracil, flucitabine, fluoxetine, fostacin, fulvestrant, gemcitabine, hydroxyurea, demethoxydaunorubicin , ifosfamide, sarcosine, interleukin II (IL-2, including recombinant interleukin II or rIL2), interferon alpha-2a, interferon alpha-2b, interferon alpha-1, interferon alpha-alpha3, interferon beta-1a, interferon gamma-1b, methadone, melenstatin acetate, melphalan, mecamyl, mercaptopurine, methotrexate, metoprine, meturedepa, mitindomide, mitocarcin, mitocromin, mitocromin, mitomycin, mitomycin, mitomycin, mitoxantrone, mitotane, mitoxantrone, mycophenolic acid, nelarabine, nocodazole, nokaramycin, olanzapine, oxadiazine, paclitaxel, polyethylene glycol, pomomycin, pentomustine, amide, pipobroman, piposulfan, piroxapine hydrochloride, mithramycin, prumitorone, porphyrin, Porphinomycin, prednimustine, procarbazine, puromycin, pyrofuran, pyroxithromycin, rogacamide, benzoinol, ulinastatin, pyrazole, stavudine, streamicin, spiramycin, spiracin, spirocycline, zocin, sumonifene, temozole, temoxifen, temoporfin, teniposide, tenilolone, tetrolone, timolol, thioguanine, thiotapa, thiazofuran, tirabasamide, topotecan, toremifene,Terliliron, triptorelin, trastuzumab, uracil mustard, vitampin, vilbiprofen, vinblastine, vincristine sulfate, vindesine, vilbiprofen, vorbutin, vilbiprofen, vinorelbine, vornisolidine, vezoloxi, vorozoo, zenaplatin, zenostatin, zoledronic acid, zolimmycin, and combinations thereof. ,

[0178] Examples of suitable anticancer agents include, but are not limited to, Goodman and Gilman, The Pharmacological Basis of Therapeutics, 12th edition, edited by Laurence Brunton, Bruce Chabner, Bjom Knollman, McGraw Hill Professional, 2010.

[0179] In some exemplary embodiments, the pharmaceutical composition comprises a salt (e.g., a monosalt or a disalt) of compound (1) or compound (10) or an analog thereof and at least one other therapeutic agent, wherein the other therapeutic agent comprises an anti-angiogenic agent, for example, bevacizumab. In one embodiment, the anti-angiogenic agent is selected from aflibercept, axitinib, angiostatin, endostatin, 16 kDa prolactin fragment, laminin peptide, fibronectin peptide, tissue metalloproteinase inhibitor (TIMP 1, 2, 3, 4), plasminogen activator inhibitor (PAI-1, -2), tumor necrosis factor alpha, (high dose, in vitro), TGF-β1, interferon (IFN-α, -β, γ), ELR-CXC chemokine, IL-12; SDF-1; MIG; platelet factor 4 (PF-4); IP-10, thrombospondin (TSP), SPARC, 2-methoxyestradiol, proliferator-activated protein kinase, suramin, sorafenib, tirofenib, thalidomide, Cortisone, mitochondria, fumagillin (AGM-1470; TNP-470), tamoxifen, retinoic acid, CM101, dexamethasone, leukemia inhibitor factor (LIF), hedgehog inhibitors and combinations thereof.

[0180] The drug combination can include the first and second therapeutic agents in any desired ratio, provided that a synergistic or synergistic effect still occurs. The synergistic drug combination preferably contains the first and second therapeutic agents in a ratio of about 1:9 to about 9:1. In one embodiment, the synergistic drug combination provides the first and second therapeutic agents in a ratio of about 1:8 to about 8:1, about 1:7 to about 7:1, from about 1:6 to about 6:1, from about 1:5 to about 5:1, from about 1:4 to about 4:1, from about 1:3 to about 3:1, or from about 1:2 to about 2:1. In one embodiment, the synergistic drug combination comprises the first and second therapeutic agents in a ratio of about 1:1.

[0181] In one embodiment, the second therapeutic agent is selected from Allopurinol, Arsenic Trioxide, Azacedin, Bortezomib, Bevacizumab, Capecitabine, Carboplatin, Celecoxib, Chlorambucil, Clavabol, Cytarabine, Dacarbazine, Daunorubicin HCl, Docetaxel, Doxorubicin HCl, Fluorouracil, Gemcitabine HCl, Hydroxyurea, Ifosfamide, Imatinib Mesylate, Ixabepilone, Lenatramide, Megestrol Acetate, Methotrexate, Mitotane, Mitoxantrone HCl, Oxaliplatin, Paclitaxel, Pradoxicil, Roficillin, Sorafenib, Streptozocin, Tamoxifen Citrate, Topotecan HCl, Tretinoin, Vandetanib, Vismodegib, Vorinostat, and combinations thereof.

[0182] 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.

[0183] Table 2: Classification of drugs

[0184]

[0185]

[0186] In some embodiments, the second therapeutic agent includes a drug targeting tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor. In one embodiment, the second therapeutic agent includes a recombinant TRAIL or agonist antibody that activates one or more TRAIL receptors. In one embodiment, the second therapeutic agent includes one or more antibodies or recombinant TRAIL that activate signal transduction by DR4, DR5 or both. In one embodiment, the second therapeutic agent includes one or more of AMG-655, LBY-135, mapatumumab, lexatumumab, Apomab and rhApo2L / TRAIL. In one embodiment, the second therapeutic agent includes a compound selected from camptothecin, 5-FU, capecitabine, cisplatin, doxorubicin, irinotecan, paclitaxel, cisplatin, bortezomib, BH3I-2, rituximab, radiation, triterpenoids, sorafenib, gemcitabine e, HDAC inhibitors, carboplatin, T-101 (a gossypol derivative), ABT-263, ABT-737 and GX-15-070 (obatoclax), vorinostat, cetuximab, panitumumab, bevacizumab, ganituzumab, interferon gamma, sorafenib, XIAP antagonists, Bcl-2 antagonists and Smac mimetics.

[0187] VI. Dosage

[0188] In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 40, 50, 60 or 100 mg to about 2000 mg; about 4, 6, 6 or 10 mg to about 200 mg; or about 0.4, 0.5, 0.6 or 1 mg to about 20 mg, wherein the weight can be based on the compound in its free base form. In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or its analog, or a pharmaceutically acceptable salt thereof, in a dosage range of about 50 mg to about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900 or 2000 mg; from about 5 mg to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 and 200 mg; or 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 compound (1) or compound (10) or its analog, or a pharmaceutically acceptable salt thereof, in a dosage range of about 40 mg to about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900 or 2000 mg; from about 4 mg to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 mg; or from about 0.4 mg to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 110, 120, 130, 140, 150 15, 16, 17, 18, 19 and 20 mg.In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or its analog, or a pharmaceutically acceptable salt thereof, in a dosage range of about 60 mg to about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900 or 2000 mg; about 6 mg to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 1900 or 2000 mg; about 6 mg to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 1900 or 2000 mg; or 200 mg; or about 0.6 mg to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 mg. In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 100 mg to about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900 mg or 2000 mg; or about 10 to about 20, 30, 40, 50, or about 1 mg to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg. In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or its analog, or a pharmaceutically acceptable salt thereof, in a dosage range of about 200 mg to about 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg; about 20 mg to about 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg; or about 2 mg to about 3, 4, 5, 6, 7, 8, 9, 10, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg. 12, 13, 14, 15, 16, 17, 18, 19 or 20 mg, based on the compound in its free base form.In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 400 mg to about 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900 or 2000 mg; about 40 mg to about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 mg; or about 4 mg to about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 mg, based on the free base form of the compound. In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or its analog or its pharmaceutically acceptable salt in a dosage range of about 50 mg to about 60, 70, 80, 90 or 100 mg; about 60 mg to about 70, 80, 90 or 100 mg; from about 70 mg to about 80, 90 or 100 mg, about 80 mg to about 90 or 100 mg; from about 90 mg to about 100 mg; from about 5 mg to about 6, 7, 8, 9 or 10 mg; about 6 mg to about 7, 8, 9 or 10 mg; about 7 mg to about 8, 9 or 10 mg, about 8 mg to about 9 mg or 10 mg; about 9 mg to about 10 mg; from about 0.5 mg to about 0.6, 0.7, 0.8, 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.

[0189] In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 1 mg / kg to about 40 mg / kg; 0.1 mg / kg to about 4 mg / kg; or 0.01 mg / kg to about 0.40 mg / kg. In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or its analog, or a pharmaceutically acceptable salt thereof, at about 1, 2, 3, 4, 5, 6, 7, 8 or 9 mg / kg to about 10, 20, 30 or 40 mg / kg; about 10, 11, 12, 13, 14, 15, 16, 17, 18 or 19 mg / kg to about 20, 30 or 40 mg / kg; about 20, 21, 22, 23, 24, 25, 26, 27, 28 or 29 mg / kg to about 30 or 40 mg / kg; about 30, 31, 32, 33, 34, 35, 36, 37, 38 or 39 mg / kg to about 40 mg / kg; from 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; from about 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8 or 1.9 mg / kg to about 2, 3 or 4 mg / kg; from about 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 mg / kg to about 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.30, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.40, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, 0.50, 0.51, 0.52, 0.53, 0.54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.60, 0.70, 0.71, 0.72, 0.73, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.90, 0.91, 0. about 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28 or 0.29 mg / kg to about 0.30 or 0.40 mg / kg; or about 0.30 mg / kg about 0.30, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38 or 0.39 mg / kg to about 0.40 mg / kg.

[0190] In one embodiment, the pharmaceutical composition comprises compound (1) or compound (10) or an analog thereof, or a pharmaceutically acceptable salt thereof, in a dosage range of about 37.5 mg / m 2 About 1500 mg / m2 ; about 3.75mg / m 2 About 150 mg / m 2 ; or from about 0.4 mg / m 2 About 15 mg / m 2In one embodiment, the pharmaceutical composition comprises a dosage level ranging from about 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180、185、190、195、200、205、210、215、220、225、230、235、240、245、250、255、260、265、270、275、280、285、290、295、300、305、310、315、320、325、330、335、340、345、 350、355、360、365、370、375、380、385、390、395、440、405、405、315、350、350、310、 315,350,350,310,315,320,325,330,315,320,325,330,335,340,345,330,335,340,345,350,335,340,345,340,315,350,350,310,315,350,350,310,315,340, 350,330,315,340, 345,330,335,340,345,350,335,340,345,350,335,340,345, 350、335、340、345、390、335、340、345、390、335、340、350、340、395、340、350、405、 405、405、405、405、405、405、405、405、410、415、420、425、430、435、440、 445、450、455、460、465、470、475、480、485、490、495、500、505、510、515、520、525、 530、535、540、545、550、555、560、565、570、575、580、585、590、595、600、605、610、 615、620、625、630、635、640、645、650、655、660、665、670、675、680、685、690、695、700、705、710、715、720、725、730、735、740、745、750、755、760、765、770、775、780、785、790、795、8800、805、810、815、820、825、830、835、840、845、850、855、860、865、 870、875、880、885、890、895、900、905、910、915、920、925、930、935、940、945、950、 955、960、965、970、975、980、985、990、995、1000、1005、1010、1015、1020、1025、1030、 1035、1040、1045、1050、1055、1060、1065、1070、1075、1080、1085、1090、1095、1100、1105、 1110、1115、1120、1125、1130、1135、1140、1145、1150、1155、1160、1165、1170、1175、1180、 1185、1190、1195、1200、1205、1210、1215、1220、1225、1230、1235、1240、1245、1250、1255、 1260, 1265, 1270, 1275, 1280, 1285, 1290, 1295, 1300, 1305, 1310, 1315, 1320, 1325, 1330, 1335, 1340, 1345, 1350, 1355, 1360, 1365, 1370, 1375, 1380, 1385, 1390, 1395, 1400, 1405, 1410, 1415, 1420, 1425, 1430, 1435, 1440, 1445, 1450, 1455, 1460, 1465, 1470, 1475, 1480, 1485, 1490, 1495mg / m 2 About 1500 mg / m 2; from about 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99、100、101、 102、103、104、105、106、107、108、109、110,111、112、113、114、115、116、117、118、119、120、121、122、123、124、125、126、127、128、129、130、131、132、133、134、135、136、 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149 mg / m 2 About 150 mg / m 2 or about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 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 15 mg / m 2 .

[0191] VII. Dosage Form

[0192] Suitable pharmaceutical compositions that can be used with the methods described herein can be formulated into dosage forms that can be applied to patients. 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 some embodiments, the oral dosage unit is divided into several smaller doses, which are applied to the subject within a predetermined time period to reduce the toxicity of the therapeutic agent being applied. In some embodiments, the oral dosage unit is applied by tablets or capsules, which contain a controlled release formulation that can contain multiple particles, particles, pills, mini tablets or tablets. In one embodiment, the pharmaceutical composition is a parenteral dosage unit. In one embodiment, the pharmaceutical composition is in the form of intravenous (IV), subcutaneous (SC) and intramuscular (M), rectal (PR) and transdermal administration units as parenteral dosage units. In one embodiment, the pharmaceutical composition is a dosage form selected from sterile solutions, suspensions, suppositories, tablets and capsules. In one embodiment, the composition is an oral dosage form selected from tablets, sachets, capsules, lozenges, syrups, liquids, suspensions and elixirs. In one embodiment, the composition is in an oral dosage form selected from the group consisting of tablets, hard shell capsules, soft gelatin capsules, beads, granules, aggregates, powders, gels, solids and semisolids.

[0193] In some embodiments, pharmaceutical compositions in suitable forms for use in the methods described herein include dermatological compositions suitable for topical administration to the skin. For example, dermatological compositions include cosmetic or pharmaceutically acceptable media. Dermatological compositions for topical administration may include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. In some embodiments, conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners, skin enhancers, etc. may be necessary or desirable and may therefore be used. Examples of suitable enhancers include, but are not limited to, ethers such as diethylene glycol monoethyl ether (commercially available) and diethylene glycol monomethyl ether; surfactants such as sodium laurate, sodium lauryl sulfate, cetyltrimethylammonium bromide, benzalkonium chloride, poloxamers (231, 182, 184), Tweens (20, 40, 60, 80) and lecithin (U.S. Pat. No. 4,783, 450); alcohols such as ethanol, propanol, octanol, benzyl alcohol, etc.; polyethylene glycol and its esters such as polyethylene glycol monolaurate; amides and other nitrogen-containing compounds such as urea, dimethylacetamide (DMA), dimethylformamide (DMF), 2-pyrrolidone, 1-methyl-2-pyrrolidone, ethanolamine, diethanolamine and triethanolamine; terpenes; alkanones; and organic acids, particularly citric acid and succinic acid. It is also possible to use and sulfoxides, such as DMSO and C10MSO, but are less preferred.

[0194] In some embodiments, the pharmaceutical composition is in a dosage form selected from a sustained release form, a controlled release form, a delayed release form, and a responsive release form.

[0195] VIII. How to use

[0196] The compositions and methods described herein have utility in treating a number of disease states, including cancer (e.g., colorectal, brain, and glioblastoma). In one embodiment, the compositions and methods described herein are used to treat diseases such as ocular melanoma, alopecia round cell tumor, chondrosarcoma, multifocal disease, diffuse large cell lymphoma, acute lymphocytic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphoma, anorectal cancer, appendix cancer, astrocytoma, and atypical malformation / rhabdoid tumors. In one embodiment, the compositions and methods described herein are used to treat diseases such as basal cell carcinoma, basal cell nevus syndrome, Gorlin-Nevus syndrome, bile duct cancer, bladder cancer, bone cancer, osteosarcoma and malignant fibrous histiocytoma, brain tumors, breast cancer, bronchial tumors, Burkitt's lymphoma, and spinal cord tumors. In one embodiment, the compositions and methods described herein are used to treat such as carcinoid, carcinoid, carcinoma of unknown primary, central nervous system, atypical malformation / rhabditis, meningitis, central nervous system embryonal tumors, central nervous system lymphoma, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloproliferative disease, colon cancer, colorectal cancer, craniopharyngioma and cutaneous T-cell lymphoma (including but not suitable for, Sezary syndrome and fungal disease). In one embodiment, the compositions and methods described herein are used to treat central nervous system embryonal tumors, endometrial cancer, endothelial blastoma, ependymoma, esophageal cancer, Ewing sarcoma tumor family, extracranial germ cell tumors, extrahepatic somatic cell tumors, extrahepatic bile duct cancer and eye cancer. In one embodiment, the compositions and methods described herein are used to treat such as gallbladder cancer, gastric (gastric) cancer, gastrointestinal carcinoid, gastrointestinal stromal tumor (GIST), germ cell tumors, gestational trophoblastic tumors and gliomas. 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 lymphoma, and hypopharyngeal cancer. In one embodiment, the compositions and methods described herein are used to treat drugs such as Kaposi's sarcoma and kidney (renal cell) cancer. In one embodiment, the compositions and methods described herein are used to treat diseases such as Langerhans cell histiocytosis, laryngeal cancer, lip and oral cancer, liver cancer, lung cancer, non-Hodgkin lymphoma, and primary central nervous system lymphoma. In one embodiment, the compositions and methods described herein are used to treat diseases such as Macroglobulinemia (lymphoplasmacytic lymphoma), malignant fibrous histiocytoma of bone and malignant osteosarcoma, neuroblastoma, medullary epithelioma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous neck carcinoma with occult primary, multiple endocrine neoplasms syndrome, oral cancer, multiple myeloma / plasma cell tumor, mycotic isoflavones, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasms, 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 and paranasal sinus cancer, nasopharyngeal carcinoma and neuroblastoma. In one embodiment, the compositions and methods described herein are used to treat diseases such as oral cancer, oral and oral cancer, oropharyngeal cancer, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian cancer, ovarian germ cell tumors, ovarian epithelial cancer and ovarian low malignant potential tumors. In one embodiment, the compositions and methods described herein are used to treat diseases such as pancreatic cancer, papillomatosis, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, intermediately differentiated pineal myeloid tumors, pineocytomas and Supratentorial primitive neuroectodermal tumors, pituitary tumors, 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 colorectal cancer, renal cell (kidney) cancer, renal pelvis and ureter, respiratory cancer of chromosome 15 involving the NUT gene, retinoblastoma and rhabdomyosarcoma. In one embodiment, the compositions and methods described herein are used to treat high-grade prostate cancer. In one embodiment, the compositions and methods described herein are used to treat medium-grade prostate cancer. In one embodiment, the compositions and methods described herein are used to treat low-grade prostate cancer. In one embodiment, the compositions and methods described herein are used to treat castration-resistant prostate cancer.

[0197] The inventors have found that ONC201 (compound (1)) has broad anticancer activity, low toxicity, including low amounts (if any), adverse reactions, low genotoxicity, and high bioavailability, including oral bioavailability, in in vitro and in vivo models, animal models, and human clinical trials. These characteristics allow ONC 201 and various analogs to be particularly suitable for pediatric patients. These characteristics also make ONC 201 and various analogs particularly suitable for chronic treatment, high-risk patients, and to ensure durable responses or stable disease or prevent disease recurrence.

[0198] In one embodiment, the compositions and methods described herein are used to treat pediatric cancers (e.g., pediatric solid tumors, pediatric sarcomas, pediatric Ewing's sarcoma, pediatric gliomas, pediatric central nervous system cancers, pediatric leukemias, and pediatric lymphomas).

[0199] In one embodiment, the compositions and methods described herein are used to treat proliferative skin diseases, such as psoriasis. In one embodiment, the compositions and methods described herein are used to treat cancer selected from salivary gland cancer, sarcoma, Sézary syndrome, skin cancer, eye cancer, skin cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous neck cancer with occult primary and epithelial primitive neuroectodermal tumors. In one embodiment, the compositions and methods described herein are used to treat cancer selected from T-cell lymphoma, testicular cancer, laryngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell cancer of the renal vessels and ureters, and gestational trophoblastic tumors. In one embodiment, the compositions and methods described herein are used to treat cancer selected from cancer of unknown primary, cancer of unknown primary, unusual cancers of children, transitional cell cancer of the renal vessels and ureters, urethral cancer, uterine sarcoma. In one embodiment, the compositions and methods described herein are used to treat cancer selected from vaginal cancer and Vulvar cancer. In one embodiment, the compositions and methods described herein are used to treat cancer selected from Wilms tumor and women's cancer.

[0200] In some embodiments, the compositions and methods described herein are used as first-line treatment (sometimes referred to as primary treatment). In some embodiments, the compositions and methods described herein are used as second-line treatment. In some embodiments, the compositions and methods described herein are used as third-line treatment. In some embodiments, the compositions and methods described herein are used as salvage therapy. The term "salvage therapy" refers to a therapeutic agent that can be taken with any regimen after the subject's initial treatment regimen has failed or after the subject's condition has not responded to the initial treatment. In some embodiments, the compositions and methods described herein are used as rescue treatment. In one embodiment of rescue treatment, the composition is used as a rescue agent to resist the effects of the initial treatment. In one embodiment of rescue treatment, the composition is used as a rescue agent, which is administered to a subject who has developed resistance to standard or initial treatment. In some embodiments, the compositions and methods described herein are used as neoadjuvant therapy. In one embodiment, neoadjuvant therapy includes administering one or more therapeutic agents described herein to a subject before the main line or first line of treatment. In one embodiment, neoadjuvant therapy reduces the size or extent of the cancer being treated before administering the main or first line of treatment to the treated subject. In some embodiments, the compositions and methods described herein are used as adjuvant therapy. In one embodiment, adjuvant therapy comprises administering to a subject one or more therapeutic agents described herein, wherein the one or more therapeutic agents modify the effect of other therapeutic agents already administered to the subject or are administered to the subject simultaneously or subsequently.

[0201] In some embodiments, the compositions and methods described herein show a reduced chance of drug interactions. In some embodiments, Compound (1) or Compound (10) or analogs thereof are eliminated from the patient's body prior to interacting with other pharmaceutically active agents.

[0202] In some embodiments, the compositions and methods described herein exhibit toxicity levels that facilitate combination with other pharmaceutical agents.

[0203] The methods and compositions described herein are not limited to specific animal species. In one embodiment, the subject treated according to the methods and compositions described herein can be a mammal or a non-mammal. In one embodiment, mammal subject mammals 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, cattle, pigs, sheep, goats or rabbits. In one embodiment, non-mammal subjects include, but are not limited to, birds such as ducks, geese, chickens, or turkeys. In one embodiment, the subject is a human. Regardless of sex or 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.

[0204] The methods and compositions described herein are not limited to a particular age of the subject. In one embodiment, the subject treated according to the methods and compositions described herein may be over fifty years old, over fifty-five years old, over sixty years old, over sixty-five years old. In one embodiment, the subject treated according to the methods and compositions described herein may be under fifty years old, under fifty-five years old, under sixty years old, under sixty-five years old.

[0205] In one embodiment, the subject treated according to the methods and compositions described herein can be a pediatric patient. In one embodiment, the pediatric patient is less than 18 years old, less than 17 years old, less than 16 years old, less than 15 years old, less than 14 years old, less than 13 years old, less than 13 years old, less than 12 years old, less than 10 years old, less than 10 years old, less than 9 years old, less than 8 years old, less than 7 years old, less than 6 years old, less than 5 years old, less than 4 years old, less than 3 years old, less than 2 years old, less than 1 year old. In one embodiment, the pediatric patient is less than 12 months old, less than 11 months old, less than 10 years old, less than 9 months old, less than 8 months old, less than 7 months old, less than 6 months old, less than 5 months old, less than 4 months old, less than 3 months old, less than 2 months old, less than 1 month old. In one embodiment, the pediatric patient is less than 4 weeks old, less than 3 weeks old, less than 2 weeks old, less than 1 week old. In one embodiment, the pediatric patient is less than 7 years old, less than 6 years old, less than 5 years old, less than 4 years old, less than 3 years old, less than 2 years old, or less than 1 year old. In one embodiment, the pediatric patient is a neonate. In one embodiment, the pediatric patient is born prematurely. In one embodiment, the pediatric patient is a neonate.

[0206] 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, less than 15 kg weight, less than 14 kg weight, less than 10 kg weight, less than 5 kg weight, less than 4 kg weight, less than 3 kg weight, less than 2 kg weight, or less than 1 kg weight.

[0207] 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, hydroxynocycline, vincristine, prednisone, cytarabine, cisplatin, rituximab, 5-fluorouracil, oxaliplatin, leucovorin, or lenalidomide.

[0208] In one embodiment, the subject has been treated with radiation. In one embodiment, the subject has been treated with surgery. In one embodiment, the subject has received adoptive T cell therapy.

[0209] In some embodiments, the cancer is no longer resistant to ibrutinib, bortezomib, carfilzomib, temozolomide, bevacizumab, cyclophosphamide, hydroxynorapycin, vincristine, prednisone, cytarabine, cisplatin, rituximab, 5-fluorouracil, oxaliplatin, leucovorin, lenalidomide, radiation, surgery, or a combination thereof.

[0210] In some embodiments, the compositions and methods described herein have a dose-response relationship in cancer cells that is different than the dose-response relationship of the same compositions and methods in normal cells. Figure 1 , for example, illustrates the dose-response relationship of compound (1) on normal and tumor cell proliferation and cell death. Figure 1 Cell viability after treatment with compound (1) at the indicated concentrations for 72 hours is shown. Tumors tested included human colon cancer cell line (HCT116), breast tumor cell line (MDA-MB-231) and human primary glioblastoma cell line (U87). Normal cells tested included human foreskin fibroblasts (HFF), human fetal lung fibroblasts (MRC-5) cells and human lung fibroblast cell line (WI-38). Doxorubicin was used as a positive control at 1 μg / mL in normal fibroblasts. Figure 1 As shown, the cell viability of normal cells is at least about 75% at a concentration of about 1-5 mg / mL of compound (1), while the viability of tumor cells is significantly reduced (e.g., equal to or less than 50%) at the same concentration of compound (1). In addition, as the concentration of compound (1) increases beyond about 5 mg / mL, the viability of tumor cells decreases to less than 25%, while the viability of normal cells remains at about 75%.

[0211] Figure 2 The cell viability in human fetal lung fibroblast (MRC-5) cells was measured after 72 hours of treatment with compound (1) (5 μM) or DMSO, indicating a complete recovery period after treatment in drug-free medium. The time point is the time after compound (1) was removed after 72 hours of treatment. Figure 2 As shown, cell recovery was observed with compound (1), but not DMSO.

[0212] In some embodiments, the compositions and methods described herein have practical value in treating cancer. In one embodiment, the compositions and methods described herein can be used to treat cancer in human subjects. In some embodiments, the treatment method comprises administering to a subject in need of such treatment a pharmaceutically effective amount of compound (1) or compound (10) or an analog thereof, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0213] In one embodiment, the method of treatment comprises administering to a subject in need of such treatment: (i) a first therapeutic agent comprising compound (1) or compound (10) or an analog thereof, or a pharmaceutically acceptable salt thereof, and (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 compound (1) include the following dihydrochlorides:

[0214]

[0215] It is understood that the dihydrochloride salt of compound (1) or its analogs (including but not limited to compounds of formula (10)) or alternative di-salts obvious from the teachings of the present disclosure can replace compound (1) or its analogs in the compositions or dosage regimens described herein.

[0216] In some embodiments, the method of treatment comprises administering a synergistic drug combination simultaneously or sequentially, the synergistic drug combination comprising (i) a first therapeutic agent comprising compound (1) or compound (10) or an analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) a second therapeutic agent. In one embodiment, the method of treatment comprises administering a therapeutically synergistically effective amount of a first therapeutic agent and a second therapeutic agent simultaneously or sequentially to a subject in need of such treatment. In one embodiment, the method of treatment comprises administering an effective amount of a first therapeutic agent in combination with an effective amount of a second therapeutic agent, the combination of which provides a synergistic effect in vivo to treat a cancer that is sensitive to the combination, and wherein the first and second therapeutic agents are administered simultaneously or sequentially. In one embodiment, the method of treatment comprises administering to a subject of such treatment, an effective amount of a first therapeutic agent in combination with an effective amount of a second therapeutic agent, wherein the combination provides a synergistic effect in vivo to treat minimal residual disease that is sensitive to the combination, and wherein the first and second therapeutic agents are administered simultaneously or sequentially.

[0217] In some embodiments, the second therapeutic agent can be administered before or prior to the first therapeutic agent.

[0218] In one embodiment, the method of treating is for a cancer selected from a solid tumor, a liquid tumor, a lymphoma, a leukemia, or a myeloma.

[0219] In one embodiment, the method of treatment is directed to a solid tumor, wherein the solid tumor is selected from the group consisting of: cervical cancer, endometrial cancer, extracranial germ cell tumors; extracranial germ cell tumors germ cell tumors; gestational trophoblastic tumors; ovarian cancer, ovarian germ cell tumors, ovarian epithelial cancer and ovarian malignant malignancies; penile cancer, prostate cancer; gestational and breast cancer high-grade prostate cancer; medium-grade prostate cancer; low-grade prostate cancer; castration-resistant prostate cancer; breast cancer; bile duct cancer; extrahepatic bile duct cancer; gallbladder cancer hepatocellular carcinoma (liver cancer) kidney (renal cell) cancer; liver cancer, renal cell (kidney) cancer, renal pelvis and ureter; basal cell carcinoma; basal cell nevus syndrome, Gorlin-Nevus syndrome, melanoma, Merkel cell carcinoma, papillomatosis, multiple endocrine neoplasia syndrome; pancreatic cancer, parathyroid cancer, ocular melanoma; eye cancer; Retinoblastoma; malignant fibrous histiocytoma of the Ewing sarcoma family of tumors; defibrotic round cell tumor; chondrosarcoma, Kaposi's sarcoma, rhabdomyosarcoma; spinal cord tumors, meningitis, CNS embryonal tumors, chordoma, CNS embryonal tumors, endothelioma, ependymoma, neuroblastoma; intermediately differentiated pineal parenchymal tumors, pineocytoma; adrenocortical carcinoma; bone cancer, malignant fibrous histiocytoma of the bone and osteosarcoma; osteosarcoma and malignant fibrous histiocytoma of the bone; carcinoid tumors, carcinoma of unknown primary, bronchial tumors, lung cancer, pleural alveolar tumors; NUT gene involvement on chromosome 15 respiratory cancer, astrocytoma, Atypical teratoma / rhabdoid tumor; Central nervous system atypical teratoma / rhabdoid tumor, craniopharyngioma, glioma, brain cancer, medulloblastoma, myeloepithelioma, epithelial primitive neuroectodermal tumor; pituitary tumors; gastric cancer (stomach), gastrointestinal carcinoid, gastrointestinal stromal tumor (GIST), bladder cancer, anorectal cancer, adnexal cancer, esophageal cancer, hypopharyngeal cancer; laryngeal cancer, lip and oral cavity cancer, metastatic squamous cell carcinoma with occult primary, oral cancer, nasal and paranasal sinus cancer, nasopharyngeal cancer, oral cancer, oral and oral cavity cancer, oropharyngeal cancer, paranasal sinus and nasal cavity cancer, pharyngeal cancer; head and neck cancer and mesothelioma.

[0220] In one embodiment, the method of treating a lymphoma is selected from the group consisting of diffuse large 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, primary central nervous system lymphoma, Hodgkin's lymphoma, Macroglobulinemia; mycoses, fungal primary central nervous system lymphoma; lymphoplasmacytic lymphoma and primary central nervous system lymphoma.

[0221] In one embodiment, the method of treating is directed to a non-Hodgkin lymphoma (NHL) selected from the group consisting of mantle cell lymphoma, diffuse large cell lymphoma, follicular lymphoma, marginal zone lymphoma, small lymphocytic lymphoma, lymphocytic lymphoma NHL, Waldenstrom's macroglobulinemia, and cutaneous lymphoma.

[0222] In one embodiment, the method of treatment is for a leukemia selected from: acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloproliferative disease; hairy cell leukemia acute myeloid leukemia (AML); chronic myeloid leukemia (CML); and Langerhans cell histiocytosis.

[0223] In one embodiment, the method of treating is for an acute leukemia selected from the group consisting of acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, myelodysplastic syndrome, and a myeloproliferative disorder.

[0224] In one embodiment, the method of treating 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; multiple myeloma / plasma cell neoplasm, multiple myeloma, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasms, and myeloproliferative disorders.

[0225] In one embodiment, the method of treating is for a cancer selected from the group consisting of acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphomas, anal or rectal cancer, appendix cancer, astrocytoma, and atypical malformations / neoplasms.

[0226] In one embodiment, the method of treating a cancer is selected from the group consisting of basal cell carcinoma, basal cell nevus syndrome, Gorlin-Nevus syndrome, bile duct cancer, bladder cancer, bone cancer, osteosarcoma and malignant fibrous histiocytoma, brain tumors, bronchial tumors, Burkitt's lymphoma, and spinal cord tumors.

[0227] In one embodiment, the method of treating is for a cancer selected from the group consisting of: carcinoid tumors, carcinoma of unknown primary, CNS atypical malformation / rhabditis, CNS embryonal tumors, CNS lymphomas, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, and cutaneous T-cell lymphoma (including, but not limited to, Sezary syndrome and mycosis fungoides).

[0228] In one embodiment, the method of treating a cancer is selected from the group consisting of embryonal tumors of the central nervous system, endometrial cancer, endothelial blastoma, ependymoma, esophageal cancer, Ewing sarcoma family of tumors, defibrotic round cell tumor, chondrosarcoma, extracranial germ cell tumors, germ cell tumors, extrahepatic bile duct cancer, and eye cancers, including intraocular melanoma and retinoblastoma.

[0229] Choose a cancer option: Gallbladder cancer, stomach cancer (gastric cancer), gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (GIST), germ cell tumors, gestational trophoblastic tumors, and gliomas.

[0230] In one embodiment, the method of treating is for a cancer selected from the group consisting of hairy cell leukemia, head and neck cancer, hepatocellular carcinoma (liver cancer), histiocytosis, Hodgkin lymphoma, and hypopharyngeal cancer.

[0231] In one embodiment, the method of treatment is for a cancer selected from Kaposi's sarcoma and kidney (renal cell) cancer.

[0232] In one embodiment, the method of treating a cancer is for a cancer selected from the group consisting of 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 lymphoma, and primary central nervous system lymphoma.

[0233] In one embodiment, the treatment method targets a cancer selected from: macroglobulinemia (lymphoplasmacytic lymphoma), malignant fibrous histiocytoma of the bone and osteosarcoma, neuroblastoma, medullary epithelioma, melanoma, Merkel cell carcinoma, mesothelioma, occult primary metastatic squamous cell carcinoma, multiple endocrine neoplasms syndrome, oral cancer, multiple myeloma / plasma cell neoplasm, mycoses, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasms, multiple myeloma and myeloproliferative disorders.

[0234] In one embodiment, the method of treatment is useful for treating a cancer selected from the group consisting of: nasal cavity and paranasal sinus cancer, nasopharyngeal carcinoma, and neuroblastoma.

[0235] In one embodiment, the method of treatment can be used to treat a cancer selected from the group consisting of: oral cancer, oral and oral cancer, oropharyngeal cancer, osteosarcoma and malignant fibroblastoma of bone, ovarian cancer, ovarian germ cell tumor, ovarian epithelial cancer, and ovarian low malignant potential tumor.

[0236] In one embodiment, the method of treatment can be used to treat a cancer selected from the group consisting of pancreatic cancer, papilloma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, intermediately differentiated pineal parenchymal tumors, pinewood tumors and epithelial primitive neuroectodermal tumors, pituitary tumors, pleuropulmonary tumors, pregnancy and breast cancer, primary central nervous system lymphoma and prostate cancer.

[0237] In one embodiment, the method of treatment can be used to treat a cancer selected from colorectal cancer, renal cell (kidney) cancer, kidney stones and urinary tract cancers, respiratory tract cancers of chromosome 15 involving the NUT gene, retinoblastoma and rhabdomyosarcoma.

[0238] In one embodiment, the method of treatment can be used to treat a cancer selected from the group consisting of: salivary gland cancer, sarcoma, Sézary syndrome, skin cancer, skin cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, occult primary squamous neck cancer, and supratentorial primitive neuroectodermal tumor.

[0239] In one embodiment, the method of treatment is useful for treating a cancer selected from the group consisting of: T-cell lymphoma, testicular cancer, laryngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell cancer of the renal vascular and ureter, and gestational trophoblastic tumor.

[0240] In one embodiment, the method of treatment is a useful option for treating cancers: cancer of unknown primary site, cancer of unknown primary site, unusual cancers of childhood, transitional cell cancer of the renal vascular and ureter, urethral cancer, and uterine sarcoma.

[0241] In one embodiment, the method of treatment is for treating a cancer selected from the group consisting of: vaginal cancer and carcinoma of the vagina.

[0242] In one embodiment, the method of treatment is for treating a cancer selected from the group consisting of: Wilms tumor and women's cancer.

[0243] In some embodiments, cancer treatment includes preventing tumor growth in a subject with cancer. In some embodiments, cancer treatment includes preventing the formation of cancer metastases. In some embodiments, treatment of cancer includes targeted therapy for minimal residual disease in a subject with cancer known to have minimal residual disease in the cancer or a subject at risk for minimal residual disease.

[0244] This may be indicated after surgery to treat the primary tumor and / or after chemotherapy (radiation therapy) has been started or determined to be effective. Disseminated tumor cells may be dormant and usually cannot be attacked by chemotherapy (radiation therapy). A patient so treated appears to be in a state of healing and is considered to be "minimal residual disease". However, if dormant cells become metastatic tumors due to growth stimulation after a longer dormant state, they have the potential to form metastases.

[0245] As used herein, "minimal residual disease" refers to the few cancer cells remaining in a subject during or after treatment when the subject is in remission (showing no disease symptoms or signs). The methods described herein are preferably applicable to the forms of the diseases listed herein, including the adult and pediatric forms of these diseases.

[0246] In one embodiment, the method of treatment can be used to treat an autoimmune disease. Autoimmune diseases include, but are not limited to, alopecia areata, antiphospholipids, autoimmune liver disease, 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, syndrome, systemic lupus erythematosus and vitiligo.

[0247] In one embodiment, the method of treatment can be used to treat autoimmune and inflammatory diseases of the peripheral nervous system such as amyotrophic lateral sclerosis (Lou Gehrig's disease) which have various causes, such as metabolic disorders including diabetes, B12 and folate vitamin deficiencies, chemotherapy drugs and drugs used to treat HIV, toxins that cause peripheral nerve damage, cancer that develops peripheral neuropathy, paraneoplastic syndromes, alcohol abuse, chronic kidney disease, injuries that cause compression of nerves and other pathologies, such as Lyme disease, Guillain-Barre syndrome, connective tissue diseases, rheumatoid arthritis, Sjögren's syndrome, systemic lupus erythematosus, certain inflammatory conditions such as sarcoidosis, celiac disease, hereditary diseases such as charcotmarie tooth syndrome, Friedreich's ataxia and / or idiopathic diseases where no specific cause is found but inflammatory and / or autoimmune mechanisms are responsible for the onset of the disease.

[0248] In one embodiment, the treatment methods can be used to treat autoimmune and inflammatory diseases with ocular manifestations. Such ocular manifestations include, but are not limited to, ocular cicatricial pemphigoid, Moren's corneal ulcer, various forms of uveitis, rheumatoid arthritis, systemic lupus erythematosus, polyarteritis nodosa, relapsing polychondritis, Wegener's granulomatosis, scleroderma, Behcet's disease, Reiter's disease, inflammatory bowel disease (ulcerative colitis and Crohn's disease) and ankylosing spondylitis, retinitis pigmentosa, macular degeneration, keratoconjunctivitis, scleritis, parascleritis, keratitis, peripheral corneal ulcers, and less common entities such as choroiditis, retinal vasculitis, scleral nodules, retinal detachment and / or macular edema.

[0249] In one embodiment, the method of treatment can be used to treat acute allogeneic transplant rejection in transplant patients. In one embodiment, the method of treatment can be used to treat ischemic stroke. In one embodiment, the method of treatment is useful for treating inflammatory diseases, including but not limited to arthritis, psoriasis, asthma and colitis.

[0250] In one embodiment, the therapeutic agent comprises a pharmaceutically acceptable monosalt of compound (1) or an analog thereof (e.g., a compound of formula (10)). In one embodiment, the therapeutic agent comprises a pharmaceutically acceptable disalt of compound (1) or an analog thereof (e.g., a compound of formula (10)). As described herein, some analogs may be trisalts. In one embodiment, the therapeutic agent comprises compound (1) or an analog thereof (e.g., a compound of formula (10)), in a pharmaceutically acceptable form of a monosalt or disalt selected from hydrochloride, hydrobromide, bisulfate, sulfate, phosphate, fumarate, succinate, oxalate and lactate, bisulfate, hydroxy, tartrate, nitrate, citrate, tartrate, carbonate, malate, maleate, fumarate sulfonate, methanesulfonate, formate, acetate and carboxylate. In one embodiment, the therapeutic agent comprises the chemical compound (1) or its analog in the form of a pharmaceutically acceptable salt selected from p-toluenesulfonate, benzenesulfonate, methanesulfonate, oxalate, succinate, tartrate, citrate, fumarate and maleate. In one embodiment, the therapeutic agent comprises compound (1) or its analog in the form of a pharmaceutically acceptable mono- or di-salt with a counterion selected from ammonium, sodium, potassium, calcium, magnesium, zinc, lithium, and / or with a counterion such as methylamino, dimethylamino, diethylamino, triethylamino counterion and combinations thereof. In one embodiment, the therapeutic agent comprises a di-salt of a compound described herein in the form of a halide, such as a dihydrochloride or dihydrobromide.

[0251] In some embodiments of the method of treatment, the second therapeutic agent comprises an anticancer agent. In some embodiments of the method of treatment, the second therapeutic agent is selected from, but is not limited to, avermectin, aclarubicin, atradazole, aksunin, aspirin, alderleukin, ivermectin, allopurinol, atrazine, cyclomycin, isoproterenol, amifostine, aminoglutarimide, anastrozole, anseramicin, arsenic trioxide, asparaginase, aspergillus, azacitidine, azetepa, azotomycin, batimastat, benzoglucose, bevacizumab, bicalutamide, bisantrene, bisnafidedimesylate, bizelesin, bleomycin, brequinar, bropirimine, busulfan, cetuximab, capecitabine, capecitabine, carboplatin, carmustine, carbinocin, carnectin, Carfilzomib, celecoxib, chlorambucil, siromulin, cisplatin, cladribine, cinorle mesylate, cyclophosphamide, cytarabine, dacarbazine, actinomycin D, daunomycin, decitabine, dexamethasone, deazaguanine, dezagunin mesylate, diazolin, docetaxel, doxorubicin, droloxifene, dibromomorphone, duzolimine, edatroloxamine, etisulin, edatroloxamine, Cisplatin, etoposide, etopostatin, etizole, etoposide, etopostatin, fadomazole, famoxicillin, fenretinide, fluorouracil, fludarabine, fluorouracil, flucitabine, fluoxetine, forsterol, fulvestrant, gemcitabine, hydroxyurea, daunorubicin, ifosfamide, sarcosine, interleukin II (IL-2, including recombinant interleukin II or rIL2), interferon alpha-2a, interferon alpha-2b, interferon alpha-1, interferon alpha-3, interferon beta-1a, midazolam, midazolam, meropenem, mecamyl, mecamyl, mercaptopurine, methotrexate, metropine, meturedepa, mimodime, metropine, metropine, mitomycin, mitomycin, mitomycin, mitomycin, mitosper, mitotane, mitoxantrone, mycophenolic acid, nelarabine, nocodazole, nokaramycin, olanzapine, oxyconazole, paclitaxel, pegaspargase, promoxetine, promoxetine, pipobroman, piposulfan, piroxapine hydrochloride, mithramycin, plethora, porfimromycin, porfimromycin, prednimustine, procarbazine, puromycin, pyrimethamine, riboprine, rogletimide, safingol, semustine, simtrazene, sparsosycin, sparsomycin, spirogermanium, spiromustine, streptozocin, sulofenur, tebimycin, tamoxifen, tegaram, tegafur,Tetostatin, temoporfin, teniposide, teroxirone, testolactone, thiamiprine, thioguanine, thiotepa, thiazofuran, telapasitamide, topotecan, toremifene, trestolone, vitamin B, vitamin B, verinolactone, vitamin B, verbicycline, vezoloxi, vezopolol, zenaplatin, zonotactin, zoledronic acid, zolimcin and combinations thereof. ,

[0252] In some embodiments of the method of treatment, the second therapeutic agent is selected from, but not limited to, hormone analogs and anti-hormones, aromatase inhibitors, LHRH agonists and antagonists, growth factor inhibitors, growth factor antibodies, growth factor receptor antibodies, tyrosine kinase inhibitors; antimetabolites; antitumor antibiotics; platinum derivatives; alkylating agents; antimitotic S; tubuline inhibitors; PARP inhibitors, topoisomerase inhibitors, serine / threonine kinase inhibitors, tyrosine kinase inhibitors, protein-protein interaction inhibitors, MEK inhibitors agents, ERK inhibitors, IGF-1R inhibitors, ErbB receptor inhibitors, rapamycin analogs, amifostine, anagrelide, clodronic acid, filgrastim, interferon, interferon alpha, folinic acid, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate and porphyrin, 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-alethine, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A 105972, A 204197, abiraterone, aldesleukin, acitretin, alloceptin-7, amoxicillin, amoxicillin, anthraquinone, amoxicillin, amoxicillin, amoxicillin, apoxicillin, apoxicillin, apoxicillin, AG-2037, AP-5280, aposide, apoinsulin, arabinose, agliflozin, arzoxifene, ataracetam, atiracetam, aspirin PE, AVLB, AZ10992, ABX-EGF, AMG-479 (ganituzumab) 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, azacytidine, azabetalol B, azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, Diploid gibberellins, BCX-1777, BKM-120, bucomitin, BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW 2992 (afatinib, tomtovok),BIBF 1120 (vargatef), BI 836845, BI 2536, BI 6727, BI 836845, BI 847325, BI 853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, bryostatin-1, bortezomib, myristylcaprolactone, cholethiophene, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, canertinib, canfosfamide, capecitabine, carboxybiphenylplatinum, CCl-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, cefotaxime, ceftriaxone, celecoxib, celmoleukin, cemadotin, CH4987655 / RO-4987655, chlortriphenylamine, cilengitide, cyclosporine, CDA-II, CKD-602, CKI-27, clofarabin, 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, cyanomorpholino doxorubicin, cytarabine, D24851, decitabine, deoxyrubicin, deoxyvitamin, deoxybexycin, deoxycorticosterone B, dexamethasone, decilostat, diethylstilbestrol, diflomotecan, DMD, dolastatin 10, dolastatin, DS-7423, E7010, E-6201, edatrexat, edotreotide, efaproxiral, eflornithine, EGFR inhibitor, EKB-569, EKB-509, enzastaurin, enzalotide, efacin, epothilone B, 86526, erlotinib, ET-18-0CH3, ethynylcytidine, ethinyl estradiol, exatecan mesylate, exemestane, exisulind, fenretinide, etomitumab, floxuridine, folic acid, FOLFOX, FOLFOX4, FOLFIRI, mesultan, flutoxetine, camptothecin, gallium maltolate, gefinitib, GLC-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100, gp100 peptide vaccine, GSK-5126766, GSK-690693, GSK-1120212 (trametinib),GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron, Herceptin, hexamethylmelamine, histamine, homoserine, hyaluronic acid, hydroxyurea, hydroxyprogesterone caproate, ibandronate, itrafenib, itrafenib, itrafenib, IDN-5109, IGF-1R inhibitor, IMC-1C11, IMC-A12 (cixutumumab), immunohistochemistry, hemolysin, interferon alpha-2a, interferon alpha-2b, pegylated interferon alpha-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafamib, ipilimumab, isoflavone, isoflavone B, isoflavones, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, kahalid F, ketoconazole, KW-2170, KW-2450, roplatin, leflunomide, renostelin, leuprolide, rifampin, lesadron, LGD-1550, linezolid, lutetyl tuxaphylline, lomotrexol, loosxantrone, LU 223651, lurtotecan, LY-S6AKT1, LY-2780301, mafosfamide, maramastat, 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, neridronate, neratinib, Nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, norethin, N-acetylcysteine, 6-benzylguanine, ormelin, omeprazole, oncophage, oncoVEXGM-CSF, orplatin, 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, perillylalcohol, pertuzumab, PI3K inhibitor, PI3K / mTOR inhibitor, PG-TXL, PG2, PLX-4032 / -5185426 (vemurafenib), PLX-3603 / RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pivaloyloxymethylbutyrate, pyrone, Phenoxydiol O, PKI166, xplevitrexed, mithramycin, polymalonic acid, ranisol, prednisolone. ,

[0253] In some embodiments of the method of treatment, the second therapeutic agent is selected from tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate, bicalutamide, aminoglutarimide, cyproterone acetate, finasteride, buserelin acetate, fluocortolone, fluoxymesterone, octreotide and combinations thereof. In some embodiments of the method of treatment, the second therapeutic agent is selected from, but not limited to, the group consisting of LHRH agonists and LHRH antagonists. In some embodiments, the LHRH agonist is selected from goherelin acetate, flubiroline acetate, triptorelinpamoate and combinations thereof. In some embodiments, the second therapeutic agent includes an LHRH antagonist selected from Degarelix, Cetrorelix, Abarelix, Ozarelix, a combination of Degarelix. In some embodiments of the method of treatment, the second therapeutic agent includes an inhibitor of a growth factor. In some embodiments, inhibitors of growth factors are selected, but not limited to, from 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), selected from HER2, HER3 and HER4.

[0254] In some embodiments of the method of treatment, the second therapeutic agent comprises a tyrosine kinase inhibitor. In some embodiments of the method of treatment, the tyrosine kinase inhibitor is selected from, but not limited to, cetuximab, gefitinib, imatinib, lapatinib and trastuzumab and combinations thereof. In some embodiments of the method of treatment, the second therapeutic agent comprises an aromatase inhibitor. In some embodiments of the method of treatment, the aromatase inhibitor is selected from anastrozole, letrozole, liarozole, vorozole, exemestane, atamestane and combinations thereof.

[0255] In some embodiments of the method of treatment, the second therapeutic agent comprises an antimetabolite. In some embodiments of the method of treatment, the antimetabolite comprises an antifolate. In some embodiments of the method of treatment, the preservative is selected from methotrexate, raltitrexed, pyrimidine analogs and combinations thereof. In some embodiments of the method of treatment, the antimetabolite is a pyrimidine analog. In some embodiments of the method of treatment, the pyrimidine analog is selected from 5-fluorouracil, capecitabine, gemcitabine and combinations thereof. In some embodiments of the method of treatment, the antimetabolite is a purine analog or an adenosine analog. In some embodiments of the method of treatment, the purine analog or adenosine analog is selected from mercaptopurine, thioguanine, cladribine and pentostatin, cytarabine, fludarabine and combinations thereof. In some embodiments of the method of treatment, the second therapeutic agent comprises an antitumor antibiotic. In some embodiments of the method of treatment, the antitumor drug, doxorubicin, daunorubicin, doxorubicin and demethoxydaunorubicin, scleromycin-C, bleomycin, actinomycin D, mithramycin, streptozocin and combinations thereof. In some embodiments of the method of treatment, the second therapeutic agent comprises a platinum derivative. In some embodiments of the method of treatment, the platinum derivative is selected from cisplatin, oxaliplatin, carboplatin and combinations thereof. In some embodiments of the method of treatment, the second therapeutic agent comprises an alkylating agent. In some embodiments of the method of treatment, the alkylation chemistries select estramustin, meclorethamine, melphalan, chlorambucil, busulphan, dacarbazin, cyclophosphamide, ifosfamide, temozolomide, nitrosoureas and combinations thereof In some embodiments of the method of treatment, the second therapeutic agent comprises a nitrosourea. In some embodiments of the method of treatment, the second therapeutic agent comprises an antimitotic agent, In some embodiments of the method of treatment, the second therapeutic agent comprises an antimitotic agent. In some embodiments of the method of treatment, the antimitotic agent is selected from vinca alkaloids and taxanes. In some embodiments of the method of treatment, paclitaxel is selected from paclitaxel, docetaxel and combinations thereof. In some embodiments of the method of treatment, the vinca alkaloid is selected from vitamins, verinol, verinol, vincristine and combinations thereof. In some embodiments of the method of treatment, the second therapeutic agent comprises a topoisomerase inhibitor. In some embodiments of the method of treatment, the topoisomerase inhibitor is epipodophyllotoxin. In some embodiments of the method of treatment, the topoisomerase inhibitor, which is an epipodophyllotoxin, is selected from etoposide, fenbifol, teniposide, amsacrin, topotecan, irinotecan, mitoxantron, and combinations thereof. In some embodiments, the second therapeutic agent comprises a serine / threonine kinase inhibitor. In some embodiments of the method of treatment, the serine / threonine kinase inhibitor is selected from a PDK 1 inhibitor, a B-Raf inhibitor, an mTOR inhibitor,mTORC1 inhibitors, PI3K inhibitors, dual mTOR / PI3K inhibitors, STK 33 inhibitors, AKT inhibitors, PLK 1 inhibitors, CDK inhibitors, Aurora kinase inhibitors, and combinations thereof. In some embodiments of the method of treatment, the second therapeutic agent comprises a tyrosine kinase inhibitor. In some embodiments of the method of treatment, the second therapeutic agent comprises a PTK2 / FAK inhibitor. In some embodiments of the method of treatment, the second therapeutic agent comprises a protein-protein interaction inhibitor. In some embodiments of the method of treatment, the protein-protein interaction inhibitor is selected from IAP, Mcl-1, MDM2 / MDMX, and combinations thereof. In some embodiments of the method of treatment, the second therapeutic agent comprises a rapamycin analog. In some embodiments, the rapamycin analog is selected from everolimus, sirolimus, ridaforolimus, sirolimus, and combinations thereof. In some embodiments of the methods of treatment, the second therapeutic agent is selected from aripipridine, anagrelide, clodronic acid, filgrastim, interferon, interferon alpha, leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate, and porfimer, and combinations thereof. In some embodiments of the method of treatment, the second therapeutic agent is selected from 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-alethine, 131-1-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A 105972, A204197, abiraterone, aldesleukin, ivermectin, alloceptin-7, atrazine, alvocidib, amonafide, anthrapyrazole, AG-2037, AP-5280, apaziquone, apomine, avermectin, atratriptan, atratriptan, aspirin, aspirin, AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab), ARRY162, ARRY 438162, ARRY-300, ARRY-142886 / AZD-6244 (selumetinib) / AZD-8330, AR-12, AR-42, AS-703988, AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026 (primasertib), avastin, -2014, azacytidine, azabetalicin B, azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576, bevacizumab, BEZ-235, disodium dilactate, BCX-1777,BKM-120, BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BNP-1350, BNP-7787, BIBW2992 (afatinib, tomtoVok), BIBF1120 (vargatef), BI836845, BI 2536, BI 6727, BI 836845, BI 847325, BI 853520, BUB-022, bleomycin acid, bleomycin A, bleomycin B, brivanib, Brystone-1, bortezomib, brostallicin, busulphan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol, canertinib, canfosfamide, capecitabine, carboxybiphenylplatinum, CC1-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime, cefotaxime, ceftriaxone, celecoxib, celmoleukin, cemadotin, CH4987655 / RO-4987655, clotribenzepam, cyclosporine, CDA-II, CDC-394, CKD-602, CKI-27, clofarabin, 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, cyanomorpholino doxorubicin, cytarabine, D24851, decitabine, deoxyrubicin, deoxyvitamin, deoxythymidine, deoxycephalosporin B, dexamethasone, dexamethasone, diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10, doranidazole, DS-7423, E7010, E-6201, edatrexat, edotreotide, efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509, enzastaurin, itraconazole B, efaproxiral, epothilone B, efaproxiral, ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine, ethinyl estradiol, exatecan, exatecan mesylate, exemestane, exisulind, fenretinide, filgretan, filgrolidon, folic acid, FOLFOX, FOLFOX4, FOLFIRI, gefitinib, gemtuzumab, gefitinib, gefitinib,Gefitinib, GFC-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100, gp100 peptide vaccine, GSK-5126766, GSK-690693, GSK-1120212 (trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron, herceptin, hexamethylmelamine, histamine, homoserine protein, hyaluronic acid, hydroxyurea, hydroxyprogesterone a caproate, ibandronic acid, ibrutinib, idempotent, edengestrol, IDN-5109, IGF-1R inhibitor, IMC-1C11, IM C-A12 (cixutumumab), immunoglobulin, hemolysin, interferon alpha-2a, interferon alpha-2b, pegylated interferon alpha-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib, isoproterenol, isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088, conjugated estrogens, kahalid F, ketoconazole, KW-2170, KW-2450, lopaplatin, leflunomide, leuporelin, lexidronam, LGD-1550, linezolid, lutetium tuxaphyrin, lomotrexol, loosxantrone, LU 223651, lurtotecan, LY-S6AKT1, LY-2780301, mafosfamide, marimastat, mechlorethamine, MEK inhibitors, MEK-162, methyltestosterone, methylprednisolone, MEDI-573, MEN-10755, MDX-H210, MDX-447, MDX-1379, MGV, midostaurin, Minodronic acid, mitomycin, mivobulin, MK-2206, MK-0646 (dalotuzumab), MLN518, motexaf in gadolinium, MS-209, MS-275, MX6, neridronate, neratinib, Nexavar, neovastat, nilotinib, nimesulide, nitroglycerin, nolatrexed, norrelin, N-acetylcysteine, 06-benzylguanine, Oblimersen, omeprazole, oncophage,oncoVEXGM-CSF, orplatin, ortataxel, OX44 antibody, OSI-027, OSI-906 (linsitinib), 4-1BB antibody, oxantrazole, estrogen, panitumumab, 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, perillylalol, pertuzumab, PI3K inhibitor, PI3K / mTOR inhibitor, PG-TXL, PG2, PLX-4032 / RO-5185426 (vemurafenib), PLX-3603 / RO-5212054, PT-100, PWT-33597, PX-866, picoplatin, pivaloyloxymethylbutyrate, vebiventrone, phenoxydiol O, PKI166, plevitrexed, mithramycin, polyadrenaline, prednisolone, quinine, Quinupristin, R115777, RAF-265, ramosetron, rapensetron, rapastatin, RDEA-119 / BAY 869766, RDEA-436, rapamycin analogs, receptor tyrosine kinase (RTK) inhibitors, revimid, RG-7167, RG-7304, RG-7421, RG-7321, rojatocin, ritudronic acid, rituximab, rofesbub, rofecoxib, RO-31-7453, RO-5126766, RO-5068760, RPR109881A, naringenone, erythromycin, R-flurbiprofen, RX-0201, S-9788, SAHA, sargramostim, satraplatin, SB408075, Se-015 / Ve-015, SU5416, SU6668, SDX-101, semustin, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib, spirocycline, veratramine, simvastatin hydroxamic acid, cisplatin, T 900607, T 138067, TAK-733, TAS-103, tacedinaline, talaporfin, Tarceva, tariquitar,tasisulam, taxotere, taxoprexin, testosterone, temofifen, testosterone, testosterone propionate, tesmefen, testosterone, tetracycline, tezazepam, thalidomide, citric acid, enalapril, thymine, thymosin, thiazosin, tivafenib, tirapamide, tovacapine, TransMID-107, retinoic acid, trastuzumab, tremelimumab, tretinoin, triacetyluridine, triapine, triciribine, trimetrexate, TLK-286TXD 258, tykerb / tyverb, urocidin, valrubicin, vatalanib, vincristine, vitamins, virulizin, WX-UK1, WX-554, vectibix, xeloda, XELOX, XL-147, XL-228, XL-281, XL-518 / R-7420 / GDC-0973, XL-765, YM-511, YM-598, ZD-4190, ZD-6474, ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZD1839, ZSTK-474, zoledronat, zosuquidar and combinations thereof.

[0256] In some embodiments, the other therapeutic agent comprises a steroid. Steroids include, but are not limited to, dexamethasone, prednisolone, methylprednisolone, prednisone, hydrocortisone, triamcinolone acetonide, betamethasone and corticosteroids. In some embodiments, the other therapeutic agent comprises an antiemetic. dopamine agonists (such as domperidone, olanzapine, haloperidol, haloperidol, chlorpromazine, prochlorperazine, alizapride, atorvastatin, docetaxel, proclopramide, and metoclopramide), NK1 receptor antagonists (such as aspirin and placebo), antihistamines (such as cimetidine, diphenhydramine, diphenhydramine, doxylcycline, medilide, promethazine, hydroxyzine), cannabinoids (such as marijuana, trigonel, and sativex), benzodiazepines (such as midazolam and lorazepam), anticholinergics (such as hyoscine), trimethobenzamide, ginger, emetrol, propofol, mint, thymol, and ferulic acid.

[0257] The pharmaceutical composition can be administered to the subject via any suitable route of administration. In one embodiment, the pharmaceutical composition is administered orally, parenterally, transdermally or transmucosally. In one embodiment, the pharmaceutical composition is administered parenterally. In one embodiment, the pharmaceutical composition is administered to the subject via a parenteral route of administration intravenously (IV), subcutaneously (SC) and intramuscularly (IM). In one embodiment, the pharmaceutical composition is administered to the subject by a route of administration selected from rectal and transdermal. In one embodiment, the pharmaceutical composition is administered to the subject in a dosage form, selecting a sterile solution, a suspension, a suppository, a tablet and a capsule. In one embodiment, the pharmaceutical composition is administered to the subject in an oral dosage form, selected from tablets, pouches, capsules, lozenges, syrups, liquids, suspensions and elixirs. In one embodiment, the pharmaceutical composition is administered to the subject in an oral dosage form, selected from tablets, hard shell capsules, soft gelatin capsules, beads, granules, aggregates, powders, gels, solids and semisolids.

[0258] In some embodiments, the pharmaceutical composition is administered to a subject as a dosage form selected from a sustained release form, a controlled release form, a delayed release form, and a responsive release form.

[0259] In some embodiments, the pharmaceutical composition is administered to the subject once a day. In some embodiments, the pharmaceutical composition is administered to an infrequent dosing regimen (e.g., once a week or less frequently). In some embodiments, the pharmaceutical composition is administered to the subject according to a frequent dosing regimen (e.g., more than once a week). In some embodiments, the pharmaceutical composition is administered to the subject once a week. In some embodiments, the pharmaceutical composition is administered once every four weeks. In some embodiments, the pharmaceutical composition is administered twice a week. In some embodiments, the pharmaceutical composition is administered once every two weeks. In some embodiments, the pharmaceutical composition is administered once every three weeks. In some embodiments, the pharmaceutical composition is administered to the subject once a week, once every two weeks, once every three weeks, once every four weeks, or a combination thereof in a repetitive cycle.

[0260] In one embodiment, the method of treatment comprises administering to a subject in need of such treatment: (i) a first therapeutic agent comprising compound (1), compound (10) or an analog thereof or a pharmaceutically acceptable salt thereof and (ii) a second therapeutic agent, wherein the first therapeutic agent and the second therapeutic agent are administered simultaneously or sequentially; and further comprises testing the expression of endoplasmic reticulum (ER) stress response genes in a biological sample. In some embodiments, the endoplasmic reticulum stress response genes are selected from the group including, but not limited to, C / EBP homologous protein (CHOP), activating transcription factor 3 (ATF3) and CHOP and ATF3. ATF3, activating transcription factor 4 (ATF4) CHOP, IRE1, binding immunoglobulin (BiP), eukaryotic translation initiation factor 2A (eIF2a), endoplasmic reticulum stress response genes), X-box binding protein 1 (XBP1). The biological sample can be a tumor, peripheral blood mononuclear cells or a skin biopsy. The biological sample can be obtained before, during or after administration. In some embodiments, 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 more of ER stress genes. In some embodiments, the method of treatment further comprises adjusting the dose of the first therapeutic agent to achieve induction of about 50% to about 100%, about 100% to about 150%, about 150% to about 200%, about 200% 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% of ER stress genes. In some embodiments, the method of treatment further comprises adjusting the dose of the first therapeutic agent to achieve an induction of about 50% to about 100%, about 100% to about 200%, about 200% to about 300%, about 300%, about 400%, about 400% to about 500%, about 500% to about 600%, or greater than 600% of ER stress genes.

[0261] In one embodiment, the method of treatment comprises administering to a subject in need of such treatment: (i) a first therapeutic agent comprising Compound (1) or Compound (10), or an analog thereof or a pharmaceutically acceptable salt thereof, and (ii) a second therapeutic agent, wherein the first therapeutic agent and the second therapeutic agent are administered simultaneously or sequentially; and further comprises testing a biological sample for expression of proteasome activity. In some embodiments, the proteasome activity may be tryptase-like, trypsin-like and / or caspase-like activity. In some embodiments, the biological sample may be a tumor, a peripheral blood mononuclear cell, or a skin cell. The biological sample may be obtained before, during, or after administration of the drug. In some embodiments, the method of treatment further comprises adjusting the dosage to achieve about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% proteasome activity. In some embodiments, the method of treatment further comprises adjusting the dosage 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% proteasome activity. In some embodiments, the method of treatment further comprises adjusting the dosage 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% proteasome activity.

[0262] In one aspect, provided herein is a method of treatment comprising administering to a subject in need of such treatment a first therapeutic agent comprising a compound of Formula (1) or Formula (10) or an analog thereof, or a pharmaceutically acceptable salt thereof (e.g., a di-salt or tri-salt) and a second therapeutic agent, the method comprising:

[0263] (i) administering a first therapeutic agent to a subject;

[0264] (ii) waiting until a predetermined waiting time after the first therapeutic agent administration time reaches the subject; and / or until the adverse event is resolved or resolved;

[0265] (iii) administering a second therapeutic agent to the subject, wherein the predetermined waiting time is selected to obtain a delayed therapeutic effect of the first therapeutic agent without increasing the risk of possible combined toxic effects of the first and second therapeutic agents. In some embodiments of the method of treatment, the predetermined waiting time is determined based on the clearance rate of the compound of the first therapeutic agent or its metabolite. In some embodiments of the method of treatment, the predetermined waiting time is determined by a quantitative assessment of renal function and renal parameters. In some embodiments of the method of treatment, the predetermined waiting time is determined by a renal function assay, wherein the test is selected from the blood level of the compound of the first therapeutic agent or its metabolite; the clearance rate of the compound of the first therapeutic agent or its metabolite; the 24-hour urine clearance of the compound of the first therapeutic agent or its metabolite.

[0266] In one embodiment of the method of treatment, the predetermined waiting time is substantially equal to the time required for complete elimination of the compound of the first therapeutic agent or its metabolite 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 the chemical substance of the first therapeutic agent or its metabolite to be eliminated from the subject. In one embodiment of the method of treatment, the predetermined waiting time is substantially equal to the time required for the compound of the first therapeutic agent or its metabolite to be eliminated from the subject's liver. In one embodiment of the method of treatment, the predetermined waiting time is substantially equal to the time required for complete elimination of the compound of the first therapeutic agent or its metabolite from the subject. In one embodiment of the method of treatment, the predetermined waiting time is approximately 4 hours. In other embodiments, the waiting time is 1 day. In some embodiments, the waiting time is until the C of the compound of the first therapeutic agent is reduced. max In other embodiments, the waiting time is after most adverse events are resolved or resolved. In one embodiment of the method of treatment, the predetermined waiting time 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 time is about 1-7 days, about 1-6 days, about 1-5 days, about 1-4 days, about 1-3 days or about 1 to 2 days. In one embodiment, the waiting time is up to 3 weeks. Previously considered "treatment time period".

[0267] When the order of administration of the first therapeutic agent is reversed, it may be after the Cmax of the second therapeutic agent (i.e., the first administered drug) has passed. In one embodiment, the first therapeutic agent may be administered until most or substantially all of the first administered drug has been eliminated from the body, or toxic effects of the first administration have resolved or are resolving.

[0268] In some embodiments, the method of treatment further comprises monitoring levels of a compound of the first therapeutic agent or its metabolite in the subject using a pharmacokinetic analysis. In some such embodiments, monitoring levels of a compound of the first therapeutic agent or its metabolite in the subject using a pharmacokinetic analysis comprises using concentrations of the compound of the first therapeutic agent or its metabolite to construct a pharmacokinetic profile of the compound of the first therapeutic agent or its metabolite in samples obtained from the subject at at least two time points suitable for constructing a pharmacokinetic profile. In some embodiments of the method, which comprises monitoring levels of a compound of the first therapeutic agent or its metabolite in the subject using a pharmacokinetic analysis, the sample is sampled or self-sampled from the subject at the point of care or point of use by a point-of-care device or point-of-use device or a matrix suitable for storing the sample prior to quantification in a laboratory. In some embodiments of the method of treatment, each point-of-care device or point-of-use device is capable of quantifying a compound of the first therapeutic agent or its metabolite. In some embodiments of the method, which comprises monitoring levels of a chemical substance of the first therapeutic agent or its metabolite in the subject, one or more samples of the subject are collected from the point-of-care device or point-of-use device by a biopsy point before analysis in a laboratory, or a device for analysis at the point of care or point-of-use device. In some embodiments of the method, a biopsy is performed within a time interval of 3-8 hours after administration of the first therapeutic agent. In some embodiments of the method, after administration of the first therapeutic agent In some embodiments of the method, the biopsy is performed within a time interval of 1-24 hours after the first therapeutic agent is administered to the subject. In some embodiments of the method, the biopsy is performed after a time interval of 2 days after the first therapeutic agent is administered to the subject. In some embodiments of the method, the biopsy is performed within a time interval of 3 days after the first therapeutic agent is administered. In some embodiments of the method, the biopsy is performed after a time interval of 4 days after the first therapeutic agent is administered. In some embodiments of the method, the biopsy is performed within a time interval of 1-7 days after the administration of the first therapeutic agent.

[0269] In some embodiments of the method of treatment, the pharmacokinetic profile comprises a pharmacokinetic parameter of the first therapeutic agent suitable for guiding the subject being treated. In one embodiment of the method of treatment, the subject's blood (whole blood, plasma or serum) ("C max ”) in a range of about 1000 ng / dL to 1500 ng / dL for the treatment time period. In some embodiments, the treatment time period C max Less than 1500 ng / dL, greater than 85 ng / dL. In one embodiment of the method of treatment, after administering to the subject blood (whole blood, plasma or serum) ("C maxThe maximum concentration of the first therapeutic agent in the treatment period is about 1000 ng / mL to 1500 ng / mL. max Less than 1500ng / mL, greater than 85ng / mL.

[0270] In some embodiments, the subject is administered with a dose of 100 mg / kg of ethanol in the blood (whole blood, plasma or serum) (“C max ”) has a maximum concentration of about 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 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, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480 or 1490 ng / dL to about 1500 ng / dL; about 100, 101, 102, 103, 104, 105 dL to about 150 ng / dL; or about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14 or 14.5 ng / dL to about 15 ng / dL.

[0271] In some embodiments, after treatment, the subject's blood (whole blood, plasma or serum) ("C max”) has a maximum concentration of about 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480 or 1490 ng / mL 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, 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.

[0272] In some embodiments, the maximum concentration of the first therapeutic agent in the blood ("whole blood, plasma or serum") ("C max ”) is selected from about 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480 or 1490 ng / dL. In some embodiments, the final concentration of the first therapeutic agent in the blood (whole blood, plasma or serum) ("C max148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 200, 201, 202, 203, 204, 205 max ”), is selected from about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14 or 14.5 ng / dL.

[0273] In one embodiment, the final concentration of blood (whole blood, plasma or serum) ("C max ”) wherein the first therapeutic agent is selected from about 1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480 or 1490 ng / mL. In one embodiment, the final concentration of blood (whole blood, plasma or serum) ("C max 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 200, 201, 202, 203, 204, 205 max”) is selected from about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14 or 14.5 ng / mL.

[0274] In one embodiment, the final concentration of blood (whole blood, plasma or serum) ("C max ”) wherein the first therapeutic agent is selected from about 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515,525,535,545,555,565,575,585,595,605,615,625,635,645,655,665,675,685,695,705,715, 725,735,745,755,765,775,785,795,805,815,825,835,845,855,865,875,885,895,905,915,925, 935, 945, 955, 965, 975, 985, 995, 1005, 1015, 1025, 1035, 1045, 1055, 1065, 1075, 1085, 1095, 1105, 1115, 1125, 1135, 1145, 1155, 1165, 1175, 1185, 1195, 1205, 1215, 1225, 1235, 1245, 1255, 1265, 1275, 1285, 1295, 1305, 1315, 1325, 1335, 1345, 1355, 1365, 1375, 1385, 1395, 1405, 1415, 1425, 1435, 1445, 1455, 1465, 1475, 1485, 1495 or 1500 ng / dL. In some embodiments, the blood (whole blood, plasma or serum) ("C max63, 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 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148 or 149 ng / dL. In some embodiments, the blood (whole blood, plasma or serum) ("C max 5, 12, 12.5, 13, 13.5, 14 or 14.5 ng / dL.

[0275] In one embodiment, the final concentration of blood (whole blood, plasma or serum) ("C max”) wherein the first therapeutic agent is selected from about 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515,525,535,545,555,565,575,585,595,605,615,625,635,645,655,665,675,685,695,705,715, 725,735,745,755,765,775,785,795,805,815,825,835,845,855,865,875,885,895,905,915,925, 935, 945, 955, 965, 975, 985, 995, 1005, 1015, 1025, 1035, 1045, 1055, 1065, 1075, 1085, 1095, 1105, 1115, 1125, 1135, 1145, 1155, 1165, 1175, 1185, 1195, 1205, 1215, 1225, 1235, 1245, 1255, 1265, 1275, In some embodiments, the blood (whole blood, plasma or serum) ("C max53, 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, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148 or 149 ng / mL. In some embodiments, the most recent large dose in the blood (whole blood, plasma or serum) ("C max 5, 14, 15.5, 16, 17.5, 18, 19.5, 20, 21.5, 22.5, 23, 24.5, 25, 26.5, 27.5, 28, 29.5, 30, 31.5, 32.5, 33.5, 34, 35.5, 36, 37.5, 38.5, 39, 40.5, 41.5, 42.5, 43.5, 44.5, 45.5, 46.5, 47.5, 48.5, 49.

[0276] In one embodiment of the method of treatment, the subject's blood (whole blood, plasma or serum) ("C max ”) is about 85 ng / dL to 1500 ng / dL; from about 8.5 ng / dL to 150 ng / dL; or about 0.85 ng / dL to 15 ng / dL. In some embodiments, the final concentration of the first therapeutic agent in blood (whole blood, plasma, or serum) (“C max”) in which the subject selection is about 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475,485,495,505,515,525,535,545,555,565,575,585,595,605,615,625,635,645,655,665,675, 685,695,705,715,725,735,745,755,765,775,785,795,805,815,825,835,845,855,865,875,885, 895, 905, 915, 925, 935, 945, 955, 965, 975, 985, 995, 1005, 1015, 1025, 1035, 1045, 1055, 1065, 1075, or from about 1495 ng / dL to about 1500 ng / dL;From about 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or from about 149 ng / dL to about 150 ng / dL; or 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 from about 14.5 ng / dL to about 15 ng / dL.;

[0277] In one embodiment of the method of treatment, the dosage range is about 85 ng / mL to 1500 ng / mL of blood (whole blood, plasma or serum) ("C max ”); from about 8.5 ng / mL to 150 ng / mL; or from about 0.85 ng / mL to 15 ng / mL. In some embodiments, the final concentration of the first therapeutic agent in blood (whole blood, plasma, or serum) (“C max”) in which the subject selection is about 85,95,105,115,125,135,145,155,165,175,185,195,205,215,225,235,245,255,265, 275,285,295,305,315,325,335,345,355,365,375,385,395,405,415,425,435,445,455,465,475, 485,495,505,515,525,535,545,555,565,575,585,595,605,615,625,635,645,655,665,675,685, 695,705,715,725,735,745,755,765,775,785,795,805,815,825,835,845,855,865,875,885,895, 905, 915, 925, 935, 945, 955, 965, 975, 985, 995, 1005, 1015, 1025, 1035, 1045, 1055, 1065, 1075, 1085, 1095, 1105, 1115, 1125, 1135, 1145, 1155, 1165, 1175, 1185, 1195, 1205, 1215, 1225, 1235, 1245, 1255, 1495 ng / mL to about 1500 ng / mL;About 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97,98,99, 149 ng / mL to about 150 ng / mL; or about 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 110, 1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1201, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1271, 1280, 1290, 1301, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 14110 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.;

[0278] In some embodiments of the method, the total drug exposure over time, measured as the area under the curve ("AUC") of the drug concentration in the subject's blood (whole blood, plasma, or serum) during the time period following drug administration ranges from about 150 ng / mL to about 8000 ng / mL; about 15 ng / mL to about 800 ng / mL; or about 1.5 ng / mL to about 80 ng hr / mL. In some embodiments, the AUC is less than 8000 ng hr / mL and greater than or equal to 150 ng hr / mL. In some embodiments, the AUC is less than 800 ng hr / mL and greater than or equal to 15 ng hr / mL. In some embodiments, the AUC is less than 800 ng hr / mL and greater than or equal to 15 ng hr / mL. In some embodiments, the AUC is less than 80 ng hr / mL and greater than or equal to 1.5 ng hr / mL.

[0279] In some embodiments of the method, total drug exposure over time is about 100 ng / mL to about 8000 ng / mL AUC; about 10 ng hr / mL to about 800 ng hr / mL; or about 1 ng hr / mL to about 80 ng hr / mL. In some embodiments, the total drug exposure over time is about from about 150, 200, 400, 600, 800, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, 4000, 4200, 4400, 4600, 4800, 5000, 5200, 5400, 5600, 5800, 6000, 6200, 6400, 6600, 6800, 7000, 7200, 7400, 7600, or from about 7800 ng hr / mL to about 8000 ng hr / mL AUC. In some embodiments, the total drug exposure over time is from about 15, 20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 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 an AUC of about 800 ng hr / mL. In some embodiments, the total drug exposure over time is from about 1.5, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 76, or 78 ng hr / mL to about 80 ng hr / mL.

[0280] In some embodiments of the method, the total drug exposure over time is about 100 ng / mL to about 8000 ng / mL AUC, from about 10 ng hr / mL to about 800 ng hr / mL; or about 1 ng hr / mL to about 80 ng hr / mL. In some embodiments, the total drug exposure over time is about 150 ng / mL to about 7800, 7600, 700, 7200, 7000, 6800, 6600, 6400, 6200, 6000, 5800, 5600, 5400, 525000, 5400, 4000, 4600, 4200, 4200, 4000, 3200, 3600, 3400, 3200, 3000, 2800, 2600, 3400, 2200 140, 120, 100, 800, 600, 400 or 200 ng / mL and in some embodiments, the total drug exposure over time is from about 15 ng / 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, 3300, 280, 260, 240, 220, 200, 180, 160, 140, 120, 100, 80, 60, 40 or 20 ng hr / mL AUC. In some embodiments, the total drug exposure over time is from about 1.5 ng / 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 some embodiments, the total drug exposure over time is about 100 ng / mL to about 200 ng / mL AUC; from about 10 ng / mL to about 20 ng hr / mL; or about 1 ng hr / mL to about 2 ng hr / mL.

[0281] In some embodiments of the method, the total drug exposure over time is selected from about 100, 150, 200, 400, 600, 800, 1000, 1200, 1400, 1600, 1800, 2000, 2002, 2400, 2600, 3200, 3000, 3200, 3400, 3600, 3800, 4000, 4200, 4600, 400, 4200, 4200, 4200, 4600, 4800, 5000, 5200, 5400, AUC of 5600, 5800, 6000, 6200, 6400, 6600, 6800, 7000, 720000, 7400, 7600, 7800 and 8000 ng / mL. In some embodiments, the total drug exposure over time is 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 ng / mL. In some embodiments of the method, the total drug exposure over time is 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, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, and 80 ng / mL.

[0282] In another aspect, provided herein is a method of treating or using a composition to treat a disease state comprising administering to a subject in need of such treatment a combination of a first therapeutic agent and a second therapeutic agent, the method comprising:

[0283] (i) administering to the subject a first therapeutic agent comprising a compound of formula (1) or formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof;

[0284] (ii) monitoring the level of a compound of the first therapeutic agent or a metabolite thereof in the subject using a pharmacokinetic assay;

[0285] (iii) administering a second therapeutic agent based on the level of the first therapeutic agent. In some embodiments of the method, the monitoring step includes using the concentration of the compound of the first therapeutic agent or its metabolite to construct a pharmacokinetic profile of the compound of the first therapeutic agent or its metabolite. A plurality of samples obtained from the subject at time points suitable for constructing the pharmacokinetic profile. In some embodiments of the method, at least two samples are collected at the point of care or point of use, which are sampled or self-sampled on a point-of-care device or point-of-use device, or are suitable for storing samples for quantification of the compound or its metabolite in a laboratory. In some embodiments of the method, each point-of-care device or point-of-use device is capable of quantifying the compound or its metabolite. In some embodiments of the method, the pharmacokinetic profile includes pharmacokinetic parameters suitable for guiding administration of the compound or its salt to the subject. In some embodiments of the method, the samples include 2-12 samples. In some embodiments of the method, the sample collection time is up to 8 hours, up to 24 hours, up to 48 hours, or up to 72 hours. In some embodiments of the method, the pharmacokinetic parameters include at least one parameter selected from AUC, AUC inf , T max , C max , time above a threshold, steady-state concentration, absorption rate, clearance rate, distribution rate, terminal T-1 / 2 or a parameter extracted from a non-compartmental pharmacokinetic (PK) or partitioned PK analysis, including an inter-compartmental PK analysis based on a physiological model. In some embodiments of the method, the treatment method also includes generating a report including the pharmacokinetic profile of the subject. In some embodiments of the method, the report includes a recommendation for dosing based on the pharmacokinetic profile of the subject. In some embodiments of the method, a dose reduction of compound (1), an analog thereof, or a pharmaceutically acceptable salt thereof is indicated to reduce the risk of toxicity based on one or more pharmacokinetic parameters. In some embodiments of the method, a dose reduction of a compound or its salt is expressed based on time above a threshold, where the threshold is a drug concentration above which toxicity occurs, or AUC, AUC inf , mean residence time (MRT), an index defining the pharmacokinetic profile, a volume of distribution at steady state (Vss), a terminal phase distribution volume (Vz), or a combination of a group of pharmacokinetic variables to fully describe the pharmacokinetic profile. In some embodiments of the method, the dose adjustment of the indicated compound or its salt is based on one or more pharmacokinetic parameters to increase efficacy. In some embodiments of the method, based on AUC, AUC inf, MRT, an index defining a pharmacokinetic profile, a steady-state volume of distribution (Vss), one or more of the volumes of distribution to indicate an increase in the dose of a compound or its salt at the terminal phase (Vz) or a combination of a set of pharmacokinetic variables to fully describe the pharmacokinetic profile. In some embodiments of the method, the dose of the compound or its salt is adjusted to within 5% to 25% of the desired target value. In some embodiments of the method, each sample is applied to a care device or a point-of-use device for determining the concentration of a compound or its metabolite, wherein the point-of-care device or the point-of-use device includes a lateral flow strip having a structure and composition such that one or more samples are applied to the lateral flow strip so that a portion of the drug in the sample is combined with a component of the lateral flow strip so that a detectable signal proportional to the drug concentration in the applied sample is generated. In some embodiments of the method, the sample is suitable for storage of the sample prior to laboratory quantification. In some embodiments of the method, the sample is stored as a dried blood spot. In some embodiments of the method, the drug concentration is measured by ELISA, LC MS MS, LC UV or LCMS. In some embodiments of the method, the pharmacokinetic parameters include at least one of steady-state concentration, absorption and terminal T1 / 2. In some embodiments of the method, at least one sample is whole blood.

[0286] IX. Multimodal Approach to Treatment

[0287] In one aspect, provided herein is a multimodal treatment method, wherein a compound of formula (1) or formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof, is supplemented with another treatment modality to the patient. In one embodiment, the multimodal treatment method administers a pharmaceutical composition comprising a compound of formula (1) or formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof, to a subject with radiation therapy or radiation determined to be ineffective. In one embodiment, the multimodal treatment method administers a pharmaceutical composition comprising a compound of formula (1) or formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof, to a subject with radiation therapy, wherein the pharmaceutical composition comprises a compound of formula (1) or formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof, and radiation therapy is administered simultaneously or sequentially in any order. In one embodiment, the multimodal treatment method administers a pharmaceutical composition comprising a compound of formula (1) or formula (10), or an analog thereof, or a pharmaceutically acceptable salt thereof, to a subject with radiation therapy arranged sequentially. In one embodiment, the multimodal treatment method comprises administering a compound comprising Formula (1) or Formula (10) or its analog or its pharmaceutically acceptable salt thereof to a subject in need of such treatment simultaneously with radiation therapy. In one embodiment, the multimodal treatment method is used to treat cancer. In one embodiment, the multimodal treatment method comprises administering a compound comprising Formula (1) or Formula (10) or its analog or its pharmaceutically acceptable salt thereof to a subject in need of such treatment for cancer, and irradiating the cancer cells with a radiation beam. In one embodiment, the multimodal treatment method uses conformal radiation therapy (CRT) to provide a dose volume histogram (DVH) prescribed to a cancer subject. In one embodiment, the multimodal treatment method uses intensity modulated radiation therapy (IMRT) technology to deliver radiation to cancer cells. In one embodiment, the multimodal treatment method uses a technique to compensate for the motion of a subject's tumor during treatment (e.g., where the radiation dose must be applied to a chest tumor that moves when the patient breathes). For example, the multimodal treatment method uses four-dimensional computed tomography (4D CT) scanning technology to adjust the delivered radiation field to compensate for tumor motion during the respiratory cycle.

[0288] The multimodal treatment method can use any suitable type of radiation, including administration of graded gamma radiation, IMRT (intensity modulated radiation therapy), gamma knife, proton therapy and brachytherapy. Radiation therapy and administration of compounds of formula (1) or formula (10), or analogs thereof or pharmaceutically acceptable salts thereof, can be used to treat brain tumors, such as glioblastoma or diseases that have metastasized to the brain (lung cancer). The multimodal treatment method can be used to treat lung cancer, pancreatic cancer, colorectal cancer, breast cancer, sarcoma, prostate cancer, gynecological malignancies and lymphoma. Gamma knife is often used to treat brain metastases. In one embodiment, the multimodal treatment method includes the use of proton therapy to treat cancer, including brain tumors, prostate cancer and tumors near any important organs. Among them, it is very important to minimize toxicity to nearby normal tissues.

[0289] In one embodiment, the multimodal treatment method comprises administering to a subject with cancer in need of such treatment a pharmaceutical composition comprising a compound of formula (1) or formula (10) or an analog thereof, or a pharmaceutically acceptable salt thereof, simultaneously or in combination with adoptive cell therapy (e.g., CAR-T (JCAR 14, 15, 16, 17, KTE-C19 or CTL019); other T cells (AFM13) or NK (CDNO-109 or NK-92)).

[0290] In one embodiment, the multimodal treatment approach eliminates minimal residual disease without increasing the toxicity resulting from treatment with a compound of formula (1) or formula (10) or an analog thereof or a pharmaceutically acceptable salt thereof. In one embodiment, the multimodal treatment approach improves prognosis and / or reduces adverse side effects associated with the disease state or condition being treated in the subject.

[0291] X. Additional derivatives and chemical substances of compound (1) and related compounds

[0292] In one aspect, provided herein are analogs and related salts of compound (1) and processes for preparing the same. One skilled in the art will appreciate that the same general principles and concepts as described above in conjunction with compounds (1), (10) and their salts, including principles and concepts related to methods and pharmaceutical compositions, have equal efficacy for derivatives, analogs and salts of compound (1) and its salts.

[0293] In one embodiment, the analog has the structure of compound (25):

[0294] Wherein Y represents NR4 or O, and wherein R1, R2, R3 and R4 independently represent hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, thioaralkyl, alkanoyl, sulfhydryl, thioalkyl, thioaryl, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl and heterocyclic free radical. In some embodiments, R1, R2, R3 and R4 are optionally substituted. In some embodiments, some or all of the hydrogen atoms in R1, R2, R3 and R4 can be replaced by deuterium. In other embodiments, the analog has the structure of compound (25), wherein R1, R2, R3 and R4 are independently selected from 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, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine is optionally C 1-4 Alkyl, hydroxy or halogen substituted. In even other embodiments, the analog has the structure of compound (25), wherein R1, R2, R3 and R4 are independently selected from 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.

[0295] In one embodiment, the analog has the structure of compound (26):

[0296] wherein R1 and R2 independently represent hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, thioaralkyl, alkanoyl, thioalkyl, thioaryl, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl and heterocyclic radical. In some embodiments, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine and C 1-4Alkylthienyl, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone and C 1-4 Benzyl-piperazine is optionally C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted. In some embodiments, R1 is selected from 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 some embodiments, R2 is selected from 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.

[0297] In some embodiments, R1 is a benzyl group, optionally substituted with 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 wherein X represents a halogen, including a fluorine, chlorine, bromine or iodine atom, preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine. In some embodiments, R2 is a substituent of the benzyl ring, one or more of the substituents, or a combination of substituents 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 wherein X represents a halogen.

[0298] In some embodiments, R1 is hydrogen. In some embodiments, R1 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl group is replaced by C 1-4Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0299] In some embodiments, R2 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl is C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In some embodiments, the arylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3. In some embodiments, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In some embodiments, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In some embodiments, heterocycloalkylalkyl or heteroarylalkyl is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted. In some embodiments, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0300] In one embodiment, the analog has the structure of compound (27):

[0301] wherein R1 represents hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, thioaralkyl, alkanoyl, sulfhydryl, thioalkyl, thioaryl, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl and heterocyclic radical. In some embodiments, R1 is selected from 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, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone and C 1-4 Benzyl-piperazine is optionally C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4Alkyl or halogen substituted. In some embodiments, R1 is selected from 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.

[0302] In some embodiments, R1 is benzyl, optionally substituted with 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 -OCpX 2p+1 , wherein p is an integer from 2 to 20 and wherein X represents a halogen, including a fluorine, chlorine, bromine or iodine atom, preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine. In some embodiments, R1 is hydrogen. In some embodiments, R1 is a substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0303] In one embodiment, the analog has the structure of compound (28): wherein R1 and R2 independently represent hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, thioaralkyl, alkanoyl, thioalkyl, thioaryl, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl and heterocyclic radical. In some embodiments, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine and C 1-4 Alkylthienyl, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone and C 1-4 Benzyl-piperazine is optionally C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4Alkyl or halogen substituted. In some embodiments, R1 is selected from H, CH3, CH2Ph, CH2-((2-Cl)-Ph), CH2-(2-thienyl), CH2CH2Ph, CH2-(2,4-diF-Ph), CH2-((2-CH3)-Ph), CH2CHOHPh, CH2CH2(4-N-benzyl-piperazine), (CH2)3CO-4F-Ph. In some embodiments, R2 is selected from 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, (CH2)3CO-4F-Ph. In some embodiments, when R1 represents CH2Ph, R2 does not represent CH2-((2-CH3)-Ph.

[0304] In some embodiments, R1 is any substituent of the benzyl ring, one or more of the substituents, or substitutions 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 wherein X represents a halogen, including a fluorine, chlorine, bromine or iodine atom, preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine. In some embodiments, R2 is a substituent of a benzyl ring, one or more of the substituents, or a combination of substituents 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 wherein X represents a halogen.

[0305] In some embodiments, R1 is hydrogen. In some embodiments, R1 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0306] In some embodiments, R2 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl is C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In some embodiments, the arylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3. In some embodiments, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In some embodiments, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In some embodiments, heterocycloalkylalkyl or heteroarylalkyl is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted. In some embodiments, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0307] In one embodiment, the analog has the structure of compound (29):

[0308] wherein R1 and R2 independently represent hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, thioaralkyl, alkanoyl, thiol, thioalkyl, thioaryl, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl and heterocyclic radical. In some embodiments, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine and C 1-4 Alkylthienyl, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone and C 1-4 Benzyl-piperazine is C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4Alkyl or halogen substituted. In some embodiments, R1 is selected from 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 some embodiments, R2 is selected from 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, (CH2)3CO-4F-Ph. In some embodiments, when R1 represents CH2Ph, R2 does not represent CH2-((2-CH3)-Ph.

[0309] In some embodiments, R1 is benzyl, optionally substituted with 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 wherein X represents a halogen, including fluorine, chlorine, bromine or iodine atoms, preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine. In some embodiments, R2 is a substituent of the benzyl ring, one or more of the substituents, or a combination of substituents 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 wherein X represents a halogen.

[0310] In some embodiments, R1 is hydrogen. In some embodiments, R1 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0311] In some embodiments, R2 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl is C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In some embodiments, the arylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3. In some embodiments, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In some embodiments, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In some embodiments, heterocycloalkylalkyl or heteroarylalkyl is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted. In some embodiments, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0312] In one embodiment, the analog has the structure of compound (30):

[0313] wherein R1 and R2 independently represent hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, thioaralkyl, alkanoyl, thiol, thioalkyl, thioaryl, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl and heterocyclic radical. In some embodiments, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine, C 1-4 Alkylthienyl, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine is arbitrarily replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4Alkyl or halogen substituted. In some embodiments, R1 is selected from 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 some embodiments, R2 is selected from 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 some embodiments, when R1 represents CH2Ph, R2 does not represent CH2-((2-CH3)-Ph.

[0314] In some embodiments, R1 is benzyl, optionally substituted with 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 wherein X represents a halogen including fluorine, chlorine, bromine or iodine atoms, preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine. In some embodiments, R2 is a substituent of a benzyl ring, one or more of the substituents, or a combination of substituents 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 wherein X represents a halogen.

[0315] In some embodiments, R1 is hydrogen. In some embodiments, R1 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0316] In some embodiments, R2 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl is C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In some embodiments, the arylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3. In some embodiments, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In some embodiments, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In some embodiments, heterocycloalkylalkyl or heteroarylalkyl is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted. In some embodiments, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0317] In one embodiment, the analog has the structure of compound (31):

[0318] wherein R1 and R2 independently represent hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkyloxy, thioaralkyl, alkanoyl, sulfhydryl, thioalkyl, thioaryl, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl and heterocyclic radical. In some embodiments, R1 and R2 are independently selected from H, C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine, C 1-4 Alkylthienyl, where C 1-4 Alkyl, C 1-4 Alkylphenyl, C 1-4 Alkyl phenyl ketone, C 1-4 Benzyl-piperazine is arbitrarily replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4Alkyl or halogen substituted. In some embodiments, R1 is selected from 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, (CH2)3CO-4F-Ph. In some embodiments, R2 is selected from 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 some embodiments, when R1 represents CH2Ph, R2 does not represent CH2-((2-CH3)-Ph.

[0319] In some embodiments, R1 is benzyl, optionally substituted with 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 wherein X represents a halogen including fluorine, chlorine, bromine or iodine atoms, preferably fluorine, chlorine or bromine, more preferably fluorine or chlorine. In some embodiments, R2 is a substituent of a benzyl ring, one or more of the substituents, or a combination of substituents 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 wherein X represents a halogen.

[0320] In some embodiments, R1 is hydrogen. In some embodiments, R1 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl group is replaced by C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted.

[0321] In some embodiments, R2 is substituted or unsubstituted aralkyl, such as benzyl or phenethyl. In some embodiments, the aralkyl is C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 In some embodiments, the arylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3. In some embodiments, R2 is substituted or unsubstituted heterocycloalkylalkyl, such as morpholinoalkyl or piperazinylalkyl groups. In some embodiments, R2 is substituted or unsubstituted heteroarylalkyl, such as isoxazolidinylmethyl or pyridylmethyl groups. In some embodiments, heterocycloalkylalkyl or heteroarylalkyl is substituted with C 1-4 Alkyl, C 1-4 Alkoxy, hydroxy, perhalogenated C 1-4 Alkyl or halogen substituted. In some embodiments, the heterocycloalkylalkyl or heteroarylalkyl is substituted with one or more substituents selected from the group consisting of halo, -CH3, -CF3 and -OCH3.

[0322] XI. Examples

[0323] It should be understood that the description and specific examples provided below are only for illustrative purposes and are not intended to limit the scope of the present disclosure. The following examples are intended to illustrate disclosed embodiments and should not be construed as limiting thereof. Other compounds except those described below can be prepared using the above-mentioned following reaction scheme or its appropriate variation or modification.

[0324] Example 1. Synthesis of 2-chlorobenzylamino-2-imidazoline hydrochloride

[0325] 2-Chlorobenzylamine (141 mg, 1.0 mMol) was added to a solution of 2-methylthio-2-imidazoline hydrochloride (244 mg, 1.00 mmol) in anhydrous dioxane (2.0 mL). The reaction mixture was stirred at 70°C for 90 minutes under an argon atmosphere. The solution was cooled to room temperature, filtered on a sinter funnel, washed with cold dioxane (2 mL), and dried in vacuo. Compound 4.HI (R2 = 2-chlorobenzyl) (242 mg, 72%) was obtained as a white solid and used without further purification.

[0326] Example 2. Synthesis of 2-chlorobenzylamino-2-imidazoline

[0327] 1.0 N sodium hydroxide (2 mL) was added to a solution of 2-chlorobenzylamino-2-imidazoline hydrochloride (242 mg, 0.72 mmol) in water (3 mL) at 7°C. The reaction mixture was stirred at 7°C under argon for 30 minutes. Thereafter, methyl chlorobenzene chloride (5 mL) was added and the mixture was stirred for another 5 minutes. The reaction mixture was extracted with methyl chloride (2×2.5 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated. The 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).

[0328] Example 3. Synthesis of methyl-1-benzyl 4-oxo-3-piperidinecarboxylate (Compound (6))

[0329] Triethylamine (6 mL) was added to a stirred solution of methyl-1-benzyl 4-oxo-3-piperidinecarboxyl hydrochloride (5.7 g, 20 mMol) in ethyl acetate (50 mL) at 7°C. The reaction mixture was stirred at 7°C under argon for 30 minutes. The reaction mixture was extracted with ethyl acetate (2×50 mL) and washed with water (50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated. The resulting free base residue (5, R1=benzyl) was a viscous oil that was used without further purification. MS (ESI) 248 (M+H)

[0330] Example 4. Synthesis of ONC902 (Compound (14))

[0331] To a solution of 2-chlorobenzylamino-2-imidazoline (150 mg, 0.72 mMol) in methanolic 1-benzyl-4-oxo-3-piperidinecarboxylate (5,1R=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 hours. Afterwards, the reaction mixture was refluxed from 125° C. to 130° C. for 2 hours. The solvent was removed in vacuo, extracted with ethyl acetate (10 mL), washed with saturated sodium bicarbonate solution (2×10 mL) and water (10 mL). The organic layer was dried over anhydrous Na 2 SO 4 , filtered and evaporated. The crude free base was purified by RP HPLC (10%-40% acetonitrile / water) to give ONC902 TFA salt (228 mg, 50% yield) as a white solid MS (ESI) 407 (M+H).

[0332] Various analogs such as ONC903, 904, 905, 906, 912, 210, 211, 212, 213, 214, 217, 218, 219, 220, 221, 222, 223, 224, 225 and 226 were prepared using the same procedure starting from different benzyl amines.

[0333] Example 5. Synthesis of ONC907 (Compound (19))

[0334] To a suspension of sodium hydride (3.5 g, 88 mmol) in anhydrous toluene (50 mL) at 60°C, dimethyl carbonate (4.32 g, 48.0 mMol) was added dropwise at room temperature over 0.5 hours under a nitrogen atmosphere. After adding a few drops of methanol, a solution of 1-tert-butoxycarbonyl-4-piperidone (4.8 g, 24 mMol) dissolved in anhydrous toluene (20 mL) was added dropwise to the reaction mixture while stirring at 80°C for more than 1 hour. The reaction mixture was stirred at the same temperature for 3 hours, then cooled to 0°C (ice bath) and adjusted to pH 6-6.5 with acetic acid. The resulting cold mixture was diluted with water (10 mL) and adjusted to pH 8 with 5% sodium hydroxide solution. The toluene layer was separated and the aqueous layer was extracted with toluene (20 mL). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The compound was dried in vacuo to give methyl-1-tert-butoxycarbonyl-4-oxo-3-piperidinecarboxylate (5.0 g, 80%). The obtained compound was used for the next reaction without further purification.

[0335] 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 hours. After that, the reaction mixture was refluxed at 125°C to 130°C for 2 hours. The solvent was removed in vacuo, extracted with ethyl acetate (10 mL), 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 cleaved with 10% trifluoroacetic acid in dichloromethane and purified by RPHPLC (10%-40% acetonitrile / water) to give a white solid. MS (ESI) ESI (ESI)) 297 (M+H).

[0336] Example 6. Synthesis of ONC909 (Compound (21))

[0337] A mixture of ONC907 (100 mg, 0.2 mMol), phenyl bromide (55.0 mg, 0.28 mMol) and potassium carbonate (150 mg, 1.0 mMol) in N,N-dimethylformamide (3 mL) was heated to 70°C for 12 hours. The solvent was removed in vacuo and extracted with ethyl acetate (10 mL) and washed with water (5 mL). The organic layer was dried over anhydrous Na2SO4, filtered and evaporated. The crude free base was purified by RPHPLC (10%-40% acetonitrile / water) to give ONC909 (62 mg, 50%) as a white solid, MS (ESI) 401 (M+H) TFA salt.

[0338] The same procedure was used starting from different halides to obtain ONC 910 and 214. Compounds 227, 228, 229 and 230 (plus the compounds listed in Table X in the preparation) were prepared using analogous procedures to Examples 1 and 5 with different benzyl amines.

[0339] Compound ONC911 was prepared from ONC910 by treatment with TFA.

[0340] Compound (72) was prepared by reacting a precursor NH compound prepared similarly to Example 5 with styrene oxide.

[0341] Example 7. Synthesis of ONC908 (Compound (20))

[0342] To a solution of methyl 1-methyl 4-oxo-3-piperidinecarboxylate (185.0 mg, 1.0 mMol) of 2-methylbenzyl-2-amino-2-imidazoline (190.0 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 hours. Afterwards, the reaction mixture was refluxed from 125°C to 130°C for 2 hours. The solvent was removed in vacuo, extracted with ethyl acetate (10 mL), 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 HPLC 10%-40% acetonitrile and water to give ONC908 (270.0 mg, 50%) TFA salt as a white solid MS (ESI) 311 (M+H).

[0343] Example 8. Synthesis of ONC201 (Compound (1))

[0344] Compound (3) (239.7 g, 0.845 mol, 1.6 eq) was added portionwise to 800 mL of saturated NaHCO3 in a 2 L round bottom flask under stirring. n-Butanol (500 mL) was added to the resulting mixture and the mixture was stirred for 30 minutes 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, N2 inlet, thermocouple, condenser and Dean-Stark trap. Compound (5) (100 g, 0.528 mol, 1 eq) and pyridinium p-toluenesulfonate (PPTS) (6.63 gm 0.026 mol, 5 mol%) were added to the flask. The resulting mixture was heated to reflux for 6 hours. Water in the reaction mixture was separated into a Dean-Stark trap as needed. The reflux temperature was increased from 93°C to 118°C. The progress of the reaction was monitored by HPLC. When the peak area of ​​compound (1) on HPLC remains constant over the reaction time, the reaction is stopped.

[0345] Example 9. Synthesis of the di-salt of ONC201 (compound (1)·2HCl)

[0346] Without isolating compound (1), the reaction mixture of Example 8 was washed with 500 mL of water and diluted with methyl tert-butyl ether (MTBE) (800 mL). The organic phase was washed with water (500 mL×2) and transferred to a 3 L three-necked round bottom flask equipped with mechanical stirring, N2Cl, thermocouple, condenser and Dean-Stark separator. While stirring the reaction mixture, 1N HCl in 1N dioxane-METBE solution (4N HCl in dioxane: 300 mL, 1.2 mol, 2.27 equivalents; MTBE: 1200 mL) was added dropwise until no solid HCl precipitated from the reaction mixture during the addition. The reaction mixture was heated to reflux at 60-65° C. for 2 hours. Water was separated into Dean-Stark traps as needed. After cooling to room temperature, the solid precipitate was filtered through a sintered glass funnel and washed with n-butanol-MTBE (1:2, 600 mL) and MTBE (600 mL), respectively. The solid was dried in a vacuum oven at 65°C overnight (16 hours) to give 200 g of a yellow solid.

[0347] The above solid (200 g) was added to a 2L three-necked round-bottom flask equipped with mechanical stirring, N2 inlet, thermocouple and condenser, followed by ethanol (1000 mL). The mixture was heated to reflux at 78°C for 2 hours. After cooling to room temperature, the solid was filtered through a sintered glass funnel and washed with ethanol (200 mL×3). The wet solid was dried in a vacuum oven at 85°C for 3 days until the residual solvent reached the specified value. 120 g of compound (2) was obtained as a white solid with a yield of 49% and a HPLC purity of 99.7%.

[0348] Example 10. Activity of analogs of compound (1)

[0349] Analogs of various exemplary compounds (1) were prepared based on the synthesis described herein. For each of these compounds, the viability of human cancer cells 72 hours after treatment with the compound was measured. The change in potency (relative to ONC201) was determined as shown in the table below.

[0350] Relative potency of analogs of compound (1)

[0351] No. Identification Code R1 R2 Relative potency* 1 ONC201 CH2Ph CH2-((2-CH3)-Ph) N / A 14 ONC902 CH2Ph CH2(2-Cl-Ph) B 15 ONC903 CH2Ph CH2-(2-Thienyl) C 16 ONC904 CH2Ph CH2CH2Ph B 17 ONC905 CH2Ph CH2CH2(4-N-Benzyl-piperazine) C 18 ONC906 CH2Ph CH2-(2,4-diF-Ph) A 19 ONC907 H CH2-((2-CH3)-Ph) C 20 ONC908 CH3 CH2-((2-CH3)-Ph) B 21 ONC909 CH2CH2Ph CH2-((2-CH3)-Ph) B 32 ONC910 (CH2)3-NH-BOC CH2-((2-CH3)-Ph) B 33 ONC911 (CH2)3-NH2 CH2-((2-CH3)-Ph) B 41 ONC210 <![CDATA[CH2Ph]]> <![CDATA[CH2-(3,5-diF-Ph)]]> A 51 ONC211 <![CDATA[CH2Ph]]> <![CDATA[CH2-(3,4-dichloro-phenyl)]]> A 52 ONC212 <![CDATA[CH2Ph]]> <![CDATA[CH2-(4-CF3-Ph)]]> A 53 ONC213 <![CDATA[CH2Ph]]> <![CDATA[CH2-(3,4-diF-Ph)]]> A 54 ONC214 <![CDATA[CD2C6D5]]> <![CDATA[CH2-((2-CH3)-Ph)]]> B 43 ONC217 <![CDATA[CH2Ph]]> <![CDATA[CH2(2-F-Ph)]]> B 55 ONC218 <![CDATA[CH2Ph]]> <![CDATA[CH2(2-CH3,4-F-Ph)]]> A 56 ONC219 <![CDATA[CH2Ph]]> <![CDATA[CH2-(2,4-dichloro-phenyl)]]> A 57 ONC220 <![CDATA[CH2Ph]]> <![CDATA[CH2-((4-OCH3)-Ph)]]> A 34 <![CDATA[CH2Ph]]> <![CDATA[CH2-(3-pyridyl)]]> 35 <![CDATA[CH2Ph]]> <![CDATA[CH2-(3-isoxazolidinyl)]]> B 36 <![CDATA[CH2Ph]]> <![CDATA[CH2CH2-(4-morpholinyl)]]> A 38 H <![CDATA[CH2-(4-CF3-Ph)]]> A 72 <![CDATA[CH2Ph]]> <![CDATA[CH2-(2-F,4-CF3-Ph)]]> A

[0352] *Relative to the potency of ONC201; A indicates a >2-fold increase in the potency of ONC201; B indicates a potency within 2-fold of ONC201; and C indicates a >2-fold decrease in the potency of ONC201.

[0353] In addition, a single dose of compound (52) administered orally or intraperitoneally to mice bearing human colon cancer xenografts resulted in a significant reduction in tumor volume compared to vehicle-treated controls. Compound (52) has a broad therapeutic window, as it is well tolerated in mice up to at least 225 mg / kg.

[0354] Example 11. Dosage regimen

[0355] Compound (1) was administered to tumor-bearing mice using a 7-day cycle according to the following dosing schedule.

[0356] 1) Day 1: 200 mg / kg orally;

[0357] 2) Day 1 / Day 4: 100 mg / kg per dose orally;

[0358] 3) Day 1 / Day 2: 100 mg / kg per dose, orally;

[0359] 4) Day 1: 2 doses of 100 mg / kg orally, each dose separated by 6 hours.

[0360] The effects of dosing regimens were evaluated and compared.

[0361] Example 12. Determination of the target of compound (1)

[0362] Determination of the target spectrum of compound (1) in the human colon cancer cell line HCT116.

[0363] Briefly, compound (33) (ONC911) was immobilized to agarose beads at different immobilization densities. For analysis by quantitative mass spectrometry, HCT116 human colon cancer cells were grown in culture medium with different forms of isotope-labeled amino acids (SILAC = stable isotope labeling by amino acids in cell culture). The corresponding proteomes can be distinguished by the introduced mass differences. Binding experiments were repeated with partial switching of the label to exclude labeling artifacts. Bound proteins were completely eluted from the affinity matrix, separated by SDS-PAGE and trypsin digested. The recovered peptides were analyzed by LC-MS / MS on a LTQ OrbitrapVelos mass spectrometer (Thermo Fisher). The raw data generated by LC-MS / MS were processed by MaxQuant to obtain quantitative protein abundance data.

[0364] By incubation with compound (1), protein enrichment and competition for binding proteins compared to control matrix were analyzed. This binding and displacement pattern is expected for specific target proteins.

[0365] result

[0366] First, HCT116 cells were cultured and metabolically labeled with SILAC. Efficient SILAC encoding was achieved, with the incorporation of isotopic variants of arginine and lysine exceeding 95%. Sufficient cells were prepared for subsequent experiments. Cell extracts were generated by detergent-mediated cell lysis. Additionally, the remaining nuclei were extracted by lysis in the presence of 400 mM NaCl to include nuclear proteins. Cytosolic and nuclear extracts were combined.

[0367] Linker compound (33) (ONC911) was immobilized on agarose beads via its amino groups. Beads with four different immobilization densities of 6 mM, 3 mM, 1 mM and 0.3 mM were prepared. These matrices were used to enrich proteins in HCT116 extracts and to study the displacement of binding proteins by 50 μM compound (1).

[0368] A total of For all coupling densities and replicates, specific enrichment of proteins was observed for the immobilized chemistry (33) (ONC911).

[0369] The number of target candidates increased with increasing immobilization density. Table 3 summarizes the target candidates for compound (1). The highest coupling density (6 mM) enriched by affinity groups and consistent displacement by two replicates of compound (1) were observed among 14 proteins. At a coupling density of 3 mM, two potential target candidates were identified, both shared at high coupling density. At lower coupling densities (1 and 0.3 mM), two and one proteins, respectively, consistently appeared as targets.

[0370] Additionally, several proteins showed enrichment by affinity matrix and displacement by compound (1), but only one was observed to be displaced in two replicates per coupling density. These proteins are referred to as "OK (with outliers)" in Table 3).

[0371] In conclusion, the immobilized compound (33) (ONC911) appears to be functional and is able to specifically enrich proteins from cell lysates. Furthermore, a differential competition with compound (1) at 50 μM was observed.

[0372] Table 3: Overview of identified target candidates

[0373]

[0374]

[0375] Uniprot ID: best Uniprot identifier; Protein Name: protein name according to Uniprot.

[0376] Target classification: Each protein was evaluated with the indicated coupling density for compound ONC216. "OK" indicates that the corresponding protein was consistently enriched and competed for more than 2 independent replicates. "OK (with outliers)" indicates enrichment by affinity matrix and shift for compound (1), but shift was observed only in one of the two replicates.

[0377] Example 13. GPCR antagonism of compound (1)

[0378] ONC201 was evaluated in whole cells, and a functional assay for β-arrestin G protein-coupled receptor (GPCR) activity was performed to directly measure dopamine receptor activity by detecting the interaction of β-arrestin with an activated GPCR that can serve as a reporter. For each dopamine receptor (DRD1, DRD2S, DRD2L, DRD3, DRD4, and DRD5), cell lines overexpressing reporter constructs were expanded from frozen stocks. Cells were seeded into white-walled 384-well microplates in a total volume of 20 μL and incubated at 37°C prior to assay. With antagonist and then agonist challenge at EC80 concentrations. Intermediate dilutions of the sample pool were performed to generate 5× samples in assay buffer. 3.5 μL of 5× sample was added to cells and incubated at 37°C or room temperature for 30 minutes. Vehicle concentration was 1%. 5 μL of 6× EC80 agonist was added to cells in assay buffer and incubated at 37°C or room temperature for 90 or 180 minutes prior to measurement readout. % Antagonist was calculated using the following formula: % Antagonism = 100% x (1 - (average RLU of test sample x average RLU of vehicle control) / (average RLU of EC80 control - average RLU of vehicle control).

[0379] Example 14. Evaluation of the interaction of compound (1) with ions and transporters

[0380] The ability of ONC201 to interfere with the activity of transporter proteins is evaluated to determine the co-administration schedule of ONC201 with transporter substrates. The timing or dose level of ONC201 in combination with other therapeutic agents may be modified based on the results of these tests. Transporters include OATP1B1, OATP1B3, OAT1, OAT3, OCT1, OCT2, MATE1, and MATE2-f solute carriers (SLCs).

[0381] The ability of ONC201 to interfere with efflux proteins was evaluated to determine whether ONC201 could inhibit the ability of these proteins to efflux small molecule substrates. Inhibition of these efflux proteins could increase the efficacy of efflux protein substrates by increasing their intracellular concentrations or altering their biodistribution by binding to ONC201. Efflux proteins include MDR1 and BCRP.

[0382] The interaction of ONC201 with human MDR1 and BCRP transporters was studied in vitro using MDR1 and BCRP overexpressing Madin-Darby canine kidney (MDCKII-MDR1 and MDCKII-BCRP) and parental cells (MDCKII). Bidirectional permeability of each probe substrate in MDCKII-MDR1 and -BCRP and MDCKII monolayers was performed to investigate whether ONC201 is an inhibitor of MDR1 and BCRP. Digoxin and prazosin were used as probe substrates for MDR1 and BCRP, respectively.

[0383] The inhibition test results are summarized in Table 4 below. ONC201 is an inhibitor of MDR1 and BCRP at 200 micromolar. The combination of ONC201 with a MDR1 or BCRP substrate may increase the efficacy of the substrate by increasing the intracellular concentration of the substrate or altering its biodistribution.

[0384] Table 4:

[0385]

[0386] AB: apical to basolateral; BA: basolateral to apical; PC: positive control

[0387] Example 15. Evaluation of the Potential of Compound (1) as an Inhibitor of P450 Enzymes

[0388] The potential of ONC201 to induce human cytochrome P450 (CYP) enzymes was evaluated, with attention paid to the three major inducible drug-metabolizing enzymes, CYP 1A2, 2B6, and 3A4, using cryopreserved plateable human hepatocytes.

[0389] The experimental CYP induction results of ONC201 are summarized in Table 5 below. ONC201 did not induce P450 in this assay to greater than 20% of the positive control. Therefore, it can be used in combination with other drugs without altering the activity of CYP enzymes.

[0390] Table 5: Induction of CYP mRNA in artificial hepatocytes with various treatments

[0391]

[0392] Calculated using the standard ΔΔCT method a CYP mRNA fold induction values, with 18S gene as reference gene and vehicle control as baseline treated hepatocyte target (CYP) gene expression.

[0393] ΔΔC T Methods The 18S gene was used as a reference gene and the vector control was used as a benchmark to treat the expression of hepatocyte target (CYP) genes.

[0394] b NC: Negative control - Flumaxetine (25 μM) was used as a negative control treatment

[0395] c PC: Positive Control - Omeprazole (50 μM), phenobarbital (750 μM) and rifampicin (25 μM) were used as positive control treatments for CYP1A2, 2B6 and 3A4, respectively.

[0396] Data were calculated from triplicates.

[0397] ONC201 is an inhibitor of seven (7) human cytochrome P450s (CYPs), namely CYP1A2, 2B6, 2C8, 2C9, 2C19, 2D6 and 3A4, in vitro using eight (8) CYP isoform-specific marker substrate reactions in pooled human liver microsomes (HLM). CYP2C8-mediated N-deethylation of amodiaquine, CYP2C9-mediated 4' hydroxylation of diclofenac, CYP2C19-mediated 4' hydroxylation of S-mephenytoin, CYP2D6-mediated 1' hydroxylation of furan, CYP3A4-mediated 1' hydroxylation of midazolam, and testosterone 6β-hydroxylation.

[0398] ONC201 inhibits CYP isoenzymes (CYP 1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 3A4), IC 50 The values ​​ranged from 34.9 to 428.6 μM (4 to 48 times higher than 9 μM and 40-480 times higher than the average plasma concentration of 0.9 uM at 24 hours), and the inhibitory effect was not significantly time-dependent (see Table 6). These results suggest that ONC201 can be taken with most other drugs without safety concerns associated with drug-drug interactions.

[0399] Table 6:

[0400]

[0401] The expression of ONC201 was determined by fitting the normalized data to a sigmoidal inhibitory nonlinear regression model using GRAPHPAD PRISM. a IC 50 value..

[0402] In the measured concentration range (1.5-500 μM), b >500 - no inhibition >50%.

[0403] Example 16 Clinical trial of oral ONC201 in patients with refractory solid tumors.

[0404] ONC201 induces apoptosis in tumor cells, but not normal cells, at doses that cause high levels of cell death in human cancer cells. The safety profile of ONC201 in GLP safety studies in rats and dogs is consistent with the preferential cytotoxicity of ONC201 in tumors over normal cells in vitro. Thus, the in vitro and in vivo characterization of ONC201 suggests that a broad therapeutic window is highly desirable for cancer therapeutics. A schedule of every 21 days was selected for clinical studies based on preclinical results that demonstrated sustained PD in tumors and after preliminary pilot experiments indicated that more frequent dosing did not appear to significantly increase efficacy in vivo.

[0405] Based on the efficacy and safety of ONC201, as well as the novel involvement of signaling pathways important to many cancers, the clinical introduction of ONC201 in patients with advanced cancer is being conducted. The primary objectives of this first-in-human, phase 1 dose-escalation study are to determine the recommended phase II dose (RP2D) of ONC201 orally administered in patients with advanced cancer, as well as to evaluate the safety and tolerability of the drug. Secondary objectives include pharmacokinetics, pharmacodynamic evaluations, and preliminary anti-tumor activity of ONC201.

[0406] Briefly, this Phase I open-label study treated 10 patients with dose escalation in histologically confirmed advanced solid tumors. An additional 10 patients were accrued in an expansion phase to increase safety experience. Patients received ONC201 orally every 3 weeks at doses ranging from 125 to 625 mg in an accelerated titration design.

[0407] RP2D is defined as 625 mg to achieve 1.5-7.5 mg / mL ( ) max . Saturation reached a plasma concentration of 375 mg, indicating that dose escalation above 625 mg was unnecessary. No grade >1 drug-related adverse events occurred. During the dose escalation phase, the mean number of cycles (21 days) was 3.1. The safety of ONC201 at the RP2D was confirmed in 10 patients in the expansion phase. PK analysis showed a half-life of 9.6 hours and an AUC of 25h.μg / L. Prolonged induction of serum caspase-cleaved keratin 18 and induction of TRAIL were observed. Eight of the 10 patients had stable disease, and one patient with prostate adenocarcinoma experienced prolonged stable disease and continued on study for 27 weeks. One additional patient with endometrial carcinoma had a mixed response.

[0408] ONC201 was well tolerated, had a PK profile with absorbable micromolar plasma concentrations, and showed signs of clinical activity when administered orally at 625 mg every 3 weeks.

[0409] Patients and methods

[0410] ethics

[0411] The study was conducted at Robert Wood Johnson University Hospital / Rutgers Cancer Institute of New Jersey (CINJ) in accordance with the Declaration of Helsinki and the International Conference to Harmonize Good Clinical Practice guidelines and was approved by the relevant regulatory committees and the Institutional Review Board of CINJ. Patients provided written informed consent to participate in the study.

[0412] patient population

[0413] Patients aged 18 years or older with advanced solid tumors who had no or no standard therapy, an ECOG performance status ≥1, and disease assessed by RECIST 1.1 criteria were eligible. If patients received radiation therapy, they had to have a measurable lesion outside the irradiated field. Patients had to have completed all prior cytotoxic chemotherapy for at least 4 weeks, alkyl therapy for at least 6 weeks, molecular targeted agents for at least 28 days, and radiation therapy for at least 14 days before the first dose. All previous treatment-related adverse events, except alopecia and neuropathy, grade ≤ 2, had to be resolved. Patients were required to have normal bone marrow and organ function as defined by the following parameters: absolute neutrophil count ≥1,500 / mcL; platelets ≥1 million / mcL; hemoglobin ≥9.0 mg / dL without transfusion in the previous 2 weeks; total bilirubin within normal range (serum bilirubin ≤1.5×ULN for patients with liver metastases); AST(SGOT) / ALT(SGPT) ≤2.5×ULN; and measured or estimated creatinine clearance ≥40 mL / min / 1.73 m2 with creatinine levels above normal. Exclusion criteria included symptomatic brain metastases or asymptomatic brain metastases treated with steroids, prior bevacizumab therapy, allergic reactions to compounds similar to ONC201, uncontrolled concurrent illness, HIV co-retroviral therapy, cardiac disease, cardiac dysfunction, stroke or seizures, impairment of GI function that could alter ONC201 absorption, pregnancy, and use of hematopoietic colony-stimulating growth factor therapy within 2 weeks prior to initiation of treatment.

[0414] Study design and toxicity assessment

[0415] The design is an open-label, dose-escalation phase I trial of a single dose of ONC201 in patients with advanced refractory tumors, who have exhausted or refused standard treatment options for each indication. Capsules (125 mg) of ONC201 were provided by Oncoceutics Inc (Philadelphia, PA). ONC201 was administered orally once every 21-day cycle using an accelerated dose-escalation design. The oral starting dose was 125 mg (10% no-observed adverse event level in rats and dogs). The study was conducted with a single-patient accelerated dose-escalation design, which was designed to stop if any patient experienced at least a grade 2 adverse event that was potentially related to ONC201. In this case, a traditional 3+3 dose-escalation design will be used. Dose escalation can be performed after the previously dosed cohort completes a treatment cycle and meets the criteria for the next dose level. Registration for each subsequent dose level requires all patients registered at the previous dose level to complete cycle 1 dosing and be evaluated 21 days later to assess safety. Dosage levels ranged from 125 mg to 250 mg, 375 mg, 500 mg, and finally to 625 mg.

[0416] After the RP2D was determined, a 22-patient expansion phase was initiated to enroll additional patients at the RP2D to increase the robustness of the safety data generated by the trial.

[0417] All toxicities were assessed according to the Common Terminology Criteria for Adverse Events version 4. DLTs were defined as drug-related adverse events or abnormal laboratory results occurring during the first cycle of treatment that met any of the following criteria: ≥ Grade 3 non-hematologic toxicity; ≥ Grade 3 nausea, vomiting, or diarrhea that persisted for >72 hours despite optimal antiemetic or antidiarrheal therapy; Grade 3-4 AST / ALT combined with Grade 2 elevation in bilirubin; Grade 4 neutropenia lasting =7 days; Grade 4 neutropenia with fever >38.5°C; Grade 3 neutropenia >Grade 3 infection; Thrombocytopenia of any grade associated with clinically significant bleeding; Grade 4 thrombocytopenia; or Grade 4 anemia that was assessed as not related to disease, disease progression, prevalent illness, or concomitant medications; and was determined by the investigator to be “possibly related,” “probably related,” or “definitely related” to the administration of ONC201.

[0418] Safety evaluation

[0419] Safety evaluations including complete blood counts, serum chemistries, and toxicity were assessed at baseline, then during the first 2 cycles, and then every 3 weeks. Electrocardiogram monitoring was performed before ONC201 administration, then at 15 minutes, 1 hour, and 2 hours after dosing. Adverse events were graded using CTCAE version 4.0. Tumor response was assessed using RECIST every 2 cycles.

[0420] Pharmacokinetic analysis

[0421] Plasma samples for PK were collected at baseline, 30 minutes, 2 hours, 4 hours, 6 hours, 24 hours, 48 ​​hours, and 168 hours after the first dose of ONC201 and before the dose during 2-6 weeks of treatment. PK was analyzed by LC-MS / MS using a validated GLP method to detect ONC201 in human plasma. Version 6.3 ( St. Louis, Missouri) for PK analysis.

[0422] Statistical analysis

[0423] Descriptive statistics were used to analyze safety and tumor response data.

[0424] Pharmacodynamic analysis

[0425] PD blood samples were collected 6 hours after ONC201 treatment cycle 1, on days 2, 3, 8, and 15, and on the day of drug administration for cycles 2 and 3. Serum levels of cleaved cytokeratin 18 (cCK18) were assessed using the M30 test, and total cytokeratin 18 (CK18) were assessed using the M65 test (Perviva AB, Sweden). Other tumor-specific markers were also assessed according to standard of care.

[0426] Tumor type in vitro sensitivity

[0427] The in vitro activity of ONC201 was evaluated in 1020 gene-annotated cell lines from the Cancer Drug Sensitivity Gene Set (http: / / www.cancerrxgene.org). IC50 values ​​were determined 72 hours after treatment by cell viability assay as described previously. For each tumor type, the estimated IC50 values ​​of multiple cell lines were averaged. Tumor types were divided into three different groups, representing tertiles of the mean IC50 values. These groups are referred to as "high", "low" and "medium" in Table 11 based on their tertile classification in the ONC201 sensitivity spectrum.

[0428] result

[0429] Patient characteristics

[0430] In the dose escalation phase, 10 evaluable patients were enrolled in this study. Patient characteristics are listed in Table 7. After the completion of the dose escalation phase, an additional 10 patients were enrolled in the ongoing expansion phase (Table 8).

[0431] Table 7. Demographics and safety experience of ONC201 patients every 3 weeks during the dose escalation phase

[0432]

[0433] Table 8. Patient Demographics and Safety Experience in the ONC201 RP2D Expansion Phase (625 mg every 3 weeks)

[0434]

[0435]

[0436] *Indicates that the patient is still studying.

[0437] Dose escalation approach, determination of RP2D and safety

[0438] The dose cohorts are listed in Table 9. 625 mg was the highest dose achieved and was determined to be the RP2D. The only adverse event in the dose escalation phase that could be attributed to ONC201 was low-grade fever in one patient. One patient enrolled in the highest dose cohort was replaced due to rapid disease progression in cycle 1.

[0439] The only adverse event among the 10 patients in the expansion phase that could be attributed to ONC201 was vomiting in one patient. Both adverse events were grade 1 and reversed rapidly. Laboratory tests and physical examinations revealed no drug-related abnormalities. Similarly, cardiovascular evaluations showed no drug-related effects.

[0440] Table 9: ONC201 dose escalation and expansion cohorts given every 3 weeks

[0441]

[0442] Pharmacokinetics

[0443] Plasma collected at serial time points was used to analyze systemic exposure levels of ONC201 in patients ( Figure 5A and Figure 5B PK parameters were determined for all patients and summarized for the top dose cohort (Table 10). Although dose escalation involved a single patient cohort, the PK parameters were compared using AUC and C max The measured systemic exposure of ONC201 appeared to be saturated at a dose of 375 mg ( Fig. 6A and Figure 6B ). For the highest dose cohort, the mean C maxThe mean AUC was 26.3 (SD 10.8) hg / mL and the mean CL / F was 27.19 (SD 10.95) L / h. 1 / 2 It was 9.62 (SD 1.76) hours.

[0444] Table 10. Mean ONC201 pharmacokinetic parameters determined in patients receiving 625 mg ONC201 every three weeks.

[0445]

[0446] In general, CL / F was observed to be variable but consistent across all dose groups. No clear relationship was observed between drug CL / F and patient sex and age. Mild trends were observed with patient weight and BSA. An overall increase in CL / F was observed as weight and BSA increased. Although a slight upward trend was observed, CL / F and CL CR There is no strong correlation between them.

[0447] Stronger correlations were observed between volume of distribution estimates (VZ) and patient weight and BSA. A significant increase in VZ was observed with increasing patient weight. From this trend, VZ was predicted to be greater than 2-fold with an increase in body weight from 50 to 100 kg. Similar trends were observed between VZ and BSA. The effect of dose-normalized exposure parameters on patient weight was further explored. max A trend toward decreasing exposure with increasing body weight was observed in the plots of / dose and AUC / dose versus patient weight. Weight-normalized CL / F plotted versus dose showed a similar trend to the unnormalized CL / F, but with significantly lower variability among patients in the 625 mg dose group.

[0448] Patient response

[0449] Table 11 lists the patient outcomes for the 10 evaluable patients enrolled in the dose escalation phase. Of the 10 evaluable patients, 4 completed at least 4 cycles, and 1 patient received 8 cycles and remains on treatment. On average, patients received 3.1 doses of ONC201. Of the 10 patients in the expansion phase, 6 are still on treatment.

[0450] Table 11. Clinical response and pharmacodynamics during the dose escalation phase.

[0451]

[0452]

[0453] *MR - mixed response, SD stable disease, PD progressive disease. In vitro sensitivity classification by tumor type is described in the Methods section.

[0454] Clinical and laboratory results indicate that the drug is biologically active in the patients treated. Patient #3, a 72-year-old patient with advanced clear cell endometrial cancer had a mixed response with multiple nodes decreasing by >30% with the development of new nodes. Patient #4, a 62-year-old male with renal cancer and bone metastases who had debilitating pain in his clavicle, had relief of pain in his clavicle. Patient #6, a 69-year-old patient with prostate adenocarcinoma experienced prolonged stable disease and is on study for 27 weeks. Patient #8, a 71-year-old patient with colon cancer, had stable disease for 12 weeks with 4 doses of ONC201.

[0455] Pharmacodynamics

[0456] Given the heterogeneity of tumor types in enrolled patients, there are no widely used biomarkers that can be used to uniformly assay all patient samples. In particular, the serum M30 assay is able to detect the caspase-cleaved form of cytokeratin-18 that occurs during apoptosis, which is useful in heterogeneous studies of solid tumors, as most solid tumors express cytokeratin-18. The M30 sandwich ELISA has been widely used in clinical trials as a biomarker of cell death induced by different cancer chemotherapeutic agents in a spectrum of different solid tumors. In addition to the serum M30 assay, the M65 sandwich ELISA assay, which is also used in clinical studies to detect increases in total cytokeratin 18 that may occur with tumor necrosis and disease progression, is used to distinguish tumor cell apoptosis from necrosis.

[0457] As expected, patients with rapid clinical progression who were on study for one cycle showed increases in the M65, but not in the M30 assay. In contrast, patients who remained on study through eight cycles showed increases in the M30, but not in the M65 assay. The four patients who participated in the dose escalation phase of the study progressed to induction in the M30 assay after a single dose of ONC201, typically on day 21 after treatment ( Fig. 7A and Figure 7B To further understand the relevance of the observed heterogeneous M30 induction, the in vitro sensitivities of tumor types defined in a large panel of cell lines from the Gene Set of Drug Sensitivity in Cancer (GDSC) were compared with those of patients in this study ( Figure 8 Interestingly, these patients who underwent induction in M30 were also the 3 patients with tumor types that had high in vitro sensitivity to ONC201 (Table 11).

[0458] Given the downstream induction of TRAIL by ONC201 in preclinical models, serum TRAIL levels were also quantified using an ELISA assay. Half of the patients had moderate ( ) increased, which mostly peaked in the first 24 hours of drug administration.

[0459] discuss

[0460] This example is the first study in humans to investigate the cancer treatment ONC201. The primary objective of the study was to determine the RP2D of oral ONC201 every 3 weeks for patients with solid tumors who have exhausted all treatment options. As expected from the benign preclinical safety of ONC201, no drug-related >1 grade toxicity was observed at micromolar concentrations in preclinical models in any patient. Due to the excellent safety of the drug, the study allowed progression to the next dose level without the need for additional patient enrollment and completed the discreteness of the unaccelerated titration design. The study determined 625 mg, once every 3 weeks, as the RP2D, without toxicity, and this dose achieved therapeutic plasma concentrations. This RP2D exceeds the saturation threshold observed at 375 mg, so there is no need to adjust body surface area to achieve a consistently achievable target blood level. The RP2D has been confirmed in the expansion phase, and an additional 10 patients can be safely evaluated.

[0461] The pharmacokinetic profile of ONC201 demonstrated that the orally administered drug was significantly and rapidly absorbed as shown by the 1.8 hour mean. Importantly, in the highest dose group treated at the RP2D, PK parameters such as Cmax and AUC exceeded those associated with the NOAEL in the GLP toxicology study. The observation of systemic exposure of ONC201 that saturated at 2 dose levels below the RP2D suggests absorption saturation. As absorption saturation occurs at doses that produce apparently well-tolerated therapeutic plasma concentrations, this may serve as a safety feature. These observations support the decision to stop further dose escalation of ONC201 beyond the RP2D while providing a safety margin around the target dose.

[0462] Given that the primary endpoint of the study was based on clinical safety in a highly heterogeneous group of patients with aggressive cancers, it is noteworthy that some patients showed some evidence of clinical benefit. These included a patient with treatment-resistant clear cell endometrial carcinoma with mixed responses, 2 patients who had relief of symptoms related to the site of tumor manifestation, and 2 patients with stable disease for >2 months (prostate and colon adenocarcinoma). In this clinical trial, treatment was terminated after disease progression using RECIST criteria, which specified a 20% increase in tumor size. Signs of anti-tumor activity and the absence of any meaningful side effects in this trial suggest that ONC201 can provide clinical benefit without inflicting the typical toxicities that anti-cancer treatments impose on patients.

[0463] Similar to preclinical findings, PD measurements using the M30 assay showed that the effects of ONC201 were sustained in several patients. Serum TRAIL induction was observed in 2 patients; however, this assay is limited to detecting serum soluble TRAIL because biopsies were not available. The ONC201 PK profile and its sustained PD effect provide an opportunity for staggered dosing of the combination regimen to minimize the risk of drug-drug interactions while maintaining synergistic biological activity. Synergy between ONC201 and approved cancer therapies has been established using taxanes, bevacizumab, bortezomib, and sorafenib.

[0464] In summary, we have shown that ONC201 was well tolerated at the RP2D of 625 mg and demonstrated signs of biological activity in patients with advanced solid tumors.

[0465] Those skilled in the art will appreciate that the exemplary embodiments shown and described above may be changed without departing from the broad inventive concept thereof. Therefore, it should be understood that the present invention is not limited to the exemplary embodiments shown and illustrated, but is intended to cover modifications within the spirit and scope of the present invention as defined by the claims. For example, specific features of the exemplary embodiments may or may not be part of the claimed invention, and features of the disclosed embodiments may be combined. Unless otherwise specified herein, the terms "a", "an" and "the" are not limited to one element, but should be considered as "at least one".

[0466] It should be understood that at least some of the drawings and descriptions of the present invention have been simplified to focus on elements relevant to a clear understanding of the present invention, and for the sake of clarity, other elements of ordinary skill in the art will be understood by those skilled in the art may also comprise a part of the present invention. However, because these elements are well known in the art and because they do not necessarily contribute to a better understanding of the present invention, a description of these elements is not provided here.

[0467] Furthermore, to the extent that the method does not rely on the order of the specific steps set forth herein, the specific order of the steps should not be construed as a limitation on the claims. Claims relating to the methods of the present invention should not be limited to performing their steps in the order written, and those skilled in the art can readily understand that the steps can be varied and still remain within the spirit and scope of the present invention.

[0468] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

Claims

1. A disalt of a compound having the following structure 2. The compound of claim 1, wherein the disalt is selected from the group consisting of hydrochloride, hydrobromide, bisulfate, sulfate, phosphate, fumarate, succinate, oxalate, lactate, tartrate, nitrate, citrate, bitartrate, carbonate, malate, maleate, sulfonate, methanesulfonate, and carboxylate.

3. The compound of claim 1, wherein the disalt is a formate or acetate salt.

4. The compound according to claim 1, wherein the disalt is selected from the group consisting of p-toluenesulfonate, benzenesulfonate, citrate, methanesulfonate, oxalate, succinate, tartrate, fumarate and maleate.

5. The compound according to claim 2, wherein the di-salt is a dihydrochloride or a dihydrobromide.

6. The compound according to claim 2, wherein the di-salt is a dihydrochloride salt.

7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 and a pharmaceutically acceptable carrier.

8. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating cancer.

9. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 7 in the preparation of a medicament for treating a disease selected from the following: desmoplastic round cell tumor, diffuse large B-cell lymphoma, acute lymphocytic leukemia, adrenocortical carcinoma, AIDS-related cancers, anal cancer, rectal cancer, appendix cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, malignant fibrous histiocytoma, breast cancer, bronchial tumors, Burkitt's lymphoma, spinal cord tumors, carcinoids, embryonal tumors of the central nervous system, lymphomas of the central nervous system, cervical cancer, chordoma, chronic lymphocytic leukemia, colon cancer, craniopharyngioma, cutaneous T-cell lymphoma, Sezary syndrome, endometrial cancer, endothelial blastoma, ependymoma, esophageal cancer, Ewing's sarcoma family of tumors , extracranial germ cell tumors, eye cancer, gallbladder cancer, gastric cancer, gastrointestinal stromal tumors, gestational trophoblastic tumors, gliomas, hairy cell leukemia, head and neck cancer, histiocytosis, Hodgkin lymphoma, Kaposi sarcoma, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lung cancer, Waldenstrom's macroglobulinemia, medulloblastoma, medullary epithelioma, melanoma, Merkel cell carcinoma, mesothelioma, plasma cell tumor, nasal cavity cancer, paranasal sinus cancer, neuroblastoma, ovarian cancer, pancreatic cancer, parathyroid cancer, penile cancer, pharyngeal cancer, intermediately differentiated pineal myeloid tumor, pineocytoma, supratentorial primitive neuroectodermal tumor, pituitary tumor, pleuropulmonary blastoma, prostate cancer, renal pelvic cancer, ureteral cancer, cancers involving the NUT gene on chromosome 15, retinoblastoma, and rhabdomyosarcoma.

10. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating a disease selected from the following: renal cell carcinoma, hepatocellular carcinoma, gastrointestinal carcinoid, respiratory tract cancer, osteosarcoma, acute myeloid leukemia, chronic myeloid leukemia or brain tumor.

11. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 7 in the preparation of a medicament for treating a disease selected from the group consisting of germ cell tumors, malignant fibrous histiocytoma of bone and osteosarcoma, myelodysplastic syndrome, astrocytoma, basal cell nevus syndrome, extrahepatic bile duct carcinoma or non-Hodgkin's lymphoma.

12. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 7 in the preparation of a medicament for treating a disease selected from the group consisting of chondrosarcoma, multiple myeloma, atypical teratoid / rhabdoid tumor, Gorlin-nevus syndrome or metastatic squamous cell carcinoma of an occult primary.

13. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating malignant fibrous histiocytoma of bone.

14. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 7 for the preparation of a medicament for treating a disease selected from the group consisting of AIDS-related lymphoma, colorectal cancer, ovarian germ cell tumors, ocular melanoma, myeloproliferative diseases and oral cancer.

15. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating a disease selected from the group consisting of myeloproliferative neoplasms and ovarian tumors of low malignant potential.

16. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating pancreatic cancer.

17. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating primary central nervous system lymphoma, atypical malformation / rhabditis of the central nervous system, epithelial ovarian cancer, hypopharyngeal cancer, nasopharyngeal cancer or oropharyngeal cancer.

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