U.S. patent application number 17/551843 was filed with the patent office on 2022-07-07 for methods of using pyruvate kinase activators.
The applicant listed for this patent is Agios Pharmaceuticals, Inc.. Invention is credited to Samuel V. Agresta, Yue Chen, Marvin Barry Cohen, Lenny Dang, Charles Kung, Elizabeth A. Merica, Bruce Alan Silver, Hua Yang.
Application Number | 20220211697 17/551843 |
Document ID | / |
Family ID | |
Filed Date | 2022-07-07 |
United States Patent
Application |
20220211697 |
Kind Code |
A1 |
Agresta; Samuel V. ; et
al. |
July 7, 2022 |
METHODS OF USING PYRUVATE KINASE ACTIVATORS
Abstract
Described herein are methods for using compounds that activate
pyruvate kinase.
Inventors: |
Agresta; Samuel V.;
(Lexington, MA) ; Chen; Yue; (Quincy, MA) ;
Cohen; Marvin Barry; (Newtown, PA) ; Dang; Lenny;
(Boston, MA) ; Kung; Charles; (Arlington, MA)
; Merica; Elizabeth A.; (Boston, MA) ; Silver;
Bruce Alan; (Dunkirk, MD) ; Yang; Hua; (Acton,
MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Agios Pharmaceuticals, Inc. |
Cambridge |
MA |
US |
|
|
Appl. No.: |
17/551843 |
Filed: |
December 15, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15735036 |
Dec 8, 2017 |
11234976 |
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PCT/US2016/036893 |
Jun 10, 2016 |
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17551843 |
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62174216 |
Jun 11, 2015 |
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International
Class: |
A61K 31/496 20060101
A61K031/496; A61P 7/06 20060101 A61P007/06 |
Claims
1-40. (canceled)
41. A method for activating one or more isozymes of pyruvate kinase
in a subject in need thereof, comprising orally administering to
the subject once or twice daily a dose of about 10 mg to about 60
mg or about 60 mg to about 200 mg of (1) Compound 1 or a
pharmaceutically acceptable salt or hydrate thereof; or (2) a
pharmaceutical composition comprising Compound 1 or a
pharmaceutically acceptable salt or hydrate thereof, and a
pharmaceutically acceptable carrier, to thereby activate pyruvate
kinase in the subject, wherein Compound 1 is
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide.
42. The method of claim 41, wherein the method comprises orally
administering to the subject once or twice daily a dose of about 10
mg to about 60 mg of Compound 1.
43. The method of claim 41, wherein the method comprises orally
administering to the subject once or twice daily a dose of about 60
mg to about 200 mg of Compound 1.
44. The method of claim 41, wherein the method comprises orally
administering to the subject once or twice daily a dose of about 30
mg or about 120 mg of Compound 1.
45. The method of claim 41, wherein the method comprises orally
administering to the subject once or twice daily a dose of about 50
mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about
175 mg or about 200 mg of Compound 1.
46. The method of claim 41, wherein the method comprises orally
administering Compound 1 once daily.
47. The method of claim 41, wherein the method comprises orally
administering Compound 1 twice daily.
48. The method of claim 41, wherein the method comprises orally
administering Compound 1 at a dose of about 15 mg about every 12
hours, about 60 mg about every 12 hours, or about 120 mg about
every 12 hours.
49. The method of claim 41, wherein the method comprises orally
administering Compound 1 at a dose of about 10 mg to about 60 mg
about every 12 hours, or about 60 mg to about 200 mg about every 12
hours.
50. The method of claim 41, wherein the method comprises orally
administering Compound 1 at a dose of about 10 mg to about 60 mg
about every 12 hours, or about 60 mg to about 200 mg about every 12
hours.
51. The method of claim 41, wherein the method comprises orally
administering Compound 1 at a dose of about 60 mg to about 200 mg
about every 24 hours.
52. The method of claim 41, wherein the method comprises orally
administering Compound 1 at a dose of about 90 mg about every 24
hours, about 120 mg about every 24 hours, about 150 mg about every
24 hours, about 180 mg about every 24 hours, or about 200 mg about
every 24 hours.
53. The method of claim 41, wherein the one or more isozymes of
pyruvate kinase is selected from PKR, PKM2, and PKL.
54. A method for activating one or more isozymes of pyruvate kinase
in a subject in need thereof, comprising orally administering to
the subject once or twice daily a dose of about 50 mg to about 300
mg of (1) Compound 1 or a pharmaceutically acceptable salt or
hydrate thereof; or (2) a pharmaceutical composition comprising
Compound 1 or a pharmaceutically acceptable salt or hydrate
thereof, and a pharmaceutically acceptable carrier, to thereby
activate pyruvate kinase in the subject, wherein Compound 1 is
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide.
55. The method of claim 54, wherein the method comprises orally
administering to the subject once or twice daily a dose of about
225 mg, about 250 mg, about 275 mg, or about 300 mg of Compound
1.
56. The method of claim 54, wherein the method comprises orally
administering Compound 1 twice daily.
57. The method of claim 54, wherein the method comprises orally
administering twice daily a dose of about 50 mg of Compound 1.
58. The method of claim 54, wherein the method comprises orally
administering twice daily a dose of about 100 mg of Compound 1.
59. The method of claim 54, wherein the one or more isozymes of
pyruvate kinase is selected from PKR, PKM2, and PKL.
Description
CLAIM OF PRIORITY
[0001] This application is a divisional of U.S. patent application
Ser. No. 15/735,036, filed Dec. 8, 2017, which is a national stage
application under 35 U.S.C. 371 of International Application No.
PCT/US2016/036893 filed Jun. 10, 2016, which claims priority from
U.S. Ser. No. 62/174,216 filed Jun. 11, 2015, which is incorporated
herein by reference in its entirety.
BACKGROUND
[0002] Pyruvate kinase deficiency (PKD) is one of the most common
enzyme defects in erythrocytes in humans due to autosomal recessive
mutations of the PKLR gene (Zanella, A., et al., Br J Haematol
2005, 130 (1), 11-25). It is also the most frequent enzyme mutation
in the central glycolytic pathway and only second to glucose-6
phosphate dehydrogenase (G6PD) deficiency (Kedar, P., et al., Clin
Genet 2009, 75 (2), 157-62) of the hexose monophosphate shunt.
[0003] Human erythrocytes are unique in that they anucleate when
mature. Immature erythocytes have nuclei but during early
erythropoiesis prior to becoming circulating reticulocytes they
extrude nuclei as well as other organelles such as mitochondria,
endoplasmic reticulum, and Golgi apparatus, in order to make room
for oxygen-carrying hemoglobin. As a result of lacking
mitochondria, mature red blood cells do not utilize any of the
oxygen they transport to economically synthesize adenosine
triphosphate (ATP) as other normal differentiated cells do.
Instead, red blood cells depend entirely on anaerobic glycolysis to
cycle nicotinamide adenine dinucleotide (NAD+) and to make ATP, an
essential energy source largely used to drive ATPase-dependent
K+/Na+ and Ca2+ pumps, in order to maintain cell membrane integrity
and pliability as they navigate through blood vessels. In PKD
disorder, two major distinctive metabolic abnormalities are ATP
depletion and concomitant increase of 2,3-diphosphoglycerate
consistent with accumulation of upper glycolytic intermediates.
Moreover, one of the consequences of decreased ATP and pyruvate
levels is lowered lactate level leading to inability to regenerate
NAD+ through lactate dehydrogenase for further use in glycolysis.
The lack of ATP disturbs the cation gradient across the red cell
membrane, causing the loss of potassium and water, which causes
cell dehydration, contraction, and crenation, and leads to
premature destruction and diminished lifetime of the red blood
cells (RBCs). Such defective RBCs are destroyed in the spleen, and
excessive hemolysis rate in the spleen leads to the manifestation
of hemolytic anemia. The exact mechanism by which PKD sequesters
newly matured RBCs in the spleen to effectively shorten overall
half-lives of circulating RBCs is not yet clear, but recent studies
suggest that metabolic dysregulation affects not only cell survival
but also the maturation process resulting in ineffective
erythropoiesis (Aizawa, S. et al., Exp Hematol 2005, 33 (11),
1292-8).
[0004] Pyruvate kinase catalyzes the transfer of a phosphoryl group
from phosphoenolpyruvate (PEP) to ADP, yielding one molecule of
pyruvate and one molecule of ATP. The enzyme has an absolute
requirement for Mg2+ and K+ cations to drive catalysis. PK
functions as the last critical step in glycolysis because it is an
essentially irreversible reaction under physiological conditions.
In addition to its role of synthesizing one of the two ATP
molecules from the metabolism of glucose to pyruvate, pyruvate
kinase is also an important cellular metabolism regulator. It
controls the carbon flux in lower-glycolysis to provide key
metabolite intermediates to feed biosynthetic processes, such as
pentose-phosphate pathway among others, in maintaining healthy
cellular metabolism. Because of these critical functions, pyruvate
kinase is tightly controlled at both gene expression and enzymatic
allostere levels. In mammals, fully activated pyruvate kinase
exists as a tetrameric enzyme. Four different isozymes (M1, M2, L
and R) are expressed from two separate genes. Erythrocyte-specific
isozyme PKR is expressed from the PKLR gene ("L gene") located on
chromosome 1q21. This same gene also encodes the PKL isozyme, which
is predominately expressed in the liver. PKLR consists of 12 exons
with exon 1 is erythroid-specific whereas exon 2 is liver-specific.
The two other mammalian isozymes PKM1 and PKM2 are produced from
the PKM gene ("M gene") by alternative splicing events controlled
by hnRNP proteins. The PKM2 isozyme is expressed in fetal tissues
and in adult proliferating cells such as cancer cells. Both PKR and
PKM2 are in fact expressed in proerythroblasts. However, upon
erythroid differentiation and maturation, PKM2 gradually is
decreased in expression and progressively replaced by PKR in mature
erythrocytes.
[0005] Clinically, hereditary PKR deficiency disorder manifests as
non-spherocytic hemolytic anemia. The clinical severity of this
disorder ranges from no observable symptoms in fully-compensated
hemolysis to potentially fatal severe anemia requiring chronic
transfusions and/or splenectomy at early development or during
physiological stress or serious infections. Most affected
individuals, who are asymptomatic, paradoxically due to enhanced
oxygen-transfer capacity, do not require any treatment. However,
for some of the most severe cases, while extremely rare
population-wise with estimated prevalence of 51 per million
(Beutler, E. Blood 2000, 95 (11), 3585-8), there is no
disease-modifying treatment available for these patients other than
palliative care (Tavazzi, D. et al., Pediatr Ann 2008, 37 (5),
303-10). These hereditary non-spherocytic hemolytic anemia (HNSHA)
patients present a clear unmet medical need.
[0006] Heterogenous genetic mutations in PKR lead to dysregulation
of its catalytic activity. Since the initial cloning of PKR and
report of a single point mutation Thr384>Met associated with a
HNSHA patient (Kanno, H. et al., Proc Natl Acad Sci USA 1991, 88
(18), 8218-21), there are now nearly 200 different reported
mutations associated with this disease reported worldwide (Zanella,
A. et al., Br J Haematol 2005, 130 (1), 11-25; Kedar, P., et al.,
Clin Genet 2009, 75 (2), 157-62; Fermo, E. et al., Br J Haematol
2005, 129 (6), 839-46; Pissard, S. et al., Br J Haematol 2006, 133
(6), 683-9). Although these mutations represent wide range genetic
lesions that include deletional and transcriptional or
translational abnormalities, by far the most common type is
missense mutation in the coding region that one way or another
affects conserved residues within domains that are structurally
important for optimal catalytic function of PKR. The pattern of
mutation prevalence seems to be unevenly distributed toward
specific ethnic backgrounds. For instance, the most frequent codon
substitutions reported for North American and European patients
appear to be Arg486>Trp and Arg510>Gln, while mutations
Arg479>His, Arg490>Trp and Asp331>Gly were more frequently
found in Asian patients (Kedar, P., et al., Clin Genet 2009, 75
(2), 157-62).
SUMMARY OF THE INVENTION
[0007] In one aspect, the present invention provides a method of
evaluating a subject, the method comprising: administering to the
subject
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1); and acquiring a value for the level of
Compound 1, the level of 2,3-diphosphoglycerate (2,3-DPG), the
level of adenosine triphosphate (ATP), or the activity of PKR in
the subject, to thereby evaluate the subject.
[0008] In some embodiments, the value for the level of Compound 1
is acquired by analyzing the plasma concentration of Compound
1.
[0009] In some embodiments, the level of 2,3-DPG is acquired by
analyzing the blood concentration of 2,3-DPG.
[0010] In some embodiments, the level of ATP is acquired by
analyzing the blood concentration of ATP.
[0011] In some embodiments, the activity of PKR is acquired by
analyzing the blood concentration of a 13C-label in the blood. For
example, 13C-labeled glucose is administered to a subject, and
incorporated into certain glycolytic intermediates in the
blood.
[0012] In some embodiments, the analysis is performed by sample
analysis of bodily fluid, such as blood, by e.g., mass
spectroscopy, e.g. LC-MS.
[0013] In another aspect, the present invention provides a method
of evaluating a subject, the method comprising acquiring, e.g.,
directly acquiring, the value for the level of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1), the level of 2,3-DPG, the level of ATP, or the
activity of PKR in a subject that has been treated with Compound 1,
to thereby evaluate the subject. In some embodiments, acquiring
comprises receiving a sample from the subject. In some embodiments,
acquiring comprises transmitting the value to another party, e.g.,
the party that administered Compound 1.
[0014] In some embodiments, the value for the level of Compound 1
is acquired by analyzing the plasma concentration of Compound
1.
[0015] In some embodiments, the level of 2,3-DPG is acquired by
analyzing the blood concentration of 2,3-DPG.
[0016] In some embodiments, the level of ATP is acquired by
analyzing the blood concentration of ATP.
[0017] In some embodiments, the activity of PKR is acquired by
analyzing the blood concentration of 13C-label in the blood. For
example, 13C-labeled glucose is administered to a subject, and
incorporated into certain glycolytic intermediates in the
blood.
[0018] In some embodiments, the analysis is performed by sample
analysis of bodily fluid, such as blood, by e.g., mass
spectroscopy, e.g. LC-MS.
[0019] In some embodiments, the subject has been administered
Compound 1 within a preselected period of less than 7 days, less
than 6 days, less than 5 days, less than 4 days, less than 3 days,
or less than 72 hours prior to the evaluation, e.g., less than 48
hours, less than 24 hours, less than 12 hours, less than 10 hours,
less than 8 hours, less than 6 hours, less than 4 hours, less than
3 hours, less than 2 hours, less than 1.5 hours, less than 1 hour,
less than 45 minutes, less than 30 minutes, or less than 15
minutes.
[0020] In some embodiments, the subject has been administered
Compound 1, e.g., orally, a dose of about 10 mg to about 3000 mg,
e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0021] In some embodiments, the subject has been administered
Compound 1, e.g., orally, a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0022] In some embodiments, the subject has been administered,
e.g., orally, Compound 1 once or twice daily.
[0023] In some embodiments, the subject has been administered
Compound 1, e.g., orally, twice daily, e.g., about every 12 hours.
In some embodiments, Compound 1 is administered to the subject at
about 10 mg to about 1000 mg about every 12 hours, e.g., about 10
mg to about 60 mg about every 12 hours, about 60 mg to about 200 mg
about every 12 hours, about 200 mg to about 500 mg about every 12
hours, about 500 mg to about 1000 mg about every 12 hours, e.g.,
about 15 mg about every 12 hours, about 60 mg about every 12 hours,
about 120 mg about every 12 hours, about 360 mg about every 12
hours, about 700 mg about every 12 hours.
[0024] In some embodiments, the subject has been administered
Compound 1, e.g., orally, once daily, e.g., about every 24 hours.
In some embodiments, Compound 1 is administered, e.g., orally, to
the subject at about 60 mg to about 200 mg about every 24 hours,
e.g., about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0025] In some embodiments, the method comprises comparing the
level of Compound 1, the level of 2,3-DPG, or the level of ATP to a
reference standard.
[0026] In some embodiments, the activity of PKR is acquired by
analyzing the blood concentration of 13C-label in the blood. For
example, 13C-labeled glucose is administered to a subject, and
incorporated into certain glycolytic intermediates in the
blood.
[0027] In some embodiments, the value for the level of Compound 1
is acquired by analyzing the plasma concentration of Compound
1.
[0028] In some embodiments, Compound 1 is present in a detectable
amount in the subject at least 2 hours, at least 3 hours, at least
4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at
least 8 hours, at least 9 hours, or at least 10 hours after
administration to the subject.
[0029] In some embodiments, the level of 2,3-DPG is acquired by
analyzing the blood concentration of 2,3-DPG.
[0030] In some embodiments, the level of ATP is acquired by
analyzing the blood concentration of ATP.
[0031] In some embodiments, the activity of PKR is acquired by
analyzing the blood concentration of a 13C-label in the blood. For
example, 13C-labeled glucose is administered to a subject, and
incorporated into certain glycolytic intermediates in the
blood.
[0032] In some embodiments, the analysis is performed by sample
analysis of bodily fluid, such as blood, by e.g., mass
spectroscopy, e.g. LC-MS.
[0033] In some embodiments, the reference standard for the level of
Compound 1, the level of 2,3-DPG, the level of ATP, or the level of
PRK activity is the level of Compound 1, the level of 2,3-DPG, the
level of ATP, or the level of PRK activity prior to administration
of Compound 1.
[0034] In some embodiments, the value for the level of Compound 1
is acquired by analyzing the plasma concentration of Compound
1.
[0035] In some embodiments, the level of 2,3-DPG is acquired by
analyzing the blood concentration of 2,3-DPG.
[0036] In some embodiments, the level of ATP is acquired by
analyzing the blood concentration of ATP.
[0037] In some embodiments, the activity of PKR is acquired by
analyzing the blood concentration of a 13C-label in the blood. For
example, 13C-labeled glucose is administered to a subject, and
incorporated into certain glycolytic intermediates in the
blood.
[0038] In some embodiments, the analysis is performed by sample
analysis of bodily fluid, such as blood, by e.g., mass
spectroscopy, e.g. LC-MS.
[0039] In some embodiments, the plasma concentration of Compound 1
is from about 10,000 ng/mL to about 1 ng/mL, e.g., about 1000 ng/mL
to about 10 ng/mL.
[0040] In some embodiments, the blood concentration of 2,3-DPG is
reduced by at least about 15% relative to the reference standard
(e.g., from about 15% to about 60%). In some embodiments, the blood
concentration of 2,3-DPG is reduced by at least about 15%, by at
least about 20%, by at least about 25%, by at least about 30%, by
at least about 35%, by at least about 40%, by at least about 45%,
by at least about 50%, by at least about 55%, by at least about
60%.
[0041] In some embodiments, the blood concentration of 2,3-DPG is
reduced for at least about 4 hours (e.g., at least about 8 hours,
at least about 12 hours, at least about 16 hours, at least about 20
hours, at least about 24 hours, at least about 36 hours, at least
about 48 hours, at least about 72 hours or longer).
[0042] In some embodiments, the blood concentration of 2,3-DPG is
reduced by at least about 15% relative to the reference standard
(e.g., from about 15% to about 60%). In some embodiments, the blood
concentration of 2,3-DPG is reduced by at least about 15%, by at
least about 20%, by at least about 25%, by at least about 30%, by
at least about 35%, by at least about 40%, by at least about 45%,
by at least about 50%, by at least about 55%, by at least about
60%, for at least about 4 hours (e.g., at least about 8 hours, at
least about 12 hours, at least about 16 hours, at least about 20
hours, at least about 24 hours, at least about 36 hours, at least
about 48 hours, at least about 72 hours or longer).
[0043] In some embodiments, the method comprises administering an
amount of Compound 1 sufficient to provide a blood concentration of
2,3-DPG that is reduced by at least 15% relative to the reference
standard (e.g., from about 15% to about 60%). In some embodiments,
the blood concentration of 2,3-DPG is reduced by at least about
15%, by at least about 20%, by at least about 25%, by at least
about 30%, by at least about 35%, by at least about 40%, by at
least about 45%, by at least about 50%, by at least about 55%, by
at least about 60%.
[0044] In some embodiments, a single administration of Compound 1
is sufficient to provide a blood concentration of 2,3-DPG reduced
by at least 15% relative to the reference standard (e.g., from
about 15% to about 60%). In some embodiments, the blood
concentration of 2,3-DPG is reduced by at least about 15%, by at
least about 20%, by at least about 25%, by at least about 30%, by
at least about 35%, by at least about 40%, by at least about 45%,
by at least about 50%, by at least about 55%, by at least about
60%.
[0045] In another aspect, the invention provides a method of
treating a subject for a disorder, e.g., hereditary non-spherocytic
hemolytic anemia; sickle cell anemia; thalassemia, e.g.
beta-thalassemia; hereditary spherocytosis; hereditary
elliptocytosis; sbetalipoproteinemia; Bassen-Kornzweig syndrome; or
paroxysmal nocturnal hemoglobinuria, comprising administering to
the subject an amount of Compound 1 sufficient to provide a blood
concentration of 2,3-DPG reduced by at least 15% relative to the
reference standard (e.g., from about 15% to about 60%). In some
embodiments, the blood concentration of 2,3-DPG is reduced by at
least about 15%, by at least about 20%, by at least about 25%, by
at least about 30%, by at least about 35%, by at least about 40%,
by at least about 45%, by at least about 50%, by at least about
55%, by at least about 60%.
[0046] In some embodiments, the reference standard is, e.g., the
2,3-DPG level or the blood ATP level, in a diseased human, e.g., a
human having a metabolic disorder or a blood disorder, e.g., a
human diagnosed with pyruvate kinase deficiency (PKD). In some
embodiments, the reference standard is, e.g., a baseline level,
e.g., the 2,3-DPG level or the blood ATP level, in the subject
prior to administration with Compound 1.
[0047] In some embodiments, the blood concentration of 2,3-DPG is
reduced for at least about 4 hours (e.g., at least about 8 hours,
at least about 12 hours, at least about 16 hours, at least about 20
hours, at least about 24 hours, at least about 36 hours, at least
about 48 hours, at least about 72 hours or longer).
[0048] In some embodiments, the subject has been administered
Compound 1 within a preselected period of less than 7 days, less
than 6 days, less than 5 days, less than 4 days, less than 3 days,
or less than 72 hours prior to the evaluation, e.g., less than 48
hours, less than 24 hours, less than 12 hours, less than 10 hours,
less than 8 hours, less than 6 hours, less than 4 hours, less than
3 hours, less than 2 hours, less than 1.5 hours, less than 1 hour,
less than 45 minutes, less than 30 minutes, or less than 15
minutes.
[0049] In some embodiments, the subject is evaluated less than 72
hours, less than 48 hours, less than 24 hours, less than 12 hours,
less than 10 hours, less than 8 hours, less than 6 hours, less than
4 hours, less than 3 hours, less than 2 hours, less than 1.5 hours,
less than 1 hour, less than 45 minutes, less than 30 minutes, or
less than 15 minutes, after administration of Compound 1.
[0050] In some embodiments, a single administration of Compound 1
is sufficient to provide a blood concentration of 2,3-DPG reduced
by at least 15% relative to the reference standard (e.g., from
about 15% to about 60%). In some embodiments, the blood
concentration of 2,3-DPG is reduced by at least about 15%, by at
least about 20%, by at least about 25%, by at least about 30%, by
at least about 35%, by at least about 40%, by at least about 45%,
by at least about 50%, by at least about 55%, by at least about
60%. In an embodiment, the blood concentration of 2,3-DPG is
reduced for at least about 4 hours (e.g., at least about 8 hours,
at least about 12 hours, at least about 16 hours, at least about 20
hours, at least about 24 hours, at least about 36 hours, at least
about 48 hours, at least about 72 hours or longer).
[0051] In some embodiments, the subject has been administered
Compound 1, e.g., orally, a dose of about 10 mg to about 3000 mg,
e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0052] In some embodiments, the subject has been administered
Compound 1, e.g., orally, a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0053] In some embodiments, the subject has been administered,
e.g., orally, Compound 1 once or twice daily.
[0054] In some embodiments, the subject has been administered
Compound 1, e.g., orally, twice daily, e.g., about every 12 hours.
In some embodiments, Compound 1 is administered to the subject at
about 10 mg to about 1000 mg about every 12 hours, e.g., about 10
mg to about 60 mg about every 12 hours, about 60 mg to about 200 mg
about every 12 hours, about 200 mg to about 500 mg about every 12
hours, about 500 mg to about 1000 mg about every 12 hours, e.g.,
about 15 mg about every 12 hours, about 60 mg about every 12 hours,
about 120 mg about every 12 hours, about 360 mg about every 12
hours, about 700 mg about every 12 hours.
[0055] In some embodiments, the subject has been administered
Compound 1, e.g., orally, once daily, e.g., about every 24 hours.
In some embodiments, Compound 1 is administered, e.g., orally, to
the subject at about 60 mg to about 200 mg about every 24 hours,
e.g., about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0056] In another aspect, the invention provides a method of
treating a subject for a disorder, e.g., hereditary non-spherocytic
hemolytic anemia; sickle cell anemia; thalassemia, e.g.
beta-thalassemia; hereditary spherocytosis; hereditary
elliptocytosis; abetalipoproteinemia; Bassen-Kornzweig syndrome; or
paroxysmal nocturnal hemoglobinuria, the method comprising orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0057] In some embodiments, the disorder is hereditary
non-spherocytic hemolytic anemia.
[0058] In some embodiments, the disorder is sickle cell anemia.
[0059] In some embodiments, the disorder is thalassemia, e.g.,
beta-thalassemia.
[0060] In some embodiments, the disorder is hereditary
spherocytosis.
[0061] In some embodiments, the disorder is hereditary
elliptocytosis.
[0062] In some embodiments, the disorder is
abetalipoproteinemia.
[0063] In some embodiments, the disorder is Bassen-Kornzweig
syndrome.
[0064] In some embodiments, the disorder is paroxysmal nocturnal
hemoglobinuria.
[0065] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0066] In some embodiments, Compound 1 is administered once or
twice daily.
[0067] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0068] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0069] Treatment methods described herein can additionally comprise
various evaluation steps prior to and/or following treatment with
Compound 1.
[0070] In some embodiments, prior to and/or after treatment with
Compound 1, the method further comprises the step of evaluating PK
and PD parameters (e.g., plasma concentration of Compound 1,
2,3-DPG and/or ATP). This evaluation may be achieved by sample
analysis of bodily fluid, such as blood by e.g., mass spectroscopy,
e.g. LC-MS.
[0071] In another aspect, the invention provides an oral dosage
unit of Compound 1, wherein the oral dosage unit consists of about
10 mg to about 3000 mg, e.g., about 10 mg to about 60 mg, about 60
mg to about 200 mg, about 200 mg to about 500 mg, about 500 mg to
about 1200 mg, about 1200 mg to about 2000 mg, or about 2000 mg to
about 3000 mg, e.g., about 30 mg, about 120 mg, about 360 mg, about
700 mg, about 1400 mg, about 2500 mg, of Compound 1.
[0072] In some embodiments, the oral dosage unit consists of about
50 mg to about 300 mg, e.g., about 50 mg, about 75 mg, about 100
mg, about 125 mg, about 150 mg, about 175 mg, 200 mg, about 225 mg,
about 250 mg, about 275 mg, about 300 mg, of Compound 1.
[0073] In another aspect, the present invention provides a method
of evaluating a subject, the method comprising administering to the
subject
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) and acquiring information regarding the
occurance of an adverse event (AE) to thereby evaluate the
subject.
[0074] In an embodiment, the adverse event is selected from
headache, nausea, vomiting, and upper respiratory tract infection.
In an embodiment, the adverse event is nausea. In an embodiment,
the adverse event is vomiting. In an embodiment, the adverse event
is upper respiratory tract infection.
[0075] In some embodiments, the subject has been administered
Compound 1, e.g., orally, a dose of about 10 mg to about 3000 mg,
e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0076] In some embodiments, the subject has been administered
Compound 1, e.g., orally, a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0077] In some embodiments, the subject has been administered,
e.g., orally, Compound 1 once or twice daily.
[0078] In some embodiments, the subject has been administered
Compound 1, e.g., orally, twice daily, e.g., about every 12 hours.
In some embodiments, Compound 1 is administered to the subject at
about 10 mg to about 1000 mg about every 12 hours, e.g., about 10
mg to about 60 mg about every 12 hours, about 60 mg to about 200 mg
about every 12 hours, about 200 mg to about 500 mg about every 12
hours, about 500 mg to about 1000 mg about every 12 hours, e.g.,
about 15 mg about every 12 hours, about 60 mg about every 12 hours,
about 120 mg about every 12 hours, about 360 mg about every 12
hours, about 700 mg about every 12 hours.
[0079] In some embodiments, the subject has been administered
Compound 1, e.g., orally, once daily, e.g., about every 24 hours.
In some embodiments, Compound 1 is administered, e.g., orally, to
the subject at about 60 mg to about 200 mg about every 24 hours,
e.g., about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0080] The present invention further provides a method for
increasing the lifetime of red blood cells (RBCs) in need thereof
comprising contacting blood with an effective amount of (1)
Compound 1 or a pharmaceutically acceptable salt thereof; (2) a
composition comprising Compound 1 or a salt thereof and a carrier;
or (3) a pharmaceutical composition comprising Compound 1 or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0081] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0082] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0083] In some embodiments, Compound 1 is administered once or
twice daily.
[0084] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0085] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0086] The present invention further provides a method for
regulating 2,3-diphosphoglycerate levels, e.g., reducing
2,3-diphosphoglycerate levels, in blood in need thereof comprising
contacting blood with an effective amount of (1) Compound 1 or a
pharmaceutically acceptable salt thereof; (2) a composition
comprising Compound 1 or a salt thereof and a carrier; or (3) a
pharmaceutical composition comprising Compound 1 or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0087] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0088] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0089] In some embodiments, Compound 1 is administered once or
twice daily.
[0090] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0091] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0092] In another aspect, the present invention provides a method
of treating a subject, the method comprising: administering to the
subject a therapeutically effective amount of (1) Compound 1 or a
pharmaceutically acceptable salt thereof; (2) a composition
comprising Compound 1 or a salt thereof and a carrier; or (3) a
pharmaceutical composition comprising Compound 1 or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier; and acquiring a value for the level of Compound
1, the level of 2,3-diphosphoglycerate (2,3-DPG), the level of
adenosine triphosphate (ATP), or the activity of PKR in the
subject, to thereby treat the subject.
[0093] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0094] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0095] In some embodiments, Compound 1 is administered once or
twice daily.
[0096] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0097] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0098] In some embodiments, the value for the level of Compound 1
is acquired by analyzing the plasma concentration of Compound
1.
[0099] In some embodiments, the level of 2,3-DPG is acquired by
analyzing the blood concentration of 2,3-DPG.
[0100] In some embodiments, the level of ATP is acquired by
analyzing the blood concentration of ATP.
[0101] In some embodiments, the activity of PKR is acquired by
analyzing the blood concentration of a 13C-label in the blood. For
example, 13C-labeled glucose is administered to a subject, and
incorporated into certain glycolytic intermediates in the
blood.
[0102] In some embodiments, the analysis is performed by sample
analysis of bodily fluid, such as blood, by e.g., mass
spectroscopy, e.g. LC-MS.
[0103] In another aspect, the present invention provides a method
for treating pyruvate kinase deficiency (PKD) in a subject in need
thereof, comprising administering to the subject a therapeutically
effective amount of (1) Compound 1 or a pharmaceutically acceptable
salt thereof; (2) a composition comprising Compound 1 or a salt
thereof and a carrier; or (3) a pharmaceutical composition
comprising Compound 1 or a pharmaceutically acceptable salt
thereof, and a pharmaceutically acceptable carrier, to thereby
treat PKD in the subject.
[0104] In some embodiments, prior to, during, and/or after
treatment with (1) Compound 1 or a pharmaceutically acceptable salt
thereof; (2) a composition comprising Compound 1 or a salt thereof
and a carrier; or (3) a pharmaceutical composition comprising
Compound 1 or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, the method further comprises
the step of evaluating for levels of Compound 1, or for the levels
of one or more intermediate(s) in the glycolysis pathway, e.g.,
evaluating for levels of one or more of 2,3-diphosphoglycerate
(2,3-DPG), adenosine triphosphate (ATP), or another intermediate in
the glycolysis pathway.
[0105] In some embodiments, the method comprises activating one or
more isozymes of pyruvate kinase, e.g., one or more of PKR, PKM2
and/or PKL isozymes.
[0106] In some embodiments, the method comprises activating wild
type PKR isozyme and/or a mutant PKR isozyme.
[0107] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0108] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0109] In some embodiments, Compound 1 is administered once or
twice daily.
[0110] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0111] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0112] In another aspect, the present invention provides a method
of activating pyruvate kinase in a subject in need thereof,
comprising administering to the subject a therapeutically effective
amount of (1) Compound 1 or a pharmaceutically acceptable salt
thereof; (2) a composition comprising Compound 1 or a salt thereof
and a carrier; or (3) a pharmaceutical composition comprising
Compound 1 or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, to thereby activate pyruvate
kinase in the subject.
[0113] In some embodiments, the method comprises activating one or
more isozymes of pyruvate kinase, e.g., one or more of PKR, PKM2
and/or PKL isozymes.
[0114] In some embodiments, the method comprises activating wild
type PKR isozyme and/or a mutant PKR isozyme. In some embodiments,
the mutant PKR isozyme is selected from G332S, G364D, T384M, G37E,
R479H, R479K, R486W, R532W, R510Q, I90N, and R490W.
[0115] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0116] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0117] In some embodiments, Compound 1 is administered once or
twice daily.
[0118] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0119] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0120] The present invention further provides a method for treating
hereditary non-spherocytic hemolytic anemia comprising
administering to a subject in need thereof a therapeutically
effective amount of (1) Compound 1 or a pharmaceutically acceptable
salt thereof; (2) a composition comprising Compound 1 or a salt
thereof and a carrier; or (3) a pharmaceutical composition
comprising Compound 1 or a pharmaceutically acceptable salt
thereof, and a pharmaceutically acceptable carrier.
[0121] In some embodiments, prior to, during, and/or after
treatment with (1) Compound 1 or a pharmaceutically acceptable salt
thereof; (2) a composition comprising Compound 1 or a salt thereof
and a carrier; or (3) a pharmaceutical composition comprising
Compound 1 or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, the method further comprises
the step of evaluating for the level of Compound 1, or for the
levels of one or more intermediate(s) in the glycolysis pathway,
e.g., evaluating for levels of one or more of
2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), or
another intermediate in the glycolysis pathway.
[0122] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0123] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0124] In some embodiments, Compound 1 is administered once or
twice daily.
[0125] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0126] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0127] The present invention further provides a method for treating
sickle cell anemia comprising administering to a subject in need
thereof a therapeutically effective amount of (1) Compound 1 or a
pharmaceutically acceptable salt thereof; (2) a composition
comprising Compound 1 or a salt thereof and a carrier; or (3) a
pharmaceutical composition comprising Compound 1 or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0128] In some embodiments, prior to, during, and/or after
treatment with (1) Compound 1 or a pharmaceutically acceptable salt
thereof; (2) a composition comprising Compound 1 or a salt thereof
and a carrier; or (3) a pharmaceutical composition comprising
Compound 1 or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, the method further comprises
the step of evaluating for the level of Compound 1, or for the
levels of one or more intermediate(s) in the glycolysis pathway,
e.g., evaluating for levels of one or more of
2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), or
another intermediate in the glycolysis pathway.
[0129] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0130] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0131] In some embodiments, Compound 1 is administered once or
twice daily.
[0132] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0133] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours, or about 200 mg about every 24 hours.
[0134] The present invention further provides a method for treating
hemolytic anemia (e.g., chronic hemolytic anemia caused by
phosphoglycerate kinase deficiency, Blood Cells Mol Dis, 2011;
46(3):206) comprising administering to a subject in need thereof a
therapeutically effective amount of (1) Compound 1 or a
pharmaceutically acceptable salt thereof; (2) a composition
comprising Compound 1 or a salt thereof and a carrier; or (3) a
pharmaceutical composition comprising Compound 1 or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0135] In some embodiments, prior to, during, and/or after
treatment with (1) Compound 1 or a pharmaceutically acceptable salt
thereof; (2) a composition comprising Compound 1 or a salt thereof
and a carrier; or (3) a pharmaceutical composition comprising
Compound 1 or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, the method further comprises
the step of evaluating for the level of Compound 1, or for the
levels of one or more intermediate(s) in the glycolysis pathway,
e.g., evaluating for levels of one or more of
2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), or
another intermediate in the glycolysis pathway.
[0136] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0137] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0138] In some embodiments, Compound 1 is administered once or
twice daily.
[0139] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0140] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours.
[0141] The present invention further provides a method for treating
thalassemia (e.g., beta-thalassemia), hereditary spherocytosis,
hereditary elliptocytosis, abetalipoproteinemia (or
Bassen-Kornzweig syndrome), paroxysmal nocturnal hemoglobinuria,
acquired hemolytic anemia (e.g., congenital anemias (e.g.,
enzymopathies)), or anemia of chronic diseases comprising
administering to a subject in need thereof a therapeutically
effective amount of (1)
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) or a pharmaceutically acceptable salt thereof;
(2) a composition comprising Compound 1 or a salt thereof and a
carrier; or (3) a pharmaceutical composition comprising Compound 1
or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier.
[0142] In some embodiments, prior to, during, and/or after
treatment with (1) Compound 1 or a pharmaceutically acceptable salt
thereof; (2) a composition comprising Compound 1 or a salt thereof
and a carrier; or (3) a pharmaceutical composition comprising
Compound 1 or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier, the method further comprises
the step of evaluating for the level of Compound 1, or for the
levels of one or more intermediate(s) in the glycolysis pathway,
e.g., evaluating for levels of one or more of
2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), or
another intermediate in the glycolysis pathway.
[0143] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0144] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0145] In some embodiments, Compound 1 is administered once or
twice daily.
[0146] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0147] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours.
[0148] The present invention further provides a method for treating
diseases or conditions that are associated with increased
2,3-diphosphoglycerate levels (e.g., liver diseases (Am J
Gastroenterol, 1987; 82(12):1283) and Parkinson's (J. Neurol,
Neurosurg, and Psychiatry 1976, 39:952) comprising administering to
a subject in need thereof a therapeutically effective amount of (1)
Compound 1 or a pharmaceutically acceptable salt thereof; (2) a
composition comprising Compound 1 or a salt thereof and a carrier;
or (3) a pharmaceutical composition comprising Compound 1 or a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier.
[0149] In some embodiments, the method comprises orally
administering to the subject a dose of about 10 mg to about 3000
mg, e.g., about 10 mg to about 60 mg, about 60 mg to about 200 mg,
about 200 mg to about 500 mg, about 500 mg to about 1200 mg, about
1200 mg to about 2000 mg, or about 2000 mg to about 3000 mg, e.g.,
about 30 mg, about 120 mg, about 360 mg, about 700 mg, about 1400
mg, about 2500 mg, of Compound 1.
[0150] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0151] In some embodiments, Compound 1 is administered once or
twice daily.
[0152] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0153] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours.
[0154] A compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) and compositions comprising Compound 1
described herein are allosteric activators of PKR mutants and
isoforms having lower activities compared to the wild type, thus
are useful for methods of the present invention. Such mutations in
PKR can affect enzyme activity (catalytic efficiency), regulatory
properties (modulation by fructose bisphosphate (FBP)/ATP), and/or
thermostability of the enzyme. Examples of such mutations are
described in Valentini et al, JBC 2002. Some examples of the
mutants that are activated by the compounds described herein
include G332S, G364D, T384M, G37E, R479H, R479K, R486W, R532W,
R510Q, I90N, and R490W. Without being bound by theory, Compound 1
affects the activities of PKR mutants by activating FBP
non-responsive PKR mutants, restoring thermostability to mutants
with decreased stability, or restoring catalytic efficiency to
impaired mutants. Compound 1 is also an activator of wild type
PKR.
[0155] In an embodiment, to increase the lifetime of the red blood
cells,
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1), composition or pharmaceutical composition
described herein is added directly to whole blood or packed cells
extracorporeally or be provided to the subject (e.g., the patient)
directly (e.g., by i.p., i.v., i.m., oral, inhalation (aerosolized
delivery), transdermal, sublingual and other delivery routes).
Without being bound by theory, Compound 1 increases the lifetime of
the RBCs, thus counteract aging of stored blood, by impacting the
rate of release of 2,3-DPG from the blood. A decrease in the level
of 2,3-DPG concentration induces a leftward shift of the
oxygen-hemoglobin dissociation curve and shifts the allosteric
equilibribrium to the R, or oxygenated state, thus producing a
therapeutic inhibition of the intracellular polymerization that
underlies sickling by increasing oxygen affinity due to the 2,3-DPG
depletion, thereby stabilizing the more soluble oxy-hemoglobin.
Accordingly, in one embodiment, Compound 1 is useful as an
antisickling agent. In another embodiment, to regulate
2,3-diphosphoglycerate, e.g. reduce 2,3-diphosphoglycerate levels,
Compound 1 is added directly to whole blood or packed cells
extracorporeally or be provided to the subject (e.g., the patient)
directly (e.g., by i.p., i.v., i.m., oral, inhalation (aerosolized
delivery), transdermal, sublingual and other delivery routes).
BRIEF DESCRIPTION OF THE DRAWINGS
[0156] FIG. 1 depicts line graphs showing PKR activity (left), ATP
levels (center), and 2,3-DPG levels (right) in whole blood from
C57/BL6 mice treated with a single dose of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) at four dose levels (1 mpk, 10 mpk, 50 mpk, and
150 mpk). Top row: Raw data for PKR activity, ATP level, and
2,3-DPG level assesments; Center row: Percent changes of each
marker for each dose normalized to vehicle treated; Bottom row:
Pharmacokinetic/pharmacodynamic correlation between Compound 1
exposure in plasma and each marker.
[0157] FIG. 2 depicts line graphs showing PKR activity (left), ATP
levels (center), and 2,3-DPG levels (right) in whole blood from
C57/BL6 mice treated with a multiple doses (13 doses, BID) of a
compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) at four dose levels (1 mpk, 10 mpk, 50 mpk, and
150 mpk). Top row: Raw data for PKR activity, ATP level, and
2,3-DPG level assesments ; Center row: Percent changes of each
marker for each dose normalized to vehicle treated; Bottom row:
Pharmacokinetic/pharmacodynamic correlation between Compound 1
exposure in plasma and each marker.
[0158] FIG. 3A and FIG. 3B depict a schematic for the determination
of PK flux activity in mice. C57/BL6 mice are administered 13 doses
(BID) of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinolin-
e-8-sulfonamide (Compound 1, 100 mpk), and whole blood samples are
removed over time. The blood samples are immediately incubated at
37.degree. C. in the presence of [U-13C6]-glucose, and the
metabolites are extracted and quantified (FIG. 3A). The resulting
data are subjected to a kinetic flux model to determine the overall
change in carbon flow through the PKR reaction (FIG. 3B).
[0159] FIG. 4 depicts a summary of the number of subjects
experiencing adverse events (AEs) by treatment group in the SAD
study, including the safety analysis set of both fed and fasted
periods.
[0160] FIG. 5 depicts a line graph showing the mean
concentration-time profiles of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) in human plasma following a single oral dose at
30 mg, 120 mg, 360 mg, 700 mg, 1400 mg, and 2500 mg.
[0161] FIG. 6 depicts the pharmacokinetic (PK) parameter values of
a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinolin-
e-8-sulfonamide Compound 1 following a single oral dose (SAD
study).
[0162] FIG. 7 depicts a line graph showing the mean
concentration-time profiles of 2,3-DPG in human blood following a
single oral dose of placebo, 30 mg, 120 mg, 360 mg, 700 mg, 1400
mg, and 2500 mg of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinolin-
e-8-sulfonamide (Compound 1).
[0163] FIG. 8 depicts a line graph showing the mean blood
concentration-time profiles of 2,3-DPG following multiple oral
doses of placebo, 120 mg, and 360 mg of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) for cohorts 1 and 2 in the MAD study.
[0164] FIG. 9 depicts a line graph showing the mean blood
concentration-time profiles of ATP following multiple oral doses of
placebo, 120 mg, and 360 mg of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) for cohorts 1 and 2 in the MAD study.
[0165] FIG. 10A depicts a line graph showing the change from
baseline concentration-time profiles of 2,3-DPG following multiple
oral doses of placebo, 15 mg (q12 h), 60 mg (q12 h), 120 mg (q12
h), 360 mg (q12 h), 700 mg (q12 h) of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1), or a single dose of Compound 1 at 120 mg (q24
h).
[0166] FIG. 10B depicts a line graph showing the change from
baseline concentration-time profiles of ATP following multiple oral
doses of placebo, 15 mg (q12 h), 60 mg (q12 h), 120 mg (q12 h), 360
mg (q12 h), 700 mg (q12 h) of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1), or a single dose of Compound 1 at 120 mg (q24
h).
[0167] FIG. 11 is a schematic illustrating the Phase 2 study
described in the Examples herein. BID (q12 h)=twice-daily (every 12
hours); DRT =data review team; PKR=pyruvate kinase red blood cell
isoform; TBD=to be determined; w=weeks.
[0168] FIG. 12 is a schematic outlining the pyruvate kinase R (PKR)
enzymatic reaction and how several pharmacodynamic (PD) assessments
contributes to a mechanistic understanding of the action of
Compound 1.
DETAILED DESCRIPTION OF THE INVENTION
[0169] The details of construction and the arrangement of
components set forth in the following description or illustrated in
the drawings are not meant to be limiting. Embodiments can be
practiced or carried out in various ways. Also, the phraseology and
terminology used herein is for the purpose of description and
should not be regarded as limiting. The use of "including,"
"comprising," or "having," "containing", "involving", and
variations thereof herein, is meant to encompass the items listed
thereafter and equivalents thereof as well as additional items.
[0170] As used herein, the term "treat" means decrease, suppress,
attenuate, diminish, arrest, or stabilize the development or
progression of a disease/disorder (e.g.e.g., hereditary
non-spherocytic hemolytic anemia; sickle cell anemia; thalassemia,
e.g. beta-thalassemia; hereditary spherocytosis; hereditary
elliptocytosis; sbetalipoproteinemia; Bassen-Kornzweig syndrome; or
paroxysmal nocturnal hemoglobinuria), lessen the severity of the
disease/disorder (e.g., hereditary non-spherocytic hemolytic
anemia; sickle cell anemia; thalassemia, e.g. beta-thalassemia;
hereditary spherocytosis; hereditary elliptocytosis;
sbetalipoproteinemia; Bassen-Kornzweig syndrome; or paroxysmal
nocturnal hemoglobinuria) or improve the symptoms associated with
the disease/disorder (e.g., e.g., hereditary non-spherocytic
hemolytic anemia; sickle cell anemia; thalassemia, e.g.
beta-thalassemia; hereditary spherocytosis; hereditary
elliptocytosis; sbetalipoproteinemia; Bassen-Kornzweig syndrome; or
paroxysmal nocturnal hemoglobinuria).
[0171] As used herein, an amount of a compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) effective to treat a disorder, or a
"therapeutically effective amount" refers to an amount of the
compound which is effective, upon single or multiple dose
administration to a subject, in treating a cell, or in curing,
alleviating, relieving or improving a subject with a disorder
beyond that expected in the absence of such treatment.
[0172] As used herein, the dosing amount refers to the free base of
Compound 1 or a pharmaceutically acceptable salt or solvate (e.g.,
hydrate) thereof.
[0173] As used herein, the term "subject" is intended to mean
human. Exemplary human subjects include a human patient (referred
to as a patient) having a disorder, e.g., a disorder described
herein or a normal subject.
[0174] As used herein, the term "acquire" or "acquiring" as the
terms are used herein, refer to obtaining possession of a physical
entity (e.g., a sample, e.g., blood sample or blood plasma sample),
or a value, e.g., a numerical value, by "directly acquiring" or
"indirectly acquiring" the physical entity or value. "Directly
acquiring" means performing a process (e.g., an analytical method)
to obtain the physical entity or value. "Indirectly acquiring"
refers to receiving the physical entity or value from another party
or source (e.g., a third party laboratory that directly acquired
the physical entity or value). Directly acquiring a value includes
performing a process that includes a physical change in a sample or
another substance, e.g., performing an analytical process which
includes a physical change in a substance, e.g., a sample,
performing an analytical method, e.g., a method as described
herein, e.g., by sample analysis of bodily fluid, such as blood by,
e.g., mass spectroscopy, e.g. LC-MS.
Methods of Treatment
[0175] In one embodiment, provided is a method for treating or
preventing a disease, condition or disorder as described herein
(e.g., treating) comprising administering to a subject in need
thereof
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1).
[0176] In some embodiments, the disorder is selected from
hereditary non-spherocytic hemolytic anemia; sickle cell anemia;
thalassemia, e.g. beta-thalassemia; hereditary spherocytosis;
hereditary elliptocytosis; sbetalipoproteinemia; Bassen-Kornzweig
syndrome; or paroxysmal nocturnal hemoglobinuria.
[0177]
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-
-sulfonamide (Compound 1) and compositions described herein can be
administered to cells in culture, e.g. in vitro or ex vivo, or to a
subject, e.g., in vivo, to treat, prevent, and/or diagnose a
variety of disorders, including those described herein below.
Compositions and Routes of Administration
[0178] The compositions delineated herein include the compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1), as well as additional therapeutic agents if
present, in amounts effective for achieving a modulation of disease
or disease symptoms, including those described herein.
[0179] The term "pharmaceutically acceptable carrier or adjuvant"
refers to a carrier or adjuvant that may be administered to a
patient, together with Compound 1, and which does not destroy the
pharmacological activity thereof and is nontoxic when administered
in doses sufficient to deliver a therapeutic amount of the
compound.
[0180] Pharmaceutically acceptable carriers, adjuvants and vehicles
that may be used in the pharmaceutical compositions provided
herewith include, but are not limited to, ion exchangers, alumina,
aluminum stearate, lecithin, self-emulsifying drug delivery systems
(SEDDS) such as d-.alpha.-tocopherol polyethyleneglycol 1000
succinate, surfactants used in pharmaceutical dosage forms such as
Tweens or other similar polymeric delivery matrices, serum
proteins, such as human serum albumin, buffer substances such as
phosphates, glycine, sorbic acid, potassium sorbate, partial
glyceride mixtures of saturated vegetable fatty acids, water, salts
or electrolytes, such as protamine sulfate, disodium hydrogen
phosphate, potassium hydrogen phosphate, sodium chloride, zinc
salts, colloidal silica, magnesium trisilicate, polyvinyl
pyrrolidone, cellulose-based substances, polyethylene glycol,
sodium carboxymethylcellulose, polyacrylates, waxes,
polyethylene-polyoxypropylene-block polymers, polyethylene glycol
and wool fat. Cyclodextrins such as .alpha.-, .beta.-, and
.gamma.-cyclodextrin, or chemically modified derivatives such as
hydroxyalkylcyclodextrins, including 2- and
3-hydroxypropyl-.beta.-cyclodextrins, or other solubilized
derivatives may also be advantageously used to enhance delivery of
compounds of the formulae described herein.
[0181] The pharmaceutical compositions provided herewith may be
orally administered in any orally acceptable dosage form including,
but not limited to, capsules, tablets, emulsions and aqueous
suspensions, dispersions and solutions. In the case of tablets for
oral use, carriers which are commonly used include lactose and corn
starch. Lubricating agents, such as magnesium stearate, are also
typically added. For oral administration in a capsule form, useful
diluents include lactose and dried corn starch. When aqueous
suspensions and/or emulsions are administered orally, the active
ingredient may be suspended or dissolved in an oily phase is
combined with emulsifying and/or suspending agents. If desired,
certain sweetening and/or flavoring and/or coloring agents may be
added.
[0182] In some embodiments, Compound 1 is orally administering to
the subject at a dose of about 10 mg to about 3000 mg, e.g., about
10 mg to about 60 mg, about 60 mg to about 200 mg, about 200 mg to
about 500 mg, about 500 mg to about 1200 mg, about 1200 mg to about
2000 mg, or about 2000 mg to about 3000 mg, e.g., about 30 mg,
about 120 mg, about 360 mg, about 700 mg, about 1400 mg, about 2500
mg.
[0183] In some embodiments, the method comprises administering,
e.g., orally, to the subject a dose of about 50 mg to about 300 mg,
e.g., about 50 mg, about 75 mg, about 100 mg, about 125 mg, about
150 mg, about 175 mg, 200 mg, about 225 mg, about 250 mg, about 275
mg, about 300 mg, of Compound 1.
[0184] In some embodiments, Compound 1 is administered once or
twice daily.
[0185] In some embodiments, Compound 1 is administered, e.g.,
orally, twice daily, e.g., about every 12 hours. In some
embodiments, Compound 1 is administered to the subject at about 10
mg to about 1000 mg about every 12 hours, e.g., about 10 mg to
about 60 mg about every 12 hours, about 60 mg to about 200 mg about
every 12 hours, about 200 mg to about 500 mg about every 12 hours,
about 500 mg to about 1000 mg about every 12 hours, e.g., about 15
mg about every 12 hours, about 60 mg about every 12 hours, about
120 mg about every 12 hours, about 360 mg about every 12 hours,
about 700 mg about every 12 hours.
[0186] In some embodiments, Compound 1 is administered, e.g.,
orally, once daily, e.g., about every 24 hours. In some
embodiments, Compound 1 is administered, e.g., orally, to the
subject at about 60 mg to about 200 mg about every 24 hours, e.g.,
about 90 mg about every 24 hours, about 120 mg about every 24
hours, about 150 mg about every 24 hours, about 180 mg about every
24 hours.
[0187] When the compositions provided herewith comprise a
combination of Compound 1 and one or more additional therapeutic or
prophylactic agents, both the compound and the additional agent
should be present at dosage levels of between about 1 to 100%, and
more preferably between about 5 to 95% of the dosage normally
administered in a monotherapy regimen. The additional agents may be
administered separately, as part of a multiple dose regimen, from
the compounds provided herewith. Alternatively, those agents may be
part of a single dosage form, mixed together with Compound 1 in a
single composition.
[0188] Lower or higher doses than those recited above may be
required. Specific dosage and treatment regimens for any particular
patient will depend upon a variety of factors, including the
activity of the specific compound employed, the age, body weight,
general health status, sex, diet, time of administration, rate of
excretion, drug combination, the severity and course of the
disease, condition or symptoms, the patient's disposition to the
disease, condition or symptoms, and the judgment of the treating
physician.
[0189] Upon improvement of a patient's condition, a maintenance
dose of a compound, composition or combination provided herewith
may be administered, if necessary. Subsequently, the dosage or
frequency of administration, or both, may be reduced, as a function
of the symptoms, to a level at which the improved condition is
retained when the symptoms have been alleviated to the desired
level. Patients may, however, require intermittent treatment on a
long-term basis upon any recurrence of disease symptoms.
Patient Selection and Monitoring
[0190] The compound
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1) can activate wild type PKR and/or mutant PKRs.
Some examples of the mutants that are activated by the compounds
described herein include G332S, G364D, T384M, G37E, R479H, R479K,
R486W, R532W, R510Q, I90N, and R490W. Accordingly, a patient and/or
subject can be selected for treatment using Compound 1 by first
evaluating the patient and/or subject to determine whether the
subject carries a mutation in PKR (for examples, one of the
mutations as described herein), and if the subject is determined to
be carrying a mutation in PKR thus is in need of activation of the
activity of the mutant PKR, then optionally administering to the
subject Compound 1. A subject can be evaluated as carrying a
mutation in PKR using methods known in the art. The subject can
also be monitored, for example, subsequent to administration of
Compound 1. In embodiments, the subject can be monitored for
evaluation of certain PK/PD parameters of Compound 1 such as levels
of Compound 1, levels of 2,3-DPG, or levels of ATP.
EXAMPLES
Example 1
Synthesis of
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfo-
namide (Compound 1)
[0191] The synthesis of Compound 1 was carried out following the
procedure described in U.S. Pat. No. 8,785,450, which is
incorporated herein by reference in its entirety.
Example 2
N-(4-(4-(cyclopropylmethyl)piperazine-1-carbonyl)phenyl)quinoline-8-sulfon-
amide (Compound 1) Activation of Pyruvate Kinase In Vivo Enhances
Red Cell Glycolysis in Mice
[0192] Pyruvate kinase deficiency (PKD) is an autosomal recessive
enzymopathy that is the most common cause of hereditary
nonspherocytic hemolytic anemia (HNSHA). PKD is a rare disease
characterized by a life-long chronic hemolysis with severe
co-morbidities. It is hypothesized that insufficient energy
production to maintain red cell membrane homeostasis promotes the
chronic hemolysis. Treatment is generally palliative, focusing on
the resultant anemia, and there are no approved drugs that directly
target mutated pyruvate kinase.
[0193] Compound 1 is an allosteric activator of the red cell
isoform of pyruvate kinase (PKR) that has recently entered Phase I
clinical trials in normal healthy volunteers. Compound 1 increases
the catalytic efficiency and enhances the protein stability of a
spectrum of recombinantly expressed PKR mutant proteins that have
been associated with PKD. PKD red cells are characterized by
changes in metabolism associated with defective glycolysis,
including a build-up of the upstream glycolytic intermediate
2,3-DPG and deficiency in the PKR product adenosine triphosphate
(ATP). PKR flux, e.g. the rate of carbon flow through the PKR
enzyme reaction, was examined in PKD patient or WT donor blood
samples by incubation of whole blood with a stable isotope tracer,
[U-13C6]-glucose. At various time points after the addition of
[U-13C6]-glucose, metabolism was quenched and metabolites were
extracted. Metabolite pool sizes and 13C label incorporation into
glycolytic intermediates were monitored by LC/MS. The rate of label
incorporation was found to be significantly slower in PKD patient
red cells, consistent with decreased glycolytic activity. Treatment
of PKD red cells with Compound 1 ex-vivo induces changes in
metabolism consistent with increased glycolytic activity including
reduced 2,3-DPG levels, increased ATP levels, and increased PKR
enzyme activity levels.
[0194] The effect of Compound 1 on red cell metabolism in vivo was
evaluated in mice. C57/BL6 mice were dosed by oral gavage either
with a single dose, or with multiple doses (13 doses, BID) of
Compound 1 for 7 days. Dose levels tested were 1 mpk, 10 mpk, 50
mpk, and 150 mpk. Following the last dose, mice were bled to
evaluate drug exposure and pharmacodynamic markers including
2,3-DPG and ATP levels, and PKR activity. Compound 1 was
demonstrated to be a well-behaved compound, with dose-proportional
increase in exposure, both in the single dose and multiple dose
studies. A single dose of Compound 1 resulted in a dose-dependent
increase in PKR activity levels (FIG. 1, left), concomitant with
reduction in 2,3-DPG levels (FIG. 1, right). There were no
significant changes in ATP levels after a single administration of
Compound 1 (FIG. 1, center). In the multiple dose studies, similar
changes in PKR activity (FIG. 2, left) and 2,3-DPG levels were
observed (FIG. 2, right). In contrast to the single-dose study, ATP
levels in the multiple dose study were observed to be robustly
increased in a dose-dependent manner (FIG. 2, center). The
resulting pharmacokinetic/pharmacodynamic correlations between
Compound 1 exposure in plasma and each pharmacodynamic marker (PKR
activity as well as ATP and 2,3-DPG levels) for both the single
dose and multiple dose studies further highlights these
observations (FIGS. 1 and 2, lower panels).
[0195] The effect of Compound 1 on PKR flux was assessed in whole
blood from mice treated with Compound 1. C57BL/6 mice were dosed by
oral gavage with Compound 1 at 100 mpk BID for 13 total doses.
Whole blood was incubated with [U-13C6]-glucose at 37.degree. C.
and the metabolite pool sizes and rate of 13C label incorporation
into glycolytic intermediates were assessed. The data were
subsequently analyzed using a mathematical kinetic flux model to
quantify the overall change in carbon flow through the PKR
reaction. Using this model, it was determined that Compound 1
treatment significantly increased glycolytic flux through the PKR
reaction as depicted schematically in FIG. 3.
[0196] Collectively, these data demonstrate that Compound 1 not
only potently binds to and activates the PKR enzyme in vivo, but
this enzyme activation induces enhanced glycolytic pathway activity
in red cells that results in profound changes in cellular
metabolism, as reflected in dramatically increased ATP levels and
reduced 2,3-DPG levels. As Compound 1 has similar potency against
the WT PKR enzyme as against tested mutant PKR enzymes in vitro,
these data support the hypothesis that Compound 1 treatment may
similarly enhance glycolytic activity in PKD patients and thus
correct the underlying pathology of PKD.
Example 3
Clinical Studies of the Safety, Tolerability, Pharmacokinetics (PK)
and Pharmacodynamics (PD) of a Pyruvate Kinase-R Activator in
Healthy Subjects
[0197] Compound 1 is a novel, first-in-class, small molecule
allosteric activator of PK-R that directly targets the underlying
metabolic defect in PKD. Pre-clinical studies demonstrated that
Compound 1 increases the activity of both wild type and various
mutated PK-R enzymes. The key objective of these first-in-human,
Phase I, randomized, double-blind, placebo-controlled single and
multiple ascending dose studies (SAD and MAD) are to identify a
safe and pharmacodynamically active dose and schedule for Compound
1 to be used in subsequent clinical studies in subjects with
pyruvate kinase deficiency.
Methods
[0198] In the single ascending dose (SAD) study, healthy men and
women (non-childbearing potential) aged 18-60 years were randomized
to receive a single oral dose of Compound 1 or placebo (P). Key
exclusion criteria included glucose 6-phosphate dehydrogenase
deficiency, blood donation, blood loss of greater than 500 mL, or
transfusion of blood or plasma within three months of screening.
Six cohorts were evaluated, each containing 8 subjects (6 subjects
receiving Compound 1, 2 subjects receiving placebo (P)), starting
with 30 mg in cohort 1 followed by 120 mg, 360 mg, 700 mg, 1400 mg
and 2500 mg in cohorts 2-6, respectively.
[0199] In the multiple ascending dose (MAD) study, 2 cohorts (120
mg BID and 360 mg BID) of 8 subjects each (6 subjects receiving
Compound 1, 2 subjects receiving placebo (P)) have completed 14
days of dosing and 2 weeks of follow-up. In both studies, safety
assessments included adverse events (AEs), vital signs, ECG and
clinical laboratory parameters. Serial blood samples were drawn for
assessment of PK and PD parameters (2,3-DPG and ATP) pre-dose and
at regular intervals thereafter at multiple doses in both the SAD
and MAD studies. Specifically, plasma concentrations of Compound 1
and blood concentrations of 2,3-DPG and ATP were analyzed by tandem
mass spectrometry methods.
[0200] The MAD study was completed by assessing safety,
tolerability, and pharmacokinetics/pharmacodynamics (PK/PD) of
Compound 1 in healthy volunteers and to identify a dosing schedule
for future trials in patients with PK deficiency. A phase 1,
single-center, randomized, double-blind, placebo-controlled MAD
study (ClinicalTrials.gov NCT02149966) was conducted in healthy men
and women (18-60 years), in 6 sequential cohorts (each cohort: n=6
Compound 1, n=2 placebo). Subjects received twice daily oral doses
of Compound 1 at 15 mg to 700 mg (q12 h), or 120 mg once daily (q24
hr) for 14 days with follow-up to Day 29. Adverse events (AEs),
laboratory parameters, ECGs, and vital signs were monitored. Plasma
concentrations of Compound 1 and whole blood 2,3-DPG and ATP levels
were measured in serial blood samples for PK/PD assessment. Hormone
levels were monitored due to pre-clinical data suggesting potential
modulation.
Results
[0201] Single Ascending Dose (SAD) Study
[0202] In the SAD, all 48 subjects enrolled completed the study,
which included 47 males and 1 female. These subjects represented a
diverse racial and ethnic pool (15 White, 31 Black, 1 Asian, 1
Native Hawaiin or other Pacific Islander, wherein 7 subjects
identify as Hispanic or Latino) and the mean age was about 40
years. Analysis of safety data indicated that 19/48 (39%) subjects
receiving Compound 1 or placebo (P) under fasted and/or fed
conditions experienced at least 1 treatment emergent adverse event
(AE) during the study (FIG. 4). All AEs were mild or moderate
(Grade 1 and 2) in severity, and the most common were nausea (n=5;
10%) and headache (n=8; 17%). In the 2 completed MAD cohorts (13
males; 3 females; mean age 44 years) 8/16 (50%) of subjects
receiving Compound 1 or placebo experienced 11 AEs. All AEs were
mild (n=10) or moderate (n=1) and the most frequent were
venipuncture bruises. There were no serious AEs, discontinuations
due to AEs, or dose-limiting toxicities in either study. Maximum
tolerated dose was not reached in the SAD and dose escalation
continues in the MAD. In SAD cohorts 1-6, exposure to single doses
of Compound 1 increased in a dose-proportional manner (mean plasma
Cmax, AUC0-12 hr and AUC0-72 hr) (FIG. 5). Absorption was rapid,
with a median Tmax of 0.75-4.0 h. The pharmacokinetic parameter
values of Compound 1 for each SAD cohort are summarized in FIG. 6.
As expected, Compound 1 had a rapid distribution or elimination
phase during the first 12 hours following dosing, with an apparent
half-life of approximately 2-4 hours (FIG. 5). The mean apparent
terminal half-life (t1/2) ranged from 17.5-20.5 hours or 50-80
hours, when concentrations were measured for 72 or 120 hours,
respectively (FIG. 6). In addition, a dose-dependent decrease in
the concentration of the pharmacodynamic marker 2,3-DPG was
observed over 24 hours following exposure to Compound 1 (with a 48%
decrease), which returned to placebo levels after 72 hours (FIG.
7). Preliminary results indicated that food has a minimal effect on
the exposure to Compound 1.
[0203] Multiple Ascending Dose (MAD) Study
[0204] In the MAD study, the pharmacokinetic results for cohorts 1
and 2 on Day 1 were consistent with those of the SAD study.
However, the pharmacokinetic parameter values of Compound 1 were
lower on Day 14 compared with that on Day 1, suggesting that
multiple doses of Compound 1 may result in an increase in the rate
of drug metabolism. The decrease in exposure on Day 14 is
consistent with pre-clinical data suggesting that Compound 1 is a
moderate inducer of cytochrome P450 3A4 (CYP3A4), which is the
major route of the oxidative metabolism of Compound 1. Similarly,
decreases in 2,3-DPG levels were also observed after administration
of the final dose in cohorts 1 and 2 of the MAD study.
Concentrations of 2,3-DPG in the blood returned to placebo levels
between 48 and 72 hours after the last dose (FIG. 8). There were
minimal increases in blood ATP levels after a single dose of
Compound 1 in the SAD study. In contrast, there were substantial
increases in ATP levels in the blood on Days 8-14of subjects in
cohorts 1 and 2 in the MAD study, and levels remained elevated
through 72 hours after the last dose (FIG. 9).
[0205] In the 6 sequential cohorts, 48 subjects (42 males and 6
females) with a mean age 41.5 (25-60) years were enrolled. Final,
unblinded safety data showed .gtoreq.1 AE in 16 out of 36 (44%)
subjects treated with Compound 1 and 4 out of the 12 (33%) placebo
(P) subjects. Treatment related .gtoreq.1 AEs was noted in 11 out
of the 36 (31%) subjects treated with Compound 1 and 3 out of the
12 (25%) placebo subjects. All treatment-related AEs were mild or
moderate (only 1 grade 3 event) in severity and often reversible
despite continued dosing. The most frequent Compound 1 related AEs
were nausea and headache, 5 out of the 36 (14%) subjects for each
(P: 0/12 (0%) nausea; headache 1 out of the 12 (8%)).
Gastrointestinal AEs occurred in subjects treated with Compound 1
only at the highest dose, 700 mg q12 h. One Grade 3 AE occurred
(Compound 1 (700 mg q12 h), elevated liver function tests (LFTs)
which resolved after treatment discontinuation). There were four
Compound 1 discontinuations: due to AEs in 2 subjects (Grade 2 drug
eruption, 60 mg q12 h; Grade 3 elevated LFTs, 700 mg q12 h), and 2
subjects withdrew consent (both had Grade 1/2 nausea and Grade 1/1
vomiting, both at 700 mg q12 h).
[0206] The highest well-tolerated dose was 360 mg q12 h (doses
between 360 and 700 were not explored). Compound 1 plasma exposure
was dose dependent with low to moderate variability in the PK
parameters of Compound 1 and its metabolite. There was a
dose-dependent decrease in 2,3-DPG and increase in ATP with the
effects plateauing at 360 mg q12 h. Decrease in 2,3-DPG was robust
after Dose 1, while the increase in ATP occurred gradually and was
strongly evident at Day 8. Change from baseline in 2,3-DPG and ATP
plateaued at .about.300 .mu.g/ml (.about.50% decrease) and
.about.175 .mu.g/ml (.about.50% increase), respectively (FIGS. 10A
and 10B, respectively). After the final Day 14 dose, 2,3-DPG
returned to levels similar to baseline between 72 and 120 hours
(FIG. 10A). ATP levels remained elevated through 120 hours
post-dose (FIG. 10B).
[0207] Compound 1 had a favorable safety profile and was
well-tolerated in healthy subjects based on preliminary analysis of
subjects receiving a single dose up to 2500 mg or multiple BID
doses up to 360 mg for up to 14 days. Compound 1 also demonstrated
a desirable PK profile, with rapid absorption, low PK variability
and dose-proportional exposure with PD effect as demonstrated on
2,3-DPG and ATP. There were no serious AEs, discontinuations due to
AEs, or dose-limiting toxicities in the SAD study, and so far no
serious AEs in the MAD study.
[0208] The dose-dependent changes in ATP and 2,3-DPG blood levels
seen in these studies are consistent with increased activity of the
glycolytic pathway, which represents the expected pharmacodynamic
effect of Compound 1. These data are consistent with pre-clinical
studies in mice described in Example 2. As Compound 1 has roughly
equipotent biochemical activity against wild type and mutant PKR
enzymes, the data support the hypothesis that Compound 1 may be
able to enhance glycolytic activity in red blood cells of patients
with PKD to address the underlying cause of the diseases.
[0209] As shown in FIG. 10A there was a decrease in 2,3-DPG in
blood with Compound 1. Mean 2,3-DPG blood levels generally
decreased from baseline over the 12-hour post-dose period following
the first dose of Compound 1 across the dose levels studied. The
rate of decrease in 2,3-DPG levels was slower at lower doses. A
large fraction of the decrease occurred after the first dose and
the decrease reached its full extent within 7 days of dosing.
Dose-related decreases in 2,3-DPG levels were observed with
increasing doses of Compound 1 at low doses of 15 and 60 mg of
Compound 1 q12 h and reached a plateau over the 120 to 700 mg q12 h
dose levels, with minimal additional decreases with higher doses.
The maximum decrease in 2,3-DPG levels was approximately 300
.mu.g/mL, an approximately 50% decrease. The concentration of
2,3-DPG returned to baseline within 72 hours after the final dose
of Compound 1.
[0210] As shown in FIG. 10B there was an increase in ATP in blood
with Compound 1. ATP levels increased during multiple dose
administration of Compound 1. Any effect of Compound 1 on ATP
levels during the 12 hours following the first dose was minimal The
increase in ATP levels reached its full extent within 10 days of
dosing. Increases in ATP levels were observed with increasing
AG-348 doses at low doses of 15 and 60 mg AG-348 q12 h and reached
a plateau over the 120 to 700 mg dose levels, with minimal
additional increases with higher doses. The maximum increase in ATP
levels was approximately 175 .mu.g/mL, an approximately 50%
increase. The concentration of ATP remained elevated for 120 hours
after the final dose of Compound 1.
Example 4
Clinical Studies of the Safety, Efficacy, Pharmacokinetics (PK),
and Pharmacodynamics (PD) of a Pyruvate Kinase-R (PKR) Activator in
Subjects with Pyruvate Kinase Deficiency
[0211] This example describes a Phase 2, open-label, two-arm,
multicenter, randomized, dose-ranging study of Compound 1 in adult
patients with pyruvate kinase deficiency (PK deficiency). This is
the first study to be conducted in patients with PK deficiency. The
key objective of this study is to evaluate the safety and
tolerability of up to 24 weeks of Compound 1 administration in
patients with PK deficiency.
Methods
[0212] In this Phase 2, open label, two arm, multicenter,
randomized, dose-ranging study, adult (male and female) patients
with pyruvate kinase deficiency receive multiple doses of Compound
1 for up to 24 weeks. Pyruvate kinase deficiency in patients is
confirmed by red blood cell (RBC) pyruvate kinase enzymatic assay.
At Week 25, patients who safely tolerate Compound 1 and demonstrate
clinical activity of Compound 1 may be eligible to roll over to a
separate safety extension study for continued treatment. Patients
who finish treatment at the end of 24 weeks or sooner will undergo
follow-up assessment 4 weeks after the last dose of the study drug.
Patients with toxicity suspected to be related to study drug will
continue follow-up until the adverse event (AE) resolves, is
declared chronic, or the patient is lost to follow-up.
[0213] Patient Selection
[0214] Patients are screened prior to randomization and Day 1 of
the treatment period to meet certain criteria. Patients included in
the study are adults (e.g., aged 18 or older) who have a medical
history/diagnosis of pyruvate kinase deficiency and who are anemic
but non-transfusion dependent.
[0215] Randomization and Dosing
[0216] Initially, up to 25 patients are randomized on an
open-label, 1:1 basis to each of two arms, e.g., 25 patients per
arm. In Arm 1, two twice-daily (BID) doses of Compound 1 are
given-300 mg Compound 1 administered orally every 12 hours (q12 h)
(BID), e.g., with a minimum of 10 hours between doses. In Arm 2, 50
mg of Compound 1 is administered orally q12 h (BID). (See FIG. 11)
Starting with Day 1, dosing is continuous (e.g., there are no rest
periods). Compound 1 is provided as a 25 mg or 100 mg (free-base
equivalent) capsule of Compound 1. The number of capsules per dose
will vary by assigned dose group. Patients will receive multiple
oral (PO) doses of Compound 1 over a 24-week treatment period.
[0217] Patient Assessments
[0218] Safety will be monitored on an on-going basis, e.g., at
regular intervals, or ad hoc as necessary. For example, adverse
events (AEs), vital signs (VS), clinical laboratory (hematology,
clinical chemistry, coagulation, and urinalysis), and
electrocardiograms (ECGs) on enrolled patients are monitored.
Additionally, available PK/PD data and indicators of clinical
activity (e.g., changes from baseline in hemoglobin (Hb)) are
assessed.
[0219] Pharmacokinetic and Pharmacodynamic Assessments
[0220] Pharmacokinetic assessments include serial blood sampling
for determination of concentration-time profiles of Compound 1 and
are conducted, e.g., following the first dose and the morning Day
15 dose. For example, additional trough levels of Compound 1 are
obtained. Compound 1 is analyzed using assays to determine
concentrations in plasma. Pharmacokinetic parameters on Day 1 and
Day 15 are computed using standard non-compartmental methods based
on observed plasma Compound 1 concentrations.
[0221] Pharmacodynamic assessments include serial blood sampling
for determination of levels of ATP and 2,3-DPG. Serial blood
sampling for determination of levels of ATP and, 2,3-DPG is
conducted, e.g., following the first dose and the morning Day 15
dose, and additional trough levels of ATP and 2,3-DPG will be
obtained. ATP and 2,3-DPG are analyzed using assays to determine
concentrations in whole blood. Pharmacodynamic parameters on Day 1
and Day 15 are computed based on observed whole blood ATP and
2,3-DPG concentrations.
[0222] In some cases, assessments include determination of PKR
activity, PKR protein, and glycolytic flux assays. Blood samples
are evaluated for PKR activity in RBCs, and assessment of
glycolytic flux in whole blood is performed through ex-vivo
labeling with 13C-glucose. Blood is also evaluated for total PKR
protein levels. Levels of additional metabolites are also assessed
in blood samples to further elucidate the mechanism and effects of
PKR activation by Compound 1. Exemplary PD markers are shown in
FIG. 12. The PKR enzyme catalyzes the PEP-to-pyruvate reaction,
with concomitant formation of ATP. Binding of Compound 1 to the PKR
tetramer can be assessed through an ex-vivo biochemical assay of
cell lysates from Compound 1 treated patients. PKR protein levels
in whole blood are assessed through Western blotting or
quantitative ELISA (or other similar assay).
[0223] The PKR Flux assay measures the change in carbon flow from
glucose through the PKR reaction to pyruvate after Compound 1
treatment. This is distinct from the target engagement measured by
the PKR activity assay because it is conducted in intact cells and
thus a more direct and functional measure of pathway activity. The
PKR Flux assay is performed by incubating freshly drawn patient
blood at 37 degrees C. with 13C6-labeled glucose. Aliquots from the
incubation reaction are taken over time and flash frozen.
Subsequent analysis by mass spectrometry reveals the rate of label
incorporation into glycolytic intermediates including DHAP,
2,3-DPG, 3-PG, and PEP. The data are fitted by mathematical
modeling to quantitate the carbon flow through the PKR reaction. An
increase in carbon flow through the PKR reaction indicates efficacy
of Compound 1. Compound 1 target engagement and stimulation of
glycolytic pathway activity has been shown in preclinical models
and healthy volunteer clinical studies to result in accumulation of
ATP and depletion of the upstream metabolite 2,3-DPG. Therefore, an
increase in ATP levels and/or a decrease in 2,3-DPG levels
indicates efficacy of Compound 1. Levels of these metabolites can
be measured by mass spectrometry from frozen whole blood
samples.
[0224] In some cases, exposure-response analysis is performed to
evaluate the relationship of Compound 1 exposure and PD effects
with changes in indicators of clinical activity (e.g., changes in
Hb levels).
[0225] Additional or alternative data/observations other than those
listed above may be reviewed. Based on the reviews, one or more of
the following steps may be implemented: [0226] Add 1 new dose arm
(Arm 3) to enroll up to 25 patients at a dose to be determined; the
dose for Arm 3 may be lower or higher than Arm 1 and Arm 2 doses,
but will not exceed 360 mg q12 h; and the dose regimen may be less
frequent than q12 h.
[0227] Having thus described several aspects of several
embodiments, it is to be appreciated various alterations,
modifications, and improvements will readily occur to those skilled
in the art. Such alterations, modifications, and improvements are
intended to be part of this invention, and are intended to be
within the spirit and scope of the invention. Accordingly, the
foregoing description and drawings are by way of example only.
* * * * *