U.S. patent application number 15/402780 was filed with the patent office on 2017-09-21 for methods of therapeutic monitoring of phenylacetic acid prodrugs.
The applicant listed for this patent is Horizon Therapeutics, LLC. Invention is credited to Masoud MOKHTARANI, Bruce SCHARSCHMIDT.
Application Number | 20170266143 15/402780 |
Document ID | / |
Family ID | 49380684 |
Filed Date | 2017-09-21 |
United States Patent
Application |
20170266143 |
Kind Code |
A1 |
SCHARSCHMIDT; Bruce ; et
al. |
September 21, 2017 |
METHODS OF THERAPEUTIC MONITORING OF PHENYLACETIC ACID PRODRUGS
Abstract
The present disclosure provides methods for adjusting the dosage
of PAA prodrugs (e.g., HPN-100, PBA) based on measurement of PAA
and PAGN in plasma and calculating the PAA:PAGN ratio so as to
determine whether PAA to PAGN conversion is saturated.
Inventors: |
SCHARSCHMIDT; Bruce; (San
Francisco, CA) ; MOKHTARANI; Masoud; (Walnut Creek,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Horizon Therapeutics, LLC |
Lake Forest |
IL |
US |
|
|
Family ID: |
49380684 |
Appl. No.: |
15/402780 |
Filed: |
January 10, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13610580 |
Sep 11, 2012 |
9561197 |
|
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15402780 |
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61636256 |
Apr 20, 2012 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 1/08 20180101; A61P
25/04 20180101; A61P 1/16 20180101; G01N 33/6812 20130101; A61P
7/00 20180101; A61P 13/00 20180101; A61P 25/28 20180101; A61P 43/00
20180101; A61P 35/00 20180101; A61P 25/00 20180101; A61K 31/192
20130101; A61P 3/00 20180101; A61P 13/12 20180101; A61P 1/00
20180101; A61K 31/216 20130101 |
International
Class: |
A61K 31/192 20060101
A61K031/192; A61K 31/216 20060101 A61K031/216; G01N 33/68 20060101
G01N033/68 |
Claims
1-13. (canceled)
14. A method of treating a urea cycle disorder in a subject
experiencing symptoms of nausea, headache, somnolence, or
confusion, having a plasma ammonia level within normal limits, and
having a plasma PAA to PAGN ratio exceeding 2.5, the method
comprising: administering to said subject, a dosage of glyceryl
tri-[4-phenylbutyrate] effective to achieve a plasma PAA to PAGN
ratio below 2.5.
15. The method of claim 14, wherein the subject has moderate
hepatic impairment.
16. The method of claim 14, wherein the subject has a low body
surface area.
17. The method of claim 14, wherein the dosage of glyceryl
tri-[4-phenylbutyrate] is administered orally.
18. The method of claim 14, further comprising measuring the plasma
ammonia level of the subject.
Description
RELATED APPLICATIONS
[0001] The present application claims priority to U.S. Provisional
Application No. 61/636,256, filed April 20, 2012, the disclosure of
which is incorporated by reference herein in its entirety,
including drawings.
BACKGROUND
[0002] Nitrogen retention disorders associated with elevated
ammonia levels include urea cycle disorders (UCDs), hepatic
encephalopathy (HE), and advanced kidney disease or kidney failure,
often referred to as end-stage renal disease (ESRD).
[0003] UCDs include several inherited deficiencies of enzymes or
transporters necessary for the synthesis of urea from ammonia,
including enzymes involved in the urea cycle. The urea cycle is
depicted in FIG. 1, which also illustrates how certain
ammonia-scavenging drugs act to assist in elimination of excessive
ammonia. With reference to FIG. 1, N-acetyl glutamine synthetase
(NAGS)-derived N-acetylglutamate binds to carbamyl phosphate
synthetase (CPS), which activates CPS and results in the conversion
of ammonia and bicarbonate to carbamyl phosphate. In turn, carbamyl
phosphate reacts with ornithine to produce citrulline in a reaction
mediated by ornithine transcarbamylase (OTC). A second molecule of
waste nitrogen is incorporated into the urea cycle in the next
reaction, mediated by arginosuccinate synthetase (ASS), in which
citrulline is condensed with aspartic acid to form argininosuccinic
acid. Argininosuccinic acid is cleaved by argininosuccinic lyase
(ASL) to produce arginine and fumarate. In the final reaction of
the urea cycle, arginase (ARG) cleaves arginine to produce
ornithine and urea. Of the two atoms of nitrogen incorporated into
urea, one originates from free ammonia (NH.sub.4.sup.+) and the
other from aspartate. UCD individuals born with no meaningful
residual urea synthetic capacity typically present in the first few
days of life (neonatal presentation). Individuals with residual
function typically present later in childhood or even in adulthood,
and symptoms may be precipitated by increased dietary protein or
physiological stress (e.g., intercurrent illness). For UCD
patients, lowering blood ammonia is the cornerstone of
treatment.
[0004] HE refers to a spectrum of neurologic signs and symptoms
believed to result from hyperammonemia, which frequently occur in
subjects with cirrhosis or certain other types of liver disease. HE
is a common manifestation of clinically decompensated liver disease
and most commonly results from liver cirrhosis with diverse
etiologies that include excessive alcohol use, hepatitis B or C
virus infection, autoimmune liver disease, or chronic cholestatic
disorders such as primary biliary cirrhosis. Patients with HE
typically show altered mental status ranging from subtle changes to
coma, features similar to patients with UCDs. It is believed that
an increase in blood ammonia due to dysfunctional liver in
detoxifying dietary protein is the main pathophysiology associated
with HE (Ong 2003).
[0005] ESRD results from a variety of causes including diabetes,
hypertension, and hereditary disorders. ESRD is manifested by
accumulation in the bloodstream of substances normally excreted in
the urine, including but not limited to urea and creatinine. This
accumulation in the bloodstream of substances, including toxins,
normally excreted in the urine is generally believed to result in
the clinical manifestations of ESRD, sometimes referred to also as
uremia or uremic syndrome. ESRD is ordinarily treated by dialysis
or kidney transplantation. To the extent that urea, per se,
contributes to these manifestations and that administration of a
phenylacetic (PAA) prodrug may decrease synthesis of urea (see,
e.g., Brusilow 1993) and hence lower blood urea concentration, PAA
prodrug administration may be beneficial for patients with
ESRD.
[0006] Subjects with nitrogen retention disorders whose ammonia
levels and/or symptoms are not adequately controlled by dietary
restriction of protein and/or dietary supplements are generally
treated with nitrogen scavenging agents such as sodium
phenylbutyrate (NaPBA, approved in the United States as
BUPHENYL.RTM. and in Europe as AMMONAPS.RTM.), sodium benzoate, or
a combination of sodium phenylacetate and sodium benzoate
(AMMONUL.RTM.). These are often referred to as alternate pathway
drugs because they provide the body with an alternate pathway to
urea for excretion of waste nitrogen (Brusilow 1980; Brusilow
1991). NaPBA is a PAA prodrug. Another nitrogen scavenging drug
currently in development for the treatment of nitrogen retention
disorders is glyceryl tri-[4-phenylbutyrate] (HPN-100), which is
described in U.S. Pat. No. 5,968,979. HPN-100, which is commonly
referred to as GT4P or glycerol PBA, is a prodrug of PBA and a
pre-prodrug of PAA. The difference between HPN-100 and NaPBA with
respect to metabolism is that HPN-100 is a triglyceride and
requires digestion, presumably by pancreatic lipases, to release
PBA (McGuire 2010), while NaPBA is a salt and is readily hydrolyzed
after absorption to release PBA.
[0007] HPN-100 and NaPBA share the same general mechanism of
action: PBA is converted to PAA via beta oxidation, and PAA is
conjugated enzymatically with glutamine to form
phenylacetylglutamine (PAGN), which is excreted in the urine. The
structures of PBA, PAA, and PAGN are set forth below:
##STR00001##
[0008] The clinical benefit of NaPBA and HPN-100 with regard to
nitrogen retention disorders derives from the ability of PAGN to
effectively replace urea as a vehicle for waste nitrogen excretion
and/or to reduce the need for urea synthesis (Brusilow 1991;
Brusilow 1993). Because each glutamine contains two molecules of
nitrogen, the body rids itself of two waste nitrogen atoms for
every molecule of PAGN excreted in the urine. Therefore, two
equivalents of nitrogen are removed for each mole of PAA converted
to PAGN. PAGN represents the predominant terminal metabolite, and
one that is stoichiometrically related to waste nitrogen removal, a
measure of efficacy in the case of nitrogen retention states.
[0009] In addition to nitrogen retention states, PAA prodrugs may
be beneficial in a variety of other disorders for which PBA and/or
PAA are believed to modify gene expression and/or exert
post-translational effects on protein function. In the case of
maple syrup urine disease (MSUD, also known as branched-chain
ketoaciduria), for example, the apparently beneficial effect of
NaPBA in lowering plasma levels of branched chain amino acids is
reported to be mediated by PBA-induced inhibition of the kinase
that regulates activity of branched chain alpha-keto acid
dehydrogenase complex or BCKDC. BCKDC is the enzyme that normally
breaks down branched-chain amino acids and is genetically defective
in MSUD patients (Bruneti-Pieri 2011). Similarly, the putative
beneficial effects of PAA prodrugs for the treatment of cancer
(Chung 2000), neurodegenerative diseases (Ryu 2005), and sickle
cell disease (Perrine 2008) all involve alteration of gene
expression and/or post-translational effects on protein function
via PBA and/or PAA.
[0010] Numerous publications reports adverse events following
administration of PBA and/or PAA (Mokhtarani 2012), and PAA is
reported to cause reversible toxicity when present in high levels
in circulation. While many of these publications have not recorded
PAA blood levels and/or temporally correlated adverse events with
PAA levels, toxicities such as nausea, headache, emesis, fatigue,
weakness, lethargy, somnolence, dizziness, slurred speech, memory
loss, confusion, and disorientation have been shown to be
temporally associated with PAA levels ranging from 499-285 .mu.g/mL
in cancer patients receiving PAA intravenously, and these
toxicities have been shown to resolve with discontinuation of PAA
administration (Thiebault 1994; Thiebault 1995). Therefore, when
administering PAA prodrugs for treatment of nitrogen retention
disorders and other conditions, it is important to optimize dosing
so as to achieve the desired therapeutic effect while minimizing
the risk of PAA associated toxicity.
SUMMARY
[0011] Provided herein is a clinically practical approach for
utilizing and interpreting blood levels of PAA and PAGN to adjust
the dose of a PAA prodrug in order to minimize the risk of
toxicities and maximize drug effectiveness.
[0012] Provided herein in certain embodiments are methods of
treating a nitrogen retention disorder or a condition for which PAA
prodrug administration is expected to be beneficial in a subject
comprising the steps of administering a first dosage of a PAA
prodrug, measuring plasma PAA and PAGN levels, calculating a plasma
PAA:PAGN ratio, and determining whether the PAA prodrug dosage
needs to be adjusted based on whether the PAA:PAGN ratio falls
within a target range. In certain embodiments, the target range is
1 to 2.5, 1 to 2, 1 to 1.5, 1.5 to 2, or 1.5 to 2.5. In certain
embodiments, a PAA:PAGN ratio above the target range indicates that
the dosage of the PAA prodrug needs to be decreased. In other
embodiments, a PAA:PAGN ratio above the target range indicates that
the dosage may need to be decreased, with the final determination
of whether to decrease the dosage taking into account other
characteristics of the subject such as biochemical profile or
clinical characteristics such as target nitrogen excretion, actual
nitrogen excretion, symptom severity, disorder duration, age, or
overall health. In certain embodiments, a PAA:PAGN ratio below the
target range indicates that the dosage of the PAA prodrug needs to
be increased. In other embodiments, a PAA:PAGN ratio below the
target range indicates that the dosage may need to be increased,
with the final determination of whether to increase the dosage
taking into account other characteristics of the subject such as
biochemical profile or clinical characteristics such as target
nitrogen excretion, actual nitrogen excretion, symptom severity,
disorder duration, age, or overall health. In certain embodiments,
a PAA:PAGN ratio that is within the target range but within a
particular subrange (e.g., 1 to 1.5 or 2 to 2.5 where the target
range is 1 to 2.5) indicates that the dosage of the PAA prodrug
does not need to be adjusted, but that the subject needs to be
subjected to more frequent monitoring. In certain embodiments, the
methods further comprise a step of administering an adjusted second
dosage if such an adjustment is determined to be necessary based on
the PAA:PAGN ratio and, optionally, other characteristics of the
subject. In other embodiments, the methods further comprise a step
of administering a second dosage that is the same as or nearly the
same as the first dosage if no adjustment in dosage is deemed to be
necessary. In certain embodiments, the nitrogen retention disorder
is UCD, HE, or ESRD. In certain embodiments, the condition for
which PAA prodrug administration is expected to be beneficial is
cancer, a neurodegenerative diseases, a metabolic disorder, or
sickle cell disease. In certain embodiments, the PAA prodrug is
HPN-100 or NaPBA. In certain embodiments, measurement of plasma PAA
and PAGN levels takes place after the first dosage of the PAA
prodrug has had sufficient time to reach steady state, such as at
48 hours to 1 week after administration.
[0013] Provided herein in certain embodiments are methods of
treating a nitrogen retention disorder or a condition for which PAA
prodrug administration is expected to be beneficial in a subject
who has previously received a first dosage of PAA prodrug
comprising the steps of measuring plasma PAA and PAGN levels,
calculating a plasma PAA:PAGN ratio, and determining whether the
PAA prodrug dosage needs to be adjusted based on whether the
PAA:PAGN ratio falls within a target range. In certain embodiments,
the target range is 1 to 2.5, 1 to 2, 1 to 1.5, 1.5 to 2, or 1.5 to
2.5. In certain embodiments, a PAA:PAGN ratio above the target
range indicates that the dosage of the PAA prodrug needs to be
decreased. In other embodiments, a PAA:PAGN ratio above the target
range indicates that the dosage may need to be decreased, with the
final determination of whether to decrease the dosage taking into
account other characteristics of the subject such as biochemical
profile or clinical characteristics such as target nitrogen
excretion, actual nitrogen excretion, symptom severity, disorder
duration, age, or overall health. In certain embodiments, a
PAA:PAGN ratio below the target range indicates that the dosage of
the PAA prodrug needs to be increased. In other embodiments, a
PAA:PAGN ratio below the target range indicates that the dosage may
need to be increased, with the final determination of whether to
increase the dosage taking into account other characteristics of
the subject such as biochemical profile or clinical characteristics
such as target nitrogen excretion, actual nitrogen excretion,
symptom severity, disorder duration, age, or overall health. In
certain embodiments, a PAA:PAGN ratio that is within the target
range but within a particular subrange (e.g., 1 to 1.5 or 2 to 2.5
where the target range is 1 to 2.5) indicates that the dosage of
the PAA prodrug does not need to be adjusted, but that the subject
needs to be subjected to more frequent monitoring. In certain
embodiments, the methods further comprise a step of administering
an adjusted second dosage if such an adjustment is determined to be
necessary based on the PAA:PAGN ratio and, optionally, other
characteristics of the subject. In other embodiments, the methods
further comprise a step of administering a second dosage that is
the same as or nearly the same as the first dosage if no adjustment
in dosage is deemed to be necessary. In certain embodiments, the
nitrogen retention disorder is UCD, HE, or ESRD. In certain
embodiments, the condition for which PAA prodrug administration is
expected to be beneficial is cancer, a neurodegenerative diseases,
a metabolic disorder, or sickle cell disease. In certain
embodiments, measurement of plasma PAA and PAGN levels takes place
after the first dosage of the PAA prodrug has had sufficient time
to reach steady state, such as at 48 hours to 1 week after
administration.
[0014] Provided herein in certain embodiments are methods of
adjusting the dosage of a PAA prodrug to be administered to a
subject comprising the steps of administering a first dosage of a
PAA prodrug, measuring plasma PAA and PAGN levels, calculating a
plasma PAA:PAGN ratio, and determining whether the PAA prodrug
dosage needs to be adjusted based on whether the PAA:PAGN ratio
falls within a target range. In certain embodiments, the target
range is 1 to 2.5, 1 to 2, 1 to 1.5, 1.5 to 2, or 1.5 to 2.5. In
certain embodiments, a PAA:PAGN ratio above the target range
indicates that the dosage of the PAA prodrug needs to be decreased.
In other embodiments, a PAA:PAGN ratio above the target range
indicates that the dosage may need to be decreased, with the final
determination of whether to decrease the dosage taking into account
other characteristics of the subject such as biochemical profile or
clinical characteristics such as target nitrogen excretion, actual
nitrogen excretion, symptom severity, disorder duration, age, or
overall health. In certain embodiments, a PAA:PAGN ratio below the
target range indicates that the dosage of the PAA prodrug needs to
be increased. In other embodiments, a PAA:PAGN ratio below the
target range indicates that the dosage may need to be increased,
with the final determination of whether to increase the dosage
taking into account other characteristics of the subject such as
biochemical profile or clinical characteristics such as target
nitrogen excretion, actual nitrogen excretion, symptom severity,
disorder duration, age, or overall health. In certain embodiments,
a PAA:PAGN ratio that is within the target range but within a
particular subrange (e.g., 1 to 1.5 or 2 to 2.5 where the target
range is 1 to 2.5) indicates that the dosage of the PAA prodrug
does not need to be adjusted, but that the subject needs to be
subjected to more frequent monitoring. In certain embodiments, the
methods further comprise a step of administering an adjusted second
dosage if such an adjustment is determined to be necessary based on
the PAA:PAGN ratio and, optionally, other characteristics of the
subject. In other embodiments, the methods further comprise a step
of administering a second dosage that is the same as or nearly the
same as the first dosage if no adjustment in dosage is deemed to be
necessary. In certain embodiments, measurement of plasma PAA and
PAGN levels takes place after the first dosage of the PAA prodrug
has had sufficient time to reach steady state, such as at 48 hours
to 1 week after administration.
[0015] Provided herein in certain embodiments are methods of
determining whether a first dosage of a PAA prodrug can be safely
administered to a subject comprising the steps of administering the
first dosage of a PAA prodrug, measuring plasma PAA and PAGN
levels, calculating a plasma PAA:PAGN ratio, and determining
whether the first dosage can be safely administered based on
whether the PAA:PAGN ratio falls above a target range. In certain
embodiments, the target range is 1 to 2.5, 1 to 2, 1 to 1.5, 1.5 to
2, or 1.5 to 2.5. In certain embodiments, a PAA:PAGN ratio above
the target range indicates that the first dosage is unsafe and
needs to be decreased. In other embodiments, a PAA:PAGN ratio above
the target range indicates that the first dosage is potentially
unsafe and may need to be decreased, with the final determination
of whether to decrease the dosage taking into account other
characteristics of the subject such as biochemical profile or
clinical characteristics such as target nitrogen excretion, actual
nitrogen excretion, symptom severity, disorder duration, age, or
overall health. In certain embodiments, a PAA:PAGN ratio that is
within the target range but within a particular subrange (e.g., 2
to 2.5 where the target range is 1 to 2.5) indicates that the first
dosage is likely safe, but that the subject needs to be subjected
to more frequent monitoring. In certain embodiments, the methods
further comprise a step of administering an adjusted second dosage
if such an adjustment is determined to be necessary based on the
PAA:PAGN ratio and, optionally, other characteristics of the
subject. In certain embodiments, measurement of plasma PAA and PAGN
levels takes place after the first dosage of the PAA prodrug has
had sufficient time to reach steady state, such as at 48 hours to 1
week after administration.
[0016] Provided herein in certain embodiments are methods of
determining whether a first dosage of a PAA prodrug is likely to be
effective for treating a nitrogen retention disorder or another
disorder for which PAA prodrug administration is expected to be
beneficial comprising the steps of administering the first dosage
of a PAA prodrug, measuring plasma PAA and PAGN levels, calculating
a plasma PAA:PAGN ratio, and determining whether the first dosage
is likely to be effective based on whether the PAA:PAGN ratio falls
below a target range. In certain embodiments, the target range is 1
to 2.5, 1 to 2, 1 to 1.5, 1.5 to 2, or 1.5 to 2.5. In certain
embodiments, a PAA:PAGN ratio below the target range indicates that
the first dosage is unlikely to be effective needs to be increased.
In other embodiments, a PAA:PAGN ratio below the target range
indicates that the first dosage is potentially ineffective and may
need to be increased, with the final determination of whether to
increase the dosage taking into account other characteristics of
the subject such as biochemical profile or clinical characteristics
such as target nitrogen excretion, actual nitrogen excretion,
symptom severity, disorder duration, age, or overall health. In
certain embodiments, a PAA:PAGN ratio that is within the target
range but within a particular subrange (e.g., 1 to 1.5 where the
target range is 1 to 2.5) indicates that the first dosage is likely
effective, but that the subject needs to be subjected to more
frequent monitoring. In certain embodiments, the methods further
comprise a step of administering an adjusted second dosage if such
an adjustment is determined to be necessary based on the PAA:PAGN
ratio and, optionally, other characteristics of the subject. In
certain embodiments, measurement of plasma PAA and PAGN levels
takes place after the first dosage of the PAA prodrug has had
sufficient time to reach steady state, such as at 48 hours to 1
week after administration.
[0017] In certain embodiments, methods are provided for optimizing
the therapeutic efficacy of a PAA prodrug in a subject who has
previously been adminsitered a first dosage of PAA prodrug
comprising the steps of measuring plasma PAA and PAGN levels,
calculating a plasma PAA:PAGN ratio, and determining whether the
PAA prodrug dosage needs to be adjusted based on whether the
PAA:PAGN ratio falls within a target range. In certain embodiments,
the target range is 1 to 2.5, 1 to 2, 1 to 1.5, 1.5 to 2, or 1.5 to
2.5. In certain embodiments, a PAA:PAGN ratio above the target
range indicates that the dosage of the PAA prodrug needs to be
decreased. In other embodiments, a PAA:PAGN ratio above the target
range indicates that the dosage may need to be decreased, with the
final determination of whether to decrease the dosage taking into
account other characteristics of the subject such as biochemical
profile or clinical characteristics such as target nitrogen
excretion, actual nitrogen excretion, symptom severity, disorder
duration, age, or overall health. In certain embodiments, a
PAA:PAGN ratio below the target range indicates that the dosage of
the PAA prodrug needs to be increased. In other embodiments, a
PAA:PAGN ratio below the target range indicates that the dosage may
need to be increased, with the final determination of whether to
increase the dosage taking into account other characteristics of
the subject such as biochemical profile or clinical characteristics
such as target nitrogen excretion, actual nitrogen excretion,
symptom severity, disorder duration, age, or overall health. In
certain embodiments, a PAA:PAGN ratio that is within the target
range but within a particular subrange (e.g., 1 to 1.5 or 2 to 2.5
where the target range is 1 to 2.5) indicates that the dosage of
the PAA prodrug does not need to be adjusted, but that the subject
needs to be subjected to more frequent monitoring. In certain
embodiments, the methods further comprise a step of administering
an adjusted second dosage if such an adjustment is determined to be
necessary based on the PAA:PAGN ratio and, optionally, other
characteristics of the subject. In other embodiments, the methods
further comprise a step of administering a second dosage that is
the same as or nearly the same as the first dosage if no adjustment
in dosage is deemed to be necessary. In certain embodiments,
measurement of plasma PAA and PAGN levels takes place after the
first dosage of the PAA prodrug has had sufficient time to reach
steady state, such as at 48 hours to 1 week after
administration.
[0018] In certain embodiments, methods are provided for obtaining a
plasma PAA:PAGN ratio within a target range in a subject comprising
the steps of administering a first dosage of a PAA prodrug,
measuring plasma PAA and PAGN levels, calculating a plasma PAA:PAGN
ratio, and determining whether the PAA:PAGN ratio falls within the
target range. If the PAA:PAGN ratio does not fall within the target
range, an adjusted second dosage is administered, and these steps
are repeated until a plasma PAA:PAGN ratio falling within the
target range is achieved. In certain embodiments, the target range
is 1 to 2.5, 1 to 2, 1 to 1.5, 1.5 to 2, or 1.5 to 2.5. In certain
embodiments, a PAA:PAGN ratio above the target range indicates that
the dosage of the PAA prodrug needs to be decreased and a PAA:PAGN
ratio below the target range indicates that the dosage of the PAA
prodrug needs to be increased. In certain embodiments, measurement
of plasma PAA and PAGN levels takes place after the first dosage of
the PAA prodrug has had sufficient time to reach steady state, such
as at 48 hours to 1 week after administration.
BRIEF DESCRIPTION OF DRAWINGS
[0019] FIG. 1: Urea cycle.
[0020] FIG. 2: Plasma PAA levels versus plasma PAA:PAGN ratio in
(A) all subjects combined (healthy adults, patients age 2 months
and above with UCDs, and patients with cirrhosis), (B) patients age
2 months and above with UCDs, and (C) patients with cirrhosis.
[0021] FIG. 3: Estimated probability (95% confidence interval
(c.i.)) of correctly detecting elevated plasma PAA:PAGN ratio
(.gtoreq.2.0) with a single blood sample at a designated time.
[0022] FIG. 4: Distribution of plasma PAA:PAGN ratio (log scale) by
time since dosing (hours) and category of maximum PAA:PAGN ratio in
all subjects combined.
[0023] FIG. 5: Distribution of plasma PAA concentrations (.mu.g/mL)
by PAA:PAGN ratio for (A) all subjects and (B) UCD and HE
subjects.
DETAILED DESCRIPTION
[0024] The following description of the invention is merely
intended to illustrate various embodiments of the invention. As
such, the specific modifications discussed are not to be construed
as limitations on the scope of the invention. It will be apparent
to one skilled in the art that various equivalents, changes, and
modifications may be made without departing from the scope of the
invention, and it is understood that such equivalent embodiments
are to be included herein.
[0025] The enzymes responsible for beta oxidation of PBA to PAA are
present in most cell types capable of utilizing fatty acids as
energy substrates, and the widespread distribution of these enzymes
presumably accounts for the rapid and essentially complete
conversion of PBA to PAA. However, the enzymes that conjugate PAA
with glutamine to form PAGN are found primarily in the liver and to
a lesser extend in kidneys (Moldave 1957). Therefore, the
conversion of PAA to PAGN may be affected under several
circumstances, including the following: a) if conjugation capacity
is saturated (e.g., by high doses of PAA prodrug); b) if
conjugation capacity is compromised (e.g., by severe hepatic and/or
renal dysfunction); c) if the substrate (glutamine) for PAA to PAGN
conjugation is rate limiting; d) genetically determined variability
(i.e., polymorphisms) in the enzymes responsible for PAA to PAGN
conversion, or e) in young children, since the capacity to convert
PAA to PAGN varies with body size measured as body surface area
(Monteleone 2012). The presence of any one of these conditions may
lead to accumulation of PAA in the body, which causes reversible
toxicity.
[0026] The goal of PAA prodrug administration in subjects with
nitrogen retention disorders is to provide a sufficient dosage to
obtain a desired level of nitrogen removal while avoiding excess
build-up of PAA. The goal of PAA prodrug administration in patients
without a nitrogen retention disorder (e.g., a neurodegenerative
disease) is to achieve circulating metabolite levels necessary to
produce a clinical benefit by alteration of gene expression and/or
protein folding or function. However, there are several
difficulties associated with determining the proper dosage in
patients with nitrogen retention disorders.
[0027] Plasma PAA and PAGN levels are affected by various factors,
including timing of the blood draw in relation to drug
administration, hepatic function, availability of metabolizing
enzymes, and availability of substrates required for metabolism. A
random PAA level drawn during an outpatient visit to determine if
levels are in the toxicity range without considering concomitant
PAGN level is insufficient to inform dosing. First, PAA levels vary
many-fold over the course of the day, fluctuating a great deal
between peak and trough levels. For example, in the Hyperion
pivotal study evaluating HPN-100 for use in treating adult UCD
(Study ID HPN-100-006, Clinical Trials ID NCT00992459), serial
blood samples were obtained for PK studies over a 24 hour period
during which subjects were receiving HPN-100 or NaPBA. The
fluctuation index for PAA over a 24 hour period, which represents
the fluctuation between maximum concentration (typically observed
after the last daily dose or at approximately 12 hours) and minimum
concentration (typically observed in the morning after overnight
fasting or at 0 hours), indicated a very high degree of variability
(2150% for NaPBA and 1368% for HPN-100). Therefore, a single plasma
PAA level may not be representative of the highest PAA level a
patient may experience during the day. Second, a high plasma PAA
level may only be indicative of the high doses a subject is
receiving rather than a point of concern if the subject is
effectively conjugating PAA with glutamine to form PAGN. Therefore,
basing dose adjustment on only on a high PAA level without
considering concomitant plasma PAGN level may result in unnecessary
dose reduction and under-treatment of the patient. Conversely, a
PAA level seemingly below the levels associated with toxicity might
be taken as an indication of satisfactory dosing without
appreciating the fact that the concomitant PAGN level may not be
proportional to PAA, indicating that PAA is not being efficiently
utilized and may be accumulating.
[0028] Previous studies have shown that conversion of PAA to PAGN
is a saturable process that varies considerably among individuals
(see, e.g., Monteleone 2012), and that patients with hepatic
impairment have higher PAA levels than patients without hepatic
impairment (Ghabril et al., "Glycerol phenylbutyrate (GPD)
administration in patients with cirrhosis and episodic hepatic
encephalopathy (HE)," submitted to Digestive Disease Week, 2012).
If PAGN formation is affected by any of the above factors, PAA will
be accumulated and waste nitrogen may not be removed from the body.
Previous studies have also shown that a small proportion of
individuals, including both healthy adults ad patients with UCDs or
HE, have higher PAA levels than the remainder of the population,
presumably due to individual differences in conjugating PAA to
PAGN, and that PAA levels fluctuate many-fold during the day
depending on the dose and the timing of blood sample relative to
the last dose so that a single plasma level may not be informative
(Lee 2010; Lichter 2011).
[0029] Although the goal of PAA prodrug therapy for nitrogen
retention disorders is to achieve ammonia levels within a normal
limit, there is no correlation between plasma PAA levels and blood
ammonia. Nitrogen retention disorder subjects are normally "dosed
to effect," meaning that subjects with absent or severely deficient
urea synthetic capacity require higher doses of PAA prodrugs than
do mildly deficient UCD patients. These higher dosages are
generally associated with higher PAA levels, such that the
conventional PK/PD response (higher active moiety, i.e., PAA,
correlates with lower harmful substance, i.e., ammonia) does not
apply. Therefore, there is no single target plasma PAA level that
can be applied to patients with UCDs or other nitrogen retention
disorders based on their blood ammonia.
[0030] Patients with severe hepatic impairment are at increased
risk of PAA accumulation due to inadequate levels of PAA
conjugating enzymes if treated with PAA-prodrugs. UCD patients
without hepatic impairment whose PAA conjugating enzymes are
readily saturated are also at increased risk of PAA accumulation if
treated with PAA-producing compounds. Other patients without
nitrogen retention are at increased risk of PAA accumulation due to
limited availability of glutamine as the substrate to form PAGN if
treated with PAA-producing compounds, which accumulates in patients
with nitrogen retention states.
[0031] WO09/134460 and WO10/025303 disclose methods for determining
an effective dosage of a PAA prodrug based on urinary PAGN levels,
which was found to be a more reliable indictor of effective dosage
than plasma levels of PAA or other metabolites. Although such
measurements are highly useful for evaluating waste nitrogen
removal, they do not provide complete information regarding a
subject's ability to utilize the prodrug.
[0032] Since PAA, PAGN, and ammonia levels do not provide the
information necessary to determine whether a subject is effectively
converting PBA to PAGN (i.e., effectively utilizing the PAA
prodrug), there is a need for improved methods of adjusting PAA
prodrug dosage and incorporating such adjustments into methods of
treating nitrogen retention disorders.
[0033] As disclosed herein, plasma PAA:PAGN ratio has been found to
provide an unexpectedly accurate measure of PAA prodrug metabolism
in subjects with nitrogen retention disorders and/or hepatic
impairment. It was found that subjects who can readily convert PAA
to PAGN and have not reached the saturation point with respect to
PAA to PAGN conversion will have a plasma PAA:PAGN ratio of 2.5 or
below (when both are measured in .mu.g/mL), and that subjects with
PAA:PAGN ratios above 2.5 have a significantly higher chance of
experience a PAA level above 400 .mu.g/mL or 500 .mu.g/mL over a 24
hour period. A PAA/PAGN ratio of less than 2.5 was associated
primarily with healthy adult or adolescent subjects and normal
liver function, with subjects having a ratio below 2.5 exhibiting a
1% probability of experiencing a PAA level greater than 400
.mu.g/mL and almost no chance of exhibiting a PAA level greater
than 500 .mu.g/mL at any point during a 24 hour period. A ratio
greater than 2.5, on the other hand, was generally seen in subjects
with moderate hepatic impairment, a subset of healthy subjects or
UCD patients with relatively lower saturation point and difficulty
conjugating PAA to form PAGN, and patients with a low body surface
area. Subjects with a ratio greater than 2.5, on the other hand,
exhibited a 20-36% likelihood of experiencing a PAA level greater
than 400 .mu.g/mL during the day, and an approximately 10%
likelihood of experiencing a PAA level of 500 .mu.g/L or greater.
In subjects with a ratio greater than 3, the likelihood of
experiencing a PAA level higher than 500 .mu.g/mL increased to as
high as 25%. These results show that a plasma PAA:PAGN ratio
exceeding 2.5 in a patient with unexplained neurological adverse
events and normal ammonia indicates that dosage adjustment should
be considered. Thus, plasma PAA:PAGN ratio provides a clinically
useful surrogate for evaluating the efficiency of PAA to PAGN
conversion.
[0034] Plasma PAA:PAGN ratio indicates whether a PAA prodrug is
being effectively utilized and scavenging nitrogen, and therefore
provides an indirect and simple measure of saturation of
conjugating enzymes, availability of substrate, and possible effect
of hepatic or renal impairment on this process. Calculating this
ratio will allow effective treatment and dose adjustment in
subjects with known hepatic impairment, subjects presenting with
signs and symptoms overlapping between hyperammonemia and PAA
toxicities, and subjects who are not clinically controlled despite
increasing the dosage of drugs.
[0035] One of ordinary skill in the art would generally not
consider the ratio of an active metabolite such as PAA to a
terminal metabolite such as PAGN when making therapeutic decisions
because they would expect that higher levels of the active
metabolite would result in a proportionately higher response (as
measured by PAGN production) and increased efficacy (i.e., waste
nitrogen removal). However, the results provided herein show that
the use of plasma PAA:PAGN ratios to evaluate and adjust PAA
prodrug dosage is unexpectedly superior to the use of PAA or PAGN
levels alone. Once a subject exceeds a specific PAA:PAGN ratio,
there is a high likelihood that they are not effectively utilizing
the active moiety and that further increasing PAA prodrug dosage
may not increase efficacy and may actually result in PAA
accumulation and toxicity.
[0036] Based on these findings, methods are provided herein for
treating nitrogen retention disorders and evaluating and adjusting
the dosage of a PAA prodrug based on plasma PAA:PAGN ratio.
Generally, these methods comprise steps of measuring plasma PAA and
PAGN levels, calculating the PAA:PAGN ratio, and determining
whether the ratio falls within a target range, with this
determination being used at least in part to decide whether to
adjust PAA prodrug dosage. In these methods, PAA:PAGN ratio can be
used to ensure that urinary PAGN output, plasma ammonia
concentration, and/or PAA levels fall within a predefined target
range. Such methods represent an improvement over previously
developed methods for evaluating PAA prodrug dosage and efficacy in
that they allow for more accurate dosing, greater efficacy, and
decreased risk of toxicity associated with PAA accumulation.
[0037] Disclosed herein are target ranges for the ratio of plasma
PAA to PAGN in subjects who are receiving PAA prodrug therapy. In
certain embodiments, a subject exhibiting a PAA:PAGN ratio falling
within a target range is classified as properly dosed, meaning that
they do not require a PAA prodrug dosage adjustment, while a
subject exhibiting a PAA:PAGN ratio falling outside the target
range is classified as improperly dosed, meaning that they require
an adjustment in PAA prodrug dosage. In certain of these
embodiments, a subject exhibiting a plasma PAA:PAGN ratio falling
above a target range is classified as requiring a decreased dosage
of PAA prodrug, while a subject exhibiting a plasma PAA:PAGN ratio
falling below a target range is classified as requiring an
increased dosage of PAA prodrug. In other embodiments, a subject
exhibiting a plasma PAA:PAGN ratio falling above a target range is
classified as requiring a decreased dosage of PAA prodrug, while a
subject exhibiting a plasma PAA:PAGN ratio falling below a target
range is classified as potentially requiring an increase in PAA
prodrug dosage. In still other embodiments, a subject exhibiting a
plasma PAA:PAGN ratio falling above a target range is classified as
potentially requiring a decreased dosage of PAA prodrug, while a
subject exhibiting a plasma PAA:PAGN ratio falling below a target
range is classified as potentially requiring an increase in PAA
prodrug dosage. In those embodiments where a subject is classified
as potentially requiring an increase or decrease in PAA prodrug
dosage based on their PAA:PAGN ratio, a decision as to whether to
increase or decrease dosage may be based on one or more additional
characteristics of the subject such as biochemical profile or
clinical characteristics such as target nitrogen excretion, actual
nitrogen excretion, symptom severity, disorder duration, age, or
overall health.
[0038] In certain embodiments, the target range for plasma PAA:PAGN
ratio is 1 to 2.5, meaning that a subject exhibiting a PAA:PAGN
falling within this range is classified as properly dosed. In other
embodiments, the target range for plasma PAA:PAGN ratio is 1 to 2,
1 to 1.5, 1.5 to 2, or 1.5 to 2.5. In certain of those embodiments
where the target range is 1 to 2.5, a subject with a PAA:PAGN ratio
above 2.5 is classified as requiring a decrease in PAA prodrug
dosage, while a subject with a PAA:PAGN ratio falling below 1 is
classified as potentially requiring an increase in PAA prodrug
dosage. In certain of these embodiments, a subject is necessarily
classified as requiring an increase in PAA prodrug dosage if their
ratio is below 1. In other embodiments, a subject with a PAA:PAGN
ratio of less than 1 is only classified as requiring an increase in
PAA prodrug dosage if one or more additional clinical or
biochemical characteristics are satisfied (e.g., the subject is
exhibiting severe symptoms of a nitrogen retention disorder).
[0039] In certain embodiments, the target range for plasma PAA:PAGN
ratio may comprise one or more subranges, with subjects falling
within different subranges being treated differently despite
falling within the target range. For example, where a target range
is 1 to 2.5, a subject exhibiting a PAA:PAGN ratio below 1 or above
2.5 may be classified as requiring an adjustment in PAA prodrug
dosage. Within the target range, subjects with a PAA:PAGN ratio
falling within a particular subrange may be treated as properly
dosed, improperly dosed (i.e., requiring a dosage adjustment), or
properly dosed but requiring more frequent monitoring. For example,
subjects having a PAA:PAGN ratio greater than 2 but not greater
than 2.5 may be classified as properly dosed but requiring more
frequent monitoring.
[0040] In certain embodiments, subrange boundaries or the treatment
of subjects falling within a particular subrange will depend in
part on a subject's specific characteristics, including for example
biochemical profile or clinical characteristics such as target
nitrogen excretion, actual nitrogen excretion, symptom severity,
disorder duration, age, or overall health. For example, in certain
embodiments a first subject with a PAA:PAGN ratio falling within
the subrange of 2 to 2.5 may be classified as properly dosed but
requiring frequent monitoring, while a second subject falling
within the same subrange may be classified as requiring a decreased
dosage of PAA prodrug. Similarly, a first subject with a PAA:PAGN
ratio falling within the subrange of 1 to 1.5 may be classified as
properly dosed but requiring frequent monitoring, while a second
subject falling within the same subrange may be classified as
requiring an increased dosage of PAA prodrug. For example, a
subject who has recently exhibited particularly acute symptoms
associated with a particular disorder may be classified as
requiring an increased dosage of PAA prodrug when exhibiting a
PAA:PAGN ratio of 1 to 1.5, while a subject who is clinically
controlled may be classified as properly dosed despite a ratio
falling within the same subrange.
[0041] In certain embodiments, methods are provided herein for
treating a nitrogen retention disorder or a condition for which PAA
prodrug administration is expected to be beneficial in a subject
that has previously received a first dosage of a PAA prodrug. These
methods comprise measuring plasma PAA and PAGN levels, calculating
the plasma PAA:PAGN ratio, determining whether the PAA prodrug
dosage needs to be adjusted based on whether the PAA:PAGN ratio
falls within a target range, and administering a second dosage of
the PAA prodrug. In certain embodiments, the target range for
PAA:PAGN ratio is 1 to 2.5 or 1 to 2. In certain of these
embodiments, the second dosage is greater than the first dosage if
the PAA:PAGN ratio is less than 1 (i.e., the dosage is increased)
and less than the first dosage if the PAA:PAGN ratio is greater
than 2.5 (i.e., the dosage is decreased). In other embodiments, the
second dosage may or may not be greater than the first dosage if
the PAA:PAGN ratio is less than 1, depending on one or more other
characteristics of the subject. In certain embodiments, the second
dosage is equal to the first dosage when the PAA:PAGN ratio is 1 to
2.5, i.e., falling within the target range. In certain embodiments,
the target range is divided into one or more subranges. In certain
of these embodiments, the second dosage may be equal to the first
dosage if the PAA:PAGN ratio is 1 to 1.5 or 2 to 2.5, but the
subject may be subjected to more frequent monitoring. In certain
other embodiments, the second dosage may be greater than the first
dosage if the PAA:PAGN ratio is 1 to 1.5 or 1 to 2 and the subject
has recently exhibited particularly acute symptoms of a nitrogen
retention disorder or another condition for which PAA prodrug
administration is expected to be beneficial. Similarly, the second
dosage may be less than the first dosage if the PAA:PAGN ratio is
greater than 1.5 or 2 but not greater than 2.5, depending on the
subject's specific characteristics. In certain embodiments, the
increase or decrease in the second dosage versus the first dosage
depends on the precise plasma PAA:PAGN ratio. For example, where
the plasma PAA:PAGN ratio is only slightly less than 1, the dosage
may be increased only slightly, but where the PAA:PAGN ratio is
significantly less than 1, the dosage may be increased more.
Similarly, the decrease in dosage for subjects exhibiting a ratio
above 2.5 may vary depending on how far above 2.5 the ratio
extends. In certain embodiments, measurement of plasma PAA and PAGN
ratio takes place after the PAA prodrug has had sufficient time to
reach steady state (e.g., 48 hours, 48 to 72 hours, 72 hours to 1
week, 1 week to 2 weeks, or greater than 2 weeks after PAA prodrug
administration). In certain embodiments, the above steps may be
repeated until a desired plasma PAA:PAGN ratio (e.g., 1 to 2.5 or 1
to 2) is achieved. For example, the methods may comprise measuring
plasma PAA and PAGN levels after administration of the second
dosage, calculating the plasma PAA:PAGN ratio, determining whether
the PAA prodrug dosage needs to be adjusted based on whether the
PAA:PAGN ratio falls within the target range, and administering a
third dosage of the PAA prodrug.
[0042] In certain embodiments, methods are provided for treating a
nitrogen retention disorder or a condition for which PAA prodrug
administration is expected to be beneficial in a subject that has
not previously been administered a PAA prodrug. These methods
comprise administering a first dosage of a PAA prodrug, measuring
plasma PAA and PAGN levels, calculating the plasma PAA:PAGN ratio,
determining whether the PAA prodrug dosage needs to be adjusted
based on whether the PAA:PAGN ratio falls within a target range,
and administering a second dosage of the PAA prodrug. In certain
embodiments, the target range for PAA:PAGN ratio is 1 to 2.5 or 1
to 2. In certain of these embodiments, the second dosage is greater
than the first dosage if the PAA:PAGN ratio is less than 1 (i.e.,
the dosage is increased) and less than the first dosage if the
PAA:PAGN ratio is greater than 2.5 (i.e., the dosage is decreased).
In other embodiments, the second dosage may or may not be greater
than the first dosage if the PAA:PAGN ratio is less than 1,
depending on one or more additional characteristics of the subject.
In certain embodiments, the second dosage is equal to the first
dosage when the PAA:PAGN ratio is 1 to 2.5, i.e., falling within
the target range. In certain embodiments, the target range is
divided into one or more subranges. In certain of these
embodiments, the second dosage may be equal to the first dosage if
the PAA:PAGN ratio is 1 to 1.5 or 2 to 2.5, but the subject may be
subjected to more frequent monitoring. In certain other
embodiments, the second dosage may be greater than the first dosage
if the PAA:PAGN ratio is 1 to 1.5 or 1 to 2 and the subject has
recently exhibited particularly acute symptoms of a nitrogen
retention disorder or another condition for which PAA prodrug
administration is expected to be beneficial. Similarly, the second
dosage may be less than the first dosage if the PAA:PAGN ratio is
greater than 1.5 or 2 but not greater than 2.5, depending on the
subject's specific clinical or biochemical characteristics. In
certain embodiments, the increase or decrease in the second dosage
versus the first dosage depends on the precise plasma PAA:PAGN
ratio. For example, where the plasma PAA:PAGN ratio is only
slightly less than 1, the dosage may be increased only slightly,
but where the PAA:PAGN ratio is significantly less than 1, the
dosage may be increased more. Similarly, the decrease in dosage for
subjects exhibiting a ratio above 2.5 may vary depending on how far
above 2.5 the ratio extends. In certain embodiments, measurement of
plasma PAA and PAGN ratio takes place after the PAA prodrug has had
sufficient time to reach steady state (e.g., 48 hours, 48 to 72
hours, 72 hours to 1 week, 1 week to 2 weeks, or greater than 2
weeks after PAA prodrug administration). In certain embodiments,
the above steps may be repeated until a desired plasma PAA:PAGN
ratio (e.g., 1 to 2.5 or 1 to 2) is achieved. For example, the
methods may comprise measuring plasma PAA and PAGN levels after
administration of the second dosage, calculating the plasma
PAA:PAGN ratio, determining whether the PAA prodrug dosage needs to
be adjusted based on whether the PAA:PAGN ratio falls within the
target range, and administering a third dosage of the PAA
prodrug.
[0043] A method of administering a PAA prodrug to a subject with a
nitrogen retention disorder or another condition for which PAA
prodrug administration is expected to be beneficial. These methods
comprise administering a first dosage of the PAA prodrug, measuring
plasma PAA and PAGN levels, calculating the plasma PAA:PAGN ratio,
determining whether the PAA prodrug dosage needs to be adjusted
based on whether the PAA:PAGN ratio falls within a target range,
and administering a second dosage of the PAA prodrug. In certain
embodiments, the target range for PAA:PAGN ratio is 1 to 2.5 or 1
to 2. In certain of these embodiments, the second dosage is greater
than the first dosage if the PAA:PAGN ratio is less than 1 (i.e.,
the dosage is increased) and less than the first dosage if the
PAA:PAGN ratio is greater than 2.5 (i.e., the dosage is decreased).
In other embodiments, the second dosage may or may not be greater
than the first dosage if the PAA:PAGN ratio is less than 1,
depending on one or more additional characteristics of the subject.
In certain embodiments, the second dosage is equal to the first
dosage when the PAA:PAGN ratio is 1 to 2.5, i.e., falling within
the target range. In certain embodiments, the target range is
divided into one or more subranges. In certain of these
embodiments, the second dosage may be equal to the first dosage if
the PAA:PAGN ratio is 1 to 1.5 or 2 to 2.5, but the subject may be
subjected to more frequent monitoring. In certain other
embodiments, the second dosage may be greater than the first dosage
if the PAA:PAGN ratio is 1 to 1.5 or 1 to 2 and the subject has
recently exhibited particularly acute symptoms of a nitrogen
retention disorder or another condition for which PAA prodrug
administration is expected to be beneficial. Similarly, the second
dosage may be less than the first dosage if the PAA:PAGN ratio is
greater than 1.5 or 2 but not greater than 2.5, depending on the
subject's specific biochemical or clinical characteristics. In
certain embodiments, the increase or decrease in the second dosage
versus the first dosage depends on the precise plasma PAA:PAGN
ratio. For example, where the plasma PAA:PAGN ratio is only
slightly less than 1, the dosage may be increased only slightly,
but where the PAA:PAGN ratio is significantly less than 1, the
dosage may be increased more. Similarly, the decrease in dosage for
subjects exhibiting a ratio above 2.5 may vary depending on how far
above 2.5 the ratio extends. In certain embodiments, measurement of
plasma PAA and PAGN ratio takes place after the PAA prodrug has had
sufficient time to reach steady state (e.g., 48 hours, 48 to 72
hours, 72 hours to 1 week, 1 week to 2 weeks, or greater than 2
weeks after PAA prodrug administration). In certain embodiments,
the above steps may be repeated until a desired plasma PAA:PAGN
ratio (e.g., 1 to 2.5 or 1 to 2) is achieved. For example, the
methods may comprise measuring plasma PAA and PAGN levels after
administration of the second dosage, calculating the plasma
PAA:PAGN ratio, determining whether the PAA prodrug dosage needs to
be adjusted based on whether the PAA:PAGN ratio falls within the
target range, and administering a third dosage of the PAA
prodrug.
[0044] In certain embodiments, methods are provided herein for
achieving a target plasma PAA:PAGN ratio in a subject with a
nitrogen retention disorder or another condition for which PAA
prodrug administration is expected to be beneficial. These methods
comprise administering a first dosage of a PAA prodrug, measuring
plasma PAA and PAGN levels, calculating the plasma PAA:PAGN ratio,
determining whether the PAA prodrug dosage needs to be adjusted
based on whether the PAA:PAGN ratio falls within a target range,
and administering a second dosage of the PAA prodrug based on the
PAA:PAGN ratio. If the PAA:PAGN ratio is above the target range,
the second dosage is less than the first dosage. If the PAA:PAGN
ratio is below the target range, the second dosage is greater than
the first dosage. These steps are repeated until a target plasma
PAA:PAGN ratio is achieved. In certain embodiments, the target
ratio falls within a target range of 1 to 2.5 or 1 to 2. In certain
embodiments, the increase or decrease in the second dosage versus
the first dosage depends on the precise plasma PAA:PAGN ratio. For
example, where the plasma PAA:PAGN ratio is only slightly less than
1, the dosage may be increased only slightly, but where the
PAA:PAGN ratio is significantly less than 1, the dosage may be
increased more. Similarly, the decrease in dosage for subjects
exhibiting a ratio above 2.5 may vary depending on how far above
2.5 the ratio extends. In certain embodiments, measurement of
plasma PAA and PAGN ratio takes place after the PAA prodrug has had
sufficient time to reach steady state (e.g., 48 hours, 48 to 72
hours, 72 hours to 1 week, 1 week to 2 weeks, or greater than 2
weeks after PAA prodrug administration).
[0045] In certain embodiments, methods are provided for evaluating
the dosage of a PAA prodrug in a subject who has previously been
administered a first dosage of a PAA prodrug. These methods
comprise measuring plasma PAA and PAGN levels, calculating the
plasma PAA:PAGN ratio, and determining whether the first dosage of
the PAA prodrug is effective based on whether the PAA:PAGN ratio
falls within a target range. In certain embodiments, the target
range for PAA:PAGN ratio is 1 to 2.5 or 1 to 2. In certain of these
embodiments, the first dosage is considered too low if the PAA:PAGN
ratio is less than 1, and too high if the PAA:PAGN ratio is greater
than 2.5. In other embodiments, the first dosage is considered
potentially too low if PAA:PAGN ratio is less than 1, with a final
decision depending on one or more additional characteristics of the
subject. In certain embodiments, the target range is divided into
one or more subranges. In certain of these embodiments, the first
dosage is considered potentially effective if the PAA:PAGN ratio is
1 to 1.5 or 2 to 2.5, but the subject may be subjected to more
frequent monitoring. In certain other embodiments, the first dosage
may be considered too low if the PAA:PAGN ratio is 1 to 1.5 or 1 to
2 and the subject has recently exhibited particularly acute
symptoms of a nitrogen retention disorder or another condition for
which PAA prodrug administration is expected to be beneficial.
Similarly, in certain embodiments the first dosage may be
considered too high if the PAA:PAGN ratio is greater than 1.5 or 2
but not greater than 2.5, depending on the subject's specific
biochemical or clinical characteristics. In certain embodiments,
measurement of plasma PAA and PAGN ratio takes place after the PAA
prodrug has had sufficient time to reach steady state (e.g., 48
hours, 48 to 72 hours, 72 hours to 1 week, 1 week to 2 weeks, or
greater than 2 weeks after PAA prodrug administration). In certain
embodiments, the methods further comprise a step of administering a
second dosage that differs from the first dosage, and in certain of
these embodiments the above steps may be repeated until a desired
plasma PAA:PAGN ratio (e.g., 1 to 2.5 or 1 to 2) is achieved. For
example, the methods may comprise administering a second dosage
that differs from the first dosage, measuring plasma PAA and PAGN
levels after administration of the second dosage, calculating the
plasma PAA:PAGN ratio, and determining whether the second dosage of
the PAA prodrug is effective based on whether the PAA:PAGN ratio
falls within a target range.
[0046] In certain embodiments, methods are provided for adjusting
the dosage of a PAA prodrug in a subject who has previously been
administered a first dosage of a PAA prodrug. These methods
comprise measuring plasma PAA and PAGN levels, calculating the
plasma PAA:PAGN ratio, and determining whether to adjust the dosage
of the PAA prodrug based on whether the PAA:PAGN ratio falls within
a target range. In certain embodiments, the target range for
PAA:PAGN ratio is 1 to 2.5 or 1 to 2. In certain of these
embodiments where the target range is 1 to 2.5, a PAA:PAGN ratio of
less than 1 indicates the PAA prodrug dosage needs to be adjusted
upwards, while a PAA:PAGN ratio above 2.5 indicates the PAA prodrug
dosage needs to be adjusted downwards. In other embodiments, a
PAA:PAGN ratio of less than 1 indicates that the PAA prodrug dosage
potentially needs to be adjusted upwards, with a final decision
depending on one or more additional characteristics of the subject.
In certain embodiments, the target range is divided into one or
more subranges. In certain of these embodiments, a PAA:PAGN ratio
of 1 to 1.5 or 2 to 2.5 indicates that the dosage need not be
adjusted, but that the subject should be subjected to more frequent
monitoring. In certain other embodiments, a PAA:PAGN ratio of 1 to
1.5 or 1 to 2 indicates that the dosage needs to be increased when
the subject has recently exhibited particularly acute symptoms of a
nitrogen retention disorder or another condition for which PAA
prodrug administration is expected to be beneficial. Similarly, in
certain embodiments a PAA:PAGN ratio greater than 1.5 or 2 but not
greater than 2.5 may indicate that the dosage needs to be
decreased, depending on the subject's specific biochemical or
clinical characteristics. In certain embodiments, measurement of
plasma PAA and PAGN ratio takes place after the PAA prodrug has had
sufficient time to reach steady state (e.g., 48 hours, 48 to 72
hours, 72 hours to 1 week, 1 week to 2 weeks, or greater than 2
weeks after PAA prodrug administration). In certain embodiments
where a determination is made that the dosage needs to be adjusted,
the methods further comprise a step of administering a second
dosage that differs from the first dosage, and in certain of these
embodiments the above steps may be repeated until a desired plasma
PAA:PAGN ratio (e.g., 1 to 2.5 or 1 to 2) is achieved. For example,
the methods may comprise administering a second dosage that differs
from the first dosage, measuring plasma PAA and PAGN levels after
administration of the second dosage, calculating the plasma
PAA:PAGN ratio, and determining whether the second dosage of the
PAA prodrug needs to be adjusted based on whether the PAA:PAGN
ratio falls within a target range. In certain embodiments, the
increase or decrease in the second dosage versus the first dosage
depends on the precise plasma PAA:PAGN ratio. For example, where
the plasma PAA:PAGN ratio is only slightly less than 1, the dosage
may be increased only slightly, but where the PAA:PAGN ratio is
significantly less than 1, the dosage may be increased more.
Similarly, the decrease in dosage for subjects exhibiting a ratio
above 2.5 may vary depending on how far above 2.5 the ratio
extends.
[0047] In certain embodiments, methods are provided for optimizing
the therapeutic efficacy of a PAA prodrug for use in treating a
nitrogen retention disorder in a subject. These methods comprise
measuring plasma PAA and PAGN levels in a subject who has
previously been administered a PAA prodrug, calculating the plasma
PAA:PAGN ratio, determining whether to adjust the dosage of the PAA
prodrug based on whether the PAA:PAGN ratio falls within a target
range, and administering an adjusted dosage of the PAA prodrug as
necessary. These steps are repeated until the subject exhibits a
plasma PAA:PAGN ratio falling within the target range (e.g., 1 to
2.5 or 1 to 2). In certain embodiments where the target range is 1
to 2.5, a plasma PAA:PAGN ratio of less than 1 indicates that the
dosage needs to be adjusted upwards, while a ratio greater than 2.5
indicates that the dosage needs to be decreased. In certain
embodiments, the target range is divided into one or more
subranges. In certain of these embodiments, a PAA:PAGN ratio of 1
to 1.5 or 2 to 2.5 indicates that the dosage does not need to be
adjusted, but that the subject should be subjected to more frequent
monitoring. In certain other embodiments, a PAA:PAGN ratio of 1 to
1.5 or 1 to 2 indicates that the dosage needs to be increased when
the subject has recently exhibited particularly acute symptoms of a
nitrogen retention disorder or another condition for which PAA
prodrug administration is expected to be beneficial. Similarly, in
certain embodiments a PAA:PAGN ratio greater than 1.5 or 2 but not
greater than 2.5 may indicate that the dosage needs to be
decreased, depending on the subject's specific biochemical or
clinical characteristics. In certain embodiments, measurement of
plasma PAA and PAGN ratio takes place after the PAA prodrug has had
sufficient time to reach steady state (e.g., 48 hours, 48 to 72
hours, 72 hours to 1 week, 1 week to 2 weeks, or greater than 2
weeks after PAA prodrug administration). In certain embodiments,
the magnitude of the increase or decrease in dosage may be based on
the precise PAA:PAGN ratio. For example, a PAA:PAGN ratio that is
slightly less than 1 may indicate that the dosage needs to be
increased slightly, while a ratio significantly less than 1 may
indicate the dosage needs to be increased to a greater degree. In
certain embodiments, the above steps are repeated until the subject
exhibits a PAA:PAGN ratio falling within the target range.
[0048] In certain embodiments, methods are provided for determining
whether a prescribed first dosage of a PAA prodrug can be safely
administered to a subject. These methods comprise administering the
prescribed first dosage to the subject, measuring plasma PAA and
PAGN levels, calculating the plasma PAA:PAGN ratio, and determining
whether the prescribed first dosage is safe for the subject based
on whether the PAA:PAGN ratio falls above a target range, wherein a
PAA:PAGN ratio falling above the target range indicates that the
first dosage cannot be or potentially cannot be safely administered
to the subject. In certain embodiments, the target range for
PAA:PAGN ratio is 1 to 2.5 or 1 to 2. In certain of these
embodiments where the target range is 1 to 2.5, a PAA:PAGN ratio
above 2.5 indicates the PAA prodrug dosage is unsafe and needs to
be adjusted downwards. In certain embodiments, the target range is
divided into one or more subranges. In certain of these
embodiments, a PAA:PAGN ratio of 2 to 2.5 indicates that the first
dosage is safe, but that the subject should be subjected to more
frequent monitoring. In other embodiments, a PAA:PAGN ratio of 2 to
2.5 indicates that the first dosage is potentially unsafe, with a
final determination of safety taking into account the subject's
specific biochemical or clinical characteristics. In certain
embodiments, measurement of plasma PAA and PAGN ratio takes place
after the PAA prodrug has had sufficient time to reach steady state
(e.g., 48 hours, 48 to 72 hours, 72 hours to 1 week, 1 week to 2
weeks, or greater than 2 weeks after PAA prodrug administration).
In certain embodiments where a determination is made that the first
dosage is unsafe and needs to be decreased, the methods further
comprise a step of administering a second dosage that is lower than
the first dosage, and in certain of these embodiments the above
steps may be repeated until a desired plasma PAA:PAGN ratio (e.g.,
1 to 2.5 or 1 to 2) is achieved. For example, the methods may
comprise administering a second dosage that is lower than the first
dosage, measuring plasma PAA and PAGN levels after administration
of the second dosage, calculating the plasma PAA:PAGN ratio, and
determining whether the second dosage of the PAA prodrug can be
safely administered to the subject based on whether the PAA:PAGN
ratio falls above a target range.
[0049] In certain embodiments, methods are provided for determining
whether a prescribed first dosage of a PAA prodrug will be
effective for treating a nitrogen retention disorder or another
disorder for which PAA prodrug administration is expected to be
beneficial. These methods comprise administering the prescribed
first dosage to the subject, measuring plasma PAA and PAGN levels,
calculating the plasma PAA:PAGN ratio, and determining whether the
prescribed first dosage will be effective for the subject based on
whether the PAA:PAGN ratio falls below a target range, wherein a
PAA:PAGN ratio falling below the target range indicates that the
first dosage will not be or potentially will not be effective for
treating a disorder. In certain embodiments, the target range for
PAA:PAGN ratio is 1 to 2.5 or 1 to 2. In certain of these
embodiments where the target range is 1 to 2.5, a PAA:PAGN ratio
below 1 indicates the PAA prodrug dosage is unlikely to be
effective and needs to be adjusted upwards. In other embodiments, a
PAA:PAGN ratio below 1 indicates that the first dosage is
potentially ineffective, with a final determination of whether the
dosage is likely to be ineffective based on the subject's specific
biochemical or clinical characteristics. In certain embodiments,
the target range is divided into one or more subranges. In certain
of these embodiments, a PAA:PAGN ratio of 1 to 1.5 indicates that
the first dosage is likely to be effective, but that the subject
should be subjected to more frequent monitoring. In other
embodiments, a PAA:PAGN ratio of 1 to 1.5 indicates that the first
dosage is potentially ineffective, with a final determination of
whether the dosage is likely to be ineffective taking into account
the subject's specific biochemical or clinical characteristics. In
certain embodiments, measurement of plasma PAA and PAGN ratio takes
place after the PAA prodrug has had sufficient time to reach steady
state (e.g., 48 hours, 48 to 72 hours, 72 hours to 1 week, 1 week
to 2 weeks, or greater than 2 weeks after PAA prodrug
administration). In certain embodiments where a determination is
made that the first dosage is likely to be ineffective and needs to
be increased, the methods further comprise a step of administering
a second dosage that is higher than the first dosage, and in
certain of these embodiments the above steps may be repeated until
a desired plasma PAA:PAGN ratio (e.g., 1 to 2.5 or 1 to 2) is
achieved. For example, the methods may comprise administering a
second dosage that is higher than the first dosage, measuring
plasma PAA and PAGN levels after administration of the second
dosage, calculating the plasma PAA:PAGN ratio, and determining
whether the second dosage of the PAA prodrug is likely to be
ineffective for treating a disorder based on whether the PAA:PAGN
ratio falls above a target range.
[0050] Provided herein in certain embodiments are methods for
monitoring therapy with a PAA prodrug in patients with a nitrogen
retention disorder. These methods comprise administering a PAA
prodrug to the subject, measuring plasma PAA and PAGN levels, and
calculating the plasma PAA:PAGN ratio. In these methods, a PAA:PAGN
ratio falling within a target range (e.g., 1 to 2.5 or 1 to 2)
indicates that the therapy is effective, while a ratio falling
outside this range indicates that the therapy may need to be
adjusted. In certain embodiments, the plasma PAA:PAGN ratio is
compared to a previously obtained PAA:PAGN ratio from the same
subject to evaluate the effectiveness of PAA prodrug
administration.
[0051] In certain embodiments, the methods provided herein may be
used in conjunction with the methods described in WO09/134460 and
WO10/025303. In these embodiments, urinary PAGN levels may be
determined in addition to plasma PAA:PAGN ratio, with both
measurements being used to evaluate or adjust PAA prodrug
dosage.
[0052] A "PAA prodrug" as used herein refers to any drug that
contains or is converted to PAA following administration to a
subject, or to any pharmaceutically acceptable salt, ester, acid,
or derivative thereof. A PAA prodrug may be administered via any
route, including oral or parenteral administration. A PAA prodrug
may be converted directly to PAA (e.g., a salt or ester of PAA; PBA
or a salt or ester thereof such as NaPBA), or it may be converted
to PAA via an intermediate (e.g., a pre-prodrug such as HPN-100).
Other examples of PAA prodrugs include
butyroyloxymethyl-4-phenylbutyrate.
[0053] An adjustment to the dosage of a PAA prodrug as discussed
herein may refer to a change in the amount of drug per
administration (e.g., an increase from a first dosage of 3 mL to a
second dosage of 6 mL), a change in the number of administration
within a particular time period (e.g., an increase from once a day
to twice a day), or any combination thereof.
[0054] A "subject in need thereof" as used herein refers to any
individual having a condition or suspected of having a condition
for which administration of a PAA prodrug is expected to be
beneficial. For example, a subject may be an individual with a
nitrogen retention disorder or suspected of having a nitrogen
retention disorder, including for example UCD, HE, and/or kidney
failure/ESRD (Lee 2010; McGuire 2010; Lichter 2011). Likewise, a
subject may have or be suspected of having another condition for
which PAA prodrug administration is expected to be beneficial,
including for example cancer (Thiebault 1994; Thiebault 1995),
neurodegenerative disorders such as Huntington's Disease (Hogarth
2007), amyotrophic lateral sclerosis (ALS) (Cudkowicz 2009), and
spinal muscular atrophy (SMA) (Mercuri 2004; Brahe 2005), metabolic
disorders (e.g., maple syrup urine disease (MSUD) (Bruneti-Pieri
2011), or sickle cell disease (Hines 2008).
[0055] A subject that has previously been administered a PAA
prodrug may have been administered the drug for any duration of
time sufficient to reach steady state. For example, the subject may
have been administered the drug over a period of 2 to 7 days, 1
week to 2 weeks, 2 weeks to 4 weeks, 4 weeks to 8 weeks, 8 weeks to
16 weeks, or longer than 16 weeks.
[0056] A "PAA prodrug" as used herein refers to any drug that
contains or is converted to PAA following administration to a
subject, or to any pharmaceutically acceptable salt, ester, acid,
or derivative thereof. A PAA prodrug may be administered via any
route, including oral or parenteral administration. A PAA prodrug
may be converted directly to PAA (e.g., PBA or a salt thereof such
as NaPBA), or it may be converted to PAA via an intermediate (e.g.,
a pre-prodrug such as HPN-100). Other examples of PAA prodrugs
include butyroyloxymethyl-4-phenylbutyrate.
[0057] An adjustment to the dosage of a PAA prodrug as discussed
herein may refer to a change in the amount of drug per
administration (e.g., an increase from a first dosage of 3 mL to a
second dosage of 6 mL), a change in the number of administration
within a particular time period (e.g., an increase from once a day
to twice a day), or any combination thereof.
[0058] The terms "treat," "treating," or "treatment" as used herein
may refer to preventing a disorder, slowing the onset or rate of
development of a disorder, reducing the risk of developing a
disorder, preventing or delaying the development of symptoms
associated with a disorder, reducing or ending symptoms associated
with a disorder, generating a complete or partial regression of a
disorder, or some combination thereof. For example, where the
disorder being treated is a nitrogen retention disorder, "treating"
may refer to lowering waste nitrogen levels below a threshold
level, preventing waste nitrogen levels from reaching a threshold
level, decreasing the likelihood of waste nitrogen levels exceeding
a threshold level, reducing or ending symptoms associated with
elevated waste nitrogen levels, or a combination thereof.
[0059] With regard to the methods of treatment disclosed herein,
interpretation of the PAA:PAGN ratio must be performed in the
context of the therapeutic objective. For example, in subjects
being treated for a nitrogen retention disorder, the therapeutic
objective is elimination of waste nitrogen in the form of PAGN. In
subjects being treated for other disorders for which PAA prodrug
administration is expected to be beneficial (e.g.,
neurodegenerative disorders, MSUD), the therapeutic objective is
safely achieving target plasma levels of PAA and/or PBA.
[0060] Any methods known in the art may be used to obtain a plasma
blood sample. For example, blood from a subject may be drawn into a
tube containing heparin or ethylenediaminetetraacetic acid (EDTA).
In certain embodiments, the sample can be placed on ice and
centrifuged to obtain plasma within 15 minutes of collection,
stored at 2-8.degree. C. (36-46.degree. F.) and analyzed within 3
hours of collection. In other embodiments, the blood plasma sample
is snap frozen, stored at .ltoreq.-18.degree. C. (.ltoreq.0.degree.
F.) and analyzed at a later time. For example, the sample may be
analyzed at 0-12 hours, 12-24 hours, 24-48, 48-96 hours after
freezing, or within any other timeframe over which the sample has
demonstrated stability. In certain of these embodiments, the blood
sample is stored at a temperature between 0-15.degree. C., such as
2-8.degree. C. In other embodiments, the blood sample is stored
below 0.degree. C. or below -18.degree. C.
[0061] Measurement of PAA and PAGN levels in a plasma sample is
carried out using techniques known in the art. For example, PAA and
PAGN levels may be measured using liquid chromatography/mass spec
analyses.
[0062] Any combination of embodiments described herein can be
envisioned. Although individual features may be included in
different claims, these may be advantageously combined.
[0063] The following examples are provided to better illustrate the
claimed invention and are not to be interpreted as limiting the
scope of the invention. To the extent that specific materials are
mentioned, it is merely for purposes of illustration and is not
intended to limit the invention. One skilled in the art may develop
equivalent means or reactants without the exercise of inventive
capacity and without departing from the scope of the invention. It
will be understood that many variations can be made in the
procedures herein described while still remaining within the bounds
of the present invention. It is the intention of the inventors that
such variations are included within the scope of the invention.
EXAMPLES
Example 1
Analysis of PAA:PAGN Ratio in UCD and HE Subjects
[0064] Plasma PAA and PAGN levels and PAA:PAGN ratio were analyzed
in more than 4000 plasma samples obtained from various clinical
trials of healthy adults, severely hepatic impaired adults with
clinically decompensated Child-Pugh B or C cirrhosis, and UCD
patients ages 29 days or older. Healthy and hepatically impaired
adults received HPN-100, while UCD subjects received both HPN-100
and NaPBA. Clinical trial populations are summarized in Tables 1
and 2.
TABLE-US-00001 TABLE 1 Clinical studies and analysis populations
Study Protocols Analysis Group Description Demographics Included
Populations 1 Short-term (<=2-4 weeks) Adults and children UP
1204-003 A, B exposure in UCD subjects ages 29 days or
HPN-100-005SO greater (N = 81) HPN-100-006 HPN-100-012 2 Long-term
exposure in Adults and children HPN-100-005SE A UCD and HE subjects
ages 6 years or HPN-100-007 greater (N = 180) HPN-100-008 Part B 3
Short-term (<=4 weeks) Adults (N = 15) HPN-100-008 Part A A, B
exposure in hepatic impaired subjects 4 Short-term exposure (<=4
Adults (N = 98) HPN-100-010 A, B weeks) in healthy subjects
TABLE-US-00002 TABLE 2 Demographics and number of samples used No.
of No. of time-specific No. of sample points PK sample points
subjects (Population A) (Population B) Attribute Count Percent
Count Percent Count Percent Population Healthy 86 17.0 2126 34.4
2126 38.5 Hepatic 103 20.4 830 13.4 830 15.0 Encephalopathy (HE)
UCD 158 31.3 1616 26.1 1281 23.2 Total 347 100.0 4572 100.0 4237
100.0 Age 29 days-<6 yrs 15 4.3 110 2.4 110 2.6 6-<18 yrs 47
13.5 373 8.2 213 5.0 18+ yrs 285 82.1 4089 89.4 3914 92.4 Sex F 199
57.3 2394 52.4 2152 50.8 M 148 42.7 2178 47.6 2085 49.2
[0065] Analysis Population A consisted of quantifiable levels of
PAA and PAGN metabolites derived from all studies described above.
All PAA and PAGN levels used for analysis came from blood samples
drawn once dosing with NaPBA or HPN-100 had reached steady state.
Analysis Population B consisted of quantifiable levels of PAA and
PAGN metabolites during studies in which pharmacokinetics were
analyzed and for which blood draws were performed over 12 or 24
hours at steady state and for which the timing of the blood sample
in relation to dosing was known. Subjects in study groups 1, 3 and
4 above contributed to these points. Analysis Population B was the
source of analyses that examined how PAA levels changed with time
relative to dosing, where dosing could have been with either NaPBA
or HPN-100. To be eligible for Analysis Population B, the time of
the blood draw relative to the time of initiation of dosing during
the dosing period had to have been recorded.
[0066] Data on metabolite levels were pooled across a wide range of
age levels--infants, toddlers, children, adolescents, and adults.
All children, defined as ages under 18, were UCD patients. The
majority of the blood sampling points came from adults (89.4%).
Newborn infants (<29 days old) were not studied in any of the
clinical trials for the investigational agent HPN-100. The
population of blood sampling points were roughly equally divided
between female and male (57.3% female, 42.7% male).
[0067] To examine the predictive ability of PAA:PAGN ratios, a
subject was considered to have achieved a high value of PAA if any
PAA value up to 24 hours since initiation of dosing equaled or
exceeded 400 .mu.g/mL or equaled or exceeded 500 .mu.g/mL. PAA:PAGN
ratios were grouped into one of three categorization schemes: a.)
[0-<=2.0], [>2.0], b.) [0-<=2.5, >2.5], c.)
[0-<=3.0, >3.0]. The repeated measures categorical outcome
was modeled using GEE with a logit link function, ratio category as
the independent variable, and SUBJECTID as the repeated measures
factor. Confidence intervals for the predicted probabilities were
computed by bootstrap estimation of 1000 resamplings of the
original data, as detailed in Davison & Hinkley, "Bootstrap
Methods and Their Application," Cambridge Univ. Press (1997), pp.
358-362.
[0068] Results are summarized in FIGS. 2-5. A striking curvilinear
relationship was observed between plasma PAA levels and PAA:PAGN
ratio at any given timepoint. FIG. 2A shows the relationship
between the ratio of PAA:PAGN concentrations and absolute PAA
levels in micrograms per milliliter among blood samples that had
quantifiable values for both PAA and PAGN. The ratio axis (i.e. `X`
axis) is plotted on a logarithmic (base e) scale. For ratios less
than 1.0, increases in ratio are not associated with
correspondingly elevated or increased levels of PAA. Above ratios
of 1.0, there is a gradual increase in PAA levels, and a noticeable
upswing in PAA levels that begins in the vicinity of a ratio of
2.0. This finding suggests that when the ratio of PAA precursor to
PAGN product approaches higher values, the values of PAA are also
correspondingly high. This increase in the ratio of precursor (PAA)
to product (PAGN) implies ineffective PAA to PAGN conversion,
regardless of whether the PAA is derived from HPN-100 or NaPBA.
[0069] To determine whether excessive PAA build-up is a function of
dosing, the plots mentioned above were repeated, but this time
adjusting for assigned dose level of NaPBA or HPN-100 at the time
of the blood draw. Since the UCD population consisted of a mixture
of children and adults undergoing both short-term therapy and
long-term therapy, total assigned daily dose for UCD patients was
standardized to body surface area and reported in PBA-equivalent
grams meter.sup.2. Healthy and HE subjects were all adults and
their assigned dose was not adjusted by body surface area. Dose
levels for healthy and HE subjects were reported in HPN-100
equivalent mL. Dose levels for UCD subjects were reported in
NaPBA-equivalent grams.
[0070] The excess of PAA over PAGN, indicated by larger ratios as
PAA increases, was evident across all dosage groups, disease
populations, and types of treatment in UCD patients (i.e., applies
to both NaPBA and HPN-100). This finding suggests that analysis of
the precursor (PAA) to product (PAGN) ratio may be predictive of
the efficiency of conversion among patients with or without liver
dysfunction (UCD patients have normal liver function apart from
their urea cycle dysfunction) and independently of dose. As a
corollary, the presence of liver dysfunction (e.g. cirrhosis) by
itself, is not necessarily a reliable determinant of whether a
particular patient is at risk for high PAA levels.
[0071] The ability of PAA:PAGN ratios to predict extremely high
plasma PAA concentrations was determined by modeling the
probability that a subject would exceed a PAA value of 400 or 500
.mu.g/mL anytime during a 24 hour dosing period, based on the ratio
of PAA to PAGN computed at pre-dose (presumably trough), 12 hours
after dosing (presumably peak), and the maximum ratio encountered
anytime between pre-dose and 12 hours post-dose. This interval of
0-12 hours was chosen for practical reasons, as it would encompass
the entire interval corresponding to the usual outpatient
visit.
[0072] Since subjects could have multiple dosing periods within a
given clinical study, the probability was modeled using Generalized
Estimating Equations. Three categorizations of ratios were modeled:
a.) [0-<=2.0] [>2.0], b.) [0-<=2.5, >2.5], c.)
[0-<=3.0, >3.0]. The models were repeated with PAA values
greater than or equal to 500 .mu.g/mL considered extreme. Results
are summarized in Table 3.
TABLE-US-00003 TABLE 3 Probabilities of extreme PAA values
encountered during 24 hour PK sampling with PAA:PAGN ratios (all
subjects combined) Probability that Time of a Subject With
Bootstrapped Blood Draw Observed This Ratio Will 95% PAA Value
Considered Used For Ratio Ratio of Exceed High Confidence High
Classification PAA/PAGN Value* (%) Interval** [<=2.0, >2.0]
>=400 .mu.g/mL t = 0 (fasting) <=2.0 0.005 (0.5%) 0.004,
0.020 >2.0 0.164 (16.4%) 0.041, 0.281 t = 12 hours <=2.0
0.003 (0.3%) 0.004, 0.021 >2.0 0.227 (22.7%) 0.048, 0.412
MAX(0-12) <=2.0 0.002 (0.2%) 0.004, 0.010 >2.0 0.143 (14.3%)
0.036, 0.263 >=500 .mu.g/mL t = 0 (fasting) <=2.0 did not
converge >2.0 t = 12 hours <=2.0 did not converge >2.0
MAX(0-12) <=2.0 did not converge >2.0 [<=2.5, >2.5]
>=400 .mu.g/mL t = 0 (fasting) <=2.5 0.008 (0.8%) 0.004,
0.023 >2.5 0.191 (19.1%) 0.053, 0.366 t = 12 hours <=2.5
0.007 (0.7%) 0.004, 0.016 >2.5 0.364 (36.4%) 0.125, 0.752
MAX(0-12) <=2.5 0.003 (0.3%) 0.004, 0.013 >2.5 0.200 (20.0%)
0.050, 0.381 >=500 .mu.g/mL t = 0 (fasting) <=2.5 0.003
(0.3%) 0.004, 0.011 <=2.5 0.084 (8.4%) 0.029, 0.214 t = 12 hours
>2.5 did not converge >2.5 MAX(0-12) <=2.5 did not
converge >2.5 [<=3, >3] >=400 .mu.g/mL t = 0 (fasting)
<=3.0 0.010 (1.0%) 0.004, 0.025 >3.0 0.205 (20.5%) 0.059,
0.398 t = 12 hours <=3.0 0.013 (1.3%) 0.004, 0.028 >3.0 0.250
(25.0%) 0.113, 0.576 MAX(0-12) <=3.0 0.003 (0.3%) 0.004, 0.014
>3.0 0.229 (22.9%) 0.059, 0.438 >=500 .mu.g/mL t = 0
(fasting) <=3.0 0.003 (0.3%) 0.004, 0.010 >3.0 0.102 (10.2%)
0.032, 0.255 t = 12 hours <=3.0 did not converge >3.0
MAX(0-12) <=3.0 did not converge >3.0 Analysis repeated for
each ratio cut off category independently. *Probability derived
from Generalized Estimating Equations model with logit link
function. **Confidence interval derived from method disclosed in
Davison & Hinkley, "Bootstrap Methods and Their Application,"
Cambridge Univ. Press (1997), pp. 358-362, using 1000 re-samplings
of original data.
[0073] Because of the sparseness of samples in which PAA equaled or
exceeded 500 .mu.g/mL, 400 .mu.g/mL proved to be a more stable and
predictable target (i.e. high) value. Of the three categorizations
of ratio considered, the cutpoint of 2.5 was the best discriminator
and predictor of the risk of experiencing an high value. For
example, referring to Table 3, a subject with a PAA:PAGN ratio
>2.5 at t=12 hours after dosing has a 36.4% chance (95% c.
i.=0.125, 0.752) of exceeding 400 .mu.g/mL in PAA sometime during
the 24-hour PK sampling period.
[0074] Results were similar whether the ratio was computed from
plasma drawn at pre-dose, 12 hours after initiation of dosing, or
the maximum ratio encountered anytime between pre-dose and 12 hours
after initiation of dosing.
[0075] Due to the very high intra-day variability of plasma PAA
levels, a PAA:PAGN ratio observed as exceeding 2.0 at a certain
time following dosing may not remain greater than 2.0 in subsequent
times. To evaluate the optimal time for obtaining a PAA:PAGN ratio
measurement (i.e., the time that gives the greatest probability of
correctly detecting a subject whose PAA:PAGN ratio ever equals or
exceeds 2.0 during the dosing period), ratios were evaluated at 0
(pre-dose) and 2, 4, 6, 8, 10, and 12 hours post-dosing and modeled
using GEE methodology. Pairwise differences in sensitivity between
time points were evaluated using LS means and confidence intervals
were computed.
[0076] FIG. 3 plots the estimated probabilities of correctly
detecting a ratio profile that ever equals of exceeds 2.0. With the
exception of time=2 hours and time=10 hours, time points of 0, 4,
6, 8, and 12 hours post-dosing were equally effective in detecting
subjects who equal or exceed a PAA:PAGN ratio of 2.0 at some point
during the dosing period. Sensitivities were in the range of 75-90
percent. There were too few blood samples collected at t=10 hours
to analyze inter-time differences. Differences in predictive value
were observed. For example, blood samples collected at t=2 hours
post-dosing had a significantly lower probability of detecting
subjects who equal or exceed a PAA:PAGN ratio of 2.0 than samples
collected at t=0 (p=0.036), 4 (p=0.032), or 6 hours (p=0.017)
post-dosing (p values are comparisons of t=2 hour probability with
other time points). Similarly, a sample collected at t=12 hours
following initiation of dosing had the highest probability (87%) of
detecting a subject whose ratio ever equals or exceeds 2.0.
However, for practical clinical purposes, the differences in
predictive value among time points was trivial relative to the
dramatically greater variability in PAA values themselves, meaning
that random blood draws can be used for measurement of PAA:PAGN
ratio.
[0077] Further exploration of the fluctuation of PAA:PAGN ratios
over time was conducted by dividing the subject population into
cohorts according to the maximum PAA:PAGN ratio achieved during the
24-hour PK sampling time during the dosing period. Cohorts were
divided into "low" (maximum ratio <=2.0), "medium" (maximum
ratio: 2.01-2.50), and "high" (maximum ratio >2.50). Each cohort
was then followed over time during the dosing period at t=0
hours(pre-dose), 4, 6, and 8 hours post-dosing and the distribution
of PAA:PAGN ratios within the cohort summarized using a
box-and-whisker plot at each time point. This analysis was
conducted for the PK-timepoint-specific population as a whole
(analysis population B) as well as for each disease subpopulation
separately.
[0078] FIG. 4 plots the progression of ratios for all subjects
combined. Each "panel" of the plot that divides the graphing space
into thirds represents one cohort. Subjects in the high cohort had
high ratios throughout the day and not only at a particular time
point. Therefore, subjects in this cohort (n=73 subject/dosing
periods) started with high ratios (median ratio >2.5) and
remained high throughout the first 12 hours. This finding is
consistent with the findings plotted in FIG. 3 which revealed the
consistency of sensitivity in ratios.
[0079] The relationship between PAA levels and PAA:PAGN ratios was
further analyzed by categorizing ratios into "low" (maximum ratio
<=2.0), "medium" (maximum ratio: 2.01-2.50), and "high" (maximum
ratio >2.50). Unlike the previous analysis, this analysis did
not associate subject/dosing periods with particular cohorts (i.e.,
all samples and all time points are combined with regard to the
subject or dosing period).
[0080] FIG. 5A shows the box-and-whisker plots of PAA levels
grouped by the above categories of PAA:PAGN ratio for all subjects,
while FIG. 5B shows the same for UCD and HE subjects only. The
results were very similar in both analysis sets. Following a
statistically significant overall Kruskal-Wallis test
(p<0.0001), pairwise comparisons of PAA levels were conducted
using Wilcoxon-Mann-Whitney with a Bonferroni alpha correction of
(0.0167). In both analysis sets, ratios greater than 2.5 had
significantly higher PAA levels (p<0.001) than either ratios
between 2.0-2.5 or ratios less than 2.0. Furthermore, ratios
between 2.0-2.5 were associated with significantly higher PAA
levels than ratios less than 2.0 (p<0.001).
Example 2
Analysis of PAA:PAGN Ratio as a Guide to Dose Adjustment and
Monitoring in a UCD Patient
[0081] Patient 1 was a 15 year old partial OTC female receiving
HPN-100 as maintenance therapy for her UCD at a dose of 9 mL/day.
The patient's ammonia had been controlled since her last routine
visit around 6 months ago, but she was complaining of headache and
lack of appetite for the past 3 days. Ammonia and metabolite levels
were tested after overnight fasting and showed the following
results: ammonia 55 .mu.mol/L, PAA and PAGN below levels of
quantification. The physician suspected non-compliance with drug
and repeated the tests in midday several hours after lunch and
found the following results: ammonia: 117 .mu.mol/L; PAA 55
.mu.g/L, PAGN 121 .mu.g/L, and PAA:PAGN ratio approximately 0.5.
The patient indicated that she had been fully compliant with her
medication. Based on the PAA to PAGN ratio of 0.5 and ammonia of
117, the physician decided to increase the dosage of HPN-100 to 12
mL/day. After one week of treatment with the new dose of HPN-100,
all symptoms resolved and the laboratory tests after overnight
fasting showed the following: ammonia 9 .mu.mol/L; PAA 12.9
.mu.g/L, PAGN of 9 .mu.g/L, and PAA:PAGN ratio of 1.3. Midday tests
showed the following: ammonia 35 .mu.mol/L, PAA 165 .mu.g/L, PAGN
130 .mu.g/L, and PAA:PAGN ratio of .about.1.2. The patient was
considered controlled and the dose remained at 12 mL/day.
Example 3
Analysis of PAA:PAGN Ratio as a Guide to Dose Adjustment in a UCD
Patient
[0082] Patient 2 was a 1 year old male OTC receiving 600 mg/kg of
NaPBA per day. The patient presented with poor feeding and
somnolence. Laboratory tests showed ammonia levels of <9
.mu.mol/L, PAA levels of 530 .mu.g/L, PAGN levels of 178 .mu.g/L,
and a PAA:PAGN ratio of >2.5, suggesting that the dose of NaPBA
was greater than the patient could effectively convert to PAGN. The
treating physician decided to decrease the dose of NaPBA to 450
mg/Kg/day. After one week of treatment with the new dosage, the
patient's mother reported that he was eating well and was no longer
somnolent. Laboratory tests showed the following: ammonia 20
.mu.mol/L, PAA 280 .mu.g/L, and PAGN 150 .mu.g/L.
Example 4
Analysis of PAA:PAGN Ratio as a Guide to Assessment of Importance
of a High PAA Level in a UCD Patient
[0083] Patient 3 is a 25 year old OTC female who is being treated
with HPN-100. The physician had to increase the dose of HPN-100
several times in order to achieve clinical and blood ammonia within
normal limits. Patient 3 was treated at a dose of 18 mL/day for her
UCD for the past month. In her next office visit, she did not have
any complaints and the following lab results were reported: ammonia
22 .mu.mol/L, PAA 409 .mu.g/L, PAGN 259 .mu.g/L, and PAA:PAGN ratio
of 1.5. Despite the patient's relatively high PAA levels, the
PAA:PAGN ratio indicated that the subject was being adequately
treated and that the patient was able to effectively metabolize the
high dose of HPN-100 that she was receiving. The physician decided
to continue the treatment as planned.
Example 5
Analysis of PAA:PAGN Ratio as a Guide to Dose Adjustment in a
Patient With Spinal Muscular Atrophy and Concomitant Liver
Disease
[0084] Patient 4 was a 2 year old female being treated with a
liquid form of NaPBA for her type II SMA. The patient also suffered
from chronic hepatitis C virus infection acquired perinatally from
her infected mother. The patient had been having mild to moderate
elevation of transaminases since birth, with episodes of icterus
and a recent liver biopsy has confirmed presence of chronic
hepatitis and cirrhosis. The patient was receiving 4 g of NaPBA per
day, and the physician wanted to increase the dosage due to the
patient's growth but was concerned about the effects of liver
dysfunction on drug metabolism. The physician ordered plasma PAA
and PAGN levels and the results were as follows: PAA 110 .mu.g/L,
PAGN 85 .mu.g/L, PAA:PAGN ratio of 1.2. The physician decided to
increase the dosage of NaPBA to 6 g/day, and repeated the plasma
metabolite level measurements after one week of treatment with the
new regimen. The results were as follows: PAA 155 .mu.g/L, PAGN 110
.mu.g/L, and PAA:PAGN ratio of 1.4. The physician decided to leave
the patient on 6 g/day of NaPBA since his liver seems to have
adequate capacity to metabolize 6 g of NaPBA.
Example 6
Analysis of PAA:PAGN Ratio as a Guide to Dose Adjustment in a
Patient With Huntington's Disease and Concomitant Liver Disease
[0085] Patient 5 was a 56 year old male diagnosed with Huntington's
disease several years ago. He also had a history of alcohol abuse
and was diagnosed with alcoholic cirrhosis last year. His wife
enrolled him in clinical trials that involved an experimental drug
delivering PBA at a slow rate, thereby enabling once-a-day dosing
of the drug. The study had an option for dose escalation after 2
weeks of treatment if clinically safe. Although the protocol did
not exclude patients with liver dysfunction, the investigator was
concerned about PBA metabolism and possible accumulation of PAA in
higher doses due to the patient's liver dysfunction. The
investigator enrolled the patient in the low dose group and
performed plasma PBA, PAA and PAGN measurements after 6 weeks of
treatment with experimental drug. The patient reported improvement
in his HD symptoms with no specific complains. Plasma metabolite
levels after six weeks of treatment were as follows: PBA 45
.mu.g/L; PAA 159 .mu.g/L, and PAGN 134 .mu.g/L. The dosage of the
drug was increased by 50%. After four days of treatment at the new
dosage, the patient started to complain about short episodes of
somnolence. The investigator performed a blood test and observed
the following: PBA 44 .mu.g/L; PAA 550 .mu.g/L, PAGN 180 .mu.g/L,
and PAA:PAGN ratio of >3. The PAA:PAGN ratio of greater than 2.5
indicated that the patient's liver could not effectively metabolize
the higher dose of the drug, and the investigator therefore decided
to reduce the dosage of the experimental drug and not continue dose
escalation.
Example 7
Analysis of PAA:PAGN Ratio as a Guide to Dose Adjustment in a
Patient With MSUD
[0086] Patient 6 was a 4 year old female being treated with HPN-100
for MSUD. The patient was receiving 6 mL of HPN-100 once a day, and
the physician wanted to increase the dosage due to the patient's
growth. Midday plasma PAA and PAGN measurements after the dose of
medication were as follows: PAA 550 .mu.g/L, PAGN 180 .mu.g/L, and
PAA:PAGN ratio of >2.5. The physician believed a lower dosage of
HPN-100 would not be as effective for the patient, and decided to
change the dosing regimen to 3 mL BID instead of 6 mL QD based on
the high PAA:PAGN ratio. The tests were repeated after one week of
treatment with the new BID regimen, with the following results: PAA
350 .mu.g/L, PAGN 190 .mu.g/L, and PAA:PAGN ratio of 1.8. Based on
the ratio of 1.8, the physician decided to leave the patient on 3
mL BID since she can efficiently use a total dose of 6 mL/day given
in divided doses but not as a bolus.
Example 8
Analysis of PAA:PAGN Ratio as a Guide to Monitor a Patient With HE
and Hepatic Impairment
[0087] Patient 7 was a 55 year old Caucasian male diagnosed with
alcoholic cirrhosis 3 years ago. His transaminase levels had been
mildly elevated and he had recently experienced mild episodes of
HE. In the last assessment at the time of hospital admission for a
grade 2 HE episode, the patient had a blood ammonia of 85
.mu.mol/L, ALT of 55 U/L, and AST of 47 U/L, and a calculated MELD
score of 11. The physician decided to start an ammonia scavenging
therapy for the patient and treated him with HPN-100 6 mL BID. The
patient returned for a follow up visit after 3 months, during which
time he had experienced no episodes of HE. His laboratory
assessments showed the following: ammonia of 30 .mu.mol/L, plasma
PAA level of 285 .mu.g/mL, PAGN level of 120 .mu.g/L, ALT of 66
U/L, AST of 50 U/L, and calculated MELD score of 13. The physician
suspected that the patient's hepatic function may be deteriorating
and was concerned about possible accumulation of PAA. She
calculated the ratio of PAA to PAGN as 2.4, and confirmed that the
patient had not experienced any unusual symptoms such as dizziness,
headache, or nausea. Considering patient's ammonia control, lack of
specific side effects, and clinical remission, the physician
decided not to change the dose and to see the patient in two weeks
to repeat the laboratory tests. The physician also warned the
patient to call her immediately if he experienced any of these
symptoms. In two weeks, the patient's laboratory assessments were
essentially unchanged from the previous visit, with a PAA to PAGN
ratio of 2.3, and the patient did not report any unusual symptoms.
Based on the PAA:PAGN ratio of less than 2.5, the physician decided
to continue dosing with 6 mL BID of HPN-100 until the next routine
visit.
Example 9
Analysis of PAA:PAGN Ratio as a Guide to Monitoring Treatment in a
Patient With Parkinson's Disease
[0088] HPN-100 treatment was initiated at a dose of 4 mL twice a
day in a patient with Parkinson's Disease to produce target
circulating levels of PAA expected to produce clinical benefit.
After one week of treatment, the patient's circulating PAA level of
50 .mu.g/mL was below the target range, and the PAA:PAGN ratio was
determined to be 0.9. The physician concluded that the HPN-100 dose
could be safely adjusted upward, and the dose was increased by 50%
to 6 mL BID. The PAA level and PAA/PAGN ratio one week later were
found to be 75 .mu.g/mL and 1.4, respectively. Since 75 .mu.g/mL
was still below the therapeutic PAA target level and the PAA:PAGN
ratio of 1.4 indicated that conversion of PAA to PAGN had not been
saturated, the patient's dosage was increased again by 50% to 9 mL
BID. One week later, the patient's PAA and PAA:PAGN ratio were
found to be 159 .mu.g/mL and 2.6, respectively. Since the target
PAA level was now approximately therapeutic but the PAA:PAGN ratio
indicated that PAA to PAGN conversion was approaching saturation,
HPN-100 dosage was decreased to 8 mL BID, at which time the
patient's circulating PAA level was determined to be close to the
target range and his PAA:PAGN ratio was determined to be 2. The
patient's dose was not further adjusted and he continued to be
monitored.
[0089] As stated above, the foregoing is merely intended to
illustrate various embodiments of the present invention. The
specific modifications discussed above are not to be construed as
limitations on the scope of the invention. It will be apparent to
one skilled in the art that various equivalents, changes, and
modifications may be made without departing from the scope of the
invention, and it is understood that such equivalent embodiments
are to be included herein. All references cited herein are
incorporated by reference as if fully set forth herein.
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