U.S. patent application number 17/388551 was filed with the patent office on 2022-03-10 for g protein-coupled receptor (gpcr) modulation by imipridones.
The applicant listed for this patent is CHIMERIX, INC.. Invention is credited to Joshua E. ALLEN, Varun Vijay PRABHU, Martin STOGNIEW.
Application Number | 20220072001 17/388551 |
Document ID | / |
Family ID | |
Filed Date | 2022-03-10 |
United States Patent
Application |
20220072001 |
Kind Code |
A1 |
ALLEN; Joshua E. ; et
al. |
March 10, 2022 |
G PROTEIN-COUPLED RECEPTOR (GPCR) MODULATION BY IMIPRIDONES
Abstract
Imipridones has been found to selectively modulate Class A G
protein-coupled receptors (GPCRs), such as the D2-like subfamily of
dopamine receptors, and to be useful for the treatment of
conditions and disorders in need of such modulation, such as
cancers, psychiatric disorders, and bacterial infections. In
addition, methods of identifying whether a subject having these
condition, is likely to be responsive to a treatment regimen, such
as administration of an imipridone, are provided. Furthermore,
methods of assessing the effectiveness of a treatment regimen, such
as administration of an imipridone, monitoring, or providing a
prognosis for a subject with these condition are also provided.
Inventors: |
ALLEN; Joshua E.; (New
Haven, CT) ; STOGNIEW; Martin; (Lakewood Rance,
FL) ; PRABHU; Varun Vijay; (Philadelphia,
PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CHIMERIX, INC. |
Durham |
NC |
US |
|
|
Appl. No.: |
17/388551 |
Filed: |
July 29, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16073228 |
Jul 26, 2018 |
11116771 |
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PCT/US2017/015608 |
Jan 30, 2017 |
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17388551 |
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PCT/US2016/015817 |
Jan 29, 2016 |
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16073228 |
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62425403 |
Nov 22, 2016 |
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62308325 |
Mar 15, 2016 |
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International
Class: |
A61K 31/519 20060101
A61K031/519; A61P 31/04 20060101 A61P031/04; A61P 35/00 20060101
A61P035/00; C07D 471/14 20060101 C07D471/14 |
Claims
1.-142. (canceled)
143. A method for treating a central nervous system cancer in a
subject in need thereof, comprising administering a compound of
formula (1) ##STR00034## or a pharmaceutically acceptable salt
thereof, wherein the central nervous system cancer is susceptible
to modulation by one or more of a Dopamine Receptor (DR) and a G
Protein Coupled Receptor (GPCR).
144. The method of claim 143, wherein the cancer is meningioma,
ependymoma, glioma, neuroblastoma or diffuse intrinsic pontine
glioma.
145. The method of claim 143, wherein the compound is an antagonist
of one or more of the receptors.
146. The method of claim 143, wherein the subject is a human.
147. The method of claim 143, wherein the pharmaceutically
acceptable salt is a di-hydrochloride salt.
148. A method for treating a central nervous system cancer in a
subject in need thereof, comprising administering a compound of
formula (1) ##STR00035## or a pharmaceutically acceptable salt
thereof, wherein the central nervous system cancer has a histone H3
mutation.
149. The method of claim 148, wherein the cancer is meningioma,
ependymoma, glioma, neuroblastoma or diffuse intrinsic pontine
glioma.
150. The method of claim 148, wherein the cancer has an
epigentically silenced unmethylated O(6)-methylguanine-DNA
methyltransferase (MGMT) gene.
151. The method of claim 148, wherein the subject is a human.
152. The method of claim 148, wherein the pharmaceutically
acceptable salt is a di-hydrochloride salt.
153. A method for treating a central nervous system cancer in a
subject in need thereof, comprising administering a compound of
formula (1) ##STR00036## or a pharmaceutically acceptable salt
thereof, wherein the central nervous system cancer has a
epigentically silenced unmethylated O(6)-methylguanine-DNA
methyltransferase (MGMT) gene.
154. The method of claim 153, wherein the cancer is meningioma,
ependymoma, glioma, neuroblastoma or diffuse intrinsic pontine
glioma.
155. The method of claim 153, wherein the cancer has a histone H3
mutation.
156. The method of claim 153, wherein the subject is a human.
157. The method of claim 153, wherein the pharmaceutically
acceptable salt is a di-hydrochloride salt.
Description
[0001] The Sequence Listing associated with this application is
provided in text format in lieu of paper copy, and is hereby
incorporated by reference into the specification. The name of the
text file containing the Sequence Listing is
"81877-339578_2021-11-05_Sequence_Listing_ST25." The text file is
279 KB, was created on Nov. 5, 2021, and is being submitted
electronically via EFS-Web.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0002] This application is a continuation of U.S. application Ser.
No. 16/073,228, filed Jul. 26, 2018, which is a U.S. national stage
application filed under 35 U.S.C. .sctn. 371 from International
Application No. PCT/US2017/015608, filed on Jan. 30, 2017, which is
a continuation-in-part of International Application No.
PCT/US2016/015817, filed on Jan. 29, 2016. International
Application Serial No. PCT/US2017/015608 claims the benefit of U.S.
Provisional Application Ser. No. 62/308,325, filed on Mar. 15,
2016, and U.S. Provisional Application Ser. No. 62/425,403, filed
Nov. 22, 2016, which applications are incorporated by reference as
if reproduced herein and made a part hereof in their entirety, and
the benefit of priority of each of which is claimed herein.
BACKGROUND OF THE INVENTION
[0003] Human cells have a variety of receptors on their surfaces. G
rotein-coupled receptors ("GPCR" or "GPCRs") form one of the
largest protein families of transmembrane receptors. The human
genome has approximately 30,000 genes, as many as 1,000 of which
encode GPCRs. GPCRs have been grouped into five classes. The first
class is the rhodopsin receptor family or "Class A GPCR" with 670
receptor proteins. The rhodopsin receptor family can react with
various ligands including amines (alpha group), peptides (beta
group), lipid-like substances (gamma group), nucleotides, and
glycoproteins (delta group), and comprises a lot of drug target
receptors. The second class is the secretin receptor family, and
has binding domains for peptide hormones. Receptors in this family
are associated with homeostasis and have been arising as important
targets for drug development. The third class is the adhesion
receptor family, characterized by a GPCR proteolytic site (GPS).
Development of drugs targeting this family of GPCRs has not yet
taken place because they exhibit various N-terminal moieties and
little is known about their ligands. The fourth class is the
glutamate receptor family with 22 GPCR members have so far been
identified. Relatively little is known about the specificity of
each protein. The last class is the Frizzled/Taste2 family that
encompasses 10 Frizzled receptors for which Wnt glycoproteins serve
as ligands, 5 SMO (smoothened) receptors which need no ligands, and
25 Taste2 receptors which are required for sensing various tastes.
Receptors including GPCRs are also classified on the basis of the
identification of endogenous ligands. Receptors bind with known
endogenous compounds or are classified as orphan receptors whose
endogenous ligands have not yet been identified.
[0004] GPCRs are found in a broad range of tissue and cell types
and associated with many different physiological mechanisms. They
are activated by a wide range of ligands, e.g., hormones such as
thyroid-stimulating hormone (TSH), adrenocorticotropic hormone,
glucagon and vasopressin, amines such as 5-HT, acetylcholine
(muscarinic AchR), and histamines, lipids such as LPA and S1P, and
signal transmitters such as amino acids, Ca.sup.2+, nucleic acids,
peptides and light. The wide distribution and diversity of roles
that GPCRs play is evidence of their importance in various
pathological diseases. Indeed, GPCRs are involved in various
diseases including bronchoconstriction, hypertension, diabetes,
inflammation, cell death, hormone disorders, cancer,
neurotransmission and behavioral disorders. GPCRs are therefore an
important area for the development of pharmaceutical products.
Approximately 360 GPCRs are now considered available for drug
development. Of these, 46 have already been used for drug
development. There are approximately an estimated 150 Orphan GPCRs
(oGPCRs). In the drug development field, cell membrane receptors
act as selective sites for drug action and are responsible for 50%
of all drug targets; GPCR activity modulating drugs account for 30%
of the most frequently used top 100 drugs (40 billion dollars, 9%
of the total drug market). Therefore, GPCRs are among the most
significant targets for new drug development.
[0005] GPCRs have common structural features. They have seven
hydrophobic membrane-spanning domains, each 20-30 amino acids long,
which are connected by hydrophilic amino acid sequences of various
lengths. The receptors have an extracellular N-terminus while the
C-terminus is located in the cytoplasm. GTP-binding proteins (G
proteins) act as mediators transmitting to intracellular effectors
the signals that are generated by binding hormones or other
chemical ligands that stimulate GPCR. After ligand binding, the
GPCR intracellular domain undergoes a conformational change to
allow the receptor to interact with a G protein, which in turn
activates intracellular signal transmitters such as adenylate
cyclase, phospholipase C or ion channels. This system generates a
signaling cascade in which many secondary transmitters act in
response to the binding of one ligand to GPCR. Cells use this
mechanism to detect extracellular environmental changes and to
properly react in response to the changes. On the whole, endogenous
ligands activate receptors with the concomitant generation of a
conformational change, which allows association between the
receptors and G proteins. Recent studies on the interaction between
proteins have revealed that GPCRs associate with various proteins
such as GRK or SH2 (Src Homology 2) domain-containing proteins, and
adaptor Grb2 as well as G protein to participate in signaling
transduction.
[0006] Under normal conditions, signaling transduction brings about
the final result which is cell activation or suppression. In a
physiological environment, GPCRs exist in equilibrium between their
inactive and active states in the cell membrane. Inactive receptors
cannot exert a biological response in conjunction with cellular
signal transduction pathways. The receptors exhibit biological
responses via a signal transduction pathway (through G proteins)
only when they have structurally changed to their active form. The
receptor may be stabilized into an active form by compounds such as
endogenous ligands or drugs. Therefore, functional studies, such as
cloning such gene families, and the identification of new ligands
thereof, have the same meaning as the development of new drug
candidates, that is, siRNA, antibodies, polypeptides, effectors,
inhibitors, agonists, antagonists.
[0007] Development, differentiation, homeostasis, responses to
stimuli, control of the cell cycle, as well as the aging and
apoptosis of living organisms are mostly a result of selective
expression of specific genes within cells. This is true for
cellular mechanisms associated with diseases. Particularly,
pathological phenomena, such as oncogenesis, are induced by gene
mutations that in the end lead to changes in gene expression.
[0008] ONC201
(7-benzyl-4-(2-methylbenzyl)-1,2,6,7,8,9-hexahydroimidazo[1,2-a]pyrido
[3,4-e]pyrimidin-5(1H)-one) is the founding member of a class of
anti-cancer compounds called imipridones that is in Phase II
clinical trials in multiple advanced cancers. Since the discovery
of ONC201 as a p53-independent inducer of TRAIL gene transcription,
preclinical studies have determined that ONC201 has
anti-proliferative and pro-apoptotic effects against a broad range
of tumor cells but not normal cells. The mechanism of action of
ONC201 involves engagement of PERK-independent activation of the
integrated stress response, leading to tumor upregulation of DR5
and dual Akt/ERK inactivation, and consequent Foxo3a activation
leading to upregulation of the death ligand TRAIL. ONC201 is orally
active with infrequent dosing in animal models, causes sustained
pharmacodynamic effects, and is not genotoxic. The first-in-human
ONC201 clinical trial in advanced aggressive refractory solid
tumors confirmed that it is well-tolerated. In summary, the
imipridone family that comprises ONC201 and its chemical analogs
represent a new class of therapeutic agents.
BRIEF SUMMARY OF THE INVENTION
[0009] In one aspect, provided herein are compounds of formula
(10):
##STR00001##
wherein R.sub.1 and R.sub.2 are independently selected from H,
alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl,
heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl,
heteroarylalkyl, alkoxyalkyl, alkoxycarbonyl, aralkoxy,
aralkylthio, and acyl radicals. In one embodiment, when R.sub.1 is
CH.sub.2Ph, R.sub.2 is not CH.sub.2-(2-CH.sub.3-Ph). In one
embodiment, R.sub.1 is CH.sub.2Ph and R.sub.2 is
CH.sub.2-(2-CH.sub.3-Ph) (i.e., ONC201). In one embodiment, R.sub.1
is CH.sub.2Ph and R.sub.2 is CH.sub.2-(2,4-di F-Ph) (i.e., ONC206).
In one embodiment, R.sub.1 is CH.sub.2Ph and R.sub.2 is
CH.sub.2-(4-CF.sub.3-Ph) (i.e., ONC212). In one embodiment, R.sub.1
is CH.sub.2Ph and R.sub.2 is CH.sub.2-(3,4-di F-Ph) (i.e., ONC213).
In one embodiment, R.sub.1 is CH.sub.2 (3,4-di-Cl-Ph and R.sub.2 is
CH.sub.2-(4-CF.sub.3-Ph) (i.e., ONC234). In one embodiment, R.sub.1
is CH.sub.2-3-thienyl and R.sub.2 is CH.sub.2-(4-CF.sub.3-Ph)
(i.e., ONC236).
[0010] In another aspect, provided herein are methods of treating
or preventing a disease, disorder, or condition in a subject in
need thereof, comprising: administering to the subject in need of
such treatment a pharmaceutical composition comprising a
therapeutically effective amount a compound of formula (10) or an
analog thereof, or a pharmaceutically acceptable salt thereof. In
one embodiment, the compound is selected from the group consisting
of ONC201, ONC206, ONC212, ONC213, ONC234 and ONC236. In one
embodiment, the subject has, or is at risk of having, cancer. In
one embodiment, the cancer is selected from a central nervous
system tumor, a brain tumor, a peripheral nervous system tumor, a
pheochromocytoma, a paraganglioma, a neuroendocrine tumor, ewings
sarcoma, a pancreatic cancer, a prostate cancer, an endometrial
cancer, a hematological malignancy, a bone cancer, and a lymphatic
system tumor. In one embodiment, the cancer is selected from
meningioma, ependymoma, glioma, neuroblastoma, or diffuse intrinsic
pontine glioma. In one embodiment, the cancer is selected from an
acute leukemia selected from an acute lymphotyte leukemia, acute
myeloid leukemia, myelodysplastic syndrome, or myeloproliferative
disease. In one embodiment, In one embodiment, the cancer has a
histone H3 mutation (e.g., the mutation H3.3 K27M) or an
epigentically silenced unmethylated O(6)-methylguanine-DNA
methyltransferase (MGMT) gene. In one embodiment, the subject has,
or is at risk of having, a psychiatric disorder. In one embodiment,
the psychiatric disorder is selected from a psychosis,
schizophrenia, bipolar disorder, or major depressive disorder. In
one embodiment, the subject has, or is at risk of having, an
infection. In one embodiment, the infection is a bacterial
infection. In one embodiment, the infection is a gram-negative
bacterial infection. In one embodiment, the infection is a
gram-positive bacterial infection. In one embodiment, the bacterial
infection is an infection of a bacteria selected from the group
consisting of Enterococcus faecium, Staphylococcus aureus,
Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas
aeruginosa, and Enterobacter species. In one embodiment, the
bacterial infection is a Staphylococcus infection. In one
embodiment, the Staphylococcus infection is an S. aureus infection
(e.g., a methicillin-resistant S. aureus (MRSA) infection).
[0011] In another aspect, provided herein are methods of treating
or preventing a disease, disorder, or condition in a subject in
need of selective modulation of the activity of a G protein-coupled
receptor (GPCR) or of a G protein-coupled receptor (GPCR) signaling
pathway. Modulation includes, but is not limited to, agonism,
partial agonism, inverse agonism, partial antagonism, antagonism,
bivalent modulation, or bitopic modulation. In one embodiment, the
methods comprise administering to the subject in need of such
treatment a pharmaceutical composition comprising a therapeutically
effective amount a compound of formula (10) or an analog thereof,
or a pharmaceutically acceptable salt thereof. In one embodiment,
the subject has, or is at risk of having, cancer. In one
embodiment, the subject has, or is at risk of having, a psychiatric
disorder. In one embodiment, the psychiatric disorder is psychosis.
In one embodiment, the psychiatric disorder is schizophrenia. In
one embodiment, the subject has, or is at risk of having, an
infection. In one embodiment, the infection is a bacterial
infection. In one embodiment, the infection is a gram-negative
bacterial infection. In one embodiment, the infection is a
gram-positive bacterial infection. In one embodiment, the bacterial
infection is an infection of a bacteria selected from Enterococcus
faecium, Staphylococcus aureus, Klebsiella pneumoniae,
Acinetobacter baumannii, Pseudomonas aeruginosa, or Enterobacter
species. In one embodiment, the bacterial infection is a
Staphylococcus infection. In one embodiment, the Staphylococcus
infection is an S. aureus infection (e.g., a methicillin-resistant
S. aureus (MRSA) infection). In one embodiment, the treatment
regimen comprises administering an effective amount of a
therapeutic, such as compound of formula (10), a pharmaceutically
acceptable salt thereof, or an analog thereof. In one embodiment,
the GPCR is a Class A GPCR. In one embodiment, the GPCR is GPR132,
GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7 or a
combination thereof. In one embodiment, the GPCR is GPR132 (also
called G2A). In one embodiment, the GPCR is GPR91. In one
embodiment, the GPCR is MTNR1A. In one embodiment, the GPCR is
CXCR7.
[0012] In another aspect, provided herein are methods of treating
or preventing a disease, disorder, or condition in a subject in
need of selective modulation of the activity of a dopamine receptor
or of a member of a dopamine receptor signaling pathway. In one
embodiment, the methods comprise administering to the subject in
need of such treatment a pharmaceutical composition comprising a
therapeutically effective amount a compound of formula (10) or an
analog thereof, or a pharmaceutically acceptable salt thereof. In
one embodiment, the subject has, or is at risk of having, cancer.
In one embodiment, the subject has, or is at risk of having, a
psychiatric disorder. In one embodiment, the psychiatric disorder
is psychosis. In one embodiment, the psychiatric disorder is
schizophrenia. In one embodiment, the subject has, or is at risk of
having, an infection. In one embodiment, the infection is a
bacterial infection. In one embodiment, the infection is a
gram-negative bacterial infection. In one embodiment, the infection
is a gram-positive bacterial infection. In one embodiment, the
bacterial infection is an infection of a bacteria selected from the
group consisting of Enterococcus faecium, Staphylococcus aureus,
Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas
aeruginosa, and Enterobacter species. In one embodiment, the
bacterial infection is a Staphylococcus infection. In one
embodiment, the Staphylococcus infection is an S. aureus infection
(e.g., a methicillin-resistant S. aureus (MRSA) infection). In one
embodiment, the treatment regimen comprises administering an
effective amount of a therapeutic, such as compound of formula
(10), a pharmaceutically acceptable salt thereof, or an analog
thereof. In one embodiment, the dopamine receptor is from the
D2-like family of dopamine receptors.
[0013] In another aspect, provided herein are methods of treating
or preventing liver fibrosis or of regenerating liver tissue,
comprising: administering to the subject in need of such treatment
a pharmaceutical composition comprising a therapeutically effective
amount a compound of formula (10) or a compound of formula (100)
(e.g., TIC-10), or an analog thereof, or a pharmaceutically
acceptable salt thereof. In one embodiment, the compound is a CXCR7
agonist.
[0014] In another aspect, provided herein are methods of
stimulating the immune system (e.g., activating NK cells) in a
subject in need thereof, comprising: administering to the subject a
pharmaceutical composition comprising a therapeutically effective
amount a compound of formula (10) or an analog thereof, or a
pharmaceutically acceptable salt thereof. In one embodiment, the
compound is a GPR91 agonist. In one embodiment, the compound is
ONC213. In one embodiment, the subject has cancer and the method is
a method of cancer immunotherapy. In one embodiment, the subject
has a viral infection (e.g., HIV). In one embodiment, the subject
has systemic lupus erythematosus. In one embodiment, the method
further comprises administering a vaccine (e.g., a cancer vaccine)
to the subject, and the compound is administered as an
adjuvant.
[0015] In another aspect, provided herein are methods of
identifying whether a subject having a condition is likely to be
responsive to a treatment regimen described herein. In one
embodiment, the methods comprise (i) obtaining a biological sample
from the subject; (ii) measuring expression levels of at least one
dopamine receptor or G protein-coupled receptor (GPCR) in the
sample; (iii) comparing the levels measured in the sample to those
for a pre-determined standard; and (iv) determining whether the
subject is likely to be responsive to the treatment regimen, based
on the levels measured in the sample to those for the
pre-determined standard. In one embodiment, the subject has, or is
at risk of having, cancer. In one embodiment, the subject has, or
is at risk of having, a psychiatric disorder. In one embodiment,
the subject has, or is at risk of having, an infection. In one
embodiment, the treatment regimen further comprises administering
an effective amount of a therapeutic, such as compound of formula
(10), a pharmaceutically acceptable salt thereof, or an analog
thereof. In one embodiment, the dopamine receptor is from the
D2-like family of dopamine receptors. In one embodiment, the GPCR
is a Class A GPCR. In one embodiment, the GPCR is GPR132, GPR91,
MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7 or a combination
thereof. In one embodiment, the GPCR is GPR132, GPR91, MTNR1A,
CXCR7 or a combination thereof. In one embodiment, the GPCR is
GPR132.
[0016] In another aspect, provided herein are methods of assessing
the effectiveness of a treatment regimen described herein,
monitoring, or providing a prognosis for a subject with a
condition. In one embodiment, the methods comprises (i) obtaining a
biological sample from the subject; (ii) measuring expression
levels of at least one dopamine receptor or G protein-coupled
receptor (GPCR) in the sample; (iii) comparing the levels measured
in the sample to those for a pre-determined standard; and (iv)
determining a prognosis or determining whether the subject is
responsive to the treatment regimen, based on the levels measured
in the sample to those for the pre-determined standard. In one
embodiment, the methods comprises (i) obtaining a biological sample
from the subject; (ii) measuring gene copy number or mutations in
at least one dopamine receptor in the sample; (iii) comparing the
copy number measured or mutations found in the sample to those for
a pre-determined standard; and (iv) determining whether the subject
is responsive to the treatment regimen, based on the copy number
measured or mutations found in the sample to those for the
pre-determined standard. In one embodiment, the subject has, or is
at risk of having, cancer. In one embodiment, the subject has, or
is at risk of having, a psychiatric disorder. In one embodiment,
the subject has, or is at risk of having, an infection. In one
embodiment, the treatment regimen comprises administering an
effective amount of a therapeutic, such as compound of formula
(10), a pharmaceutically acceptable salt thereof, or an analog
thereof. In one embodiment, the dopamine receptor is selected from
DRD2, DRD2S, DRD2L, and DRD3. In one embodiment, the dopamine
receptor is from the D2-like family of dopamine receptors In one
embodiment, the GPCR is a Class A GPCR. In one embodiment, the GPCR
is GPR132, GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7
or a combination thereof.
[0017] In another aspect, provided herein are methods for screening
a potential therapeutic for a condition. In one embodiment, the
method comprises (i) contacting at least one G protein-coupled
receptor (GPCR) with a test molecule suspected of being a
therapeutic for a condition; (ii) measuring the binding affinity,
interaction or GPCR signalling of the test compound to the GPCR;
and (iii) comparing the binding affinity, interaction or signalling
of the test molecule to a pre-determined threshold. In one
embodiment, GPCR modulation or GPCR signaling modulation by the
test molecule comparable to or greater than the threshold is
indicative of a therapeutic for the condition. In one embodiment,
the condition is cancer. In one embodiment, the pre-determined
threshold is the GPCR modulation or GPCR signaling modulation of a
therapeutic, such as a compound of formula (10) or a
pharmaceutically acceptable salt thereof, or an analog thereof. In
one embodiment, the GPCR is a Class A GPCR. In one embodiment, the
GPCR is GPR132. In one embodiment, the GPCR is GPR132, GPR91,
MTNR1A, GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7 or a combination
thereof. In one embodiment, the GPCR is GPR132. In one embodiment,
the GPCR is GPR91. In one embodiment, the GPCR is MTNR1A. In one
embodiment, the GPCR is CXCR7.
[0018] In another aspect, provided herein are methods for screening
a potential therapeutic for a condition. In one embodiment, the
method comprises (i) contacting at least one dopamine receptor with
a test molecule suspected of being a therapeutic for a condition;
(ii) measuring the binding affinity, interaction or signalling of
the test molecule to the at least one dopamine receptor; and (iii)
comparing the binding affinity or interaction of the test molecule
to a pre-determined threshold. In one embodiment, modulation of the
dopamine receptor by the test molecule comparable to or greater
than the threshold is indicative of a therapeutic for the
condition. In one embodiment, the condition is cancer. In one
embodiment, the dopamine receptor is a member of the D2-like family
of dopamine receptors. In one embodiment, the pre-determined
threshold is the modulation of the dopamine receptor or dopamine
receptor signalling by a therapeutic, such as a compound of formula
(10) or a pharmaceutically acceptable salt thereof, or an analog
thereof.
[0019] In another aspect, provided herein are methods for screening
a potential therapeutic for a condition. In one embodiment, using a
processor, the method comprises (i) using a computational docking
method to model binding or interaction, if any, of one or more
3-dimensional structures (conformations) of a test molecule
suspected of being a therapeutic for the condition to a
3-dimensional structure or model of at least one dopamine receptor;
(ii) using the computational method to estimate the binding
affinity or interaction of the test molecule structure to the
structure or model of the at least one dopamine receptor; and (iii)
using the computational method to compare the binding affinity or
interaction of the test molecule to a pre-determined threshold,
wherein modulation of the dopamine receptor by the test molecule
comparable to or greater than the threshold is indicative of a
therapeutic for the condition. In one embodiment, the condition is
cancer. In one embodiment, the dopamine receptor is a member of the
D2-like family of dopamine receptors.
[0020] In another aspect, provided herein are methods of treating a
subject having a condition. In one embodiment, the method comprises
administering an effective amount of a therapeutic agent that
targets at least one dopamine receptor or G protein-coupled
receptor (GPCR). In one embodiment, the therapeutic agent is a
neutralizing agent. In one embodiment, the therapeutic agent is an
antagonist of the receptor. In one embodiment, the therapeutic
agent is an agonist of the receptor. In one embodiment, the
therapeutic agent is a competitive inhibitor of the receptor with
respect to dopamine In one embodiment, the therapeutic agent is a
non-competitive inhibitor of the receptor with respect to dopamine
In one embodiment, the therapeutic agent is selective for the
D2-like family of dopamine receptors with respect to the D1-like
family of dopamine receptors. In one embodiment, the subject has,
or is at risk of having, cancer. In one embodiment, the subject
has, or is at risk of having, a psychiatric disorder. In one
embodiment, the subject has, or is at risk of having, an infection.
In one embodiment, the dopamine receptor is a member of the D2-like
family of dopamine receptors. In one embodiment, the GPCR is a
Class A GPCR. In one embodiment, the GPCR is GPR132. In one
embodiment, the GPCR is GPR91. In one embodiment, the GPCR is
MTNR1A. In one embodiment, the GPCR is CXCR7. In one embodiment,
the GPCR is GPR132, GPR91, MTNR1A, GPR162, GPR137, BAI3, LGR4,
PTGIR, CXCR7 or a combination thereof. In one embodiment, the
therapeutic agent is a monoclonal antibody (e.g., a chimerized or
humanized monoclonal antibody), polyclonal antibody (e.g., a
chimerized or humanized polyclonal antibody), or a bispecific
antibody. In one embodiment, the therapeutic agent is a drug or
active agent, such as an anti-cancer agent, conjugated to an
antibody. In one embodiment, the therapeutic agent is a
radioactively-conjugated antibody or a small molecule-conjugated
antibody. In one embodiment, the therapeutic agent is a vector that
expresses a recombinant antibody to the dopamine receptor or GPCR.
In one embodiment, the therapeutic agent is a fusion protein or a
peptide that targets the dopamine receptor or GPCR. In one
embodiment, the therapeutic agent is an siRNA, shRNA, or an
antisense oligonucleotide that targets the dopamine receptor or
GPCR. In one embodiment, the dopamine receptor or GPCR is targeted
by CRISPR interference.
[0021] In another aspect, provided herein are methods of treating
and assessing the efficacy of a treatment in a subject having a
condition. In one embodiment, the method comprises (i) treating the
subject according to a treatment method described herein (ii)
assessing as described herein the treatment's efficacy. In one
embodiment, the subject has, or is at risk of having, cancer. In
one embodiment, the treatment regimen comprises administering an
effective amount of a therapeutic, such as a compound of formula
(10) or a pharmaceutically acceptable salt thereof or an analog
thereof. In one embodiment, the dosage of a therapeutic
administered, the frequency of administration of the compound
(e.g., a compound of formula (10)), or both, is selected or
adjusted based on the levels of gene expression or gene copy number
measured or mutations found.
BRIEF DESCRIPTION OF THE DRAWINGS
[0022] The above summary, as well as the following detailed
description of embodiments of the invention, will be better
understood when read in conjunction with the appended drawings. It
should be understood, however, that the invention is not limited to
the precise arrangements and instrumentalities shown. In the
drawings:
[0023] FIG. 1. Antagonism of dopamine receptors (DRD1, DRD2S,
DRD2L, DRD3, DRD4, and DRD5) by ONC201.
[0024] FIG. 2 illustrates soluble prolactin detected by ELISA in
the peripheral blood of advanced solid tumor patients at baseline
and following a single ONC201 dose (PO 125-625 mg). Sampling time
points post-treatment include 6 hours, 1, 2, 7, and 21 days
post-treatment.
[0025] FIG. 3. Tumor type sensitivity of the Genomic of Drug
Sensitivity in Cancer program (GDSC) cell line collection. The
average sensitivity was determined by average estimated IC.sub.50
values from cell viability assays conducted at 72 hours
post-treatment. Numbers above the bar indicates indicate the number
of cell lines per tumor type.
[0026] FIGS. 4A, 4B, 4C, and 4D. ONC201 is a selective DRD2
antagonist. (4A) Agonism of orphan or known GPCRs or antagonism of
known GPCRs using an arrestin recruitment reporter assay (10 .mu.M
ONC201). (4B) Antagonism of ligand-stimulated dopamine receptors by
ONC201 using an arrestin recruitment reporter assay. Schild
analysis of DRD2L antagonism by ONC201 using (4C) arrestin
recruitment or (4D) cAMP modulation reporters.
[0027] FIGS. 5A, 5B, 5C, 5D, 5E, and 5F. ONC201 antagonism of DRD2
is highly specific among GPCRs and other cancer drug targets. (5A)
Antagonism of GPCRs using an arrestin recruitment reporter assay
(10 .mu.M ONC201). Competition of ONC201-mediated antagonism of
DRD2L by dopamine in (5B) arrestin recruitment or (5C) cAMP
modulation reporters. (5D) Antagonism or agonism of nuclear hormone
receptors by ONC201 (2 or 20 .mu.M) with a nuclear translocation
reporter assay. (5E) In vitro inhibition of kinase enzymatic
activity by ONC201 (1 .mu.M). (5F) DRD2L antagonistic activity of
ONC201 or an ONC201 linear isomer with no biological activity using
an arrestin recruitment reporter assay.
[0028] FIGS. 6A and 6B. GBM cell lines with higher DRD2 expression
are more responsive to ONC201. (6A) Inhibition of NCI60 GBM cell
lines as a function of ONC201 concentration. (6B) Log ONC201
GI.sub.50 (M) vs DRD2 expression for each GBM cell line.
R.sup.2=0.8707.
[0029] FIG. 7. ONC201 exhibits superior selectivity among GPCRs for
DRD2 compared to other DRD2 antagonists, such as risperidone.
[0030] FIG. 8. ONC201 has a higher selectivity for tumor cells than
the antipsychotic DRD2 antagonist, thioridazine.
[0031] FIGS. 9A and 9B. Optimization of ONC201 inhibition of DRD2
calcium flux. HEK-293T cells were transfected with expression
constructs for wild-type DRD2 (9A) or a control GPCR (9B).
DRD2-specific calcium flux inhibition was investigated at 0.1 and 1
nM dopamine, for ONC201 concentrations between 100 .mu.M and 100
.mu.M. 100 .mu.M ONC201 completely inhibited DRD2 dopamine-induced
calcium flux but had no effect on the control GPCR.
[0032] FIG. 10. Comparison of DRD2 inhibitors. DRD2-specific
calcium flux inhibition was investigated at 1 nM dopamine, using
inhibitors spiperone (squares), domperidone (triangles), and ONC201
(circles) at a range of concentrations. Data for individual assays
was normalized using the no-inhibitor value (shown as 10.sup.-11 M)
as 100% activity.
[0033] FIGS. 11A and 11B. Identification of DRD2 residues critical
for dopamine-induced calcium flux. (11A) Dopamine-induced calcium
flux was assayed as before at 1 nM dopamine, across the entire DRD2
alanine-scan library. The data represent the average of three
experiments. Mutant clones were considered to be deficient for
calcium flux if they demonstrated flux values less than 2 standard
deviations below the average calcium flux value (AV-2SD) for the
entire library. (11B) The locations of the 28 mutated residues
identified are indicated (green spheres) on the DRD3 crystal
structure (PDB id 3PBL; Chien, E. Y. et al. (2010) Science
330:1091-5). The D2R/D3R antagonist eticlopride is shown in
cyan.
[0034] FIGS. 12A and 12B. Identification of DRD2 residues critical
for ONC201 inhibition of dopamine-induced calcium flux. (12A)
Dopamine-induced calcium flux was assayed as before at 1 nM
dopamine but in the presence of 100 .mu.M ONC201, across the entire
DRD2 alanine-scan library. The data represent the average of three
experiments normalized to the value for flux value with wild-type
DRD2 (% WT). Mutant clones were considered to be critical for
ONC201 inhibition if they demonstrated flux values greater than 2
standard deviations above the average calcium flux value (AV+2SD)
for the entire library. (12B) The locations of the 8 mutated
residues identified are indicated (red spheres) on the DRD3 crystal
structure.
[0035] FIG. 13. A reference compound, (+) Butaclamol, and a test
compound, ONC201 dihydrochloride, successfully competed for
[.sup.3H]Methylspiperone, with IC.sub.50 values of 2.5 nM and 21
.mu.M, respectively.
[0036] FIG. 14. Association kinetic curves for ONC201
dihydrochloride to DRD2S receptor to determine K.sub.on and
K.sub.off.
[0037] FIG. 15. Compound activity with the selected GPCR and Orphan
GPCR Biosensor Assays. Compound was tested in antagonist and
agonist mode with the desired GPCR and Orphan GPCR Biosensor
Assays. For agonist assays, data was normalized to the maximal and
minimal response observed in the presence of control ligand and
vehicle. For antagonist assays, data was normalized to the maximal
and minimal response observed in the presence of EC.sub.80 ligand
and vehicle. The following EC.sub.80 concentrations were used: CCR4
Arrestin: 0.0078 .mu.M CCL22; CHRM2 Arrestin: 26 .mu.M
Acetylcholine; and MC4R Arrestin: 0.0026 .mu.M Melanotan II.
[0038] FIG. 16. ONC206 and ONC212 demonstrated anti-cancer efficacy
across various tumor types in the NCI60 cancer cell line panel.
ONC203 is an inactive negative control
[0039] FIG. 17. ONC206 is an imipridone with improved DRD2
antagonism. ONC206, an analog of ONC201, exhibits superior
antagonism of D2-like dopamine receptor family, and retains highly
selective antagonism of D2-like dopamine receptors compared to
other antipsychotics, such as a haloperidol.
[0040] FIG. 18. Bone cancer is more responsive to ONC206 than
ONC201.
[0041] FIG. 19. Ewing's sarcoma is the most ONC206 responsive bone
cancer subtype.
[0042] FIG. 20. ONC206 anti-cancer efficacy is in the nanomolar
range in 14 out of 16 Ewing's sarcoma cell lines. ONC206
demonstrated superior efficacy compared to ONC201 in all cell
lines
[0043] FIG. 21. The imipridone ONC212 targets an orphan GPCR It is
a highly selective agonist of the orphan GPCR tumor suppressor
GPR132, and it does not engage DRD2.
[0044] FIG. 22. ONC212 induced cell death in cancer cells (HCT116)
but not normal cells (MRCS) at nanomolar concentrations.
[0045] FIG. 23. ONC212 induces the integrated stress response and
inhibits Akt/ERK phosphorylation at nanomolar concentrations and at
earlier time points compared to ONC201.
[0046] FIG. 24. ONC212 demonstrates oral and IP anti-cancer
efficacy in xenograft mouse models of colorectal and breast
cancer.
[0047] FIG. 25. Leukemia is more responsive to ONC212 than
ONC201.
[0048] FIG. 26. ONC212 demonstrates anti-cancer efficacy (and
superior efficacy compared to ONC201) in the nanomolar range in 55
leukemia cell lines regardless of subtype.
[0049] FIG. 27. GPCRs agonized or antagonized (>50%) by 9
imipridones tested. Imipridones selectively target rhodopsin-like
Class A GPCRs.
[0050] FIGS. 28A and 28B. Case study of a subject with recurrent
glioblastoma (Example 16). (28A) Tumor size relative to baseline
(%) of total tumor burden in the subject. One cycle is 3 weeks.
(28B) Contrast MRI scans at baseline, 21, 27 and 36 weeks
post-ONC201 initiation of one of 2 malignant lesions.
[0051] FIGS. 29A, 29B and 29C. ONC212 demonstrates anti-cancer
effects in acute myeloid leukemia (AML) cell lines. (29A)
Comparison of cell viability of MV411 AML cells treated with ONC212
or cytarabine. (29B) Comparison of cell viability of MOLM14, MV411
AML cells, MRCS lung fibroblasts and Hs27a bone marrow cells
treated with ONC212. (29C) Cell viability of MOLM14 and MV411 AML
cells treated with ONC212 (250 nM) for 4, 8, 24, 48, 72 and 96
h.
[0052] FIGS. 30A, 30B, and 30C. ONC212 efficacy in ONC201-resistant
AML xenograft model (MV411 AML cells (5.times.10.sup.6)
subcutaneously implanted in the flanks of athymic nude mice).
ONC212 and ONC201 were administered orally (PO) as indicated. Tumor
volume (30A and 30B) and body weight (30C) (n=10) was measured on
indicated days. * represents p<0.05 relative to vehicle.
[0053] FIG. 31. ONC206 efficacy in Ewing's sarcoma xenograft model
(MHH-ES-1 Ewing's sarcoma cells (5.times.10.sup.6) subcutaneously
implanted in the flanks of athymic nude mice). ONC206 (PO) and
methotrexate (IV) were administered on day 1 and day 13 as
indicated. Tumor volume (A) and body weight (B) (n=4) was measured
on indicated days.
[0054] FIG. 32. ONC213 (10 .mu.M) GPCR profile using a
.beta.-arrestin recruitment reporter assay.
[0055] FIG. 33. ONC213 demonstrated in vitro anti-cancer potency in
HCT116/RPMI8226 cancer cells similar to ONC212, but in vitro
toxicity to normal cells was reduced compared to ONC212.
[0056] FIG. 34. ONC237 (10 .mu.M) GPCR profile using a
.beta.-arrestin recruitment reporter assay.
[0057] FIG. 35. ONC236 (10 .mu.M) GPCR profile using a
.beta.-arrestin recruitment reporter assay.
[0058] FIG. 36. ONC234 (10 .mu.M) GPCR profile using a
.beta.-arrestin recruitment reporter assay.
[0059] FIG. 37. ONC201 linear isomer (TIC-10) (10 .mu.M) GPCR
profile using a .beta.-arrestin recruitment reporter assay.
[0060] FIG. 38. Number of GPCRs hit for several imipridones.
DETAILED DESCRIPTION OF THE INVENTION
[0061] Scientific and technical terms used here are intended to
have the meanings commonly understood by those of ordinary skill in
the art. Such terms are found and used in context in various
standard references illustratively including J. Sambrook and D. W.
Russell, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor
Laboratory Press; 3.sup.rd Ed., 2001; F. M. 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; D. L. Nelson and M. M. Cox, Lehninger Principles of
Biochemistry, 4th Ed., W.H. Freeman & Company, 2004; Engelke,
D. R., RNA Interference (RNAi): Nuts and Bolts of RNAi Technology,
DNA Press LLC, Eagleville, P A, 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, 3.sup.rd 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, as well as U.S. Pat. No. 8,673,923. The
content of each of the references above is hereby incorporated by
reference in its entirety.
[0062] The term "substituted" means that any one or more hydrogens
on the designated atom is replaced with a selection from the
indicated group, provided that the designated atom's normal valency
is not exceeded, and that the substitution results in a stable
compound. When a substituent is keto (i.e., .dbd.O), then 2
hydrogens on the atom are replaced. Keto substituents are not
present on aromatic moieties. Ring double bonds are double bonds
that are formed between two adjacent ring atoms (e.g., C.dbd.C,
C.dbd.N, or N.dbd.N).
[0063] When a variable (e.g., R.sup.4) occurs more than one time in
a constituent or formula for a compound, its definition at each
occurrence is independent of its definition at every other
occurrence. Thus, for example, if a group is shown to be
substituted with 0-3 R.sup.4 moieties, then the group may
optionally be substituted with up to three R.sup.4 moieties and
R.sup.4 at each occurrence is selected independently from the
definition of R.sup.4. Also, combinations of substituents and/or
variables are permissible, but only if such combinations result in
stable compounds.
[0064] When an atom or chemical moiety is followed by a subscripted
numeric range (e.g., C.sub.1-6), it will be appreciated that this
is meant to encompass each number within the range, as well as all
intermediate ranges. For example, "C.sub.1-6 alkyl" is meant to
include alkyl groups with 1, 2, 3, 4, 5, 6, 1-6, 1-5, 1-4, 1-3,
1-2, 2-6, 2-5, 2-4, 2-3, 3-6, 3-5, 3-4, 4-6, 4-5, and 5-6
carbons.
[0065] The term "alkyl" includes both branched and straight-chain
saturated aliphatic hydrocarbon groups having the specified number
of carbon atoms. For example, C.sub.1-6 alkyl is intended to
include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5, and C.sub.6
alkyl groups. Examples of alkyl include, but are not limited to,
methyl, ethyl, n-propyl, i-propyl, n-butyl, isobutyl s-butyl,
t-butyl, n-pentyl, s-pentyl, neopentyl and n-hexyl. In certain
cases, a straight chain or branched chain alkyl has six or fewer
carbon atoms in its backbone (e.g., C.sub.1-C.sub.6 for straight
chain, C.sub.3-C.sub.6 for branched chain), and in other cases, a
straight chain or branched chain alkyl has four or fewer carbon
atoms. Likewise, cycloalkyls have from three to eight carbon atoms
in their ring structure, and in other cases, cycloalkyls have five
or six carbons in the ring structure. Most preferred is C.sub.1-6
alkyl, particularly ethyl, methyl, isopropyl, isobutyl, n-pentyl,
n-hexyl and cyclopropylmethyl.
[0066] the term "substituted alkyl" means alkyl as defined above,
substituted by one, two or three substituents selected from
halogen, --OH, alkoxy, --NH.sub.2, --N(CH.sub.3).sub.2,
--C(.dbd.O)OH, trifluoromethyl, --C(.dbd.O)O(C.sub.1-C.sub.4)alkyl,
--C(.dbd.O)NH.sub.2, --SO.sub.2NH.sub.2, --C(.dbd.NH)NH.sub.2, and
--NO.sub.2, preferably containing one or two substituents selected
from halogen, --OH, alkoxy, --NH.sub.2, trifluoromethyl,
--N(CH.sub.3).sub.2, and --C(.dbd.O)OH, more preferably selected
from halogen, alkoxy and --OH. Examples of substituted alkyls
include, but are not limited to, 2,2-difluoropropyl,
2-carboxycyclopentyl and 3-chloropropyl.
[0067] Unless the number of carbons is otherwise specified, "lower
alkyl" is an alkyl group, as defined above, but having from one to
six carbon atoms, preferably one to four, in its backbone
structure. "Lower alkenyl" and "lower alkynyl" have chain lengths
of 2-6 carbon atoms and preferably 2-4 carbon atoms.
[0068] "Alkenyl" includes unsaturated aliphatic groups analogous in
length and possible substitution to the alkyls described above, but
that contain at least one double bond. For example, the term
"alkenyl" includes straight-chain alkenyl groups (e.g., ethenyl,
propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl,
decenyl), branched-chain alkenyl groups, cycloalkenyl (e.g.,
alicyclic) groups (e.g., cyclopropenyl, cyclopentenyl,
cyclohexenyl, cycloheptenyl, cyclooctenyl), alkyl or alkenyl
substituted cycloalkenyl groups, and cycloalkyl or cycloalkenyl
substituted alkenyl groups. In certain cases, a straight chain or
branched chain alkenyl group has six or fewer carbon atoms in its
backbone (e.g., C.sub.2-C.sub.6 for straight chain, C.sub.3-C.sub.6
for branched chain). Likewise, cycloalkenyl groups may have from
three to eight carbon atoms in their ring structure, and in some
embodiments, cycloalkenyl groups have five or six carbons in the
ring structure. The term "C.sub.2-C.sub.6" includes alkenyl groups
containing two to six carbon atoms. The term "C.sub.3-C.sub.6"
includes alkenyl groups containing three to six carbon atoms.
[0069] "Alkynyl" includes unsaturated aliphatic groups analogous in
length and possible substitution to the alkyls described above, but
which contain at least one triple bond. For example, "alkynyl"
includes straight-chain alkynyl groups (e.g., ethynyl, propynyl,
butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl),
branched-chain alkynyl groups, and cycloalkyl or cycloalkenyl
substituted alkynyl groups. In certain embodiments, a straight
chain or branched chain alkynyl group has six or fewer carbon atoms
in its backbone (e.g., C.sub.2-C.sub.6 for straight chain,
C.sub.3-C.sub.6 for branched chain). The term "C.sub.2-C.sub.6"
includes alkynyl groups containing two to six carbon atoms. The
term "C.sub.3-C.sub.6" includes alkynyl groups containing three to
six carbon atoms.
[0070] The term "cycloalkyl" refers to a mono cyclic or polycyclic
non-aromatic radical, where each of the atoms forming the ring
(i.e. skeletal atoms) is a carbon atom. In some cases, the
cycloalkyl group is saturated or partially unsaturated. In other
cases, the cycloalkyl group is fused with an aromatic ring.
Cycloalkyl groups include groups with from 3 to 10 ring atoms.
Examples of cycloalkyl groups include, but are not limited to, the
following moieties:
##STR00002##
[0071] Monocyclic cycloalkyls include cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Dicyclic
cycloalkyls include, but are not limited to, tetrahydronaphthyl,
indanyl, and tetrahydropentalene. Polycyclic cycloalkyls include
adamantine and norbornane. The term cycloalkyl includes
"unsaturated nonaromatic carbocyclyl" or "nonaromatic unsaturated
carbocyclyl" groups, both of which refer to a nonaromatic
carbocycle as defined herein, which contains at least one carbon
carbon double bond or one carbon carbon triple bond.
[0072] The term "cycloalkylalkyl" refers to an alkyl group
substituted by a cycloalkyl group. Example cycloalkylalkyl groups
include cyclopropylalkyl, cyclohexylalkyl.
[0073] The term "heterocycloalkyl" refers to a non-aromatic
heterocycle where one or more of the ring-forming atoms is a
heteroatom such as an O, N, or S atom. Heterocycloalkyl groups
include mono- or polycyclic (e.g., having 2, 3 or 4 fused rings)
ring systems, as well as spirocycles. Example heterocycloalkyl
groups include morpholino, thiomorpholino, piperazinyl,
tetrahydrofuranyl, tetrahydrothienyl, 2,3-dihydrobenzofuryl,
1,3-benzodioxole, benzo-1,4-dioxane, piperidinyl, pyrrolidinyl,
isoxazolidinyl, isothiazolidinyl, pyrazolidinyl, oxazolidinyl,
thiazolidinyl, and imidazolidinyl. Also included in the definition
of heterocycloalkyl can be moieties that have one or more aromatic
rings fused (i.e., having a bond in common with) to the nonaromatic
heterocyclic ring, for example quinolyl, isoquinolyl, and benzo
derivatives of heterocycles. A heterocycloalkyl group having one or
more fused aromatic rings are attached though either the aromatic
or non-aromatic portion. Also included in the definition of
heterocycloalkyl are moieties where one or more ring-forming atoms
can be substituted by 1 or 2 oxo or sulfido groups. In some cases,
the heterocycloalkyl group has from 1 to about 20 carbon atoms, and
in further case from about 3 to about 20 carbon atoms. In some
cases, a heterocycloalkyl group contains 3 to about 20, 3 to about
14, 3 to about 7, or 5 to 6 ring-forming atoms. In some cases, a
heterocycloalkyl group has 1 to about 4, 1 to about 3, or 1 to 2
heteroatoms. In some cases, a heterocycloalkyl group contains 0 to
3 double bonds. In some cases, a heterocycloalkyl group contains 0
to 2 triple bonds.
[0074] The term "heterocycloalkylalkyl" refers to an alkyl group
substituted by a heterocycloalkyl. Example heterocycloalkylalkyls
include morpholinoalkyl and piperazinylalkyl.
[0075] The term "aryl" refers to monocyclic or polycyclic (e.g.,
having 2, 3 or 4 fused rings) aromatic hydrocarbons such as,
phenyl, naphthyl, anthracenyl, phenanthrenyl. In some cases, an
aryl group has from 6 to about 20 carbon atoms.
[0076] The term "arylalkyl" refers to an alkyl group substituted by
an aryl group. Example arylalkyl groups include benzyl and
phenylethyl.
[0077] The term "heteroaryl" refers to an aromatic heterocycle
having at least one heteroatom ring member such as an O, S, or N
atom. Heteroaryl groups include monocyclic and polycyclic (e.g.,
having 2, 3 or 4 fused rings) systems. A ring-forming N atom in a
heteroaryl group can also be oxidized to form an N-oxo moiety.
Examples of heteroaryl groups include pyridyl, N-oxopyridyl,
pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furyl, quinolyl,
isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl, pyrryl,
oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl,
pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl,
isothiazolyl, benzothienyl, purinyl, carbazolyl, benzimidazolyl,
indolinyl. In some cases, a heteroaryl group has from 1 to about 20
carbon atoms, and in further cases from about 3 to 20 carbon atoms.
In some cases, a heteroaryl group contains 3 to about 14, 3 to
about 7, or 5-6 ring-forming atoms. In some cases, a heteroaryl
group has 1 to about 4, 1 to about 3, or 1-2 heteroatoms.
[0078] a "heteroarylalkyl" group refers to an alkyl group
substituted by a heteroaryl group. An example of a heteroarylalkyl
group is pyridylmethyl.
[0079] The terms "halo" or "halogen" refer to a fluorine (F),
chlorine (Cl), bromine (Br), or iodine (I) atom; preferably, F, Cl,
or Br; more preferably, F or Cl. The term "perhalogenated" refers
to a moiety where all hydrogens are replaced by halogens. The term
"haloalkyl" refers to alkyl moieties with a halogen replacing a
hydrogen on one or more carbons of the hydrocarbon backbone.
C.sub.1-C.sub.6 haloalkyl includes a straight chain or branched
alkyl with six or fewer backbone carbon atoms and a halogen
replacing a hydrogen on one or more backbone carbons.
[0080] The term "alkoxy" or "alkoxyl" includes substituted and
unsubstituted alkyl, alkenyl, and alkynyl groups covalently linked
to an oxygen atom. C.sub.1-C.sub.6 alkoxy refers to moieties having
six or fewer carbon atoms in the hydrocarbon backbone. Examples of
alkoxy groups (or alkoxyl radicals) include methoxy, ethoxy,
isopropyloxy, propoxy, butoxy, and pentoxy groups. Preferred are
(C.sub.1-C.sub.3) alkoxy, particularly ethoxy and methoxy. Examples
of substituted alkoxy groups include halogenated alkoxy groups.
[0081] The term "hydroxy" or "hydroxyl" includes groups with an
--OH or --O.sup.-.
[0082] The term "pharmaceutically acceptable salts" refers to
derivatives of compounds where the parent compound is modified by
converting an existing acid or base moiety to its salt form.
Examples of pharmaceutically acceptable salts include, but are not
limited to, mineral or organic acid salts of basic residues such as
amines; alkali or organic salts of acidic residues such as
carboxylic acids. Pharmaceutically acceptable salts include
conventional non-toxic salts of a parent compound formed, for
example, from non-toxic inorganic or organic acids.
Pharmaceutically acceptable salts may be synthesized from a parent
compound which contains a basic or acidic moiety by conventional
chemical methods. Generally, such salts can be prepared by reacting
a free acid or base form of these compounds with a stoichiometric
amount of the appropriate base or acid in water or in an organic
solvent, or in a mixture of the two; generally, nonaqueous media
like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile
are preferred. Lists of suitable salts can be found in Remington's
Pharmaceutical Sciences, 17.sup.th ed., Mack Publishing Company,
Easton, Pa., 1985, p. 1418, Journal of Pharmaceutical Science, 66,
2 (1977), and P. H. Stahl and C. G. Wermuth, editors, Handbook of
Pharmaceutical Salts: Properties, Selection and Use, 2.sup.nd
Revised edition, Weinheim/Thrich:Wiley-VCH/VHCA (2011), each of
which is incorporated herein by reference in its entirety.
[0083] Examples of suitable inorganic acids include hydrochloric
acid, sulphuric acid, phosphoric acid, or hydrobromic acid, while
examples of suitable organic acids include carboxylic acid, sulpho
acid, or sulphonic 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, benzoic
acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, cinnamic acid,
mandelic acid, citric acid, maleic acid, salicylic acid,
trifluoroacetic acid, 3-aminosalicylic acid, ascorbic acid, embonic
acid, nicotinic acid, isonicotinic acid, oxalic acid, gluconic
acid, amino acids, methanesulphonic acid, ethanesulphonic acid,
2-hydroxyethanesulphonic acid, ethane-1,2-disulphonic acid,
benzenesulphonic acid, 4-methylbenzenesulphonic acid or
naphthalene-2-sulphonic acid. Examples of suitable inorganic bases
include sodium hydroxide, potassium hydroxide and ammonia, while
examples of suitable organic bases include amines, e.g., tertiary
amines, such as trimethylamine, triethylamine, pyridine,
NN-dimethylaniline, quinoline, isoquinoline, .alpha.-picoline,
.beta.-picoline, .gamma.-picoline, quinaldine, or pyrimidine.
[0084] the term "antibody" encompasses the structure that
constitutes the natural biological form of an antibody. In most
mammals, including humans, and mice, this form is a tetramer and
consists of two identical pairs of two immunoglobulin chains, each
pair having one light and one heavy chain, each light chain
comprising immunoglobulin domains V.sub.L and C.sub.L, and each
heavy chain comprising immunoglobulin domains V.sub.H, C.gamma.1,
C.gamma.2, and C.gamma.3. In each pair, the light and heavy chain
variable regions (V.sub.L and V.sub.H) are together responsible for
binding to an antigen, and the constant regions (C.sub.L,
C.gamma.1, C.gamma.2, and C.gamma.3, particularly C.gamma.2, and
C.gamma.3) are responsible for antibody effector functions. In some
mammals, for example in camels and llamas, full-length antibodies
may consist of only two heavy chains, each heavy chain comprising
immunoglobulin domains V.sub.H, C.gamma.2, and C.gamma.3. By
"immunoglobulin (Ig)" herein is meant a protein consisting of one
or more polypeptides substantially encoded by immunoglobulin genes.
Immunoglobulins include but are not limited to antibodies
Immunoglobulins may have a number of structural forms, including
full-length antibodies, antibody fragments, and individual
immunoglobulin domains including V.sub.H, C.gamma.1, C.gamma.2,
C.gamma.3, V.sub.L, and C.sub.L.
[0085] Based on the heavy-chain constant domain amino acid
sequence, intact antibodies can be assigned to different "classes."
There are five-major classes (isotypes) of intact antibodies: IgA,
IgD, IgE, IgG, and IgM, and several of these may be further divided
into "subclasses," e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The
heavy-chain constant domains that correspond to the different
antibody classes are called alpha, delta, epsilon, gamma, and mu,
respectively. The subunit structures and three-dimensional
configurations of different classes of immunoglobulins are well
known to one skilled in the art.
[0086] The terms "antibody" or "antigen-binding fragment,"
respectively, refer to intact molecules as well as functional
fragments thereof, such as Fab, a scFv-Fc bivalent molecule,
F(ab').sub.2, and Fv that are capable of specifically interacting
with a desired target. In some cases, the antigen-binding fragments
comprise: [0087] (1) Fab, the fragment which contains a monovalent
antigen-binding fragment of an antibody molecule, which can be
produced by digestion of whole antibody with the enzyme papain to
yield an intact light chain and a portion of one heavy chain;
[0088] (2) Fab', the fragment of an antibody molecule that can be
obtained by treating whole antibody with pepsin, followed by
reduction, to yield an intact light chain and a portion of the
heavy chain; two Fab' fragments are obtained per antibody molecule;
[0089] (3) (Fab').sub.2, the fragment of the antibody that can be
obtained by treating whole antibody with the enzyme pepsin without
subsequent reduction; F(ab').sub.2 is a dimer of two Fab' fragments
held together by two disulfide bonds; [0090] (4) Fv, a genetically
engineered fragment containing the variable region of the light
chain and the variable region of the heavy chain expressed as two
chains; [0091] (5) Single chain antibody ("SCA"), a genetically
engineered molecule containing the variable region of the light
chain and the variable region of the heavy chain, linked by a
suitable polypeptide linker as a genetically fused single chain
molecule; and [0092] (6) scFv-Fc, is produced by fusing
single-chain Fv (scFv) with a hinge region from an immunoglobulin
(Ig) such as an IgG, and Fc regions.
[0093] In one embodiment, an antibody provided herein is a
monoclonal antibody. In one embodiment, the antigen-binding
fragment provided herein is a single chain Fv (scFv), a diabody, a
tandem scFv, a scFv-Fc bivalent molecule, an Fab, Fab', Fv,
F(ab').sub.2 or an antigen binding scaffold (e.g., affibody,
monobody, anticalin, DARPin, Knottin).
[0094] the terms "binds," "binding" or grammatical equivalents,
refer to compositions, directly or indirectly, having affinity for
each other. "Specific binding" is where the binding is selective
between two molecules. A particular example of specific binding
occurs between an antibody and an antigen. Typically, specific
binding can be distinguished from non-specific when the
dissociation constant (K.sub.D) is less than about
1.times.10.sup.-5M or less than about 1.times.10.sup.-6 M or
1.times.10.sup.-7 M. Specific binding can be detected, for example,
by ELISA, immunoprecipitation, coprecipitation, with or without
chemical crosslinking, and two-hybrid assays. Appropriate controls
can be used to distinguish between "specific" and "non-specific"
binding. "Affinity" is the strength of the binding interaction of
two molecules, such as an antigen and its antibody, which is
defined for antibodies and other molecules with more than one
binding site as the strength of binding of the ligand at one
specified binding site. Although the noncovalent attachment of a
ligand to antibody or other molecule is typically not as strong as
a covalent attachment, "high affinity" is for a ligand that binds
to an antibody or other molecule having an affinity constant
(K.sub.a) of greater than 10.sup.4 M.sup.-1, typically
10.sup.5-10.sup.11 M.sup.-1; as determined by inhibition ELISA or
an equivalent affinity determined by comparable techniques, such as
Scatchard plots or using K.sub.d/dissociation constant, which is
the reciprocal of the K.sub.a.
[0095] The term "selective" with respect to binding, inhibition,
stimulation, or modulation means preferential binding, inhibition,
stimulation, or modulation, respectively, of a first activity
relative to a second activity (e.g., preferential binding of one
receptor to another receptor; preferential inhibition relative to
other receptors; or preferential inhibition of a mutant to a
wild-type or vice versa). In some cases, binding is greater than
two times more selective, greater than five times more selective,
greater than ten times more selective, greater than fifty times
more selective, greater than 100 times more selective, or greater
than 1000 times more selective for the desired molecular target or
pathway versus an undesired molecular target or pathway. In some
cases, a compound will bind a first molecular target or affect a
pathway by at least 2-fold, at least 5-fold, at least 10-fold, at
least 20-fold, at least 50-fold, at least 100-fold relative to a
second target or pathway under the same conditions. It will be
appreciated that in preferred embodiments, binding to the D2-like
family of dopamine receptors or a member thereof, will be selective
with respect to the D1-like family of dopamine receptors or a
member thereof by any of the foregoing amounts. The in vitro or in
vivo activity of a molecular target or pathway may be measured by
any suitable reproducible means.
[0096] The term "modulating" refers to "stimulating" or
"inhibiting" an activity of a molecular target or pathway. For
example, a composition modulates the activity of a molecular target
or pathway if it stimulates or inhibits the activity of that target
or pathway by at least 10%, by at least about 20%, by at least
about 25%, by at least about 30%, by at least about 40%, by at
least about 50%, by at least about 60%, by at least about 70%, by
at least about 75%, by at least about 80%, by at least about 90%,
by at least about 95%, by at least about 98%, or by about 99% or
more relative to the activity of that molecular target or pathway
under the same conditions but lacking only the presence of the
composition. In another example, a composition modulates the
activity of a molecular target or pathway if it stimulates or
inhibits the activity of that target or pathway by at least 2-fold,
at least 5-fold, at least 10-fold, at least 20-fold, at least
50-fold, at least 100-fold relative to the activity of that target
or pathway under the same conditions but lacking only the presence
of the composition. The activity of a molecular target or pathway
may be measured by any reproducible means. For example, the
activity of a molecular target or pathway may be measured in vitro
or in vivo by a suitable assay known in the art for measuring the
activity. Control samples (untreated with the composition) can be
assigned a relative activity value of 100%.
[0097] In one embodiment, an antibody, antigen-binding fragment, or
affinity tag binds its target with a K.sub.D of 0.1 nM-10 mM, 0.1
nM-1 mM, or within the 0.1 nM range. In one embodiment, an
antibody, antigen-binding fragment, or affinity tag binds its
target with a K.sub.D of 0.1-2 nM, 0.1-1 nM, 0.05-1 nM, 0.1-0.5 nM,
or 0.1-0.2 nM. In one embodiment, an antibody, antigen-binding
fragment, or affinity tag bind its target directly. In one
embodiment, an antibody, antigen-binding fragment, or affinity tag
bind its target indirectly, for example, binding as a secondary
antibody that binds to an antibody bound to the target.
[0098] The word "label" refers to a compound or composition which
is conjugated or fused directly or indirectly to a reagent such as
a nucleic acid probe or an antibody and facilitates detection of
the reagent to which it is conjugated or fused. The label may
itself be detectable (e.g., radioisotope or fluorescent labels) or,
in the case of an enzymatic label, may catalyze chemical alteration
of a substrate compound or composition, which is detectable.
[0099] The term "probe" refers to synthetic or biologically
produced nucleic acids that contain specific nucleotide sequences
which hybridize under stringent conditions to target nucleic acid
sequences. The terms "labeled probe," "nucleic acid probe operably
linked to a detectable label," or "nucleic acid strand operably
linked to a detectable label" refer to a probe which is prepared
with a marker moiety or "detectable label" for detection. The
marker moiety is attached at either the 5' end, the 3' end,
internally, or a combination thereof. That is, one probe may be
attached to multiple marker moieties. A preferred moiety is an
identifying label such as a fluorophore. A labeled probe may also
comprise a plurality of different nucleic acid sequences each
labeled with one or more marker moieties. Each marker moiety may be
the same or different. It may be beneficial to label different
probes (e.g., nucleic acid sequences) each with a different marker
moiety. This can be achieved by having a single distinguishable
moiety on each probe. For example, probe A is attached to moiety X
and probe B is attached to moiety Y. Alternatively, probe A is
attached to moieties X and Y while probe B is attached to moiety Z
and W. Alternatively, probe A is attached to moieties X and Y,
while probe B is attached to moieties Y and Z. All probes "A" and
"B" above would be distinguishable and uniquely labeled.
[0100] By "tissue sample" is meant a collection of similar cells
obtained from a tissue of a subject or patient, preferably
containing nucleated cells with chromosomal material. The four main
human tissues are (1) epithelium; (2) connective tissues, including
blood vessels, bone and cartilage; (3) muscle tissue; and (4) nerve
tissue. The tissue sample source may be solid tissue as from a
fresh, frozen and/or preserved organ or tissue sample or biopsy or
aspirate; blood or a blood constituent; bodily fluids such as
cerebral spinal fluid, amniotic fluid, peritoneal fluid, or
interstitial fluid; cells from a time in gestation or development
of the subject. A tissue sample may be primary or cultured cells or
cell lines. A tissue sample may contain compounds that are not
naturally intermixed with the tissue in nature such as
preservatives, anticoagulants, buffers, fixatives, nutrients, or
antibiotics. By a "section" of a tissue sample is meant a single
part or piece of a tissue sample, e.g., a thin slice of tissue or
cells cut from a tissue sample. Multiple sections of tissue samples
may be taken and subjected to analysis. A "cell line" refers to a
permanently established cell culture that will proliferate given
appropriate fresh medium and space.
Detection Methods
[0101] In various aspects, provided herein are methods of detecting
or measuring a target receptor (e.g., a dopamine receptor or a
GPCR) in a biological sample. Targets are detected by contacting
the sample with a target detection reagent, e.g., an antibody or
fragment thereof, and a labeling reagent. The presence or absence
of targets are detected by the presence or absence of the labeling
reagent. In some instances, a sample is contacted with the target
detection and the labeling reagents concurrently e.g., the
detection reagent is a primary antibody and the labeling reagent is
a fluorescent dye conjugated to it. Alternatively, the biological
sample is contacted with the target detection and labeling reagents
sequentially, e.g., the detection reagent is a primary antibody and
the labeling reagent includes a secondary antibody. For example, a
sample is incubated with a detection reagent, in some cases
together with a labeling reagent, under conditions that allow a
complex between the detection reagent (and labeling reagent) and
target to form. After complex formation the sample is optionally
washed one or more times to remove unbound detection reagent (and
labeling reagent). When the sample is further contacted with a
labeling reagent that specifically binds the detection reagent
bound to the target, the sample can optionally be washed one or
more times to remove unbound labeling reagent. The presence or
absence of the target in the sample is then determined by detecting
the labeling reagent.
[0102] The methods described here provide for detection of multiple
targets in a sample. Multiple targets are identified by contacting
the biological sample with additional detection reagents followed
by additional labeling reagent specific for the additional
detection reagents using the methods described.
[0103] A detection moiety, i.e., detectable label, is a substance
used to facilitate identification and/or quantitation of a target.
Detection moieties are directly observed or measured or indirectly
observed or measured. Detection moieties include, but are not
limited to, radiolabels that can be measured with
radiation-counting devices; pigments, dyes or other chromogens that
can be visually observed or measured with a spectrophotometer; spin
labels that can be measured with a spin label analyzer; and
fluorescent moieties, where the output signal is generated by the
excitation of a suitable molecular adduct and that can be
visualized by excitation with light that is absorbed by the dye or
can be measured with standard fluorometers or imaging systems. The
detection moiety can be a luminescent substance such as a phosphor
or fluorogen; a bioluminescent substance; a chemiluminescent
substance, where the output signal is generated by chemical
modification of the signal compound; a metal-containing substance;
or an enzyme, where an enzyme-dependent secondary generation of
signal occurs, such as the formation of a colored product from a
colorless substrate. The detection moiety may also take the form of
a chemical or biochemical, or an inert particle, including
colloidal gold, microspheres, quantum dots, or inorganic crystals
such as nanocrystals or phosphors. The term detection moiety or
detectable label can also refer to a "tag" or hapten that can bind
selectively to a labeled molecule such that the labeled molecule,
when added subsequently, is used to generate a detectable signal.
For instance, one can use biotin, iminobiotin or desthiobiotin as a
tag and then use an avidin or streptavidin conjugate of horseradish
peroxidase (HRP) to bind to the tag, and then use a chromogenic
substrate (e.g., tetramethylbenzidine) or a fluorogenic substrate
such as Amplex Red or Amplex Gold (Molecular Probes, Inc.) to
detect the presence of HRP. Similarly, the tag can be a hapten or
antigen (e.g., digoxigenin), and an enzymatically, fluorescently,
or radioactively labeled antibody can be used to bind to the tag.
Numerous labels are known by those of skill in the art and include,
but are not limited to, particles, fluorescent dyes, haptens,
enzymes and their chromogenic, fluorogenic, and chemiluminescent
substrates.
[0104] A fluorophore is a chemical moiety that exhibits an
absorption maximum beyond 280 nm, and when covalently attached in a
labeling reagent retains its spectral properties. Fluorophores
include a pyrene, an anthracene, a naphthalene, an acridine, a
stilbene, an indole or benzindole, an oxazole or benzoxazole, a
thiazole or benzothiazole, a porphyrin, a cyanine, a perylene, a
4-amino-7-nitrobenz-2-oxa-1,3-diazole (NBD), a carbocyanine, a
carbostyryl, a salicylate, an anthranilate, an azulene, a pyridine,
a quinoline, a borapolyazaindacene, a xanthene, an oxazine or a
benzoxazine, a carbazine, a phenalenone, a coumarin, a benzofuran
and benzphenalenone and derivatives thereof. oxazines include
resorufins, aminooxazinones, diaminooxazines, and their
benzo-substituted analogs.
[0105] When the fluorophore is a xanthene, the fluorophore may be a
fluorescein, a rhodol, or a rhodamine Fluorescein includes benzo-
or dibenzofluoresceins, seminaphthofluoresceins, or
naphthofluoresceins. Similarly, rhodol includes
seminaphthorhodafluors. Alternatively, the fluorophore is a
xanthene that is bound via a single covalent bond at the 9-position
of the xanthene. Preferred xanthenes include derivatives of
3H-xanthen-6-ol-3-one, derivatives of 6-amino-3H-xanthen-3-one, or
derivatives of 6-amino-3H-xanthen-3-imine Fluorophores include
xanthene (rhodol, rhodamine, fluorescein and derivatives thereof)
coumarin, cyanine, pyrene, oxazine and borapolyazaindacene. In
addition, the fluorophore can be sulfonated xanthenes, fluorinated
xanthenes, sulfonated coumarins, fluorinated coumarins and
sulfonated cyanines. The choice of fluorophore in the labeling
reagent will determine the absorption and fluorescence emission
properties of the labeling reagent. Physical properties of a
fluorophore label include spectral characteristics (absorption,
emission and stokes shift), fluorescence intensity, lifetime,
polarization and photo-bleaching rate can all be used to
distinguish one fluorophore from another.
[0106] Typically, a fluorophore contains one or more aromatic or
heteroaromatic rings that are optionally substituted by one or more
of a variety of substituents, including halogen, nitro, cyano,
alkyl, perfluoroalkyl, alkoxy, alkenyl, alkynyl, cycloalkyl,
arylalkyl, acyl, aryl or heteroaryl ring system, benzo, or other
substituents typically present on fluorophores known in the
art.
[0107] Preferably, the detection moiety is a fluorescent dye.
Fluorescent dyes include, for example, Fluorescein, Rhodamine,
Texas Red, Cy2, Cy3, Cy5, Cy0, Cy0.5, Cy1, Cy1.5, Cy3.5, Cy7,
VECTOR Red, ELF.TM. (Enzyme-Labeled Fluorescence), FluorX, Calcein,
Calcein-AM, CRYPTOFLUOR.TM.'S, Orange (42 kDa), Tangerine (35 kDa),
Gold (31 kDa), Red (42 kDa), Crimson (40 kDa), BHMP, BHDMAP,
Br-Oregon, Lucifer Yellow, Alexa dye family,
N-(6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)caproyl) (NBD),
BODIPY.TM., boron dipyrromethene difluoride, Oregon Green,
MITOTRACKER.TM. Red, DiOC7 (3), DiIC18, Phycoerythrin,
Phycobiliproteins BPE (240 kDa) RPE (240 kDa) CPC (264 kDa) APC
(104 kDa), Spectrum Blue, Spectrum Aqua, Spectrum Green, Spectrum
Gold, Spectrum Orange, Spectrum Red, NADH, NADPH, FAD, Infra-Red
(IR) Dyes, Cyclic GDP-Ribose (cGDPR), Calcofluor White, Tyrosine
and Tryptophan. Many fluorophores can also function as chromophores
and thus they are also preferred chromophores.
[0108] In addition to fluorophores, enzymes also find use as
detectable moieties. Enzymes are desirable detectable moieties
because amplification of a detectable signal can be achieved
resulting in increased assay sensitivity. The enzyme itself does
not produce a detectable response but breaks down a substrate when
it is contacted by an appropriate substrate such that the converted
substrate produces a fluorescent, colorimetric or luminescent
signal. Enzymes amplify a detectable signal because one enzyme on a
labeling reagent can result in multiple substrates being converted
to a detectable signal. This is advantageous where there is a low
quantity of target present in the sample or a fluorophore does not
exist that will give comparable or stronger signal than the enzyme.
However, fluorophores are preferred because they do not require
additional assay steps, and thus reduce the overall time to
complete an assay. The enzyme substrate is selected to yield the
preferred measurable product, e.g. colorimetric, fluorescent or
chemiluminescence. Such substrates are extensively used in the
art.
[0109] A preferred colorimetric or fluorogenic substrate and enzyme
combination uses oxidoreductases such as horseradish peroxidase and
a substrate such as 3,3'-diaminobenzidine (DAB) and
3-amino-9-ethylcarbazol-e (AEC), which yield a distinguishing color
(brown and red, respectively). Other colorimetric oxidoreductase
substrates that yield detectable products include, but are not
limited to: 2,2-azino-bis(3-ethylbenzothiaz-oline-6-sulfonic acid)
(ABTS), o-phenylenediamine (OPD), 3,3',5,5'-tetramethylbenzidine
(TMB), o-dianisidine, 5-aminosalicylic acid, 4-chloro-1-naphthol.
Fluorogenic substrates include, but are not limited to,
homovanillic acid or 4-hydroxy-3-methoxyphenylacetic acid, reduced
phenoxazines and reduced benzothiazines, including Amplexe Red
reagent and its variants and reduced dihydroxanthenes, including
dihydrofluoresceins and dihydrorhodamines including
dihydrorhodamine 123. Peroxidase substrates that are tyramides
represent a unique class of peroxidase substrates in that they can
be intrinsically detectable before action of the enzyme but are
"fixed in place" by the action of a peroxidase in the process
described as tyramide signal amplification (TSA). These substrates
are extensively utilized to label targets in samples that are
cells, tissues or arrays for their subsequent detection by
microscopy, flow cytometry, optical scanning and fluorometry.
[0110] Additional colorimetric (and in some cases fluorogenic)
substrate and enzyme combination use a phosphatase enzyme such as
an acid phosphatase, an alkaline phosphatase or a recombinant
version of such a phosphatase in combination with a colorimetric
substrate such as 5-bromo-6-chloro-3-indolyl phosphate (BCIP),
6-chloro-3-indolyl phosphate, 5-bromo-6-chloro-3-indolyl phosphate,
p-nitrophenyl phosphate, or o-nitrophenyl phosphate or with a
fluorogenic substrate such as 4-methylumbelliferyl phosphate,
6,8-difluoro-7-hydroxy4-methylcoumarinyl phosphate (DiFMUP)
fluorescein diphosphate, 3-O-methylfluorescein phosphate, resorufin
phosphate, 9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl)
phosphate (DDAO phosphate), or ELF 97, ELF 39 or related
phosphates.
[0111] Glycosidases, in particular .beta.-galactosidase,
.beta.-glucuronidase and .beta.-glucosidase, are additional
suitable enzymes. Appropriate colorimetric substrates include, but
are not limited to, 5-bromo4-chloro-3-indolyl
.beta.-D-galactopyranoside (X-gal) and similar indolyl
galactosides, glucosides, and glucuronides, o-nitrophenyl
.beta.-D-galactopyranoside (ONPG) and p-nitrophenyl
.beta.-D-galactopyranoside. Preferred fluorogenic substrates
include resorufin .beta.-D-galactopyranoside, fluorescein
digalactoside (FUG), fluorescein diglucuronide and their structural
variants, 4-methylumbelliferyl .beta.-D-galactopyranoside,
carboxyumbelliferyl .beta.-D-galactopyranoside and fluorinated
coumarin .beta.-D-galactopyranosides. Additional enzymes
includehydrolases such as cholinesterases and peptidases, oxidases
such as glucose oxidase and cytochrome oxidases, and reductases for
which suitable substrates are known.
[0112] Enzymes and their appropriate substrates that produce
chemiluminescence are preferred for some assays. These include, but
are not limited to, natural and recombinant forms of luciferases
and aequorins. Chemiluminescence-producing substrates for
phosphatases, glycosidases and oxidases such as those containing
stable dioxetanes, luminol, isoluminol and acridinium esters are
additionally useful. For example, the enzyme is luciferase or
aequorin. The substrates are luciferin, ATP, Ca.sup.++ and
coelenterazine.
[0113] In addition to enzymes, haptens such as biotin are useful
detectable moieties. Biotin is useful as it is in an enzyme system
that can further amplify a detectable signal, and it can serve as a
tag in affinity chromatography for isolation purposes. For
detection, an enzyme conjugate that has affinity for biotin is
used, such as avidin-HRP. Subsequently, a peroxidase substrate is
added to produce a detectable signal. Haptens also include
hormones, naturally occurring and synthetic drugs, pollutants,
allergens, affector molecules, growth factors, chemokines,
cytokines, lymphokines, amino acids, peptides, chemical
intermediates, or nucleotides.
[0114] In some cases, a detectable moiety is a fluorescent protein.
Exemplary fluorescent proteins include green fluorescent protein
(GFP), phycobiliproteins and their derivatives, luciferase or
aequorin. Fluorescent proteins, especially phycobiliprotein, are
particularly useful for creating tandem dye labeled labeling
reagents. These tandem dyes comprise a fluorescent protein and a
fluorophore to obtain a larger stokes shift where the emission
spectra is farther shifted from the fluorescent protein's
absorption spectra. This is particularly advantageous to detect a
low amount of target in a sample where the emitted fluorescent
light is maximally optimized, in other words the fluorescent
protein reabsorbs little to none of the emitted light. The
fluorescent protein and fluorophore function as an energy transfer
pair where the fluorescent protein emits at a wavelength the
fluorophore absorbs, and the fluorphore then emits at a wavelength
farther from the fluorescent protein than could be obtained with
only the fluorescent protein. A particularly useful combination is
phycobiliproteins and sulforhodamine fluorophores, or sulfonated
cyanine fluorophores; or sulfonated xanthene derivatives.
Alternatively, the fluorophore is an energy donor and the
fluorescent protein is an energy acceptor.
Methods of Visualizing the Detection Moiety Depend on the
Label.
[0115] In some cases, a sample is illuminated with a light
wavelength selected to give a detectable optical response, and
observed with a means for detecting that response. Equipment useful
for illuminating fluorescent compounds include hand-held
ultraviolet lamps, mercury arc lamps, xenon lamps, lasers and laser
diodes. These illumination sources are optically integrated into
laser scanners, fluorescent microplate readers or standard or
microfluorometers. The degree or location of signal, compared to a
standard or expected response, indicates whether and to what degree
the sample possesses a given characteristic or desired target.
[0116] An optical response is detected by visual inspection, or by
using one of the following devices: CCD camera, video camera,
photographic film, laser-scanning devices, fluorometers,
photodiodes, quantum counters, epifluorescence microscopes,
scanning microscopes, flow cytometers, fluorescence microplate
readers, or by means for amplifying the signal such as
photomultiplier tubes. When a sample is examined using a flow
cytometer, examination of it optionally includes sorting portions
of it according to their fluorescence response.
[0117] When an indirectly detectable label is used, then
illuminating typically includes adding a reagent to produce a
detectable signal such as a colorimetric enzyme substrate.
Radioisotopes are also considered indirectly detectable where an
additional reagent is not needed, rather the radioisotope is
exposed to X-ray film or other mechanism to record and measure the
signal. This is true for some chemiluminescent signals that are
observed after exposure to film.
I. ONC201 (COMPOUND (1)), SALTS THEREOF AND SYNTHESES THEREOF
[0118] ONC201 (compound (1))
##STR00003##
and its analogs, and their pharmaceutically acceptable salts, as
well as syntheses for them, are provided herein. In in vitro
models, animal models, and human clinical trials, ONC201 has broad
anti-cancer activity, low toxicity including few, if any, adverse
effects, low genotoxicity, and high bioavailability including
orally. These features allow ONC201 and various analogs to be well
suited for a variety of applications. ONC201 can be made by the
synthesis shown in Scheme 1.
##STR00004##
[0119] Synthesis of an ONC201 dihydrochloride salt starts with
commercially available intermediary
N-Benzyl-3-carbomethoxy-4-piperidone hydrochloride, compound (3).
In one embodiment, the synthesis includes neutralizing compound (3)
with a base (Step 1) to produce compound (4), a free base. In one
embodiment, compound (3) is neutralized with an inorganic base to
produce compound (4). In one embodiment, compound (3) is
neutralized with an organic base to produce compound (4). In one
embodiment, compound (3) is neutralized in the presence of an
alcohol, for example, n-butanol. In one embodiment, compound (3) is
neutralized in the presence of at least one organic solvent, for
example, n-butanol and/or ethyl acetate. In one embodiment,
compound (3) is neutralized in the presence of a base and at least
one organic solvent, for example, NaHCO.sub.3 and n-butanol. In one
embodiment, compound (3) is neutralized in the presence of
n-butanol and triethyl amine (Et.sub.3N).
[0120] In one embodiment, the synthesis includes reacting compound
(4) with compound (5) (Step 2) to produce intermediary compound
(1). In one embodiment, the reaction in Step 2 includes heating
compound (4) with compound (5). In one embodiment, the reaction in
Step 2 includes refluxing heating compound (4) and compound (5) in
the presence of a solvent. In one embodiment, the reaction in Step
2 includes use of Dean-stark trap to remove water and/or methanol
(MeOH) formed in the reaction.
[0121] In one embodiment, an ONC201 dihydrochloride salt is
synthesized (Step 3). In one embodiment, this reaction (Step 3)
includes treating ONC201 with HCl in dioxane. In one embodiment,
Step 3 includes treating ONC201 with 4N HCl in dioxane. In one
embodiment, the synthesis optionally includes recrystallizing the
ONC201 di-salt. In a preferred embodiment, the ONC201
di-hydrochloride salt is synthesized as shown in Scheme 2.
##STR00005##
II. TNF-RELATED APOPTOSIS-INDUCING LIGAND ("TRAIL")
[0122] TRAIL protein can be assayed in a sample obtained from a
subject to detect TRAIL expression induced by compounds and their
salts described herein Immunoassays can be used to assay TRAIL in a
sample, including enzyme-linked immunosorbent assay (ELISA),
enzyme-linked immunofiltration assay (ELIFA), flow cytometry,
immunoblot, immunoprecipitation, immunohistochemistry,
immunocytochemistry, luminescent immunoassay (LIA), fluorescent
immunoassay (FIA), and radioimmunoassay. Assays may be used to
obtain qualitative and/or quantitative results. Specific details of
suitable methods for both qualitative and quantitative sample
assays are described in standard references, including E. Harlow
& D. Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor
Laboratory Press, 1988; F. Breitling & S. 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; B. K. C. Lo, Antibody
Engineering: Methods and Protocols, Methods in Molecular Biology,
Humana Press, 2003; F. M. Ausubel et al., Eds., Short Protocols in
Molecular Biology, Current Protocols, Wiley, 2002; S. Klussman,
Ed., The Aptamer Handbook: Functional Oligonucleotides and Their
Applications, Wiley, 2006; Ormerod, M. G., Flow Cytometry: a
practical approach, Oxford University Press, 2000; Givan, A. L.,
Flow Cytometry: first principles, Wiley, New York, 2001; Gorczyca,
W., Flow Cytometry in Neoplastic Hematology:
morphologic-immunophenotypic correlation, Taylor & Francis,
2006; Crowther, J. R., The ELISA Guidebook (Methods in Molecular
Biology), Humana Press, 2000; Wild, D., The Immunoassay Handbook,
3.sup.rd Edition, Elsevier Science, 2005, and J. Sambrook and D. W.
Russell, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor
Laboratory Press, 3.sup.rd ed., 2001.
[0123] Protocols to assay and analyze a sample for TRAIL to detect
an effect of a pharmaceutical composition are described in U.S.
Pat. No. 8,673,923 to Wafik S. El-deiry et al., which is
incorporated by reference herein in its entirety.
[0124] In one embodiment, TRAIL assays are used to monitor a
subject. For example, a sample is obtained from a subject before
treatment with a pharmaceutical composition and at one or more
times during and/or following treatment to assess the treatment's
effectiveness. In another example, a sample is obtained from a
subject at various times to assess the course or progress of
disease or healing. In one embodiment, death receptors from
circulating tumor cells are assayed to see if a treatment described
here increases the amount or type of death receptors.
[0125] Cancers treated using methods and compositions described
herein are characterized by abnormal cell proliferation including
pre-neoplastic hyperproliferation, cancer in-situ, neoplasms and
metastasis. Methods and compositions described herein can be used
for prophylaxis, as well as amelioration of cancer signs or
symptoms. "Treatment" of a cancer in a subject includes:
preventing, inhibiting or ameliorating cancer in the subject, such
as slowing cancer progression or reducing or ameliorating a cancer
sign or symptom. Examples of cancers treated using methods and
compositions described herein include breast cancer, CNS cancers,
colon cancer, ovarian cancer, prostate cancer, leukemia, lung
cancer, and lymphoma.
III. COMPOUNDS OF FORMULA (10) AND SALTS THEREOF
[0126] In one aspect, provided herein are compounds and salts of
formula (10) and methods of making them. Persons skilled in the art
will understand that the general principles and concepts described
here in conjunction with ONC201 (compound (1)) and its salts,
including principles and concepts related to methods and
pharmaceutical compositions, apply with equal force to compounds of
formula (10) and salts thereof.
[0127] In one embodiment, provided herein are compounds of formula
(10):
##STR00006##
wherein R.sub.1 and R.sub.2 are independently selected from H,
alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl,
heterocycloalkylalkyl, heteroaryl, arylalkyl, heteroarylalkyl,
alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio, and acyl
radicals. In one embodiment, R.sub.1 is CH.sub.2Ph and R.sub.2 is
CH.sub.2-(2-CH.sub.3-Ph) (i.e., ONC201). In one embodiment, R.sub.1
is CH.sub.2Ph and R.sub.2 is CH.sub.2-(2,4-di F-Ph) (i.e., ONC206).
In one embodiment, R.sub.1 is CH.sub.2Ph and R.sub.2 is
CH.sub.2-(4-CF.sub.3-Ph) (i.e., ONC212). In one embodiment, R.sub.1
is CH.sub.2Ph and R.sub.2 is CH.sub.2-(3,4-di F-Ph) (i.e., ONC213).
In one embodiment, R.sub.1 is CH.sub.2 (3,4-di-Cl-Ph and R.sub.2 is
CH.sub.2-(4-CF.sub.3-Ph) (i.e., ONC234). In one embodiment, R.sub.1
is CH.sub.2-3-thienyl and R.sub.2 is CH.sub.2-(4-CF.sub.3-Ph)
(i.e., ONC236).
[0128] In one embodiment, R.sub.1 and R.sub.2 are independently
selected from the group consisting of H, C.sub.1-4alkyl,
C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
C.sub.1-4benzyl-piperazine, C.sub.1-4alkylthienyl,
C.sub.1-4alkylpyridinyl, C.sub.1-4alkylisoxazolidinyl,
C.sub.1-4alkylmorpholinyl, C.sub.1-4alkylthiazolyl, and
C.sub.1-4alkylpyrazinyl wherein C.sub.1-4alkyl,
C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone, C.sub.1-4
benzyl-piperazine, C.sub.1-4alkylthienyl, C.sub.1-4alkylpyridinyl,
C.sub.1-4alkylisoxazolidinyl, C.sub.1-4alkylmorpholinyl,
C.sub.1-4alkylthiazolyl, and C.sub.1-4alkylpyrazinyl are optionally
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo. In one embodiment, R.sub.1
and/or R.sub.2 is a substituted or unsubstituted, arylalkyl or
heteroarylalkyl. In one embodiment, the heteroarylalkyl is selected
from C.sub.1-4alkylpyrrolyl, C.sub.1-4alkylfuryl,
C.sub.1-4alkylpyridyl, C.sub.1-4alkyl-1,2,4-thiadiazolyl,
C.sub.1-4alkylpyrimidyl, C.sub.1-4alkylthienyl,
C.sub.1-4alkylisothiazolyl, C.sub.1-4alkylimidazolyl,
C.sub.1-4alkyltetrazolyl, C.sub.1-4alkylpyrazinyl,
C.sub.1-4alkylpyrimidyl, C.sub.1-4alkylquinolyl,
C.sub.1-4alkylisoquinolyl, C.sub.1-4alkylthiophenyl,
C.sub.1-4alkylbenzothienyl, C.sub.1-4alkylisobenzofuryl,
C.sub.1-4alkylpyrazolyl, C.sub.1-4alkylindolyl,
C.sub.1-4alkylpurinyl, C.sub.1-4alkylcarbazolyl,
C.sub.1-4alkylbenzimidazolyl, and C.sub.1-4alkylisoxazolyl.
[0129] In one embodiment, R.sub.1 and/or R.sub.2 is a benzyl
optionally substituted with one or more of the following
substituents on the benzyl ring: X, --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CN, --CXH.sub.2, --CX.sub.2H, C.sub.2-C.sub.4 alkyl,
--CX.sub.3, --CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2,
--C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1, --OCX.sub.3,
--OC.sub.pH.sub.2p+1, --OC.sub.pX.sub.2p+1, OR.sup.m, SR.sup.m,
NR.sup.mR.sup.n, NR.sup.mC(O)R.sup.n, SOR.sup.m, SO.sub.2R.sup.m,
C(O)R.sup.m, and C(O)OR.sup.m; R.sup.m and R.sup.n are
independently selected from H or a C.sub.1-C.sub.4 alkyl; and where
p is an integer from 2 to 20 and X is a halogen, including F, Cl,
Br, or I; preferably, F, Cl, or Br; more preferably, F or Cl.
[0130] In one embodiment, R.sub.1 is selected from H, CH.sub.3,
CH.sub.2Ph, CH.sub.2-(4-CF.sub.3-Ph), CH.sub.2-(4-F-Ph),
CH.sub.2-(4-Cl-Ph), CH.sub.2--(OCH.sub.3-Ph), CH.sub.2-((2-Cl)-Ph),
CH.sub.2-(2-thienyl), CH.sub.2-(3-thienyl), CH.sub.2-2-pyridinyl,
CH.sub.2-4-methyl-2-thiazolyl, CH.sub.2-2-pyrazinyl,
CH.sub.2CH.sub.2Ph, CH.sub.2CH.sub.2(4-N-benzyl-piperazine),
CH.sub.2-(2,4-di F-Ph), CH.sub.2-(3,4-di Cl-Ph), CH.sub.2-(3,4-di
F-Ph), CH.sub.2-(3,5-di F-Ph), CH.sub.2-((2-CH.sub.3)-Ph),
CH.sub.2CH(OH)Ph, (4-F-Ph)-4-oxobutyl,
CH.sub.2CH.sub.2NHCOOC(CH.sub.3).sub.3,
CH.sub.2CH.sub.2CH.sub.2NH.sub.2, and CD.sub.2C.sub.6D.sub.5. In
one embodiment, R.sub.2 is selected from H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-(4-CF.sub.3-Ph), CH.sub.2-((2-Cl)-Ph),
CH.sub.2-((2-F)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-(2,4-di Cl-Ph), CH.sub.2-(3,4-di Cl-Ph), CH.sub.2-(3,4-di
F-Ph), CH.sub.2-(3,5-di F-Ph), CH.sub.2-((2-CH.sub.3)-Ph),
CH.sub.2(2-CH.sub.3, 4-F-Ph), CH.sub.2-((4-OCH.sub.3)-Ph),
CH.sub.2-(3-pyridinyl), CH.sub.2-(3-isoxazolidinyl),
CH.sub.2CH.sub.2-(4-morpholinyl), CH.sub.2-(2-F, 4-CF.sub.3-Ph),
CH.sub.2CH(OH)Ph, (CH.sub.2).sub.3CO-4F-Ph, (4-F-Ph)-4-oxobutyl,
CH.sub.2CH.sub.2NHCOOC(CH.sub.3).sub.3,
CH.sub.2CH.sub.2CH.sub.2NH.sub.2, and CD.sub.2C.sub.6D.sub.5.
[0131] In one embodiment, R.sub.1 is H. In one embodiment, R.sub.1
is a substituted or unsubstituted arylalkyl, e.g., a benzyl
(CH.sub.2Ph) or phenylethyl group. In one embodiment, the arylalkyl
is substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo.
[0132] In one embodiment, R.sub.2 is a substituted or an
unsubstituted arylalkyl, e.g., benzyl or phenylethyl. In one
embodiment, the arylalkyl is substituted with C.sub.1-4alkyl,
C.sub.1-4alkoxyl, hydroxyl, perhalogenated C.sub.1-4alkyl, or halo.
In one embodiment, the arylalkyl is substituted with one or more
substituents selected from halo, CH.sub.3, CF.sub.3 or OCH.sub.3.
In one embodiment, R.sub.2 is a substituted or an unsubstituted
heterocycloalkylalkyl, e.g., piperazinylalkyl or morpholinoalkyl.
In one embodiment, R.sub.2 is a substituted or an unsubstituted
heteroarylalkyl, e.g., pyridylmethyl or isoxazolidinylmethyl. In
one embodiment, the heterocycloalkylalkyl or heteroarylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo. In one embodiment, the
heterocycloalkylalkyl or heteroarylalkyl is substituted with at
least one substituent selected from halo, CH.sub.3, CF.sub.3 or
OCH.sub.3.
[0133] In one embodiment, compound (10) has the structure of
formula (80):
##STR00007##
wherein R.sub.a1, R.sub.a2, R.sub.a3, R.sub.a4, R.sub.a5, R.sub.b1,
R.sub.b2, R.sub.b3, R.sub.b4, and R.sub.b5 are each independently
selected from the group consisting of H, X, --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CN, --CXH.sub.2, --CX.sub.2H, C.sub.2-C.sub.4 alkyl,
--CX.sub.3, --CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2,
--C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1, --OCX.sub.3,
--OC.sub.pH.sub.2p+1, --OC.sub.pX.sub.2p+1, OR.sup.m, SR.sup.m,
NR.sup.mR.sup.n, NR.sup.mC(O)R.sup.n, SOR.sup.m, SO.sub.2R.sup.m,
C(O)R.sup.m, and C(O)OR.sup.m; R.sup.m and R.sup.n are
independently selected from H or a C.sub.1-C.sub.4 alkyl; and where
p is an integer from 2 to 20 and X is a halogen.
[0134] In one embodiment, compound (10) has the structure of
formula (90)
##STR00008##
wherein R.sub.2 is as defined above, and wherein R.sub.b1,
R.sub.b2, R.sub.b3, R.sub.b4, and R.sub.b5 are each independently
selected from the group consisting of H, X, --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CN, --CXH.sub.2, --CX.sub.2H, C.sub.2-4 alkyl,
--CX.sub.3, --CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2,
--C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1, --OCX.sub.3,
--OC.sub.pH.sub.2p+1, --OC.sub.pX.sub.2p+1, OR.sup.m, SR.sup.m,
NR.sup.mR.sup.n, NR.sup.mC(O)R.sup.n, SOR.sup.m, SO.sub.2R.sup.m,
C(O)R.sup.m, and C(O)OR.sup.m; R.sup.m and R.sup.n are
independently selected from H or a C.sub.1-4alkyl; and where p is
an integer from 2 to 20 and X is a halogen.
[0135] In one embodiment, compound (10) has the structure of
formula (40)
##STR00009##
where R.sub.1 is as defined above, and where R.sub.a1, R.sub.a2,
R.sub.a3, R.sub.a4, and R.sub.a5 are each independently selected
from the group consisting of H, X, --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CN, --CXH.sub.2, --CX.sub.2H, C.sub.2-4 alkyl,
--CX.sub.3, --CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2,
--C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1, --OCX.sub.3,
--OC.sub.pH.sub.2p+1, --OC.sub.pX.sub.2p+1, OR.sup.m, SR.sup.m,
NR.sup.mR.sup.n, NR.sup.mC(O)R.sup.n, SOR.sup.m, SO.sub.2R.sup.m,
C(O)R.sup.m, and C(O)OR.sup.m; R.sup.m and R.sup.n are
independently selected from H or a C.sub.1-4 alkyl; p is an integer
from 2 to 20; and X is a halogen. In one embodiment, R.sub.1 is H.
In one embodiment, R.sub.1 is a substituted or unsubstituted
arylalkyl, such as benzyl or phenylethyl. In one embodiment, the
arylalkyl is substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl,
hydroxyl, perhalogenated C.sub.1-4alkyl, or halo. In one
embodiment, the benzyl is substituted with one or more halo. In one
embodiment, the benzyl is substituted with one or more substituents
selected from halo, CH.sub.3, CF.sub.3, and OCH.sub.3. In one
embodiment, the benzyl is substituted with one halo, e.g., F at an
ortho or para position. In one embodiment, the benzyl is
substituted with two halogen, e.g., F at both meta positions.
[0136] In one embodiment, compound (40) has the structure of
compound (45):
##STR00010##
where R.sub.a1, R.sub.a2, R.sub.a3, R.sub.a4, and R.sub.a5 are as
defined above. In one embodiment, the benzyl is substituted with
one or more halogens. In one embodiment, the benzyl is substituted
with one or more substituents selected from halo, CH.sub.3,
CF.sub.3, and OCH.sub.3. In one embodiment, R.sub.a1 or R.sub.a5 is
a halo, e.g., F. In one embodiment, both R.sub.a2 and R.sub.a3 are
halo, e.g., F.
[0137] In one embodiment, compound (10) has the structure of
compound (50)
##STR00011##
wherein R.sub.1 is as defined above, and wherein Rb is selected
from the group consisting of H, X, --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CN, --CXH.sub.2, --CX.sub.2H, C.sub.2-4alkyl,
--CX.sub.3, --CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2,
--C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1, --OCX.sub.3,
--OC.sub.pH.sub.2p+1, --OC.sub.pX.sub.2p+1, OR.sup.m, SR.sup.m,
NR.sup.mR.sup.n, NR.sup.mC(O)R.sup.n, SOR.sup.m, SO.sub.2R.sup.m,
C(O)R.sup.m, and C(O)OR.sup.m; R.sup.m and R.sup.n are
independently selected from H or C.sub.1-4alkyl; and where p is an
integer from 2 to 20 and X is a halogen, and wherein R.sub.a1,
R.sub.a2, R.sub.a4, and R.sub.a5 are each independently selected
from the group consisting of H, X, --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CN, --CXH.sub.2, --CX.sub.2H, C.sub.2-4alkyl,
--CX.sub.3, --CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2,
--C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1, --OCX.sub.3,
--OC.sub.pH.sub.2p+1, --OC.sub.pX.sub.2p+1, OR.sup.m, SR.sup.m,
NR.sup.mR.sup.n, NR.sup.mC(O)R.sup.n, SOR.sup.m, SO.sub.2R.sup.m,
C(O)R.sup.m, and C(O)OR.sup.m; R.sup.m and R.sup.n are
independently selected from H or C.sub.1-4 alkyl; and where p is an
integer from 2 to 20 and X is a halogen. In one embodiment, R.sub.1
is H. In one embodiment, R.sub.1 is a substituted or unsubstituted
arylalkyl, such as a benzyl or phenylethyl group. In one
embodiment, the arylalkyl is substituted with C.sub.1-4alkyl,
C.sub.1-4alkoxyl, hydroxyl, perhalogenated C.sub.1-4alkyl, or halo.
In one embodiment, Rb is selected from halo, CH.sub.3, CF.sub.3,
and OCH.sub.3. In one embodiment, one or more of R.sub.a1,
R.sub.a2, R.sub.a4, and R.sub.a5 is selected from halo, CH.sub.3,
CF.sub.3, and OCH.sub.3. In one embodiment, R.sub.a1, R.sub.a2,
R.sub.a4, and R.sub.a5 are H, and R.sub.b is selected from halo,
CH.sub.3, CF.sub.3, and OCH.sub.3. In one embodiment, R.sub.b is a
halogen, e.g., F, and R.sub.a1 is CH.sub.3. In one embodiment,
R.sub.b is F or Cl, and R.sub.a2 is F or Cl. In one embodiment,
R.sub.b is CF.sub.3. In one embodiment, R.sub.b is OCH.sub.3. In
one embodiment, R.sub.b and R.sub.a1 are Cl.
[0138] In one embodiment, compound (50) has the structure of
compound (55):
##STR00012##
where R.sub.a1, R.sub.a2, R.sub.a4, R.sub.a5, and R.sub.b are as
defined above. In one embodiment, R.sub.b is selected from halo,
CH.sub.3, CF.sub.3, and OCH.sub.3. In one embodiment, one or more
of R.sub.a1, R.sub.a2, R.sub.a4, and R.sub.a5 is selected from
halo, CH.sub.3, CF.sub.3, and OCH.sub.3. In one embodiment,
R.sub.a1, R.sub.a2, R.sub.a4, and R.sub.a5 are H, and R.sub.b is
selected from halo, CH.sub.3, CF.sub.3, and OCH.sub.3. In one
embodiment, R.sub.b is halo, e.g., F, and R.sub.a1 is CH.sub.3. In
one embodiment, R.sub.b is F or Cl, and R.sub.a2 is F or Cl. In one
embodiment, R.sub.b is CF.sub.3. In one embodiment, R.sub.b is
OCH.sub.3. In one embodiment, R.sub.b and R.sub.a1 are C.sub.1.
[0139] In one embodiment, compound (10) has the structure of
compound (60)
##STR00013##
In one embodiment, R.sub.1 is H. In one embodiment, R.sub.1 is a
substituted or unsubstituted arylalkyl, such as benzyl or
phenylethyl. In one embodiment, R.sub.1 is a substituted or
unsubstituted heterocycloalkylalkyl or a substituted or
unsubstituted heteroarylalkyl, such as CH.sub.2-(2-thienyl),
CH.sub.2-(3-thienyl), CH.sub.2-4-methyl-2-thiazolyl,
CH.sub.2-2-pyrazinyl, CH.sub.2CH.sub.2(4-N-benzyl-piperazine),
CH.sub.2-(3-isoxazolidinyl), CH.sub.2-2-pyridinyl,
CH.sub.2-3-pyridinyl, and CH.sub.2CH.sub.2-(4-morpholinyl). In one
embodiment, the arylalkyl is substituted with C.sub.1-4alkyl,
C.sub.1-4alkoxyl, hydroxyl, perhalogenated C.sub.1-4alkyl, or halo.
In one embodiment, the benzyl is substituted with one or more
halogens. In one embodiment, the benzyl is substituted with one or
more substituents selected from halo (e.g., F), CH.sub.3, CF.sub.3,
and OCH.sub.3. In one embodiment, the benzyl is substituted at the
para position with a halo, CH.sub.3, CF.sub.3, or OCH.sub.3
substituent. In one embodiment, R.sub.1 is fluorophenyloxobutyl or
hydroxyphenylethyl Scheme 3 illustrates the synthesis of compounds
of formula (10):
##STR00014##
[0140] Compounds of formula (10) (i.e., imipridones) are
synthesized starting from a substituted piperidone, which is
converted by reaction with a substituted aminoimidazoline to give
the core compound (10). There are two routes, one in which the
R.sub.1 substituent is present in the piperidone (e.g., 68). In
that route, (68) is acylated with dimethyl carbonate using sodium
hydride in toluene at 80.degree. C. to form piperidone ester (69).
Commercially available methylthioimidazoline HI salt (63) is
reacted with an amine in dioxane at 70.degree. C. to afford the
R.sub.2-substituted aminoimidazoline (64) as its HI salt. Direct
reaction of (64) with piperidone ester (69) in 1-butanol at reflux
with removal of water via a Dean-Stark trap over 3-6 h gives the
tricylic compound (10). In a variant of this scheme, N--BOC
protected piperidone (61) is converted by the same methods to BOC
protected compound (65), which is treated with HCl in dioxane to
remove the BOC group and then converted to the free base of (66)
with 1N NaOH with extraction with methylene chloride. Subsequent
treatment of (66) with a halide (67) or epoxide (70) affords
desired compound (10).
[0141] Crude products may be purified by column chromatography
eluting with methylene chloride:methanol or by HPLC using
acetonitrile:TFA:H.sub.2O to produce final products as either free
bases or as TFA salts. Treatment of free bases with HCl in dioxane
or lyophilization of TFA salts generates products (10) as HCl or
TFA salts. Alternatively, the free base may be treated with another
inorganic or organic acid to form other salts, generally selected
from those known to be pharmaceutically acceptable. Salts of
compound (10) are usually solids and examples have been
crystallized from ethanol or other solvents to give high quality
crystals. The tricyclic structure has been definitively confirmed
in the case of compound (1) by an X-ray crystal structure and
NMR.
[0142] Compounds described herein can be used, with or without an
aminoalkyl linker (e.g., compound (33)), to identify molecules
(e.g., proteins) that interact with them in a cellular context.
Expression of these binding targets may be used to predict response
to imipridones or analogs thereof (i.e. serve as biomarkers). These
compounds can also be used to screen for structurally unrelated
molecules using competition assays known in the art to identify
drugs able to outcompete the target interaction with a higher
affinity. In addition, these molecules may have improved drug
properties or allow additional applications by altering drug
properties including safety, potency, pharmacokinetics,
biodistribution, or metabolism.
TABLE-US-00001 TABLE 1 EXAMPLES OF COMPOUNDS OF FORMULA (10) No.
ONC Number R.sub.1 R.sub.2 1 ONC201 CH.sub.2Ph
CH.sub.2-((2-CH.sub.3)--Ph) 13 CH.sub.2Ph CH.sub.3 14 ONC202
CH.sub.2Ph CH.sub.2-((2-Cl)--Ph) 15 ONC203 CH.sub.2Ph
CH.sub.2-(2-thienyl) 16 ONC204 CH.sub.2Ph CH.sub.2CH.sub.2Ph 17
ONC205 CH.sub.2Ph CH.sub.2CH.sub.2(4-N- benzyl-piperazine) 18
ONC206 CH.sub.2Ph CH.sub.2-(2,4-di F--Ph) 19 ONC207 H
CH.sub.2-((2-CH.sub.3)--Ph) 20 ONC208 CH.sub.3
CH.sub.2-((2-CH.sub.3)--Ph) 21 ONC209 CH.sub.2CH.sub.2Ph
CH.sub.2-((2-CH.sub.3)--Ph) 22 CH.sub.2CH.sub.2-(4-N-
CH.sub.2-((2-CH.sub.3)--Ph) benzyl-piperizine) 23 CH.sub.2CHOHPh
CH.sub.2-((2-CH.sub.3)--Ph) 24 (CH.sub.2).sub.3CO--4F--Ph
CH.sub.2-((2-CH.sub.3)--Ph) 32 ONC215
CH.sub.2CH.sub.2NHCOOC(CH.sub.3).sub.3 CH.sub.2-((2-CH.sub.3)--Ph)
33 ONC216 CH.sub.2CH.sub.2CH.sub.2NH.sub.2
CH.sub.2-((2-CH.sub.3)--Ph) 41 ONC210 CH.sub.2Ph CH.sub.2-(3,5-di
F--Ph) 51 ONC211 CH.sub.2Ph CH.sub.2-(3,4-di Cl--Ph) 52 ONC212
CH.sub.2Ph CH.sub.2-(4-CF.sub.3--Ph) 53 ONC213 CH.sub.2Ph
CH.sub.2-(3,4-di F--Ph) 54 ONC214 CD.sub.2C.sub.6D.sub.5
CH.sub.2-((2-CH.sub.3)--Ph) 43 ONC217 CH.sub.2Ph CH.sub.2-(2-F--Ph)
55 ONC218 CH.sub.2Ph CH.sub.2(2-CH.sub.3, 4-F--Ph) 56 ONC219
CH.sub.2Ph CH.sub.2-(2,4-di Cl--Ph) 57 ONC220 CH.sub.2Ph
CH.sub.2-((4-OCH.sub.3)--Ph) 34 ONC226 CH.sub.2Ph
CH.sub.2-(3-pyridinyl) 35 ONC222 CH.sub.2Ph
CH.sub.2-(3-isoxazoliclinyl) 36 ONC224 CH.sub.2Ph
CH.sub.2CH.sub.2-(4-morpholinyl) 37 ONC223 CH.sub.2Ph
CH.sub.2-(4-CH.sub.3--Ph) 38 ONC221 H CH.sub.2-(4-CF.sub.3--Ph) 73
ONC227 CH.sub.2-(4-CF.sub.3--Ph) CH.sub.2-(4-CF.sub.3--Ph) 72
ONC225 CH.sub.2Ph CH.sub.2-(2-F, 4-CF.sub.3--Ph) 74 ONC228
CH.sub.2-(4-F--Ph) CH.sub.2-(4-CF.sub.3--Ph) 75 ONC229
CH.sub.2-(OCH.sub.3--Ph) CH.sub.2-(4-CF.sub.3--Ph) 76 ONC230
(4-F--Ph)-4-oxobutyl CH.sub.2-(4-CF.sub.3--Ph) 77 ONC231
CH.sub.2-3-pyridyl CH.sub.2-(4-CF.sub.3--Ph) 78 ONC232
CH.sub.2-4-methyl-2-thiazolyl CH.sub.2-(4-CF.sub.3--Ph) 79 ONC233
CH.sub.2-2-pyrazinyl CH.sub.2-(4-CF.sub.3--Ph) 81 ONC234
CH.sub.2-(3,4-di Cl--Ph) CH.sub.2-(4-CF.sub.3--Ph) 82 ONC235
CH.sub.2-(4-Cl--Ph) CH.sub.2-(4-CF.sub.3--Ph) 83 ONC236
CH.sub.2-3-thienyl CH.sub.2-(4-CF.sub.3--Ph) 84 ONC237
CH.sub.2CH(OH)Ph CH.sub.2-(4-CF.sub.3--Ph)
IV. ASSESSING SENSITIVITY AND EFFICACY OF TREATMENT REGIMENS
[0143] Measuring expression, gene mutation, or gene copy number of
a dopamine receptor or another G protein-coupled receptor (GPCR)
may be used to predict response or sensitivity to a method of
treatment described herein and to identify subjects likely to be
responsive to a method of treatment described herein, such as
treatment with a compound of formula (10), a pharmaceutically
acceptable salt thereof, or an analog thereof. In one aspect,
provided herein are methods of identifying whether a subject having
a condition is likely to be responsive to a treatment regimen
described herein. In one embodiment, the methods comprises (i)
obtaining a biological sample from the subject; (ii) measuring
expression levels of at least one dopamine receptor or G
protein-coupled receptor (GPCR) in the sample; (iii) comparing the
levels measured in the sample to those for a pre-determined
standard; and (iv) determining whether the subject is likely to be
responsive to the treatment regimen, based on the levels measured
in the sample to those for the pre-determined standard. In one
embodiment, the step of measuring an expression level of a dopamine
receptor or GPCR in the sample include the steps of (i) contacting
the sample with an antibody or antigen-binding fragment that
specifically binds to the receptor to form a complex of the
antibody or antigen-binding fragment with the receptor; and (ii)
measuring the amount of the complex. In one embodiment, the subject
has, or is at risk of having, cancer. In one embodiment, the cancer
is a neuro-oncology disease. In one embodiment, the cancer is a
neuroendocrine tumor. In one embodiment, the cancer is selected
from the group consisting of meningioma, ependymoma, glioma,
neuroblastoma, and diffuse intrinsic pontine glioma. In one
embodiment, the subject has, or is at risk of having, a psychiatric
disorder. In one embodiment, the psychiatric disorder is selected
from the group consisting of psychosis, bipolar disorder, and major
depressive disorder. In one embodiment, the subject has, or is at
risk of having, an infection. In one embodiment, the infection is a
bacterial infection. In one embodiment, the infection is a
gram-negative bacterial infection. In one embodiment, the infection
is a gram-positive bacterial infection. In one embodiment, the
bacterial infection is an infection of a bacteria selected from the
group consisting of Enterococcus faecium, Staphylococcus aureus,
Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas
aeruginosa, and Enterobacter species. In one embodiment, the
gram-positive bacterial infection is a Staphylococcus infection. In
one embodiment, the Staphylococcus infection is an S. aureus
infection (e.g., a methicillin-resistant S. aureus (MRSA)
infection). In one embodiment, the treatment regimen comprises
administering an effective amount of a therapeutic, such as a
compound of formula (10), a pharmaceutically acceptable salt
thereof, or an analog thereof. In one embodiment, the dopamine
receptor is from the D2-like family of dopamine receptors In one
embodiment, the dopamine receptor is DRD2. In one embodiment, the
dopamine receptor is DRD3. In one embodiment, the dopamine receptor
is DRD4. In one embodiment, the dopamine receptor is DRD2, DRD3, or
both. In one embodiment, the GPCR is a Class A GPCR. In one
embodiment, the GPCR is GPR132. In one embodiment, the GPCR is
selected from the group consisting of GPR132, GPR91, MTNR1A,
GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7, and combinations thereof.
In one embodiment, the dopamine receptor is DRD5, the treatment
regimen comprises administering an effective amount of a
therapeutic, such as a compound of formula (10) or a
pharmaceutically acceptable salt thereof, and an increased level of
expression of DRD5 measured in the sample relative to the
pre-determined standard indicates that the subject is or is not
likely to be responsive to the treatment regimen.
[0144] In another aspect, provided herein are methods of assessing
the effectiveness of a treatment regimen described herein,
monitoring, or providing a prognosis for a subject with a
condition. In one embodiment, the methods comprises (i) obtaining a
biological sample from the subject; (ii) measuring expression
levels of at least one dopamine receptor or G protein-coupled
receptor (GPCR) in the sample; (iii) comparing the levels measured
in the sample to those for a pre-determined standard; and (iv)
determining a prognosis or determining whether the subject is
responsive to the treatment regimen, based on the levels measured
in the sample to those for the pre-determined standard. In one
embodiment, the step of measuring an expression level of a dopamine
receptor or GPCR in the sample include the steps of (i) contacting
the sample with an antibody or antigen-binding fragment that
specifically binds to the receptor to form a complex of the
antibody or antigen-binding fragment with the receptor; and (ii)
measuring the amount of the complex. In one embodiment, the methods
comprise (i) obtaining a biological sample from the subject; (ii)
measuring gene copy number or mutations in at least one dopamine
receptor in the sample; (iii) comparing the copy number measured or
mutations found in the sample to those for a pre-determined
standard; and (iv) determining whether the subject is responsive to
the treatment regimen, based on the copy number measured or
mutations found in the sample to those for the pre-determined
standard. In one embodiment, the subject has, or is at risk of
having, cancer. In one embodiment, the cancer is a neuro-oncology
disease. In one embodiment, the cancer is a neuroendocrine tumor.
In one embodiment, the cancer is selected from the group consisting
of meningioma, ependymoma, glioma, neuroblastoma, and diffuse
intrinsic pontine glioma. In one embodiment, the subject has, or is
at risk of having, a psychiatric disorder. In one embodiment, the
psychiatric disorder is selected from the group consisting of
psychosis, bipolar disorder, and major depressive disorder. In one
embodiment, the subject has, or is at risk of having, an infection.
In one embodiment, the infection is a bacterial infection. In one
embodiment, the infection is a gram-negative bacterial infection.
In one embodiment, the infection is a gram-positive bacterial
infection. In one embodiment, the bacterial infection is an
infection of a bacteria selected from the group consisting of
Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae,
Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter
species. In one embodiment, the gram-positive bacterial infection
is a Staphylococcus infection. In one embodiment, the
Staphylococcus infection is an S. aureus infection (e.g., a
methicillin-resistant S. aureus (MRSA) infection). In one
embodiment, the treatment regimen comprises administering an
effective amount of a therapeutic, such as a compound of formula
(10), a pharmaceutically acceptable salt thereof, or an analog
thereof. In one embodiment, the dopamine receptor is selected from
DRD2, DRD2S, DRD2L, and DRD3. In one embodiment, the dopamine
receptor is from the D2-like family of dopamine receptors. In one
embodiment, the dopamine receptor is from the D1-like family of
dopamine receptors. In one embodiment, the dopamine receptor is
DRD1. In one embodiment, the dopamine receptor is DRD2. In one
embodiment, the dopamine receptor is DRD3. In one embodiment, the
dopamine receptor is DRD4. In one embodiment, the dopamine receptor
is DRD5. In one embodiment, the dopamine receptor is DRD2, DRD3, or
both. In one embodiment, the GPCR is a Class A GPCR. In one
embodiment, the GPCR is GPR132. In one embodiment, the GPCR is
selected from the group consisting of GPR132, GPR91, MTNR1A,
GPR162, GPR137, BAI3, LGR4, PTGIR, CXCR7, and combinations
thereof.
[0145] In one embodiment, the dopamine receptor is DRD5, the
treatment regimen comprises administering an effective amount of a
compound of formula (10) or a pharmaceutically acceptable salt
thereof, and an increased level of expression of DRD5 measured in
the sample relative to the pre-determined standard indicates that
the treatment regimen is or is not effective. In one embodiment,
the dopamine receptor is DRD5, the treatment regimen comprises
administering an effective amount of a therapeutic, such as a
compound of formula (10) or a pharmaceutically acceptable salt
thereof, and mutation in the DRD5 gene measured in the sample
indicates that the treatment regimen is or is not effective. In one
embodiment, the dopamine receptor is DRD5, the treatment regimen
comprises administering an effective amount of a therapeutic, such
as a compound of formula (10) or a pharmaceutically acceptable salt
thereof, and the misense mutation Q366R in the DRD5 gene measured
in the sample indicates that the treatment regimen is or is not
effective.
[0146] In another aspect, provided herein are methods of
identifying whether a subject having a condition is likely to be
responsive to a treatment regimen described herein. In one
embodiment, the methods comprises (i) obtaining a biological sample
from the subject; (ii) measuring gene copy number or mutations in
at least one dopamine receptor in the sample; (iii) comparing the
copy number measured or mutations found in the sample to those for
a pre-determined standard; and (iv) determining whether the subject
is likely to be responsive to the treatment regimen, based on the
copy number measured or mutations found in the sample to those for
the pre-determined standard. In one embodiment, the subject has, or
is at risk of having, cancer. In one embodiment, the cancer is a
neuro-oncology disease. In one embodiment, the cancer is a
neuroendocrine tumor. In one embodiment, the cancer is selected
from the group consisting of meningioma, ependymoma, glioma,
neuroblastoma, and diffuse intrinsic pontine glioma. In one
embodiment, the subject has, or is at risk of having, a psychiatric
disorder. In one embodiment, the psychiatric disorder is selected
from the group consisting of psychosis, schizophrenia, bipolar
disorder, and major depressive disorder. In one embodiment, the
subject has, or is at risk of having, an infection. In one
embodiment, the infection is a bacterial infection. In one
embodiment, the infection is a gram-negative bacterial infection.
In one embodiment, the infection is a gram-positive bacterial
infection. In one embodiment, the bacterial infection is an
infection of a bacteria selected from the group consisting of
Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae,
Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter
species. In one embodiment, the gram-positive bacterial infection
is a Staphylococcus infection. In one embodiment, the
Staphylococcus infection is an S. aureus infection (e.g., a
methicillin-resistant S. aureus (MRSA) infection). In one
embodiment, the treatment regimen comprises administering an
effective amount of a therapeutic, such as a compound of formula
(10), a pharmaceutically acceptable salt thereof, or an analog
thereof. In one embodiment, the dopamine receptor is from the
D2-like family of dopamine receptors. In one embodiment, the
dopamine receptor is DRD1. In one embodiment, the dopamine receptor
is DRD2. In one embodiment, the dopamine receptor is DRD3. In one
embodiment, the dopamine receptor is DRD4. In one embodiment, the
dopamine receptor is DRD5. In one embodiment, the dopamine receptor
is DRD2, DRD3, or both. In one embodiment, the dopamine receptor is
DRD5, the treatment regimen comprises administering an effective
amount of a therapeutic, such as a compound of formula (10) or a
pharmaceutically acceptable salt thereof, and mutation in the DRD5
gene measured in the sample indicates that the subject is or is not
likely to be responsive to the treatment regimen. In one
embodiment, the dopamine receptor is DRD5, the treatment regimen
comprises administering an effective amount of a therapeutic, such
as a compound of formula (10) or a pharmaceutically acceptable salt
thereof, and the misense mutation Q366R in the DRD5 gene measured
in the sample indicates that the subject is or is not likely to be
responsive to the treatment regimen.
[0147] In addition, measuring expression, post-translational
modifications, or activity levels of or mutations in eIF2-.alpha.,
ATF4, CHOP, DR5, or cleaved or total cytokeratin 18 may be used to
predict response or sensitivity to a method of treatment described
herein and to identify subjects likely to be responsive to a method
of treatment described herein, such as treatment with a compound of
formula (10), a pharmaceutically acceptable salt thereof, or an
analog thereof. In addition, measuring expression,
post-translational modifications, or activity levels of or
mutations in eIF2-.alpha., ATF4, CHOP, DR5, or cleaved or total
cytokeratin 18 can be used to assess the effectiveness of or
monitor a method of treatment described herein. Furthermore,
measuring expression, post-translational modifications, or activity
levels of or mutations in eIF2-.alpha., ATF4, CHOP, DR5, or cleaved
or total cytokeratin 18 can be used to screen in vivo, in vitro, or
in silico for structurally unrelated anti-cancer molecules. For
example, competition and other assays known in the art may be used
to identify drugs able to outcompete the target interaction with a
higher affinity to compare changes in those levels to the
respective changes produced by a compound of formula (10) or an
analog thereof. Assays can also be performed on living mammalian
cells, which more closely approximate the effects of a particular
serum level of drug in the body, or on microsomal extracts prepared
from cultured cell lines.
[0148] In one embodiment, the subject has, or is at risk of having,
cancer. In one embodiment, the treatment regimen comprises
administering an effective amount of an imipridone, such as ONC201,
or an analog thereof. In one embodiment, the treatment regimen
comprises administering an effective amount of ONC201. In one
embodiment, the treatment regimen comprises administering an
effective amount of a compound of formula (10). In one embodiment,
the compound of formula (10) is a compound of formula (40), e.g., a
compound of formula (45). In one embodiment, a compound of formula
(10) is a compound of formula (50), e.g., a compound formula (55).
In one embodiment, the compound of formula (10) is a compound of
formula (80). In one embodiment, the compound of formula (10) is a
compound of formula (90). In one embodiment, the compound of
formula (10) is a compound of formula (60). In one embodiment,
analogs of compound (1) have a structure selected from the
structures of formula (25), formula (26), formula (27), formula
(28), formula (29), formula (30), or formula (31).
[0149] Levels for a pre-determined standard can be, e.g., the
average or median levels measured in samples from subjects. The
levels for a pre-determined standard can be measured under the same
or substantially similar experimental conditions as in measuring a
sample from a subject. The levels for the pre-determined standard
may be obtained from subjects who are responsive to treatment with
an imipridone, such as ONC201, or an analog thereof. In one
embodiment, the pre-determined standard is obtained from subjects
who are responsive to treatment with the compound, and if the
levels in a sample from a subject are similar to those in the
standard, then the subject can be classified as likely to be
responsive to treatment. The levels for the pre-determined standard
may be obtained from subjects who are not responsive to treatment
with the compound. In one embodiment, the pre-determined standard
is obtained from subjects who are not responsive to treatment with
the compound, and if the levels in a sample from a subject are
different (e.g., up- or down-regulated) than in the pre-determined
standard, then the subject can be classified as likely to be
responsive to treatment. The levels for the pre-determined standard
may be obtained from normal healthy subjects.
[0150] Immunoassays can be used to assay protein or methylation
levels in a sample, including enzyme-linked immunosorbent assay
(ELISA), enzyme-linked immunofiltration assay (ELIFA), flow
cytometry, immunoblot, immunoprecipitation, immunohistochemistry,
immunocytochemistry, luminescent immunoassay (LIA), fluorescent
immunoassay (FIA), and radioimmunoassay. m.sup.6A mRNA methylation
levels can be obtained by methylated RNA immunoprecipitation
(Me-RIP)) or other quantitative biochemical assays known in the
art.
[0151] Nucleic acid mutations can be determined by any of a number
of known procedures. For example, a biologic sample from an
individual can first be obtained. Such biological samples include,
but are not limited to, a bodily fluid (such as urine, saliva,
plasma, or serum) or a tissue sample (such as a buccal tissue
sample or buccal cell). The biologic sample can then 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.
Furthermore, whole or partial genome sequence information can be
used. Such sequences can be determined using standard sequencing
methods including chain-termination (Sanger dideoxynucleotide),
dye-terminator sequencing, and SOLID.TM. sequencing (Applied
Biosystems). Whole genome sequences can be cut by restriction
enzymes or sheared (mechanically) into shorter fragments for
sequencing. DNA sequences can also be amplified using known methods
such as PCR and vector-based cloning methods (e.g., Escherichia
coli). In one embodiment, at least a portion of a subject's genetic
material (e.g., DNA, RNA, mRNA, cDNA, other nucleotide bases or
derivatives of these) is scanned or sequenced using, e.g.,
conventional DNA sequencers or chip-based technologies, to identify
the presence or absence of mutations or copy number variations.
[0152] In one aspect, provided herein are methods of identifying
and treating a subject having a condition and who is likely to be
responsive to a treatment regimen described herein. In one
embodiment, the method comprises (i) identifying whether a subject
having a condition is likely to be responsive to a treatment
regimen described herein; and (ii) treating with the treatment
regimen a subject determined likely to be responsive to that
treatment regimen. In one embodiment, the subject has, or is at
risk of having, cancer. In one embodiment, the treatment regimen
comprises administering an effective amount an imipridone, e.g.,
ONC201 or an analog thereof. In one embodiment, the treatment
regimen comprises administering an effective amount of compound
(1). In one embodiment, the treatment regimen comprises
administering an effective amount of a compound of formula (10). In
one embodiment, the compound of formula (10) is a compound of
formula (40), e.g., a compound of formula (45). In one embodiment,
a compound of formula (10) is a compound of formula (50), e.g., a
compound formula (55). In one embodiment, the compound of formula
(10) is a compound of formula (80). In one embodiment, the compound
of formula (10) is a compound of formula (90). In one embodiment,
the compound of formula (10) is a compound of formula (60). In one
embodiment, analogs of compound (1) have a structure selected from
the structures of formula (25), formula (26), formula (27), formula
(28), formula (29), formula (30), or formula (31).
[0153] Levels for a pre-determined standard can be, e.g., the
average or median levels measured in samples from subjects. The
levels for a pre-determined standard can be measured under the same
or substantially similar experimental conditions as in measuring a
sample from a subject. The levels for the pre-determined standard
may be obtained from subjects who are responsive to treatment with
an imipridone, such as ONC201 or an analog thereof. In one
embodiment, the pre-determined standard is obtained from subjects
who are responsive to treatment with the compound, and if the
levels in a sample from a subject are similar to those in the
standard, then the subject can be classified as likely to be
responsive to treatment. The levels for the pre-determined standard
may be obtained from subjects who are not responsive to treatment
with the compound. In one embodiment, the pre-determined standard
is obtained from subjects who are not responsive to treatment with
the compound, and if the levels in a sample from a subject are
different (e.g., up- or down-regulated) than those in the
pre-determined standard, then the subject can be classified as
likely to be responsive to treatment. The levels for the
pre-determined standard may be obtained from normal healthy
subjects. Immunoassays can be used to assay protein levels in a
sample.
[0154] In one aspect, provided herein are methods of treating and
assessing the effectiveness of a treatment in a subject having a
condition. In one embodiment, the method comprises (i) treating the
subject according to a method of treatment described herein (ii)
assessing as described herein the effectiveness of the treatment.
In one embodiment, the subject has, or is at risk of having,
cancer. In one embodiment, the treatment regimen comprises
administering an effective amount of an imipridone, such as ONC201
or an analog thereof. In one embodiment, the treatment regimen
comprises administering an effective amount of compound (1). In one
embodiment, the treatment regimen comprises administering an
effective amount of a compound of formula (10). In one embodiment,
the compound of formula (10) is a compound of formula (40), e.g., a
compound of formula (45). In one embodiment, a compound of formula
(10) is a compound of formula (50), e.g., a compound formula (55).
In one embodiment, the compound of formula (10) is a compound of
formula (80). In one embodiment, the compound of formula (10) is a
compound of formula (90). In one embodiment, the compound of
formula (10) is a compound of formula (60). In one embodiment,
analogs of compound (1) have a structure selected from the
structures of formula (25), formula (26), formula (27), formula
(28), formula (29), formula (30), or formula (31).
[0155] Other conditions that may be suitable for the methods
described herein include 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, pulmonary heart disease, cardiac
dysrhythmias, inflammatory heart disease, endocarditis,
inflammatory cardiomegaly, myocarditis, valvular heart disease,
cerebrovascular disease, peripheral arterial disease, congenital
heart disease, rheumatic heart disease; Diabetes; and light chain
amyloidosis.
V. COMPOSITIONS
[0156] In one aspect, pharmaceutical compositions are provided,
comprising compounds of formula (10):
##STR00015##
or of formula (1):
##STR00016##
and their pharmaceutically acceptable salts In one embodiment, the
salt is a pharmaceutically acceptable mono-salt of the compound. In
one embodiment, the salt is a pharmaceutically acceptable di-salt
of the compound. In one embodiment, the salt is a pharmaceutically
acceptable mono- or multi-salt (e.g., a di-salt or tri-salt)
thereof selected from the group consisting of hydrochloride,
hydrobromide, hydrogensulphate, sulfates, phosphates, fumarates,
succinates, oxalates and lactates, bisulfates, hydroxyl, tartrate,
nitrate, citrate, bitartrate, carbonate, malate, maleate, fumarate
sulfonate, methylsulfonate, formate, acetate, and carboxylate. In
one embodiment, the salt is a salt selected from the group
consisting of p-toluene-sulfonate, benzenesulfonate, citrate,
methanesulfonate, oxalate, succinate, tartrate, fumarate and
maleate. In one embodiment, the salt is a salt selected from the
group consisting of ammonium, sodium, potassium, calcium,
magnesium, zinc, lithium, and/or with counter-ions such as
methylamino, dimethylamino, diethylamino and triethylamino
counter-ions. In one embodiment, the salt is a. di-hydrochloride
salt or a di-hydrobromide salt.
[0157] Compound (1) (ONC201) has the same chemical structure that
would be revealed by structural analysis (e.g., NMR, X-ray
diffraction) of compound NSC350625, available from the National
Cancer Institute's Developmental Therapeutics Program
Repository.
[0158] In one embodiment, the pharmaceutical composition includes a
di-salt (e.g., a di-hydrochloride salt) of ONC201 or an analog
thereof (e.g., an imipridone). Salts (e.g., di-salts or tri-salts)
of an ONC201 analog can be prepared from an ONC201 analog, which
can be synthesized as described herein, or using standard chemical
synthetic methodology known to one of ordinary skill in the
art.
[0159] In one embodiment, the pharmaceutical composition includes
at least one pharmaceutically acceptable carrier. Suitable
pharmaceutically acceptable carriers, include, but are not limited
to, those in Handbook of Pharmaceutical Excipients, 7th ed., edited
by Raymond C. Rowe et al., American Pharmaceutical Association,
Washington, USA and Pharmaceutical Press, London; and earlier
editions. Exemplary pharmaceutically acceptable carriers, methods
for making pharmaceutical compositions and various dosage forms, as
well as administration modes are well-known in the art, for example
as detailed in Pharmaceutical Dosage Forms: Tablets, edited by
Larry L. Augsburger & Stephen W. Hoag., London: Informa
Healthcare, 2008; and in L. V. Allen, Jr. et al., Ansel's
Pharmaceutical Dosage Forms and Drug Delivery Systems, 8.sup.th
ed., Philadelphia, Pa.: Lippincott, Williams & Wilkins, 2004;
A. R. Gennaro, Remington: The Science and Practice of Pharmacy,
Lippincott Williams & Wilkins, 21.sup.st ed., 2005,
particularly chapter 89; and J. G. Hardman et al., Goodman &
Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill
Professional, 10.sup.th ed., 2001.
[0160] In one embodiment, pharmaceutical compositions are
formulated for ocular administration. In one embodiment,
pharmaceutical compositions are formulated for topical
administration. In one embodiment, pharmaceutical compositions are
formulated as drops, ointments, or liquids. In one embodiment,
pharmaceutical compositions include conventional pharmaceutical
carriers such as aqueous, powdery or oily bases, thickeners.
[0161] In one embodiment, a pharmaceutical composition is a
formulation for intravenous administration. In one embodiment, the
intravenous formulation comprises a compound of formula (10) or a
pharmaceutically acceptable salt thereof dissolved in a solvent. In
one embodiment, the solvent comprises water. In one embodiment, the
intravenous formulation includes the compound or its salt in a
concentration of about 0.05, about 0.25, about 0.5, about 2.5,
about 5, about 25, or about 50 mg/mL. In one embodiment, the
intravenous formulation includes the compound or its salt in a
concentration of from about 0.05, 0.5, or 5 mg/mL to about 1, 10,
or 100 mg/mL. In one embodiment, the intravenous formulation
includes from about 0.005% 0.05%, or 0.5% to about 0.1%, 1%, or 10%
of the compound or its salt. In one embodiment, the intravenous
formulation includes about 0.05%, 0.5%, or 5% of the compound or
its salt. In one embodiment, the intravenous formulation includes a
higher or a lower concentration of the compound or its salt.
[0162] In one embodiment, the intravenous formulation has a pH of
about 3. In one embodiment, the formulation is adjusted to pH 3
with a phosphate buffer. In one embodiment, the intravenous
formulation includes dextrose or sodium chloride. In one
embodiment, the intravenous formulation includes the compound or
its salt in a concentration of about 5 mg/mL and pH 3 and forms a
stable solution. In one embodiment, the intravenous formulation
includes the compound or its salt in a concentration of about 5
mg/mL and pH <5 and forms 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 di-hydrochloride salts
of the compound. In one embodiment, the intravenous formulation
includes the compound or its salt as a 1% solution in a
concentration of about 10 mg/mL. For example, the intravenous
formulation is a solution with a pH of about 3.3. In one
embodiment, the pH is less than 4.0.
[0163] In one embodiment, the pharmaceutical composition further
includes a pharmaceutically acceptable carrier. In one embodiment,
a suitable pharmaceutically acceptable carrier includes an aqueous
carrier. In one embodiment, the aqueous carrier includes sterile
water. In one embodiment, the formulation includes dextrose and/or
sodium. In one embodiment, the pharmaceutically acceptable carrier
includes an oil.
[0164] In one embodiment, an intravenous formulation comprises
ONC201 or an analog thereof or a di-hydrochloride salt thereof
dissolved in water at 25 mg/mL. In one embodiment, the formulation
is adjusted to pH 3 with phosphate buffer. In one embodiment, the
formulation includes dextrose, sodium chloride or both. In one
embodiment, the formulation includes a higher or a lower
concentration of the di-hydrochloride salt of ONC201 or an analog
thereof. In one embodiment, the formulation includes ONC201 or an
analog thereof or a di-hydrochloride salt thereof in a
concentration of about 5 mg/mL. In one embodiment, the formulation
of about 5 mg/mL forms a stable solution and pH 3. In one
embodiment, the formulation of about 5 mg/mL has a pH <5 and
forms a stable solution. In one embodiment, the intravenous
formulation includes ONC201 or an analog thereof or a
di-hydrochloride salt thereof and one or more antioxidants. In one
embodiment, the intravenous formulation includes a mixture of mono-
and di-hydrochloride salts of ONC201 or an analog thereof. In one
embodiment, the intravenous formulation includes ONC201 or an
analog thereof or a di-hydrochloride salt thereof as a 1% solution
in a concentration of about 10 mg/mL. For example, the intravenous
formulation is a solution having a pH of about 3.3. In one
embodiment, the pH is less than 4.0.
[0165] In one embodiment, the intravenous formulation includes from
about 0.5% to about 10% (or from about 5 mg/mL to about 100 mg/mL)
of ONC201 or an analog thereof or a di-salt thereof. In one
embodiment, the formulation includes from about 5% (or about 50
mg/mL) of ONC201 or an analog thereof or a di-salt thereof. In one
embodiment, the intravenous infusion rate may be slowed to decrease
side effects of ONC201 or an analog thereof or a di-salt
thereof.
[0166] In one embodiment, the pharmaceutical composition comprises
about 0.1-99% of an ONC201 salt or an analog thereof; and a
pharmaceutically acceptable carrier, e.g., an oil or sterile water
or other aqueous carrier. In one embodiment, the composition
comprises a mono or di-salt of ONC201 or an analog thereof in a
range of from about 5% to about 50% for oral dosage forms.
[0167] In one embodiment, a pharmaceutical composition includes an
antioxidant. Suitable antioxidants include: ascorbic acid
derivatives such as ascorbic acid, erythorbic acid, sodium
ascorbate, thiol derivatives such as thioglycerol, cysteine,
acetylcysteine, cystine, dithioerythreitol, dithiothreitol,
glutathione, tocopherols, butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), sulfurous acid salts such as sodium sulfate,
sodium bisulfite, acetone sodium bisulfite, sodium metabisulfite,
sodium sulfite, sodium formaldehyde sulfoxylate, and sodium
thiosulfate, nordihydroguaiaretic acid. It should be noted that
antioxidants used for aqueous formulations typically include:
sodium sulphite, sodium metabisulphite, sodium formaldehyde
sulphoxylate and ascorbic acid and combinations thereof, whereas
antioxidants used in oil-based solutions, organic solvents, include
butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA) and
propyl gallate and combinations thereof. In yet other embodiments,
an antioxidant can be one or more of a flavanoid, an isoflavone,
monothioglycerol, L-cysteine, thioglycolic acid,
.alpha.-tocopherol, ascorbic acid 6-palmitate, dihydrolipoic acid,
butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA),
vitamin E, propyl gallate, .beta.-carotene, ascorbic acid.
Antioxidants can typically be used in about 0.1% to 1.0% by weight,
more typically about 0.2%.
[0168] In one embodiment, the pharmaceutical composition includes
an imipridone, such as ONC201 or an analog thereof, or a
pharmaceutically acceptable salt thereof and at least one other
therapeutic agent. For example, the other therapeutic agent is
selected from the group consisting of hormone analogs and
antihormones, aromatase inhibitors, LHRH agonists and antagonists,
inhibitors of growth factors, growth factor antibodies, growth
factor receptor antibodies, tyrosine kinase inhibitors;
antimetabolites; antitumour antibiotics; platinum derivatives;
alkylation agents; antimitotic agents; 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), antiangiogenics (e.g., axitinib, aflibercept, sorafenib,
and regorafenib), amifostin, anagrelid, clodronat, filgrastin,
interferon, interferon .alpha., leucovorin, rituximab,
procarbazine, levamisole, mesna, mitotane, pamidronate and
porfimer, 2-chlorodesoxyadenosine, 2-fluorodesoxy-cytidine,
2-methoxyoestradiol, 2C4,3-alethine, 131-1-TM-601, 3CPA,
7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A 105972, A
204197, abiraterone, aldesleukin, alitretinoin, allovectin-7,
altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037,
AP-5280, apaziquone, apomine, aranose, arglabin, arzoxifene,
atamestane, atrasentan, auristatin PE, ABT-199 (Venetoclax),
ABT-263 (Navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab),
ARRY 162, ARRY 438162, ARRY-300, ARRY-142886/AZD-6244
(selumetinib), ARRY-704/AZD-8330, AR-12, AR-42, AS-703988,
AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026
(primasertib), avastin, AZD-2014, azacytidine, azaepothilone B,
azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576,
bevacizumab, BEZ-235, biricodar dicitrate, BCX-1777, BKM-120,
bleocin, 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, bleomycinic acid, bleomycin
A, bleomycin B, brivanib, bryostatin-1, bortezomib, brostallicin,
busulphan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol,
canertinib, canfosfamide, capecitabine, carboxyphthalatoplatin,
CC1-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime,
ceflatonin, ceftriaxone, celecoxib, celmoleukin, cemadotin,
CH4987655/RO-4987655, chlorotrianisene, cilengitide, ciclosporin,
CDA-II, CDC-394, CKD-602, CKI-27, clofarabin, colchicin,
combretastatin A4, COT inhibitors, CHS-828, CH-5132799, CLL-Thera,
CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibodies,
CP-461, CV-247, cyanomorpholinodoxorubicin, cytarabine, D 24851,
decitabine, deoxorubicin, deoxyrubicin, deoxycoformycin,
depsipeptide, desoxyepothilone B, dexamethasone, dexrazoxanet,
diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10,
doranidazole, DS-7423, E7010, E-6201, edatrexat, edotreotide,
efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509,
enzastaurin, enzalutamide, elsamitrucin, epothilone B, epratuzumab,
ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine,
ethynyloestradiol, exatecan, exatecan mesylate, exemestane,
exisulind, fenretinide, figitumumab, floxuridine, folic acid,
FOLFOX, FOLFOX4, FOLFIRI, formestane, fotemustine, galarubicin,
gallium maltolate, gefinitib, gemtuzumab, gimatecan, glufosfamide,
GCS-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032,
GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100,
gp100-peptide vaccines, GSK-5126766, GSK-690693, GSK-1120212
(trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A,
GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron,
herceptine, hexamethylmelamine, histamine, homoharringtonine,
hyaluronic acid, hydroxyurea, hydroxyprogesterone caproate,
ibandronate, ibrutinib, ibritumomab, idatrexate, idenestrol,
IDN-5109, IGF-1R inhibitors, IMC-1C11, IMC-A12 (cixutumumab),
immunol, indisulam, interferon .alpha.-2a, interferon .alpha.-2b,
pegylated interferon .alpha.-2b, interleukin-2, INK-1117, INK-128,
INSM-18, ionafarnib, ipilimumab, iproplatin, irofulven,
isohomohalichondrin-B, isoflavone, isotretinoin, ixabepilone,
JRX-2, JSF-154, J-107088, conjugated oestrogens, kahalid F,
ketoconazole, KW-2170, KW-2450, lobaplatin, leflunomide,
lenograstim, leuprolide, leuporelin, lexidronam, LGD-1550,
linezolid, lutetium texaphyrin, lometrexol, losoxantrone, LU
223651, lurtotecan, LY-S6AKT1, LY-2780301, mafosfamide, marimastat,
mechloroethamine, MEK inhibitors, MEK-162, methyltestosteron,
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, norelin,
N-acetylcysteine, 06-benzylguanine, oblimersen, omeprazole,
oncophage, oncoVEXGM-CSF, ormiplatin, ortataxel, OX44 antibodies,
OSI-027, OSI-906 (linsitinib), 4-1BB antibodies, oxantrazole,
oestrogen, panitumumab, patupilone, pegfilgrastim, PCK-3145,
pegfilgrastim, PBI-1402, PBI-05204, PD0325901, PD-1 antibodies,
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 inhibitors, PI3K/mTOR inhibitors, PG-TXL, PG2,
PLX-4032/RO-5185426 (vemurafenib), PLX-3603/RO-5212054, PT-100,
PWT-33597, PX-866, picoplatin, pivaloyloxymethylbutyrate,
pixantrone, phenoxodiol O, PKI166, plevitrexed, plicamycin,
polyprenic acid, porfiromycin, prednisone, prednisolone, quinamed,
quinupristin, R115777, RAF-265, ramosetron, ranpirnase,
RDEA-119/BAY 869766, RDEA-436, rebeccamycin analogs, receptor
tyrosine kinase (RTK) inhibitors, regorafenib, revimid, RG-7167,
RG-7304, RG-7421, RG-7321, RG 7440, rhizoxin, rhu-MAb, rinfabate,
risedronate, rituximab, robatumumab, rofecoxib, RO-31-7453,
RO-5126766, RO-5068760, RPR 109881A, rubidazone, rubitecan,
R-flurbiprofen, RX-0201, S-9788, sabarubicin, SAHA, sargramostim,
satraplatin, SB 408075, Se-015/Ve-015, SU5416, SU6668, SDX-101,
semustin, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897,
SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine,
suberanilohydroxamic acid, sutent, T 900607, T 138067, TAK-733,
TAS-103, tacedinaline, talaporf in, Tarceva, tariquitar, tasisulam,
taxotere, taxoprexin, tazarotene, tegafur, temozolamide,
tesmilifene, testosterone, testosterone propionate, tesmilifene,
tetraplatin, tetrodotoxin, tezacitabine, thalidomide, theralux,
therarubicin, thymalfasin, thymectacin, tiazofurin, tipifarnib,
tirapazamine, tocladesine, tomudex, toremofin, trabectedin,
TransMID-107, transretinic acid, traszutumab, tremelimumab,
tretinoin, triacetyluridine, triapine, triciribine, trimetrexate,
TLK-286TXD 258, tykerb/tyverb, urocidin, valrubicin, vatalanib,
vincristine, vinflunine, 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.
[0169] In one embodiment, the other therapeutic agent comprises a
hormone analog, an antihormone or both selected from tamoxifen,
toremifene, raloxifene, fulvestrant, megestrol acetate, flutamide,
nilutamide, bicalutamide, aminoglutethimide, cyproterone acetate,
finasteride, buserelin acetate, fludrocortisone, fluoxymesterone,
medroxy-progesterone, octreotide, and combinations thereof. In one
embodiment, the other therapeutic agent comprises one or more LHRH
agonists and/or antagonists selected from goserelin acetate,
luprolide acetate, triptorelin pamoate and combinations thereof and
wherein the LHRH antagonists are selected from Degarelix,
Cetrorelix, Abarelix, Ozarelix, Degarelix combinations thereof. In
one embodiment, the other therapeutic agent comprises one or more
growth factor inhibitors selected from inhibitors of: platelet
derived growth factor (PDGF), fibroblast growth factor (FGF),
vascular endothelial growth factor (VEGF), epidermal growth factor
(EGF), insuline-like growth factors (IGF), human epidermal growth
factor (HER) and hepatocyte growth factor (HGF). In one embodiment,
the other therapeutic agent comprises one or more inhibitors of the
human epidermal growth factor selected from HER2, HER3, and HER4.
In one embodiment, the other therapeutic agent comprises one or
more tyrosine kinase inhibitors selected from cetuximab, gefitinib,
imatinib, lapatinib and trastuzumab, and combinations thereof. In
one embodiment, the other therapeutic agent comprises one or more
aromatase inhibitors selected from anastrozole, letrozole,
liarozole, vorozole, exemestane, atamestane, and combinations
thereof. In one embodiment, the other therapeutic agent comprises
one or more antimetabolites which are antifolates selected from
methotrexate, raltitrexed, and pyrimidine analogs. In one
embodiment, the other therapeutic agent comprises one or more
antimetabolites which are pyrimidine analogs selected from
5-fluorouracil, capecitabin and gemcitabin. In one embodiment, the
other therapeutic agent comprises 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
agent comprises one or more antitumour antibiotics selected from
anthracyclins, doxorubicin, daunorubicin, epirubicin and
idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin,
streptozocin and combinations thereof. In one embodiment, the other
therapeutic agent comprises one or more platinum derivatives
selected from cisplatin, oxaliplatin, carboplatin and combinations
thereof. In one embodiment, the other therapeutic agent comprises
one or more alkylation agents selected from estramustin,
meclorethamine, melphalan, chlorambucil, busulphan, dacarbazin,
cyclophosphamide, ifosfamide, temozolomide, nitrosoureas, and
combinations thereof. In one embodiment, the other therapeutic
agent comprises nitrosoureas selected from carmustin, lomustin,
thiotepa, and combinations thereof. In one embodiment, the other
therapeutic agent comprises antimitotic agents selected from Vinca
alkaloids and taxanes. In one embodiment, the other therapeutic
agent comprises one or more taxanes selected from paclitaxel,
docetaxel, and combinations thereof. In one embodiment, the other
therapeutic agent comprises one or more Vinca alkaloids selected
from vinblastine, vindesin, vinorelbin, vincristine, and
combinations thereof. In one embodiment, the other therapeutic
agent comprises one or more topoisomerase inhibitors which are
epipodophyllotoxins. In one embodiment, the other therapeutic agent
comprises one or more epipodophyllotoxins selected from etoposide
and etopophos, teniposide, amsacrin, topotecan, irinotecan,
mitoxantron, and combinations thereof. In one embodiment, the other
therapeutic agent comprises one or more serine/threonine kinase
inhibitors 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,
inhibitors of CDKs, Aurora kinase inhibitors, and combinations
thereof. In one embodiment, the other therapeutic agent comprises
one or more tyrosine kinase inhibitors which are PTK2/FAK
inhibitors. In one embodiment, the other therapeutic agent
comprises one or more protein protein interaction inhibitors
selected from IAP, Mcl-1, MDM2/MDMX and combinations thereof. In
one embodiment, the other therapeutic agent comprises one or more
rapamycin analogs selected from everolimus, temsirolimus,
ridaforolimus, sirolimus, and combinations thereof. In one
embodiment, the other therapeutic agent comprises one or more
therapeutic agents selected from amifostin, anagrelid, clodronat,
filgrastin, interferon, interferon .alpha., leucovorin, rituximab,
procarbazine, levamisole, mesna, mitotane, pamidronate and
porfimer, and combinations thereof. In one embodiment, the other
therapeutic agent comprises one or more therapeutic agents selected
from 2-chlorodesoxyadenosine, 2-fluorodesoxy-cytidine,
2-methoxyoestradiol, 2C4,3-alethine, 131-1-TM-601, 3CPA,
7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A 105972, A
204197, abiraterone, aldesleukin, alitretinoin, allovectin-7,
altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037,
AP-5280, apaziquone, apomine, aranose, arglabin, arzoxifene,
atamestane, atrasentan, auristatin PE, ABT-199 (Venetoclax),
ABT-263 (Navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab),
ARRY 162, ARRY 438162, ARRY-300, ARRY-142886/AZD-6244
(selumetinib), ARRY-704/AZD-8330, AR-12, AR-42, AS-703988,
AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026
(primasertib), avastin, AZD-2014, azacytidine, azaepothilone B,
azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576,
bevacizumab, BEZ-235, biricodar dicitrate, BCX-1777, BKM-120,
bleocin, 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, bleomycinic acid, bleomycin
A, bleomycin B, brivanib, bryostatin-1, bortezomib, brostallicin,
busulphan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol,
canertinib, canfosfamide, capecitabine, carboxyphthalatoplatin,
CC1-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime,
ceflatonin, ceftriaxone, celecoxib, celmoleukin, cemadotin,
CH.sub.4987655/RO-4987655, chlorotrianisene, cilengitide,
ciclosporin, CDA-II, CDC-394, CKD-602, CKI-27, clofarabin,
colchicin, combretastatin A4, COT inhibitors, CHS-828, CH-5132799,
CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal
antibodies, CP-461, CV-247, cyanomorpholinodoxorubicin, cytarabine,
D 24851, decitabine, deoxorubicin, deoxyrubicin, deoxycoformycin,
depsipeptide, desoxyepothilone B, dexamethasone, dexrazoxanet,
diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10,
doranidazole, DS-7423, E7010, E-6201, edatrexat, edotreotide,
efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509,
enzastaurin, enzalutamide, elsamitrucin, epothilone B, epratuzumab,
ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine,
ethynyloestradiol, exatecan, exatecan mesylate, exemestane,
exisulind, fenretinide, figitumumab, floxuridine, folic acid,
FOLFOX, FOLFOX4, FOLFIRI, formestane, fotemustine, galarubicin,
gallium maltolate, gefinitib, gemtuzumab, gimatecan, glufosfamide,
GCS-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032,
GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100,
gp100-peptide vaccines, GSK-5126766, GSK-690693, GSK-1120212
(trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A,
GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron,
herceptine, hexamethylmelamine, histamine, homoharringtonine,
hyaluronic acid, hydroxyurea, hydroxyprogesterone caproate,
ibandronate, ibrutinib, ibritumomab, idatrexate, idenestrol,
IDN-5109, IGF-1R inhibitors, IMC-1C11, IMC-A12 (cixutumumab),
immunol, indisulam, interferon .alpha.-2a, interferon .alpha.-2b,
pegylated interferon .alpha.-2b, interleukin-2, INK-1117, INK-128,
INSM-18, ionafarnib, ipilimumab, iproplatin, irofulven,
isohomohalichondrin-B, isoflavone, isotretinoin, ixabepilone,
JRX-2, JSF-154, J-107088, conjugated oestrogens, kahalid F,
ketoconazole, KW-2170, KW-2450, lobaplatin, leflunomide,
lenograstim, leuprolide, leuporelin, lexidronam, LGD-1550,
linezolid, lutetium texaphyrin, lometrexol, losoxantrone, LU
223651, lurtotecan, LY-S6AKT1, LY-2780301, mafosfamide, marimastat,
mechloroethamine, MEK inhibitors, MEK-162, methyltestosteron,
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, norelin,
N-acetylcysteine, 06-benzylguanine, oblimersen, omeprazole,
oncophage, oncoVEXGM-CSF, ormiplatin, ortataxel, OX44 antibodies,
OSI-027, OSI-906 (linsitinib), 4-1BB antibodies, oxantrazole,
oestrogen, panitumumab, patupilone, pegfilgrastim, PCK-3145,
pegfilgrastim, PBI-1402, PBI-05204, PD0325901, PD-1 antibodies,
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 inhibitors, PI3K/mTOR inhibitors, PG-TXL, PG2,
PLX-4032/RO-5185426 (vemurafenib), PLX-3603/RO-5212054, PT-100,
PWT-33597, PX-866, picoplatin, pivaloyloxymethylbutyrate,
pixantrone, phenoxodiol O, PKI166, plevitrexed, plicamycin,
polyprenic acid, porfiromycin, prednisone, prednisolone, quinamed,
quinupristin, R115777, RAF-265, ramosetron, ranpirnase,
RDEA-119/BAY 869766, RDEA-436, rebeccamycin analogs, receptor
tyrosine kinase (RTK) inhibitors, revimid, RG-7167, RG-7304,
RG-7421, RG-7321, RG 7440, rhizoxin, rhu-MAb, rinfabate,
risedronate, rituximab, robatumumab, rofecoxib, RO-31-7453,
RO-5126766, RO-5068760, RPR 109881A, rubidazone, rubitecan,
R-flurbiprofen, RX-0201, S-9788, sabarubicin, SAHA, sargramostim,
satraplatin, SB 408075, Se-015/Ve-015, SU5416, SU6668, SDX-101,
semustin, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897,
SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine,
suberanilohydroxamic acid, sutent, T 900607, T 138067, TAK-733,
TAS-103, tacedinaline, talaporf in, Tarceva, tariquitar, tasisulam,
taxotere, taxoprexin, tazarotene, tegafur, temozolamide,
tesmilifene, testosterone, testosterone propionate, tesmilifene,
tetraplatin, tetrodotoxin, tezacitabine, thalidomide, theralux,
therarubicin, thymalfasin, thymectacin, tiazofurin, tipifarnib,
tirapazamine, tocladesine, tomudex, toremofin, trabectedin,
TransMID-107, transretinic acid, traszutumab, tremelimumab,
tretinoin, triacetyluridine, triapine, triciribine, trimetrexate,
TLK-286TXD 258, tykerb/tyverb, urocidin, valrubicin, vatalanib,
vincristine, vinflunine, 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.
[0170] In one embodiment, the other therapeutic agent comprises a
steroid, including dexamethasone, prednisolone, methyl
prednisolone, prednisone, hydrocortisone, triamcinolone,
betamethasone, and cortivazol. In one embodiment, the other
therapeutic agent comprises an anti-emetic, Anti-emetics include,
but are not limited to, 5-HT3 receptor agonists (e.g., dolasetron,
granisetron, ondansetron, tropisetron, palonosetron, and
mirtazapine), dopamine agonists (e.g., domperidone, olanzapine,
droperidol, haloperidol, chlorpromazine, prochlorperazine,
alizapride, prochlorperazine, and metoclopramide), NK1 receptor
antagonists (e.g., aprepitant and casopitant), antihistamines (such
as cyclizine, diphenhydramine, dimenhydrinate, doxylamine,
meclizine, promethazine, hydroxyzine), cannabinoids (e.g.,
cannabis, dronabinol, nabilone, and sativex), benzodiazepines
(e.g., midazolam and lorazepam), anticholinergics (e.g., hyoscine),
trimethobenzamide, ginger, emetrol, propofol, peppermint, muscimol,
and ajwain.
[0171] In one embodiment, the other therapeutic agent comprises an
anti-cancer agent, which includes a mitotic inhibitor. In one
embodiment, the mitotic inhibitor includes a taxane. In one
embodiment, the mitotic inhibitor includes a taxane selected from
paclitaxel and docetaxel.
[0172] In one embodiment, the pharmaceutical composition includes
an imipridone, such as ONC201, or an analog thereof, or a
pharmaceutically acceptable salt thereof; and at least one
anti-cancer agent, which includes one or more of acivicin,
aclarubicin, acodazole, acronine, adozelesin, aldesleukin,
alitretinoin, allopurinol, altretamine, ambomycin, ametantrone,
amifostine, aminoglutethimide, amsacrine, anastrozole, anthramycin,
arsenic trioxide, asparaginase, asperlin, azacitidine, azetepa,
azotomycin, batimastat, benzodepa, bevacizumab, bicalutamide,
bisantrene, bisnafide dimesylate, bizelesin, bleomycin, brequinar,
bropirimine, busulfan, cactinomycin, calusterone, capecitabine,
caracemide, carbetimer, carboplatin, carmustine, carubicin,
carzelesin, cedefingol, celecoxib, chlorambucil, cirolemycin,
cisplatin, cladribine, crisnatol mesylate, cyclophosphamide,
cytarabine, dacarbazine, dactinomycin, daunorubicin, decitabine,
dexormaplatin, dezaguanine, dezaguanine mesylate, diaziquone,
docetaxel, doxorubicin, droloxifene, dromostanolone, duazomycin,
edatrexate, eflomithine, elsamitrucin, enloplatin, enpromate,
epipropidine, epirubicin, erbulozole, esorubicin, estramustine,
etanidazole, etoposide, etoprine, fadrozole, fazarabine,
fenretinide, floxuridine, fludarabine, fluorouracil, flurocitabine,
fosquidone, fostriecin, fulvestrant, gemcitabine, hydroxyurea,
idarubicin, ifosfamide, ilmofosine, interleukin II (IL-2, including
recombinant interleukin II or rIL2), interferon .alpha.-2a,
interferon .alpha.-2b, interferon .alpha.-n1, interferon
.alpha.-n3, interferon .beta.-Ia, interferon gamma-Ib, iproplatin,
irinotecan, lanreotide, letrozole, leuprolide, liarozole,
lometrexol, lomustine, losoxantrone, masoprocol, maytansine,
mechlorethamine hydrochlride, megestrol, melengestrol acetate,
melphalan, menogaril, mercaptopurine, methotrexate, metoprine,
meturedepa, mitindomide, mitocarcin, mitocromin, mitogillin,
mitomalcin, mitomycin, mitosper, mitotane, mitoxantrone,
mycophenolic acid, nelarabine, nocodazole, nogalamycin,
ormnaplatin, oxisuran, paclitaxel, pegaspargase, peliomycin,
pentamustine, peplomycin, perfosfamide, pipobroman, piposulfan,
piroxantrone hydrochloride, plicamycin, plomestane, porfimer,
porfiromycin, prednimustine, procarbazine, puromycin, pyrazofurin,
riboprine, rogletimide, safingol, semustine, simtrazene,
sparfosate, sparsomycin, spirogermanium, spiromustine, spiroplatin,
streptonigrin, streptozocin, sulofenur, talisomycin, tamoxifen,
tecogalan, tegafur, teloxantrone, temoporfin, teniposide,
teroxirone, testolactone, thiamiprine, thioguanine, thiotepa,
tiazofurin, tirapazamine, topotecan, toremifene, trestolone,
triciribine, trimetrexate, triptorelin, tubulozole, uracil mustard,
uredepa, vapreotide, verteporfin, vinblastine, vincristine sulfate,
vindesine, vinepidine, vinglycinate, vinleurosine, vinorelbine,
vinrosidine, vinzolidine, vorozole, zeniplatin, zinostatin,
zoledronate, zorubicin and combinations thereof.
[0173] Examples of suitable anti-cancer agents include those
described Goodman and Gilman's The Pharmacological Basis of
Therapeutics, 12.sup.th Ed., edited by Laurence Brunton, Bruce
Chabner, Bjorn Knollman, McGraw Hill Professional, 2010.
[0174] In some exemplary embodiments, the pharmaceutical
composition includes a salt (e.g., a mono- or di-salt) of an
imipridone, e.g., ONC201, or an analog thereof and at least one
other therapeutic agent, where 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, l6 kDa prolactin
fragment, laminin peptides, fibronectin peptides, tissue
metalloproteinase inhibitors (TIMP 1, 2, 3, 4), plasminogen
activator inhibitors (PAI-1, -2), tumor necrosis factor .alpha.,
(high dose, invitro), TGF-.beta.1, interferons (IFN-.alpha.,
-.beta., .gamma.), ELR-CXC chemokines, IL-12; SDF-1; MIG; platelet
factor 4 (PF-4); IP-10, thrombospondin (TSP), SPARC,
2-methoxyoestradiol, proliferin-related protein, suramin,
sorafenib, regorafenib, thalidomide, cortisone, linomide,
fumagillin (AGM-1470; TNP-470), tamoxifen, retinoids, CM101,
dexamethasone, leukemia inhibitory factor (LIF), hedgehog inhibitor
and combinations thereof.
[0175] A pharmaceutical combination can include first and second
therapeutic agents in any desired proportions provided that the
synergistic or cooperative effect still occurs. A synergistic
pharmaceutical combination preferably contains the first and second
therapeutic agents in a ratio of from about 1:9 to about 9:1. In
one embodiment, a synergistic combination contains the first and
second therapeutic agents in a ratio of from about 1:8 to about
8:1, from 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 combination contains the therapeutic agents in a
ratio of approximately 1:1.
[0176] In one embodiment, the second therapeutic agent is selected
from Allopurinol, Arsenic Trioxide, Azacitidine, Bortezomib,
Bevacizumab, Capecitabine, Carboplatin, Celecoxib, Chlorambucil,
Clofarabine, Cytarabine, Dacarbazine, Daunorubicin HCl, Docetaxel,
Doxorubicin HCl, Floxuridine, Gemcitabine HCl, Hydroxyurea,
Ifosfamide, Imatinib Mesylate, Ixabepilone, Lenalidomide, Megestrol
acetate, Methotrexate, Mitotane, Mitoxantrone HCl, Oxaliplatin,
Paclitaxel, Pralatrexate, Romidepsin, Sorafenib, Streptozocin,
Tamoxifen Citrate, Topotecan HCl, Tretinoin, Vandetanib,
Vismodegib, Vorinostat, and combinations thereof.
[0177] In one embodiment, the second therapeutic agent comprises a
small molecule multi-kinase inhibitor, e.g., sorafenib or
regorafenib. In one embodiment, the second therapeutic agent
comprises a Hedgehog Pathway Inhibitor, e.g., vismodegib. In one
embodiment, the second therapeutic agent includes a drug selected
from Table 2 below.
TABLE-US-00002 TABLE 2 Classes Of Drugs Classes of drugs Examples
Purine analogs allopurinol, oxypurinol, clofarabine, and tisopurine
Pyrimidine 5-fluorouracil, Floxuridine (FUDR), capecitabine,
analogs cytarabine, 6-azauracil (6-AU), and gemcitabine (Gemzar)
Proteasome bortezomib, carfilzomib, cecliranib, disulfiram,
inhibitors epigallocatechin-3-gallate, salinosporamide A, ONCX
0912, CEP-18770, MLN9708, epoxomicin, and MG132. Anti- bevacizumab,
aflibercept, sunitinib, sorafenib, pazopanib, angiogenic
vandetanib, cabozantinib, axitinib, ponatinib, regorafenib,
ranibizumab, lapatinib, and vandetanib. Platinum-based cisplatin,
carboplatin, oxaliplatin, satraplatin, picoplatin, antineo-
nedaplatin, and triplatin. plastic drugs COX-2 celecoxib,
valdecoxib (Bextra), parecoxib (Dynastat), inhibitors lumiracoxib,
etoricoxib, and rofecoxib. Nitrogen cyclophosphamide, chlorambucil,
uramustine, ifosfamide, mustards melphalan, bendamustine, and
mustine. Alkylating cyclophosphamide, mechlorethamine or mustine
(HN2) agents (trade name Mustardgen), uramustine or uracil mustard,
melphalan, chlorambucil, ifosfamide, bendamustine, carmustine,
lomustine, streptozocin, and busulfan. Anthracyclines Daunorubicin
(Daunomycin), Daunorubicin (liposomal), Doxorubicin (Adriamycin),
Doxorubicin (liposomal), Epirubicin, Idarubicin, Valrubicin, and
Mitoxantrone. Taxanes Paclitaxel (Taxol), Docetaxel (Taxotere), and
albumin-bound paclitaxel (Abraxane). Nucleotide methotrexate,
pralatrexate, hydroxyurea, and synthesis 5-fluorodeoxyuridine,
3,4-dihydroxybenzylamine inhibitor Bcr-abl imatinib, nilotinib,
dasatinib, bosutinib and ponatinib. inhibitors Other arsenic
trioxide, thalidomide, revlimid, and mitotane. Topoisomerase
amsacrine, etoposide, etoposide phosphate, teniposide, inhibitor
doxorubicin, Topotecan (Hycamtin), Irinotecan (CPT-11, Camptosar),
Exatecan, Lurtotecan, ST 1481, CKD 602, ICRF-193, and genistein.
HDAC Vorinostat (SAHA), Romidepsin (Istodax), Panobinostat
inhibitors (LBH589), Valproic acid (as Mg valproate), Belinostat
(PXD101), Mocetinostat (MGCD0103), Abexinostat (PCI-24781),
Entinostat (MS-275), SB939, Resminostat (4SC-201), Givinostat,
Quisinostat (JNJ-26481585), CUDC-101, AR-42, CHR-2845, CHR-3996,
4SC-202, CG200745, ACY-1215, ME-344, sulforaphane, Kevetrin, and
ATRA. Multi-kinase sorafenib, regorafenib, and vandetanib.
inhibitors Hormone tamoxifen, toremifene, Arimidex (anastrozole),
Aromasin therapies (exemestane), Femara (letrozole), and
Fulvestrant (Faslodex). Hedgehog vismodegib, BMS-833923, IPI-926,
LDE-225, signaling PF-04449913, LEQ 506, and TAK-441. Inhibitors
Checkpoint Opdivo (nivolumab), Durvalumab (Medi4736), Keytruda
Inhibitors (pembrolizumab, MK3475), BGB-A317, AMP-224, PDR001, REGN
281, Atezolizumab (MPDL3280A), Pidilizumab (BMS-936559, CT-011,
ONO-4538), Avelumab (MSB0010718 C), Yervoy (ipilimumab),
tremelimumab BCL2 AT-101, Bcl-2/xL inhibitor, Navitoclax (ABT-263),
Inhibitors Venetoclax (ABT-199), Apogossypol, PTN1258, obatoclax,
G3139
[0178] In one embodiment, the second therapeutic agent includes
drugs that target tumor necrosis factor-related apoptosis-inducing
ligand (TRAIL) receptors. In one embodiment, the second therapeutic
agent includes a recombinant TRAIL or an agonistic 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 signaling 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 an active agent selected from Camptothecin, 5-FU,
capecitabine, cisplatin, doxorubicin, irinotecan, paclitaxel,
cisplatin, bortezomib, BH3I-2, rituximab, radiation, triterpenoids,
sorafenib, gemcitabine, HDAC inhibitors, carboplatin, T-101 (a
gossypol derivate), ABT-263, ABT-737, and GX-15-070 (obatoclax),
vorinostat, cetuximab, panitumumab, bevacizumab, ganitumab,
interferon gamma, sorafenib, XIAP antagonists, Bcl-2 antagonists,
and Smac mimetics.
VI. DOSE
[0179] In one embodiment, a pharmaceutical composition comprises an
imipridone, such as ONC201, or an analog thereof, or a
pharmaceutically acceptable salt thereof in a dose ranging from
about 40, 50, 60, or 100 mg to about 2000 mg; from about 4, 5, 6,
or 10 mg to about 200 mg; or from about 0.4, 0.5, 0.6, or 1 mg to
about 20 mg where the weight can be based on the compound in its
free base form. In one embodiment, a pharmaceutical composition
comprises an imipridone, such as ONC201, or an analog thereof, or a
pharmaceutically acceptable salt thereof in a dose level ranging
from 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, a pharmaceutical
composition comprises an imipridone, such as ONC201, or an analog
thereof, or a pharmaceutically acceptable salt thereof in a dose
level ranging from 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, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 mg. In one
embodiment, a pharmaceutical composition comprises an imipridone,
such as ONC201, or an analog thereof, or a pharmaceutically
acceptable salt thereof in a dose level ranging from 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; from about 6
mg to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130,
140, 150, 160, 170, 180, 190, or 200 mg; or from about 0.6 mg to
about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,
19, or 20 mg. In one embodiment, a pharmaceutical composition
comprises an imipridone, such as ONC201, or an analog thereof, or a
pharmaceutically acceptable salt thereof in a dose level ranging
from 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; from about 10 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 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, a
pharmaceutical composition comprises an imipridone, such as ONC201,
or an analog thereof, or a pharmaceutically acceptable salt thereof
in a dose level ranging from 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; from 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 from about 2 mg to about 3, 4, 5, 6, 7, 8, 9, 10, 11,
12, 13, 14, 15, 16, 17, 18, 19, or 20 mg, based on the compound in
its free base form. In one embodiment, a pharmaceutical composition
comprises an imipridone, such as ONC201, or an analog thereof, or a
pharmaceutically acceptable salt thereof in a dose level ranging
from about 400 mg to about 500, 600, 700, 800, 900, 1000, 1100,
1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, or 2000 mg; from
about 40 mg to about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140,
150, 160, 170, 180, 190, or 200 mg; or from about 4 mg to about 5,
6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg based
on the compound in its free base form. In one embodiment, a
pharmaceutical composition comprises an imipridone, such as ONC201,
or an analog thereof, or a pharmaceutically acceptable salt thereof
in a dose level ranging from about 50 mg to about 60, 70, 80, 90,
or 100 mg; from about 60 mg to about 70, 80, 90, or 100 mg; from
about 70 mg to about 80, 90 or 100 mg, from about 80 mg to about 90
or 100 mg; from about 90 mg to about 100 mg; from about 5 mg to
about 6, 7, 8, 9, or 10 mg; from about 6 mg to about 7, 8, 9, or 10
mg; from about 7 mg to about 8, 9 or 10 mg, from about 8 mg to
about 9 or 10 mg; from about 9 mg to about 10 mg; from about 0.5 mg
to about 0.6, 0.7, 0.8, 0.9, or 1 mg; from about 0.6 mg to about
0.7, 0.8, 0.9, or 1 mg; from about 0.7 mg to about 0.8, 0.9 or 1
mg, from about 0.8 mg to about 0.9 or 1 mg; or from about 0.9 mg to
about 1 mg.
[0180] In one embodiment, a pharmaceutical composition comprises an
imipridone, such as ONC201, or an analog thereof, or a
pharmaceutically acceptable salt thereof in a dose ranging from
about 1 mg/kg to about 40 mg/kg; 0.1 mg/kg to about 4 mg/kg; or
0.01 mg/kg to about 0.40 mg/kg. In one embodiment, a pharmaceutical
composition comprises an imipridone, such as ONC201, or an analog
thereof, or a pharmaceutically acceptable salt thereof in a dose
level ranging from about 1, 2, 3, 4, 5, 6, 7, 8, or 9 mg/kg to
about 10, 20, 30, or 40 mg/kg; from about 10, 11, 12, 13, 14, 15,
16, 17, 18, or 19 mg/kg to about 20, 30, or 40 mg/kg; from about
20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 mg/kg to about 30 or 40
mg/kg; from 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, 2.6, 2.7, 2.8,
or 2.9 mg/kg to about 3 or 4 mg/kg; or from about 3.0, 3.1, 3.2,
3.3, 3.4, 3.5, 3.6, 3.7, 3.8, or 3.9 mg/kg to about 4 mg/kg; from
about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 mg/kg to
about 0.10, 0.20, 0.30, or 0.40 mg/kg; from about 0.10, 0.11, 0.12,
0.13, 0.14, 0.15, 0.16, 0.17, 0.18, or 0.19 mg/kg to about 0.20,
0.30, or 0.40 mg/kg; from about 0.20, 0.21, 0.22, 0.23, 0.24, 0.25,
0.26, 0.27, 0.28, or 0.29 mg/kg to about 0.30 or 0.40 0.mg/kg; or
from about 0.30, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, or
0.39 mg/kg to about 0.40 mg/kg.
[0181] In one embodiment, a pharmaceutical composition comprises an
imipridone, such as ONC201, or an analog thereof, or a
pharmaceutically acceptable salt thereof in a dose ranging from
about 37.5 mg/m.sup.2 to about 1500 mg/m.sup.2; from about 3.75
mg/m.sup.2 to about 150 mg/m.sup.2; or from about 0.4 mg/m.sup.2 to
about 15 mg/m.sup.2 In one embodiment, a pharmaceutical composition
comprises an imipridone, such as ONC201, or an analog thereof, or a
pharmaceutically acceptable salt thereof in a dose 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, 400, 405, 410, 415, 420, 425, 430,
435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495,
500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560,
565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625,
630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690,
695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755,
760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820,
825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885,
890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950,
955, 960, 965, 970, 975, 980, 985, 990, 995, 1000, 1005, 1010,
1015, 1020, 1025, 1030, 1035, 1040, 1045, 1050, 1055, 1060, 1065,
1070, 1075, 1080, 1085, 1090, 1095, 1100, 1105, 1110, 1115, 1120,
1125, 1130, 1135, 1140, 1145, 1150, 1155, 1160, 1165, 1170, 1175,
1180, 1185, 1190, 1195, 1200, 1205, 1210, 1215, 1220, 1225, 1230,
1235, 1240, 1245, 1250, 1255, 1260, 1265, 1270, 1275, 1280, 1285,
1290, 1295, 1300, 1305, 1310, 1315, 1320, 1325, 1330, 1335, 1340,
1345, 1350, 1355, 1360, 1365, 1370, 1375, 1380, 1385, 1390, 1395,
1400, 1405, 1410, 1415, 1420, 1425, 1430, 1435, 1440, 1445, 1450,
1455, 1460, 1465, 1470, 1475, 1480, 1485, 1490, 1495 mg/m.sup.2 to
about 1500 mg/m.sup.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.sup.2 to about 150 mg/m.sup.2; or from about 0.5, 1, 1.5, 2,
2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10,
10.5, 11, 111, 11.5, 12, 12.5, 13, 13.5, 14, or 14.5 mg/m.sup.2 to
about 15 mg/m.sup.2.
VII. DOSAGE FORMS
[0182] Suitable pharmaceutical compositions for use in the methods
described herein can be formulated into a dosage form that can be
administered to a patient. In one embodiment, a pharmaceutical
composition is in the form of an oral dosage unit or parenteral
dosage unit. In one embodiment, a pharmaceutical composition is in
the form of an oral dosage unit. In one embodiment, an oral dosage
unit is fractionated into several, smaller doses, which are
administered to a subject over a predetermined period of time in
order to reduce toxicity of a therapeutic agent being administered.
In one embodiment, an oral dosage unit is administered by a tablet
or capsule comprising a controlled release formulation that can
include a plurality of particles, granules, pellets, minitablets or
tablets. In one embodiment, the pharmaceutical composition is in
the form of a parenteral dosage unit. In one embodiment, the
parenteral dosage unit is selected from the group consisting of
intravenous (IV), subcutaneous (SC), and intramuscular (M), rectal
(PR) and transdermal dosage units. In one embodiment, the
composition is in a dosage form selected from the group consisting
of sterile solutions, suspensions, suppositories, tablets and
capsules. In one embodiment, the composition is an oral dosage form
selected from the group consisting of a tablet, caplet, capsule,
lozenge, syrup, liquid, suspension and elixir. 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
semi-solids.
[0183] In one embodiment, suitable forms of pharmaceutical
compositions for use in the methods described herein include
dermatological compositions adapted for cutaneous topical
administration. For example, dermatological compositions include a
cosmetically or pharmaceutically acceptable medium. Dermatological
compositions for topical administration can include ointments,
lotions, creams, gels, drops, suppositories, sprays, liquids and
powders. In one embodiment, conventional pharmaceutical carriers,
aqueous, powder or oily bases, thickeners, skin enhancers can be
necessary or desirable and therefore used. Examples of suitable
enhancers include ethers such as diethylene glycol monoethyl ether
(available commercially as TRANSCUTOL.RTM.) and diethylene glycol
monomethyl ether; surfactants such as sodium laurate, sodium lauryl
sulfate, cetyltrimethylammonium bromide, benzalkonium chloride,
Poloxamer (231, 182, 184), Tween (20, 40, 60, 80), and lecithin
(U.S. Pat. No. 4,783,450); alcohols such as ethanol, propanol,
octanol, benzyl alcohol; polyethylene glycol and esters thereof
such as polyethylene glycol monolaurate; amides and other
nitrogenous compounds such as urea, dimethylacetamide (DMA),
dimethylformamide (DMF), 2-pyrrolidone, 1-methyl-2-pyrrolidone,
ethanolamine, diethanolamine and triethanolamine; terpenes;
alkanones; and organic acids, particularly citric acid and succinic
acid. AZONE.RTM. and sulfoxides such as DMSO and C0MSO may also be
used, but are less preferred.
[0184] In one embodiment, the pharmaceutical composition is in a
dosage form selected from the group consisting of sustained release
forms, controlled release forms, delayed release forms and response
release forms.
VIII. METHODS OF USE
[0185] The compositions and methods described herein have utility
in treating many disease conditions, 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, desmoplastic round cell tumor,
chondrosarcoma, leptomengial disease, diffuse large B-cell
lymphoma, Acute Lymphoblastic Leukemia, Acute Myeloid Leukemia,
Adrenocortical Carcinoma, AIDS-Related Cancers, AIDS-Related
Lymphoma, Anal or Rectal Cancer, Appendix Cancer, Astrocytomas, and
Atypical Teratoid/Rhabdoid Tumor. In one embodiment, the
compositions and methods described herein are used to treat
diseases such as Basal Cell Carcinoma, Basal Cell Nevus Syndrome,
Gorlin-Nevus Syndrome, Bile Duct Cancer, Bladder Cancer, Bone
Cancer, Osteosarcoma and Malignant Fibrous Histiocytoma, Brain
Tumor, Breast Cancer, Bronchial Tumors, Burkitt Lymphoma, and
Spinal Cord Tumors. In one embodiment, the compositions and methods
described herein are used to treat diseases such as Carcinoid
Tumor, Carcinoma of Unknown Primary, Central Nervous System
Atypical Teratoid/Rhabdoid Tumor, Leptomeningeal Disease, Central
Nervous System Embryonal Tumors, Central Nervous System Lymphoma,
Cervical Cancer, Chordoma, Chronic Lymphocytic Leukemia, Chronic
Myelogenous Leukemia, Chronic Myeloproliferative Disorders, Colon
Cancer, Colorectal Cancer, Craniopharyngioma, and Cutaneous T-Cell
Lymphoma (including Sezary syndrome and mycosis fungoides (MF)). In
one embodiment, the compositions and methods described herein are
used to treat cdiseases such as Embryonal Tumors of Central Nervous
System, Endometrial Cancer, Ependymoblastoma, Ependymoma,
Esophageal Cancer, Ewing Sarcoma Family of Tumors, Extracranial
Germ Cell Tumor, Extragonadal Germ Cell Tumor, Extrahepatic Bile
Duct Cancer, and Eye Cancer, including Intraocular Melanoma and
Retinoblastoma. In one embodiment, the compositions and methods
described herein are used to treat diseases such as Gallbladder
Cancer, Gastric (Stomach) Cancer, Gastrointestinal Carcinoid Tumor,
Gastrointestinal Stromal Tumor (GIST), Germ Cell Tumor, Gestational
Trophoblastic Tumor, and Glioma. In one embodiment, the
compositions and methods described herein are used to treat a
cancer selected from the group consisting of Hairy Cell Leukemia,
Head and Neck Cancer, Hepatocellular (Liver) Cancer, Histiocytosis,
Hodgkin Lymphoma, and Hypopharyngeal Cancer. In one embodiment, the
compositions and methods described herein are used to treat
diseases such as Kaposi Sarcoma and Kidney (Renal Cell) Cancer. In
one embodiment, the compositions and methods described herein are
used to treat diseases such as Langerhans Cell Histiocytosis,
Laryngeal Cancer, Lip and Oral 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. In one embodiment, the compositions and methods described
herein are used to treat diseases such as Waldenstrom's
macroglobulinemia (lymphoplasmacytic lymphoma), Malignant Fibrous
Histiocytoma of Bone and Osteosarcoma, Medulloblastoma,
Medulloepithelioma, Melanoma, Merkel Cell Carcinoma, Mesothelioma,
Metastatic Squamous Neck Cancer with Occult Primary, Multiple
Endocrine Neoplasia Syndrome, Mouth Cancer, Multiple Myeloma/Plasma
Cell Neoplasm, Mycosis Fungoides, Myelodysplastic Syndromes,
complex karyotype, blastic phase leukemia,
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 Cavity and Paranasal Sinus Cancer,
Nasopharyngeal Cancer, and Neuroblastoma. In one embodiment, the
compositions and methods described herein are used to treat
diseases such as Oral Cancer, Lip and Oral Cavity Cancer,
Oropharyngeal Cancer, Osteosarcoma and Malignant Fibrous
Histiocytoma of Bone, Ovarian Cancer, Ovarian Germ Cell Tumor,
Ovarian Epithelial Cancer, and Ovarian Low Malignant Potential
Tumor. In one embodiment, the compositions and methods described
herein are used to treat diseases such as Pancreatic Cancer,
Papillomatosis, Paranasal Sinus and Nasal Cavity Cancer,
Parathyroid Cancer, Penile Cancer, Pharyngeal Cancer, Pineal
Parenchymal Tumors of Intermediate Differentiation, Pineoblastoma
and Supratentorial Primitive Neuroectodermal Tumors, Pituitary
Tumor, Pleuropulmonary Blastoma, Pregnancy and Breast Cancer,
Primary Central Nervous System Lymphoma, and Prostate Cancer. In
one embodiment, the compositions and methods described herein are
used to treat a cancer selected from the group consisting of Rectal
Cancer, Renal Cell (Kidney) Cancer, Renal Pelvis and Ureter,
Respiratory Tract Carcinoma Involving the NUT Gene on Chromosome
15, Retinoblastoma, and Rhabdomyosarcoma. In one embodiment, the
compositions and methods described herein are used to treat high
grade prostate cancer. In one embodiment, the compositions and
methods described herein are used to treat 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. In one embodiment,
the compositions and methods described herein are used to treat a
nervous system tumor. In one embodiment, the compositions and
methods described herein are used to treat a central nervous system
tumor. In one embodiment, the compositions and methods described
herein are used to treat a peripheral nervous system tumor. In one
embodiment, the compositions and methods described herein are used
to treat a paraganglioma. In one embodiment, the compositions and
methods described herein are used to treat a pheochromocytoma.
[0186] In in vitro models, in animal models, and in human clinical
trials compound (1) (ONC201) has broad anti-cancer activity, low
toxicity including few, if any, adverse effects, low genotoxicity,
and high bioavailability including oral bioavailability. These
features allow ONC201 and various analogs to be particularly well
suited for pediatric patients. These features also make ONC201 and
various analogs particularly well suited for chronic therapy, for
high risk patients, and to ensure long-lasting responses or stable
disease or to prevent disease recurrence.
[0187] In one embodiment, the compositions and methods described
herein are used to treat a pediatric cancer (e.g., pediatric solid
tumors, pediatric sarcomas, pediatric Ewing's sarcomas, pediatric
gliomas, pediatric central nervous system cancers, pediatric
neuroblastoma, pediatric leukemia and pediatric lymphoma).
[0188] In one embodiment, the compositions and methods described
herein are used to treat a proliferative skin disorder such as
psoriasis. In one embodiment, the compositions and methods
described herein are used to treat a cancer selected from the group
consisting of Salivary Gland Cancer, Sarcoma, Sezary Syndrome, Skin
Cancer, Ocular Cancer, Skin Carcinoma, Small Intestine Cancer, Soft
Tissue Sarcoma, Squamous Cell Carcinoma, Squamous Neck Cancer with
Occult Primary, and Supratentorial Primitive Neuroectodermal
Tumors. In one embodiment, the compositions and methods described
herein are used to treat a cancer selected from the group
consisting of T-Cell Lymphoma, Testicular Cancer, Throat Cancer,
Thymoma and Thymic Carcinoma, Thyroid Cancer, Transitional Cell
Cancer of the Renal Pelvis and Ureter, and Gestational
Trophoblastic Tumor. In one embodiment, the compositions and
methods described herein are used to treat a cancer selected from
the group consisting of Carcinoma of Unknown Primary Site, Cancer
of Unknown Primary Site, Unusual Cancers of Childhood, Transitional
Cell Cancer of the Renal Pelvis and Ureter, Urethral Cancer, and
Uterine Sarcoma. In one embodiment, the compositions and methods
described herein are used to treat cancer selected from the group
consisting of Vaginal Cancer and Vulvar Cancer. In one embodiment,
the compositions and methods described herein are used to treat a
cancer selected from the group consisting of Wilms Tumor and
Women's Cancers.
[0189] In one embodiment, the compositions and methods described
herein are used as a first-line therapy (sometimes called primary
therapy). In one embodiment, the compositions and methods described
herein are used as a second-line therapy. In one embodiment, the
compositions and methods described herein are used as a third-line
therapy. In one embodiment, the compositions and methods described
herein are used as a salvage therapy. The term "salvage therapy"
means a therapeutic agent that can be taken with any regimen after
a subject's initial treatment regimen has failed or after the
subject's condition has not responded to an initial treatment. In
one embodiment, the compositions and methods described herein are
used as a rescue therapy. In one embodiment of the rescue therapy,
the compositions are used as a rescue agent to counteract the
action of an initial treatment. In one embodiment of the rescue
therapy, the compositions are used as rescue agent which is
administered to a subject who has developed resistance to a
standard or an initial treatment. In one embodiment, the
compositions and methods described herein are used as a neoadjuvant
therapy. In one embodiment, the neoadjuvant therapy comprises
administration of one or more of the therapeutic agents described
herein to a subject before a main or first line treatment. In one
embodiment, the neoadjuvant therapy reduces the size or extent of
the cancer being treated before a main or first line treatment is
administered to the subject undergoing treatment. In one
embodiment, the compositions and methods described herein are used
as an adjuvant therapy. In one embodiment, the adjuvant therapy
comprises administration of one or more therapeutic agents
described herein to a subject, wherein the one or more therapeutic
agent that modify the effect of other therapeutic agents that are
already administered to the subject or are concurrently
administered to the subject or subsequently administered to the
subject.
[0190] In one embodiment, the compositions and methods described
herein exhibit reduced chance of drug-drug interactions. In one
embodiment, an imipridone, such as ONC201, or an analog thereof are
eliminated from the patient's body before it can interact with
another pharmaceutically active agent.
[0191] In one embodiment, the compositions and methods of described
herein exhibit toxicity levels that facilitates combinations with
other pharmaceutical agents.
[0192] The methods and compositions described herein are not
limited to a particular animal species. In one embodiment, a
subject treated according to methods and using compositions
described herein, can be mammalian or non-mammalian. In one
embodiment, a mammalian subject mammal includes, but is not limited
to, a human; a non-human primate; a rodent such as a mouse, rat, or
guinea pig; a domesticated pet such as a cat or dog; a horse, cow,
pig, sheep, goat, or rabbit. In one embodiment, a non-mammalian
subject includes, but is not limited to, a bird such as a duck,
goose, chicken, or turkey. In one embodiment, the subject is a
human. In one embodiment, subjects can be either gender and any
age. The composition and methods can also be used to prevent
cancer. The composition and methods can also be used to stimulate
the immune system.
[0193] The methods and compositions described herein are not
limited to a particular age of the subject. In one embodiment, a
subject treated according to methods and using compositions
described herein is over 50 years old, over 55 years old, over 60
years old, or over 65 years old. In one embodiment, a subject
treated according to methods and using compositions described
herein is under 50 years old, under 55 years old, under 60 years
old, or under 65 years old.
[0194] In one embodiment, a subject treated according to methods
and using compositions described herein is a pediatric patient. In
one embodiment, the pediatric patient is younger than 18 years old,
younger than 17 years old, younger than 16 years old, younger than
15 years old, younger than 14 years old, is younger than 13 years
old, younger than 12 years old, younger than 11 years old, younger
than 10 years old, younger than 9 years old, younger than 8 years
old, younger than 7 years old, younger than 6 years old, younger
than 5 years old, younger than 4 years old, younger than 3 years
old, younger than 2 years old, younger than 1 year old. In one
embodiment, the pediatric patient is younger than 12 months old,
younger than 11 months old, younger than 10 months old, younger
than 9 months old, younger than 8 months old, younger than 7 months
old, younger than 6 months old, is younger than 5 months old,
younger than 4 months old, younger than 3 months old, younger than
2 months old, younger than 1 month old. In one embodiment, the
pediatric patient younger than 4 weeks old, younger than 3 weeks
old, younger than 2 weeks old, younger than 1 weeks old. In one
embodiment, the pediatric patient is younger than 7 days old,
younger than 6 days old, younger than 5 days old, younger than 4
days old, younger than 3 days old, younger than 2 days old, or
younger than 1 day old. In one embodiment, the pediatric patient is
a neonate. In one embodiment, the pediatric patient is prematurely
born.
[0195] In one embodiment, the patient is less than 45 kg in weight,
less than 40 kg in weight, less than 35 kg in weight, less than 30
kg in weight, less than 25 kg in weight, less than 20 kg in weight,
less than 15 kg in weight, less than 14 kg in weight, less than 10
kg in weight, less than 5 kg in weight, less than 4 kg in weight,
less than 3 kg in weight, less than 2 kg in weight, or less than 1
kg in weight.
[0196] In one embodiment, the subject has received at least one
prior therapeutic agent. In one embodiment the subject has received
at least two, at least three, or at least four prior therapeutic
agents. In one embodiment the prior therapeutic agent is ibrutinib,
bortezomib, carfilzomib, temozolomide, bevacizumab,
cyclophosphamide, hydroxydaunorubicin, vincristine, prednisone,
cytarabine, cisplatin, rituximab, 5-fluorouracil, oxaliplatin,
leucovorin, or lenalidomide.
[0197] 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 been treated with
adoptive T-cell therapy.
[0198] In one embodiment, the cancer no longer responds to
treatment with ibrutinib, bortezomib, carfilzomib, temozolomide,
bevacizumab, cyclophosphamide, hydroxydaunorubicin, vincristine,
prednisone, cytarabine, cisplatin, rituximab, 5-fluorouracil,
oxaliplatin, leucovorin, lenalidomide, radiation, surgery, or a
combination thereof.
[0199] In one embodiment, the compositions and methods described
herein have a dose response relation in cancer cells that is
different from the dose response relation of the same compositions
and methods in normal cells. The dose response relation of ONC201
on proliferation and cell death in normal and tumor cells was
determined by measuring cell viability following treatment with
ONC201 at various concentrations for 72 hours. The tumors tested
included a human colon cancer cell line (HCT116), breast tumor cell
line (MDA-MB-231), and a human primary glioblastoma cell line
(U87). And the normal cells tested included human foreskin
fibroblasts (HFF), human fetal lung fibroblast (MRC-5) cells, and a
human lung fibroblast cell line (WI-38). Doxorubicin was used as a
positive control at 1 .mu.g/mL in normal fibroblasts. Cell
viability of normal cells tested was at least about 75% at about
1-5 mg/mL of ONC201, whereas viability of tumor cells was
significantly lower (e.g., at or below 50%) at the same ONC201
concentration. Moreover, as ONC201 concentration increased beyond
about 5 mg/mL viability of tumor cells fell to below 25%, whereas
viability of normal cells remained at about 75%. Cell viability
assays in human fetal lung fibroblast (MRC-5) cells were performed
following 72 hour treatment with compound (1) (5 .mu.M) or DMSO and
a recovery period in complete drug-free media after treatment. Cell
recovery was seen with ONC201, but not with DMSO.
[0200] In one embodiment, the compositions and methods described
herein have utility in treating cancer in a subject. In one
embodiment, the compositions and methods described herein have
utility in treating cancer in a human subject. In one embodiment,
the treatment method comprises administering to a subject in need
of such treatment, a pharmaceutically effective amount of an
imipridone, such as ONC201, or an analog thereof, or a
pharmaceutically acceptable salt thereof and a pharmaceutically
acceptable carrier.
[0201] In one embodiment, the treatment method comprises
administering to a subject in need of such treatment: (i) a first
therapeutic agent including an imipridone, such as ONC201, or an
analog thereof, or a pharmaceutically acceptable salt thereof in
combination with (ii) a second therapeutic agent, wherein the first
and the second therapeutic agents are administered either
simultaneously or sequentially. The second therapeutic agent can be
any suitable therapeutic agent, including any pharmaceutically
active agent disclosed herein. A pharmaceutically acceptable ONC201
salt includes the di-hydrochloride salt below:
##STR00017##
[0202] It is understood that a di-hydrochloride salt of ONC201 or
an analog thereof (including a compound of formula (10)), or an
alternative di-salt thereof apparent from the teaching of this
disclosure, can be substituted for ONC201 or an analog thereof in a
composition or dosing regimen described herein.
[0203] In one embodiment, the treatment method comprises
administering a synergistic pharmaceutical combination, either
simultaneously or sequentially, to a subject in need of such
treatment, wherein the synergistic pharmaceutical combination
comprises (i) a first therapeutic agent comprising an imipridone,
such as ONC201, or an analog thereof, or a pharmaceutically
acceptable salt thereof; and (ii) a second therapeutic agent. In
one embodiment, the treatment method comprises administering to a
subject in need of such treatment, either simultaneously or
sequentially, therapeutically synergistic effective amounts of the
first therapeutic agent in combination with the second therapeutic
agent. In one embodiment, the treatment method comprises
administering to a subject in need of such treatment, an effective
amount of the first therapeutic agent in combination with an
effective amount of the second therapeutic agent, wherein the
combination provides a synergistic effect in the in vivo treatment
of a cancer sensitive to the combination, and wherein the first and
the second therapeutic agents are administered either
simultaneously or sequentially. In one embodiment, the treatment
method comprises administering to a subject in need of such
treatment, an effective amount of the first therapeutic agent in
combination with an effective amount of a second therapeutic agent,
wherein the combination provides a synergistic effect in the in
vivo treatment of a minimal residual disease sensitive to the
combination, and wherein the first and second therapeutic agents
are administered either simultaneously or sequentially. In one
embodiment, the second therapeutic agent is given before or prior
to the first therapeutic agent.
[0204] In one embodiment, the treatment method targets a cancer
selected from the group consisting of solid tumors, liquid tumors,
lymphomas, leukemias, or myelomas.
[0205] In one embodiment, the treatment method targets a solid
tumor, wherein the solid tumor is selected from the group
consisting of: Cervical Cancer, Endometrial Cancer, Extracranial
Germ Cell Tumor; Extragonadal Germ Cell Tumor; Germ Cell Tumor;
[0206] Gestational Trophoblastic Tumor; Ovarian Cancer, Ovarian
Germ Cell Tumor, Ovarian Epithelial Cancer, and Ovarian Low
Malignant Potential Tumor; Penile Cancer, Prostate Cancer;
Pregnancy 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 (Liver) Cancer; Kidney (Renal Cell) Cancer; Liver
Cancer, Renal Cell (Kidney) Cancer, Renal Pelvis and Ureter; Basal
Cell Carcinoma; Basal Cell Nevus Syndrome, Gorlin-Nevus Syndrome,
Melanoma, Merkel Cell Carcinoma, Papillomatosis, Multiple Endocrine
Neoplasia Syndrome; Pancreatic Cancer, Parathyroid Cancer, ocular
melanoma; Eye Cancer; Retinoblastoma; Malignant Fibrous
Histiocytoma; Ewing Sarcoma Family of Tumors; desmoplastic round
cell tumor; chondrosarcoma, Kaposi Sarcoma, Rhabdomyosarcoma;
Spinal Cord Tumors, Leptomeningeal Disease, Central Nervous System
Embryonal Tumors, Chordoma, Embryonal Tumors of Central Nervous
System, Ependymoblastoma, Ependymoma, Neuroblastoma; Pineal
Parenchymal Tumors of Intermediate Differentiation, Pineoblastoma;
Adrenocortical Carcinoma; Bone Cancer, Osteosarcoma; Malignant
Fibrous Histiocytoma of Bone and Osteosarcoma; Osteosarcoma and
Malignant Fibrous Histiocytoma of Bone; Carcinoid Tumor, Carcinoma
of Unknown Primary, Bronchial Tumors, Lung Cancer, Pleuropulmonary
Blastoma; Respiratory Tract Carcinoma Involving the NUT Gene on
Chromosome 15, Astrocytomas, Atypical Teratoid/Rhabdoid Tumor;
Central Nervous System Atypical Teratoid/Rhabdoid Tumor,
Craniopharyngioma, Glioma, Brain cancer, Medulloblastoma,
Medulloepithelioma, Supratentorial Primitive Neuroectodermal
Tumors; Pituitary Tumor; Gastric (Stomach) Cancer, Gastrointestinal
Carcinoid Tumor, Gastrointestinal Stromal Tumor (GIST), Bladder
Cancer, Anal or Rectal Cancer, Appendix Cancer, Esophageal Cancer,
Hypopharyngeal Cancer; Laryngeal Cancer, Lip and Oral Cavity
Cancer, Metastatic Squamous Neck Cancer with Occult Primary, Mouth
Cancer, Nasal Cavity and Paranasal Sinus Cancer, Nasopharyngeal
Cancer, Oral Cancer, Lip and Oral Cavity Cancer, Oropharyngeal
Cancer, Paranasal Sinus and Nasal Cavity Cancer, Pharyngeal Cancer;
Head and Neck Cancer, and Mesothelioma.
[0207] In one embodiment, the treatment method targets a lymphoma
selected from the group consisting of: diffuse large B-cell
lymphoma, AIDS-Related Lymphoma, Cutaneous T-Cell Lymphoma, Sezary
syndrome, mycosis fungoides (MF); Histiocytosis; Burkitt Lymphoma,
and Central Nervous System Lymphoma; Non-Hodgkin Lymphoma, and
Primary Central Nervous System Lymphoma, Hodgkin Lymphoma,
Waldenstrom's macroglobulinemia; Mycosis Fungoides; Primary Central
Nervous System Lymphoma; lymphoplasmacytic lymphoma, and Primary
Central Nervous System Lymphoma.
[0208] In one embodiment, the treatment method targets a
Non-Hodgkin's lymphoma (NHL) selected from the group consisting of:
mantle cell lymphoma, diffuse large B-cell lymphoma, follicular
lymphoma, marginal zone lymphoma, small lymphocytic lymphoma,
lyphoplasmacytic NHL, Waldenstrom's macroglobulinaemia, and skin
lymphomas.
[0209] In one embodiment, the treatment method targets a leukemia
selected from the group consisting of: Acute Lymphoblastic Leukemia
(ALL), Chronic Lymphocytic Leukemia (CLL), Chronic
Myeloproliferative Disorders; Hairy Cell Leukemia; Acute Myeloid
Leukemia (AML); Chronic Myelogenous Leukemia (CML); and Langerhans
Cell Histiocytosis.
[0210] In one embodiment, the treatment method targets an acute
leukemia selected from the group consisting of: acute lymphotyte
leukemia, acute myeloid leukemia, chronic lymphoblasitc leukemia,
chronic myeloid leukemia, myelodysplastic syndrome, and
myeloproliferative disease.
[0211] In one embodiment, the treatment method targets a myeloma
selected from the group consisting of: IgA myeloma; IgG myeloma;
IgM myeloma; IgD myeloma; IgE myeloma; light chain myeloma; non
secretory myeloma; complex karyotype, blastic phase leukemia;
Multiple Myeloma/Plasma Cell Neoplasm, Multiple Myeloma,
Myelodysplastic Syndromes, Myelodysplastic/Myeloproliferative
Neoplasms, and Myeloproliferative Disorders.
[0212] In one embodiment, the treatment method targets a peripheral
nervous system tumor. In one embodiment, the treatment method
targets a paraganglioma. In one embodiment, the treatment method
targets a pheochromocytoma.
[0213] In one embodiment, treatment of cancer comprises prevention
of tumor growth in a cancer subject. In one embodiment, treatment
of cancer comprises prevention of formation of cancer metastases in
a cancer subject. In one embodiment, treatment of cancer comprises
targeted treatment of minimal residual disease in a cancer subject
known to have the minimal residual disease in a cancer or a subject
at risk for having minimal residual disease.
[0214] This might be indicated after treatment of the primary tumor
by surgery and/or after chemotherapy (radiotherapy) has been
initiated or determined to be efficaceous.
[0215] Disseminated tumor cells may be in their dormant state and
often cannot be attacked by chemotherapy (radiotherapy). A thus
treated patient seemingly is in a healed state, and referred to as
"minimal residual disease." Nevertheless, the dormant tumor cells
have a potential to form metastases if they become metastasising
cells due to a growth stimulus after a longer dormant state.
[0216] The term "minimal residual disease" denotes a small number
of cancer cells that remain in a subject during or after treatment
when the subject is in remission (exhibiting no symptoms or signs
of the disease). The methods described herein are preferably
applied to a form of the diseases listed herein, including adult
and childhood forms of these diseases.
[0217] In one embodiment, the treatment method is useful for
treating an autoimmune disease. Autoimmune diseases include, but
are not limited to alopecia areata, antiphospholipid, autoimmune
hepatitis, celiac disease, diabetes type 1, Graves' disease,
Guillain-Barre syndrome, Hashimoto's disease, hemolytic anemia,
idiopathic thrombocytopenic purpura, inflammatory bowel disease,
inflammatory myopathies, multiple sclerosis, primary biliary
cirrhosis, psoriasis, rheumatoid arthritis, scleroderma, Sjogren's
syndrome, systemic lupus erythematosus, and vitiligo.
[0218] In one embodiment, the treatment method is useful for
treating autoimmune and inflammatory disorders of the peripheral
nerve system such as amyotrophic lateral sclerosis (Lou Gehrig's
disease), based on various causes such as metabolic disorders that
include diabetes, B12 and folate vitamin deficiencies, chemotherapy
medications and medicines used to treat HIV, poisons that cause
peripheral nerve damage, cancers that develop peripheral
neuropathies as well as paraneoplastic syndromes, alcohol abuse,
chronic kidney disease, injuries that cause compression on nerves
and other lesions, infections such as Lyme disease, Guillain Barre
syndrome, connective tissue disease, rheumatoid arthritis,
Sjogren's syndrome, systemic lupus erythematosus, certain
inflammatory conditions such as sarcoidosis, coeliac disease,
hereditary diseases such as charcot marie tooth syndrome,
Friedreich's ataxia, and/or idiopathic where no specific cause is
found but inflammatory and/or autoimmune mechanisms are the cause
of onset.
[0219] In one embodiment, the treatment method is useful for
treating autoimmune and inflammatory disorders with ocular
manifestations. Such ocular manifestations include, but are not
limited to, ocular cicatricial pemphigoid, Mooren's corneal ulcer,
various forms of uveitis, rheumatoid arthritis, systemic lupus
erythematosus, polyarteritis nodosa, relapsing polychondritis,
Wegener's granulomatosis, scleroderma, Behcet's disease, Reiter's
disease, inflammatory bowel disease (ulcerative colitis and Crohn's
disease) and ankylosing spondylitis, retinitis pigmentosa, macular
degeneration, keratoconjunctivitis sicca, scleritis, episcleritis,
keratitis, peripheral corneal ulceration, and less common entities
such as choroiditis, retinal vasculitis, episcleral nodules,
retinal detachments, and/or macular edema.
[0220] In one embodiment, the treatment method is useful for
treating acute allograft rejection in transplant patients. In one
embodiment, the treatment method is useful for treating ischemic
stroke. In one embodiment, the treatment method is useful for
treating inflammatory diseases including arthritis, psoriasis,
asthma, and colitis.
[0221] In one embodiment, a therapeutic agent includes a
pharmaceutically acceptable mono-salt of ONC201 or an analog
thereof (e.g., a compound of formula (10)). In one embodiment, a
therapeutic agent includes a pharmaceutically acceptable ONC201
di-salt or an analog thereof (e.g., a compound of formula (10)). As
described herein, some of the analogs can be tri-salts In one
embodiment, a therapeutic agent includes ONC201 or an analog
thereof (e.g., a compound of formula (10)) in the form of a
pharmaceutically acceptable mono- or di-salt selected from the
group consisting of hydrochloride, hydrobromide, hydrogensulphate,
sulfates, phosphates, fumarates, succinates, oxalates and lactates,
bisulfates, hydroxyl, tartrate, nitrate, citrate, bitartrate,
carbonate, malate, maleate, fumarate sulfonate, methylsulfonate,
formate, acetate, and carboxylate. In one embodiment, a therapeutic
agent includes ONC201 or an analog thereof in the form of a
pharmaceutically acceptable mono- or di-salt selected from
p-toluene-sulfonate, benzenesulfonate, methanesulfonate, oxalate,
succinate, tartrate, citrate, fumarate and maleate. In one
embodiment, a therapeutic agent includes ONC201 or an analog
thereof in the form of a pharmaceutically acceptable mono- or
di-salt having a counter ion selected from the group consisting of
ammonium, sodium, potassium, calcium, magnesium, zinc, lithium,
and/or with counter-ions such as methylamino, dimethylamino,
diethylamino, triethylamino counter-ions, and combinations thereof.
In one embodiment, a therapeutic agent includes a compound
described herein in the form of a halide di-salt, such as a
di-hydrochloride salt or a di-hydrobromide salt.
[0222] In one embodiment of the treatment method, the second
therapeutic agent includes an anti-cancer agent. In one embodiment
of the treatment method, the second therapeutic agent is selected
from acivicin, aclarubicin, acodazole, acronine, adozelesin,
aldesleukin, alitretinoin, allopurinol, altretamine, ambomycin,
ametantrone, amifostine, aminoglutethimide, amsacrine, anastrozole,
anthramycin, arsenic trioxide, asparaginase, asperlin, azacitidine,
azetepa, azotomycin, batimastat, benzodepa, bevacizumab,
bicalutamide, bisantrene, bisnafide dimesylate, bizelesin,
bleomycin, brequinar, bropirimine, busulfan, cactinomycin,
calusterone, capecitabine, caracemide, carbetimer, carboplatin,
carmustine, carubicin, carzelesin, cedefingol, celecoxib,
chlorambucil, cirolemycin, cisplatin, cladribine, crisnatol
mesylate, cyclophosphamide, cytarabine, dacarbazine, dactinomycin,
daunorubicin, decitabine, dexormaplatin, dezaguanine, dezaguanine
mesylate, diaziquone, docetaxel, doxorubicin, droloxifene,
dromostanolone, duazomycin, edatrexate, eflomithine, elsamitrucin,
enloplatin, enpromate, epipropidine, epirubicin, erbulozole,
esorubicin, estramustine, etanidazole, etoposide, etoprine,
fadrozole, fazarabine, fenretinide, floxuridine, fludarabine,
fluorouracil, flurocitabine, fosquidone, fostriecin, fulvestrant,
gemcitabine, hydroxyurea, idarubicin, ifosfamide, ilmofosine,
interleukin II (IL-2, including recombinant interleukin II or
rIL2), interferon .alpha.-2a, interferon .alpha.-2b, interferon
.alpha.-n1, interferon .alpha.-n3, interferon .beta.-Ia, interferon
gamma-Ib, iproplatin, irinotecan, lanreotide, letrozole,
leuprolide, liarozole, lometrexol, lomustine, losoxantrone,
masoprocol, maytansine, mechlorethamine hydrochlride, megestrol,
melengestrol acetate, melphalan, menogaril, mercaptopurine,
methotrexate, metoprine, meturedepa, mitindomide, mitocarcin,
mitocromin, mitogillin, mitomalcin, mitomycin, mitosper, mitotane,
mitoxantrone, mycophenolic acid, nelarabine, nocodazole,
nogalamycin, ormnaplatin, oxisuran, paclitaxel, pegaspargase,
peliomycin, pentamustine, peplomycin, perfosfamide, pipobroman,
piposulfan, piroxantrone hydrochloride, plicamycin, plomestane,
porfimer, porfiromycin, prednimustine, procarbazine, puromycin,
pyrazofurin, riboprine, rogletimide, safingol, semustine,
simtrazene, sparfosate, sparsomycin, spirogermanium, spiromustine,
spiroplatin, streptonigrin, streptozocin, sulofenur, talisomycin,
tamoxifen, tecogalan, tegafur, teloxantrone, temoporfin,
teniposide, teroxirone, testolactone, thiamiprine, thioguanine,
thiotepa, tiazofurin, tirapazamine, topotecan, toremifene,
trestolone, triciribine, trimetrexate, triptorelin, tubulozole,
uracil mustard, uredepa, vapreotide, verteporfin, vinblastine,
vincristine sulfate, vindesine, vinepidine, vinglycinate,
vinleurosine, vinorelbine, vinrosidine, vinzolidine, vorozole,
zeniplatin, zinostatin, zoledronate, zorubicin and combinations
thereof.
[0223] In one embodiment of the treatment method, the second
therapeutic agent is selected, from hormone analogs and
antihormones, aromatase inhibitors, LHRH agonists and antagonists,
inhibitors of growth factors, growth factor antibodies, growth
factor receptor antibodies, tyrosine kinase inhibitors;
antimetabolites; antitumour antibiotics; platinum derivatives;
alkylation agents; antimitotic agents; tubuline inhibitors; PARP
inhibitors, topoisomerase inhibitors, serine/threonine kinase
inhibitors, tyrosine kinase inhibitors, protein interaction
inhibitors, MEK inhibitors, ERK inhibitors, IGF-1R inhibitors, ErbB
receptor inhibitors, rapamycin analogs, amifostin, anagrelid,
clodronat, filgrastin, interferon, interferon .alpha., leucovorin,
rituximab, procarbazine, levamisole, mesna, mitotane, pamidronate
and porfimer, 2-chlorodesoxyadenosine, 2-fluorodesoxy-cytidine,
2-methoxyoestradiol, 2C4,3-alethine, 131-1-TM-601, 3CPA,
7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A 105972, A
204197, abiraterone, aldesleukin, alitretinoin, allovectin-7,
altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037,
AP-5280, apaziquone, apomine, aranose, arglabin, arzoxifene,
atamestane, atrasentan, auristatin PE, ABT-199 (Venetoclax),
ABT-263 (Navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab),
ARRY 162, ARRY 438162, ARRY-300, ARRY-142886/AZD-6244
(selumetinib), ARRY-704/AZD-8330, AR-12, AR-42, AS-703988,
AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026
(primasertib), avastin, AZD-2014, azacytidine, azaepothilone B,
azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576,
bevacizumab, BEZ-235, biricodar dicitrate, BCX-1777, BKM-120,
bleocin, 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, bleomycinic acid, bleomycin
A, bleomycin B, brivanib, bryostatin-1, bortezomib, brostallicin,
busulphan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol,
canertinib, canfosfamide, capecitabine, carboxyphthalatoplatin,
CC1-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime,
ceflatonin, ceftriaxone, celecoxib, celmoleukin, cemadotin,
CH.sub.4987655/RO-4987655, chlorotrianisene, cilengitide,
ciclosporin, CDA-II, CDC-394, CKD-602, CKI-27, clofarabin,
colchicin, combretastatin A4, COT inhibitors, CHS-828, CH-5132799,
CLL-Thera, CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal
antibodies, CP-461, CV-247, cyanomorpholinodoxorubicin, cytarabine,
D 24851, decitabine, deoxorubicin, deoxyrubicin, deoxycoformycin,
depsipeptide, desoxyepothilone B, dexamethasone, dexrazoxanet,
diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10,
doranidazole, DS-7423, E7010, E-6201, edatrexat, edotreotide,
efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509,
enzastaurin, enzalutamide, elsamitrucin, epothilone B, epratuzumab,
ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine,
ethynyloestradiol, exatecan, exatecan mesylate, exemestane,
exisulind, fenretinide, figitumumab, floxuridine, folic acid,
FOLFOX, FOLFOX4, FOLFIRI, formestane, fotemustine, galarubicin,
gallium maltolate, gefinitib, gemtuzumab, gimatecan, glufosfamide,
GCS-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032,
GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100,
gp100-peptide vaccines, GSK-5126766, GSK-690693, GSK-1120212
(trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A,
GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron,
herceptin, hexamethylmelamine, histamine, homoharringtonine,
hyaluronic acid, hydroxyurea, hydroxyprogesterone caproate,
ibandronate, ibritumomab, idatrexate, idenestrol, IDN-5109, IGF-1R
inhibitors, IMC-1C11, IMC-A12 (cixutumumab), immunol, indisulam,
interferon .alpha.-2a, interferon .alpha.-2b, pegylated interferon
.alpha.-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib,
ipilimumab, iproplatin, irofulven, isohomohalichondrin-B,
isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088,
conjugated oestrogens, kahalid F, ketoconazole, KW-2170, KW-2450,
lobaplatin, leflunomide, lenograstim, leuprolide, leuporelin,
lexidronam, LGD-1550, linezolid, lutetium texaphyrin, lometrexol,
losoxantrone, LU 223651, lurtotecan, LY-S6AKT1, LY-2780301,
mafosfamide, marimastat, mechloroethamine, MEK inhibitors, MEK-162,
methyltestosteron, 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, norelin, N-acetylcysteine, 06-benzylguanine,
oblimersen, omeprazole, oncophage, oncoVEXGM-CSF, ormiplatin,
ortataxel, OX44 antibodies, OSI-027, OSI-906 (linsitinib), 4-1BB
antibodies, oxantrazole, oestrogen, panitumumab, patupilone,
pegfilgrastim, PCK-3145, pegfilgrastim, PBI-1402, PBI-05204,
PD0325901, PD-1 antibodies, 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 inhibitors, PI3K/mTOR
inhibitors, PG-TXL, PG2, PLX-4032/RO-5185426 (vemurafenib),
PLX-3603/RO-5212054, PT-100, PWT-33597, PX-866, picoplatin,
pivaloyloxymethylbutyrate, pixantrone, phenoxodiol O, PKI166,
plevitrexed, plicamycin, polyprenic acid, porfiromycin, prednisone,
prednisolone, quinamed, quinupristin, R115777, RAF-265, ramosetron,
ranpirnase, RDEA-119/BAY 869766, RDEA-436, rebeccamycin analogs,
receptor tyrosine kinase (RTK) inhibitors, revimid, RG-7167,
RG-7304, RG-7421, RG-7321, RG 7440, rhizoxin, rhu-MAb, rinfabate,
risedronate, rituximab, robatumumab, rofecoxib, RO-31-7453,
RO-5126766, RO-5068760, RPR 109881A, rubidazone, rubitecan,
R-flurbiprofen, RX-0201, S-9788, sabarubicin, SAHA, sargramostim,
satraplatin, SB 408075, Se-015/Ve-015, SU5416, SU6668, SDX-101,
semustin, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897,
SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine,
suberanilohydroxamic acid, sutent, T 900607, T 138067, TAK-733,
TAS-103, tacedinaline, talaporf in, Tarceva, tariquitar, tasisulam,
taxotere, taxoprexin, tazarotene, tegafur, temozolamide,
tesmilifene, testosterone, testosterone propionate, tesmilifene,
tetraplatin, tetrodotoxin, tezacitabine, thalidomide, theralux,
therarubicin, thymalfasin, thymectacin, tiazofurin, tipifarnib,
tirapazamine, tocladesine, tomudex, toremofin, trabectedin,
TransMID-107, transretinic acid, traszutumab, tremelimumab,
tretinoin, triacetyluridine, triapine, triciribine, trimetrexate,
TLK-286TXD 258, tykerb/tyverb, urocidin, valrubicin, vatalanib,
vincristine, vinflunine, 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.
[0224] In one embodiment of the treatment method, the second
therapeutic agent is selected from tamoxifen, toremifene,
raloxifene, fulvestrant, megestrol acetate, flutamide, nilutamide,
bicalutamide, aminoglutethimide, cyproterone acetate, finasteride,
buserelin acetate, fludrocortisone, fluoxymesterone,
medroxy-progesterone, octreotide, and combinations thereof. In one
embodiment of the treatment method, the second therapeutic agent is
selected from LHRH agonists and LHRH antagonists. In one
embodiment, a LHRH agonist is selected from goserelin acetate,
luprolide acetate, triptorelin pamoate and combinations thereof. In
one embodiment, the second therapeutic agent includes a LHRH
antagonist is selected from Degarelix, Cetrorelix, Abarelix,
Ozarelix, Degarelix combinations thereof. In one embodiment of the
treatment method, the second therapeutic agent includes an
inhibitor of a growth factor. In one embodiment, the inhibitor of a
growth factor is selected from inhibitors of: platelet derived
growth factor (PDGF), fibroblast growth factor (FGF), vascular
endothelial growth factor (VEGF), epidermal growth factor (EGF),
insuline-like growth factors (IGF), human epidermal growth factor
(HER), hepatocyte growth factor (HGF), and combinations thereof. In
one embodiment, the human epidermal growth factor (HER) is selected
from HER2, HER3, and HER4.
[0225] In one embodiment of the treatment method, the second
therapeutic agent includes a tyrosine kinase inhibitor. In one
embodiment of the treatment method, the tyrosine kinase inhibitor
is selected from cetuximab, gefitinib, imatinib, lapatinib and
trastuzumab, and combinations thereof. In one embodiment of the
treatment method, the second therapeutic agent includes an
aromatase inhibitor. In one embodiment of the treatment method, the
aromatase inhibitor is selected from anastrozole, letrozole,
liarozole, vorozole, exemestane, atamestane, and combinations
thereof.
[0226] In one embodiment of the treatment method, the second
therapeutic agent includes an antimetabolite. In one embodiment of
the treatment method, the antimetabolite comprises an antifolate.
In one embodiment of the treatment method, the antifolate is
selected from methotrexate, raltitrexed, pyrimidine analogs, and
combinations thereof. In one embodiment of the treatment method,
the antimetabolite is a pyrimidine analog. In one embodiment of the
treatment method, the pyrimidine analog is selected from
5-fluorouracil, capecitabin, gemcitabin, and combination thereof.
In one embodiment of the treatment method, the antimetabolite is a
purine analog or an adenosine analog. In one embodiment of the
treatment method, the purine analog or adenosine analog is selected
from mercaptopurine, thioguanine, cladribine and pentostatin,
cytarabine, fludarabine, and combinations thereof. In one
embodiment of the treatment method, the second therapeutic agent
includes an antitumour antibiotic. In one embodiment of the
treatment method, the antitumor antibiotic is selected from
anthracyclins, doxorubicin, daunorubicin, epirubicin and
idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin,
streptozocin and combinations thereof. In one embodiment of the
treatment method, the second therapeutic agent includes a platinum
derivative. In one embodiment of the treatment method, the platinum
derivative is selected from cisplatin, oxaliplatin, carboplatin and
combinations thereof. In one embodiment of the treatment method,
the second therapeutic agent includes an alkylation agent. In one
embodiment of the treatment method, the alkylation agent is
selected from estramustin, meclorethamine, melphalan, chlorambucil,
busulphan, dacarbazin, cyclophosphamide, ifosfamide, temozolomide,
nitrosoureas, and combinations thereof. In one embodiment of the
treatment method, the second therapeutic agent includes a
nitrosourea. In one embodiment of the treatment method, the
nitrosourea is selected from carmustin, lomustin, thiotepa, and
combinations thereof. In one embodiment of the treatment method,
the second therapeutic agent includes an antimitotic agent. In one
embodiment of the treatment method, the antimitotic agent is
selected from Vinca alkaloids and taxanes. In one embodiment of the
treatment method, the taxane is selected from paclitaxel,
docetaxel, and combinations thereof. In one embodiment of the
treatment method, the Vinca alkaloids are selected from
vinblastine, vindesin, vinorelbin, vincristine, and combinations
thereof. In one embodiment of the treatment method, the second
therapeutic agent includes a topoisomerase inhibitor. In one
embodiment of the treatment method, the topoisomerase inhibitor is
an epipodophyllotoxin. In one embodiment of the treatment method,
the topoisomerase inhibitor, which is an epipodophyllotoxin
selected from etoposide, etopophos, teniposide, amsacrin,
topotecan, irinotecan, mitoxantron, and combinations thereof. In
one embodiment of the treatment method, the second therapeutic
agent includes a serine/threonine kinase inhibitor. In one
embodiment of the treatment method, the serine/threonine kinase
inhibitor is 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,
inhibitors of CDKs, Aurora kinase inhibitors, and combinations
thereof. In one embodiment of the treatment method, the second
therapeutic agent includes a tyrosine kinase inhibitor. In one
embodiment of the treatment method, the second therapeutic agent
includes a PTK2/FAK inhibitor. In one embodiment of the treatment
method, the second therapeutic agent includes a protein interaction
inhibitor. In one embodiment of the treatment method, the protein
interaction inhibitor is selected from IAP, Mcl-1, MDM2/MDMX and
combinations thereof. In one embodiment of the treatment method,
the second therapeutic agent includes a rapamycin analog. In one
embodiment of the treatment method, the rapamycin analog is
selected from everolimus, temsirolimus, ridaforolimus, sirolimus,
and combinations thereof. In one embodiment of the treatment
method, the second therapeutic agent is selected from amifostin,
anagrelid, clodronat, filgrastin, interferon, interferon .alpha.,
leucovorin, rituximab, procarbazine, levamisole, mesna, mitotane,
pamidronate and porfimer, and combinations thereof. In one
embodiment of the treatment method, the second therapeutic agent is
selected from 2-chlorodesoxyadenosine, 2-fluorodesoxy-cytidine,
2-methoxyoestradiol, 2C4,3-alethine, 131-1-TM-601, 3CPA,
7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, A 105972, A
204197, abiraterone, aldesleukin, alitretinoin, allovectin-7,
altretamine, alvocidib, amonafide, anthrapyrazole, AG-2037,
AP-5280, apaziquone, apomine, aranose, arglabin, arzoxifene,
atamestane, atrasentan, auristatin PE, ABT-199 (Venetoclax),
ABT-263 (Navitoclax), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitumab),
ARRY 162, ARRY 438162, ARRY-300, ARRY-142886/AZD-6244
(selumetinib), ARRY-704/AZD-8330, AR-12, AR-42, AS-703988,
AXL-1717, AZD-8055, AZD-5363, AZD-6244, ARQ-736, ARQ 680, AS-703026
(primasertib), avastin, AZD-2014, azacytidine, azaepothilone B,
azonafide, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576,
bevacizumab, BEZ-235, biricodar dicitrate, BCX-1777, BKM-120,
bleocin, 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, bleomycinic acid, bleomycin
A, bleomycin B, brivanib, bryostatin-1, bortezomib, brostallicin,
busulphan, BYL-719, CA-4 prodrug, CA-4, CapCell, calcitriol,
canertinib, canfosfamide, capecitabine, carboxyphthalatoplatin,
CC1-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1 cefixime,
ceflatonin, ceftriaxone, celecoxib, celmoleukin, cemadotin,
CH4987655/RO-4987655, chlorotrianisene, cilengitide, ciclosporin,
CDA-II, CDC-394, CKD-602, CKI-27, clofarabin, colchicin,
combretastatin A4, COT inhibitors, CHS-828, CH-5132799, CLL-Thera,
CMT-3 cryptophycin 52, CTP-37, CTLA-4 monoclonal antibodies,
CP-461, CV-247, cyanomorpholinodoxorubicin, cytarabine, D 24851,
decitabine, deoxorubicin, deoxyrubicin, deoxycoformycin,
depsipeptide, desoxyepothilone B, dexamethasone, dexrazoxanet,
diethylstilbestrol, diflomotecan, didox, DMDC, dolastatin 10,
doranidazole, DS-7423, E7010, E-6201, edatrexat, edotreotide,
efaproxiral, eflornithine, EGFR inhibitors, EKB-569, EKB-509,
enzastaurin, enzalutamide, elsamitrucin, epothilone B, epratuzumab,
ER-86526, erlotinib, ET-18-0CH3, ethynylcytidine,
ethynyloestradiol, exatecan, exatecan mesylate, exemestane,
exisulind, fenretinide, figitumumab, floxuridine, folic acid,
FOLFOX, FOLFOX4, FOLFIRI, formestane, fotemustine, galarubicin,
gallium maltolate, gefinitib, gemtuzumab, gimatecan, glufosfamide,
GCS-100, GDC-0623, GDC-0941 (pictrelisib), GDC-0980, GDC-0032,
GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GPX-100,
gp100-peptide vaccines, GSK-5126766, GSK-690693, GSK-1120212
(trametinib), GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A,
GSK-2334470, GSK-2110183, GSK-2141795, GW2016, granisetron,
herceptine, hexamethylmelamine, histamine, homoharringtonine,
hyaluronic acid, hydroxyurea, hydroxyprogesterone caproate,
ibandronate, ibritumomab, idatrexate, idenestrol, IDN-5109, IGF-1R
inhibitors, IMC-1C11, IMC-A12 (cixutumumab), immunol, indisulam,
interferon .alpha.-2a, interferon .alpha.-2b, pegylated interferon
.alpha.-2b, interleukin-2, INK-1117, INK-128, INSM-18, ionafarnib,
ipilimumab, iproplatin, irofulven, isohomohalichondrin-B,
isoflavone, isotretinoin, ixabepilone, JRX-2, JSF-154, J-107088,
conjugated oestrogens, kahalid F, ketoconazole, KW-2170, KW-2450,
lobaplatin, leflunomide, lenograstim, leuprolide, leuporelin,
lexidronam, LGD-1550, linezolid, lutetium texaphyrin, lometrexol,
losoxantrone, LU 223651, lurtotecan, LY-S6AKT1, LY-2780301,
mafosfamide, marimastat, mechloroethamine, MEK inhibitors, MEK-162,
methyltestosteron, 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, norelin, N-acetylcysteine, 06-benzylguanine,
oblimersen, omeprazole, oncophage, oncoVEXGM-CSF, ormiplatin,
ortataxel, OX44 antibodies, OSI-027, OSI-906 (linsitinib), 4-1BB
antibodies, oxantrazole, oestrogen, panitumumab, patupilone,
pegfilgrastim, PCK-3145, pegfilgrastim, PBI-1402, PBI-05204,
PD0325901, PD-1 antibodies, 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 inhibitors, PI3K/mTOR
inhibitors, PG-TXL, PG2, PLX-4032/RO-5185426 (vemurafenib),
PLX-3603/RO-5212054, PT-100, PWT-33597, PX-866, picoplatin,
pivaloyloxymethylbutyrate, pixantrone, phenoxodiol O, PKI166,
plevitrexed, plicamycin, polyprenic acid, porfiromycin, prednisone,
prednisolone, quinamed, quinupristin, R115777, RAF-265, ramosetron,
ranpirnase, RDEA-119/BAY 869766, RDEA-436, rebeccamycin analogs,
receptor tyrosine kinase (RTK) inhibitors, revimid, RG-7167,
RG-7304, RG-7421, RG-7321, RG 7440, rhizoxin, rhu-MAb, rinfabate,
risedronate, rituximab, robatumumab, rofecoxib, RO-31-7453,
RO-5126766, RO-5068760, RPR 109881A, rubidazone, rubitecan,
R-flurbiprofen, RX-0201, S-9788, sabarubicin, SAHA, sargramostim,
satraplatin, SB 408075, Se-015/Ve-015, SU5416, SU6668, SDX-101,
semustin, seocalcitol, SM-11355, SN-38, SN-4071, SR-27897,
SR-31747, SR-13668, SRL-172, sorafenib, spiroplatin, squalamine,
suberanilohydroxamic acid, sutent, T 900607, T 138067, TAK-733,
TAS-103, tacedinaline, talaporf in, Tarceva, tariquitar, tasisulam,
taxotere, taxoprexin, tazarotene, tegafur, temozolamide,
tesmilifene, testosterone, testosterone propionate, tesmilifene,
tetraplatin, tetrodotoxin, tezacitabine, thalidomide, theralux,
therarubicin, thymalfasin, thymectacin, tiazofurin, tipifarnib,
tirapazamine, tocladesine, tomudex, toremofin, trabectedin,
TransMID-107, transretinic acid, traszutumab, tremelimumab,
tretinoin, triacetyluridine, triapine, triciribine, trimetrexate,
TLK-286TXD 258, tykerb/tyverb, urocidin, valrubicin, vatalanib,
vincristine, vinflunine, 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.
[0227] In one embodiment, the other therapeutic agent comprises a
steroid, including dexamethasone, prednisolone, methyl
prednisolone, prednisone, hydrocortisone, triamcinolone,
betamethasone, and cortivazol. In one embodiment, the other
therapeutic agent comprises an anti-emetic. Anti-emetics include,
but are not limited to, 5-HT3 receptor agonists (such as
dolasetron, granisetron, ondansetron, tropisetron, palonosetron,
and mirtazapine), dopamine agonists (such as domperidone,
olanzapine, droperidol, haloperidol, chlorpromazine,
prochlorperazine, alizapride, prochlorperazine, and
metoclopramide), NK1 receptor antagonists (such as aprepitant and
casopitant), antihistamines (such as cyclizine, diphenhydramine,
dimenhydrinate, doxylamine, meclizine, promethazine, hydroxyzine),
cannabinoids (such as cannabis, dronabinol, nabilone, and sativex),
benzodiazepines (such as midazolam and lorazepam), anticholinergics
(such as hyoscine), trimethobenzamide, ginger, emetrol, propofol,
peppermint, muscimol, and ajwain.
[0228] Pharmaceutical compositions may be administered to a subject
via any suitable route of administration. In one embodiment, the
pharmaceutical composition is administered to a subject orally,
parenterally, transdermally or transmucosally. In one embodiment,
the pharmaceutical composition is administered to a subject
parenterally. In one embodiment, the pharmaceutical composition is
administered to a subject via a parenteral route of administration
selected from the group consisting of intravenous (IV),
subcutaneous (SC), and intramuscular (IM). In one embodiment, the
pharmaceutical composition is administered to a subject via a route
of administration selected from rectal and transdermal. In one
embodiment, the pharmaceutical composition is administered to a
subject in a dosage form selected from the group consisting of
sterile solutions, suspensions, suppositories, tablets and
capsules. In one embodiment, the pharmaceutical composition is
administered to a subject in an oral dosage form selected from the
group consisting of a tablet, caplet, capsule, lozenge, syrup,
liquid, suspension and elixir. In one embodiment, the
pharmaceutical composition is administered to a subject in an oral
dosage form selected from the group consisting of tablets, hard
shell capsules, soft gelatin capsules, beads, granules, aggregates,
powders, gels, solids and semi-solids.
[0229] In one embodiment, the pharmaceutical composition is
administered to a subject as a dosage form selected from the group
consisting of sustained release forms, controlled release forms,
delayed release forms and response release forms.
[0230] In one embodiment, the pharmaceutical composition is
administered to a subject once daily. In one embodiment, the
pharmaceutical composition is administered to a subject according
to an infrequent dosing regimen (e.g., administered once per week
or less frequently). In one embodiment, the pharmaceutical
composition is administered to a subject according to a frequent
dosing regimen (e.g., administered more than once per week). In one
embodiment, the pharmaceutical composition is administered to a
subject once weekly. In one embodiment, the pharmaceutical
composition is administered to a subject once every four weeks. In
one embodiment, the pharmaceutical composition is administered to a
subject twice a week. In one embodiment, the pharmaceutical
composition is administered to a subject once every two weeks. In
one embodiment, the pharmaceutical composition is administered to a
subject once every three weeks. In one embodiment, the
pharmaceutical composition is administered to a subject in a
repeated cycle of once weekly, once every two weeks, once every
three weeks, once every four weeks or combinations thereof.
[0231] In one embodiment, the treatment method comprises
administering to a subject in need of such treatment: (i) a first
therapeutic agent including a compound comprising an imipridone,
such as ONC201, or an analog thereof, or a pharmaceutically
acceptable salt thereof in combination with (ii) a second
therapeutic agent, wherein the first therapeutic agent and the
second therapeutic agent are administered either simultaneously or
sequentially; and further comprises assaying the expression of an
endoplasmic reticulum (ER) stress response gene in a biological
sample. In one embodiment, the endoplasmic reticulum stress
response gene is selected from the group that includes, but is not
limited to, C/EBP-Homologous Protein (CHOP), Activating
Transcription Factor 3 (ATF3) and both CHOP and ATF3. In one
embodiment, the endoplasmic reticulum stress response gene is
selected from the group that includes, but is not limited to, ATF3,
Activating Transcription Factor 4 (ATF4) CHOP, IRE1, Binding
immunoglobulin protein (BiP), Eukaryotic translation initiation
factor 2A (eIF2a), X-box binding protein 1 (XBP1). The biological
sample may be tumor, peripheral blood mononuclear cells, or skin
biopsy. The biological sample may be obtained before, during, or
after drug administration. In one embodiment, the treatment method
further comprises adjusting a dose of the first therapeutic agent
to achieve induction of about 50%, 75%, 100%, 125%, 150%, 175%,
200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%,
475%, 500%, 525%, 550%, 575%, 600%, or greater than 600% of one or
more ER stress gene. In one embodiment, the treatment method
further comprises adjusting a 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% to about 250%,
about 250% to about 300%, about 300% to about 350%, about 350% to
about 400%, about 400% to about 450%, about 450% to about 500%,
about 500% to about 550%, about 550% to about 600%, or greater than
600% of ER stress genes. In one embodiment, the treatment method
further comprises adjusting a dose of the first therapeutic agent
to achieve induction of about 50% to about 100%, about 100% to
about 200%, about 200% to about 300%, about 300% to about 400%,
about 400% to about 500%, about 500% to about 600%, or greater than
600% of ER stress genes.
[0232] In one embodiment, the treatment method comprises
administering to a subject in need of such treatment: (i) a first
therapeutic agent including a compound comprising an imipridone,
such as ONC201, an analog thereof, or a pharmaceutically acceptable
salt thereof in combination with (ii) a second therapeutic agent,
wherein the first therapeutic agent and the second therapeutic
agent are administered either simultaneously or sequentially; and
further comprises assaying the expression of proteasomal activity
in a biological sample. In one embodiment the proteasomal activity
may be chymotrysin-like, trypsin-like, and/or caspase-like
activity. In one embodiment, the biological sample may be tumor,
peripheral blood mononuclear cells, or skin cells. The biological
sample may be obtained before, during, or after drug
administration. In one embodiment, the treatment method further
comprises adjusting the dose to achieve inhibition of about 20%,
about 25%, about 30%, about 35%, about 40%, about 45%, about 50%,
about 55%, about 60%, about 65%, about 70%, about 75%, about 80%,
about 85%, about 90%, about 95%, or about 100% of the proteasomal
activity. In one embodiment, the treatment method further comprises
adjusting the dose to achieve inhibition of at least 20%, at least
25%, at least 30%, at least 35%, at least 40%, at least 45%, at
least 50%, at least 55%, at least 60%, at least 65%, at least 70%,
at least 75%, at least 80%, at least 85%, at least 90%, or at least
95% of the proteasomal activity. In one embodiment, the treatment
method further comprises adjusting the dose to achieve inhibition
of about 20% to about 30%, about 30% to about 40%, about 40% to
about 50%, about 50% to about 60%, about 60% to about 70%, about
70% to about 80%, about 80% to about 90%, or greater than 90% of
the proteasomal activity.
[0233] In an aspect, provided herein are treatment methods, which
comprise administering to a subject in need of such treatment a
combination of a first therapeutic agent including an imipridone,
such as ONC201, 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:
[0234] (i) administering to the subject the first therapeutic
agent;
[0235] (ii) waiting until a predetermined waiting time has elapsed
after the time of administration of the first therapeutic agent to
the subject; and/or until adverse events are resolved or resolving;
and
[0236] (iii) administering the second therapeutic agent to the
subject, wherein the predetermined waiting time is chosen so as to
obtain a delayed therapeutic effect of the first therapeutic agent
without an increased risk of possible combined toxic effects of the
first and second therapeutic agents. In one embodiment, the
predetermined waiting time is determined based on the clearance
rate of the compound of the first therapeutic agent or a metabolite
thereof. In one embodiment, the predetermined waiting time is
determined by a quantitative assessment of renal function and
parameters of renal. In one embodiment, the predetermined waiting
time is determined by an assay for the determination of renal
function, wherein the assay is selected from the group consisting
of serum level the compound of the first therapeutic agent or a
metabolite thereof; clearance rate of the compound of the first
therapeutic agent or a metabolite thereof; 24-hour urinary
clearance of the compound of the first therapeutic agent or a
metabolite thereof.
[0237] In one embodiment of the treatment method, the predetermined
waiting time substantially equals the time required for systemic
clearance of the compound of the first therapeutic agent or a
metabolite thereof from the subject's body. In one embodiment of
the treatment method, the predetermined waiting time substantially
equals the time required for renal clearance of the compound of the
first therapeutic agent or a metabolite thereof from the subject's
body. In one embodiment of the treatment method, the predetermined
waiting time substantially equals the time required for hepatic
clearance of the compound of the first therapeutic agent or a
metabolite thereof from the subject's body. In one embodiment of
the treatment method, the predetermined waiting time substantially
equals the time required for total clearance of the compound of the
first therapeutic agent or a metabolite thereof from the subject's
body. In one embodiment of the treatment method, the predetermined
waiting time is about 4 hours. In other embodiments the waiting
time is 1 day. In one embodiment, the waiting time is until
C.sub.max of the compound of the first therapeutic agent has
passed. In other embodiments, the waiting time is after most of the
adverse events are resolved or are resolving. In one embodiment of
the treatment method, 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 treatment method, the
predetermined waiting time is a range of about 1-7 days, about 1-6
days, about 1-5 days, about 1-4 days, about 1-3 days, or about 1 to
2 days. In one embodiment, the waiting time is up to 3 weeks. The
preceeding are considered "therapeutic time periods."
[0238] When the order of administration is reversed, timing for the
administration of the first therapeutic agent can be after the
C.sub.max of the second therapeutic agent (i.e., the first
administered drug) has passed. In one embodiment, administration of
the first therapeutic agent can be after most or substantially all
of the first administered drug has been eliminated from the body or
the toxicity effects for the first administered drug are resolved
or are resolving.
[0239] In one embodiment, the treatment method further comprises
monitoring levels of the compound of the first therapeutic agent or
a metabolite thereof in the subject using pharmacokinetic
profiling. In some such embodiments, monitoring levels of the
compound of the first therapeutic agent or a metabolite thereof in
the subject using pharmacokinetic profiling comprises constructing
a pharmacokinetic profile of the compound of the first therapeutic
agent or a metabolite thereof for the subject using concentrations
of the compound of the first therapeutic agent or a metabolite
thereof in at least two samples obtained from the subject at time
points suitable to construct a pharmacokinetic profile. In one
embodiment, which include monitoring levels of the compound of the
first therapeutic agent or a metabolite thereof in the subject
using pharmacokinetic profiling, samples are collected from the
subject at point-of-care or point of use by sampling or
self-sampling on point-of-care devices or point of use devices or
on matrices suitable for storage of the samples prior to
quantitation in a laboratory. In one embodiment, each of the
point-of-care devices or point of use devices is capable of
quantitating the compound of the first therapeutic agent or a
metabolite thereof. In one embodiment, which include monitoring
levels of the compound of the first therapeutic agent or a
metabolite thereof in the subject, one or more samples are
collected from the subject at point-of-care or point of use by
biopsy device for analysis at the point-of-care or point of use
devices or for storage prior to analysis by a laboratory. In one
embodiment, a biopsy is taken after a time interval of 3-8 hours
following administration the first therapeutic agent to the
subject. In one embodiment, a biopsy is taken after a time interval
of 3-24 hours following administration of the first therapeutic
agent to the subject. In one embodiment, a biopsy is taken after a
time interval of 8-24 hours following administration of the first
therapeutic agent thereof to the subject. In one embodiment, a
biopsy is taken after a time interval of 2 days following
administration of the first therapeutic agent to the subject. In
one embodiment, a biopsy is taken after a time interval of 3 days
following administration of the first therapeutic agent to the
subject. In one embodiment, a biopsy is taken after a time interval
of 4 days following administration of the first therapeutic agent
to the subject. In one embodiment, a biopsy is taken after a time
interval of 1-7 days following administration of the first
therapeutic agent.
[0240] In one embodiment, the pharmacokinetic profile includes
pharmacokinetic parameters suitable for guiding dosing of the first
therapeutic agent for the subject being treated. In one embodiment
of the treatment method, the C.sub.max of the first therapeutic
agent following its administration to the subject ranges from about
1000 ng/dL to 1500 ng/dL for a therapeutic time period. In one
embodiment, C.sub.max is less than 1500 ng/dL and greater than 85
ng/dL for a therapeutic time period. In one embodiment, the
C.sub.max of the first therapeutic following its administration to
the subject ranges from about 1000 ng/mL to 1500 ng/mL for a
therapeutic time period. In one embodiment, C.sub.max is less than
1500 ng/mL and greater than 85 ng/mL for a therapeutic time
period.
[0241] In one embodiment, maximum concentration of the first
therapeutic agent in blood (whole blood, plasma, or serum)
("C.sub.max") of a subject after administering it to the subject is
a C.sub.max of from about 1000, 1010, 1020, 1030, 1040, 1050, 1060,
1070, 1080, 1090, 1100, 1110, 1120, 1130, 1140, 1150, 1160, 1170,
1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280,
1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390,
1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, or 1490 ng/dL
to about 1500 ng/dL; from about 100, 101, 102, 103, 104, 105, 106,
107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119,
120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132,
133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145,
146, 147, 148, or 149 ng/dL to about 150 ng/dL; or from about 10,
10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14, or 14.5 ng/dL to about 15
ng/dL.
[0242] In one embodiment, maximum concentration of the first
therapeutic agent in blood (whole blood, plasma, or serum) ("C.")
of the subject following its administration is a C. of 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 to about 1500 ng/mL;
from about 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110,
111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123,
124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136,
137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, or 149
ng/mL to about 150 ng/mL; or from about 10, 10.5, 11, 11.5, 120,
12.5, 13, 13.5, 14, or 14.5 ng/mL to about 15 ng/mL.
[0243] In one embodiment, maximum concentration of the first
therapeutic agent in blood (whole blood, plasma, or serum) ("C.")
of a subject following its administration is selected from about
1000, 1010, 1020, 1030, 1040, 1050, 1060, 1070, 1080, 1090, 1100,
1110, 1120, 1130, 1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210,
1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 1320,
1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430,
1440, 1450, 1460, 1470, 1480, or 1490 ng/dL. In one embodiment, the
C.sub.max of the first therapeutic agent in blood (whole blood,
plasma, or serum) ("C.sub.max") of a subject following its
administration is selected from about 100, 101, 102, 103, 104, 105,
106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,
119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131,
132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144,
145, 146, 147, 148, or 149 ng/dL. In one embodiment, the C.sub.max
of the first therapeutic agent following its administration is
selected from about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14, or
14.5 ng/dL.
[0244] In one embodiment, the C.sub.max of the first therapeutic
agent following its administration 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
C.sub.max of the first therapeutic agent following its
administration is selected from about 100, 101, 102, 103, 104, 105,
106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,
119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131,
132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144,
145, 146, 147, 148, or 149 ng/mL. In one embodiment, the C.sub.max
of the first therapeutic agent following its administration is
selected from about 10, 10.5, 11, 11.5, 120, 12.5, 13, 13.5, 14, or
14.5 ng/mL.
[0245] In one embodiment, the C.sub.max of the first therapeutic
agent following its administration 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 one embodiment, the
C.sub.max of the first therapeutic agent following its
administration is selected 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 149 ng/dL. In
one embodiment, the C.sub.max of the first therapeutic agent
following its administration is selected from about 1, 1.5, 2, 2.5,
3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5,
11, 11.5, 12, 12.5, 13, 13.5, 14, or 14.5 ng/dL.
[0246] In one embodiment, the C.sub.max of the first therapeutic
agent following its administration 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/mL. In one embodiment, the
C.sub.max of the first therapeutic following its administration is
selected 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 149 ng/mL. In one embodiment, the
C.sub.max of the first therapeutic agent following its
administration is selected from about 1, 1.5, 2, 2.5, 3, 3.5, 4,
4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5,
12, 12.5, 13, 13.5, 14, or 14.5 ng/mL.
[0247] In one embodiment, the C.sub.max of the first therapeutic
agent after administering it to the subject ranges from about 85
ng/dL to 1500 ng/dL; from about 8.5 ng/dL to 150 ng/dL; or from
about 0.85 ng/dL to 15 ng/dL. In one embodiment, the C.sub.max of
the first therapeutic agent in a subject's blood (whole blood,
plasma, or serum) after its administration 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, or 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 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 14.5
ng/dL to about 15 ng/dL.
[0248] In one embodiment, the C.sub.max of the first therapeutic
agent following its administration ranges from about 85 ng/mL to
1500 ng/mL; from about 8.5 ng/mL to 150 ng/mL; or from about 0.85
ng/mL to 15 ng/mL. In one embodiment, the C.sub.max of the first
therapeutic following its administration 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, or 1495 ng/mL to about 1500 ng/mL;
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 149 ng/mL to about 150 ng/mL; 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 14.5
ng/mL to about 15 ng/mL.
[0249] In one embodiment, the total drug exposure over time,
measured as the area under the curve ("AUC") of a plot of the
concentration of the drug in blood (whole blood, plasma, or serum)
of a subject following administration of the drug against time
after administration of the drug ranges from about 150 ng hr/mL to
about 8000 ng hr/mL; from about 15 ng hr/mL to about 800 ng hr/mL;
or from about 1.5 ng hr/mL to about 80 ng hr/mL. In one embodiment,
AUC is less than 8000 ng hr/mL and is greater than or equal to 150
ng hr/mL. In one embodiment, AUC is less than 800 ng hr/mL and is
greater than or equal to 15 ng hr/mL. In one embodiment, AUC is
less than 80 ng hr/mL and is greater than or equal to 1.5 ng
hr/mL.
[0250] In one embodiment, the total drug exposure over time is an
AUC of from about 100 ng hr/mL to about 8000 ng hr/mL; from about
10 ng hr/mL to about 800 ng hr/mL; or from about 1 ng hr/mL to
about 80 ng hr/mL. In one embodiment, the total drug exposure over
time is an AUC of from 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 7800 ng hr/mL to about 8000 ng hr/mL. In one embodiment,
the total drug exposure over time is an AUC of from about 15, 20,
40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300,
320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560,
580, 600, 620, 640, 660, 680, 700, 720, 740, 760, or 780 ng hr/mL
to about 800 ng hr/mL. In one embodiment, the total drug exposure
over time is an AUC of from about from about 1.5, 2, 4, 6, 8, 10,
12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44,
46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, or
78 ng hr/mL to about 80 ng hr/mL.
[0251] In one embodiment, the total drug exposure over time is an
AUC of from about 100 ng hr/mL to about 8000 ng hr/mL, from about
10 ng hr/mL to about 800 ng hr/mL; or from about 1 ng hr/mL to
about 80 ng hr/mL. In one embodiment, the total drug exposure over
time is an AUC of from about from about 150 ng hr/mL to about 7800,
7600, 7400, 7200, 7000, 6800, 6600, 6400, 6200, 6000, 5800, 5600,
5400, 5200, 5000, 4800, 4600, 4400, 4200, 4000, 3800, 3600, 3400,
3200, 3000, 2800, 2600, 2400, 2200, 2000, 1800, 1600, 1400, 1200,
1000, 800, 600, 400, or 200 ng hr/mL. In one embodiment, the total
drug exposure over time is an AUC of from about from about 15 ng
hr/mL to about 780, 760, 740, 720, 700, 680, 660, 640, 620, 600,
580, 560, 540, 520, 500, 480, 460, 440, 420, 400, 380, 360, 340,
320, 300, 280, 260, 240, 220, 200, 180, 160, 140, 120, 100, 80, 60,
40, or 20 ng hr/mL. In one embodiment, the total drug exposure over
time is an AUC of from about from about 1.5 ng hr/mL to about 78,
76, 74, 72, 70, 68, 66, 64, 62, 60, 58, 56, 54, 52, 50, 48, 46, 44,
42, 40, 38, 36, 34, 32, 30, 28, 26, 24, 22, 20, 18, 16, 14, 12, 10,
8, 6, 4, or 2 ng hr/mL. In one embodiment, the total drug exposure
over time is an AUC of from about 100 ng hr/mL to about 200 ng
hr/mL; from about 10 ng hr/mL to about 20 ng hr/mL; or from about 1
ng hr/mL to about 2 ng hr/mL.
[0252] In one embodiment, the total drug exposure over time is an
AUC selected from about 100, 150, 200, 400, 600, 800, 1000, 1200,
1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400,
3600, 3800, 4000, 4200, 4400, 46000, 4800, 5000, 5200, 5400, 5600,
5800, 6000, 6200, 6400, 6600, 6800, 7000, 7200, 7400, 7600, 7800,
and 8000 ng hr/mL. In one embodiment, the total drug exposure over
time is an AUC selected from about 10, 15, 20, 40, 60, 80, 100,
120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360,
380, 400, 420, 440, 4600, 480, 500, 520, 540, 560, 580, 600, 620,
640, 660, 680, 700, 720, 740, 760, 780, and 800 ng hr/mL. In one
embodiment, the total drug exposure over time is an AUC selected
from about 1, 15, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26,
28, 30, 32, 34, 36, 38, 40, 42, 44, 460, 48, 50, 52, 54, 56, 58,
60, 62, 64, 66, 68, 70, 72, 74, 76, 78, and 80 ng hr/mL.
[0253] In another aspect, provided herein are methods of treatment,
or use of a composition to treat a disease state, which comprises
administering to a subject in need of such treatment a combination
of a first therapeutic agent and a second therapeutic agent, the
method comprising:
[0254] (i) administering to the subject the first therapeutic agent
including an imipridone, such as ONC201, an analog thereof, or a
pharmaceutically acceptable salt thereof;
[0255] (ii) monitoring levels of the compound of the first
therapeutic agent or a metabolite thereof in the subject using
pharmacokinetic profiling; and
[0256] (iii) administering the second therapeutic agent conditional
on the level of the first therapeutic agent in the subject. In one
embodiment, the monitoring step includes constructing a
pharmacokinetic profile of the compound of the first therapeutic
agent or a metabolite thereof for the subject using concentrations
of the compound of the first therapeutic agent or a metabolite
thereof in a plurality of samples obtained from the subject at time
points suitable to construct a pharmacokinetic profile. In one
embodiment, at least two samples are collected at point-of-care or
point of use by sampling or self-sampling on point-of-care devices
or point of use devices or on matrices suitable for storage of the
samples prior to quantitation of the compound or a metabolite
thereof by a laboratory. In one embodiment, each point-of-care
devices or point of use devices is capable of quantitating the
compound or a metabolite thereof. In one embodiment, the
pharmacokinetic profile includes pharmacokinetic parameters
suitable for guiding dosing of the compound or a salt thereof for
the subject. In one embodiment, the samples include from 2-12
samples. In one embodiment, the samples are collected over a time
period of up to 8 hours, up to 24 hours, up to 48 hours, or up to
72 hours. In one embodiment, the pharmacokinetic parameters include
at least one parameter selected from the group consisting of AUC,
AUC.sub.inf, T.sub.max, C.sub.max, time above threshold, steady
state concentration, absorption rate, clearance rate, distribution
rate, terminal T-1/2 or parameters drawn from noncompartmental
pharmacokinetic (PK) or compartmental PK analysis, including
physiological model-based compartmental PK analysis. In one
embodiment, the treatment method further comprises generating a
report including the pharmacokinetic profile of the subject. In one
embodiment, the report includes a recommendation regarding dosing
based on the pharmacokinetic profile of the subject. In one
embodiment, a reduction in dosage of ONC201, the analog thereof, or
the pharmaceutically acceptable salt thereof is indicated to reduce
risk of toxicity based on one or more pharmacokinetic parameters.
In one embodiment, the reduction in dosage of the compound or salt
thereof is indicated based on time above threshold, wherein the
threshold is the drug concentration above which toxicity occurs, or
one or more of AUC, AUC.sub.inf, mean residence time (MRT),
exponentials defining the pharmacokinetic profile, volume of
distribution at steady state (Vss), volume of distribution during
the terminal phase (Vz) or combination of a group of
pharmacokinetic variable to adequately describe the pharmacokinetic
profile. In one embodiment, a dose adjustment of the compound or
salt thereof is indicated to increase efficacy based on one or more
pharmacokinetic parameters. In one embodiment, an increase in
dosage of the compound or salt thereof is indicated based on one or
more of AUC, AUC.sub.inf, MRT, exponentials defining the
pharmacokinetic profile, steady state volume (Vss) of distribution,
volume of distribution during the terminal phase (Vz) or
combination of a group of pharmacokinetic variables to adequately
describe the pharmacokinetic profile. In one embodiment, the dose
of the compound or salt thereof is adjusted to within 5% to 25% of
a desired target value. In one embodiment, each of the samples is
applied to the point-of-care device or the point of use device for
determining the concentration of the compound or a metabolite
thereof, wherein the point-of-care device or the point of use
device comprises a lateral flow strip having a construction and
composition such that an application of one or more of the samples
to the lateral flow strip causes a fraction of the drug in the
sample to bind to with a component of the lateral flow strip such
that a detectable signal proportional to the concentration of the
drug in the applied sample is produced. In one embodiment, the
samples are applied to matrices suitable for storage of the samples
prior to quantitation by a laboratory. In one embodiment, the
samples are stored as dried blood spots. In one embodiment, drug
concentrations are measured by ELISA, LC MS MS, LC UV or LCMS. In
one embodiment, the pharmacokinetic parameters include at least one
of steady state concentration, absorption, and terminal T.sub.1/2.
In one embodiment, at least one of the samples is whole blood.
IX. MULTIMODAL THERAPEUTIC METHODS
[0257] In one aspect, provided herein are multimodal therapeutic
methods in which administration of an imipridone, such as ONC201,
an analog thereof, or a pharmaceutically acceptable salt thereof to
a subject in need of such treatment is supplemented by
administration of other therapeutic modalities. In one embodiment,
the multimodal therapeutic method comprises administering to a
subject a pharmaceutical composition comprising an imipridone, such
as ONC201, an analog thereof, or a pharmaceutically acceptable salt
thereof in conjunction with radiation therapy or after radiation is
determined to not have been efficacious. In one embodiment, the
multimodal therapeutic method comprises administering to a subject
a pharmaceutical composition comprising an imipridone, such as
ONC201, an analog thereof, or a pharmaceutically acceptable salt
thereof in conjunction with radiation therapy, wherein the
pharmaceutical composition comprising theimipridone, such as
ONC201, the analog thereof, or pharmaceutically acceptable salt
thereof and the radiation therapy are administered concurrently or
sequentially in any order. In one embodiment, the multimodal
therapeutic method comprises administering to a subject a
pharmaceutical composition comprising an imipridone, such as
ONC201, an analog thereof, or a pharmaceutically acceptable salt
thereof in conjunction with radiation therapy in a sequential
arrangement. In one embodiment, the multimodal therapeutic method
comprises administering to a subject in need of such treatment a
pharmaceutical composition comprising an imipridone, such as
ONC201, an analog thereof, or a pharmaceutically acceptable salt
thereof concurrently with radiation therapy. In one embodiment, the
multimodal therapeutic method is used for the treatment of cancer.
In one embodiment, the multimodal therapeutic method includes
administering to a cancer subject in need of such treatment a
pharmaceutical composition comprising an imipridone, such as
ONC201, an analog thereof, or a pharmaceutically acceptable salt
thereof and irradiating cancer cells with a radiation beam. In one
embodiment, the multimodal therapeutic method uses the technique of
conformal radiotherapy (CRT) to deliver a dose volume histogram
(DVH) prescribed to a cancer subject. In one embodiment, the
multimodal therapeutic method uses the technique of intensity
modulated radiation therapy (IMRT) to deliver radiation to cancer
cells. In one embodiment, the multimodal therapeutic method uses
techniques that compensate for motion of tumors in the subject
during treatment (e.g., where doses of radiation must be
administered to a thoracic tumor which moves as the patient
breathes). For example, the multimodal therapeutic method use Four
Dimensional Computed Tomography (4D CT) scanning techniques to
adjust the delivered radiation field to compensate for tumor motion
over the breathing cycle.
[0258] Any suitable type of radiation, including gamma radiation
which is given fractionated, IMRT (intensity modulated radiation
therapy), gamma knife, proton therapy and brachytherapy can be used
with the multimodal therapeutic method. Radiation therapy and
administering an imipridone, such as ONC201, an analog thereof, or
a pharmaceutically acceptable salt thereof can be used to treat
brain tumors such as glioblastoma or disease that has metastasized
to the brain from lung cancer. The multimodal therapeutic method
can be used to treat lung cancer, pancreatic cancer, rectal cancer,
breast cancer, sarcoma, prostate cancer, gynecological
malignancies, and lymphoma. The gamma knife is used frequently to
treat brain metastases. In one embodiment, the multimodal
therapeutic method includes use of proton therapy to treat cancer,
including brain tumors, prostate cancer and any tumor proximate
vital organs where it is very important to minimize toxicity to
nearby normal tissue.
[0259] In one embodiment, a multimodal therapeutic method includes
administering to a cancer subject in need of such treatment a
pharmaceutical composition comprising an imipridone, such as
ONC201, an analog thereof, or a pharmaceutically acceptable salt
thereof in combination with adoptive cell therapy (e.g., CAR-T
(JCAR 14, 15, 16, 17, KTE-C19, or CTL019); other T Cell (AFM13); or
NK (CDNO-109 or NK-92)) either simultaneously or in
combination.
[0260] In one embodiment, the multimodal therapeutic method
eliminates minimal residual disease without adding to toxicity
resulting from treatment by an imipridone, such as ONC201, an
analog thereof, or a pharmaceutically acceptable salt thereof. In
one embodiment, the multimodal therapeutic method improves
prognosis and/or reduces adverse side-effects associated with a
disease state or condition in a subject undergoing treatment.
X. ADDITIONAL IMIPRIDONE DERIVATIVES, ANALOGS, AND SALTS
[0261] In one aspect, provided herein are compounds that are
analogs of the compounds of formula (10) and methods of making
them. Persons skilled in the art will understand that the general
principles and concepts described above in conjunction with ONC201
and compounds of formula (10) and their salts, including principles
and concepts related to methods and pharmaceutical compositions,
apply with equal force to the following analogs and salts
thereof.
[0262] In one embodiment, the analogs have the structure of
compound (25):
##STR00018##
wherein Y is NR.sub.4 or O, and wherein R.sub.1, R.sub.2, R.sub.3,
and R.sub.4 independently represent H, alkyl, cycloalkyl,
cycloalkylalkyl, carboxyl, haloalkyl, alkenyl, cycloalkenyl,
alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl,
alkoxycarbonyl, aralkoxy, aralkylthio, alkanoyl, mercapto,
alkylthio, arylthio, alkylsulfinyl, arylsulfinyl, alkylsulfonyl,
arylsulfonyl, heteroaryl, acyl, and heterocycle radicals. In one
embodiment, R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are optionally
substituted. In one embodiment, some or all hydrogens in R.sub.1,
R.sub.2, R.sub.3, and R.sub.4 are substituted by deuterium. In
other embodiments, R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are
independently selected from the group consisting of H,
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
C.sub.1-4 benzyl-piperazine, and C.sub.1-4alkylthienyl, wherein
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
and C.sub.1-4benzyl-piperazine are optionally substituted with
C.sub.1-4alkyl, hydroxyl, or halo. In still other embodiments,
R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are independently selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph.
[0263] In one embodiment, the analogs have the structure of
compound (26):
##STR00019##
wherein R.sub.1 and R.sub.2 independently represent H, alkyl,
cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy,
aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio,
alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl,
arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and
heterocycle radicals. In one embodiment, R.sub.1 and R.sub.2 are
independently selected from the group consisting of H,
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
C.sub.1-4benzyl-piperazine, and C.sub.1-4alkylthienyl, wherein
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
and C.sub.1-4benzyl-piperazine are optionally substituted with
C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl, perhalogenated
C.sub.1-4alkyl, or halo. In one embodiment, R.sub.1 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph. In one embodiment, R.sub.2 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph.
[0264] In one embodiment, R.sub.1 is a benzyl optionally
substituted with one or more of the following substituents alone or
in combination in the ortho, meta, and/or para positions of the
benzyl ring: --CH.sub.3, --NO.sub.2, --OCH.sub.3, --CXH.sub.2,
--CX.sub.2H, --CX.sub.3, --CH.sub.2(CX.sub.3),
--CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1,
--OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p is an integer from 2
to 20 and where X is a halogen including F, Cl, Br, or I;
preferably, F, Cl, or Br; more preferably, F or Cl. In one
embodiment, R.sub.2 is a benzyl substituted with one or more of the
following substituents alone or in combination in the ortho, meta,
and/or para positions of the benzyl ring: --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CXH.sub.2, --CX.sub.2H, --CX.sub.3,
--CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3,
--C.sub.pX.sub.2p+1, --OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p
is an integer from 2 to 20 and where X is a halogen.
[0265] In one embodiment, R.sub.1 is a H. In one embodiment,
R.sub.1 is a substituted or an unsubstituted arylalkyl, such as a
benzyl or phenylethyl group. In one embodiment, the arylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo.
[0266] In one embodiment, R.sub.2 is a substituted or an
unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In
one embodiment, the arylalkyl is substituted with C.sub.1-4alkyl,
C.sub.1-4alkoxyl, hydroxyl, perhalogenated C.sub.1-4alkyl, or halo.
In one embodiment, the arylalkyl is substituted with one or more
substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3. In one embodiment, R.sub.2
is a substituted or an unsubstituted heterocycloalkylalkyl, such as
a morpholinoalkyl or piperazinylalkyl group. In one embodiment,
R.sub.2 is a substituted or an unsubstituted heteroarylalkyl, such
as an isoxazolidinylmethyl or pyridylmethyl group. In one
embodiment, the heterocycloalkylalkyl or heteroarylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo. In one embodiment, the
heterocycloalkylalkyl or heteroarylalkyl is substituted with one or
more substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3.
[0267] In one embodiment, the analogs have the structure of
compound (27):
##STR00020##
wherein R.sub.1 is H, alkyl, cycloalkyl, cycloalkylalkyl, carboxyl,
haloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, aralkyl,
hydroxyalkyl, alkoxy, aryloxy, alkoxyalkyl, alkoxycarbonyl,
aralkoxy, aralkylthio, alkanoyl, mercapto, alkylthio, arylthio,
alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl,
heteroaryl, acyl, and heterocycle radicals. In one embodiment,
R.sub.1 is selected from the group consisting of H, C.sub.1-4alkyl,
C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone, C.sub.1-4
benzyl-piperazine, and C.sub.1-4alkylthienyl, wherein
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
and C.sub.1-4benzyl-piperazine are optionally substituted with
C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl, perhalogenated
C.sub.1-4alkyl, or halo. In one embodiment, R.sub.1 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph.
[0268] In one embodiment, R.sub.1 is a benzyl optionally
substituted with one or more of the following substituents alone or
in combination in the ortho, meta, and/or para positions of the
benzyl ring: --CH.sub.3, --NO.sub.2, --OCH.sub.3, --CXH.sub.2,
--CX.sub.2H, --CX.sub.3, --CH.sub.2(CX.sub.3),
--CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1,
--OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p is an integer from 2
to 20 and where X is a halogen including F, Cl, Br, or I;
preferably, F, Cl, or Br; more preferably, F or Cl. In one
embodiment, R.sub.1 is a H. In one embodiment, R.sub.1 is a
substituted or an unsubstituted arylalkyl, such as a benzyl or
phenylethyl group. In one embodiment, the arylalkyl is substituted
with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl, perhalogenated
C.sub.1-4alkyl, or halo.
[0269] In one embodiment, the analogs have the structure of
compound (28):
##STR00021##
wherein R.sub.1 and R.sub.2 independently represent H, alkyl,
cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy,
aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio,
alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl,
arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and
heterocycle radicals. In one embodiment, R.sub.1 and R.sub.2 are
independently selected from the group consisting of H,
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
C.sub.1-4 benzyl-piperazine, and C.sub.1-4alkylthienyl, wherein
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
and C.sub.1-4benzyl-piperazine are optionally substituted with
C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl, perhalogenated
C.sub.1-4alkyl, or halo. In one embodiment, R.sub.1 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2-(2,4-di F-Ph), CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), and
(CH.sub.2).sub.3CO-4F-Ph. In one embodiment, R.sub.2 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph. In one embodiment, when R.sub.1 is
CH.sub.2Ph, R.sub.2 is not CH.sub.2-(2-CH.sub.3-Ph). In one
embodiment, R.sub.1 is CH.sub.2Ph and R.sub.2 is
CH.sub.2-(2-CH.sub.3-Ph). In one embodiment, R.sub.1 is CH.sub.2Ph
and R.sub.2 is CH.sub.2-(2,4-di F-Ph). In one embodiment, R.sub.1
is CH.sub.2Ph and R.sub.2 is CH.sub.2-(4-CF.sub.3-Ph).
[0270] In one embodiment, R.sub.1 is a benzyl optionally
substituted with one or more of the following substituents alone or
in combination in the ortho, meta, and/or para positions of the
benzyl ring: --CH.sub.3, --NO.sub.2, --OCH.sub.3, --CXH.sub.2,
--CX.sub.2H, --CX.sub.3, --CH.sub.2(CX.sub.3),
--CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1,
--OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p is an integer from 2
to 20 and where X is a halogen including F, Cl, Br, or I;
preferably, F, Cl, or Br; more preferably, F or Cl. In one
embodiment, R.sub.2 is a benzyl substituted with one or more of the
following substituents alone or in combination in the ortho, meta,
and/or para positions of the benzyl ring: --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CXH.sub.2, --CX.sub.2H, --CX.sub.3,
--CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3,
--C.sub.pX.sub.2p+1, --OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p
is an integer from 2 to 20 and where X is a halogen.
[0271] In one embodiment, R.sub.1 is a H. In one embodiment,
R.sub.1 is a substituted or an unsubstituted arylalkyl, such as a
benzyl or phenylethyl group. In one embodiment, the arylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo.
[0272] In one embodiment, R.sub.2 is a substituted or an
unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In
one embodiment, the arylalkyl is substituted with C.sub.1-4alkyl,
C.sub.1-4alkoxyl, hydroxyl, perhalogenated C.sub.1-4alkyl, or halo.
In one embodiment, the arylalkyl is substituted with one or more
substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3. In one embodiment, R.sub.2
is a substituted or an unsubstituted heterocycloalkylalkyl, such as
a morpholinoalkyl or piperazinylalkyl group. In one embodiment,
R.sub.2 is a substituted or an unsubstituted heteroarylalkyl, such
as an isoxazolidinylmethyl or pyridylmethyl group. In one
embodiment, the heterocycloalkylalkyl or heteroarylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo. In one embodiment, the
heterocycloalkylalkyl or heteroarylalkyl is substituted with one or
more substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3.
[0273] In one embodiment, the analogs have the structure of
compound (29):
##STR00022##
wherein R.sub.1 and R.sub.2 independently represent H, alkyl,
cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy,
aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio,
alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl,
arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and
heterocycle radicals. In one embodiment, R.sub.1 and R.sub.2 are
independently selected from the group consisting of H,
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
C.sub.1-4 benzyl-piperazine, and C.sub.1-4alkylthienyl, wherein
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
and C.sub.1-4benzyl-piperazine are optionally substituted with
C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl, perhalogenated
C.sub.1-4alkyl, or halo. In one embodiment, R.sub.1 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph. In one embodiment, R.sub.2 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph. In one embodiment, when R.sub.1 is
CH.sub.2Ph, R.sub.2 is not CH.sub.2-(2-CH.sub.3-Ph). In one
embodiment, R.sub.1 is CH.sub.2Ph and R.sub.2 is
CH.sub.2-(2-CH.sub.3-Ph). In one embodiment, R.sub.1 is CH.sub.2Ph
and R.sub.2 is CH.sub.2-(2,4-di F-Ph). In one embodiment, R.sub.1
is CH.sub.2Ph and R.sub.2 is CH.sub.2-(4-CF.sub.3-Ph).
[0274] In one embodiment, R.sub.1 is a benzyl optionally
substituted with one or more of the following substituents alone or
in combination in the ortho, meta, and/or para positions of the
benzyl ring: --CH.sub.3, --NO.sub.2, --OCH.sub.3, --CXH.sub.2,
--CX.sub.2H, --CX.sub.3, --CH.sub.2(CX.sub.3),
--CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1,
--OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p is an integer from 2
to 20 and where X is a halogen including refers to F, Cl, Br, or I;
preferably, F, Cl, or Br; more preferably, F or Cl. In one
embodiment, R.sub.2 is a benzyl substituted with one or more of the
following substituents alone or in combination in the ortho, meta,
and/or para positions of the benzyl ring: --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CXH.sub.2, --CX.sub.2H, --CX.sub.3,
--CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3,
--C.sub.pX.sub.2p+1, --OCX3, or --OC.sub.pX.sub.2p+1, where p is an
integer from 2 to 20 and where X is a halogen.
[0275] In one embodiment, R.sub.1 is a H. In one embodiment,
R.sub.1 is a substituted or an unsubstituted arylalkyl, such as a
benzyl or phenylethyl group. In one embodiment, the arylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo.
[0276] In one embodiment, R.sub.2 is a substituted or an
unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In
one embodiment, the arylalkyl is substituted with C.sub.1-4alkyl,
C.sub.1-4alkoxyl, hydroxyl, perhalogenated C.sub.1-4alkyl, or halo.
In one embodiment, the arylalkyl is substituted with one or more
substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3. In one embodiment, R.sub.2
is a substituted or an unsubstituted heterocycloalkylalkyl, such as
a morpholinoalkyl or piperazinylalkyl group. In one embodiment,
R.sub.2 is a substituted or an unsubstituted heteroarylalkyl, such
as an isoxazolidinylmethyl or pyridylmethyl group. In one
embodiment, the heterocycloalkylalkyl or heteroarylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo. In one embodiment, the
heterocycloalkylalkyl or heteroarylalkyl is substituted with one or
more substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3.
[0277] In one embodiment, the analogs have the structure of
compound (30):
##STR00023##
wherein R.sub.1 and R.sub.2 independently represent H, alkyl,
cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy,
aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio,
alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl,
arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and
heterocycle radicals. In one embodiment, R.sub.1 and R.sub.2 are
independently selected from the group consisting of H,
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
C.sub.1-4 benzyl-piperazine, and C.sub.1-4alkylthienyl, wherein
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
and C.sub.1-4benzyl-piperazine are optionally substituted with
C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl, perhalogenated
C.sub.1-4alkyl, or halo. In one embodiment, R.sub.1 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph. In one embodiment, R.sub.2 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph. In one embodiment, when R.sub.1 is
CH.sub.2Ph, R.sub.2 is not CH.sub.2-(2-CH.sub.3-Ph). In one
embodiment, R.sub.1 is CH.sub.2Ph and R.sub.2 is
CH.sub.2-(2-CH.sub.3-Ph). In one embodiment, R.sub.1 is CH.sub.2Ph
and R.sub.2 is CH.sub.2-(2,4-di F-Ph). In one embodiment, R.sub.1
is CH.sub.2Ph and R.sub.2 is CH.sub.2-(4-CF.sub.3-Ph).
[0278] In one embodiment, R.sub.1 is a benzyl optionally
substituted with one or more of the following substituents alone or
in combination in the ortho, meta, and/or para positions of the
benzyl ring: --CH.sub.3, --NO.sub.2, --OCH.sub.3, --CXH.sub.2,
--CX.sub.2H, --CX.sub.3, --CH.sub.2(CX.sub.3),
--CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1,
--OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p is an integer from 2
to 20 and where X is a halogen including refers to F, Cl, Br, or I,
preferably, F, Cl, or Br, more preferably, F or Cl. In one
embodiment, R.sub.2 is a benzyl substituted with one or more of the
following substituents alone or in combination in the ortho, meta,
and/or para positions of the benzyl ring: --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CXH.sub.2, --CX.sub.2H, --CX.sub.3,
--CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3,
--C.sub.pX.sub.2p+1, --OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p
is an integer from 2 to 20 and where X is a halogen.
[0279] In one embodiment, R.sub.1 is a H. In one embodiment,
R.sub.1 is a substituted or an unsubstituted arylalkyl, such as a
benzyl or phenylethyl group. In one embodiment, the arylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo.
[0280] In one embodiment, R.sub.2 is a substituted or an
unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In
one embodiment, the arylalkyl is substituted with C.sub.1-4alkyl,
C.sub.1-4alkoxyl, hydroxyl, perhalogenated C.sub.1-4alkyl, or halo.
In one embodiment, the arylalkyl is substituted with one or more
substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3. In one embodiment, R.sub.2
is a substituted or an unsubstituted heterocycloalkylalkyl, such as
a morpholinoalkyl or piperazinylalkyl group. In one embodiment,
R.sub.2 is a substituted or unsubstituted heteroarylalkyl, such as
an isoxazolidinylmethyl or pyridylmethyl group. In one embodiment,
the heterocycloalkylalkyl or heteroarylalkyl is substituted with
C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl, perhalogenated
C.sub.1-4alkyl, or halo. In one embodiment, the
heterocycloalkylalkyl or heteroarylalkyl is substituted with one or
more substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3.
[0281] In one embodiment, the analogs have the structure of
compound (31):
##STR00024##
wherein R.sub.1 and R.sub.2 independently represent H, alkyl,
cycloalkyl, cycloalkylalkyl, carboxyl, haloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, aralkyl, hydroxyalkyl, alkoxy,
aryloxy, alkoxyalkyl, alkoxycarbonyl, aralkoxy, aralkylthio,
alkanoyl, mercapto, alkylthio, arylthio, alkylsulfinyl,
arylsulfinyl, alkylsulfonyl, arylsulfonyl, heteroaryl, acyl, and
heterocycle radicals. In one embodiment, R.sub.1 and R.sub.2 are
independently selected from the group consisting of H,
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
C.sub.1-4benzyl-piperazine, and C.sub.1-4alkylthienyl, wherein
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
and C.sub.1-4benzyl-piperazine are optionally substituted with
C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl, perhalogenated
C.sub.1-4alkyl, or halo. In one embodiment, R.sub.1 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph. In one embodiment, R.sub.2 is selected
from the group consisting of H, CH.sub.3, CH.sub.2Ph,
CH.sub.2-((2-Cl)-Ph), CH.sub.2-(2-thienyl), CH.sub.2CH.sub.2Ph,
CH.sub.2CH.sub.2(4-N-benzyl-piperazine), CH.sub.2-(2,4-di F-Ph),
CH.sub.2-((2-CH.sub.3)-Ph), CH.sub.2CHOHPh, and
(CH.sub.2).sub.3CO-4F-Ph. In one embodiment, when R.sub.1 is
CH.sub.2Ph, R.sub.2 is not CH.sub.2-(2-CH.sub.3-Ph). In one
embodiment, R.sub.1 is CH.sub.2Ph and R.sub.2 is
CH.sub.2-(2-CH.sub.3-Ph). In one embodiment, R.sub.1 is CH.sub.2Ph
and R.sub.2 is CH.sub.2-(2,4-di F-Ph). In one embodiment, R.sub.1
is CH.sub.2Ph and R.sub.2 is CH.sub.2-(4-CF.sub.3-Ph).
[0282] In one embodiment, R.sub.1 is a benzyl optionally
substituted with one or more of the following substituents alone or
in combination in the ortho, meta, and/or para positions of the
benzyl ring: --CH.sub.3, --NO.sub.2, --OCH.sub.3, --CXH.sub.2,
--CX.sub.2H, --CX.sub.3, --CH.sub.2(CX.sub.3),
--CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1,
--OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p is an integer from 2
to 20 and where X is a halogen including F, Cl, Br, or I;
preferably, F, Cl, or Br; more preferably, F or Cl. In one
embodiment, R.sub.2 is a benzyl substituted with one or more of the
following substituents alone or in combination in the ortho, meta,
and/or para positions of the benzyl ring: --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CXH.sub.2, --CX.sub.2H, --CX.sub.3,
--CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2, --C(CX.sub.3).sub.3,
--C.sub.pX.sub.2p+1, --OCX.sub.3, or --OC.sub.pX.sub.2p+1, where p
is an integer from 2 to 20 and where X is a halogen.
[0283] In one embodiment, R.sub.1 is a H. In one embodiment,
R.sub.1 is a substituted or an unsubstituted arylalkyl, such as a
benzyl or phenylethyl group. In one embodiment, the arylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo.
[0284] In one embodiment, R.sub.2 is a substituted or an
unsubstituted arylalkyl, such as a benzyl or phenylethyl group. In
one embodiment, the arylalkyl is substituted with C.sub.1-4alkyl,
C.sub.1-4alkoxyl, hydroxyl, perhalogenated C.sub.1-4alkyl, or halo.
In one embodiment, the arylalkyl is substituted with one or more
substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3. In one embodiment, R.sub.2
is a substituted or an unsubstituted heterocycloalkylalkyl, such as
a morpholinoalkyl or piperazinylalkyl group. In one embodiment,
R.sub.2 is a substituted or an unsubstituted heteroarylalkyl, such
as an isoxazolidinylmethyl or pyridylmethyl group. In one
embodiment, the heterocycloalkylalkyl or heteroarylalkyl is
substituted with C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl,
perhalogenated C.sub.1-4alkyl, or halo. In one embodiment, the
heterocycloalkylalkyl or heteroarylalkyl is substituted with one or
more substituents selected from the group consisting of halo,
--CH.sub.3, --CF.sub.3, and --OCH.sub.3.
[0285] In one embodiment, provided herein are compounds of formula
(100):
##STR00025##
wherein R.sub.1 and R.sub.2 are independently selected from H,
alkyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl,
heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl,
heteroarylalkyl, alkoxyalkyl, alkoxycarbonyl, aralkoxy,
aralkylthio, and acyl radicals. In one embodiment, R.sub.1 is
CH.sub.2Ph and R.sub.2 is CH.sub.2-(2-CH.sub.3-Ph), which is an
ONC201 linear isomer (i.e., TIC-10)
##STR00026##
which lacks anti-cancer activity (Jacob et al., Angew. Chem. Int.
Ed., (2014) 53:6628; Wagner et al., Oncotarget (2015) 5(24):12728).
But as shown in the Examples TIC-10 is a CXCR7 agonist. CXCR7
agonists can be used for liver regeneration and preventing or
treating liver fibrosis (Nature (2014) 505:97).
[0286] In one embodiment, R.sub.1 and R.sub.2 are independently
selected from the group consisting of H, C.sub.1-4alkyl,
C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone, C.sub.1-4
benzyl-piperazine, C.sub.1-4alkylthienyl, C.sub.1-4alkylpyridinyl,
C.sub.1-4alkylisoxazolidinyl, C.sub.1-4alkylmorpholinyl,
C.sub.1-4alkylthiazolyl, and C.sub.1-4alkylpyrazinyl wherein
C.sub.1-4alkyl, C.sub.1-4alkylphenyl, C.sub.1-4alkylphenylketone,
C.sub.1-4 benzyl-piperazine, C.sub.1-4alkylthienyl,
C.sub.1-4alkylpyridinyl, C.sub.1-4alkylisoxazolidinyl,
C.sub.1-4alkylmorpholinyl, C.sub.1-4alkylthiazolyl, and
C.sub.1-4alkylpyrazinyl are optionally substituted with
C.sub.1-4alkyl, C.sub.1-4alkoxyl, hydroxyl, perhalogenated
C.sub.1-4alkyl, or halo. In one embodiment, R.sub.1 and/or R.sub.2
is a substituted or unsubstituted, arylalkyl or heteroarylalkyl. In
one embodiment, the heteroarylalkyl is selected from
C.sub.1-4alkylpyrrolyl, C.sub.1-4alkylfuryl, C.sub.1-4alkylpyridyl,
C.sub.1-4alkyl-1,2,4-thiadiazolyl, C.sub.1-4alkylpyrimidyl,
C.sub.1-4alkylthienyl, C.sub.1-4alkylisothiazolyl,
C.sub.1-4alkylimidazolyl, C.sub.1-4alkyltetrazolyl,
C.sub.1-4alkylpyrazinyl, C.sub.1-4alkylpyrimidyl,
C.sub.1-4alkylquinolyl, C.sub.1-4alkylisoquinolyl,
C.sub.1-4alkylthiophenyl, C.sub.1-4alkylbenzothienyl,
C.sub.1-4alkylisobenzofuryl, C.sub.1-4alkylpyrazolyl,
C.sub.1-4alkylindolyl, C.sub.1-4alkylpurinyl,
C.sub.1-4alkylcarbazolyl, C.sub.1-4alkylbenzimidazolyl, and
C.sub.1-4alkylisoxazolyl.
[0287] In one embodiment, R.sub.1 and/or R.sub.2 is a benzyl
optionally substituted with one or more of the following
substituents on the benzyl ring: X, --CH.sub.3, --NO.sub.2,
--OCH.sub.3, --CN, --CXH.sub.2, --CX.sub.2H, C.sub.2-C.sub.4 alkyl,
--CX.sub.3, --CH.sub.2(CX.sub.3), --CH(CX.sub.3).sub.2,
--C(CX.sub.3).sub.3, --C.sub.pX.sub.2p+1, --OCX.sub.3,
--OC.sub.pH.sub.2p+1, --OC.sub.pX.sub.2p+1, OR.sup.m, SR.sup.m,
NR.sup.mR.sup.n, NR.sup.mC(O)R.sup.n, SOR.sup.m, SO.sub.2R.sup.m,
C(O)R.sup.m, and C(O)OR.sup.m; R.sup.m and R.sup.n are
independently selected from H or a C.sub.1-C.sub.4 alkyl; and where
p is an integer from 2 to 20 and X is a halogen, including F, Cl,
Br, or I; preferably, F, Cl, or Br; more preferably, F or Cl.
XI. EXAMPLES
[0288] It should be understood that the description and examples
below are meant for purposes of illustration only and are not meant
to limit the scope of this disclosure. The examples below are meant
to illustrate the embodiments disclosed and are not to be construed
as being limitations to them. Additional compounds, other than
those described below, may be prepared by the following reaction
schemes or appropriate variations or modifications thereof.
Example 1. Synthesis of 2-Chlorobenzylamino-2-imidazoline
hydriodide
[0289] To a stirred solution of 2-methylthio-2-imidazoline
hydriodide (244 mg, 1.00 mMol) in dry dioxane (2.0 mL) was added
2-chlorobenzylamine (141 mg, 1.0 mMol). The reaction mixture was
stirred for 90 min at 70.degree. C. under argon. The solution was
cooled to room temperature, filtered on a sintered funnel, washed
with cold dioxane (2 mL) and dried under vacuum. The white solid
compound 4.HI (R.sub.2=2-chlorobenzyl) was obtained (242 mg, 72%)
and used without further purification.
Example 2. Synthesis of 2-Chlorobenzylamino-2-imidazoline
[0290] To a stirred solution of 2-chlorobenzylamino-2-imidazoline
hydriodide (242 mg, 0.72 mMol) in water (3 mL), was added 1.0 N
sodium hydroxide (2 mL) at 7.degree. C. The reaction mixture was
stirred for 30 min at 7.degree. C. under argon. After that
methylene chloride (5 mL) was added and the mixture stirred for
another 5 min. The reaction mixture was extracted with methylene
chloride (2.times.2.5 mL). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and evaporated. The resulting
free base (150 mg, 100%) was obtained as a viscous liquid and was
used for the next reaction without any further purification.
MS(ESI) 210 (M+H).
Example 3. Synthesis of Methyl-1-benzyl 4-oxo-3-piperidine
carboxylate (Compound (6))
[0291] To a stirred methyl-1-benzyl 4-oxo-3-piperidine carboxylate
hydrochloride (5.7 g, 20 mMol) in ethyl acetate (50 mL), was added
triethylamine (6 mL) at 7.degree. C. The reaction mixture was
stirred for 30 min at 7.degree. C. under an argon atmosphere. The
reaction mixture was extracted with ethyl acetate (2.times.50 mL)
and washed with water (50 mL). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and evaporated. The resulting
free base residue (5, R.sub.1=benzyl) as a viscous oil was used in
the next reaction without any further purification MS(ESI) 248
(M+H)
Example 4. Synthesis of ONC202 (Compound (14))
[0292] To a solution of 2-chlorobenzylamino-2-imidazoline (150 mg,
0.72 mMol), methyl 1-benzyl 4-oxo-3-piperidine carboxylate (5,
R.sub.1=benzyl) (195 mg, 0.79 mMol) in 1-butanol (2 mL) was added
PPTS (10 mg) and the mixture was stirred at room temperature for 48
h. After that the reaction mixture was refluxed at 125.degree. C.
to 130.degree. C. for 2 h. The solvents were removed under vacuum,
extracted with ethyl acetate (10 mL), and washed with saturated
sodium bicarbonate solution (2.times.10 mL) and water (10 mL). The
organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered
and evaporated. The crude free base was purified by RP HPLC
(10%-40% acetonitrile/water) to give ONC902 TFA salt as a white
solid (228 mg, 50% yield) MS(ESI) 407 (M+H).
[0293] The same process was used starting with different
benzylamines to prepare various analogs, e.g., ONC203, 204, 205,
206, 912, 210, 211, 212, 213, 214, 217, 218, 219, 220, 221, 222,
223, 224, 225, and 226.
Example 5. Synthesis of ONC207 (Compound (19))
[0294] To a suspension of 60% sodium hydride (3.5 g, 88 mMol) in
dry toluene (50 mL), dimethyl carbonate (4.32 g, 48.0 mMol) was
added dropwise in 0.5 h at room temperature under nitrogen. After
addition of a few drops of methanol,
1-tert-butoxycarbonyl-4-piperidone (4.8 g, 24 mMol) dissolved in
dry toluene (20 mL) was added dropwise to the reaction mixture
while stirring at 80.degree. C. over 1 h. The reaction mixture was
stirred for 3 h at the same temperature and then cooled to
0.degree. 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 layer was dried over
anhydrous sodium sulfate, and concentrated under reduced pressure.
The compound was dried in vacuum to give
methyl-1-tert-butoxycarbonyl-4-oxo-3-piperidine carboxylate (5.0 g,
80%). The compound obtained was carried to the next reaction
without any further purification.
[0295] 2-methybenzylamino-2-imidazoline (190 mg, 1 mMol), methyl
1-tert-butoxycarbonyl-4-oxo-3-piperidine carboxylate (315 mg, 1.1
mMol) in 1-butanol (2 mL) was added PPTS (10.0 mg) and the mixture
was stirred at room temperature for 48 h. After that the reaction
mixture was refluxed at 125.degree. C. to 130.degree. C. for 2 h.
The solvents were removed under vacuum, extracted with ethyl
acetate (10 mL), washed with saturated sodium bicarbonate solution
(2.times.10 mL) and water (10 mL). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, filtered and evaporated. The crude free
base was cleaved with 10% trifluoroacetic acid in dichloromethane,
purified by RP HPLC (10%-40% acetonitrile/water) to give ONC907
(262 mg, 50%) TFA salt as a white solid MS(ESI) 297 (M+H).
Example 6. Synthesis of ONC209 (Compound (21))
[0296] A mixture of ONC907 (100 mg, 0.2 mMol), phenylethyl bromide
(55.0 mg, 0.28 mMol) and potassium carbonate (150 mg, 1.0 mMol) in
N,N-dimethylformamide (3 mL) was heated to 70.degree. C. for 12 h.
The solvents were removed under vacuum, extracted with ethyl
acetate (10 mL), and washed with water (5 mL). The organic layer
was dried over anhydrous Na.sub.2SO.sub.4, filtered and evaporated.
The crude free base was purified by RP HPLC (10%-40%
acetonitrile/water) to give ONC209 (62 mg, 50%) TFA salt as a white
solid MS(ESI) 401 (M+H).
[0297] The same process was used starting with different halides to
give ONC215 and 214. Compounds 227, 228, 229, 230, 231, 232, 233,
234, 235, and 236 were prepared using an analogous process from
Examples 1 and 5 starting with different benzylamines. Then
treating the intermediate compound where R.sub.1 is H with
different halides as above.
[0298] Compound ONC216 was prepared from ONC215 by treatment with
TFA.
[0299] Compound (72) was prepared by reacting the precursor NH
compound prepared in analogy to Example 5 and treating it with
styrene oxide.
Example 7. Synthesis of ONC208 (Compound (20))
[0300] To a solution of 2-methylbenzylamino-2-imidazoline (190.0
mg, 1.0 mmol), methyl 1-methyl 4-oxo-3-piperidine carboxylate
(185.0 mg, 1.0 mMol) in 1-butanol (2.0 mL) was added PPTS (10.0 mg)
and the mixture was stirred at room temperature for 48 h. After
that the reaction mixture was refluxed at 125.degree. C. to
130.degree. C. for 2 h. The solvents were removed under vacuum,
extracted with ethyl acetate (10 mL), washed with saturated sodium
bicarbonate solution (2.times.10 mL) and water (10 mL). The organic
layer was dried over anhydrous Na.sub.2SO.sub.4, 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).
Example 8. Synthesis of ONC201 (Compound (1))
[0301] To a stirred 800 mL saturated NaHCO.sub.3 in a 2 L round
bottom flask, compound (3) (239.7 g, 0.845 mol, 1.6 equiv) was
added in portions. n-Butanol (500 mL) was added to the resulting
mixture, which was stirred for 30 min and then transferred to a
separating funnel. The organic phase, containing compound (4), was
separated and transferred to a 2 L three-neck round bottom flask
equipped with mechanical stirring, N2 inlet, a thermocouple, a
condenser and a Dean-Stark trap. To the contents of the flask,
Compound (5) (100 g, 0.528 mol, 1 equiv) and pyridinium
p-toluenesulfonate (PPTS) (6.63 gm 0.026 mol, 5 mol %) were added.
The resulting mixture was heated to reflux for 6 hours. Water in
the reaction mixture was separated into the Dean-Stark trap as
necessary. Refluxing temperature increased from 93.degree. C. to
118.degree. C. Reaction progress was monitored by HPLC. When the
peak area of compound (1) on HPLC remained constant with the
reaction time, the reaction was stopped.
Example 9. Synthesis of Di-Salt of ONC201 (Compound (1).2HCl)
[0302] Without isolation of the compound (1), the reaction mixture
from Example 8 was washed with water (500 mL) and diluted with
methyl tert-butyl ether (MTBE) (800 mL). The organic phase was
washed with water (500 mL.times.2) and transferred to a 3 L
three-neck round bottom flask equipped with mechanical stirring, Na
inlet, a thermocouple, a condenser and a Dean-Stark trap. While
agitating the reaction mixture, 1 N HCl in dioxane-MTBE solution
was added dropwise (4 N HCl in dioxane: 300 mL, 1.2 mol, 2.27
equiv; MTBE: 1200 mL) until no more solid precipitated out of the
reaction mixture upon addition of HCl. The reaction mixture was
heated to reflux at 60-65.degree. C. for 2 hours. Water was
separated into the Dean-Stark trap as necessary. Upon 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.degree. C. overnight (16 hours) to afford 200 g yellow
solid.
[0303] To a 2 L three-neck round bottom flask equipped with
mechanical stirring, Na inlet, a thermocouple and a condenser, the
above solid (200 g) was added, followed by ethanol (1000 mL). The
mixture was heated to reflux at 78.degree. C. for 2 hours. Upon
cooling to room temperature, the solid was filtered through a
sintered glass funnel and washed with ethanol (200 mL.times.3). The
wet solid was dried in the vacuum oven at 85.degree. C. for 3 days
until the residual solvent met specification. 120 g of compound (2)
was obtained as a white solid in a yield of 49%, with HPLC purity
99.7%.
Example 10. Activity of Imipridones
[0304] A number of imipridones were prepared based on the syntheses
above. For each compound, viability of human cancer cells at 72
hours post-treatment with the compound was measured. The change in
potency (relative to ONC201) was determined and shown in Table
3.
TABLE-US-00003 TABLE 3 RELATIVE POTENCY OF ONC201 ANALOGS Relative
No. Identifier R.sub.1 R.sub.2 Potency* 1 ONC201 CH.sub.2Ph
CH.sub.2-((2-CH.sub.3)--Ph) N/A 14 ONC202 CH.sub.2Ph
CH.sub.2(2-Cl--Ph) B 15 ONC203 CH.sub.2Ph CH.sub.2-(2-thienyl) C 16
ONC204 CH.sub.2Ph CH.sub.2CH.sub.2Ph C 17 ONC205 CH.sub.2Ph
CH.sub.2CH.sub.2(4-N-benzyl-piperazine) C 18 ONC206 CH.sub.2Ph
CH.sub.2-(2,4-di F--Ph) A 19 ONC207 H CH.sub.2-((2-CH.sub.3)--Ph) B
20 ONC208 CH.sub.3 CH.sub.2-((2-CH.sub.3)--Ph) B 21 ONC209
CH.sub.2CH.sub.2Ph CH.sub.2-((2-CH.sub.3)--Ph) B 32 ONC215
(CH.sub.2).sub.3--NH--BOC CH.sub.2-((2-CH.sub.3)--Ph) B 33 ONC216
(CH.sub.2).sub.3--NH.sub.2 CH.sub.2-((2-CH.sub.3)--Ph) B 41 ONC210
CH.sub.2Ph CH.sub.2-(3,5-di F--Ph) B 51 ONC211 CH.sub.2Ph
CH.sub.2-(3,4-di Cl--Ph) B 52 ONC212 CH.sub.2Ph
CH.sub.2-(4-CF.sub.3--Ph) A 53 ONC213 CH.sub.2Ph CH.sub.2-(3,4-di
F--Ph) A 54 ONC214 CD.sub.2C.sub.6D.sub.5
CH.sub.2-((2-CH.sub.3)--Ph) B 43 ONC217 CH.sub.2Ph
CH.sub.2(2-F--Ph) C 55 ONC218 CH.sub.2Ph CH.sub.2(2-CH.sub.3,
4-F--Ph) A 56 ONC219 CH.sub.2Ph CH.sub.2-(2,4-di Cl--Ph) A 57
ONC220 CH.sub.2Ph CH.sub.2-((4-OCH.sub.3)--Ph) A 35 ONC222
CH.sub.2Ph CH.sub.2-(3-isoxazolidinyl) C 36 ONC224 CH.sub.2Ph
CH.sub.2CH.sub.2-(4-morpholinyl) A 38 ONC221 H
CH.sub.2-(4-CF.sub.3--Ph) A 72 ONC225 CH.sub.2Ph CH.sub.2-(2-F,
4-CF.sub.3--Ph) A 37 ONC223 CH.sub.2Ph CH.sub.2-(4-CH.sub.3--Ph) A
34 ONC226 CH.sub.2Ph CH.sub.2-(3-pyridinyl) A 77 ONC231
CH.sub.2-3-pyridyl CH.sub.2-(4-CF.sub.3--Ph) A 78 ONC232
CH.sub.2-4-methyl-2-thiazolyl CH.sub.2-(4-CF.sub.3--Ph) B 79 ONC233
CH.sub.2-2-pyrazinyl CH.sub.2-(4-CF.sub.3--Ph) B 81 ONC234
CH.sub.2-(3,4-di Cl--Ph) CH.sub.2-(4-CF.sub.3--Ph) A 83 ONC236
CH.sub.2-3-thienyl CH.sub.2-(4-CF.sub.3--Ph) A 84 ONC237
CH.sub.2CH(OH)Ph CH.sub.2-(4-CF.sub.3--Ph) C 73 ONC227
CH.sub.2-(4-CF.sub.3--Ph) CH.sub.2-(4-CF.sub.3--Ph) B 74 ONC228
CH.sub.2-(4-F--Ph) CH.sub.2-(4-CF.sub.3--Ph) A 75 ONC229
CH.sub.2-(4-OCH.sub.3--Ph) CH.sub.2-(4-CF.sub.3--Ph) B 76 0NC230
4-F--Ph-4-oxobutyl CH.sub.2-(4-CF.sub.3--Ph) A *Relative to the
potency of ONC201; A Indicates a potency increase of >2-fold of
ONC201; B Indicates potency that is within 2-fold of ONC201; and C
Indicates a potency decrease of >2-fold of ONC201.
##STR00027##
[0305] The IC.sub.50 of ONC201 and ONC212 (5 nM-5 .mu.M, 72 h) upon
treatment of several acute myeloid leukemia (AML) cell lines (n=3)
were determined and shown below in Table 11.
TABLE-US-00004 TABLE 11 AML cell line ONC201 IC.sub.50 (.mu.M)
ONC212 IC.sub.50 (.mu.M) MV411 3.25 0.01 HL60 >5 0.21 MOLM14
3.92 0.01
[0306] Cell viability of MV411 AML cells treated with ONC212 and
cytarabine (5 nM-5 .mu.M, 24 h) (n=3) was measured (FIG. 29A).
Furthermore, cell viability MOLM14, MV411 AML cells, MRCS lung
fibroblasts and Hs27a bone marrow cells treated with ONC212 (5 nM-5
.mu.M, 72 h) (n=3) was measured (FIG. 29B). Cell viability of
MOLM14 and MV411 AML cells treated with ONC212 (250 nM) for 4, 8,
24, 48, 72 and 96 h was measured. ONC212 medium was replaced by
fresh medium at these time points and cell viability was determine
at 96 h for all samples. (n=2) (FIG. 29C).
[0307] In addition, a single dose of compound (52) (ONC212) by oral
or intraperitoneal administration to human colon cancer
xenograft-bearing mice resulted in significant reduction of tumor
volume compared to vehicle-treated control cohorts (FIG. 24).
Compound (52) has a wide therapeutic window, as it is well
tolerated at doses at least up to 225 mg/kg in mice.
[0308] Furthermore, ONC212 demonstrated efficacy in
ONC201-resistant AML xenograft model (FIG. 30). MV411 AML cells
(5.times.10.sup.6) were subcutaneously implanted in the flanks of
athymic nude. ONC212 and ONC201 were administered orally (PO) as
indicated. Tumor volume (A and B) and body weight (C) (n=10) was
measured on indicated days. * represents p<0.05 relative to
vehicle.
[0309] ONC212 efficacy in AML was evaluated in vitro and was upto
400 fold more potent compared to ONC201 (Table 11). ONC212 was also
efficacious in AML cells resistant to standard of care cytarabine
(FIG. 29A). Despite robust improvement in efficacy ONC212 maintains
a wide therapeutic window in vitro and is non-toxic to normal cells
at efficacious concentrations (FIG. 29B). An 8 hr exposure of
ONC212 at 250 nM was sufficient to cause robust reduction in cell
viability in MOLM14 and MV411 AML cells (FIG. 29C). At least 24-48
h exposure was required with ONC201 for efficacy.
[0310] ONC212 efficacy was determined in a leukemia xenograft model
with MV411 AML cells resistant to standard-of-care cytarabine (FIG.
30). ONC212 50 mg/kg significantly reduced leukemia xenograft tumor
growth with oral weekly administration while ONC201 was not
efficacious in this model at similar doses (FIG. 30A). Interesting,
biweekly ONC212 dosing with 25 mg/kg and weekly/biweekly dosing
with 5 mg/kg was not efficacious (FIG. 30B). None of these ONC212
administration regimens were associated with body weight loss (FIG.
30C) or gross observations.
[0311] ONC212 25 mg/kg represents NOAEL in mouse and rat non-GLP
oral single dose studies which is also the efficacious dose in
mouse xenograft studies. ONC212 is approximately 10 fold more toxic
compared to ONC201 (NOAEL 225 mg/kg in rat non-GLP oral single dose
study).
##STR00028##
[0312] ONC206 demonstrated efficacy in a Ewing's sarcoma xenograft
model (FIG. 31). MHH-ES-1 Ewing's sarcoma cells (5.times.10.sup.6)
were subcutaneously implanted in the flanks of athymic nude mice.
ONC206 (PO) and methotrexate (IV) were administered on day 1 and
day 13 as indicated. Tumor volume (FIG. 31A) and body weight (FIG.
31B) (n=4) was measured on indicated days.
[0313] In addition, the IC.sub.50 of ONC201 and ONC206 (5 nM-5
.mu.M, 72 h) upon treatment of several cell lines (n=3) were
determined and shown below in Table 11.
TABLE-US-00005 TABLE 12 ONC201 ONC206 Cell line IC.sub.50 (.mu.M)
IC.sub.50 (.mu.M) MV411 (AML) 3.25 0.2 K562 (CML) >5 0.22 MOLM14
(AML) 3.92 0.27 MHH-ES-1 (Ewing's sarcoma) 5.65 0.61 HFF (Normal
Fibroblast) >5
[0314] ONC206 showed up to 20 fold improvement compared to ONC201
in in vitro potency with no in vitro toxicity to normal cells at
therapeutic doses (Table 12). With ONC206, only 2-fold increased
toxicity (NOAEL 125 mg/kg) was noted overall relative to ONC201
(NOAEL 225 mg/kg) in rat non-GLP oral single dose study. In vivo
efficacy in Ewing's sarcoma model with no toxicity (FIG. 31).
ONC206 efficacy was comparable to chemotherapy methotrexate, but
chemotherapy was associated with body weight loss.
##STR00029##
[0315] In vitro profiling of GPCR activity using a hetereologous
reporter assay for arrestin recruitment, a hallmark of GPCR
activation, indicated that ONC213 selectively targets DRD2/3 and
GPR132/91 (FIG. 32). Dual targeting of DRD2/3 and GPR132/91
represents a novel strategy for anti-cancer efficacy without
toxicity. ONC213 is a DRD2/3 inhibitor and a GPR132/91 agonist.
DRD2/3 potency of ONC213 is more than ONC201 but less than ONC206.
GPR132 potency of ONC213 is less than ONC212. Specifically, ONC213
demonstrated in vitro anti-cancer potency in HCT116/RPMI8226 cancer
cells similar to ONC212, but in vitro toxicity to normal cells was
reduced compared to ONC212 (FIG. 33). The safety profile of ONC213
confirmed in mouse MTD study with NOAEL 75 mg/kg three times that
of ONC212 (25 mg/kg). The GPR91 agonist activity of ONC213 provides
an opportunity for immunology, immune-oncology and hematopoietic
applications (Nature Immunology 9:1261 (2008); J Leukoc Biol.
85(5):837 (May 2009)).
##STR00030##
[0316] In vitro profiling of GPCR activity using a hetereologous
reporter assay for arrestin recruitment, a hallmark of GPCR
activation, indicated that ONC237 selectively targets DRD5 and
GPR132 (FIG. 34). ONC237 is a GPR132 agonist and DRD5 antagonist
and has reduced anticancer efficacy (IC.sub.50 31.2 .mu.M) compared
to ONC201. This data show that combining GPR132 agonist activity
with DRD5 (D1-like dopamine receptor) antagonist activity results
in poor anti-cancer effects compared to ONC213 which combines
GPR132 agonist and DRD2/3 antagonist activity.
##STR00031##
[0317] In vitro profiling of GPCR activity using a hetereologous
reporter assay for arrestin recruitment, a hallmark of GPCR
activation, indicated that ONC236 is a highly selective GPR132
agonist (FIG. 35). ONC236 has anticancer efficacy (IC.sub.50 88 nM)
comparable to ONC212 (10 nM) better than ONC206/ONC201,
completeness of response is better than ONC201 but not ONC212 in
HCT116 cells.
##STR00032##
[0318] In vitro profiling of GPCR activity using a hetereologous
reporter assay for arrestin recruitment, a hallmark of GPCR
activation, indicated that ONC234 is a broad spectrum and potent
GPCR targeting small molecule (FIGS. 36 and 38). ONC234 hits
several GPCRs including activity as an antagonist activity for
adrenergic, histamine, serotonin, CHRM, CCR, DRD2/5 receptors, as
well as CXCR7 agonist activity. ONC236 has anticancer efficacy
(IC.sub.50 234 nM) similar to ONC206, completeness of response same
as ONC212, and better than ONC201 in HCT116 cells.
ONC201 Linear Isomer (TIC-10)
##STR00033##
[0320] In vitro profiling of GPCR activity using a hetereologous
reporter assay for arrestin recruitment, a hallmark of GPCR
activation, indicated that the ONC201 linear isomer (TIC-10) is a
CXCR7 agonist (FIG. 37). CXCR7 agonists can be used for liver
regeneration and preventing/treating fibrosis, such as liver
fibrosis (Nature 505:97 (2014)). Fibrosis is the formation of
excess fibrous connective tissue in an organ or tissue, including
as a result of wound healing. Examples of fibrosis includes,
pulmonary fibrosis, including cystic fibrosis and idiopathic
pulmonary fibrosis; radiation-induced lung injury following
treatment for cancer; liver fibrosis (cirrhosis); heart fibrosis,
including atrial fibrosis, endomyocardial fibrosis, and old
myocardial infarction; glial scar; arthrofibrosis; Crohn's Disease;
dupuytren's contracture; keloids; mediastinal fibrosis;
myelofibrosis; Peyronie's disease; nephrogenic systemic fibrosis;
progressive massive fibrosis; retroperitoneal fibrosis;
scleroderma/systemic sclerosis; and adhesive capsulitis.
Example 11. GPCR Antagonism of ONC201
[0321] ONC201 was evaluated in a whole cell, functional assay of
.beta.-Arrestin G protein-coupled receptor (GPCR) activity that
directly measures dopamine receptor activity by detecting the
interaction of .beta.-Arrestin with the activated GPCR that serves
as a reporter. For each dopamine receptor (DRD1, DRD2S, DRD2L,
DRD3, DRD4, and DRD5), cell lines overexpressing reporter
constructs were expanded from freezer stocks. Cells were seeded in
a total volume of 20 .mu.L into white walled, 384-well microplates
and incubated at 37.degree. C. prior to testing, with antagonist
followed by agonist challenge at the EC.sub.80 concentration.
Intermediate dilution of sample stocks was performed to generate
5.times. sample in assay buffer. 3.5 .mu.L of 5.times. sample was
added to cells and incubated at 37.degree. C. or room temperature
for 30 minutes. Vehicle concentration was 1%. 5 .mu.L of 6.times.
EC.sub.80 agonist in assay buffer was added to cells and incubated
at 37.degree. C. or room temperature for 90 or 180 minutes prior to
assay readout. % Antagonism was calculated using the following
formula %: Antagonism=100%.times.(1-(mean RLU of test sample-mean
RLU of vehicle control)/(mean RLU of EC.sub.80 control-mean RLU of
vehicle control).
Example 12: Selective Antagonism of DRD2 by ONC201
[0322] ONC201 is a first-in-class small molecule discovered in a
phenotypic screen for p53-independent inducers of tumor selective
proapoptotic pathways. Oral ONC201 is being evaluated as a new
therapeutic agent in five early phase clinical trials for select
advanced cancers based on pronounced efficacy in aggressive and
refractory tumors and excellent safety.
[0323] In this Example, the prediction and validation of selective
direct molecular interactions between ONC201 and specific dopamine
receptor family members are reported. Experimental GPCR profiling
indicated that ONC201 selectively antagonizes the D2-like, but not
D Hike, dopamine receptor subfamily Reporter assays in a
heterologous expression system revealed that ONC201 selectively
antagonizes both short and long isoforms of DRD2 and DRD3, with
weaker potency for DRD4 and no antagonism of DRD1 or DRD5.
Increased secretion of prolactin is a clinical hallmark of DRD2
antagonism by several psychiatric medications that potently target
this receptor. ELISA measurements in peripheral blood of patients
treated with ONC201 in the first-in-human trial with advanced solid
tumors determined that 10/11 patients evaluated exhibited induction
of prolactin (mean of 2-fold).
[0324] Using the TCGA database, the D2-like dopamine receptor
subfamily, particularly DRD2, was found to be prevalent and
selectively overexpressed in several malignancies. Preclinical
reports show that DRD2 inhibition imparts antitumor efficacy,
without killing normal cells, via induction of ATF4/CHOP and
inhibition of Akt and ERK signaling that are all attributes of
ONC201.
Methods
[0325] ONC201 dihydrochloride was obtained from Oncoceutics. Kinase
inhibition assays for the kinome were performed as described (see
Anastassiadis et al., Nat Biotech 29:1039 (2011)). GPCR arrestin
recruitment and cAMP modulation reporter assays were performed as
described (see McGuinness et al., Journal of Biomolecular Screening
14:49 (2009)). PathHunter.TM. (DiscoveRx) .beta.-arrestin cells
expressing one of several GPCR targets were plated onto 384-well
white solid bottom assay plates (Corning 3570) at 5000 cells per
well in a 20 .mu.L volume in an appropriate cell plating reagent.
Cells were incubated at 37.degree. C., 5% CO.sub.2 for 18-24 h.
Samples were prepared in buffer containing 0.05% fatty-acid free
BSA (Sigma). For agonist mode tests, samples (5 .mu.L) were added
to pre-plated cells and incubated for 90 minutes at 37.degree. C.,
5% CO.sub.2. For antagonist mode tests, samples (5 .mu.L) were
added to pre-plated cells and incubated for 30 minutes at
37.degree. C., 5% CO.sub.2 followed by addition of EC.sub.80
agonist (5 .mu.L) for 90 minutes at 37.degree. C., 5% CO.sub.2. For
Schild analysis, samples (5 .mu.L) were added to pre-plated cells
and incubated for 30 minutes at 37.degree. C., 5% CO.sub.2 followed
by addition of serially dliuted agonist (5 .mu.L) for 90 minutes at
37.degree. C., 5% CO.sub.2. Control wells defining the maximal and
minimal response for each assay mode were tested in parallel.
Arrestin recruitment was measured by addition of 15 .mu.L
PathHunter Detection reagent and incubated for 1-2 h at room
temperature and read on a Perkin Elmer Envision Plate Reader. For
agonist and antagonist tests, data was normalized for percent
efficacy using the appropriate controls and fitted to a sigmoidal
dose-response (variable slope),
Y=Bottom+(Top-Bottom)/(1+10{circumflex over ( )}((Log
EC.sub.50-X)*HillSlope)), where X is the log concentration of
compound. For Schild analysis, data was normalized for percent
efficacy using the appropriate controls and fitted to a
Gaddum/Schild EC.sub.50 shift using global fitting, where
Y=Bottom+(Top-Bottom)/(1+10{circumflex over ( )}((Log
EC-X)*HillSlope)), Antag=1+(B/(10{circumflex over (
)}(-1*pA2))){circumflex over ( )}SchildSlope and Log
EC=Log(EC.sub.50*Antag). EC.sub.50/IC.sub.50 analysis was performed
in CBIS data analysis suite (Cheminnovation) and Schild analysis
performed in GraphPad Prism 6.0.5.
Results
[0326] ONC201 is a small molecule in phase II clinical trials for
select advanced cancers. It was discovered in a phenotypic screen
for p53-independent inducers of the pro-apoptotic TRAIL pathway.
Although the contribution of ONC201-induced ATF4/CHOP upregulation
and inactivation of Akt/ERK signaling (Allen et al., Science
translational medicine 5, 171ra117-171ra117 (2013)) to its
anti-cancer activity has been characterized, its molecular binding
target has remained elusive.
[0327] In vitro profiling of GPCR activity using a hetereologous
reporter assay for arrestin recruitment, a hallmark of GPCR
activation, indicated that ONC201 selectively antagonizes the
D2-like (DRD2/3/4), but not D1-like (DRD1/5), dopamine receptor
subfamily (FIGS. 4B and 5A). Antagonism of adrenoceptor alpha
receptors or other GPCRs was not observed under the evaluated
conditions. Among the DRD2 family, ONC201 antagonized both short
and long isoforms of DRD2 and DRD3, with weaker potency for DRD4.
Further characterization of ONC201-mediated antagonism of arrestin
recruitment to DRD2L was assessed by a Gaddum/Schild EC.sub.50
shift analysis, which determined a dissociation constant of 2.9
.mu.M for ONC201 that is equivalent to its effective dose in many
human cancer cells (FIG. 4C). Confirmatory results were obtained
for cAMP modulation in response to ONC201, which is another measure
of DRD2L activation (FIG. 4D). The ability of dopamine to reverse
the dose-dependent antagonism of up to 100 .mu.M ONC201 suggests
direct, competitive antagonism of DRD2L (FIGS. 5B and 5C). In
agreement with the ONC201 specificity predicted by BANDIT, no
significant interactions were identified between ONC201 and nuclear
hormone receptors, the kinome, or other drug targets of
FDA-approved cancer therapies (FIGS. 5D and 5E). Interestingly, a
biologically inactive constitutional isomer of ONC201 (Wagner et
al., Oncotarget 5:12728 (2014)) did not inhibit DRD2L, suggesting
that antagonism of this receptor could be linked to its biological
activity (FIG. 5F). In summary, these studies establish that ONC201
selectively antagonizes the D2-like dopamine receptor subfamily,
which appears to be a promising therapeutic target in oncology, and
ONC201 is the first compound to exploit this treatment paradigm in
several ongoing Phase II clinical studies.
Example 13: Shotgun Mutagenesis Epitope Mapping of DRD2
[0328] Shotgun Mutagenesis uses a high-throughput cellular
expression technology to express and analyze large libraries of
mutated target proteins within eukaryotic cells. Every residue in a
protein is individually mutated to an alanine, or other specified
residue, to assay changes in function. Proteins are expressed
within standard mammalian cell lines, therefore even difficult
proteins that require eukaryotic translational or
post-translational processing can be mapped.
[0329] First, conditions were evaluate and identified for screening
the DRD2 antagonist ONC201 with wild-type DRD2 using the Shotgun
Mutagenesis screening assay. Then, a DRD2 Ala-scan library was
prepared and the residues critical for ONC201 binding were mapped
at single amino acid resolution using Shotgun Mutagenesis
technology.
DRD2 Shotgun Mutagenesis Library:
[0330] Parental plasmid: DRD2 Library size: 442 mutant clones
(complete protein) Mutation Strategy: Alanine scan mutagenesis Cell
type: HEK-293T Screening Assay: Calcium flux
Epitope Tag: C-terminal V5/HIS6
[0331] Parental Construct: DNA encoding the full-length human DRD2
(Accession No: NP_000786.1; MDPLNLSWYD DDLERQNWSR PFNGSDGKAD
RPHYNYYATL LTLLIAVIVF GNVLVCMAVS REKALQTTTN YLIVSLAVAD LLVATLVMPW
VVYLEVVGEW KFSRIHCDIF VTLDVMMCTA SILNLCAISI DRYTAVAMPM LYNTRYSSKR
RVTVMISIVW VLSFTISCPL LFGLNNADQN ECIIANPAFV VYSSIVSFYV PFIVTLLVYI
KIYIVLRRRR KRVNTKRSSR AFRAHLRAPL KGNCTHPEDM KLCTVIMKSN GSFPVNRRRV
EAARRAQELE MEMLSSTSPP ERTRYSPIPP SHHQLTLPDP SHHGLHSTPD SPAKPEKNGH
AKDHPKIAKI FEIQTMPNGK TRTSLKTMSR RKLSQQKEKK ATQMLAIVLG VFIICWLPFF
ITHILNIHCD CNIPPVLYSA FTWLGYVNSA VNPIIYTTFN IEFRKAFLKI LHC (SEQ ID
NO: 1) was subcloned into a mammalian high-expression vector. This
parental construct was sequence-verified and then validated for
mammalian cell expression by detection of calcium flux in response
to dopamine DNA yields from plasmid preparations have been
validated for high-throughput processing.
[0332] Assay Set-up: A DRD2-specific calcium flux assay was
successfully optimized for DRD2 expressed in human cells. An
agonist dose-response assay was used to identify a suitable
dopamine concentration for use in optimizing the inhibition of
DRD2-specific calcium flux by antagonist ONC201. Subsequent
dose-response inhibition assays identified a concentration of
ONC201 that inhibited the DRD2 dopamine response by >95%.
Calcium Flux Assay Optimization:
[0333] Receptor Activity Assay. DRD2 activity was assessed using a
published GPCR assay (Greene, T. A. et al., (2011) PLoS One 6,
e20123). Briefly, HEK-293T cells were transfected with expression
constructs for wild-type DRD2 or a negative control GPCR, in
384-well format. After 22 hr, calcium flux experiments were
performed over a range of dopamine concentrations (300 pM-100 nM),
using a Flexstation II-384 fluorescence reader (Molecular Devices).
Data sets were analyzed and represented as percentage over baseline
signal using Prism 5.0 software (GraphPad Software, Inc).
[0334] For cells expressing DRD2, but not a control GPCR, addition
of dopamine resulted in increases in cellular calcium flux,
measured as increased fluorescence. A dose response plot of the
fluorescence peak height versus dopamine concentration demonstrated
the strong dopamine-induced calcium flux (EC.sub.50=0.45 nM) in
cells expressing DRD2, but not the control GPCR. This suggested
that the calcium flux assay could be used to test for ONC201
inhibition.
DRD2 Calcium Flux Inhibition Assay Optimization
[0335] Following identification of the EC.sub.50 for dopamine in
the calcium flux assay, ONC201 inhibition of DRD2-specific calcium
flux was investigated at several dopamine concentrations. Using 1
nM dopamine (>2-fold higher than the dopamine EC.sub.50) with a
range of ONC201 concentrations (1 nM to 100 .mu.M), ONC201
inhibition of dopamine-induced DRD2 calcium flux was observed at
the highest concentrations tested (FIG. 9A), with complete
inhibition by 100 .mu.M ONC201 (IC.sub.50=21.5 .mu.M). Inhibition
of calcium flux by 100 .mu.M ONC201 was not the result of a broad
inhibition of GPCRs or of a non-specific effect on cells since
ONC201 had no effect on the calcium flux activity of cells
expressing a control GPCR (FIG. 9B).
[0336] Analysis of a number of replicate values obtained for
inhibition of DRD2 calcium flux by 100 .mu.M ONC201 indicated a
robust assay, with a Z' value of 0.61. The Z' value is a
measurement of assay quality, calculated from the means and
standard deviations obtained for replicate determinations of
calcium flux obtained with or without ONC201.
Comparison of DRD2 Inhibitors.
[0337] The ONC201 inhibition of DRD2 was compared to that by the
DRD2 antagonists spiperone and domperidone (FIG. 10), which have
been described as inhibiting DRD2 at concentrations lower than the
100 .mu.M required for inhibition by ONC201. These antagonists were
screened at concentrations between 100 pM and 1 .mu.M, and both
showed complete inhibition of dopamine-induced calcium flux, with
spiperone having an IC.sub.50=19 nM, and domperidone an
IC.sub.50=47 nM. These values were consistent with previous
characterizations and demonstrate that the relatively high
IC.sub.50 obtained for ONC201 (21.5 .mu.M) does not result from the
use of a calcium flux assay to measure DRD2 activity.
[0338] Optimal screening conditions were determined for ONC201
inhibition of DRD2-specific calcium flux in response to dopamine.
These conditions give a robust response to dopamine, this response
is reduced by >95% by addition of ONC201 to 100 .mu.M, and the
assay demonstrated low variability between replicates. These data
indicate that the selected conditions are suitable for successful
high-throughput screening. Further screening of the DRD2 mutation
library was at a dopamine concentration of 1 nM and an ONC201
concentration of 100 .mu.M.
Screening the DRD2 Alanine-Scan Library for Response to
Dopamine.
[0339] The DRD2 alanine-scan mutation library (and with alanines
changed to serines) comprised 442 clones, covering residues 2-443
of the DRD2 protein, 100% of target residues. The DRD2 mutation
library was first screened by calcium flux assay with dopamine (1
nM) in the absence of ONC201 to identify residues whose mutation
diminished dopamine-induced calcium flux. We identified 28 amino
acid residues that were critical for dopamine-induced DRD2 flux
(FIG. 11).
[0340] Residues were identified from the analysis are listed in
Table 4 and shown in FIG. 11. Clones were considered to be
deficient for calcium flux if they demonstrated flux values less
than 2 standard deviations below the average calcium flux value
(AV-2SD) for the entire library.
TABLE-US-00006 TABLE 4 DRD2 RESIDUES CRITICAL FOR DOPAMINE-INDUCED
CALCIUM FLUX Calcium Flux Calcium Flux Mutation % WT Mutation % WT
C182A 0 S7A 15 I184A 0 W386A 15 S197A 0 S121A 16 T119A 1 I394A 16
S193A 1 E248A 19 D80A 3 V190A 20 R132A 3 Y199A 20 D114A 4 C107A 20
H393A 4 S419A 20 F198A 10 F189A 22 V83A 10 I122A 23 I377A 11 T205A
24 Y416A 12 N23A 25 C118A 14 L125A 25 I128A 27
Screening the DRD2 Alanine-Scan Library for ONC201 Inhibition of
Dopamine-Induced Signaling Identified Residues Required for
Inhibition by ONC201.
[0341] To identify residues important for the inhibition of DRD2 by
ONC201, the DRD2 alanine-scan mutation library was screened by the
calcium flux assay for the ability to respond to dopamine in the
presence of an inhibiting concentration of ONC201, using dopamine
at 1 nM and ONC201 at 100 .mu.M. Eight residues critical for ONC201
inhibitory activity were identified (FIG. 12). All residues
identified by this screen showed high calcium flux with dopamine
alone (Table 5). Clones were considered to be critical for
inhibition by ONC201 at 100 .mu.M if they demonstrated flux values
greater than 2 standard deviations above the average calcium flux
value (AV+2SD) for the entire library. Also shown in Table 5 for
these critical clones are the calcium flux values obtained from
similar experiments performed with 250 .mu.M ONC201 or without
ONC201 (dopamine 1 nM), and in addition the % conservation of the
critical residues across the 5 DRD receptors, with the residues
found in each receptor.
TABLE-US-00007 TABLE 5 DRD2 RESIDUES CRITICAL FOR ONC201 INHIBITION
OF DOPAMINE-INDUCED CALCIUM FLUX Calcium Flux as a % of flux with
WT DRD2 (100) ONC201 ONC201 Dopamine DRD % DRD Mutation 100 .mu.M
250 .mu.M 1 nM Conservation 1 2 3 4 5 I397A 122 89 105 20 P I T A P
E95A 97 39 123 100 E E E E E V91A 94 58 119 40 K V V F K Y192A 85
11 64 60 S Y Y Y S V196A 79 22 119 40 I V V C I A177S 77 26 85 40 A
A T V D T165A 67 28 92 20 L T A A L L81A 63 20 83 100 L L L L L
[0342] Since the average inhibition by 100 .mu.M ONC201 across the
library was approximately 75%, we also conducted a screen at 250
.mu.M ONC201 to determine if critical residues would be the same at
higher levels of inhibition. Under this condition dopamine-induced
calcium flux was inhibited by approximately 93%, and the previously
identified residues V91, E95, and 1397 were also critical for
inhibition at 250 .mu.M ONC201 (Table 5), using the same criteria
of flux values greater than 2 standard deviations above the average
calcium flux value (AV+2SD) for the library.
CONCLUSIONS
[0343] In initial screens of the DRD2 alanine-scan mutation library
by dopamine-induced calcium flux assay, 28 mutations greatly
decreased calcium flux, identifying resides critical for DRD2
function. As found in a similar analysis of the GPCR CXCR4, the
critical residues were distributed throughout the protein, in the
predicted dopamine binding pocket, the transmembrane regions and in
the cytoplasmic exposed portion of DRD2. These 28 residues are
critical for either dopamine binding, signal transduction through
the transmembrane domains, or G protein coupling. A detailed
analysis comparable to that performed for CXCR4, as well as the
structural analysis of the DRD3-eticlopride structure (Chien et
al., 2010), can be used to assign specific function to each DRD2
critical residue.
[0344] To identify residues important for the inhibition of DRD2 by
ONC201, the DRD2 alanine-scan mutation library was screened by
calcium flux assay with dopamine and 100 .mu.M ONC201. These
screens identified 8 residues as critical for ONC201 inhibition of
DRD2-dependent dopamine-induced calcium flux--L81, V91, E95, T165,
A177, Y192, V196, and 1397. Residues V91, E95, and 1397 were also
identified as critical for resistance to DRD2 inhibition by 250
.mu.M ONC201, suggesting that they are key ONC201-interacting
residues. These residues define a relatively large ligand binding
site, which is not unexpected due to the larger size of ONC201
compared to dopamine and eticlopride. The locations of these
residues are generally consistent with a role in mediating ONC201
inhibition of DRD2-dependent dopamine-induced calcium flux.
Residues critical for inhibition of a GPCR taste receptor by
probenecid were previously identified (Greene et al., 2011), with
the location of the residues consistent with a non-competitive
mechanism of inhibition. In contrast, the residues identified here
for DRD2 are consistent with competitive inhibition by ONC201 at
the dopamine binding site. When modeled on the structure of
homologous receptor DRD3, the majority of the residues identified
surround the binding pocket containing a co-crystallized antagonist
eticlopride, with 5 of the 8 identified residues conserved between
DRD2 and DRD3. Two of the residues appear to more distal from the
putative binding site (A177 and L81) and may affect ONC201 binding
in a more allosteric fashion. Additional residues that contribute
to ONC201 inhibition may be identified using DRD2 agonists with
structures distinct from dopamine
Example 14: Determination of the Association & Dissociation
Rate Constants of Unlabeled ONC201 Dihydrochloride on the Human
D.sub.2S Receptor
[0345] In this Example, the kon/koff rates of unlabeled ONC201
dihydrochloride on the D.sub.2S receptor was determined. The
kon/koff rate estimation was performed by competitive ligand
binding according to the method described in: M. R. Dowling &
S. J. Charlton (2006) Brit. J. Pharmacol. 148:927-937 and H. J.
Motulsky & L. C. Mahan (1984) Mol. Pharmacol. 25:1-9. Referring
to this method, the kon/koff rates of the unlabeled test compounds
were calculated from its Ki value (competition binding) and its
effect on the binding kinetics of the radioligand (competition
kinetics).
[0346] First, the IC.sub.50 and Ki values of ONC201
dihydrochloride, and selection of the adequate compound
concentrations for the competition kinetics experiment, were
determined. Then, the kon and koff rate constants of the
radioligand ([.sup.3H]Methylspiperone) was determined. Finally, the
kon and koff rate constants of the unlabeled ONC201 dihydrochloride
was determined. ONC201 dihydrochloride was tested at 8
concentrations in duplicate (n=2) in the competition binding assay,
and the IC.sub.50 and Ki values were determined.
[0347] The reference compound, (+) Butaclamol, and the test
compound, ONC201-2HCL, successfully competed for
[.sup.3H]Methylspiperone, with IC.sub.50 values of 2.5 nM and 21
.mu.M, respectively. Previously, the compound ONC201-2HCL yielded a
similar IC.sub.50 value of 16 .mu.M. For the competition binding
assay, the following 6 concentrations of ONC201-2HCL were selected:
5/10/20/40/60/80 .mu.M.
[0348] The binding kinetics of [.sup.3H]Methylspiperone on the
D.sub.2S receptor was determined. For this,
[.sup.3H]Methylspiperone (at one concentration of 0.3 nM) was
incubated with the D.sub.2S receptor membranes for 12 different
incubation times to measure the association rate. The non-specific
binding was measured with Butaclamol (10 .mu.M) for each incubation
time. The dissociation was initiated by addition of an excess of
Butaclamol (10 .mu.M) after 60 minutes incubation of
[.sup.3H]Methylspiperone (0.3 nM) with the D.sub.2S receptor
membranes, and the signal decrease was measured after 12 different
incubation times. The experiment was performed in triplicate (n=3)
with incubation times adjusted to
0/30/60/80/120/180/240/300/360/420/480 minutes and 24 hours for the
association and 2/5/8/10/15/20/25/30/40/60/120/180 minutes for the
dissociation kinetics.
[0349] [3H]Methylspiperone displayed a k.sub.on value of
2.3.times.10.sup.8 M.sup.-1 min.sup.-1 and a k.sub.off value of
0.009506 min.sup.-1 (and thus a t.sub.1/2 value of 73 minutes) on
the D.sub.2S receptor. The K.sub.d calculated from the results of
the association/dissociation experiment (0.04 nM) is in the same
range as compared to the K.sub.d observed in the saturation
experiment (0.15 nM), thereby validating the experiment.
[0350] The effect of the unlabeled ONC201-2HCl at six
concentrations on the association kinetics of
[.sup.3H]Methylspiperone (0.3 nM) was tested. The non-specific
binding was measured with Butaclamol (10 .mu.M). The same 12
incubation times as above were used:
2/5/8/10/15/20/25/30/40/60/120/180 minutes. A measurement in the
absence of compound was performed as negative control.
[0351] ONC201-2HCl displayed a k.sub.on value of 4.1.times.10.sup.5
M.sup.-1 min.sup.-1 and a k.sub.off value of 1.32 min.sup.-1 (and
thus a t.sub.1/2 value of 0.53 minutes) on the D.sub.2S receptor.
The K, calculated from the results of the association/dissociation
experiment (3.2 .mu.M) is in the same range as compared to the K,
observed in the saturation experiment (7 .mu.M), thereby validating
the experiment. In conclusion, ONC201-2HCl displays a much slower
association and a much faster dissociation as compared to
[.sup.3H]Methylspiperone.
Example 15: Bactericidal Activity of Imipridones
Materials and Methods
[0352] Test material: ONC201 dihydrochloride; Control:
Microcrystalline Cellulose.
[0353] Method: Harmonized EP/USP Microbial Examination of
Nonsterile Products (Current USP <61>/<62>).
Results
TABLE-US-00008 [0354] TABLE 6 VERIFICATION OF THE INOCULUM RECOVERY
CONTROL AND MICROBIAL ENUMERATION TEST 1:300 with TSB Indicator
Organisms Count Mod Dilution Ec Sa Pa Bs Ca (TSA) Ab (TSA) CA (SDA)
Ab (SDA) Inoculum 27 31 28 52 48 21 52 20 434019 N/A 0 24 48 51 18
46 19
TABLE-US-00009 TABLE 7 THE VALIDATION FOR SPECIFIED MICROORGANISMS
Sample BTGN Ec Pa Sa Ca 1:300 with P P P F P TSB Mod Dilution P =
Pass F = Fail NA = Not Applicable; Ec = Escherichia coli ATCC#
8739; Pa = Pseudomonas aeruginosa ATCC# 9027; Sa = Staphylococcus
aureus ATCC# 6538; Bs = Bacillus subtilis ATCC# 6633; Ca = Candida
albicans ATCC# 10231; Ab = Aspergillus brasiliensis ATCC# 16404;
BTGN = Bile Tolerant Gram Negative bacteria; Cs = Clostridium
species; TSA = Trypticase Soy Agar; SDA = Sabouraud Dextrose
Agar.
[0355] ONC201 dihydrochloride when tested at the 1:300 dilution
with TSB Mod, did not meet the requirements of the USP
<61>/<62>
[0356] Microbial Limit Suitability Test. Inhibition was observed
for Staphylococcus aureus for USP <61>/<62>. Therefore,
it can be assumed that the failure to isolate the inoculated
microorganism is attributable to the bactericidal activity of
ONC201 dihydrochloride and thus it is not likely to be contaminated
with the inhibited species of microorganism.
[0357] Next, the Minimal Inhibitory Concentration (MIC) for six
imipridones was determined against wild type and
methicillin-resistant Staphylococcus aureus.
Materials and Methods
Compounds
[0358] ONC201 and ONC206 were previously solubilized at 40 mM in
DMSO. ONC212, ONC207 and ONC213 were solubilized at 20 mg/mL in
DMSO and an ONC201 linear isomer (TIC-10) was solubilized at 10
mg/mL in DMSO. Methicillin and/or vancomycin were evaluated in
parallel as positive control antibiotics and were purchased from
Sigma-Aldrich and solubilized in deionized H.sub.2O at a
concentration of 10 mg/mL.
Bacteria
[0359] The bacterial strains employed in these assays were obtained
from the American Type Culture Collection (ATCC). All bacterial
strains were propagated as recommended by the ATCC. Each strain was
stored as a frozen glycerol stock at -80.degree. C. and a 10 .mu.L
loop of the frozen stock was used to inoculate each culture for
these assays. The strains with their classification and properties
are listed in Table 8 below.
TABLE-US-00010 TABLE 8 STRAINS OF STAPHYLOCOCCUS AUREUS AND
CHARACTERISTICS ATCC # Classification Properties Assay Media 29213
Gram QC Wild Trypticase Soy Positive Type Strain Broth (TSB) 33591
Cocci Hospital Acquired Nutrient Broth Methicillin Resistant 700699
Hospital Brain Heart Acquired, MDR, Infusion Broth + Reduced 0.004
g/L Susceptibility Vancomycin to Vancomycin
Minimal Inhibitory Concentration (MIC) Determination
[0360] The susceptibility of the bacterial organisms to the test
compounds was evaluated by determining the MIC of each compound
using a micro-broth dilution analysis according to the methods
recommended by the Clinical and Laboratory Standards Institute
CLSI. All microbial strains were obtained from American Type
Culture Collections (ATCC) and cultured according to the supplier's
recommendations. Evaluation of the susceptibility of each organism
against the test compounds included a positive control
antibiotic(s). For each organism, a standardized inoculum was
prepared by direct suspension of freshly plated colonies in the
appropriate media as indicated in Table 8 to an optical density 625
nm (OD.sub.625) of 0.1 (equivalent to a 0.5 McFarland standard).
The suspended inoculum was diluted to a concentration of
approximately 1.times.10.sup.6 colony forming units per milliliter
(CFU/mL) and 100 .mu.L placed into triplicate wells of a 96-well
plate containing 100 .mu.L of test compound serially diluted 2-fold
in the appropriate broth. One hundred microliters (100 .mu.L) of
the inoculum was also added to triplicate wells containing 100
.mu.L of two-fold serial dilutions of a positive control antibiotic
and to wells containing 100 .mu.L of media only. This dilution
scheme yielded final concentrations for each microbial organism
estimated to be 5.times.10.sup.5 CFU/mL. Test compound
concentrations ranged from a high-test of 100 to a low test of 0.2
.mu.M using a two-fold dilution scheme. The plates were incubated
for 24 or 48 hours (Staphylococcus aureus 700699) at 37.degree. C.
and the microbial growth at each concentration of compound was
determined by measuring the optical density at 625 nm on a
Molecular Devices SpectraMax Plus-384 plate reader and visually by
scoring the plates +/- for bacterial growth. The MIC for each
compound was determined as the lowest compound dilution that
completely inhibited microbial growth.
Results
[0361] Six (6) imipridones were evaluated for their ability to
inhibit the growth of three strains of Staphylococcus aureus.
ONC201, ONC207, and an ONC201 linear isomer (TIC-10) were inactive
against all three strains up to a concentration of 100 .mu.g/mL.
Against wild type Staphylococcus aureus (ATCC 29213) the MIC of
ONC206, ONC212 and ONC213 was 6.25 .mu.g/mL, 3.13 .mu.g/mL and 25
.mu.g/mL, respectively. Against Staphylococcus aureus (ATCC 33591)
the MIC of ONC206, ONC212 and ONC213 was 12.5 .mu.g/mL, 3.13
.mu.g/mL and 3.13 .mu.g/mL, respectively. The activity was similar
against the MDR Staphylococcus aureus (ATCC 700699) with all three
compounds having a MIC of 12.5 .mu.g/mL. Vancomycin, the positive
control compound, was active at the expected concentration and
methicillin was found to be inactive up to a concentration of 100
.mu.g/mL against the two methicillin resistant strains of bacteria.
Data are presented in Table 9.
TABLE-US-00011 TABLE 9 MIC DETERMINATION OF 6 IMIPRIDONES FOR 3
STAPHYLOCOCCUS AUREUS STRAINS Staphylococcus aureus Staphylococcus
aureus Staphylococcus aureus Compound ATCC 29213 ATCC 33591 ATCC
700699 (48 hours) (.mu.g/mL) MIC.sub.90 MIC.sub.95 MIC.sub.99
Visual MIC.sub.90 MIC.sub.95 MIC.sub.99 Visual MIC.sub.90
MIC.sub.95 MIC.sub.99 Visual ONC201 >100 >100 >100 >100
>100 >100 >100 >100 >100 >100 >100 >100
ONC206 6.25 6.25 6.25 6.25 12.5 25 >100 12.5 12.5 12.5 25 12.5
ONC207 >100 >100 >100 >100 >100 >100 >100
>100 >100 >100 >100 >100 ONC212 3.13 3.13 3.13 3.125
3.13 6.25 100 3.125 6.25 12.5 12.5 12.5 ONC213 12.5 12.5 25 25 3.13
6.25 100 3.125 6.25 12.5 12.5 12.5 TIC-10 >100 >100 >100
>100 >100 >100 >100 >100 >100 >100 >100
>100 Methicillin -- -- -- -- >100 >100 >100 >100
>100 >100 >100 >100 Vancomycin 3.13 3.13 6.25 3.125
0.39 0.39 0.78 0.391 12.5 25 25 25
Discussion
[0362] Six (6) imipridones were evaluated for activity against 3
strains of Staphylococcus aureus. ONC201, ONC207, and TIC-10 were
inactive against all three strains. ONC206, ONC212 and ONC213 had
varying activity ranging from 3.13 .mu.g/mL to 25 .mu.g/mL against
all three bacterial strains. Relative to vancomycin the activity of
these three imipridones was equivalent or 2 to 8-fold less against
strain 29213. All three of these imipridones had 10 to 30-fold less
activity compared to vancomycin against strain 33591 and the
activity for all three compounds was 2-fold higher than vancomycin
against strain 700699.
[0363] These experiments are repeated with additional impiridones
and for additional bacteria, including both Gram-positive and
Gram-negative bacteria, such as those in Table 10.
TABLE-US-00012 TABLE 10 Gram +/ Organism Condition Gram-
Enterococcus Noscomial bacteremia, wound + faecium infections,
endocarditis, UTIs Staphylococcus Bacteremia, endocarditis + aureus
Klebsiella Pneumonia, UTIs, Upper - pneumonia respiratory tract
infections Acinetobacter Infections in ICU and - baumannii burn
patients; also being seen in general hospital and nursing homes
Pseudomonas Pneumoniae, CF - aeruginosa Enterobacter UTIs,
respiratory infections - cloacae
Example 16: Case Study of ONC201 Treatment in a Subject with
Recurrent Glioblastoma
[0364] This Example provides a case study of a 22 year old female
with recurrent glioblastoma (unmethylated MGMT, H3.3 K27M mutant)
treated with 625 mg of ONC201 once every three weeks. FIG. 28 (A)
Tumor size relative to baseline (%) of total tumor burden in the
subject. One cycle is 3 weeks. (B) Contrast MRI scans at baseline,
21, 27 and 36 weeks post-ONC201 initiation of one of 2 malignant
lesions in the subject with 625 mg q3w ONC201.
[0365] It will be appreciated by one skilled in the art that
changes could be made to the exemplary embodiments shown and
described above without departing from the broad inventive concept
thereof. It is understood, therefore, that this invention is not
limited to the exemplary embodiments shown and described, but it is
intended to cover modifications within the spirit and scope of this
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 specifically set forth here, the terms "a", "an"
and "the" are not limited to one element but instead should be read
to mean "at least one."
[0366] It is to be understood that the figures and descriptions may
have been simplified to focus on elements that are relevant for a
clear understanding, while eliminating, for purposes of clarity,
other elements that those of ordinary skill in the art will
appreciate may also comprise a portion of the invention. However,
because such elements are well known in the art, and because they
do not necessarily facilitate a better understanding of the
invention, a description of such elements is not provided
herein.
[0367] Further, to the extent that a method does not rely on the
particular order of steps set forth, the particular order should
not be construed as limitation on the claims. Claims directed to a
method should not be limited to performance of the steps in the
order written, and one skilled in the art can readily appreciate
that they can be varied and still remain within the spirit and
scope of this invention.
[0368] 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 here.
Sequence CWU 1
1
11443PRTHomo sapiens 1Met Asp Pro Leu Asn Leu Ser Trp Tyr Asp Asp
Asp Leu Glu Arg Gln1 5 10 15Asn Trp Ser Arg Pro Phe Asn Gly Ser Asp
Gly Lys Ala Asp Arg Pro 20 25 30His Tyr Asn Tyr Tyr Ala Thr Leu Leu
Thr Leu Leu Ile Ala Val Ile 35 40 45Val Phe Gly Asn Val Leu Val Cys
Met Ala Val Ser Arg Glu Lys Ala 50 55 60Leu Gln Thr Thr Thr Asn Tyr
Leu Ile Val Ser Leu Ala Val Ala Asp65 70 75 80Leu Leu Val Ala Thr
Leu Val Met Pro Trp Val Val Tyr Leu Glu Val 85 90 95Val Gly Glu Trp
Lys Phe Ser Arg Ile His Cys Asp Ile Phe Val Thr 100 105 110Leu Asp
Val Met Met Cys Thr Ala Ser Ile Leu Asn Leu Cys Ala Ile 115 120
125Ser Ile Asp Arg Tyr Thr Ala Val Ala Met Pro Met Leu Tyr Asn Thr
130 135 140Arg Tyr Ser Ser Lys Arg Arg Val Thr Val Met Ile Ser Ile
Val Trp145 150 155 160Val Leu Ser Phe Thr Ile Ser Cys Pro Leu Leu
Phe Gly Leu Asn Asn 165 170 175Ala Asp Gln Asn Glu Cys Ile Ile Ala
Asn Pro Ala Phe Val Val Tyr 180 185 190Ser Ser Ile Val Ser Phe Tyr
Val Pro Phe Ile Val Thr Leu Leu Val 195 200 205Tyr Ile Lys Ile Tyr
Ile Val Leu Arg Arg Arg Arg Lys Arg Val Asn 210 215 220Thr Lys Arg
Ser Ser Arg Ala Phe Arg Ala His Leu Arg Ala Pro Leu225 230 235
240Lys Gly Asn Cys Thr His Pro Glu Asp Met Lys Leu Cys Thr Val Ile
245 250 255Met Lys Ser Asn Gly Ser Phe Pro Val Asn Arg Arg Arg Val
Glu Ala 260 265 270Ala Arg Arg Ala Gln Glu Leu Glu Met Glu Met Leu
Ser Ser Thr Ser 275 280 285Pro Pro Glu Arg Thr Arg Tyr Ser Pro Ile
Pro Pro Ser His His Gln 290 295 300Leu Thr Leu Pro Asp Pro Ser His
His Gly Leu His Ser Thr Pro Asp305 310 315 320Ser Pro Ala Lys Pro
Glu Lys Asn Gly His Ala Lys Asp His Pro Lys 325 330 335Ile Ala Lys
Ile Phe Glu Ile Gln Thr Met Pro Asn Gly Lys Thr Arg 340 345 350Thr
Ser Leu Lys Thr Met Ser Arg Arg Lys Leu Ser Gln Gln Lys Glu 355 360
365Lys Lys Ala Thr Gln Met Leu Ala Ile Val Leu Gly Val Phe Ile Ile
370 375 380Cys Trp Leu Pro Phe Phe Ile Thr His Ile Leu Asn Ile His
Cys Asp385 390 395 400Cys Asn Ile Pro Pro Val Leu Tyr Ser Ala Phe
Thr Trp Leu Gly Tyr 405 410 415Val Asn Ser Ala Val Asn Pro Ile Ile
Tyr Thr Thr Phe Asn Ile Glu 420 425 430Phe Arg Lys Ala Phe Leu Lys
Ile Leu His Cys 435 440
* * * * *