Note: Descriptions are shown in the official language in which they were submitted.
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METHODS AND FORMULATIONS FOR TREATING SIALIC ACID
DEFICIENCIES
CROSS-REFERENCE TO RELATED APPLICATIONS
100011 This application claims the benefit of priority of U.S. Provisional
Application No.
61/588,069, filed on January 18, 2012 and entitled "Methods and Formulations
for Treating
Sialic Acid Deficiencies", and .U.S. Provisional Application No. 61/709,549,
filed on October
4, 2012 and entitled "Methods and Formulations for Treating Sialic Acid
Deficiencies". The
content of these applications are herein incorporated by reference in their
entireties for all
purposes.
BACKGROUND
100021 Sialic acid (SA) is a sugar with a net negative charge. It is often
present on
terminating branches of N-glycans, 0-glycans, and glycosphingolipids
(gangliosides), and
occasionally capping side chains of GPI anchors. Sialic acid modification of
cell surface
molecules plays a role in many biological phenomena such as protein structure
stability,
regulation of cell adhesion, and signal transduction.. Sialic acid deficiency
disorders such as
Hereditary Inclusion Body Myopathy (HIBM or HIBM type 2), Nonaka myopathy, and
Distal Myopathy with Rimmed Vacuoles (DMRV) are clinical diseases resulting
from. a
reduction in sialic acid production.
[00031 HIBM is a rare autosomal recessive neuromuscular disorder caused by a
biosynthetic
defect in the sialic acid synthesis pathway. Eisenberg et al., Nat. Genet.
29:83-87 (2001). The
disease usually manifests between the ages of 20 to 40 such as foot drop and
slowly
progressive muscle weakness and atrophy. Patients may suffer difficulties
walking with foot
drop, gripping, use of hands, and swallowing. The disease is progressive; most
afflicted
individuals become incapacitated and wheelchair-confined within two to three
decades. No
treatments are available.
[00041 Studies of an Iranian-Jewish genetic isolate mapped the mutation
associated with
HIBM to chromosome 9p12-13. Argov et al., Neurology 60:1519-1523 (2003). The
causative
mutations were identified for HIBM in the gene GIVE, which encodes the
bifunctional
enzyme LTDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase
(GNE/MNK).
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Eisenberg et al., Nat. Genet. 29:83-87 (2001). DMRV is a Japanese variant,
allelic to HIBM.
Nishino etal., Neurology 59:1689-1693 (2002).
100051 The biosynthesis steps and feedback regulation of GNE/MNK is depicted
in Figure 1.
The production of sialic acid on glycoconjugates requires the conversion of N-
acetylglucosamine (conjugated to its carrier nucleotide sugar IMP) to sialic
acid. The sialic
acid subsequently enters the nucleus where it is conjugated with its
nucleotide sugar carrier
CMP to make CMP-sialic acid, which is used as a donor sugar for glycosylation
reactions in
the cell. CMP-sialic acid is a knovvn regulator of GNE/MNK activity. Jay
etal., Gene Reg. &
Sys. .Biol. 3:181-190 (2009). Patients with HIBM have a deficiency in the
production of sialic
acid by the GNE/MNK enzyme, which is involved in the first two steps of this
sequence.
Nearly twenty ONE mutations have been reported in HIBM patients from different
ethnic
backgrounds with founder effects among the Iranian Jews and Japanese.
Broccolini et al.,
Hum. Mutat. 23:632 (2004).
[00061 Because the production of sialic acid is the key reason the mutation
causes the
disease, replacing a metabolite after the genetic block in the pathway could,
in theory,
alleviate symptoms of a sialic acid deficiency. Jay et al., Gene Reg. and Sys.
Biology 3:181-
190 (2009). In practice, however, administering one or more compounds in the
sialic acid
biosynthetic pathway in vivo is a significant challenge. These compounds have
extraordinarily rapid clearance rates and are excreted in the urine before
they can be
metabolized.
SUMMARY OF THE INVENTION
[00071 The present invention provides method for treating a sialic acid
deficiency in an
individual in need thereof comprising orally administering a sialic acid, or a
pharmaceutically
acceptable salt, solvate, or ester thereof, wherein the method provides a
therapeutically
effective amount of sialic acid over a period of greater than about four
hours.
[00081 In some embodiments of the present invention, the sialic acid, or a
pharmaceutically
acceptable salt, solvate, or ester thereof, is in an extended release
formulation. In some
embodiments of the present invention, the sialic acid, or a pharmaceutically
acceptable salt,
solvate, or ester thereof, is in both an extended release formulation and an
immediate release
formulation.
[00091 In some embodiments of the present invention, the method provides a
mean Cmin
sialic acid of at least about 0.11 mcgirn1 at steady state during the dosing
intervals.
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10010) In some embodiments of the present invention, the method provides a
mean plasma
concentration of sialic acid of at least about 0.16 mcg/m.I at steady state
during the dosing
intervals.
[00111 In some embodiments the present invention, the method provides a mean
plasma
concentration of sialic acid at steady state during the dosing intervals that
is at least about
50% higher than the mean plasma concentration of sialic acid in the individual
before the
administration of the sialic acid, or a pharmaceutically acceptable salt,
sol.vate, or ester
thereof.
[00121 In some embodiments of the present invention, the method provides an.
improved
absorption profile when the extended release formulation is administered under
fed
conditions than being administered under fasting conditions.
[00131 In some embodiments of the methods, the sialic acid deficiency is a
myopathy
associated with sialic acid deficiency.
100141 In some embodiments of the methods, the sialic acid deficiency is a
myopathy
associated with sialic acid deficiency. In some embodiments, the myopathy
associated with
sialic acid deficiency is Hereditary Inclusion Body Myopathy (H1 BM), Nonaka
myopathy,
and/or Distal Myopathy with Rimmed Vacuoles (DMRV).
[00151 In some embodiments of the methods, the extended release formulation is
in a solid
matrix form.
BRIEF DESCRIPTION OF FIGURES
100161 Figure 1 provides a diagram of intracellular sialic acid metabolism.
[00171 Figure 2 shows the particle size distribution for sialic acid.
[00181 Figure 3 shows the particle size distribution plot for ProCR sialic
acid 250 mg final
blends.
100191 Figure 4 shows the dissolution plot of sialic acid 250 and 325 mg
sustained release
(SR) tablets by direct compression.
[00201 Figure 5 shows the dissolution profile of sialic acid 325 and 500 mg
sustained release
(SR) uncoated tablets.
[00211 Figure 6 shows the dissolution profile of sialic acid 325 and 500 mg
sustained release
(SR) coated tablets.
[00221 Figure 7 shows the dissolution profile of ManNAc 325 mg tablets.
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100231 Figure 8 shows the individual concentrations of sialic acid versus time
in beagle dog
serum following IV or oral administration. (A and B) concentration after
administration of
TA-1 capsules; (C) concentration after administration of TA-2 tablets; (D)
concentration after
administration of TA-3 tablet; (E) concentration after administration of TA-4
tablets; (F)
concentration after administration of TA-5 tablets; (G and
concentration after intravenous
administration of TA-6.
[00241 Figure 9 shows pharmacokinetic data for single doses of SA-ER versus SA
API in the
canine - sialic acid in serum crossover study of orally administered SA-ER
tablets.
[00251 Figure 10 shows results from repeated dosing of SA-ER. in the dog: Day
0 to Day 7.
Group average (Gp average) sialic acid concentration in serum orally
administered SA-ER
tablets (1625 mg TID). Some degree of accumulation occurred over the seven day
period.
[00261 Figure 11 shows a comparison of urinary SA excretion in a crossover
study of orally
administered SA-ER in the dog - individual canine excretion levels over a 24
hour period.
Three orally administered dose levels versus API versus the last day of 7 days
of dosing.
[00271 Figure 12 shows a mean total urinary SA excretion comparison - total
sialic acid in
urine during 24 hours after/during dosing. Single doses, compared with API and
with
seventh day of repeat dosing.
[00281 Figure 13 shows the Study Scheme associated with the ER.-SA. human
clinical trial of
Example 7.
[00291 Figure 14 shows pharmacokinetic data obtained for single dose ER-SA
administration
(650 mg) for six different human patients. Open circles represent the first
day in which
baseline monitoring was conducted; closed circles represent the next day when
the fasted
single dose was administered. Free sialic acid concentration is in pg/mL
serum.
[00301 Figure 15 shows pharmacokinetic data obtained for single dose ER-SA
administration
(1,950 mg) for six different human patients. Open circles represent the first
day in which
baseline monitoring was conducted; closed circles represent the next day when
the fasted
single dose was administered. Free sialic acid concentration is in pg/rnL
serum.
[00311 Figure 16 shows pharmacokinetic data obtained for single dose ER-SA
administration
(2,925 mg) for two different human patients. Open circles represent the first
day in which
baseline monitoring was conducted; closed circles represent the next day when
the fasted
single dose was administered. Free sialic acid concentration is in figimiL
serum.
[00321 Figure 17 shows pharmacokinetic data obtained for single dose ER.-SA.
administration
(4,825 mg) for one human patient. Open circles represent the first day in
which baseline
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monitoring was conducted; closed circles represent the next day when the
fasted single dose
was administered. Free sialic acid concentration is in pg/mI., serum..
[00331 Figure 18 shows pharm.acokinetic data obtained for ER-S.A repeated
administration
(650 x 3; 1,950 mg) for six different human patients. Open circles represent
the first day in
which baseline monitoring was conducted; closed circles represent data from
the last day of 7
days of three times per day divided dosing. Free sialic acid concentration is
in 1.1g/niL serum.
[00341 Figure 19 shows pharmacokinetic data obtained for ER-SA repeated
administration
(975 x 3; 2,925 mg) for five different human patients. Open circles represent
the first day in
which baseline monitoring was conducted; closed circles represent data from
the last day of 7
days of three times per day divided dosing. Free sialic acid concentration is
in pg/inL serum.
[00351 Figure 20 is a schematic of the Phase I Interim Safety Study dosing
schedule.
[00361 Figure 21 shows graphs depicting the mean free sialic acid
concentrations at single
dose levels (fasted and fed states). The individual mean PK curves are shown
for the 650 mg,
1950 mg, 2925 mg, 4875 mg and 6000 mg dose levels to compare fasted, fed and
baseline SA
levels. Panels for 650 mg, 1950 mg, 2925 mg and 6000 mg have the same size y-
axis but the
4875 mg panel has a larger y-axis due to the higher levels achieved. Baseline
curves for the
subjects in each cohort are shown as open circles, fasted levels as black
circles and fed curves
as grey circles. Baseline sialic acid levels for day 1 for a group of subjects
(both Fasted and
Fed states) are presented graphically on the same axis so that easy visual
comparison can be
made of before treatment to after treatment sialic acid levels. The 6000 mg
group was
administered the 500 mg tablet whereas the rest of the dose groups were
administered the 325
mg tablet.
[00371 Figure 22 is a graph depicting mean free sialic acid concentrations at
different single
dose levels (fasted state). The mean PK curves and standard deviations are
shown for the 650
mg, 1950 mg, 2925 mg, 4875 mg and 6000 mg dose levels (Fasted state). The 6000
mg
group was administered the 500 mg tablet whereas the rest of the dose groups
were
administered the 325 mg tablet.
[0038i Figure 23 is a graph depicting mean free sialic acid concentrations at
different single
dose levels (fed state). The mean PK curves and standard deviations are shown
for the 650
mg, 1950 mg, 2925 mg, 4875 mg and 6000 mg dose levels (Fed state). The 6000 mg
group
was administered the 500 mg tablet whereas the rest of the dose groups were
administered the
325 mg tablet.
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100391 Figures 24A-F show graphs depicting free sialic acid concentrations in
serum at
different repeat dose levels.
100401 Figure 25 shows a graph depicting free sialic acid concentrations in
serum by 4 days
of dosing sialic acid extended release formulation followed by 4 days of
dosing sialic acid
extended release formulation plus sialic acid immediate release formulation.
[00411 Figure 26 shows a graph depicting free sialic acid concentrations in
serum by 4 days
of dosing sialic acid extended release formulation followed by 4 days of
dosing sialic acid
extended release formulation plus sialic acid immediate release formulation.
DETAILED DESCRIPTION
[00421 The present application provides extended release pharmaceutical
formulations
comprising one or more compounds in the sialic acid biosynthetic pathway or
derivative
thereof and methods of treating and preventing sialic acid deficiencies
utilizing the extended
release pharmaceutical formulations. This invention concerns designing an
approach to
substrate replacement that provides individuals with sialic acid deficiencies
stable and steady
day and nighttime replacement without high concentration spikes across a broad
range of
genotypes and in multiple tissues. This invention can optimally achieve this
substrate
replacement and treatment benefit through the combination of using extended
release
formulations and one or more metabolites, including combinations of
m.etabolites.
[00431 It is understood that the description refers to and includes effective
amounts of an
active agent, such as the compounds provided herein, which include but are not
limited to the
compounds included under the heading "Therapeutic Agent." Thus, it is
understood that any
of the extended release formulations detailed herein may comprise an effective
amount of a
therapeutic agent, such as an effective amount of sialic acid, or a
pharmaceutically acceptable
salt thereof.
Definitions
[00441 The terms "oral administration" and "oral ingestion" refer to all
conventional forms
for the oral delivery of a pharmaceutical composition to an individual and
that result in the
deposition of the pharmaceutical formulation into the gastrointestinal tract
(including the
gastro portion of the gastrointestinal tract, i.e., the stomach) of the
patient. Accordingly, oral
administration and oral ingestion include, by way of example, actual ingestion
of a solid or
liquid pharmaceutical composition, oral gavage, and the like.
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100451 The terms "treating" and "treatment" as used herein refer to an
approach for obtaining
beneficial or desired results including clinical results. For purposes of this
invention,
beneficial or desired clinical results include, but are not limited to, one or
more of the
following: decreasing the severity and/or frequency one or more symptoms
resulting from the
disease, diminishing the extent of the disease, stabilizing the disease (e.g.,
preventing or
delaying the worsening of the disease), delay or slowing the progression of
the disease,
ameliorating the disease state, increasing production of sialic acid, the
sial.ylation precursor
CMP-sialic acid (e.g., increasing intracellular production of sialic acid) and
restoring the
level of sialylation in muscle and other proteins, decreasing the dose of one
or more other
medications required to treat the disease, and/or increasing the quality of
life. "Treating" a
patient with a formulation described herein includes management of an
individual to inhibit
or cause regression of a disease or condition.
[00461 "Prophylaxis" or "prophylactic treatment" "or preventive treatment"
refers to
prevention of the occurrence of symptoms and/or their underlying cause, for
example,
prevention of a disease or condition in a patient susceptible to developing a
disease or
condition (e.g., at a higher risk, as a result of genetic predisposition,
environmental factors,
predisposing diseases or disorders, or the like). Prophylaxis includes HIBM
myopathy in
which chronic disease changes in the muscles are irreversible and for which
animal model
data suggests treatment benefit in prophylaxis.
[00471 As used herein, "delaying" the progression of the disease means to
defer, hinder,
slow, retard, stabilize, and/or postpone development of the disease. This
delay can be of
varying lengths of time, depending on the history of the disease and/or
individual being
treated.
[00481 As used herein, an "at risk" individual is an individual who is at risk
of developing a
sialic acid deficiency. An individual "at risk" may or may not have detectable
disease, and
may or may not have displayed detectable disease prior to the treatment
methods described
herein. "At risk" denotes that an individual has one or more so-called risk
factors, which are
measurable parameters that correlate with development of a sialic acid
deficiency, which are
described herein. An individual having one or more of these risk factors has a
higher
probability of developing a sialic acid deficiency than an individual without
these risk
factor(s).
100491 The term "effective amount" refers to the amount of one or more
compounds in the
sialic acid biosynthetic pathway in a sufficient amount to render a desired
treatment outcome.
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An effective amount may be comprised within one or more doses, i.e., a single
dose or
multiple doses may be required to achieve the desired treatment endpoint.
100501 A "therapeutically effective amount" refers to an amount of one or more
compounds
in the sialic acid biosynthetic pathway sufficient to produce a desired
therapeutic outcome
(e.g., reduction of severity of a disease or condition). In one embodiment,
the therapeutically
effective amount refers to a therapeutically effective plasma concentration of
sialic acid. A
"prophylactically effective amount" refers to an amount of a pharmaceutical
formulation
including one or more compounds in the sialic acid biosynthetic pathway
sufficient to prevent
or reduce severity of a future disease or condition when administered to an
individual who is
susceptible and/or who may develop a disease or condition.
[00511 The term "extended release" refers to a drug-containing formulation or
fraction
thereof, in which release of the drug is not immediate, i.e., with an
"extended release"
formulation, administration does not result in immediate release of the drug
into an
absorption pool. In general, the term "extended release" as used herein
includes controlled
release, sustained release, and delayed release formulations.
[00521 By "pharmaceutically acceptable" is meant a material that is not
biologically or
otherwise undesirable, i.e., the material may be incorporated into a
pharmaceutical
composition administered to a patient without causing any significant
undesirable biological
effects or interacting in a deleterious manner with any of the other
components of the
composition in which it is contained. When the term. "pharmaceutically
acceptable" is used to
refer to a pharmaceutical carrier or excipient, it is implied that the carrier
or excipient has met
the required standards of toxicological and manufacturing testing or that it
is included on the
Inactive Ingredient Guide prepared by the U.S. Food and Drug administration.
[00531 The term "disorder" or "disease" used interchangeably herein, refers to
any alteration
in the state of the body or one of its organs and/or tissues, interrupting or
disturbing the
performance of organ function and/or tissue function (e.g., causes organ
dysfunction) and/or
causing a symptom such as discomfort, dysfunction, distress, or even death to
a subject
afflicted with the disease.
[00541 The term "individual" or "patient" refers to an animal, for example, a
mammal and
includes, but is not limited to, human, bovine, horse, feline, canine, rodent,
or primate.
Preferably, the individual is a human.
100551 The term "derivative" as used herein includes derivatives, analogs,
prodrugs, and
unnatural precursors.
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[0056] The term "pharmaceutically acceptable salt" refers to a salt which
retains the
biological effectiveness of the compound and which is not biologically or
otherwise
undesirable.
[0057] Reference to "about" a value or parameter herein includes (and
describes) variations
that are directed to that value or parameter per se. For example, description
referring to
"about X" includes description of "X".
100581 As used herein and in the appended claims, the singular forms "a,"
"or," and "the"
include plural referents unless the context clearly dictates otherwise. It is
understood that
embodiments, aspects and variations of the invention described herein include
"comprising,"
"consisting" and/or "consisting essentially of' embodiments, aspects and
variations.
[0059] Pharmacokinetic parameters describe the in vivo characteristics of the
active agent,
i.e., the free sialic acid over time, such as plasma concentration (C), C.,
C., C24, T., and
AUC. "C." is the measured concentration of the active agent in the plasma at
the point of
maximum concentration. "C." is the measured concentration of an active agent
in the plasma
at about n hours after administration. "C24" is the measured concentration of
an active agent
in the plasma at about 24 hours after administration. The term 'fn..," refers
to the time at
which the measured concentration of an active agent in the plasma is the
highest after
administration of the active agent. "AUC" is the area under the curve of a
graph of the
measured concentration of an active agent (typically plasma concentration) vs.
time,
measured from one time point to another time point. For example AUCo.., is the
area under
the curve of plasma concentration versus time from time 0 to time t. The
AUC0,, or AUCO-
INF is the calculated area under the curve of plasma concentration versus time
from. time 0 to
time infinity.
Extended Release Formulations
[0060] Provided herein are extended release pharmaceutical formulations
comprising as the
therapeutic agent one or more compounds in the sialic acid biosynthetic
pathway or a
derivative thereof or a pharmaceutically acceptable salt of the foregoing. In
one embodiment,
the extended release pharmaceutical formulations comprise a therapeutic agent
as detailed
herein and a polymer. An extended release formulation comprising a therapeutic
agent and a
polymer may further comprise one or more additional components, such as any
one or more
of a diluent, an excipient, an antioxidant, a lubricant, a colorant, a binder,
a disintegrant, and
the like. It is understood that reference to and description of extended
release pharmaceutical
formulations comprising one or more compounds in the sialic acid biosynthetic
pathway or a
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derivative thereof below is exemplary and that this description applies
equally to and includes
extended release pharmaceutical formulations comprising any one or more
compounds in the
sialic acid biosynthetic pathway. It is also understood that reference to and
description of
extended release pharmaceutical formulations comprising any one or more
derivatives of
compounds in the sialic acid biosynthetic pathway below is exemplary and that
this
description applies equally to and includes extended release pharmaceutical
formulations
comprising any one or more derivatives, analogs, prodrugs, and/or unnatural
precursor
compounds in the sialic acid biosynthetic pathway.
[00611 In one variation, the extended release formulation comprises sialic
acid, or a
pharmaceutically acceptable salt thereof, as the therapeutic agent. In another
variation, the
extended release formulation comprises MaNAc, or a pharmaceutically acceptable
salt
thereof, as the therapeutic agent. In another variation, the extended release
formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof and
MaNAc, or a
pharmaceutically acceptable salt thereof, as the therapeutic agents. In
another variation, the
extended release formulation comprises a prodrug of one or more compounds in
the sialic
acid biosynthetic pathway such as a prodrug of sialic acid, or a
pharmaceutically acceptable
salt thereof, as the therapeutic agent. In another variation, the extended
release formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof, as the
therapeutic agent
and further comprises a hydrocolloid polymer, an anionic, pH-dependent gel
forming co-
polymer and a water swellable, pH independent polymer and optionally further
comprises a
lubricant and/or an excipient. In a particular variation, the extended release
formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof,
carrageenan, sodium
alginate and either hypromellose or polyethylene oxide. In a further
variation, the extended
release formulation comprises sialic acid, or a pharmaceutically acceptable
salt thereof,
carrageenan, sodium alginate, either hypromellose or polyethylene oxide,
magnesium stearate
and rnicrocrystalline cellulose and colloidal silicon dioxide. In one aspect,
the extended
release formulation is a formulation of Table E. In yet another aspect, the
extended release
formulation is a formulation of Table 8. In yet another aspect, the extended
release
formulation is a formulation of Example 6. In yet another aspect, the extended
release
formulation is a formulation of Example 7.
Therapeutic Agent
100621 It is believed that administration of sialic acid or a compound in the
sialic acid
biosynthetic pathway, or a derivative thereof, or a pharmaceutically
acceptable salt of any of
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the foregoing, may be administered as a therapeutic agent (e.g., as substrate
replacement) to
an individual who has or is suspected of having a sialic acid deficiency
disorder. Extended
release formulations comprising such compounds, or pharmaceutically acceptable
salts
thereof, as the therapeutic agent are provided herein. In one aspect, the
sialic acid or a
compound in the sialic acid biosynthetic pathway, or a derivative thereof, or
a
pharmaceutically acceptable salt of any of the foregoing, is sialic acid or a
pharmaceutically
acceptable salt thereof. In one aspect, any of the extended release
formulations detailed
herein may comprise an effective amount of a therapeutic agent, such as an
effective amount
of sialic acid or a pharmaceutically acceptable salt thereof.
100631 A compound in the sialic acid biosynthetic pathway or a derivative
thereof in one
variation is a compound, or pharmaceutically acceptable salt thereof, that is
at or downstream
from ManNAc in the sialic acid biosynthetic pathway. In a particular
variation, the
therapeutic agent is a compound, or pharmaceutically acceptable salt thereof,
that is at or
downstream from ManNAc in the sialic acid biosynthetic pathway and is depicted
in Figure
1.
[00641 A compound in the sialic acid biosynthetic pathway or a derivative
thereof in another
variation is a compound, or a pharmaceutically acceptable salt thereof, that
is at or upstream
from CMP-sialic acid in the sialic acid biosynthetic pathway. In a particular
variation, the
therapeutic agent is a compound, or a pharmaceutically acceptable salt
thereof, that is at or
upstream. from CM.P-sialic acid in the sialic acid biosynthetic pathway and is
depicted in
Figure 1. In one such variation, the compound in the sialic acid biosynthetic
pathway or a
derivative thereof does not include glucose or a pharmaceutically acceptable
salt thereof.
100651 In a particular variation, the compound in the sialic acid biosynthetic
pathway or a
derivative thereof in one variation is a compound, or a pharmaceutically
acceptable salt
thereof, that is: (i) at or downstream from ManNAc in the sialic acid
biosynthetic pathway,
and (ii) is at or upstream from CMP-sialic acid in the sialic acid
biosynthetic pathway. In one
such variation, the compound is a compound depicted in Figure 1, or a
pharmaceutically
acceptable salt thereof.
[00661 A compound in the sialic acid biosynthetic pathway or derivative
thereof includes, but
is not limited to, mannosamine, N-acetyl mannosamine (ManNAc), ManNac-6-
phosphate
(ManNAc-6-P), UDP-G1cNA.c, N-acetylneuraminic acid (NeuA.c), NeuAc-9-phosphate
(NeuAc-9-P), sialic acid (i.e., 5-N-acetylneuraminic acid), CMP-sialic acid,
and/or
derivatives thereof or pharmaceutically acceptable salts of the foregoing.
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100671 In some embodiments, the one or more compounds in the sialic acid
biosynthetic
pathway or derivative thereof include N-acetylneuraminic acid (NeuAc) or a
derivative
thereof. Structures of such NettAc or derivatives thereof include, but are not
limited to, those
defined by the formula below:
OR5 0
0Ri
. .
= =
R70 = = =OR2
5R:3 =
. .
F-IN ===
oRs
R4 0 wherein each R1, R7, R3, R5, R6, or R7 is
independently hydrogen, lower alkano2,,,4, carbox2,,,late or lower alkyl; and
R, is lower alkyl,
lower alkanoylalkyl or lower alkyl alkanoyloxy.
[00681 In some embodiments, the one or more compounds in the sialic acid
biosynthetic
pathway or derivative thereof include ManNAc or a derivative thereof
Structures of such
MariN.A.c and derivatives thereof include, but are not limited to, those
defined by the formula
H 0R5
R40 \
R30
Wherein each R1, R3. R4, or R5 is independently hydrogen, lower alkanoyl,
carboxylate or
lower alkyl; and R2 is lower alkyl, lower alkanoylalkyl or lower alkyl
alkano:y4oxy.
[00691 The term lower alkyl refers to (Ct-C6)alkyl. A lower alkyl includes
methyl, ethyl,
propyl, isopropyl, butyl, iso-hutyl, sec-butyl, pentyl, 3-pentyl, .hexyl as
well as (Cy
C6)cycloalkyl moieties (e.g., cyclopropyl, cyclobtql, cyclopentyl, or
cyclohexyl), (C3-
C6)cycloalkyl(Ci-C6)alkyl (e.g., cyclopropylmethyl, cyclobutylmethyl,
cyclopentylmetbyl,
cyclohexylmethyl, 2-cyclopropylethyl, 2-cyclobutylethyl, 2- cycloperlethyl, or
2-
cyclohexylethyl), (CpC6)alkoxy (e.g., methoxy, ethoxy, propoxy, isopropoxy,
butoxy, iso-
butoxy, sec-butoxy, pentoxy, 3-pentoxy, or hexyloxy) (C2-C6)alkenyl (e.g.,
vinyl,
propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1,-pentenyl, 2-
pentenyl, 3-pentenyl, 4-
pentenyt, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, or 5-hexenyl), (C2-
C6)alkynyl (e.g.,
ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butyn.yl, 3-butynyl, 1-pentynyl,
2-pentynyl, 3-
pentynyl, 4-pent:yrnyl, 1 -hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, or 5-
hexynyl), (C1-
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C6)alkanoyl (e.g., acetyl., propanoyi or butanoyl), halo(Ci-C6)alkyl (e.g.,
iodomethyl,
bromometh.yl, chloromethyl, fluoramethyl, trifluoramethyl, 2-chloroeth.yi, 2-
fluoroethyl,
2,2,2-trifluoroethyl, or pentafluoroethyl), hydroxy(Ci-C6)alkyl (e.g.,
hydroxymethyl, 1-
hydroxyethyl, 2-hydroxyethyl, I-hydroxypropyl, 2-hydroxypropyl, 3-
hydroxypropyl, 1-
hydroxy butyl, 4-hydroxybutyl, 1-hydroxypentyl, 5-hydrowentyl, 1-
hydroxyh.exyl, or 6-
hy droxyhexyl), (C1-C6)alkoxycarbonyi (e.g.,
methoxycarbonyl, ethoxycarbony
propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl,
pentoxycarbonyl, or
hexyloxycarbonyl), (C1-C6)alkylthio (e.g., methyithio, ethylthio, propylthio,
isopropylthio,
isobutylthio, pentylthio, or h.exylthio), and/or (C2-C6)alkanoyloxy (e.g.,
acetoxy,
propanoyloxy, butanoyloxy, isobutanoyloxy, pentanoyloxy, or hexanoyloxy).
[00701 in some embodiments, R2 is methyl, and each of RI. R. R. and R5 is
hydrogen. in
some embodiments, the ManNAc or derivative thereof is N-acetyl mannosamine
(ManNAc).
In some embodiments, the ManNAc or derivative thereof is N-
levulinoylmannosamine
(ManLev) or N-azidoacetylmannosamine (MariNAz).
100711 In one variation, the one or more compounds in the sialic acid
biosynthetic pathway
or derivative thereof is an ester of a compound in the sialic acid
biosynthetic pathway. In one
aspect, the one or more compounds in the sialic acid biosynthetic pathway or
derivative
thereof is an ester of sialic acid or MaNAc. in a particular variation, the
one or more
compounds in the sialic acid biosynthetic pathway or derivative thereof is an
ester of sialic
acid. In one aspect, the one or more compounds in the sialic acid biosynthetic
pathway or
derivative thereof is a prodrug of sialic acid. See also WO 2010/131712,
published November
18, 2010, for derivatives of compounds in the sialic acid biosynthetic
pathway, which is
incorporated herein by reference in its entirety and specifically with respect
to compounds
(e.g., derivatives of compounds in the sialic acid biosynthetic pathway)
detailed therein.
[00721 .In one aspect, a derivative of one or more compounds in the sialic
acid biosynthetic
pathway (e.g., a derivative of sialic acid or MaNAc) is an effective substrate
replacement for
sialic acid, such as in an individual who has or is suspected of having a
sialic acid deficiency
disorder. A derivative of one or more compounds in the sialic acid
biosynthetic pathway
(e.g., a derivative of sialic acid or MaNAc), or an. extended release
formulation comprising a
derivative of one or more compounds in the sialic acid biosynthetic pathway
(e.g., a
derivative of sialic acid or MaNA.c) may exhibit any one or more of the
following
characteristics: (i) capable of delivering to an individual in need thereof a
therapeutically
effective amount of one or more compounds in the sia lie acid pathway or
derivatives thereof
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over a period of about or greater than about any of 8, 9, 10, 11, 12, 13, 14,
15, 16, 17, 18, 19,
or 20 hours; (ii) capable of delivering to an individual in need thereof a
substantially constant
therapeutically effective amount of one or more compounds in the sialic acid
pathway or
derivatives thereof over a period of about or greater than about any of 8, 9,
10, 11, 12, 13,
14, 15, 16, 17, 18, 19, or 20 hours; (iii) capable of delivering to an
individual in need thereof
a therapeutically effective amount of one or more compounds in the sialic acid
pathway or
derivatives thereof with a Tmax of between about any of 2-6 hours, 2-5 hours,
or 3-6 hours
during each dosing interval; (iv) capable of delivering to an individual in
need thereof a
therapeutically effective amount of one or more compounds in the sialic acid
pathway or
derivatives thereof with a C. of about 0.1 ¨ 0.9 ggirnL, 0.1-100 ug/mL, 0.2-
0.3 u.g/mL, or
0.5-100 p.g/m1.4 (v) capable of delivering to an individual in need thereof a
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
with a trough level of about 0.05 ¨0.2 lug/mL, 0.05 ¨0.3 tig/mL, 0.1 ¨0.3
1.1.g/mL, or 0.1 ¨20
1.1.g/m1L; (vi) capable of delivering to an individual in need thereof a
therapeutically effective
amount of one or more compounds in the sialic acid pathway or derivatives
thereof with less
than about any of 10%, 20%, 30%, 40%, 50%, 60%, or 70% excreted after one
hour; (vii)
capable of delivering to an individual in need thereof between about any of
0.01-750
mg/kg/day, 0.5-500 mg/kg/day, 1-250 mg/kg/day, 2.5-100 mg/kg/day, or 5-50
mg/kg/day of
one or more compounds in the sialic acid pathway or derivatives thereof or a
pharmaceutically acceptable salt of the foregoing; (viii) capable of
delivering to an individual
in need thereof between about any of 0.01-750 mg/kg/day, 0.5-500 mg/kg/day, 1-
250
mg/kg/day, 2.5-100 mg/kg/day, or 5-50 mg/kg/day of one or more compounds in
the sialic
acid pathway or derivatives thereof or a pharmaceutically acceptable salt of
the foregoing;
(ix) has an absolute bioavailability of about 1 to about 50%; (x) has a
bioavailability based on
sialic acid levels in the urine of about 0.5 to about 100%; and (xi) has a
mean residence time
(MRT) of at least about 3.5 hours.
(0073) In some embodiments, the one or more compounds in the sialic acid
biosynthetic
pathway or derivative thereof include sialic acid or a derivative thereof. In
some
embodiments, the sialic acid or derivative thereof is sialic acid. In some
embodiments, the
sialic acid or derivative thereof is a sialic acid analog such as N-levulinoyl
sialic acid
(SiaLev) or N-azidoacetyl sialic acid (SiaNAz). In some embodiments, the
sialic acid is
bound as a glycoconjugate. In some embodiments, the sialic acid or derivative
thereof is an
unnatural precursor such as sialylactose.
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100741 In some embodiments, the extended release formulation comprises about
any of one,
two, three, or four compounds in the sialic acid biosynthetic pathway or a
derivative thereof.
In some embodiments, the extended release formulation comprises two compounds
in the
sialic acid biosynthetic pathway or a derivative thereof. Therefore, for
example, the extended
release formulation may include ManNAc or a derivative thereof and sialic acid
or a
derivative thereof. More particularly, the extended release formulation may
include ManNAc
and sialic acid.
[00751 In embodiments of any of the extended release formulations, the amount
of one or
more compounds in the sialic acid biosynthetic pathway or derivative thereof
in the extended
release formulation is an amount effective to increase sialic acid production
and/or increase
sialylation (e.g., maximal restoration of sialylation).
[00761 The ratio of the two or more compounds in the sialic acid biosynthetic
pathway or
derivative thereof, in some embodiments, is a ratio which minimizes feedback
inhibition of
the sialic acid biosynthetic pathway. In some embodiments, the ratio of the
two or more
compounds in the sialic acid biosynthetic pathway or derivative thereof is a
ratio which
allows efficient delivery of the two or more compounds in the sialic acid
biosynthetic
pathway or derivative thereof to muscle cells. In some embodiments, the ratio
of the two or
more compounds in the sialic acid biosynthetic pathway or derivative thereof
is a ratio which
minimizes feedback inhibition of the sialic acid biosynthetic pathway and
allows efficient
delivery of the two or more compounds in the sialic acid biosynthetic pathway
or derivative
thereof to muscle cells. In some embodiments, the two or more compounds in the
sialic acid
biosynthetic pathway or derivative there of are ManNAc or a derivative thereof
and sialic
acid or a derivative thereof. For example, in some embodiments, the ratio of
ManNAc and
sialic acid is a ratio which minimizes feedback inhibition of the sialic acid
biosynthetic
pathway and allows efficient delivery of ManNAc and/or sialic acid to muscle
cells. The
combination may optimally spread out the replacement of intermediates,
enhancing optimal
distribution to all cell types with different metabolisms. Methods of testing
restoration of
sialylation and determining the best ratio of the two or more compounds in the
sialic acid
biosynthetic pathway or derivative thereof using in vitro HIBM muscle cells
are known in the
art. See e.g., Noguchi S. et al., .1. Bio. chem. 279(12):11402-7 (2004). This
may involve
evaluating muscle derived proteins for optimal sialylation such as soluble
forms of neural cell
adhesion molecule (NCAM) (Ricci et al., Neurology 66:755-758 (2006),
evaluating sialic
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metabolite or GMP-sialic acid levels in tissue samples, or assessing
sialylated proteins on the
surface of muscle or other cell.s. Noguchi. S. et .Bio. Chem. 279(12):11402-
7 (2004).
[00771 In embodiments in which the extended release fotmulation comprises two
compounds
in the sialic acid biosynthetic pathway or a derivative thereof, the two
compounds in the
extended release formulation may be present in a weight to weight percentage
of between
about any of 5%-95%:95%-5%, 5%-50%:95%-50%, or 10%-40%:90%-60%. The two
compounds in the extended release formulation may be present in a weight to
weight
percentage of about any of 90%:10%, 80%:20%, 70%:30%, 60%:40%, 50%:50%,
40%:60%,
30%:70%, 20%:80%, or 10%:90%. In some embodiments, the two compounds in the
extended release formulation are in a weight to weight percent of about
50%:50%. In some
embodiments, the one or more compounds in the sialic acid pathway or
derivatives thereof
include ManNAc or a derivative thereof and/or sialic acid or a derivative
thereof. Therefore,
for example, the extended release formulation may include ManNAc and sialic
acid wherein
the weight to weight percentage of ManNAc to sialic acid is about any of
90%:10%,
80%:20%, 70%:30%, 60%:40%, 50%:50%, 40%:60%, 30%:70%, 20%:80%, or 10%:90%.
Polymer
[00781 The extended release formulations comprising one or more compounds in
the sialic
acid biosynthetic pathway or a derivative thereof as described herein may
include one or
more polymers. The polymer may be a natural polymer (e.g., polysaccharide or
protein.),
modified natural polymer, and/or synthetic polymer. The polymer may be, for
example, a
hydrophobic polymer, hydrophilic polymer, hydrogel, soluble polymer,
biodegradable
polymer, nonbiodegradable polymer, and/or mucoadhesive polymer.
[0079] In sonic embodiments, the polymer is a hydrophobic polymer. Examples of
hydrophobic polymers include polyethylene, polyvinyl chloride, ethyl cellulose
or acxylato
polymers and their copolymers.
[00801 :In some embodiments, the polymer is a hydrophilic polymer. Examples of
hydrophilic
polymers include a) cellulose derivatives such as methylcellulose (MC),
hydroxyethyl-
cellulose, hydroxypropylmethyl-cellulose (1-1PMC), or sodium
carboxymethylcellulose, b)
noncellulose natural or semisynthetic polymers such as agar-agar, carob gum,
alginates,
molasses, polysaccharides of mannose and galactose, or chitosa.n and modified
starches and
c) polymers of acr2,,,,lic acid such as carbopol polymers.
[0081] In some embodiments, the polymer is a hydrogel. Examples of hydrogels
include, but
are not limited to, polyh.2,,,,droxyethyle meth.2,,,,lacrylate (PHEMA),
polyvinyl alcohol (P-VA),
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polyvinyl pyrrolidone (PVP), polyethylene oxide (PEO), or polyacrylamide (PA).
In some
embodiments, the hydrogel is polyethylene oxide (e.g., PolyoxTM water soluble
resin, Dow
Chemical Company, Mich., USA).
[00821 In some embodiments, the polymer is a soluble polymer. Examples of
soluble
polymers include, but are not limited to, polyethylene glycol (PEG), PVA, PVP,
or HPMC.
[00831 In some embodiments, the polymer is a biodegradable polymer. Examples
of
biodegradable polymers include, but are not limited to, polylactic acid (PLA),
polyglycolic
acid (PGA), poly(lacticiglycolic acid) (PLGA), polycaprolactone (PCL),
polyanhydrides, or
polyorthoesters.
100841 In some embodiments, the polymer is a nonbiodegradable polymer.
Examples of
nonbiodegradable polymers include, but are not limited to, polyethylene vinyl
acetate,
polydimethyl siloxane (PDS), polyether urethane (PEU), polyvinyl chloride
(PVC), cellulose
acetate (CA), or ethyl cellulose (EC).
100851 In some embodiments, the polymer is a mucoadhesive polymer. Examples of
mucoadhesive polymers include, but are not limited to, polycarbophil, sodium
carboxymethyl
cellulose, polyacrylic acid, tragacanth, methyl cellulose, pectin, natural
gums, xanthan gum,
guar gum, or karaya gum.
100861 In some embodiments, the extended release pharmaceutical formulation
includes two
polymers. In some embodiments, the polymer is not polylactide. In some
embodiments, the
polymer is not a polylactide copolymer such as PLGA.
[00871 In some embodiments, the extended release formulation comprises one or
more
polymers selected from the group consisting of a) a water-swellable, pH
independent
polymer, b) a anionic, pH-dependent, gel-forming copolymer, c) a cationic
polymer, and d) a
hydrocolloid polymer. In one variation, the extended release formulation
comprises sialic
acid, or a pharmaceutically acceptable salt thereof, as the therapeutic agent.
In another
variation, the extended release formulation comprises MaNAc, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agent. in another variation, the
extended release
formulation comprises sialic acid, or a pharmaceutically acceptable salt
thereof and MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agents. In
another variation,
the extended release formulation comprises a prodrug of one or more compounds
in the sialic
acid biosynthetic pathway such as a prodrug of sialic acid, or a
pharmaceutically acceptable
salt thereof, as the therapeutic agent. In another variation, the extended
release formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof, as the
therapeutic agent
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and further comprises a hydrocolloid polymer, an anionic, pH-dependent gel
forming co-
polymer and a water swellable, pH independent polymer and optionally further
comprises a
lubricant and/or an excipient. In a particular variation, the extended release
formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof,
carrageenan, sodium
alginate and either hypromellose or polyethylene oxide. In a further
variation, the extended
release formulation comprises sialic acid, or a pharmaceutically acceptable
salt thereof,
carrageenan, sodium alginate, either hypromellose or polyethylene oxide,
magnesium stearate
and rnicrocrystalline cellulose and colloidal silicon dioxide. In one aspect,
the extended
release formulation is a formulation of Table E. In yet another aspect, the
extended release
formulation is a formulation of Table 8. In yet another aspect, the extended
release
formulation is a formulation of Example 6. In yet another aspect, the extended
release
formulation is a formulation of Example 7.
[00881 Examples of a water-swellable, pH independent polymer include, but are
not limited
to, carbohydrate-based polymers such as, for example, hypromellose (formerly
known as the
family of hydroxypropyl methylcellulose), hydroxypropyl ethyl celluloses,
hydroxypropyl
cellulose, hydroxyethyl cellulose, methyl cellulose or other constituents
Grades of these
hypromellose copolymers typically used with the present invention include the
E and K series
such as for example, Dow Chemical Company's (Midland, Mich. USA) or Aqualon's
(with a
North American presence in Wilmington, Del.) E4M, El0M, KlOOLV, K4M, K15M,
K25M,
KIOOM, K200M and mixtures of various molecular weights and grades. Grades of
hydroxyethyl cellulose include, for example, Aqualon's Natrasol polymers HHX
(mol. Wt.
1,300,000), I-EX (mol. wt. 1,000,000), H (mol. wt. 1,000,000), M (mol. wt.
720,000 and G
(mol. wt. 1,150,000), and mixtures thereof. Grades of hydroxypropyl cellulose
include, for
example, Aqualon's HPC polymers MF and MXF (mol. wt. 580,000) and KF and HXF
(mol.
wt. 1,150,000), and mixtures thereof. Grades and ethyl cellulose include, for
example, Dow
Chemical Company's Ethocel polymers 7FP, 10FP and 100FP and Aqualon's polymers
TioEc, N7, N10, N17, N22, N50, N100 and N200, and mixtures thereof. In some
embodiments, the water-swellable, pH independent polymer is hypromellose
(e.g.,
hypromellose Type 2208). In some embodiments, the water-swellable, pH
independent
polymer is Methocel (e.g., Methocel K1OOMPremium CR, Colorcon).
[00891 Examples of anionic, pH-dependent, gel-forming copolymer include, but
are not
limited to, mono-valent alginate salt such as sodium, potassium or ammonium
alginate salts,
or combinations thereof, and sodium carboxymethyl cellulose and the like, or
mixtures of one
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or more alginate salt and carboxymethyl cellulose and the like. In some
embodiments, the
anionic, pH-dependent, gel-forming copolymer is sodium alginate (e.g.,
Protanal , FMC
BioPolymer).
[00901 Examples of a cationic polymer include, for example, chitosan or a
derivative thereof
including, for example, trimethylchitosan and quartermised chitosan, and
chitosan-derived
materials including, for example, those taught in U.S. Pat. No. 5,747,475.
Either high or low
molecular weight chitosan products can be used in the pharmaceutical
formulations of the
present invention and are readily available in pharmaceutical grade from
suppliers located
world-wide.
100911 The hydrocolloid polymer used in the formulations of the present
invention can be
carrageenan. Carrageenans are available as iota, kappa and lambda
carrageenans, with iota
being used most frequently used and lambda being used least frequently.
Various salt forms
of carrageenans are also available including, for example sodium carrageenan.
Typically used
grades of iota carrageenan include, without limitation, carrageenan NF AEP
brand colloids
(Hadley, N.Y. USA) FD433 (1% viscosity; 300-400 cps) and FD384 (1% viscosity;
about
100 cps). Viscosity of other carrageenan products ranges from about 50 to
about 4000 cps. in
some embodiments, the carrageenan is lambda carrageenan (e.g., Viscarin GP-
209, FMC
BioPolymer). In some embodiments, the carrageenan has a viscosity of about
1500-2000 cPs.
In some embodiments, the carrageenan has a viscosity of about 1600 cPs.
[00921 The formulation and polymers useful in the extended release formulation
are further
described in U.S. Patent Application 2010/0160363, published on June 24, 2010,
and U.S.
Patent Application 2010/0159001, published June 24, 2010, which are
incorporated herein by
reference in their entireties and specifically with respect to the polymers
provided therein.
[00931 In some embodiments, the extended release formulation comprises a water-
swellable,
pH independent polymer (e.g., hypromellose). In some embodiments, the extended
release
formulation further comprises an anionic, pH-dependent, gel-forming copolymer
(e.g., an
alginate salt). In some embodiments, the extended release formulation further
comprises a
hydrocolloid polymer (e.g., carrageenan). In some embodiments, the extended
release
formulation comprises a water-swellable, pH independent polymer (e.g.
hypromellose), an
anionic, pH-dependent, gel-forming copolymer (e.g., an alginate salt) and a
hydrocolloid
polymer (e.g., a carrageenan). In some embodiments, the extended release
formulation
comprises hypromellose (e.g. hypromellose Type 2208 or Methocel K1 00M),
sodium
alginate (e.g. Protanal) and a lambda carrageenan (e.g. Viscarin GP-209). In
one variation,
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the extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agent. In another variation, the extended release
formulation
comprises MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agent. In
another variation, the extended release formulation comprises sialic acid, or
a
pharmaceutically acceptable salt thereof and MaNAc, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agents. In another variation, the extended release
formulation
comprises a prodrug of one or more compounds in the sialic acid biosynthetic
pathway such
as a prodrug of sialic acid, or a pharmaceutically acceptable salt thereof, as
the therapeutic
agent. In another variation, the extended release formulation comprises sialic
acid, or a
pharmaceutically acceptable salt thereof, as the therapeutic agent and further
comprises a
hydrocolloid polymer, an anionic, pH-dependent gel forming co-polymer and a
water
swellable, pH independent polymer and optionally further comprises a lubricant
and/or an
excipient. In a particular variation, the extended release formulation
comprises sialic acid, or
a pharmaceutically acceptable salt thereof, carrageenan, sodium alginate and
either
hypromellose or polyethylene oxide. In a further variation, the extended
release formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof,
carrageenan, sodium
alginate, either hypromellose or polyethylene oxide, magnesium stearate and
microctystalline
cellulose and colloidal silicon dioxide. In one aspect, the extended release
formulation is a
formulation of Table E. In yet another aspect, the extended release
formulation is a
formulation of Table 8. In yet another aspect, the extended release
formulation is a
formulation of Example 6. In yet another aspect, the extended release
formulation is a
formulation of Example 7.
10094) In some embodiments, the extended release formulation comprises a
hydrogel (e.g., a
polyethylene oxide). In some embodiments, the extended release formulation
further
comprises an anionic, pH-dependent, gel-forming copolymer (e.g. an alginate
salt). In some
embodiments, the extended release formulation further comprises a hydrocolloid
polymer
(e.g., carrageenan). In some embodiments, the extended release formulation
comprises a
hydrogel (e.g., a polyethylene oxide), an anionic, pH-dependent, gel-forming
copolymer
(e.g., an alginate salt) and a hydrocolloid polymer (e.g., a carrageenan). In
some
embodiments, the extended release formulation comprises polyethylene oxide
(e.g. Polyox
WSR), sodium alginate (e.g. Protanal) and a lambda carrageenan (e.g. Viscarin
GP-209). In
one variation, the extended release formulation comprises sialic acid, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agent. In another variation, the
extended release
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formulation comprises MaNAc, or a pharmaceutically acceptable salt thereof, as
the
therapeutic agent. In another variation, the extended release formulation
comprises sialic
acid, or a pharmaceutically acceptable salt thereof and MaNAc, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agents. In another variation, the
extended release
formulation comprises a prodrug of one or more compounds in the sialic acid
biosynthetic
pathway such as a prodrug of sialic acid, or a pharmaceutically acceptable
salt thereof, as the
therapeutic agent. in another variation, the extended release formulation
comprises sialic
acid, or a pharmaceutically acceptable salt thereof, as the therapeutic agent
and further
comprises a hydrocolloid polymer, an anionic, pH-dependent gel forming co-
polymer and a
water swellable, pH independent polymer and optionally further comprises a
lubricant and/or
an excipient. in a particular variation, the extended release formulation
comprises sialic acid,
or a pharmaceutically acceptable salt thereof, carrageenan, sodium alginate
and either
hypromellose or polyethylene oxide. In a further variation, the extended
release formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof,
carrageenan, sodium
alginate, either hypromellose or polyethylene oxide, magnesium stearate and
microcrystalline
cellulose and colloidal silicon dioxide. In one aspect, the extended release
formulation is a
formulation of Table E. In yet another aspect, the extended release
formulation is a
formulation of Table 8. In yet another aspect, the extended release
formulation is a
formulation of Example 6. In yet another aspect, the extended release
formulation is a
formulation of Example 7.
100951 In one variation, the extended release formulation comprises: (i) a
hydrocolloid
polymer; (ii) an anionic, pH-dependent , gel forming co-polymer, and (iii)
either a water-
swellable, pH independent polymer or a hydrogel. In one variation, the
extended release
formulation comprises sialic acid, or a pharmaceutically acceptable salt
thereof, as the
therapeutic agent. In another variation, the extended release formulation
comprises MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agent. In
another variation,
the extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof and MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agents.
in another variation, the extended release formulation comprises a prodrug of
one or more
compounds in the sialic acid biosynthetic pathway such as a prodrug of sialic
acid, or a
pharmaceutically acceptable salt thereof, as the therapeutic agent. In another
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agent and further comprises a hydrocolloid
polymer, an anionic,
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pH-dependent gel forming co-polymer and a water swellable, pH independent
polymer and
optionally further comprises a lubricant and/or an excipient. In a particular
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, carrageenan, sodium alginate and either hypromellose or polyethylene
oxide. In a
further variation, the extended release formulation comprises sialic acid, or
a
pharmaceutically acceptable salt thereof, carrageenan, sodium alginate, either
hypromellose
or polyethylene oxide, magnesium stearate and microcrystalline cellulose and
colloidal
silicon dioxide. In one aspect, the extended release formulation is a
formulation of Table E.
In yet another aspect, the extended release formulation is a formulation of
Table 8. In yet
another aspect, the extended release formulation is a formulation of Example
6. In yet
another aspect, the extended release formulation is a formulation of Example
7. An extended
release formulation in one variation comprises a therapeutic agent as detailed
herein (e.g.,
sialic acid) and (i) a hydrocolloid polymer; (ii) an anionic, pH-dependent,
gel forming co-
polymer, and (iii) either a water-swellable, pH independent polymer or a
hydrogel.
Exemplary extended release formulations include those listed in Table A, where
it is
understood that an extended release formulation may comprise any of the listed
therapeutic
agents in combination with at least one of any of polymers 1, 2, 3A or 3B the
same as if each
and every combination of therapeutic agent and polymer or combination of
polymers were
specifically and individually listed. Although particular formulations may
comprise a
therapeutic agent of Table A and any one or more of a polymer selected from
Polymers 1, 2
and 3 (A and/or B) of Table A, in a particular variation, an extended release
formulation
comprises a therapeutic agent of Table A, a polymer 1 of Table A, a polymer 2
of Table A
and either a polymer 3A of Table A or a polymer 3B of Table A the same as if
each and
every combination of therapeutic agent and polymer combination were
specifically and
individually listed. For example, it is understood that in one aspect, an
extended release
formulation comprises sialic acid, carrageenan (e.g., a lambda carrageenan
such as Viscarin
GP-209), an alginate salt (e.g., sodium alginate such as Protanal LF 120M),
and either (i)
hypromellose (e.g., hypromellose Type 2208) or (ii) polyethylene oxide (e.g.,
Polyox), or a
pharmaceutically acceptable salt of any of the foregoing.
Table A. Exemplary Components for use in Extended Release Formulations.
Formulation Component Weight Percent of Component in Formulation
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Formulation Component Weight Percent of Component in Formulation
Therapeutic Agent mannosamine, N-acetyl mannosamine (ManNA.c), ManNac-6-
(A compound in the sialic phosphate (ManNAc-6-P), UDP-GleNAc, N-
acetylneuraminic
acid biosynthetic pathway acid (NeuAc), NeuAc-9-phosphate (NeuAc-9-P), sialic
acid
or derivative thereof or (i.e., 5-N-acetylneuraminic acid), CMP-sialic
acid, andlor
salt of any of the derivatives thereof or pharmaceutically acceptable salts
of the
foregoing) foregoing.
Polymer I Carrageenan (e.g., iota, kappa or lambda carrageenan, or
a salt
(Hydrocolloid Polymer) thereof, such as Viscarin GP-209, FMC BioPolymer)
Polymer 2 Alginate or salt thereof (e.g., sodium, potassium. or
ammonium
(Anionic, pH-dependent, alginate salt such as Protanal , FMC BioPolymer),
gel forming polymer) carboxym.ethyl cellulose or salt thereof (e.g., sodium
carboxymethyl cellulose).
Polymer 3A (Water- Hyprotnellose (e.g., hyprorneliose Type 2208, E and K
series
swellable, pH independent such as for example, Dow Chemical Company's
(Midland,
polymer) Mich. USA) or Aqualon's (with a North American presence
in
Wilmington, Del.) E4M, El0M, KlOOLV, K4M, K15M, K25M,
KlOOM, K200M and mixtures of various molecular weights and
grades); hydroxypropyl ethyl cellulose (Ethocel polymers 7FP,
10FP and 100FP and Aqualon's polymers T1OEC, N7, NIO,
N17, N22, N50, N100 and N200, and mixtures thereof),
hydroxypropyl. cellulose (A.qualon.'s TIPC polymers MF and
MXF (mol. wt. 580,000) and KF and HXF (mol. wt. 1,150,000),
and mixtures thereof); hydroxyeth.yl cellulose (e.g., Aqualon's
Natrasol polymers MIX (m.ol. Wt. 1,300,000), H). (mol. wt.
1,000,000), H (mol. wt. 1,000,000), M (mol. wt. 720,000 and G
(mol. wt. 1,150,000), and mixtures thereof); methyl cellulose..
Polymer 3B (Hydrogel- Polyh.ydroxyethyle meth.ylacrylate (PHEMA), polyvinyl
alcohol
forming polymer) (PVA), polyvinyl pyrrolidone (PVP), polyethylene oxide
(PEO),
polyacrylamide (PA), polyethylene oxide (e.g., PolyoxTM water
soluble resin, Dow Chemical Company, Mich., USA).
[00961 In one variation, an extended release formulation comprises a
therapeutic agent of
Table A, a polymer 1 of Table A, a polymer 2 of Table A and either a polymer
3A or a
polymer 3B of Table A, wherein the composition comprises the therapeutic agent
and
polymers in any one of the weight percent ranges depicted in Table B.
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Table B. Exemplary Weight Percent of Certain Components for Use in Extended
Release
Formulations.
Formulation Component Weight Percent er)f Component in Formulation
Therapeutic Agent (A From about 20 to about 80; from about 20 to about 60;
from
compound in the sialic about 20 to about 50; from about 20 to about 40;
from about 20
acid biosynthetic pathway to about 30; from about 15 to about 60; from about
15 to about
or derivative thereof or 50; from about 15 to about 40; from about 25 to
about 60; from
salt of any of the about 25 to about 50; from 25 to about 40; from about
25 to
foregoing) about 30; from about 30 to about 60; from about 30 to
about 50;
from about 30 to about 45; from about 30 to about 40; from
about 35 to about 60; from about 35 to about 50; from about 35
to about 45; from about 40 to about 45; about any of 35, 36, 37,
38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 and 50.
Polymer 1. (Hydrocolloid From about Ito about 10; from about Ito about 5; from
about 3
Polymer) to about 8; from about 4 to about 6; about any of 1, 2,
3, 4, 5, 6,
7, 8, 9 and 10.
Polymer 2 (Anionic, pH- From about 15 to about 30; from about 15 to about 25;
from
dependent gel forming about 15 to about 20; from about 20 to about 30; from
about 20
polymer) to about 25; from about 2o to about 23; about any of
15, 16, 17,
18, 19, 20, 21, 22, 23, 24 and 25.
Polymer 3A. (Water- From about 20 to about 50; from about 20 to about 40,
from
&wettable, pH independent about 20 to about 30; from about 20 to about 25;
from about 25
polymer) to about 30; from about 22 to about 27; about any of
20, 21, 22,
Or 23, 24, 25, 26, 27, 28, 29 and 30.
Polymer 3B. (Hydrogel-
forming.polymer)
[00971 in another variation, the extended release formulation comprises a
therapeutic
agent (a compound in the sialic acid biosynthetic pathway or derivative
thereof or salt of any
of the foregoing, such as any of the compounds detailed herein, including in
Table A) and a
polymer; wherein the polymer comprises: (i) a hydrocolloid polymer; (ii) an
anionic, pH-
dependent , gel fbrming co-polymer, and (iii) either a water-swellable, pH
independent
polymer or a hydrogel, and wherein the weight percent ratio of polymers
(i):(ii):(iii) is about
1:5:5 or about 1:5:6.
[0098] The combination of (i) a hydrocolloid polymer; (ii) an anionic, pH-
dependent ,
gel forming co-polymer, and (iii) either a water-sweliable, pH independent
polymer or a
hydrogel is believed to provide a unique combination that is particularly
advantageous for the
preparation of oral dosage forms in that the combination results in any one or
more of the
following features: (i) provides a robust formulation (e.g., for tablet
formulation); (i) is pH
independent; and (iii) lends itself to granulation without affecting
dissolution profile.
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[0099] Further descriptions of extended release formulations and
formulation
components are found throughout and below.
[00100] It is understood that reference to relative weight percentages
assumes that the
combined total weight percentages of all components in the formulation add up
to 100. It is
further understood that relative weight percentages of one or more components
may be
adjusted upwards or downwards such that the weight percent of the components
in the
composition combine to a total of 100. In one aspect, the weight percentages
detailed herein
refer to the weight percentages of a formulation blend (e.g., prior to
formulation into a unit
dosage amount such as a tablet, which may be further modified, e.g., by the
addition of a
tablet coating). In another aspect, the weight percentages detailed herein
refer to the weight
percentages of a unit dosage of a formulation, in which the formulation is in
a form. and/or
packaged for administration to an individual (e.g., a tablet that has a
coating).
[00101] The polymer may be present in the extended release formulation in
an amount
ranging from 5 to 40 parts by weight, from. 10 to 20 parts by weight, relative
to 100 parts by
weight of the one or more compounds in the sialic acid pathway or derivatives
thereof. In
some embodiments, the one or more compounds in the sialic acid pathway or
derivatives
thereof in such formulations includes from about 0.1 to 99.9% by weight of the
formulation.
In some embodiments, the one or more compounds in the sialic acid pathway or
derivatives
thereof in such formulations includes about any of between 20%-30%, 30%-40%,
40%-50%,
or 20%-50%. In som.e embodiments, the extend release formulation includes
about any of
between 40%-50%, 50%-60%, 60%-70%, or 50% to 70% by weight of polymer.
[00102] In some embodiments, the drug load of the one or more compounds in
the sialic
acid pathway or derivatives thereof in the extended release formulation
comprises about 20%
to 80% w/w. In some embodiments, the drug load of the one or more compounds in
the sialic
acid pathway or derivatives thereof in the extended release formulation
comprises about any
one of 20% to 60% w/w, 20% - 50% w/w, 20% - 40% w/w, 15% - 60% w/w, 15% - 50%
w/w, 15% - 40% w/w, 25% - .60% w/w, 25% - 50% w/w, 25% - 40% w/w, 30% - 60%
w/w,
30% - 50% w/w, 30% - 45% w/w, 35% - 60% w/w, 35% - 50% w/w, or 35% - 45% w/w.
In
some embodiments, the drug load of the one or more compounds in the sialic
acid pathway or
derivatives thereof in the extended release formulation comprises at least
about any one of
25% wlw, 30% w/w, 35% w/w, 40% w/w, 45% wlw, or 50% w/w. In some embodiments,
the
drug load of the one or more compounds in the sialic acid pathway or
derivatives thereof in
the extended release formulation comprises about 33% w/w. In some embodiments,
the drug
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load of the one or more compounds in the sialic acid pathway or derivatives
thereof in the
extended release formulation comprises about 43% w/w
1001031 In some embodiments, the extended release formulation comprises
about 20 to
about 50 or about 20 to about 40 or about 20 to about 30% w/w of a water-
swellable, pH
independent polymer (e.g., hyprom.ellose). In som.e embodiments, the extended
release
formulation comprises about 25% w/w of a water-swellable, pH independent
polymer (e.g.,
hypromellose).11n some embodiments, the extended release formulation further
comprises
about 20-25% w/w of an anionic, pH-dependent, gel-forming copolymer (e.g., an
alginate
salt). In some embodiments, the extended release formulation further comprises
about
21%w/w of an anionic, pH-dependent, gel-forming copolymer (e.g., an alginate
salt). In some
embodiments, the extended release formulation further comprises about 1-5% w/w
of a
hydrocolloid polymer (e.g., carrageenan). In some embodiments, the extended
release
formulation further comprises about 4% w/w of a hydrocolloid polymer (e.g.,
carrageenan).
In some embodiments, the extended release formulation comprises about 20-30%
w/w of a
water-swellable, pH independent polymer (e.g. hypromellose), about 20-25% w/w
of an
anionic, pH-dependent, gel-forming copolymer (e.g., an alginate salt) and
about 1-5 w/w
of a hydrocolloid polymer (e.g., a carrageenan). In some embodiments, the
extended release
formulation comprises about 20-30% w/w hypromellose (e.g. hypromellose Type
2208 or
Methocel K100M), about 20-25% w/w sodium alginate (e.g. Protanal) and about 1-
5% w/w
lambda carrageenan (e.g. Viscarin GP-209). In some embodiments, the extended
release
formulation comprises about 25% w/w of a water-swellable, pH independent
polymer (e.g.
hypromellose), about 21% w/w of an anionic, pH-dependent, gel-forming
copolym.er (e.g., an
alginate salt) and about 4% w/w of a hydrocolloid polymer (e.g., a
carrageenan). In some
embodiments, the extended release formulation comprises about 25% w/w
hypromellose (e.g.
hypromellose Type 2208 or Methocel K100M), about 21% w/w sodium alginate (e.g.
Protanal) and about 4% w/w lambda carrageenan (e.g. Viscarin GP-209).
[00104] In some embodiments, the extended release formulation comprises
about 20 to
about 50 or about 20 to about 40 or about 20 to about 20-30% w/w of a hydrogel
(e.g., a
polyethylene oxide, Polyox WSR). In some embodiments, the extended release
formulation
comprises about 25% wlw of a hydrogel (e.g., a polyethylene oxide, Polyox
WSR). In some
embodiments, the extended release formulation further comprises about 20-25%
w/w of an
anionic, pH-dependent, gel-forming copolymer (e.g., an alginate salt). In some
embodiments,
the extended release formulation further comprises about 21% w/w of an
anionic, pH-
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dependent, gel-forming copolymer (e.g., an alginate salt). In some
embodiments, the
extended release formulation further comprises about 1-5% w/w of a
hydrocolloid polymer
(e.g., carrageenan). In some embodiments, the extended release formulation
further
comprises about 4% w/w of a hydrocolloid polymer (e.g., carrageenan). In some
embodiments, the extended release formulation comprises about 20-30% w/w of a
hydrogel
(e.g., a polyethylene oxide), about 20-25% w/w of an anionic, pH-dependent,
gel-forming
copolymer (e.g., an alginate salt) and about 1-5% w/w of a hydrocolloid
polymer (e.g., a
carrageenan). In some embodiments, the extended release formulation comprises
about 20-
30% w/w polyethylene oxide (e.g. Polyox WSR), about 20-25% w/w sodium alginate
(e.g.
Protanal) and about 1-5% wlw lambda carrageenan (e.g. Viscarin GP-209). In
some
embodiments, the extended release formulation comprises about 25% w/w of a
hydrogel
(e.g., a polyethylene oxide), about 21% w/w of an anionic, pH-dependent, gel-
forming
copolymer (e.g., an alginate salt) and about 4% w/w of a hydrocolloid polymer
(e.g., a
carrageenan). In some embodiments, the extended release formulation comprises
about 25%
w/w polyethylene oxide (e.g. Polyox WSR), about 21% w/w sodium alginate (e.g.
Protanal)
and about 4% w/w lambda carrageenan (e.g. Viscarin GP-209).
Additional Formulation Components
[00105] The extended release pharmaceutical formulations comprising one or
more
compounds in the sialic acid biosynthetic pathway or derivative thereof as
described herein
may further comprise a diluent, an excipient, an antioxidant, a lubricant, a
colorant, a binder,
a disintegrant, and the like. It is understood that any of the extended
release formulations
detailed herein, including but not limited to those listed under the heading
"Extended Release
Formulations" (e.g., any formulation of Tables A or B) may further comprise a
diluent, an
excipient, an antioxidant, a lubricant, a colorant, a binder, a disintegrant,
and the like as
detailed herein the same as if each and every extended release formulation
further comprising
such a component were specifically and individually listed. In one variation,
the extended
release formulation comprises sialic acid, or a pharmaceutically acceptable
salt thereof, as the
therapeutic agent. In another variation, the extended release formulation
comprises MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agent. In
another variation,
the extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof and MaNAc, or a pharmaceutically acceptable salt thereat as the
therapeutic agents.
In another variation, the extended release formulation comprises a prodrug of
one or more
compounds in the sialic acid biosynthetic pathway such as a prodrug of sialic
acid, or a
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pharmaceutically acceptable salt thereof, as the therapeutic agent. In another
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agent and further comprises a hydrocolloid
polymer, an anionic,
pH-dependent gel forming co-polymer and a water swellable, pH independent
polymer and
optionally further comprises a lubricant and/or an excipient. In a particular
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, carrageenan, sodium alginate and either hypromellose or polyethylene
oxide. in a
further variation, the extended release formulation comprises sialic acid, or
a
pharmaceutically acceptable salt thereof, carrageenan, sodium alginate, either
hypromellose
or polyethylene oxide, magnesium stearate and microcrystalline cellulose and
colloidal
silicon dioxide. In one aspect, the extended release formulation is a
formulation of Table E.
In yet another aspect, the extended release formulation is a formulation of
Table 8. In yet
another aspect, the extended release formulation is a formulation of Example
6. In yet
another aspect, the extended release formulation is a formulation of Example
7.
[00106] The diluent is selected so as not to affect the biological activity of
the
combination. Examples of such diluents are distilled water, buffered water,
physiological
saline, PBS, Ringer's solution, dextrose solution, and Hank's solution. The
pharmaceutical
formulations can also include additional substances to approximate
physiological conditions,
such as pH adjusting and buffering agents, toxicity adjusting agents, wetting
agents and
detergents. The pharmaceutical formulations can also include any of a variety
of stabilizing
agents. Further guidance regarding pharmaceutical formulations that are
suitable for various
types of administration can be found in Remington: The Science and Practice of
Pharmacy,
20th Edition. Baltimore, MD: Lippincott Williams & Wilkins, 2000.
[00107] The excipient may be selected from the group consisting of lactose,
microcrystalline cellulose, corn starch, potato starch, wheat starch, sucrose,
D-manni.tol,
precipitated calcium carbonate, dextrin, pre-gelatinized starch, and
combinations thereof. The
excipient, if present, may be contained in an amount of about 10 to about 90
parts by weight
based on the total weight of the tablet. In some embodiments, the extend
release formulation
includes about any of between 40%-50%, 50%-60%, 60%-70%, or 50% to 70% by
weight of
excipient. In some embodiments, the excipient is microcrystalline cellulose.
In some
embodiments, the excipient is microcrystalline cellulose and colloidal silicon
dioxide (e.g.,
ProSolve SMCC HD90). In some embodiments, the extended release formulation
comprises
about 1-10% w/w of microcrystalline cellulose and colloidal silicon dioxide
(e.g., ProSolv*)
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SMCC HD90). In some embodiments, the extended release formulation comprises
about 5%
w/w of microcrystalline cellulose and colloidal silicon dioxide (e.g., ProSolv
SMCC
HD90).
[00108] The binder may be selected from the group consisting of
hydroxypropylcellulose,
direct tabletted microcrystalline cellulose, HPMC, MC, hydroxyethy I cel I u
lose,
hydroxymethylcellulose, carboxymethyl cellulose, and other cellulose
derivative, PVP, PVA,
paste, arabic gum., dextrin, gelatin, alginates, and combinations thereof. The
binder, if
present, may be used in an amount of about 2 to about 60 parts by weight based
on the total
weight of the tablet.
[00109] The disintegrant may be selected from the group consisting of sodium
starch
gl.ycolate, crosspovidone, cross carmel lose sodium, low-substituted
hydroxypropylcellulose,
starch, carboxymethylcellulose calcium, calcium carbonate, sodium bicarbonate,
and
combinations thereof. The disintegrant, if present, may be contained in an
amount of about
0.1 to about 32 parts by weight based on the total weight of the tablet
composition.
[00110] The lubricant may be selected from the group consisting of magnesium
stearate,
calcium. stearate, talc, light anhydrous sili.cic acid, and solid polyethyl
glycols and
combinations thereof. The lubricant, if present, may be contained in an amount
of about 0.1
to about 20 parts by weight based on the total weight of the tablet. In some
embodiments, the
lubricant is magnesium stearate (e.g., HyQual0). In some embodiments, the
extended release
formulation comprises about 0.1-1%w/w magnesium. stearate (e.g., HyQuale). In
some
embodiments, the extended release formulation comprises about 0.5% w/w
magnesium
stearate (e.g., HyQuale).
[00111] For the colorant, at least one species which can be selected from
titanium dioxide,
iron oxide, magnesium carbonate, calcium sulfate, magnesium oxide, magnesium
hydroxide,
aluminum lakes, for example, Blue No. 1 Aluminum Lake, Red No. 40 Aluminum
Lake, and
the like can be contained in the tablet.
[00112] in another variation, an extended release formulation detailed herein
(including
but not limited to those listed under the heading "Extended Release
Formulations" (e.g., any
formulation of Tables A and B) further comprises an excipient. In a particular
variation, the
excipient comprises microcrystalline cellulose. In a further variation, the
excipient comprises
microcrystalline cellulose and colloidal silicon dioxide. In any such
variations, the extended
release formulation further comprising an excipient (e.g., an excipient
comprising
microcrystalline cellulose and colloidal silicon dioxide) comprises the
excipient in about 1 to
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about 20 or about 1 to about 15 or about 1 to about 10 or about 1 to about 5
or about 5 to
about 20 or about 5 to about 15 or about 5 to about 10 or about any one of 1,
2, 3, 4, 5, 6, 7, 8,
9 or 10 weight percent. In one variation, the extended release formulation
comprises sialic
acid, or a pharmaceutically acceptable salt thereof, as the therapeutic agent.
In another
variation, the extended release formulation comprises MaNAc, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agent. In another variation, the
extended release
formulation comprises sialic acid, or a pharmaceutically acceptable salt
thereof and MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agents. In
another variation,
the extended release formulation comprises a prodrug of one or more compounds
in the sialic
acid biosynthetic pathway such as a prodrug of sialic acid, or a
pharmaceutically acceptable
salt thereof, as the therapeutic agent. In another variation, the extended
release formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof, as the
therapeutic agent
and further comprises a hydrocolloid polymer, an anionic, pH-dependent gel
forming co-
polymer and a water swellable, pH independent polymer and optionally further
comprises a
lubricant and/or an excipient. In a particular variation, the extended release
formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof,
carrageenan, sodium
alginate and either hypromellose or polyethylene oxide. In a further
variation, the extended
release formulation comprises sialic acid, or a pharmaceutically acceptable
salt thereof,
carrageenan, sodium alginate, either hypromellose or polyethylene oxide,
magnesium stearate
and microcrystalline cellulose and colloidal silicon dioxide. In one aspect,
the extended
release formulation is a formulation of Table E. In yet another aspect, the
extended release
formulation is a formulation of Table 8. In yet another aspect, the extended
release
formulation is a formulation of Example 6. In yet another aspect, the extended
release
formulation is a formulation of Example 7.
[00113.1 In another variation, an extended release formulation detailed herein
(including
but not limited to those listed under the heading "Extended Release
Formulations" (e.g., any
formulation of Tables A and B) further comprises a lubricant. In a particular
variation, the
lubricant comprises a stearate salt, such as magnesium stearate. In any such
variations, the
extended release formulation further comprising a lubricant (e.g., a stearate
salt such as
magnesium stearate) comprises the lubricant in about 0.1 to about 2 or about
0.1 to about 1.5
or about 0.1 to about 1.0 or about 0.01 to about 0.09-5 or about 0.1 to about
0.8 or about 0.1
to about 0.7 or about 0.1 to about 0.6 or about 0.1 to about 0.5 or about 0.2
to about 0.8 or
about 0.3 to about 0.7 or about 0.4 to about 0.6 or about any one of 0.1, 0.2,
0.3, 0.4, 0.5, 0.6,
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0.7, 0.8, 0.9 or 1.0 weight percent. In one variation, the extended release
formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof, as the
therapeutic agent.
In another variation, the extended release formulation comprises MaNAc, or a
pharmaceutically acceptable salt thereof, as the therapeutic agent. In another
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof and MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agents.
In another variation, the extended release formulation comprises a prodrug of
one or more
compounds in the sialic acid biosynthetic pathway such as a prodtug of sialic
acid, or a
pharmaceutically acceptable salt thereof, as the therapeutic agent. In another
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agent and further comprises a hydrocolloid
polymer, an anionic,
pH-dependent gel forming co-polymer and a water swellable, pH independent
polymer and
optionally further comprises a lubricant and/or an excipient. In a particular
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, carrageenan, sodium alginate and either hypromellose or polyethylene
oxide. In a
further variation, the extended release formulation comprises sialic acid, or
a
pharmaceutically acceptable salt thereof, carrageenan, sodium alginate, either
hypromellose
or polyethylene oxide, magnesium stearate and microcrystalline cellulose and
colloidal
silicon dioxide. In one aspect, the extended release formulation is a
formulation of Table E.
In yet another aspect, the extended release formulation is a formulation of
Table 8. In yet
another aspect, the extended release formulation is a formulation of Example
6. In yet
another aspect, the extended release formulation is a formulation of Example
7.
[00114] In a further variation, an extended release formulation detailed
herein (including
but not limited to those listed under the heading "Extended Release
Formulations" (e.g., any
formulation of Tables A and B) further comprises both an excipient and a
lubricant. In any
such variation, the formulation further comprising both an excipient and a
lubricant
comprises the excipient (e.g., an excipient comprising microaystalline
cellulose and colloidal
silicon dioxide) in about 1 to about 20 or about 1 to about 15 or about 1 to
about 10 or about
1 to about 5 or about 5 to about 20 or about 5 to about 15 or about 5 to about
10 or about any
one of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 weight percent and comprises the
lubricant (e.g., a stearate
salt such as magnesium stearate) in about 0.1 to about 2 or about 0.1 to about
1.5 or about 0.1
to about 1.0 or about 0.01 to about 0.09-5 or about 0.1 to about 0.8 or about
0.1 to about 0.7
or about 0.1 to about 0.6 or about 0.1 to about 0.5 or about 0.2 to about 0.8
or about 0.3 to
31
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about 0.7 or about 0.4 to about 0.6 or about any one of 0.1, 0.2, 0.3, 0.4,
0.5, 0.6, 0.7, 0.8, 0.9
or 1.0 weight percent. In a further variation, the formulation further
comprising both an
excipient and a lubricant comprises the excipient (e.g., an excipient
comprising
microcrystalline cellulose and colloidal silicon dioxide) in a weight percent
ratio to lubricant
(e.g., a stearate salt such as magnesium stearate) of about any of 1:10 or
1:11 or 1: 9 or
1:10.5. In one variation, the extended release formulation comprises sialic
acid, or a
pharmaceutically acceptable salt thereof, as the therapeutic agent. In another
variation, the
extended release formulation comprises MaNAc, or a pharmaceutically acceptable
salt
thereof, as the therapeutic agent. In another variation, the extended release
formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof and
MaNAc, or a
pharmaceutically acceptable salt thereof, as the therapeutic agents. In
another variation, the
extended release formulation comprises a prodrug of one or more compounds in
the sialic
acid biosynthetic pathway such as a prodrug of sialic acid, or a
pharmaceutically acceptable
salt thereof, as the therapeutic agent. In another variation, the extended
release formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof, as the
therapeutic agent
and further comprises a hydrocoll.oid polymer, an anionic, pH-dependent gel
forming co-
polymer and a water swellable, pH independent polymer and optionally further
comprises a
lubricant and/or an excipient. In a particular variation, the extended release
formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof,
carrageenan, sodium
alginate and either hypromellose or polyethylene oxide. In a further
variation, the extended
release formulation comprises sialic acid, or a pharmaceutically acceptable
salt thereof,
carrageen.an., sodium alginate, either hypromellose or polyethylene oxide,
magnesium. stearate
and microcrystalline cellulose and colloidal silicon dioxide. In one aspect,
the extended
release formulation is a formulation of Table E. In yet another aspect, the
extended release
formulation is a formulation of Table 8. In yet another aspect, the extended
release
formulation is a formulation of Example 6. In yet another aspect, the extended
release
formulation is a formulation of Example 7.
[00115] Particular extended release formulations include those listed in Table
C, where the
compositions comprise a therapeutic agent, a polymer 1, a polymer 2, either a
polymer 3A or
a polymer 3B, an excipient and a lubricant, and it is understood that each and
every
combination of such components is intended the same as if each and every
combination were
specifically and individually listed.
Table C. Exemplary Extended Release Formulation Compositions.
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Formulation Component Exemplaty Specific Components
Therapeutic Agent mannosamine, N-acetyl. mannosamine (ManNAc), ManNac-6-
(A compound in the phosphate (ManNAc-6-P), UDP-GIcNAc, N-acetylneuraminic
sialic acid biosynthetic acid (NeuAc), NeuAc-9-phosphate (NeuAc-9-P),
si.alic acid (i.e.,
pathway or derivative 5-N-acetylneuraminic acid), CMP-sialic acid, and/or
derivatives
thereof or salt of any of thereof or pharmaceutically acceptable salts of the
foregoing.
the foregoing)
Polymer I Carrageen.an (e.g., iota, kappa or lambda carrageenan, or
a salt
(Hydrocolloid Polymer) thereof, such as Viscarin GP-209, FMC BioPolyiner)
Polymer 2 Alginate or salt thereof (e.g., sodium, potassium or
ammonium
(Anionic, pH-dependent alginate salt such as Protanali1D, FMC BioPolymer),
, gel forming polymer) carboxymethyl cellulose or salt thereof (e.g.,
sodium.
carboxymethyl cellulose).
Polymer 3A (Water- Hypromellose (e.g., hypromellose Type 2208, E and K
series
swellable, pH such as for example, Dow Chemical Company's (Midland,
Mich.
independent polymer) USA) or Aqualon's (with a North American presence in
Wilmington, Del.) E4M, E1OM, KlOOLV, K4M, Kl5M, K25M,
K1 00M, K200M and mixtures of various molecular weights and
grades); hydroxypropyl ethyl cellulose (Ethocel polymers 7FP,
10FP and 100FP and Aqualon's polymers TiOEC, N7, N10, N17,
N22, N50, N1.00 and N200, and mixtures thereof), hydroxypropyl
cellulose (.Aqualon's FIPC polymers MF and MXF (mol. wt.
580,000) and KF and HXF (mol. wt. 1,150,000), and mixtures
thereof); h.ydroxyethyl cellulose (e.g., Aqualon's Natrasol
polymers HHX (mol. Wt. 1,300,000), HX (mol. wt. 1,000,000), H
(mol. wt. 1,000,000), M (mol. wt. 720,000 and G (mol. wt.
1,150,000), and mixtures thereof); methyl cellulose..
Polymer 3B (Hydrogel- Polyhydroxyethyle methylacrylate (PI-HEMA), polyvinyl
alcohol
forming polymer) (PVA), polyvinyl pyrrolidone (PVP), polyethylene oxide
(PEO),
polyacrylamide (PA), polyethylene oxide (e.g., PolyoxTM water
soluble resin, Dow Chemical Company, Mich., USA).
Excipient lactose, microcrystalline cellulose, corn starch, potato
starch,
wheat starch, sucrose, D-mannitol, precipitated calcium.
carbonate, dextrin, pre-gelatinized starch, microcrystalline
cellulose and colloidal silicon dioxide (e.g., ProSolVE SMCC
HD90) and combinations thereof.
Lubricant Stearate salt (such as magnesium stearate (e.g., HyQuale)
and
calcium stearate), talc, light anhydrous silicic acid, and solid
polyeth.y1 glycol.s and combinations thereof
[00116] In one variation, an extended release formulation is a composition as
detailed in
Table C, wherein the composition comprises the formulation components in any
one of the
weight percent ranges depicted in Table D. It is understood that each and
every combination
of such components and weight percentages is intended the same as if each and
every
combination of component and weight percentage were specifically and
individually listed.
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Table D. Exemplary Weight Percent of Certain Components for Use in Extended
Release
Formulations
Formulation Exemplau Specific components Exemplary w/w %
Component
Therapeutic mannosamine, N-acetylmannosamine From about 20 to about 80;
Agent (MariNAc), ManNac-6-phosphate from about 20 to about 60;
(A compound (ManNAc-6-P), LIDP-GleNAc, N- from about 20 to about 50;
in the sialic acetylneuraminic acid (Neu.Ac), Netlike- from about 20 to
about 40;
acid 9-phosphate (NeuAc-9-P), sialic acid from about 20 to about
30;
biosynthetic 5-N-acetylneuram iTliC acid), CMP- from about 15 to about
60;
pathway or sialic acid, and/or derivatives thereof or from about 15 to
about 50;
derivative pharmaceutically acceptable salts of the from about 15 to
about 40;
thereof or salt foregoing. from about 25
to about 60;
of any of the from about 25 to about 50;
foregoing) from 25 to about 40; from
about 25 to about 30; from
about 30 to about 60; from
about 30 to about 50; from
about 30 to about 45; from
about 30 to about 40; from
about 35 to about 60; from
about 35 to about 50; from
about 35 to about 45; from
about 40 to about 45; about any
of 35, 36, 37, 38, 39, 40, 41,
42, 43, 44, 45, 46, 47, 48, 49
and 50.
Polymer I Carrageenan (e.g,, iota, kappa or lambda From about 1 to
about 10; from
(Hydrocolloid carrageenan, or a salt thereof, such as about 1 to about
5; from about
Polymer) Viscarin GP-209, FMC BioPolymer) 3 to about 8; from about 4
to
about 6; about any of 1, 2, 3, 4,
5, 6, 7, 8, 9 and 10.
Polymer 2 Alginate or salt thereof (e.g., sodium, From about 15 to
about 30;
(Anionic, pH- potassium or ammonium alginate salt from about 15 to about
25;
dependent , gel such as Protanalt, FMC BioPolymer), from about 15 to about
20;
forming co- carboxymethyl cellulose or salt thereof from about 20 to
about 30;
polymer) (e.g., sodium carboxymeth.y1 cellulose), from about 20 to
about 25;
from about 2o to about 23;
about any of 15, 16, 17, 18, 19,
20, 21, 22, 23, 24 and 25.
Polymer 3A Hyprom.ellose (e.g., hypromellose Type From about 20 to about
50;
(Water- 2208, E and K series such as for example, from about 20 to about
40,
swellable, pH Dow Chemical Company's (Midland, from about 20 to about
30;
independent Mich. USA) or Aqualon's (with a North from about 20 to about
25;
polymer) American presence in Wilmington, Del.) from about 25 to about
30;
E4M, E10M, K1OOLV, K4M, K.15M, from about 22 to about 27;
K25M, K100M, K200M and mixtures of about any of 20, 21, 22, 23, 24,
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Formulation Exemplary Specific components .Exemplary w/w %
component
various molecular weights and grades); 25, 26, 27, 28, 29 and 30.
hydroxypropyl ethyl cellulose (Ethocel
polymers 7FP, 10FP and 100FP and
Aqualon's polymersrflOEC, N7, N10,
N17, N22, N50, N100 and N200, and
mixtures thereof), hydroxypropyl
cellulose (Aqualon's HPC polymers MF
and MXF (mol. wt. 580,000) and KF and
HXF (mol. wt. 1,150,000), and mixtures
thereof); hydroxyethyl. cellulose (e.g.,
Aqualon's Natrasol polymers HHX (mol.
Wt. 1,300,000), FIX (mol. wt. 1,000,000),
H (mol. wt. 1,000,000), M (mol. wt.
720,000 and G (mol. wt. 1,150,000), and
mixtures thereof); methyl cellulose..
Polymer 3B Polyhydroxyethyle methylacrylate From about 20 to about 30;
(Hydrogel- (PHEMA), polyvinyl alcohol (PVA), from about 20 to about 25;
forming polyvinyl pyrrolidone (PVP), from about 25 to about 30;
polymer) polyethylene oxide (PEO), from about 22 to about 27;
polyacrylamide (PA), polyethylene oxide about any of 20, 21, 22, 23, 24,
(e.g., PolyoxTM water soluble resin, Dow 25, 26, 27, 28, 29 and 30.
Chemical Company, Mich., USA).
Excipient lactose, m.icrocrystall.in.e cellulose, corn From. about 1 to
about 20 or
starch, potato starch, wheat starch, about 1 to about 15 or about
1
sucrose, D-mannitol, precipitated calcium to about 10 or about 1 to about
carbonate, dextrin, pre-gelatinized starch, 5 or about 5 to about 20 or
microcrystalline cellulose and colloidal about 5 to about 15 or about
5
silicon dioxide (e.g., ProSolv SMCC to about 10 or about any one
of
HD90) and combinations thereof. 1, 2, 3, 4, 5, 6, 7, 8, 9 or
10
weight percent.
Lubricant Stearate salt (such as magnesium stearate From about 0.1 to about
2 or
(e.g., HyQuale) and calcium stearate), about 0.1 to about 1.5or
about
talc, light anhydrous sili.cic acid, and 0.1 to about 1.0 or about .01
to
solid polyethyl glycols and combinations about .09-5 or about 0.1 to
thereof. about 0.8 or about 0.1 to
about
0.7 or about 0.1 to about 0.6 or
about 0.1 to about 0.5 or about
0.2 to about 0.8 or about 0.3 to
about 0.7 or about 0.4 to about
0.6 or about any one of 0.1,
0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8,
0.9 or 1.0 weight percent.
[0011711 in another variation, extended release formulations are provided
wherein the
formulation comprises a therapeutic agent of Table C, a polymer 1 of Table C,
a polymer 2 of
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Table C, a Polymer 3A or a polymer 3B of Table C, an excipient of Table C and
a lubricant
of Table C, wherein the components are present in the composition in the
following weight
percent ratios. In
one aspect, the weight percent ratio of Lubricant:Poiymer
1:Excipient:Polymer 2:Polymer 3A or 3B:Therapeutic Agent is about
1:8:10:40:50:85 or
about 1:8.5:10,5:42.5:51:86.5 or about 1.:8.4:10.6:42.4:51:86.6.
100118] In
any of the formulae detailed herein, including but not limited to the
formulations in any of Tables A-D, in one aspect the therapeutic agent is
sialic acid or
ManNAc or a pharmaceutically acceptable salt thereof or a combination of
sialic acid or
ManNAc. In a particular aspect of any of the formulations detailed herein,
including but not
limited to the formulations in any of Tables A-D, the therapeutic agent is
sialic acid, or a
pharmaceutically acceptable salt thereof.
[00119]
Particular extended release formulations of sialic acid are provided in Table
E. In
one variation, ManNAc may be used in place of sialic acid in the formulations
of Table E.
Table E. Exemplary Extended Release Formulations of sialic acid.
Formulation Component Exemplary w/w %
Sialic acid, or From about 20 to about 80; from about 20 to about 60;
from
pharmaceutically about 20 to about 50; from about 20 to about 40; from
about 20 to
acceptable salt thereof about 30; from about 15 to about 60; from about 15
to about 50;
from about 15 to about 40; from about 25 to about 60; from about
25 to about 50; from 25 to about 40; from about 25 to about 30;
from about 30 to about 60; from about 30 to about 50; from about
30 to about 45; from about 30 to about 40; from about 35 to about
60; from about 35 to about 50; from about 35 to about 45; from
about 40 to about 45; about any of 35, 36, 37, 38, 39, 40, 41, 42,
43, 44, 45, 46, 47, 48, 49 and 50.
Carrageenan (e.g,, From about 1 to about 10; from about 1 to about 5; from
about 3
lambda carrageenan such to about 8; from about 4 to about 6; about any of 1,
2, 3, 4, 5, 6, 7,
as Viscarin GP-209, 8, 9 and 10.
MC BioPo ym er)
Alginate or a salt thereof From about 15 to about 30; from about 15 to about
25; from
(e.g., sodium, potassium about 15 to about 20; from about 20 to about 30; from
about 20 to
or ammonium alginate about 25; from about 2o to about 23; about any of 15,
16, 17, 18,
salt such as Protanal 19, 20, 21, 22, 23, 24 and 25.
LF 120M)
Hypromellose (such as From about 20 to about 50; from about 20 to about 40,
from
hypromellose Type about 20 to about 30; from about 20 to about 25; from
about 25 to
2208, e.g., about 30; from about 22 to about 27; about any of 20,
21, 22, 23,
Methoce101(100 M 24, 25, 26, 27, 28, 29 and 30.
Premium CR)
Polyethylene oxide (such From about 20 to about 30; from about 20 to about 25;
from
as Polyethylene Oxide about 25 to about 30; from about 22 to about 27;
about any of 20,
WSR, e.g., PolyoxTm 21, 22, 23, 24, 25, 26, 27, 28, 29 and 30.
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Formulation Component Exemplary w/w %
water soluble resin, Dow
Chemical Company,
M ic h., USA)
Microcrystalline From about 1 to about 20 or about 1 to about 15 or about
1 to
cellulose and colloidal about 10 or about 1 to about 5 or about 5 to about
20 or about 5 to
silicon dioxide (e.g., about 15 or about 5 to about 10 or about any one of
1, 2, 3, 4, 5,
FroSolv SMCC HD90) 6, 7, 8, 9 or 10 weight percent.
Stearate salt (such as From about 0.1 to about 2 or about 0.1 to about
1.5or about 0.1 to
magnesium stearate (e.g., about 1.0 or about .01 to about .09-5 or about 0.1
to about 0.8 or
HyQual0) about 0.1 to about 0.7 or about 0.1 to about 0.6 or
about 0.1 to
about 0.5 or about 0.2 to about 0.8 or about 0.3 to about 0.7 or
about 0.4 to about 0.6 or about any one of 0.1, 0.2, 0.3,0.4, 0.5,
0.6, 0.7, 0.8, 0.9 or 1.0 weight percent.
1001201 The components used to formulate the extended release pharmaceutical
formulations are preferably of high purity and are substantially free of
potentially harmful
contaminants (e.g., at least National Food (NW) grade, generally at least
analytical grade, and
more typically at least pharmaceutical grade). Moreover, pharmaceutical
formulations
intended for in vivo use are usually sterile. To the extent that a given
compound must be
synthesized prior to use, the resulting product is typically substantially
free of any potentially
toxic agents which may be present during the synthesis or purification
process. Compositions
for parental administration are also sterile, substantially isotonic and made
under GMP
conditions.
[00121] The blends of the extended release formulation may have a particle
size with a
majority of particles being retained by a sieve size of 45 p.m. In some
embodiments, the
blends of the extended release formulation have a particle size with at least
any one of 10%,
30%, 40%, 50% of particles retained by a sieve size of 45 prn.
[00122] The extended release pharmaceutical formulations as described herein
may be
formulated into preparations in solid, semi-solid, liquid or gaseous forms,
such as tablets,
capsules, powders, granules, ointments, solutions, suppositories, injections,
inhalants, gels,
microspheres, and aerosols. In some embodiments of any of the extended release
pharmaceutical formulations described herein, the extended release
pharmaceutical
formulations is formulated for administration by a variety of routes including
oral, parenteral
(including subcutaneous, intravenous, intramuscular and intraperitoneal),
rectal, dermal,
transdermal, intrathoracic, intrapulmonary and intranasal (respiratory)
routes. In some
embodiments, the extended release pharmaceutical formulation is formulated for
oral
administration.
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[00123] Any of the extended release formulations detailed herein may in one
variation be
formulated for oral administration. For example, any of the formulations
provided under the
heading "Extended Release Formulations," including but not limited to any of
the
formulations set forth in Tables A-E, Example 6, or Example 7 may in one
variation be a
formulation that is suitable for oral administration. A formulation that is
suitable for oral
administration may be formulated as a solid oral dosage form, such as a tablet
or a capsule
comprising the formulation as a powder. In one aspect, a solid oral dosage
form of an
extended release formulation is provided wherein the solid oral dosage form
comprises any
formulation provided herein (including but not limited to the formulations set
forth in any one
of Tables A-E, Example 6, or Example 7) in tablet form, wherein the tablet
further comprises
a coating (e.g., Opadiy-II White). In one variation, the extended release
formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof, as the
therapeutic agent.
In another variation, the extended release formulation comprises MaNAc, or a
pharmaceutically acceptable salt thereof, as the therapeutic agent. In another
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof and MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agents.
In another variation, the extended release formulation comprises a prodrug of
one or more
compounds in the sialic acid biosynthetic pathway such as a prodrug of sialic
acid, or a
pharmaceutically acceptable salt thereof, as the therapeutic agent. In another
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agent and further comprises a hydrocolloid
polymer, an anionic,
pH-dependent gel forming co-polymer and a water swellable, pH independent
polymer and
optionally further comprises a lubricant and/or an excipient. In a particular
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, carrageenan, sodium al.gin.ate and either hypromellose or
polyethylene oxide. in a
further variation, the extended release formulation comprises sialic acid, or
a
pharmaceutically acceptable salt thereof, carrageenan, sodium al.ginate,
either hypromellose
or polyethylene oxide, magnesium stearate and microcrystalline cellulose and
colloidal
silicon dioxide. In one aspect, the extended release formulation is a
formulation of Table E.
In yet another aspect, the extended release formulation is a formulation of
Table 8. In yet
another aspect, the extended release formulation is a formulation of Example
6. In yet
another aspect, the extended release formulation is a formulation of Example
7.
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[00124] For oral administration, the sialic acid biosynthetic pathway or
derivative thereof
as described herein can be administered in solid dosage forms, such as
capsules, tablets, and
powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions.
[00125] In some embodiments, the pharmaceutical formulations comprise an
enteric-
coating. Numerous types of acid-resistant enteric coatings are available.
Examples of the
acid-resistant coatings include cellulose acetate phthalate, polyvinyl acetate
phthalate,
shellac, an acrylic acid homopolymer or copolymer, a methaciylic acid
homopolymer or
copolymer, cellulose acetate trimellitate, hydroxypropyl methylcellulose
phthalate or a
combination of thereof. A number of copolymers of m.ethamylic acid are known
in the art and
are commercially available. Examples of such polymers are copolymers of
methylmethacrylate and methacrylic acid and copolymers of ethylacrylate and
methacrylic
acid, and sold under the tradename Eudragit (Rohm GmbH & Co. KG): examples
include
Eudragit L 100-55, Eudragit L 30D-55, Eudragit L 100, Eudragit S 100-55
and
Eudragit FS 30D. In some embodiments, the enteric coating comprises one or
more of
titanium dioxide, polydextrose, hypromellose, triacetin and macrogoUPEG. In
some
embodiments, the enteric coating is Opadry II White. In some embodiments, the
enteric
coating (e.g., Opadry II White) comprises about 1-5% w/w of the extended
release
formulation. In some embodiments, the enteric coating (e.g., Opadry II White)
comprises
about 1-5% w/w of the extended release formulation.
[00126] An enteric coating can also be a time-release coating. The time-
release coatings
are degraded away at a relatively constant rate until the coatings dissolve
sufficiently for the
time-release coatings to rupture. Thus, the ti.m.e required for the rupture of
the enteric coatings
is largely time-dependent (i.e., thickness), and largely pH independent.
Examples of time-
release coating materials include cellulose acetate, cellulose acetate
butyrate, cellulose acetate
propionate, EC, and copolymers of acrylate and m.ethacrylates with quaternary
ammonium
groups such as Eudragit RL and Eudragit RS and Eudragit NE30-D.
[00127] The extended release pharmaceutical formulations can be further
subjected to a
process of film coating. For the film coating agent, an enteric or non-enteric
film coating
agent may be used, and the enteric film, coating agent can be cellulose
acetate phthalate
(CAP), polyvinyl acetate phthalate (PVAP), a methacrylate polymer (Eudragit L,
S), or the
like, while the non-enteric film coating agent can be hydroxypropyl.cellul.ose
(HPC), MC, EC,
HPMC, povidone, PVA, CA, shellac, or the like. The process of coating can be
performed by,
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for example, a pan coating method, a fluidized bed coating method, a
compression coating
method, or the like.
[00128] Coated tablets of the extended release formulation may be prepared in
various
sizes. For example, the coated tablets may have a length of about 16-20 mm, a
width of about
7-12 mm and a thickness of about 5-8 mm. In some embodiments, the coated
tablets have a
length of about 17.7 mm, a width of about 9.1 mm and a thickness of about 6.7
mm. In some
embodiments, the coated tablets have a length of about 19.3 mm, a width of
about 9.7 mm
and a thickness of about 8.0 mm.
[00129] In embodiments of any of the methods, the extended release formulation
comprises a drug load of about 30-60% (e.g., sialic acid and/or ManNAc), about
20-30% w/w
of a water-swellable, pH independent polymer (e.g. hypromellose), about 20-25%
w/w of an
anionic, pH-dependent, gel-forming copolymer (e.g., an alginate salt), about 1-
5 A) wlw of a
hydrocolloid polymer (e.g., a carrageenan), about 1-10% w/w of
microcrystalline cellulose
and colloidal silicon dioxide (e.g., Prosolv SMCC HD90), about 0.1-1% w/w/
magnesium
stearate (e.g. HyQual ), and about 1-5% of an enteric coating (e.g. Opadry II
White). In
some embodiments, the extended release formulation comprises a drug load of
about 30-60%
(e.g., sialic acid and/or ManNAc), about 20-30% w/w hypromellose (e.g.
hypromellose Type
2208 or Methocel. KIOOM), about 20-25% w/w sodium alginate (e.g. Protanal),
about 1-5%
w/w lambda carrageenan (e.g. Viscarin GP-209), about 1-10% w/w of
microcrystalline
cellulose and colloidal silicon dioxide (e.g., Prosolv SMCC HD90), about 0.1 -
1% w/w/
magnesium stearate (e.g. HyQual0), and about 1 - 5% of an enteric coating
(e.g. Opadry II
White). In some embodiments, the extended release formulation comprises a drug
load of
about 30-60% (e.g., sialic acid and/or ManNAc), about 25% w/w of a water-
swellable, pH
independent polymer (e.g. hypromellose), about 21% w/w of an anionic, pH-
dependent, gel-
forming copolymer (e.g., an alginate salt), about 4% w/w of a hydrocolloid
polymer (e.g., a
carrageenan), about 5% w/w of microcrystalline cellulose and colloidal silicon
dioxide (e.g.,
Prosolv SMCC HD90), about 0.5% w/w/ magnesium stearate (e.g., HyQual0), and
about
3.5% of an enteric coating (e.g., Opadry II White). In some embodiments, the
extended
release formulation comprises a drug load of about 30-60% (e.g., sialic acid
and/or
ManNAc), about 25% wlw hypromellose (e.g., hypromellose Type 2208 or Methocel
1(100M), about 21% w/w sodium. alginate (e.g. Protanal), about 4% w/w lambda
carrageenan
(e.g. Viscarin GP-209), about 5% w/w of microcrystalline cellulose and
colloidal silicon
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WO 2013/109906 PCT/US2013/022167
dioxide (e.g, Pros lv SMCC HD90), about 0.5% w/w/ magnesium stearate (e.g.
HyQuale), and about 3.5% of an enteric coating (e.g. Opadry II White).
[00130] in embodiments of any of the methods, the extended release formulation
comprises a drug load of about 30-60% (e.g., sialic acid and/or ManNAc), about
20-30% w/w
of a hydrogel (e.g. polyethylene oxide), about 20-25% w/w of an anionic, pH-
dependent, gel-
forming copolymer (e.g., an alginate salt), about 1-5 % w/w of a hydroconoid
polymer (e.g.,
a carrageenan, about 1-10% w/w of microcrystalline cellulose and colloidal
silicon dioxide
(e.g., ProsoIve SMCC HD90), about 0.1-1% w/w/ magnesium stearate (e.g.
HyOu.a10), and
about 1-5% of an enteric coating (e.g. Opadry if White). In some embodiments,
the
extended release formulation comprises a drug load of about 30-60% (e.g.,
sialic acid and/or
ManNAc), about 20-30% w/w polyethylene oxide (e.g. Polyox WSR), about 20-25%
w/w
sodium alginate (e.g. Protanal), about 1-5% w/w lambda carrageenan (e.g.
Viscarin GP-209),
about 1-10% w/w of microcrystalline cellulose and colloidal silicon dioxide
(e.g., Prosolv
SMCC HD90), about 0,1 - 1% w/w/ magnesium stearate (e.g. HyQuale), and about 1
- 5% of
an enteric coating (e.g. Opadry II White). In some embodiments, the extended
release
formulation comprises a drug load of about 30-60% (e.g., sialic acid and/or
ManNAc), about
25% w/w of a hydrogel (e.g. polyethylene oxide), about 21% w/w of an anionic,
pH-
dependent, gel-forming copolymer (e.g., an alginate salt), about 4% w/w of a
hydrocolloid
polymer (e.g., a carrageenan), about 5% w/w of microcrystalline cellulose and
colloidal
silicon dioxide (e.g., Prosolve SMCC HD90), about 0.5% w/w/ magnesium stearate
(e.g.
HyQuale), and about 3.5% of an enteric coating (e.g. Opadry Ii White). In
some
embodiments, the extended release formulation comprises a drug load of about
30-60% (e.g.,
sialic acid and/or ManNAc), about 25% w/w polyethylene oxide (e.g. Polyos
WSR), about
21% w/w sodium alginate (e.g. Protanal), about 4% w/w lambda carrageenan (e.g.
Viscarin
GP-209), about 5% w/w of microcrystalline cellulose and colloidal silicon
dioxide (e.g.,
Pros lve SMCC HD90), about 0.5% w/w/ magnesium stearate (e.g. HyQuale), and
about
3.5% of an enteric coating (e.g. Opadry 11 White),
Embodiments of Formulations
[00131] in one embodiment, the extended release formulation comprises
about 25% to
about 50% wlw of a sialic acid, or a pharmaceutically acceptable salt,
solvate, or ester
thereof; about 20% to about 40% w/w of one or more water-swellable, pH
independent
polymers or one or more hydrogel-forming polymers; about 15% to about 30% wlw
of one or
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more anionic, pH-dependent, gel-forming polymers; and about 3% to about 8% w/w
of one or
more hydrocolloid polymers or one or more cationic polymers.
[00132] In one embodiment, the extended release formulation comprises
about 25% to
about 50% w/w of a sialic acid, or a pharmaceutically acceptable salt,
solvate, or ester
thereof; about 20% to about 30% w/w hypromellose; about 3% to about 8% w/w
carrageenan; and about 20% to about 25% w/w sodium alginate. In one
embodiment, the
extended release formulation comprises about 30% to about 45% w/w of a sialic
acid. In one
embodiment, the extended release formulation comprises about 22% to about 27%
w/w
hypromellose. In one embodiment, the extended release formulation comprises
about 4% to
about 6% w/w carrageenan. In one embodiment, the extended release formulation
comprises
about 20% to about 23% w/w carrageenan. In one embodiment, the extended
release
formulation further comprises about 1% to about 10% w/w of the mixture of
rnicrocrystalline
cellulose and colloidal silicon dioxide; and about 0.1% to about 1% w/w
magnesium stearate.
[00133] In one embodiment, the extended release formulation comprises
about 25% to
about 50% w/w of a sialic acid, or a pharmaceutically acceptable salt,
solvate, or ester
thereof; about 20% to about 30% w/w at least one of polyethylene glycol and
polyethylene
oxide; about 3% to about 8% w/w carrageenan; and about 20% to about 25% w/w
sodium
alginate. In one embodiment, the extended release formulation comprises about
30% to
about 45% w/w of a sialic acid. In one embodiment, the extended release
formulation
comprises about 22% to about 27% w/w at least one of polyethylene glycol and
polyethylene
oxide. In one embodiment, the extended release formulation comprises about 4%
to about
6% w/w carrageenan. In one embodiment, the extended release formulation
comprises about
20% to about 23% wlw carrageenan. In one embodiment, the extended release
formulation
further comprises about 1% to about 10% w/w of the mixture of microctystalline
cellulose
and colloidal silicon dioxide; and about 0.1% to about 1% w/w magnesium
stearate.
[00134] In one specific embodiment, the extended release formulation
comprises about
43.3% of a sialic acid, or a pharmaceutically acceptable salt, solvate, or
ester thereof; about
25.5% w/w hypromellose; about 4.2% w/w carrageenan; about 21.1% w/w sodium
alginate;
about 5.3% microcrystalline cellulose and colloidal silicon dioxide; and about
0.5%
magnesium stearate. In another embodiment, the extended release formulation is
in 325 mg
and 500 mg dosage form.
Therapeutic Utilities
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[00135] The present invention provides a method for treating a sialic acid
deficiency in an
individual in need thereof. The method comprises orally administering a sialic
acid, or a
pharmaceutically acceptable salt, solvate, or ester thereof, and provides a
therapeutically
effective amount of sialic acid over a period of greater than about four
hours.
[00136] in one embodiment, the sialic acid, or a pharmaceutically acceptable
salt, solvate,
or ester thereof, is in an extended release formulation, such as the ones
described herein. In
another embodiment, the sialic acid, or a pharmaceutically acceptable salt,
solvate, or ester
thereof, is in both an extended release formulation, such as the ones
described herein, and an
immediate release formulation. The extended release formulation and the
immediate release
formulation can be in separate dosage forms and be administered in a
coordinated fashion.
For example, an individual can take a dosage form of the extended release
formation and a
dosage form of the immediate release formulation concomitantly. Alternatively,
extended
release formulation and the immediate release formulation can be formulated in
a single
dosage form. For example, a single dosage form of a sialic acid, or a
pharmaceutically
acceptable salt, solvate, or ester thereof, can comprise an extended release
component and an
immediate relapse component.
[00137] In one embodiment of the present method, the sialic acid, or a
pharmaceutically
acceptable salt, solvate, or ester thereof, is administered on a regular
dosing schedule having
one or more dosing intervals per day. For example, the sialic acid, or a
pharmaceutically
acceptable salt, solvate, or ester thereof, can be administered once, twice,
three or four times
per day. In one embodiment, sialic acid, or a pharmaceutically acceptable
salt, solvate, or
ester thereof, is administered three times per day (TID).
[00138] in one embodiment, the present method provides a therapeutically
effective
amount of sialic acid over a period of greater than about eight hours per day.
In some
embodiments, the present method provides a therapeutically effective amount of
sialic acid
over a period of greater than about ten, twelve, fourteen, sixteen, or
eighteen hours per day.
For example, the present method provides a therapeutically effective amount of
sialic acid
over a period of about eight to about ten, about eight to about twelve, about
eight to about
fourteen, about eight to about sixteen, about ten to about fourteen, about
twelve to about
sixteen, or about sixteen to about twenty hours per day.
[00139] In one embodiment, the present method provides a mean Cmin sialic acid
of at least
about 0.11 meg/nil at steady state during the dosing intervals. In some
embodiments, the
present method provides a mean Cmin sialic acid of at least about 0.12 mcg/ml,
about 0.13
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mcg/rnl, about 0.14 mcg/ml, about 0.15 mcg/ml, about 0.16 mcg/ml, about 0.17
mcg/ml at
steady state during the dosing intervals.
[00140] In one embodiment, the present method provides a mean plasma
concentration of
sialic acid of at least about 0.16 mcg/m1 at steady state during the dosing
intervals. In some
embodiments, the present method provides a mean plasma concentration of sialic
acid of at
least about 0.17 mcg/ml, about 0.18 mcg/ml, about 0.19 mcg/ml, about 0.20
mcg/ml, about
0.21 mcg/m1., about 0.22 mcg/ml, about 0.23 mcg/ml, about 0.24 mcg/ml at
steady state
during the dosing intervals.
[00141.] In one embodiment, the present method provides a mean plasma
concentration of
sialic acid at steady state during the dosing intervals that is at least about
50% higher than the
mean plasm.a concentration of sialic acid in the individual before the
administration of the
sialic acid, or a pharmaceutically acceptable salt, solvate, or ester thereof.
In some
embodiments, the present method provides a mean plasma concentration of sialic
acid at
steady state during the dosing intervals that is at least about 60%, 70%, 80%,
90%, 100%,
110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, or 200% higher than the
mean
plasma concentration of sialic acid in the individual before the
administration of the sialic
acid, or a pharmaceutically acceptable salt, solvate, or ester thereof.
[00142] In one embodiment, the present method provides a plasma concentration
profile of
sialic acid at steady state such that the minimum plasma concentration of
sialic acid during
the dosing interval is at least about 35% of the maximum plasma concentration
during the
dosing interval. In some embodiments, the present method provides a plasma
concentration
profile of sialic acid at steady state such that the minimum, plasma
concentration of sialic acid
during the dosing interval is at least about 40%, 45%, 50%, 55%, or 60% of the
maximum
plasma concentration during the dosing interval.
[00143] In one embodiment, the present method provides an improved absorption
profile
when the extended release formulation is administered under fed conditions
than being
administered under fasting conditions. in one embodiment, the improved
absorption profile
includes that the mean C. determined at a fasted state is higher than the mean
Cu=
determined at a fed state. For example, the mean C. determined at a fasted
state is about
10%, 15%, 20%, 25%, 30%, or 35% higher than the mean C. determined at a fed
state. In
one embodiment, the improved absorption profile includes that the mean T.
determined at a
fed state is higher than the mean T. determined at a fasted state. For
example, the ratio of
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the mean Tina, determined at a fed state and the mean Tõx determined at a
fasted state is
about L2:1; 1.3:1; 1A:1; 15:1; 1.6:1; 1.7:1; 1.8:1; 1.9:1; or 2:1.
[00144] In one embodiment of the present invention, the extended release
formulations
detailed herein, including but not limited to those detailed under the heading
"Extended
Release Formulation" and "Additional Formulation Components" (e.g., any of the
formulations of Tables AE, Example 6 or Example 7) may exhibit any of the
characteristics
detailed herein and below, in a particular variation, any of the extended
release formulations
detailed herein may exhibit any one or more of the following characteristics:
(i) capable of
delivering to an individual in need thereof a therapeutically effective amount
of one or more
compounds in the sialic acid pathway or derivatives thereof over a period of
about or greater
than about any of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 hours;
(ii) capable of
delivering to an individual in need thereof a substantially constant
therapeutically effective
amount of one or more compounds in the sialic acid pathway or derivatives
thereof over a.
period of about or greater than. about any of 8, 9, 10, 11, 12, 13, 14, 15,
16, 17, 18, 19, or 20
hours; (iii) capable of delivering to an individual in need thereof a
therapeutically effective
amount of one or more compounds in the sialic acid pathway or derivatives
thereof with a
l',õõõ of between about any of 2-6 hours, 2-5 hours, or 3-6 hours during each
dosing interval;
(iv) capable of delivering to an individual in need -thereof a therapeutically
effective amount
of one or more compounds in the sialic acid pathway or derivatives thereof
with a C. of
about 0.1 ¨ 0.9 tg/m1,,, 0.1-100 [iglml, 0.2-0.3 Ring-, or 0.5-100 pg/mI.,;
(v) capable of
delivering to an individual in need thereof a therapeutically effective amount
of one or more
compounds in the sialle acid pathway or derivatives thereof with a trough
level of about 0.05
--- 0.2 lag/mL, 0.05 --- 0.3 [1g/rriL, 0.1 --- 0.3 lag/ML, or 0.1 20
_ig,/int. ; (vi) capable of
delivering to an individual in need thereof a therapeutically effective amount
of one or more
compounds in the sialic acid pathway or derivatives thereof with less than
about any of 10%,
20%, 30%, 40%, 50%, 60%, or 70% excreted after one hour; (vii) capable of
delivering to an
individual in need thereof between about any of 0.01-750 mg/kg/day, 0.5-500
mg/kg/day, 1-
250 mg/kg/day, 2.5-100 mg/kg/day, or 5-50 mg/kg/day of one or more compounds
in the
sialic acid pathway or derivatives thereof or a pharmaceutically acceptable
salt of the
foregoing; (viii) capable of delivering to an individual in need thereof
between about any of
0,01-750 mg/kg/day, 0.5-500 mg/kg/day, 1-250 mg/kg/day, 2.5-100 mg/kg/day, or
5-50
'mg/kg/day of one or more compounds in the sialic acid pathway or derivatives
thereof or a
pharmaceutically acceptable salt of the foregoing; (ix) has an absolute
bioavailabitity of about
CA 02862836 2014-07-02
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1 to about 5004); (x) has a bioavailability based on sialic acid levels in the
urine of about 0.5 to
about 100%; (xi) has a mean residence time (MRT) of at least about 3.5 hours.
In one
variation, the extended release formulation comprises sialic acid, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agent. In another variation, the
extended release
formulation comprises MaNAc, or a pharmaceutically acceptable salt thereof, as
the
therapeutic agent. In another variation, the extended release formulation
comprises sialic
acid, or a pharmaceutically acceptable salt thereof and MaNAc, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agents. In another variation, the
extended release
formulation comprises a prodrug of one or more compounds in the sialic acid
biosynthetic
pathway such as a prodrug of sialic acid, or a pharmaceutically acceptable
salt thereof, as the
therapeutic agent. In another variation, the extended release formulation
comprises sialic
acid, or a pharmaceutically acceptable salt thereof, as the therapeutic agent
and further
comprises a hydrocolloid polymer, an anionic, pH-dependent gel forming co-
polymer and a
water swellable, pH independent polymer and optionally further comprises a
lubricant and/or
an excipient. In a particular variation, the extended release formulation
comprises sialic acid,
or a pharmaceutically acceptable salt thereof, carrageenan, sodium alginate
and either
hypromellose or polyethylene oxide. In a further variation, the extended
release formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof,
carrageenan, sodium
alginate, either hypromellose or polyethylene oxide, magnesium stearate and
microcrystalline
cellulose and colloidal silicon dioxide. In one aspect, the extended release
formulation is a
formulation of Table E. In yet another aspect, the extended release
formulation is a
formulation of Table 8. In yet another aspect, the extended release
formulation is a
formulation of Example 6. In yet another aspect, the extended release
formulation is a
formulation of Example 7.
[00145.1 In embodiments of any of the extended release formulations, the
extended release
formulation is capable of delivering to an individual in need thereof a
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
over a period of greater than about any of 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, or 20
hours. In some embodiments, the extended release formulation is capable of
delivering to an
individual in need thereof a therapeutically effective amount of one or more
compounds in
the sialic acid pathway or derivatives thereof over a period of greater than
about 12 hours or
greater than about 24 hours. In embodiments of any of the extended release
formulations, the
extended release formulation is capable of delivering to an individual in need
thereof a
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therapeutically effective amount of one or more compounds in the sialic acid
pathway or
derivatives thereof over a period of between about any of 6-10 hours, 8-12
hours, 10-16, or
12-20 hours. In embodiments of any of the extended release formulations, the
extended
release formulation is capable of delivering to an individual in need thereof
a therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
over a period of about any of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or
20 hours. In some
embodiments, the extended release formulation is capable of delivering to an
individual in
need thereof a therapeutically effective amount of one or more compounds in
the sialic acid
pathway or derivatives thereof over a period of about 12 hours or about 24
hours. In some
embodiments, the therapeutically effective amount is delivered to the
bloodstream of the
individual. In some embodiments, the therapeutically effective amount is
delivered to muscle
tissue of the individual. In some embodiments, the one or more compounds in
the sialic acid
pathway or derivatives thereof include ManNAc or a derivative thereof and/or
sialic acid or a
derivative thereof. For example, in some embodiments, the extended release
formulation is
capable of delivering to an individual in need thereof a therapeutically
effective amount of
ManNAc and/or sialic acid to muscle tissue of the individual over a period of
between about
any of 6-10 hours, 8-12 hours, 10-16, or 12-20 hours. In one variation, the
extended release
formulation comprises sialic acid, or a pharmaceutically acceptable salt
thereof, as the
therapeutic agent. In another variation, the extended release formulation
comprises MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agent. In
another variation,
the extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof and MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agents.
[00146] In embodiments of any of the extended release formulations, the
extended release
formulation is capable of delivering to an individual in need thereof a
substantially constant
(i.e., without large burst of drug availability and deficiencies in drug
availability to the blood
and/or tissues of interest (e.g., muscle tissue)) therapeutically effective
amount of one or
more compounds in the sialic acid pathway or derivatives thereof over a period
of greater
than about any of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 hours.
In embodiments of
any of the extended release formulations, the extended release formulation is
capable of
delivering to an individual in need thereof a substantially constant
therapeutically effective
amount of one or more compounds in the sialic acid pathway or derivatives
thereof over a
period of between about any of 6-10 hours, 8-12 hours, or 10-16, or 12-20
hours. In
embodiments of any of the extended release formulations, the extended release
formulation is
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capable of delivering to an individual in need thereof a substantially
constant therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
over a period of about any of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or
20 hours. In some
embodiments, the one or more compounds in the sialic acid pathway or
derivatives thereof
include ManNAc or a derivative thereof and/or sialic acid or a derivative
thereof. For
example, in some embodiments, the extended release formulation is capable of
delivering to
an individual in need thereof a substantially constant therapeutically
effective amount of
ManNAc and/or sialic acid to muscle tissue of the individual over a period of
between about
any of 6-10 hours, 8-12 hours, or 10-16, 12-20 hours. In one variation, the
extended release
formulation comprises sialic acid, or a pharmaceutically acceptable salt
thereof, as the
therapeutic agent. In another variation, the extended release formulation
comprises MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agent. In
another variation,
the extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof and MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agents.
[00147] In embodiments of any of the extended release formulations, the
extended release
formulation is capable of delivering to an individual in need thereof a
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
with a C. of about 0.1-0.9 lag/mIõ 0.1-100 ti.g/m1õ 0.2-0.3 ttg/mL, or 0.5-100
ttg/mL. In
some embodiments, the extended release formulation is capable of delivering to
an individual
in need thereof a therapeutically effective amount of one or more compounds in
the sialic
acid pathway or derivatives thereof with a C. of about any one 0.5-80 1.1g/mL,
0.5-60
ttg/mL, 0.5 ¨ 40 1.1,g/m1, or 0.5 ¨ 20 ti.g/mL. In some embodiments, the
extended release
formulation is capable of delivering to an individual in need thereof a
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
with a C. of about 0.5 40 gg/mL. In some embodiments, the extended release
formulation
is capable of delivering to an individual in need thereof a therapeutically
effective amount of
one or more compounds in the sialic acid pathway or derivatives thereof with a
C. of about
any one of 0.5 ¨35 1.tg/mL, 0.5 ¨30 pg,/mL, 0.5 ¨25 p.g/mL, 1-40 ti.g/mL, 2.5
¨40 p.g/mL, 5
¨ 40 1.1,g/m.1õ 0.5 ¨ 35 pg/mL, 1 ¨ 35 p,g/mL, 2.5 ¨ 35 pg/m1õ 5 ¨ 35
pg/m1õ 0.5 ¨ 30 pg/ml.õ
1 ¨ 30 pg/mL, 2.5 ¨ 30 ilgimL, 5 ¨ 30 pg,/mL, 0.5 ¨ 25 pg,/mL, 0.1 ¨ 0.3
1.1g/mL, 0.1 ¨ 0.8
pg/mL, 0.2 ¨ 0.41ag/mIõ 0.2 ¨ 0.5 ti.g/mL, 0.2 ¨ 0.8 pg/mL, 0.1 ¨ liag/mIõ 1 ¨
25 tiglml.õ 2.5
¨ 25 ggirnL, or 5 ¨ 25 pg/mL. In some embodiments, the extended release
formulation is
capable of delivering to an individual in need thereof a therapeutically
effective amount of
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one or more compounds in the sialic acid pathway or derivatives thereof with a
C. of about
0.5 - 20 ttg/mL. In some embodiments, the extended release formulation is
capable of
delivering to an individual in need thereof a therapeutically effective amount
of one or more
compounds in the sialic acid pathway or derivatives thereof with a C. of about
0.1 - 1
1.1.g/mL. In some embodiments, the extended release formulation is capable of
delivering to
an individual in need thereof a therapeutically effective amount of one or
more compounds in
the sialic acid pathway or derivatives thereof with a C. of about any one of
0.5 - 15 1.1.g/m1L,
0.5 - 10 gg/mL, 1 - 20 itglinL, 2.5 - 20 pg/mL, 5 - 20 itg/mL, 0.5 - 15
ilg/mL, 1-15
pg/mL, 2.5 - 15 pg/mL, 5 - 15 Itg/mL, 0.5 - 10 ttglmL, 1 - 10 ug/mIõ 2.5 - 10
ug/mIõ or 5 -
ug/mL. In some embodiments, the one or more compounds in the sialic acid
pathway or
derivatives thereof include ManNAc and/or sialic acid. In some embodiments,
the one or
more compounds in the sialic acid pathway or derivatives thereof include
sialic acid. In one
variation, the extended release formulation comprises sialic acid, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agent. in another variation, the
extended release
formulation comprises MaNAc, or a pharmaceutically acceptable salt thereof, as
the
therapeutic agent. in another variation, the extended release formulation
comprises sialic
acid, or a pharmaceutically acceptable salt thereof and MaNAc, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agents.
[00148] In embodiments of any of the extended release formulations, the
extended release
formulation is capable of delivering to an individual in need thereof a
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
with a trough level of about 0.05 - 0.21ag/mIõ 0.05 -0.3 Itg/mL, 0.1 -0.3
pg/mIõ or 0.1 -20
1.1g/mL. In embodiments of any of the extended release formulations, the
extended release
formulation is capable of delivering to an individual in need thereof a
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
with a trough level of about any one of 0.05 - 0.2 1.tg/mL, 0.05 - 0.3 pg/m1õ
0.1 - 0.2 pg/mL,
0.1 -0.3 pg/mL, 0.2 0.3 p.g/mL, 0.1 15 ttg/mL, 0.1 10 ttg/mL, 0.1 5 ttg/m1,,
0.5 - 20,
1.1g/mL, 0.5 - 15 ggirnL, 0.5 - 10 g/mL, 0.5 5 gglmL, I 20 pg/mL, 1 - 15
1.1g/mL, 1 - 10
gg/m1õ or 1 - 5 pg/MI, or about any one of 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5,
4, 4.5, 5, 5.5, 6, 7, 8,
9, 10, 11, 12, 13, 14 or 15 ttg/rnL. In embodiments of any of the extended
release
formulations, the extended release formulation is capable of delivering to an
individual in
need thereof a therapeutically effective amount of one or more compounds in
the sialic acid
pathway or derivatives thereof with a trough level of about 0.05-0.3 ttglmL.
In some
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embodiments, the one or more compounds in the sialic acid pathway or
derivatives thereof
include ManNAc and/or sialic acid. In some embodiments, the one or more
compounds in the
sialic acid pathway or derivatives thereof include sialic acid. In one
variation, the extended
release formulation comprises sialic acid, or a pharmaceutically acceptable
salt thereof, as the
therapeutic agent. In another variation, the extended release formulation
comprises MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agent. In
another variation,
the extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof and MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agents.
[00149] In one embodiment of the present invention, the extended release
formulation is
administered three times a day (TID) to provide a therapeutic effect for
throughout the day,
i.e., about 24 hours. For example, the extended release formulation can be
taken by a patient
approximately every 8 hours on a daily basis. In one embodiment, the extended
release
formulation provides a mean maximum plasma concentration of sialic acid from
about 0.1 to
about 1 pg/mL from a mean of about 0.5 to about 6 hours after a first
administration. In
another embodiment, the extended release formulation provides a mean maximum
plasma
concentration of sialic acid from about 0.15 to about 0.85 pg/mL from a mean
of about 1 to
about 5.5 hours after a first administration. In another embodiment, the
extended release
formulation provides a mean maximum plasma concentration of sialic acid from
about 0.2 to
about 0.7 1.1.g/mL from a mean of about 1.5 to about 5 hours after a first
administration. In
another embodiment, the extended release formulation provides a mean maximum.
plasma
concentration of sialic acid from about 0.25 to about 0.55 pg/mL from a mean
of about 2 to
about 4.5 hours after a first administration. In another embodiment, the
extended release
formulation provides mean a maximum plasma concentration of sialic acid from
about 0.3 to
about 0.5 jAg/mL from a mean of about 2.5 to about 4 hours after a first
administration. In
one embodiment, the extended release formulation provides a mean minimum
plasma
concentration of sialic acid from about 0.1 to about 0.5 pg/rnL from a mean of
about 6 to
about 8 hours after repeated administration approximately every 8 hours
through steady-state
conditions. In another embodiment of the present invention, the extended
release formulation
provides a mean minimum plasma concentration of sialic acid from about 0.15 to
about 0.45
pg/mL from a mean of about 6 to about 8 hours after repeated administration
approximately
every 8 hours through steady-state conditions. In one embodiment of the
present invention,
the extended release formulation provides a mean minimum plasma concentration
of sialic
acid from about 0.2 to about 0.4 jAg/mL from a mean of about 6 to about 8
hours after
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repeated administration approximately every 8 hours through steady-state
conditions. In one
embodiment of the present invention, the extended release formulation provides
a mean
minimum plasma concentration of sialic acid from about 0.25 to about 0.35
1,tg,imL from a
mean of about 6 to about 8 hours after repeated administration approximately
every 8 hours
through steady-state conditions.
[00150] In embodiments of the present invention, the extended release
formulation, when
administered to a patient on a regular dosing schedule, provides to the
patient a therapeutic
effect continuously over the period of the regular dosing schedule. That is,
the therapeutic
effect, once attained after the first administration, is constant during the
period of the regular
dosing schedule which includes multiple dosing intervals. The regular schedule
can be a
dosing regimen provided by instructions accompanying the extended release
formulation
product. For example, such dosing regimen can be fixed or variable amount of
the extended
release formulation taken once per day, twice per day, three times per day, or
four times per
day.
[00151] In one embodiment, a total amount of about 650 mg to about 6000 mg
sialic acid,
or a pharmaceutically acceptable salt, solvate, or ester thereof, is
administered to a patient per
day on a regular dosing schedule. In another embodiment, a total amount of
about 1950 mg
to about 6000 mg sialic acid, or a pharmaceutically acceptable salt, solvate,
or ester thereof,
is administered to a patient per day on a regular dosing schedule. In one
embodiment, the
regular dosing schedule refers to one or more administrations with
approximately equal
dosing intervals in each dosing cycle. For example, a three times per day
(TID) dosing
schedule denotes administration of the drug three times per day with
approximately eight
hours dosing intervals. In one embodiment, about 650 mg of sialic acid, or a
pharmaceutically acceptable salt, solvate, or ester thereof, is administered
to a patient once
per day. In one embodiment, about 650 mg of sialic acid, or a pharmaceutically
acceptable
salt, solvate, or ester thereof, is administered to a patient on a three times
per day dosing
schedule. In one embodiment, about 650 mg of sialic acid, or a
pharmaceutically acceptable
salt, solvate, or ester thereof, is administered to a patient on a three times
per day dosing
schedule. In one embodiment, about 650 mg of sialic acid, or a
pharmaceutically acceptable
salt, solvate, or ester thereof, is administered to a patient on a three times
per day dosing
schedule (total amount of about 1950 mg per day). In one embodiment, about 975
mg of
sialic acid, or a pharmaceutically acceptable salt, solvate, or ester thereof,
is administered to a
patient on a three times per day dosing schedule (total amount of about 2925
mg per day). In
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one embodiment, about 1000 mg of sialic acid, or a pharmaceutically acceptable
salt, solvate,
or ester thereof, is administered to a patient on a three times per day dosing
schedule (total
amount of about 3000 mg per day). In one embodiment, about 1500 mg of sialic
acid, or a
pharmaceutically acceptable salt, solvate, or ester thereof, is administered
to a patient on a.
three times per day dosing schedule (total amount of about 4500 mg per day).
In one
embodiment, about 162.5 mg of sialic acid, or a pharmaceutically acceptable
salt, solvate, or
ester thereof, is administered to a patient on a three times per day dosing
schedule (total
amount of about 4875 mg per day). In one embodiment, about 2000 mg of sialic
acid, or a.
pharmaceutically acceptable salt, solvate, or ester thereof, is administered
to a patient on a
three times per day dosing schedule (total amount of about 6000 mg per day).
In one
embodiment, about 4000 mg of sialic acid, or a pharmaceutically acceptable
salt, solvate, or
ester thereof, is administered to a patient on a three times per day dosing
schedule (total
amount of about 12000 mg per day). In one embodiment, about 2000 mg of sialic
acid, or a.
pharmaceutically acceptable salt, solvate, or ester thereof, in extended
release fortnulaion and
about 2000 mg of sialic acid, or a pharmaceutically acceptable salt, solvate,
or ester thereof,
in immediate release formulaion are administered to a patient in combination
on a three times
per day dosing schedule (total amount of about 12000 mg per day).
[00152] In embodiments of the present invention, the extended release
formulation, when
administered to a patient on a regular dosing schedule, provides a relatively
flat plasma
concentration profile of sialic acid at steady state, wherein there are no
substantial peak or
trough in the relatively flat plasma concentration profile and the minimum
plasma
concentration of sialic acid in the relatively flat plasma concentration
profile is sufficient to
provide a therapeutic effect to the patient.
100153] In one embodiment of the present invention, the extended release
formulation,
when administered to a patient on a regular dosing schedule, provides a
relatively fiat plasma
concentration profile of sialic acid at steady state such that a mean
Cinin/C,õ,õ sialic acid ratio
during the dosing interval is about 0.40 to about LO, about 0.45 to about 1.0,
about 0,5 to
about 1.0, about 0.55 to about 1.0, about 0.6 to about 1.0, or about 0.65 to
about 1.0, or about
0.7 to about 1.0, or about 0.75 to about 1.0, or about 0,8 to about 1.0, or
about 0.85 to about
1.0, or about 0.9 to about 1.0, or about 0.95 to about 1.0 and the Cillin is
sufficient to provide a
therapeutic effect.
1001541 in one embodiment of the present invention, the extended release
formulation,
when administered to a patient a three times a day (T1D), provides a mean
C8/Cmax sialic acid
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ratio of about 0.40 to about 1.0, about 0.45 to about 1.0, about 0.5 to about
1.0, about 0.55 to
about 1.0, about 0.6 to about 1.0, or about 0.65 to about 1.0, or about 0.7 to
about 1.0, or
about 0.75 to about 1.0, or about 0.8 to about 1.0, or about 0.85 to about
1.0, or about 0.9 to
about 1.0, or about 0.95 to about 1.0 at steady state after administration to
the patient.
[00155] in one embodiment of the present invention, the extended release
formulation,
which is in a three times a day (TID) dosage form, provides to a patient a
therapeutic effect
for about 8 hours and a relatively flat plasma concentration profile of sialic
acid at steady
state such that the minimum plasma concentration of sialic acid during the
dosing interval is
about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the
maximum plasma concentration during the dosing interval.
[00156] In one embodiment of the present invention, the extended release
formulation,
which is in a three times a day (TID) dosage form, provides to a patient a
therapeutic effect
for about 8 hours and a relatively flat plasma concentration profile of sialic
acid at steady
state such that the maximum plasma concentration of sialic acid during the
dosing interval is
about 155%, 150%, 145%, 140%, 135%, 130%, 125%, 120%, 115%, 110%, or 105% of
the
minimum plasma concentration during the dosing interval.
[00157] In embodiments of any of the extended release formulations, the
extended release
formulation is capable of delivering to an individual in need thereof a
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
with less than about any of 10%, 20%, 30%, 40%, 50%, 60%, or 70% excreted
after one
hour. In embodiments of any of the extended release formulations, the extended
release
formulation is capable of delivering to an individual in need thereof a
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
with less than about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%
excreted
after four hours. in embodiments of any of the extended release formulations,
the extended
release formulation is capable of delivering to an individual in need thereof
a therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
with less than about any one of 2, 3, 4, or 5% excreted after 12 hours. In
some embodiments,
the one or more compounds in the sialic acid pathway or derivatives thereof
include ManNAc
or a derivative thereof and/or sialic acid or a derivative thereof. In some
embodiments, the
one or more compounds in the sialic acid pathway or derivatives thereof
include ManNA.c
and/or sialic acid. In some embodiments, the one or more compounds in the
sialic acid
pathway or derivatives thereof include sialic acid. In one variation, the
extended release
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formulation comprises sialic acid, or a pharmaceutically acceptable salt
thereof, as the
therapeutic agent. In another variation, the extended release formulation
comprises MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agent. In
another variation,
the extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof and MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agents.
[00158] In embodiments of any of the extended release formulations, the
extended release
formulation is capable of delivering to an individual in need thereof between
about any of
0.1-50 g/day, 0.5-25 g/day, 1-15 g/day, 1-10 g/day, or 2-5 g/day of one or
more compounds
in the sialic acid pathway or derivatives thereof. In some embodiments, the
extended release
formulation is capable of delivering to an individual in need thereof between
about 2 g/day
and 5 g/day of one or more compounds in the sialic acid pathway or derivatives
thereof. In
embodiments of any of the extended release formulations, the extended release
formulation is
capable of delivering to an individual in need thereof between about any of
0.01-750 mg/kg,
0.5-500 mg/kg, 1-250 mg/kg, 2.5-100 mg/kg, or 5-50 mg/kg of one or more
compounds in
the sialic acid pathway or derivatives thereof. In some embodiments, the
extended release
formulation is capable of delivering to an individual in need thereof between
about 5 mg/kg
and 50 mg/kg of one or more compounds in the sialic acid pathway or
derivatives thereof. In
some embodiments, the one or more compounds in the sialic acid pathway or
derivatives
thereof include ManNAc or a derivative thereof and/or sialic acid or a
derivative thereof. For
example, in some embodiments, the extended release formulation is capable of
delivering to
an individual in need thereof between about 5 mg/kg and 50 mg/kg of ManNAc
and/or sialic
acid. In one variation, the extended release formulation comprises sialic
acid, or a
pharmaceutically acceptable salt thereof, as the therapeutic agent. In another
variation, the
extended release formulation comprises MaNAc, or a pharmaceutically acceptable
salt
thereof, as the therapeutic agent. In another variation, the extended release
formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof and
MaNAc, or a
pharmaceutically acceptable salt thereof, as the therapeutic agents.
1001591 In embodiments of any of the extended release formulations, the
extended release
formulation is capable of delivering to an individual in need thereof between
about any of
0.01-750 mg/kg/day, 0.5-500 mg/kg/day, 1-250 mg/kg/day, 2.5-100 mg/kg/day, or
5-50
mg/kg/day of one or more compounds in the sialic acid pathway or derivatives
thereof. In
some embodiments, the extended release formulation is capable of delivering to
an individual
in need thereof between about 5 mg/kg/day and 50 mg/kg/day of one or more
compounds in
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the sialic acid pathway or derivatives thereof. In some embodiments, the one
or more
compounds in the sialic acid pathway or derivatives thereof include ManNAc or
a derivative
thereof and/or sialic acid or a derivative thereof. For example, in some
embodiments, the
extended release formulation is capable of delivering to an individual in need
thereof between
about 5 mg/kg/day and 50 mg/kg/day of ManNAc and/or sialic acid. In one
variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agent. In another variation, the extended release
formulation
comprises MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agent. In
another variation, the extended release formulation comprises sialic acid, or
a
pharmaceutically acceptable salt thereof and MaNAc, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agents.
1001601 In embodiments of any of the extended release formulations, the
extended release
formulation has an absolute bioavailability of about 1-50%. In some
embodiments, the
extended release formulation has an absolute bioavailability of about any one
of 1-45%, 1-
40%, 1-35%, 1-30%, 1-20%, 1-10%. In some embodiments the extended release
formulation
has an absolute bioavailability of about 1 25%. In some embodiments, the
extended release
formulation has an absolute bioavailability of about any one of 5, 10, 15, 20,
25 or 50%. In
some embodiments, the one or more compounds in the sialic acid pathway or
derivatives
thereof include ManNAc or a derivative thereof and/or sialic acid or a
derivative thereof. In
some embodiments, the one or more compounds in the sialic acid pathway or
derivatives
thereof include sialic acid or a derivative thereof. In one variation, the
extended release
formulation comprises sialic acid, or a pharmaceutically acceptable salt
thereof, as the
therapeutic agent. In another variation, the extended release formulation
comprises MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agent. In
another variation,
the extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof and MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agents.
[00161] in embodiments of any of the extended release formulations, the
extended release
formulation has a bioavailability based on sialic acid levels in the urine of
about 0.5-100%. In
some embodiments, the extended release formulation has a bioavailability based
on sialic
acid levels in the urine of about any one of 0.5-2.5%, 1-2.5 %, 2 - 8%, 2 -
12%, 2.5-20%,
2.5-40%, 2.5- 80%, 2.5 - 100%. In one variation, the extended release
formulation comprises
sialic acid, or a pharmaceutically acceptable salt thereof, as the therapeutic
agent. In another
variation, the extended release formulation comprises MaNAc, or a
pharmaceutically
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acceptable salt thereof, as the therapeutic agent. In another variation, the
extended release
formulation comprises sialic acid, or a pharmaceutically acceptable salt
thereof and MaNAc,
or a pharmaceutically acceptable salt thereof, as the therapeutic agents.
[00162] In embodiments of any of the extended release formulations, the
extended release
formulation has a mean residence time (MRT) of at least about 3.5 hours. In
som.e
embodiments, the extended release formulation has a MRT of at least about any
one of 3, 4,
4.5, 5, 5.5 or 6 hours. In some embodiments, the one or more compounds in the
sialic acid
pathway or derivatives thereof include ManNAc or a derivative thereof and/or
sialic acid or a
derivative thereof. In some embodiments, the one or more compounds in the
sialic acid
pathway or derivatives thereof include sialic acid or a derivative thereof. In
one variation, the
extended release formulation comprises sialic acid, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agent. In another variation, the extended release
formulation
comprises MaNAc, or a pharmaceutically acceptable salt thereof, as the
therapeutic agent. In
another variation, the extended release formulation comprises sialic acid, or
a
pharmaceutically acceptable salt thereof and MaNAc, or a pharmaceutically
acceptable salt
thereof, as the therapeutic agents.
[00163] The extended release formulations and/or the combination of the
extended release
formulation and immediate release formulation as described herein can be a
sialic acid
Reference Drug. The term "Reference Drug" is defined in the U.S. Federal Food
and Drug
Administration's (FDA) Orange Book, Approved Drug Products with Therapeutic
Equivalence Evaluations. The present invention includes any bioequivalence of
the extended
release formulations and/or the combination of the extended release
formulation and
immediate release formulation as a reference drug.
[00164] "Bioequivalence" denotes the absence of a significant difference in
the rate and
extent to which the active agent or surrogate marker for the active agent in
pharmaceutical
equivalents or pharmaceutical alternatives becomes available at the site of
action when
administered in an appropriately designed study. in one embodiment,
bioequivalence is any
definition thereof as promulgated by the U.S. Food and Drug Administration or
any successor
agency thereof. In a specific embodiment, bioequivalence is determined
according to the
Federal Drug Administration's guidelines and criteria, including "GUIDANCE FOR
INDUSTRY BIO.AV.AILABILIT.'Y AND BIOEQUVALENCE STUDIES FOR ORALLY
ADMINISTERED DRUG PRODUCTS¨GENERAL CONSIDERATIONS" available from
the U.S. Department of Health and Human Services (DHHS), Food and Drug
Administration
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(FDA), Center for Drug Evaluation and Research (CDER) March 2003 Revision 1;
and
"GUIDANCE FOR INDUSTRY STATISTICAL APPROACHES TO ESTABLISHING
BIOEQUIVAI.ENCE" DHHS, FDA, CDER, January 2001, both of which are incorporated
herein in their entirety. In another embodiment, bioequivalenee is determined
according to
the European Medicines Agency (EM EA) document "-Note for Guidance on the
Investigation
of Bioavailability and Bioequivalence", issued July 26, 2001, available from
EMEA.
1001651 In one embodiment, the present invention provides a formulation of
sialic acid, or
a pharmaceutically acceptable salt, solvate, or ester thereof; wherein the
formulation exhibits
a ratio of a geometric mean of logarithmic transformed AUC0.4 of the
fbrmulation to a
geometric mean of logarithmic transformed AUCo_t of the sialic acid Reference
Drug from
about 0.80 to about 1.25.
[00166] in one embodiment, the present invention provides a formulation of
sialic acid, or
a pharmaceutically acceptable salt, solvate, or ester thereof; wherein the
formulation exhibits
a ratio of a geometric mean of logarithmic transformed AUC0_õ. of the
formulation to a
geometric mean of logarithmic transformed AUC0_,, of the sialic acid Reference
Drug from
about 0.80 to about 1.25.
[00167] In one embodiment, the present invention provides a formulation of
sialic acid, or
a pharmaceutically acceptable salt, solvate, or ester thereof; wherein the
fbrmulation exhibits
a ratio of a geometric mean of logarithmic transformed Cmax of the formulation
to a geometric
mean of logarithmic transformed Cn. of the sialic acid R.etrence Drug from
about 0.80 to
about 1.25.
[00168] The extended release pharmaceutical formulation may be formulated for
parenteral administration (e.g., by injection, for example, bolus injection or
continuous
infusion) and may be presented in unit dose form in ampoules, pre-filled
syringes, small
volume infusion containers or in multi-dose containers. The extended release
pharmaceutical
formulation may be may form suspensions, solutions, or emulsions in oily or
aqueous
vehicles. Alternatively, the one or more compounds in the sialic acid
biosynthetic pathway or
derivative thereof and other ingredients may be in powder form, obtained by
aseptic isolation
of sterile solid or by lyophilization from solution, for constitution with a
suitable vehicle, e.g.,
sterile, pyrogen-free water, before use.
[00169] For topical administration, the extended release pharmaceutical
formulation may
be formulated as is known in the art for direct application to a target area.
Forms chiefly
conditioned for topical application take the form, for example, of creams,
milks, gels,
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dispersion or microemulsions, lotions thickened to a greater or lesser extent,
impregnated
pads, ointments or sticks, aerosol formulations (e.g., sprays or foams),
soaps, detergents,
lotions or cakes of soap. Other conventional forms for this purpose include
wound dressings,
coated bandages or other polymer coverings, ointments, creams, lotions,
pastes, jellies,
sprays, and aerosols. The extended release pharmaceutical formulation may be
delivered via
patches or bandages for dermal administration. Ointments and creams may, for
example, be
formulated with an aqueous or oily base with the addition of suitable
thickening and/or
gelling agents. Drops, such as eye drops or nose drops, may be formulated with
the one or
more compounds in the si.alic acid biosynthetic pathway or derivative thereof
in an aqueous
or non-aqueous base. Liquid sprays are conveniently delivered from pressurized
packs. Drops
can be delivered via a simple eye dropper-capped bottle, or via a plastic
bottle adapted to
deliver liquid contents dropwise, via a specially shaped closure.
[00170] Further, in some embodiments, the extended release pharmaceutical
formulation
comprising one or more compounds in the sial.ic acid biosynthetic pathway or
derivative
thereof may also be used in combination with other therapeutic agents.
Methods of Making Extended Release Formulations
[00171] Methods of making extended release formulations detailed herein are
also
provided. In one aspect, the formulation components (which may optionally be
delumped and
sieved to a desired range of particle size) are combined and mixed to provide
a uniform
formulation blend, which may further be used to prepare particular dosage
forms, such as
tablets or capsules, e.g., for oral administration. Particular dosage forms,
once prepared, may
be further modified to provide the final drug product, such as, e.g., by
administering a coating
to a tablet formed from an extended release formulation blend. Preparation of
the extended
release formulations may be accomplished through known techniques, such as
direct
compression, dry granulation and wet granulation.
[00172] Direct compression may be accomplished by delumping the formulation
components and sieving to a desired range of particle size, which may be the
same or
different size for individual formulation components. The components are then
blended,
which may be accomplished by one or a series of blending steps until all
formulation
components are blended. The blended formulation may, if desired, be direct
compressed to
provide the desired product, which may be in the form of a dosage suitable for
oral
administration, such as a tablet. The blended formulation may also be filled
into capsules or
other forms for solid-dosage administration, e.g., for oral administration.
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[00173] Dry granulation may also be utilized to prepare the extended release
formulations
detailed herein, and may be used to improve the flow or other characteristic
of a blend of
formulation components to be formed into a final drug product. One example of
dry
granulation includes delumping and/or sieving the formulation components,
blending the
formulation components and feeding the blend through, e.g., a roller compactor
that produces
a ribbon of compressed product, then milling the resulting ribbon. The milled
product may
then be compressed as detailed above or further blended with additional
formulation
components and compressed.
[00174] Wet granulation may also be utilized to prepare the extended release
formulations.
For example, the formulation components may be delumped and sieved to the
desired size,
and blended. The resulting blend may be added to an appropriate fluid bed
processor
equipped with a spray gun for fluidizing the blended formulation components
using standard
practices. The resulting granulation is dried (e.g., in the fluid bed) and
milled to a desired
range of particle sized and may be used for preparation of a final
formulation. Wet
granulation may also utilize high shear wet granulation (blended components
are mixed, and
frequently chopped while the solvent, typically water or other aqueous-based
solvent, is
sprayed over the mass during granulation).
[00175] Extended release formulations that are in tablet form preferably are
compressed to
a sufficient hardness to prevent premature ingress of a medium (e.g., aqueous
medium) and to
prevent surface pitting and breakage during coating, when applicable.
[00176] It is understood that extended release formulation blends are
provided, such as a
final formulation blend comprising a therapeutic agent and all formulation
components in a
final product (e.g., a blend comprising a therapeutic agent, a polymer, an
excipient and a
lubricant) as well as intermediate formulation blends that contain a portion
of all formulation
components in a final product (e.g., a blend comprising a therapeutic agent
and a polymer but
not an excipient or a lubricant, where the final product contains an excipient
and a lubricant).
Methods of Treating and Preventing Sialic Acid Deficiencies
[00177] Provided herein are also methods of treating and/or preventing sialic
acid
deficiencies in an individual in need thereof by administering an effective
amount of one or
more compounds in the sialic acid pathway or derivatives thereof in any
extended release
formulation described herein. The methods may comprise administration of an
effective
amount of any of the formulations detailed herein, including any of the
formulations under
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the heading "Extended Release Formulations," including but not limited to any
of the
formulations of Tables A-E as well as those formulations of Examples 12 and
13. Thus,
although certain formulations are detailed below, it is understood that any
extended release
formulations described herein may be employed in any of the methods provided
herein. In
one variation, the extended release formulation comprises sialic acid, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agent. In another variation, the
extended release
formulation comprises MaNAc, or a pharmaceutically acceptable salt thereof, as
the
therapeutic agent. In another variation, the extended release formulation
comprises sialic
acid, or a pharmaceutically acceptable salt thereof and MaNAc, or a
pharmaceutically
acceptable salt thereof, as the therapeutic agents. In another variation, the
extended release
formulation comprises a prodrug of one or more compounds in the sialic acid
biosynthetic
pathway such as a prodrug of sialic acid, or a pharmaceutically acceptable
salt thereof, as the
therapeutic agent. In another variation, the extended release formulation
comprises sialic
acid, or a pharmaceutically acceptable salt thereof, as the therapeutic agent
and further
comprises a hydrocolloid polymer, an anionic, pH-dependent gel forming co-
polymer and a
water swellable, pH independent polymer and optionally further comprises a
lubricant and/or
an excipient. In a particular variation, the extended release formulation
comprises sialic acid,
or a pharmaceutically acceptable salt thereof, carrageenan, sodium alginate
and either
hypromellose or polyethylene oxide. In a further variation, the extended
release formulation
comprises sialic acid, or a pharmaceutically acceptable salt thereof,
carrageenan, sodium
alginate, either hypromellose or polyethylene oxide, magnesium stearate and
microcrystalline
cellulose and colloidal silicon dioxide. In one aspect, the extended release
formulation is a
formulation of Table E. In yet another aspect, the extended release
formulation is a
formulation of Table 8. In another aspect, the extended release formulation is
a formulation
of Example 6. In another aspect, the extended release formulation is a
formulation of
Example 7. In some embodiments, the one or more compounds in the sialic acid
pathway or
derivatives thereof include ManNAc or a derivative thereof and/or sialic acid
or a derivative
thereof. For example, provided herein are methods of treating and/or
preventing sialic acid
deficiencies in an individual in need thereof by administering an effective
amount of
ManNAc and sialic acid in any extended release formulation described herein.
In some
embodiments, the methods of treating and/or preventing sialic acid
deficiencies increase
sialic acid production. In some embodiments, the methods of treating and/or
preventing sialic
acid deficiencies increase sialylation of affected tissue. In some
embodiments, the method of
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treating and/or preventing sialic acid deficiencies comprises administering an
extended
release formulation comprising a drug load of about 30-60% (e.g., sialic acid
and/or
ManNAc), about 20-30% w/w hypromellose (e.g. hypromellose Type 2208 or
Methocel
K100M), about 20-25% w/w sodium alginate (e.g. Protanal), about 1-5% w/w
lambda
carrageen.an (e.g. Viscarin GP-209), about 1-10% w/w of microcrystalline
cellulose and
colloidal silicon dioxide (e.g., ProsoIv SMCC HD90), about 0.1 - 1% w/w/
magnesium
stearate (e.g. HyQual0), and about 1 - 5% of an enteric coating (e.g. Opadry
1.1 White). in
some embodiments, the method of treating or preventing sialic acid
deficiencies comprises
administering an extended release formulation comprising a drug load of about
30-60% (e.g.,
sialic acid and/or ManNAc), about 20-30% w/w polyethylene oxide (e.g. Polyox
WSR),
about 20-25% w/w sodium alginate (e.g. Protanal), about 1-5% w/w lambda
carrageenan (e.g.
Viscatin GP-209), about 1-10% wlw of microcrystalline cellulose and colloidal
silicon
dioxide (e.g., Prosolv SMCC HD90), about 0.1 - 1% w/w/ magnesium stearate
(e.g.
HyQual0), and about 1-5% of an enteric coating (e.g. Opacity 1.1 White).
[00178] Provided herein are also methods of increasing production of sialic
acid (e.g.,
increasing production of sialic acid in muscle tissue) and the proximate
substrate for
glycosylation, CMP-sialic acid in an individual in need thereof by
administering an effective
amount of one or more compounds in the sialic acid pathway or derivatives
thereof in any
extended release formulation described herein. In some embodiments, the one or
more
compounds in the sialic acid pathway or derivatives thereof include ManNAc or
a derivative
thereof and/or sialic acid or a derivative thereof. For example, provided
herein are methods of
increasing production of sialic acid (e.g., increasing production of sialic
acid in muscle tissue)
in an individual in need thereof by administering an effective amount of
ManNAc and sialic
acid in any extended release formulation described herein.
[00179] Provided herein are also methods of increasing si.alylation of muscle
tissue in an
individual in need thereof by administering an effective amount of one or more
compounds in
the sialic acid pathway or derivatives thereof in any extended release
formulation described
herein. In some embodiments, the one or more compounds in the sialic acid
pathway or
derivatives thereof include ManNA.c or a derivative thereof and/or sialic acid
or a derivative
thereof. For example, provided herein are methods of increasing sialylation of
muscle tissue
in an individual in need thereof by administering an effective amount of
ManNAc and sialic
acid in any extended release formulation described herein.
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[00180] Provided herein are also methods of improving muscle function in an
individual in
need thereof by administering an effective amount of one or more compounds in
the sialic
acid pathway or derivatives thereof in any extended release formulation
described herein. In
some embodiments, the one or more compounds in the sialic acid pathway or
derivatives
thereof include ManNAc or a derivative thereof and/or sialic acid or a
derivative thereof. For
example, provided herein are methods of improving muscle function in an
individual in need
thereof by administering an effective amount of ManNAc and sialic acid in any
extended
release formulation described herein.
[00181.] In some embodiments, conditions associated with one or more genetic
defects in
the sialic acid pathway can also be addressed by treatment with one or more
compounds in
the sialic acid biosynthetic pathway or derivative thereof in any extended
release formulation
described herein, whether presently known or to be discovered. As depicted in
Figure I,
uridine diphospho-N-acetylglucasamine-2-epimerase (UDP-GIcNAc-2-epimerase)
converts
UDP-GIcNAc to N-acetylmannosamine (ManNA.c), which is phosphorylated by ManNAc
kinase in the presence of ATP to produce N-acetylmannosamine-6-phosphate
(ManNAc-6-P).
ManNAc-6-P is converted to N-acetylneuraminic acid-9-phosphate (NeuA.c-9-P)
via
Neu5Ac-9-phosphate synthetase, followed by dephosphorylation of NeuAc-9-P by
Neu5Ac-
9-phosphate phosphatase to yield Neu5Ac (sialic acid). Sial.ic acid then
enters the nucleus
and is converted to cytidine monophosphate-sialic acid (CMP-sialic acid) via
CMP-Neu5Ac
synthetase. In some embodiments, any genetic deficiency regarding ManNAc
kinase,
Neu5Ac-9-phosphate synthetase, or Neu5Ac-9-phosphate phosphatase, or
combination
thereat or condition related thereto, can be treated with an effective amount
of one or more
compounds in the sialic acid biosynthetic pathway or a derivative thereof in
any extended
release formulation described herein. For example, in some embodiments,
administration of
such a compound to block a particular enzymatic step in the pathway yields
treatment of a
condition associated with a defect regarding that particular enzyme.
Accordingly, provided
herein are methods of treating a subject having a condition associated with a
genetic defect
regarding an enzyme in the sialic acid pathway, such as ManNAc kinase, Neu5Ac-
9-
phosphate synthetase, or Neu5Ac-9-phosphate phosphatase or a combination
thereat
comprising administering to the subject an effective amount of compound in the
sialic acid
biosynthetic pathway or a derivative thereof in any extended release
formulation described
herein.
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[00182] In some embodiments, conditions associated with defects in the sialic
acid
biosynthetic pathway that may be treated with a compound in the sialic acid
biosynthetic
pathway or a derivative thereof in any extended release formulation described
herein include,
but are not limited to, sialuria, glomerular hyposialylation, glomerular
hematuria, proteinuria
podocytopathy, renal disorders involving proteinuria and hematuria due to
podocytopathy
and/or segmental splitting of the glomerular basement membrane, minimal change
nephrosis,
focal and segmental glomerul.osclerosis, membranous glomendonephritis,
idiopathic
nephrotic syndrome, and glycosylation deficiencies (e.g., congenital disorders
of
glycosylation or muscular dystrophies). In some embodiments, a condition
associated with a
ManNac kinase defect is selected from sialuria, glomerular hyposialylation,
glomerular
hem.aturia, proteinuria podocytopathy, renal disorders involving proteinuria
and hem.at-uria
due to podocytopathy and/or segmental splitting of the glomerular basement
membrane,
minimal change nephrosis, focal and segmental glomentlosclerosis, membranous
glomerulonephritis, and idiopathic nephrotic syndrome. In some embodiments, a
condition
associated with a Neu5Ac-9-phosphate phosphatase defect is a glycosylation
deficiency (e.g.,
congenital disorders of glycosylation or muscular dystrophies).
[00183] In some embodiments, a cause of sialic acid deficiency, reduced sialic
acid
production, or reduced sialylation is a genetic defect that affects regulation
of the sialic acid
pathway: that is, genetic defects that affect sialic acid productivity or
sialylation need not be
constrained to genetic defects of enzymes directly in the sialic acid pathway.
A.s such, in
some embodiments, any underlying genetic defect involved in regulation of the
sialic acid
pathway that affects sialic acid production (e.g., where the defect causes a
decrease in sialic
acid production or otherwise causes sialic acid deficiency) or sialylation
(e.g., where the
defect causes decreased sialylation) can be treated with an effective amount
of a compound in
the sialic acid biosynthetic pathway or a derivative thereof in any extended
release
formulation described herein. In some embodiments, a genetic defect that
reduces sialic acid
productivity or reduces sialylation, such as by affecting regulation of
GNE/MNK or other
aspects of the sialic acid pathway, can be treated with a compound in the
sialic acid
biosynthetic pathway or a derivative thereof in any extended release
formulation described
herein. Accordingly, provided herein are methods of treating a subject having
a condition
associated with a genetic defect that reduces sialic acid productivity or
reduces sialylation,
such as wherein the defect affects regulation of GNE/MNK or another aspect of
the sialic
acid pathway, comprising administering to the subject an effective amount of a
compound in
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the sialic acid biosynthetic pathway or a derivative thereof in any extended
release
formulation described herein.
[001841 Sialic acids are important for proper development and functioning of
many organs
and tissues, and a deficiency of sialic acid can give rise to many different
types of diseases
and conditions. Other types of muscle diseases have also shown that
glycosylation is
important for muscle function. Nishino and Ozawa, Curr. Op/n. Neurol. 15:539-
544 (2002).
In some embodiments, the sialic acid deficiency is a myopathy, muscular
atrophy and/or
muscular dystrophy. Myopathies that can be treated with the present
compositions and
methods also include distal myopathy with rimmed vacuoles (Nonaka myopathy)
and the
muscular dystrophy hereditary inclusion body myopathy (HIBM). In some
embodiments, the
methods of treating and/or preventing increase sial.ylation of muscle tissue.
In some
embodiments, the methods of treating and/or preventing improve muscle function
and reduce
muscle injury from physical activity, as measures by crearine kinase plasma
levels after
exercise. In some embodiments, the methods of treating or preventing muscle
dysfunction
will improve independent ambulation, stair climbing, foot drop, getting up
from a chair and
walking, hand grip and manipulation and pulmonary function. in some
embodiments, the
method further comprises identifying an individual in need thereof by
determining genotype
or expression levels of the gene GNE.
[00185] In some embodiments, the sialic acid deficiency is a kidney condition
and diseases
(e.g., those involving proteinuria and hematuria). Proteinuria involves
leakage of protein
from the blood into the urine. If the amount of protein in the urine is very
high, this condition
is often called nephrotic syndrome. Several types of diseases exhibit the
symptoms of
proteininia, including high blood pressure, infections, reflux nephropathy,
diabetes, and
various types of glomerulonephritis, including minimal change nephrosis.
Hematuria simply
means blood in the urine (e.g., gross hematuria or microscopic hematuria). in
some
embodiments, the methods of treating and/or preventing increase sialylation of
kidney tissue.
[00186] in embodiments of any of the methods, a therapeutically effective
amount of one
or more compounds in the sialic acid pathway or derivatives thereof is
provided over a period
of greater than about any of 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20
hours. In some
embodiments, a therapeutically effective amount of one or more compounds in
the sialic acid
pathway or derivatives thereof is provided over a period of greater than about
12 hours or
greater than about 24 hours. In embodiments of any of the methods, a
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
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is provided over a period of between about any of 6-10 hours, 8-12 hours, 10-
16, or 12-20
hours. In embodiments of any of the methods, a therapeutically effective
amount of one or
more compounds in the sialic acid pathway or derivatives thereof is provided
over a period of
about any of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 hours. In
some embodiments, a
therapeutically effective amount of one or more compounds in the sialic acid
pathway or
derivatives thereof is provided over a period of about 12 hours or about 24
hours. In some
embodiments, the therapeutically effective amount is provided to the
bloodstream of the
individual. In some embodiments, the therapeutically effective amount is
provided to muscle
tissue of the individual. In some embodiments, the one or more compounds in
the sialic acid
pathway or derivatives thereof include ManNAc or a derivative thereof and/or
sialic acid or a
derivative thereof. For example, in some embodiments, a therapeutically
effective amount of
ManNAc and/or sialic acid is provided to muscle tissue of the individual over
a period of
between about any of 6-10 hours, 8-12 hours, 10-16, or 12-20 hours.
[00187] in embodiments of any of the methods, the individual in need thereof
is provided
a substantially constant (i.e., without large burst of drug availability and
deficiencies in drug
availability to the blood and/or tissues of interest (e.g., muscle tissue))
therapeutically
effective amount of one or more compounds in the sialic acid pathway or
derivatives thereof
over a period of greater than about any of 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, or 20
hours. In embodiments of any of the methods, the individual in need thereof is
provided a
substantially constant therapeutically effective amount of one or more
compounds in the
sialic acid pathway or derivatives thereof over a period of between about any
of 1-24 hours,
4-24 hours, 6-24 hours, 8-24 hours, or 12-24 hours. In embodiments of any of
the methods,
the individual in need thereof is provided a substantially constant
therapeutically effective
amount of one or more compounds in the sialic acid pathway or derivatives
thereof over a
period of about any of 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20
hours. In some
embodiments, the one or more compounds in the sialic acid pathway or
derivatives thereof
include ManNAc or a derivative thereof and/or sialic acid or a derivative
thereof. For
example, in some embodiments, the individual in need thereof is provided a
substantially
constant therapeutically effective amount of ManNAc and/or sialic acid to
muscle tissue of
the individual over a period of between about any of 6-10 hours, 8-12 hours,
10-16, or 12-20
hours.
[00188] In embodiments of any of the methods, less than about any of 10%, 20%,
30%,
40%, 50%, 60%, or 70% of one or more compounds in the sialic acid pathway or
derivatives
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thereof is excreted from the individual after one hour. In embodiments of any
of the methods,
less than about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of one
or more
compounds in the sialic acid pathway or derivatives thereof is excreted from
the individual
after four hours. In some embodiments, the one or more compounds in the sialic
acid
pathway or derivatives thereof include ManNAc or a derivative thereof and/or
sialic acid or a
derivative thereof In some embodiments, the one or more compounds in the
sialic acid
pathway or derivatives thereof include ManNAc and/or sialic acid.
1001891 In embodiments of any of the methods, the one or more compounds in the
sialic
acid pathway or derivatives thereof are administered to an individual in need
thereof between
about any of 0.1-50 g/day, 0.5-25 g/day, 1-15 g/day, 1-10 g/day, 2-5 g/day,
0.2-25 g/day, 0.3-
12 g/day, 0.4-10 g/day, 0.5-8 giday, and 0.7-6 g/day. In some embodiments, the
one or more
compounds in the sialic acid pathway or derivatives thereof are administered
between about 2
g/day and 5 g/day. In embodiments of any of the methods, the one or more
compounds in the
sialic acid pathway or derivatives thereof are administered to an individual
in need thereof
between about any of 0.01-750 mg/kg, 0.5-500 mg/kg, 1-250 mg/kg, 2.5-100
mg/kg, or 5-50
mg/kg. In some embodiments, the one or more compounds in the sialic acid
pathway or
derivatives thereof are administered to an individual in need thereof between
about 5 mg/kg
and 50 mg/kg. in some embodiments, the one or more compounds in the sialic
acid pathway
or derivatives thereof include ManNAc or a derivative thereof and/or sialic
acid or a
derivative thereof For example, in some embodiments, ManNAc and/or sialic acid
are
administered to an individual in need thereof 'between about 5 mg/kg and 50
mg/kg.
[00190] In embodiments of any of the methods, the one or more compounds in the
sialic
acid pathway or derivatives thereof are administered to an individual in need
thereof 'between
about any of 0.01-750 mg/kg/day, 0.5-500 mg/kg/day, 1-250 mg/kg/day, 2.5-100
mg/kg/day,
or 5-50 mg/kg/day, In some embodiments, the one or more compounds in the
sialic acid
pathway or derivatives thereof are administered to an individual in need
thereof between
about 5 mg/kg/day and 50 mg/kg/day. in some embodiments, the one or more
compounds in
the sialic acid pathway or derivatives thereof include ManNAc or a derivative
thereof and/or
sialic acid or a derivative thereof For example, in some embodiments, ManNAc
and/or sialic
acid are administered to an individual in need thereof between about 5
mg/kg/day and 50
mg/kg/day.
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1001911 In some embodiments, the effective amount of one or more compounds in
the
sialic acid pathway or derivatives thereof in any extended release formulation
is administered
once a day, twice a day, three times a day, or four times a day.
[00192] The amount of the extended release formulation according to an
embodiment of
the invention to be administered to a human body may be appropriately selected
in
accordance with the absorption rate in the body, rate of inactivation, rate of
excretion, the
age, gender and condition of the patient, severity of the disease, or the
like. Such factors can
be readily determined by the clinician employing animal models or other test
systems that are
available in the art.
[00193] Administration of the therapeutic agents in accordance with the
present invention
may be in a single dose, in multiple doses, in a continuous or intermittent
manner, depending,
for example, upon the recipient's physiological condition, whether the purpose
of the
administration is therapeutic or prophylactic, and other factors known to
skilled practitioners.
The administration of one or more compounds in the sialic acid pathway or
derivatives
thereof may be essentially continuous over a pre-selected period of time or
may be in a series
of spaced doses. Both local and systemic administration is contemplated.
[00194] In one embodiment, the present invention provides a method for
treating a sialic
acid deficiency in an individual in need thereof comprising administering to a
patient under
fed conditions the present extended release formulation, wherein the extended
release
formulation provides an improved absorption profile when administered under
fed conditions
than being administered under fasting conditions. In some embodiments, the
absorption
profile is described by the area under the plasma concentration-time curve
(A.UC) over a 8,
12, or 24 hours period of time (correlating to the amount of drug absorbed or
bioavailability),
C. (maximum concentration of the drug in the blood), and Tmax (time to reach
C.). In one
embodiment, the improved absorption profile denotes a less sharp and more flat
shape of the
concentration-time curve. For example, the improved absorption profile can
denote an AUC
with somewhat lower, somewhat higher, or substantially the same value but a
lower C. and
higher T.. In one embodiment, the mean AUC determined at a fed state is
substantially
similar to or higher than the mean AUC determined at a fasted state, the mean
C.
determined at a fed state is lower than the mean AUC determined at a fasted
state. In another
embodiment, the mean C. determined at a fed state is lower than the mean A.UC
determined at a fasted state. In another embodiment, the mean T. determined at
a fed state
is higher than the mean AUC determined at a fasted state.
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Unit Dosages and Articles of Manufacture
[00195] Also provided herein are articles of manufacture and unit dosages
which include
the extended release formulations comprising one or more compounds in the
sialic acid
pathway or derivatives thereof described herein.
[00196] Provided herein are articles of manufacture or kits comprising: (a) a
container
comprising the extended release pharmaceutical formulation comprising one or
more
compounds in the sialic acid biosynthetic pathway or derivative thereof
described herein; and
(b) a package insert with instructions for treating and/or preventing a sialic
acid deficiency in
a patient. In some embodiments, the one or more compounds in the sialic acid
pathway or
derivatives thereof include ManNA.c or a derivative thereof and/or sialic acid
or a derivative
thereof.
[00197] The article of manufacture comprises a container and a label or
package insert on
or associated with the container. Suitable containers include, for example,
bottles, vials,
syringes, etc. The containers may be formed from a variety of materials such
as glass or
plastic. The container holds or contains a formulation and may have a sterile
access port (for
example the container may be an intravenous solution bag or a vial having a
stopper
pierceable by a hypodermic injection needle). .At least one active agent in
the composition is
the polypeptide. The label or package insert indicates that the composition's
use in a subject
with specific guidance regarding dosing amounts and intervals of polypeptide
and any other
drug being provided. The article of manufacture may further include other
materials desirable
from a commercial and user standpoint, including other buffers, diluents,
filters, needles, and
syringes. In some embodiments, the container is a syringe. In some
embodiments, the syringe
is further contained within an injection device. In some embodiments, the
injection device is
an autoinjector.
[00198] A "package insert" is used to refer to instructions customarily
included in
commercial packages of therapeutic products, that contain information about
the indications,
usage, dosage, administration, contraindications, other therapeutic products
to be combined
with the packaged product, and/or warnings concerning the use of such
therapeutic products.
[00199] Provided herein are also unit dosages which include the extended
release
formulations comprising one or more compounds in the sialic acid pathway or
derivatives
thereof. In some embodiments, the one or more compounds in the sialic acid
pathway or
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derivatives thereof include ManNAc or a derivative thereof and/or sialic acid
or a derivative
thereof.
[00200] Unit dosage forms comprising any of the extended release formulations
described
herein, including but not limited to those formulations detailed under the
heading "Extended
Release Formulations," such as any of the formulations of Tables A-E, Example
6, or
Example 7, are described. These unit dosage forms can be stored in a suitable
packaging in
single or multiple unit dosages and may also be further sterilized and sealed.
For
convenience and ease of patient compliance, the extended release formulations
may be
delivered in the form. of unit dosage forms, which may be administered to an
individual. In
one variation, the extended release formulation is a solid substance and unit
dosage forms
thereof may be prepared in the form of tablets, capsules, sachets and chewable
tablets or
tablets not intended to be chewed. In one aspect, the dosage form is in the
form of a capsule
or tablet, preferably in the form of a tablet. In some embodiments, the dosage
form is in the
form. of a tablet not intended to be chewed. In some embodiments, the dosage
form is in the
form of a tablet not intended to be crushed. In some embodiments, the dosage
form is in the
form of a tablet not intended to be chewed or crushed.
[00201] The preparation of the unit forms generally involves a step of
preparing a blend
filling, either by volume or weight. For example, in production of tablets and
capsules, the
extended release formulation blend is volume filled into a die or capsule,
respectively. In one
aspect, a batch of unit dosage forms has the same potency (amount of drug per
unit dosage
form) within an allowable margin, which in one variation is a relative
standard deviation
(R.SD) of less than 6% and in another variation is less than 8.0 or 7.8%.
EXAMPLES
[00202] The following examples are put forth so as to provide those of
ordinary skill in the
art with a complete disclosure and description of how to make and use the
present invention,
and are not intended to limit the scope of what the inventors regard as their
invention nor are
they intended to represent that the experiments below are all or the only
experiments
performed.
Example 1
Preparation of Sial.ic Acid 250 mg Strength Tablets Using Dry Blend Method of
Manufacture
Experimental/Materials
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1002031 Sialic Acid (Food & Bio Research center, Inc. Kyoto Japan) was stored
in
aluminum foil bags at -20 C. However, handling and processing of prototypes
were all under
ambient room temperature. In-process materials and bulk tablets were stored in
double
polyethylene bags with desiccant. The siatic acid was evaluated for physical
properties
consisting of morphology, particle size by sieve analysis, bulk and tap
density.
100204] 50 gram lab-scale batches were prepared using bag-blending, manual
filling and
hand turning of the tablet press to compress tablets to evaluate dissolution
as the first level of
screening. Tablets were manufactured using the ProCR platform, Their formulas
are listed
below in Table I and 2.
Table I. Quantitative Formula for Sialic Acid, ProCR Hypromellose 250 mg
Tablets:
N
Food arid
Sialie Acid 0\T-
BioResearch Center, 249.75 33.3 16.65
Acetylnet [ram inic acid)
Inc
Hypromellose, Type 2208
(Methocel K100 M Colorcon 225.0 30.0 15.0
Premium CR)
Sodium Alginate
FMC Biopolymer 1187.5 25.0 17.5
(Protanalci'' 1_,F 120M)
Carrageenan (Viscarin GP-
FM(' Biopolymer 37,5 7.5
209)
Microcrystallline Cellulose
and Colloidal Sillicon
JRS Pharma 46.5 6.2 3.1
Dioxide ( ProSolv SM CC
HD 90)
Magnesium Stc.arate
(HyQuar), Vegatable Mallinckrodt 3.75 0.5 025
Source Product Code 2257
Total 750 100% 50
Table 2. Quantitative Formula for Sialic Acid ProCR Polyox, 250 mg Tablets:
lajgredletit !:Veadf.jt: A'Atgi tablet
::%W* gibatell0
Food and
Sialic Acid(1\1-
BioResearch Center, 249.75 33.3 16.65
Acetylneuraminic acid)
Inc
Polyethylene Oxide WSR Dow Chemical
775.0 30,0 15.0
(Polyox) Company
Sodium Alginate
FMC Biopolymer 187.5 25.0 12.5
(Protanal, LF 120M)
Carrageenan (Viscarin GP-
FMC Biopolymer 37.5 5.0 2.5
209)
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Microcrystalline Cellulose
and Colloidal Sillicon
.111S Pharma 46.5 6.2 3.1
Dioxide (1?roSolv S M CC
HD 9O)
Magnesium Stearate
(HyQual ), Vegatable Mallinckrodt 3.75 0.5 0.25
Source Product Code 2257
Total 750 100 50
[00205] Sialic Acid, hyprom.ellose Type 2208, sodium alginate, carrageenan and
microcrystalline cellulose with colloidal silicon dioxide were delumped using
a #20 USA
standard sieve and weighed per the quantitative formula. The ingredients were
combined in a
small ziplock bag and blended for 15 minutes. Magnesium stearate was delumped
using a #
40 USA standard screen, weighed per quantitative formula, and added to the
blended
ingredients in the bag. The ingredients were blended for an additional three
minutes. The
fmal blends, as well as the un-sieved sialic acid were characterized using
bulk density, tap
density, particle size sieve analysis, Can's Compressibility Index, and
minimum critical
orifice. The final blend of each prototype was compressed on the Korsch PH100
tablet press.
The resulting tablets were submitted to the analytical lab for dissolution
testing.
Sialic Acid Characterization
[00206] Sialic Acid was visually characterized as a white fluffy powdery
substance. Its
bulk density was 0.293g/m1,, and its tap density was 0.419 g/ml. The Can's
Compressibility
Index was 30 %, and the minimum critical orifice diameter was 18 mm. The
particle size
sieve analysis of Sialic Acid (Table 3) revealed a distribution of coarse and
midsize particles
as shown in Figure 2. The sialic acid was sized prior to blending to
facilitate blend
homogeneity.
Table 3. Particle Size Distribution for Sialic Acid
Sieve #( Mesh size (um)) Unsieved Sialic Acid (N-
Acetylneuraminic acid)
20(850) 34.16
40 (425) 26.87
60 (250) 15.57
100 (150) 10.75
200 (75) 9.3
325 (45) 1.77
Pan (<45) 1.45
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ProCR Sialic Acid, 250 mg CR Tablets
[00207] Both prototype blends, ProCR hypromellose and ProCR Polyox, were
compressed
into tablets using 0.3300 X 0.7100 inch modified oval tooling targeting a
tablet weight of 750
mg and a hardness range of 17 to 20 Kp. During tableting, powder bridging in
the die cavity
was observed for ProCR hypromellose. This was an indication that the blend
needed to be
densified to improve &mobility on the tablet press. ProCR Polyox appeared
denser and
seemed to flow better on the tablet press. However, its Can's Compressibility
Index and
minimum critical orifice diameter results, as shown in Table 4, indicated that
it also needed
further processing such as, granulation. The particle size distribution of the
Polyox prototype
seemed to be more dispersed over various screen sizes than the hypromellose
prototype
shown in Table 5 and Figure 3.
Table 4. Physical Characterization Results of Sialic Acid 250 mg
Carr's :]]Flodex
Bulk Density Tap Densityg
Powder :]]]]]g:m
Compressibility miMCritical
(g/mL) (g/mL)
Index (%) Oiifiee (mm)
_________________________________________ =
Sialic Acid (N-
Acetylneuraminic 0.293 0.419 30 (poor flow) 18
acid)
Prototype 1 0 359 0 543 33.8 (very poor
..
(Hypromellose) flow)
Prototype 2 38.7 (very very
0439 0716 18
(Po I yox) . . poor flow)
Table 5. Particle Size Distribution for :ProCR Sialic Acid, 250 mg Tablets.
Pittotype 1 with Petittitype 2 With"
Sieve #( Mesh size (um)) J1ypromeilose
Polyethylene oxide
% Retain % Retain
20 (850) 0.34 0.84
40 (425) 16.02 15.20
60 (250) 9.2 12.88
100(150) 10.3 15.49
200 (75) 10.3 18.51
325 (45) 33.91 24.96
Pan (<45) 19.89 12.12
[00208] The compression of the tablets resulted in a weight range of 3- 5% of
the target of
750 mg. The variability was primarily due to the manual filling and poor flow.
Regardless of
the weight variability, the tablet appearance and hardness was good, ranging
from 13 to 18
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Kp, as listed in Table 6. The dissolution results showed a first order
sustained release profile
over a 12 hour period, as shown in Table 7 and Figure 4.
Table 6. Physical Data of Sialic Acid 250 and 325 mg Tablets
Hvpromellose Po1)o Rpromel1ose Polvox
-
Test 250 mg 250 mg 325 mg 325 mg
Tablets Tablets Tablets
Tablets
Tablet Weight (mg) 744 - 787 746 - 751 747 -
766 745 - 771
_ Tablet Thickness (in) 0.268
¨ 0.271 0.261 ¨ 0.263 0.291 ¨ 0.295 0.283 -0.286
Tablet Hardness (kp 17.5 18.3 13.2 13.0
Tablet Friability (%) NI) ND ND ND
ND: Not determined
Table 7. Dissolution Results of Direct Compression Prototypes
11.ypromellose Polvox llypromellosei Polyox
Test 250 mg 250 mg 325 mg 325 mg
Tablets Tablets Tablets
Tablets
Dissolution (Average n=3)
% Release
2 lir 26 30 33 35
4 hr 41 50 50 55
6 hr 53 66 67 71
8 hr 65 77 78 82
12 hr 82 97 92 95
16 hr * 99 103
24 hr 100 j 112
* Represented as last time point in graph
Example 2
Preparation of Sialic Acid 325 and 500 mg Development Prototypes
[00209] Initially, two small 50 gram dry blend batches were manufactured with
an
increased drug load from 33%w/w to 43%w/w to verify that the drug release
profile was
acceptable. The two compositions are listed in Table 8 as hypromellose and
Polyox. The
tabletting was done as described before using a manual fill into the die
cavity.
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Table 8. Quantitative Formula for Sialic Acid 325 mg and 500 mg sustained
release
Tablets Prototypes:
mg/Tablet mg/Tablet batch Whatch
g/
Ingredient Vendor ProCR ProCIR %wiw
1.800 g
50 g size
Hypromellose Polyox size
Sialic Acid (N- Food and
.Acetylneuraminic BioResearch 325.0 325.0 43.3 21.65 779.4
acid) Center, Inc
Hypromellose,
Type 2208
(Methocel 1(100 Colorcon 191.3 25.5 12.75 459
M Premium CR)
Polyethylene Oxide
191.3 25.5 12.75 459
WSR (Polyox)
Sodium Alginate
FMC
(Protanar LF159.0 159.0 21.2 10.60 381.6
120M)
Biopolymer
Carrageenan FMC
31.5 31.5 4.2 2.10 75.6
(Viscarin GP-209) Biopolymer
Microcrystal.11ine
Cellulose and
Colloidal Silicon IRS Pharma 39.8 39.8 5.3 2.65 95.4
Dioxide (ProSolv
SMCC HD 90)
Magnesium
Stearate
(HyQualc), Mallinckrodt 3.8 3.8 0.5 0.25 9.0
Vegatable Source
Product Code 2257
Total for 325mg Strength 750.4 750.4 100 50 1800
Total for 500 mg Strength 1154.5 1154.8 100
Wet Granulation Method of Manufacture
[00210] in order to avoid bridging and poor flow during compression batch
sizes were
scaled .up from 50 grams to 1800 grams, and a high shear granulation method of
manufacture
was used to produce 325 and 500 mg dose strengths while maintaining good
tablet
compression properties. The 325 and 500 mg dose strengths shared a common
blend that was
divided prior to compression. Two tablet sizes were produced: A 325 mg dose
strength tablet
with a length of 17.7 mm, a width of 9.1 mm and a thickness of 6.7 mm; and a
500 mg dose
strength tablet with a length of 19.3 mm, a width of 9.7 mm and a thickness of
8.0 mm). The
following equipment and process were used to make these tables.
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Experimental/Materials
[00211] All raw materials were used as received from vendors as listed in
Table 8. The
batch size was 1800 grams. The following equipment was used:
Fielder PP1 High Shear Granulator
N iro-Aeromatie MP-1 Multi-processor
Homoloid equipped with knives forward, 0.079" round hole screen
4 Qt PK Blender
Korsch PH100 tablet Press equipped with 0.350" x 0.6875" modified oval tooling
for the 750 mg tablet and 0.374" x 0.7480" modified oval tooling for the 1154
mg
tablet
Accela-cota model 24MK III (24" coating pan)
[00212] All the raw materials with exception of magnesium stearate were
charged to the
PP-1 granulator and premixed for 3 minutes at 300 rpm impeller speed, no
chopper. A
baseline loss on drying determination was performed and the ungranulated
hypromenose
formula was determined to be 3.4% water white the Polyox formulation was 2.9%.
Water
was sprayed at approximately 200 grams/minute while mixing at 300rpin with a
slow
chopper speed. The hypromenose formulation used 43% water (778g water sprayed)
of the
1.8kg batch size while the Polyox formulation sprayed 52% water (905g water
sprayed) with
a 2 minute post spray mix. The granulation was transferred into the MP-1 fluid
bed and dried
with an inlet temperature of 75 C until the loss on drying (LOD) was < 3%;
equal to or
slightly lower than the baseline moisture of the un-granulated formulations.
The dried
granulation was passed through a #4 mesh hand screen. The large granules
retained on the #4
mesh were segregated and discarded. The remaining granules were sized through
the FitzMill
at low speed, knives forward. The blend was then lubricated with the magnesium
stearate for
3 minutes. The final blend was compressed into tablets using a Korsch rotary
press. After
dissolution results were obtained, the core tablets were coated with a non-
functional, Opadry
11, white to a weight gain of approximately 4.5%w/w.
100213] Outline of Dissolution Conditions were as follows:
[00214] 900 int dissolution medium: 50mN4 Phosphate, pH 6.8
100 RPM Baskets
37 C
Time points: 2, 4, 6, 8, 12, 16 or 24 hours
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[00215] The blending and granulation of the hypromellose based formulation
proceeded
smoothly. The hypromellose formulation processed well, producing a final blend
with
excellent flow that compressed well on the tablet press. The yield was
excellent (96%) for a
small scale batch size.
[00216] The Poly Ethylene Oxide (Polyox) based formulation did not granulate
as easily.
The Polyox formulation was over-granulated. The over-granulation can be
alleviated in the
future by spraying less granulation water at a slower rate. An appreciable
amount of the
granulation was lost when the partially dried granulation was screened through
a 4 mesh
sieved to remove large over-granulated agglomerates that resisted drying in
the fluid bed. As
a result, the batch yield was poor at 83%. The portion of the batch that was
retained
produced an excellent final blend, however. It flowed and compressed well on
the tablet
process and produced good quality tablets. Polyox is known for being difficult
to granulate so
this is not entirely unexpected. However, with the proper granulation
parameters an excellent
granulation can be attained.
[00217] Physical data for sialic acid 325 mg final blends, sialic acid 325 mg
tablets, and
sialic acid 500 mg tablets are shown in Tables 9, 10 and 11, respectively.
Analytical results
for sialic acid 325 and 500 mg tablets (uncoated) are shown in Table 12.
Table 9. Physical data for Sialic Acid, 325 mg Final Blends:
___________________________________________________ (proCR H)pomellose)
(ProC11 Polyox)
Sieve # (% Retain)
Mesh size (um)
14(1400) 0.10 1.32
30(600) 42.89 45.4
40 (425) 12.28 14.39
140 (106) 33.98 29.89
200 (75) 5.42 3.24
325 (45) 4.61 4.86
Pan (<45) 0.72 0.91
Blend Bulk Density (g/rnL) 0.54.9 0.54.5
Tap Density (g/mL) 0.646 0.619
% Compressibility 15 12
Flowdex 10 6
Table 10. Physical Data of Sialic Acid 325mg Tablets at Various Ilardnesses
Formula Formula Formula Formula Formula Formula
Test .
________________________________________ floe A _ tion B tion C tion D
tion E tion F
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1 Hypromellose
Tablet Hardne High / High i
Low Medium Low Medium
Level max
max
Ave. Weight (mg) 1 759.5 754 754 739 745 750
Ave. Thicluiess (in) 0.279 0.270 0.259 0.260
0.247 0.253
Ave. Hardness (kp) 6.5 10.0 14.4 ------- 9.5 17.9 15.7
Ave. Friability (%) Failed 0.2 s 0.1 0.1 0.0
0.2
Note: Average of 10 tablets
Table 11. Physical Data of Sialic Acid 500 mg Final Blends and Tablets
Formula Formula ' Formula FOrm u la ' Formula - Formula
I k.st thin G
ton El tion 1 tion 3 tion K
lion L
. _ - ¨ .
ilypromellose Pot) ox
Bulk DL:t;siiy (gluil.) 0.55 0.54
Tablet Hardness Medium High /High /
Low Low Medium
Level * maxmax
Ave. Weight (mg) 1170 ND 1152 1158 1154 1160
Ave. Thickness (in) 0.324 ND 0.315 0.310 0.307 0.297
Ave. Hardness (kp) 11.3 ND 13.2 12.9 14.0 20.2
Ave. Friability (%) 0.2 ND 0.0 0.1 0.0 0.0
Note: Average of 10 tablets
* ND: not determined
Table 12. Analytical Results or Sialic Acid 325mg and 500mg SR Tablets
(Uncoated)
Formulation Formulation Formulation Formulation
Test C F I K .,
11:3promellose Polyox Ilypromellose Polyox
500 500
lablet Strength (mg) 325 325
.__
Assay (%LC) 96.3 97.8
Impurities (%) Total: 0.2 Total: 0.2
-- 7
Dissolution (Average n=3) 1
% Release
2 hr 34 34 29 29
4 hr 53 55 46 47
6 hr 67 72 59 62
8 hr 78 85 69 74
12 hr 91 96 84 89
24 hr 100 99 101 1 100
1002181 The dissolution results (Figure 5) showed a first order sustained
release profile
over a 12 hour period for both dose strengths and for both ProCR hypromellose
and ProCR
Polyox. Additionally, these results indicate that the dose proportional
approach was
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successful in providing dose flexibility using a common blend at 750 and 1154
tablet final
weights.
Example 3
Coating for Sialic Acid 325 and 500 mg SR Tablets ProCR Hypromellose and ProCR
Polyox
Method of Manufacture
100219] Eight kilograms of core tablets (approximately 1.5kg of active tablets
combined
with 6.5 kg of "sham" placebos to provided volume) were charged into an Accela-
Cota
coating equipment equipped with a 24" coating pan and two spray guns. The non-
functional
film coat was Opadry-II White (Colorcon Corporation formula Y-22-7719) at a
20% solids
concentration, The purpose of the film coat was to improve aesthetics and in
the future
facilitate patient compliance for swallowing of the tablet. The target end-
point was 3-5%
weight gain.
[00220] The coating process parameters were as follows:
Pan speed: Target 12 -16 rpm
inlet temperature: 70-85 C
Outlet temperature: 39-42 C
Bed temperature: 33-45 C
Atomization pressure: 40 psi
Spray Rate: 50 - 60 glinin
Airflow: approximately 200 cfm
Gun to bed distance: 5"
[002211 The tablets coated well with no difficulties. Approximately 4% weight
gain of
coating was sufficient to provide good coverage of the tablet cores.
Prototype Stability
[00222] The white film coated tablets of Sialic Acid prepared using ProCR.
hypromellose
and ProCR Polyox at 325mg and 500mg dose strengths were packaged in thirty
(30) units per
bottle, one MiniPax desiccant, no coil and induction sealed using a Lepak Jr
TM induction cap
sealing system. Table 13 lists the packaging components used. All the
acceptable tablets were
packaged and placed on a 12 month prototype stability program under ICH
conditions testing
the stability at both 25 C and 60% relative humidity (RH) and 40 C and 75% REI
at 0, 1, 3, 6,
and 12 months. The tablets have been tested and monitored with respect to
appearance,
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dissolution, moisture, assay and related substances, and initial stability
results are shown in
Table 14. The dissolution profile for the coated 325mg and 500 mg tablets is
shown in Figure
6.
Table 13. List of Packaging Components
Component Material Description Al RM #
Bottle 100cc Round White HDPE (38/400) PC-3714
Closure 38mm CRC w Foil Seal MI Liner PC-3982
Desiccant MiniPax w 1.00g Silica Gel-Packet PC-2637
Table 14. Analytical Results of Sialic Acid 325mg and 500mg SR Tablets
(Coated),
Initial Stability
Formulation Formulation Formulation Formulation
Test C F I K
Hyprom el lose Po lox Hypromeiiose Po lyox
Tablet Strength (mg) 325 325 500 500
% Moisture by Karl Fischer 1.0 3.3 2.0 3.7
Content Uniformity
(Ave, n..:10) 100.0 95.6 99.8 98.6
%RSD 1.5 2.5 1.5 2.6
AV 3.5 9.2 3.6 6.3
Assay (%LC) 100.6 97.8 98.8 96.9
Impurities (%) Total: <0.10 Total: <0.10 Total: <0.10 Total: <0.10
Dissolution (Ave. % Release,
n 6))
2 hr 31 34 27 27
4 hr 50 54 42 43
6 hr 65 70 55 56
8 hr 75 83 65 68
12 hr 90 96 79 83
24 hr 102 100 98 96
[00223] The formulation development activities successfully identified two
distinct
sustained release prototypes for Sialic Acid in 325 and 500 mg dose strengths.
The in-vitro
dissolution release profile exhibited a first order release over 12 hours in
aqueous medium
and pH of 6.8. The sustained release ProCR platform was employed. This unique
combination of inert polymers provides a robust formulation that is pH
independent and lends
itself to granulation processes without affecting the dissolution release
profile. This was the
case for Sialic Acid 325 and 500 mg dose strength SR tablets where a wet
granulation process
was found necessary to achieve densification and good tablet compressibility.
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1002241 With regard to chemical stability Sialic Acid 325 and 500 mg ProCR
hypromc.dlose and ProCR Polyox SR. tablets showed acceptable assay,
dissolution and content
uniformity and easily passed TISP testing criteria. These prototypes are
monitored through a
12 month ICH stability study.
1002251 As shown in Figures 5 and 6, the dissolution profiles of Sialic
Acid ProCR
hypromellose and ProCR Polyox uncoated and coated tablets are consistent.
There is no
significant change in the sustained release profile over the 12 hour release
with the
application of Opdary II White film coat. The analytical results for assay
and related
substances are acceptable which indicates that the wet granulation, drying and
coating
processes have no impact on the chemical integrity of the drug.
Example 4
Preparation of ManNAc 325 mg Development Prototypes
The ManNAc title tbi ___ mutation was prepared according to the method
detailed
above for Sialic Acid. The dissolution profile of ManNAc 325 mg Tablets is
shown in Figure
7.
Core Tablet Results
Assay
%LC = 93.5%
Impurity %RS
Sialic Acid <0.10%
Sodium Pyruvate < 0 10%
N-Acetyl-D-Glucosamine 0.4%
Acetic Acid < 0.10%
'Total 0.4%
KF
Pre ) % water
1.5
3.3
Mean'2) 3.4
Content Uniformity
Unit %LC
93.7
94.6
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3 92.8
4 92.8
94.6
6 92.9
7 96.0
8 95.4
9 92.5
91.5
Mean (10) 93.7
%RSD 1.5
SD 1.42735186
AV 8.2
Example 5
Pharm.acokinetics of sialic acid formulations following a single oral or
intravenous dose in
male dogs
[00226] The objective of this study was to evaluate the pharrnacokinetics of
sialic acid
following single oral or intravenous dose in male dogs.
[00227] A total of six male beagle dogs (Canis familiaris), originally from
Beijing
Marshall Biotechnology Co., Ltd., were obtained from the PCS-SHG colony and
subjected to
a general physical examination to ensure normal health status before study
initiation. All
animals were considered suitable for use and each animal was uniquely
identified by a
permanent skin tattoo number andior letter on the ventral aspect of one pinna.
An acclimation
period of five days was allowed between animal transfer and the start of
treatment in order to
accustom the animals to the laboratory environment.
[00228] Before dosing initiation, all animals were weighed and assigned to
treatment
groups. At the start of treatment, animals were 7-16 months of age and ranged
in weight as
6.4 to 9.4 kg. Animals were housed individually in stainless steel cages
equipped with a
mesh-type floor and an automatic watering valve. A standard certified pelleted
commercial
dog food (approximately 400 g of Certified Canine Diet 5C07, PM! Nutrition
International,
Inc.) was provided to each animal once daily, except during designated
procedures.
Maximum allowable concentrations of contaminants in the diet (e.g., heavy
metals, aflatoxin,
organophosphates, chlorinated hydrocarbons, PCBs) were controlled and
routinely analyzed
by the manufacturers. It was considered that there were no known contaminants
in the food
that could have interfered with the objectives of the study. Municipal tap
water, which was
softened, purified by reverse osmosis and exposed to ultraviolet light, was
freely available
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except during designated procedures. It was considered that there were no
known
contaminants in the water that could have interfered with the objectives of
the study. Each
Animal was provided with a floor toy, except during designated activities.
[00229] The study design was as shown in Table 15:
Table 15
Experimental Design
Dose Number Dose Dose
Group Study Lever of Tablets
Volume Concentration Animal
No. Day Test Article Treatment' (mg/kg) per animal (mUkg)
(mg/mL) Number
TA-1
1 PO 10
(capsule)
6 TA-6 IV 25 0.5 50
1 TA-2 3
9 PO 10
(tablet)
TA-3
13 PO 10
(tablet)
TA-1
1. PO 10
(capsule)
6 TA-6 IV 25 0.5 50
2 TA-4 3
9PO
(tablet)
TA-5
13 PO
(tablet) ---------
- ¨ not applicable.
# Dose level was expressed as free form.
a Animals were fasted overnight for approximately 16 hours prior to each dose
and were fed immediately
after the 6 hr timepoint.
[00230] The first day of dosing was designated as Day 1. The subsequent dosing
days
were Days 6, 9 and 13. On Days 1, 9 and 13, all animals were orally
administered prepared
capsules or tablets. On Day 6, all animals received a single intravenous dose
of TA-6 at
0.5 mL/kg. Each actual volume of TA-6 administered was based on the most
recent practical
body weight of each animal. The test articles 1 through 6 are specified in
Table 16.
Table 16
Specification of Test Articles
TestCompositonl
Animal
Identification Dose Level
Article Concentration
Number
TA-1 API in Capsule Form 325 mg SA per capsule 3250
mg SA per 101-103,
animal 201-203
TA-2 Sialic Acid Delayed Release Tablet 325 mg SA / 425 mg excipient 3250 mg
SA per 101-103
Formulation 1 (hypromellose) per tablet animal
TA-3 Sialic Acid Delayed Release Tablet 325 mg SA / 425 mg excipient 3250 mg
SA per 101-103
Formulation 11 (polyethylene oxide) per tablet animal
TA-4 Sialic Acid Delayed Release Tablet 500 mg SA / 650 mg excipient 5000 mg
SA per 201-203
Formulation HI (hypromellose) per tablet animal
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TestCo nip sitonl Animal
Identification Dose Level
Article Concentration
Number
TA-5 Sialic Acid Delayed Release Tablet 500 mg SA/ 650 mg excipient 5000 mg SA
per 201-203
Formulation IV (polyethylene oxide) per tablet animal
TA-6 Sialic Acid IV Formulation 50 mg/mL 25 mg/kg SA 101-
103,
201-203
[00231] Individual body weights were measured once during the predose period
and prior
to each dose on dosing days. There were no treatment-related clinical signs
observed during
the study period and no treatment related changes in body weight or body
weight gains noted
for any animal during the study period.
[00232] Blood samples were collected into serum separate tubes from all
animals on Days
1, 6, 9, 13 for processing to serum at the following time points: predose, 2
minutes (iN, only),
minutes (i.v. only), 10 minutes (i.v. only), 15 minutes, 30 minutes, 1, 2, 4,
6, 8 and 24 hours
postdose. Urine samples were collected into jars on wet ice or ice packs from
all animals on
Days 1, 6, 9, 13 at the following time intervals: predose (overnight for
approximate 15 hours),
0 to 4, 4 to 8, 8 to 12 hours postdose. Samples were collected according to
Table 17 and
Table 18:
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Table 17
PK Sample Collection Schedule
Sample Collection Time Points
Group __________________ (Time Post Dose) on Days 1, 6,9 and 13
No. Omina 2min 5nai 10min 15min 30min 1 hr 2 hr 4 hr 6 hr 8 hr 24 hr
X X
X X(TV only (IV only X X X X X
X X X
(IV only)
X X X
2 X (IV only (IV only X X X X X
X X X
(IV only)
x = sample collected
a Samples were collected before dosing.
Table 18
Urine Sample Collection Schedule
Sample Collection Time Points
Group (Time Post Dose) on Days 1, 6,9 and 13
No. Overnight' 0-4hr 4-81ir 8-12hr
X X X
2 --------- X X X X
x ¨ sample collected
a Blank urine were collected overnight (approximate 15 hours) before each
dose.
[00233] Blood samples were placed at room temperature for at least 30 minutes
but no
more than 1 hour to clot prior to refrigerated centrifugation (approximately 4
C) at
approximately 2700 rpm for 10 minutes. The serum separated from each sample
was
transferred into polypropylene tubes and placed on dry ice until transferred
to a freezer (set to
maintain -80"C). Urine samples were stored in a freezer (set to maintain -80
C) until
analyzed.
002341 Drug concentrations in serum and urine were determined by LC MS/MS
using a
validated analytical procedure (Validation of a Method for the Determination
of Free Soluble
Sialic Acid in Dog Serum and Urine by Liquid Chromatography-Tandem Mass
Spectrometry
(LC-MS/MS) - PN 102653; Long-term Matrix Stability Assessment of Free Soluble
Sialic
Acid in Dog Serum and Urine by Liquid Chromatography-Tandem Mass Spectrom.etry
(LC-
MS/MS) - PN 102654. The method had a linear range from 10 ¨1000 p.g/mL and the
lower
limit of quantitation was 10 ilg/mL.
[00235] Data collection was performed using Analyst from AB Sciex.
Statistical
analyses including regression analysis and descriptive statistics including
arithmetic means
and standard deviations, accuracy and precision were performed using Watson Tm
Laboratory
Information Management System (L1MS) and Microsoft Excel.
[00236] Pharrnacokinetic parameters were estimated using WinNonlie
pharmacokinetic
software (Version 5.2.1, Pharsight Corp., Mountain View, California, USA.).
A
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non-compartmental approach consistent with the intravenous or oral route of
administration
was used for parameter estimation. All parameters were generated from
individual sialic acid
concentrations in serum. Parameters were estimated using nominal sampling
times relative to
the start of each dose administration. Mean concentrations were derived from 3
animals
/group/time point for intravenous dosing occasion only. The actual timepoints
were within
the range of protocol specified. Serum concentration values obtained at the
predose time
point were used as the concentration at time zero for oral doses. The actual
dose levels of test
articles 1 through 5 were calculated using the total amount of SA given to
each animal based
on their most practical body weight.
[00237] The area under the sialic acid individual serum concentration versus
time curve
(AUC) was calculated using the linear trapezoidal method with linear
interpolation. The
terminal elimination phase of each individual concentration versus time curve
was identified
using at least the final three observed concentration values. The slope of the
terminal
elimination phase was determined using log linear regression on the unweighted
concentration data. The terminal elimination phase related parameters were not
reported if
the coefficient of determination was less than 0.800, or the extrapolation of
the AUC to
infmity represented more than 20% of the total area, or the terminal
elimination phase could
not be identified. The parameters described in Table 19 were observed or
calculated.
[00238] All data from serum including concentrations below LLOQ (except for
those
below zero) were applied to pharmacokinetic analysis.
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
Table 19
Estimated Parameters from Serum Concentrations of Sialic Acid
Parameters Description of parameter
Cmax The maximum observed arithmetic individual concentration of
sialic acid after dosing.
Tmax The time after dosing when the maximum observed arithmetic
individual concentration of
sialic acid was observed.
AUC(0-t) The area under the sialic acid arithmetic individual
concentration versus time curve from
time zero to the time after dosing when the last quantifiable concentration of
the drug was
observed.
MRT(0-t) The mean residence time of sialic acid estimated from lime zero
to the time after dosing at
which the last quantifiable concentration of the drug was observed estimated
or imputed by
the linear or linear/log trapezoidal method.
T1/2 The apparent terminal elimination half life.
AUC(0-int) The area under the arithmetic individual concentration versus
time curve from time zero
to infinity.
RT(0-in I) The mean residence time estimated from time zero to infinity.
CL(IV only) Clearance: the apparent volume of serum cleared of sialic acid per
unit time following
intravenous dosing. Clearance was calculated for intravenous dose only.
Vd (IV only) The apparent volume of distribution of sialic acid, determined
from the terminal elimination
phase following intravenous dosing. Volume of distribution was calculated for
intravenous
dose only.
Fs Absolute bioavailability based on sialic acid levels in the serum
following IV and oral
administration.
[00239] Urinary concentrations of sialic acid were subjected to calculation
using
Microsoft Excel, 2007. All data from urine including concentrations below
LLOQ (except
for those below zero) were applied.
[00240] The data of predose urine samples were applied to calculate the total
increase in
urinary excretion of sialic acid at 12 hours postdose. The urinary excretion
of sialic acid, as a
percent of dose administered was estimated for each dosing occasion. Based on
the
assumption that the amount of drug excreted in urine after oral administration
was a
reflection of the dose absorbed, the bioavailability of sialic acid was
determined based on the
percent excretion value following IV and oral administration.
[00241] The parameters described in Table 20 were observed or calculated.
Table 20
Estimated Parameters from Urine Concentrations of Sialic Acid
Parameters Description of parameter
Dose The amount of SA dosed per animal contained in each test
article.
Mass Excreted (0-12hr) The total mass of urinary excretion of sialic acid
at 12 hours postdose.
The corrected value calculated by subtracting out the background masses
Mass Excreted (0-12hr)- based on predose data, representing the increase in
urinary excretion of sialic
Corrected acid at 12 'hours postdose. If corrected value was less
than zero, the value
was set to zero.
Percent Excretion(0- 12hr) The corrected mass excreted of sialic acid as a
percent of dose administered.
Bioavailability based on the sialic acid levels in the urine following IV and
oral administration.
86
CA 02862836 2014-07-02
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[00242] There were no treatment-related clinical signs noted following either
oral or
intravenous administration of sialic acid over the study period. Skin red was
noted for Animal
Nos. 201 and 203 during the study, which was considered as incidental.
[00243] There were no treatment related changes in body weight or body weight
gains
noted for any animal during the study period. Any differences in body weight
or body weight
gain were likely related to expected biological variation.
[00244] Individual concentrations of sialic acid versus time in Beagle dog
serum following
IV or oral administration are shown in Figures 8A-8H.
TA-1
[00245] The background sialic acid levels were below zero for predose samples
of five of
the six animals, except for Animal No. 103, of which was slightly above zero
but below 20%
of the LLOQ.
[00246] Following oral administration of TA-1 in prepared capsules at 3250 mg
of SA per
animal, peak concentrations were observed ranging from 12.6 to 40.8 pg/mL. T.
was
observed at 2 hours postdose with the exception of Animal No. 103 (0.5 hours).
The
concentrations of sialic acid decreased to levels below zero at 24 hours
postdose for all six
animals. The concentration of Animal No. 201 at 24 hour postdose (22.0989
pg/mL) was
considered as aberrant and excluded from analysis, as it was a >LLOQ value but
following
three <LLOQ samples which followed three quantifiable concentrations in the
sampling
sequence.
[00247] Towards the end of the sampling period, a decrease in sialic acid
concentrations
was apparent, but the terminal elimination half-life could only be calculated
for
Animal Nos. 103, 201 and 203, ranging from 1.39 to 1.49 hours.
[00248] The bioavailability of TA-1 was estimated to be ranging from 2.73% to
6.76%,
based on the individual AIJC(04) value following IV and oral administration.
[00249] All the predose urine samples had a measureable concentration of
sialic acid while
the data varied for each individual, ranging from 8.16 to 25.111g/mL. The
maximum
excretion of sialic acid was observed for samples collected 4-8 hours
postdose, except for
Animal No. 202 (0-4 hours postdose). The total mass of sialic acid excreted in
the urine was
equivalent to 0.43-3.56% of the doses of SA contained in TA-1.
[00250] The bioavailability of TA-1 was estimated to be ranging from 1.29% to
39.1%
based on the individual urinary percent excretion value following IV and oral
administration.
87
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
TA-2
[00251] The background sialic acid levels were below zero for predose samples
of
Animal Nos. 102 and 103 except for Animal No. 101, of which was slightly above
zero but
below 20% of the LLOQ.
[00252] Following oral administration of TA-2 in prepared tablets at 3250 mg
of SA per
animal, Tmax was observed from 2.00 to 4.00 hours postdose with the peak
concentrations
ranging from 7.98 to 13.7gg/mL. The concentrations of sialic acid generally
decreased after
T. to levels below zero at 24 hours postdose, for all dosed animals. The
elimination half
life of sialic acid was estimated to be 1.28 hour in Animal No. 103. For
Animal Nos. 101 and
102, the half-life could not be estimated as the measurable data were not
enough to identify
the termination elimination phase.
[00253] The oral bioavailability of TA-2 was estimated to be ranging from
1.64% to
3.25%, based on the individual AUC(0-t) value following IV and oral
administration.
[00254] All the predose urine samples had a measureable concentration of
sialic acid
ranging from 13.5 to 34.8 ILtg/mL. The maximum excretion of sialic acid was
observed for
samples collected 4-8 hours postdose for all animals. The total increase of
sialic acid
excreted in the urine postdose was equivalent to 1.08-3.20% of the doses of SA
contained in
TA-2.
[00255] The bioavailability of TA-2 was estimated to be 2.53% and 3.73% for
Animal Nos. 102 and 103, respectively, based on the individual urinary percent
excretion
value following IV and oral administration. The bioavailability is 97.4% for
Animal No. 101,
which was markedly higher than the other two animals due to its low percent
excretion value
of IV doses.
TA-3
[00256] The background sialic acid levels were below zero for predose samples
of all three
animals.
[00257] Following oral administration of TA-3 in prepared tablets at 3250 mg
of SA per
Tmax was observed from 2.00 to 4.00 hours postdose with the peak
concentrations
ranging from 6.52 to 17.0p.g/rnL. The concentrations of sialic acid generally
decreased after
Tmax to levels below zero at 24 hours postdose for all three animals. However,
the half-life
could not be estimated for the three animals as the measurable data were not
enough to
88
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
identify the termination elimination phase or the extrapolation of the ALTC to
infinity
represented more than 20% of the total area.
[00258] The oral bioavailability of TA-3 was estimated to be ranging from
1.46% to
4.14%, based on the individual AUC(0-t) value following IV and oral
administration.
[00259] Concentrations of sialic acid of all the predose urine samples were
noted to be
slightly above LLOQ, ranging from 10.1 to 11.21,tg/mL. The maximum excretion
of sialic
acid was observed for samples collected 4-8 hours postdose for Animal Nos. 102
and 103,
and 8-12 hours postdose for Animal No. 101, respectively. The total increase
of sialic acid
excreted in the urine postdose was equivalent to 0.94-2.99% of the doses of
S.A contained in
TA-3.
[00260] The bioavailabil.ity of TA.-3 was estimated to be 3.49% and 1.51% for
Animal Nos. 102 and 103, respectively, based on the individual urinary percent
excretion
value following IV and oral administration. The bioavailability is 85.0% for
Animal No. 101,
which was markedly higher than the other two animals due to its low percent
excretion value
of IV doses.
TA-4
[00261.] In animals treated with TA-4, no siali.c acid was rneasureable beyond
30 minutes
postdose.
[00262] Following oral administration of T.A-4 in prepared tablets at 5000 mg
of SA. per
animal, most of the concentrations of sialic acid were below LLOQ with the
exception of one
dog (Animal No. 201), where the concentrations were slightly above LLOQ at 4
and 6 hours
postdose. Tmax was observed from 2.00 to 4.00 hours postdose with the peak
concentrations
ranging from 8.97 to 15.7 gg/rnL. The concentrations of sialic acid generally
decreased after
T. to levels below zero at 24 hours postdose for all dosed animals. The half-
life could not
be estimated as the measurable data were not enough to identify the
termination elimination
phase.
[00263] The oral bioavailability of TA-4 was estimated to be ranging from
1.57% to
2.09%, based on the individual AUC(0-t) value following IV and oral
administration.
[00264] All the predose urine samples had a measureable concentration of
sialic acid
ranging from 6.4 to 42.6 pg/mL. The maximum excretion of sialic acid was
observed for
samples collected 8-12 hours postdose for Animal Nos. 201 and 203, and 0-4
hours for
89
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
Animal No. 202 respectively. The total increase of sialic acid excreted in the
urine postdose
was equivalent to 0.54-193% of the doses of SA contained in TA-4.
[00265] The bioavailability of TA-4 was estimated to be L42% and 2.55% for
Animal Nos. 202 and 203 based on the individual urinary percent excretion
value following
IV and oral administration. The bioavailability of Animal No. 101 could not be
estimated as
the mass excreted of sialic acid postdose was set to be zero when corrected by
predose data.
TA-5
[00266] The background sialic acid levels of all animals were below zero for
both predose
samples and 15 minutes postdose.
[00267] Following oral administration of TA-5 in prepared tablets at 5000 mg
of SA per
animal, most of the concentrations of sialic acid were below LLOQ with the
exception of
Animal No. 201, of which were slightly above LLOQ at 4 and 6 hours postdose.
Tm was
observed from 4.00 to 6.00 hours postdose with the peak concentrations ranging
-from 7.79 to
15.3 lagirriL, The concentrations the sialic acid generally decreased to
levels below zero at 24
hours postdose for all three animals. However, the half-life could not be
estimated for the
three animals as the measurable data were not enough to identify the
termination elimination
phase.
100268] The oral bioavailability of TA-5 was estimated to be ranging from
1.47% to
1.96%, based on the individual _AUC(0-t) value following IV and oral
administration.
[00269] Concentrations of sialic acid of all the predose urine samples ranged
from 2.27 to
23.6 lig/triL. The maximum excretion of sialic acid was observed for samples
collected 4-8
hours postdose for Animal Nos. 202 and 203, and 0-4 hours postdose for Animal
No. 201,
respectively. The total increase of sialic acid excreted in the urine postdose
was equivalent to
0.02-1.70% of the doses of SA contained in TA-5.
[00270] The oral bioavailability of TA-5 was estimated to be 0.52 % and 2.24%,
for
Animal Nos. 202 and 203, respectively, based on individual urinary percent
excretion value
following IV and oral administration. The bioavailability of Animal No. 101
could not be
estimated as the mass excreted of sialic acid 'postdose was set to be zero
when corrected by
predose data.
TA-6
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
[00271] The serum levels of sialic acid were below zero for predose samples of
most
animals except for one dog, Animal No. 102, of which was slightly above zero
but below
20% of the LLOQ.
[00272] Following an intravenous dose of 25 mg/kg of TA-6, the concentrations
of sialic
acid decreased quickly to levels below LLOQ at the timepoi.nt of 4 hours
postdose, and then
to levels below zero at the timepoint of 8 hours postdose, for all six
animals. The
concentrations generally declined in animals, except that Animal No. 203
exhibited much
higher concentrations at 6 hours postdose when compared to the previous time
point. Sialic
acid was eliminated in dogs with the half lives ranging from. 0.56 to 1.40
hours.
[00273] The concentrations of sialic acid in the urine varied between each
animal. It was
noted that the sialic acid levels in urine collected postdose were lower than
predose for
Animal No. 201, with two of the three concentrations of postdose samples
detected as below
zero.
[00274] The IV dose resulted in 72.4-87.7% of the administered dose being
excreted in the
urine of five of the six animals. One dog (Animal No. 101) demonstrated
excretion of only
1.1% of the applied dose.
[00275] The individual urinary percent excretion of TA-6 was used to adjust
the data of
oral doses to account for the fraction of sialic acid absorbed (Fu, %). From
the data reported
herein, Animal No. 101 were observed to have much lower urinary excretion
postdose after
IV dose, which resulted in a markedly higher value of bi.oavailability
estimated for its oral
doses when compared with other animals in the same group.
[00276] In summary, after oral administration of TA-1 through TA.-5, low
sialic acid levels
were detected in serum, most of which fell below the limit of quantitation.
Peak
concentrations ranging from 6.52 to 40.8 1.1g/mL were observed from 0.5 to 6
hours postdose.
Sialic acid was eliminated with a half-life of 0.56 to 1.40 hours, calculated
based on the
serum concentration data from intravenous dose of TA-6. The bioavailability
was estimated
based on the individual AUC(0-t) value following IV and oral administration.
The
pharmacokinetic parameters estimated for sialic acid in serum are presented in
Table 21 to
Table 23.
Table 21
Pharmacokinetic Parameters estimated for simile acid in serum
R a ue
Test Dose Level Animal Cmax ti AUC(0-t)
Fs
Article (mg/kg) Number (pg/mL) h ) (jig
*biniL) (%)
9
CA 02862836 2014-07-02
WO 2013/109906
PCT/US2013/022167
TA-1 353-445 101-103 15.8-40.8 0.5-2 54.1-128 3.43-
6.76
346-508 201-203 12.6-20.5 1 43.5-66.7 2.73-
3.20
TA-2 361-451 101-103 7.98-13.7 2-4 26.5-62.4 1.64-
3.25
TA-3 353-439 101-103 6.52-17.0 2-4 23.1-77.6 1.46-
4.14
TA-4 538-746 201-203 8.97-15,7 1-4 38.3-74.4 1.57-
2.09
TA-5 543-781 201-203 7.79-15.3 4-6 35.7-73.6 1.47-
1.96
TA-6 25 101-103_ - 106-123 _.
.
25 201-203_ - 108-120 _.
.
- = not applicable.
92
Table 22
Pharmacokinetie Parameters of Sinlie Acid in Beagle Dog Serum Following Oral
Administration 0
tµ.)
o
,-,
Dose Level Animal Cmax Timm AUC(0-1)
MRT(04) T1f2 A UC(0-int) mw(0-inl) Fs c,.)
1-,
Test Article (mg/kg) Number (tigirial..) (b) (lug *hina.) --
-------- (h) (h) (lig *hind) (h) (4) =
o
o
o
o
TA-1 353 101. 15.8 2.00 54.1 2.63
a a a 3.43
406 102 26.3 2.00 68.4 2.45
a a a 3.44
445 103 40.8 0.50 128 2.07
1.41 137 2.45 6.76
TA-1 508 201 20.5 2.00 66.7 2.51
1.49 68.6 2.72 2.73
346 202 14.2 2.00 51.9 2.39
a a a 3.20
357 203 12.6 2.00 43.5 3.00
1.39 45.4 3.29 2.81
P
.
TA-2 361 101 7.98 4.00 26.5 3.51
a a a 1.64 "
.3
r.,
o 392 102 13,7 4.00 62.4
3.76 a a a 3.25 .3
451 103 111.9 2.00 50.0 3.19
1.28 52.3 3.48 2.60
.
,--µ
,
TA-3 353 101. 6.52 4.00 23.1 3.65
a a a L46 ...]
,
r.,
382 102 117.0 4.00 77.6 3.80
a a a 4.14
439 103 11.9 2.00 30.6 1.84
a a a 1.64
TA-4 746 201 15,7 4.00 74.4 4.41
a a a 2.07
538 202 9.69 2.00 52.6 191
b b b 2.09
562 203 8.97 2.00 38.3 3.37
a a a 1.57
IV
n
TA-5 781 201 15.3 6.00 73.6 4.59
a a a 1.96 1-3
543 202 8.20 6.00 46.4 4.70
a a a 1.83 cp
n.)
562 203 7.79 4.00 35.7 4.58
a a a 1.47
1-,
t..,
t..,
-4
a: parameter was not reportable due to the measurable data were not enough to
identify the termination elimination phase.
b: parameter was not reportable due to the extrapolation of the AUC to
infinity represented more than 20% of the total area. o
b.)
o
I-.
ca
-...
I-.
o
%,o
µ,0
Table 23 o
ct,
Pharmacoldnetic parameters of sialic acid in beagle dog serum following IV
administration
Dose Level Animal AUC(0-t) IVIRT(0-t) T1/2
AUC(0-inf) MRT(0-hit) CL WI
Test Article (mg/kg) Number (rug *h/mL) (h)
(h) (pm *h/niL) -- (h) (int./min/4) (L/kg)
TA-6 25 101 112 0.82 0.56 113
0.84 3.70 0.18
25 102 123 1.06 1.40 139
1.65 3.00 0.36
25 103 106 0.79 0.66 108
0.84 3.87 0.22
0
TA-6 25 201 120 1.08 0.86 121
1.13 3.44 0.26 0
K.
25 202 117 :1.27 0.99 119
1.135 3.51 0.30
K.
,...-.. 25 203 108 1.18 0.58 113
1.43 3.67 0.18
,..,
4-.
K.
0
F.
&
I
0
.4
I
0
FF
en
L-3
cil
b.)
o
I-.
ca
--.
o
b.)
b.)
I-.
cr.
-..1
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
1002771 All the predose urine samples had a measureable concentration of
sialic acid and this
was used to correct the total increase in urinary excretion of sialic acid at
12 hours postdose. In
contrast to serum, most of the concentrations detected in urine samples
exceeded the limit of
quantitation. The urinary excretion of sialic acid, as a percent of dose
administered was estimated
for each dosing occasion. Based on the assumption that the amount of drug
excreted in urine after
oral administration was a reflection of the dose absorbed, the bioavailability
of sialic acid was
determined based on the percent excretion value following IV and oral
administration. The urinary
excretion parameters estimated for sialic acid are presented in Table 24 to
Table 26.
Table 24
Urinary excretion parameters estimated for sialic acid
Range
Test Dose Level Animal Percent Excretion (0-I2hr) Fu
Article (mg/kg) Nunther %) %)
TA-1 353-445 101-103 0.43-3.56 2.47-39.1
346-508 201-203 0.93-2.57 1.29-2.17
TA-2 361-451 101-103 1.08-3.20 2.53-97.4
TA-3 353-439 101-103 0.94-2.99 1.51-85.0
TA-4 538-746 201-203 0.54-1.93 1.42-2.55
TA-5 543-781 201-203 0.02-1.70 0.52-2.24
TA-6 25 101-103 1.11-87.7
25 201-203 0-75.7
= not applicable.
Table 25
0
Urinary excretion of sialie acid in beagle dog following oral administration
k.)
o
,-,
Dose
Mass Excreted (0-12hr) Percent Excretion c,.)
1--,
Test Dose Level Animal Mass
Excreted OW -- (PO ----- (0-12br) Fu =
Article (mg/animal) (mg/kg) Number Predose 0-4hr 4-8hr 842hr
Corrected (%) (%)
o
cr
TA-1 3250 353 101 4652.55 149.630 13342.9 4314.71
17807.2 14085.2 0.43 39.1
406 102 2448.73 549.029 65305.8 481631
70671.5 68712.5 2.11 2.47
445 103 4206.50 313.356 101372 17403.2
119089 115724 3.56 4.06
TA-1 3250 508 201 2372.90 0 85288.8
0 85288.8 83390.5 2.57 rt./a
346 202 1177.24 21908.6 319.025 8998.13
31225.8 30284.0 0.93 1.29
357 203 1966.81 17.3023 53674.5 1286.44
54978.2 53404.8 1.64 2.17
P
TA-2 3250 361 101 4083.30 139.071 37257.4 942.062
38338.5 35071.9 1.08 97.4 .
392 102 4244.85 14977.6 76498.4 15826.0
107302 103906 3.20 3.73
00
451 103 4006.52 1003.46 73487.3 848.833
75339.6 72134.4 2.22 2.53
00
,,
cr
.
N)
TA-3 3250 353 101 5310.07 6980.87 482.029 27399.5
34862.4 30614.3 0.94 85.0 '
,--µ
..
'
382 102 4269.49 33345.4 64010.5 3337.18
100693 97277.5 2.99 3.49 .
..,
'
439 103 2936.63 1901.40 42627.6 788.657
45317.7 42968.3 1.32 1.51 .
r.,
TA-4 5000 746 201 2767.70 526.991 368.095 28316.5
29211.6 26997.4 0.54 n/a
538 202 5621.96 29695.1 709.442 25589.5
55994.0 51496.5 1.03 1.42
562 203 4074.90 385.991 73.4542 99426.6
99886.0 96626.1 1.93 2.55
TA-5 5000 781 201 1178.21 1269.32 372.873 63.4198
1705.61 763.046 0.02 rila
543 202 966.212 303.204 19168.7 281.709
19753.6 18980.7 0.38 0.52
562 203 2479.44 443.881 63164.8 23357.0
86965.7 84982.2 1.70 2.24 IV
n
,-i
nia - not applicable
cp
n.)
o
1-,
-1
n.)
n.)
1-,
cr
-4
Table 26
0
Urinary excretion of sialie acid in beagle dog following IV administration
Mass Excreted (ttg) Mass Excreted
(0-12hr) (ftg) Percent Excretion
Test Dose Dose Level Animal
(0-12hr)
Article (mg/animal) (mg/k0 Number Predose 0-4hr 4-8hr 8-12hr
Corrected (%)
TA-6 227.5 25 101 256.405 1941.56 509.021 274.902 2725.48
2520.36 1.11
202.5 25 102 3508.71 161656 10622.6 4086.45 176365
173558 85.7
182.5 25 103 6124.14 105.980 158924 5859.68 164890
159990 87.7
TA-6 162.5 25 201 3085.53 267.937 0 0 267.937
0 0
237.5 25 202 1977.36 165111 400.537 8095.96 173607
172026 72.4
227.5 25 203 1487.66 119734 50377.7 3394.26 173506
172316 75.7
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
[002781 In conclusion, the pharmacokinetics of sialic acid in different
formulations following a
single oral or intravenous dose in the beagle dog were estimated, based on the
serum and urine
concentration data in this study.
Example 6
Canine Pharmacokinetics Study Regarding Sialic Acid Extended-Release (SA-ER)
1002791 Overview: canine pharmacokinetics (PK) were obtained to assess the
absorption and
pharmacokinetics of SA-ER tablets at three different single doses in a
crossover study design in
normal canines. The cumulative dose effect of SA-ER was assessed when given in
three times per
day dosing for seven days.
(00280) Test articles:
- 325 mg SA-ER tablets (as exemplified in the Examples described above)
Gavage: Sial.ic Acid Powder dissolved in saline at 325 mg/mL (API)
[00281} Design:
- Four consecutive one dose studies were performed with 1 week washout in N=5
dogs; 5 tabs, 10
tabs and then 15 tabs
- One separate repeat-dose study was performed: 1625 mg/dose (=5 tablets) TID
every 8 hrs for 7
days (4875 mg/dog/day)
- Food fasted prior to dosing, fed ¨4 hrs post dose
- Bodyweights (BWs) weekly
- PK sampling on dosing days and urine collected (0-24 hr pooled/dog)
98
Table 27
0
w
N
Study design: single dose x 3 levels - crossover followed by 7 day dosing
o
,-,
(..4
I-,
0
0
0
0
0
.=N
imals rT eat- Dose iDosing Blood tor Serum Urine (store
Phase Week An
merit (per animal, PO) Regimen
(store at -91) ''`C) at -80 ''`C)
. . .
I . . .
.
1625 frig g5 x 325-
SA-ER mg t'ab' lets)
3253 mg (10 x 325- Snr$e dose., * Pre-study
2 5 rn, '.,a,e..s-
SA-ER 2. 6, 10 and
mg tablets) one day * Pre each
treakrent '
or
24 hr ( 2hr) P
4875 mg, fl5 x 325- (respective = I, 2, 4õ 8,
12, 8 24 ' 0
,3
N,
.., fema!,es SA-ER
rsn,st.:dr, p 00
mg tablets) Day 01 hr ryast-
dose .
N,
03
3250 mg SA
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[002821 Results:
Figures 9-12 show graphical data of the results
SA-ER attained reasonable steady state blood levels in canines
Serum levels showed a dose-dependent difference though not proportional to the
dose
level
Repeat dosing demonstrated some accumulation maintaining a significant trough
SA
level but no high peak
API gavage was better absorbed with a higher C. than SA-ER possibly due to
early
stomach based absorption
Goal was for broad, flat absorption curve, without a peak
Total sialic acid in the urine suggested a flattening of absorption at the top
two dose
levels
Overall, the canine PK data showed that on repeat dosing 3x per day, a trough
SA level
was maintained despite fast clearance at about 10x the background level of
free sialic
acid
Example 7
Evaluation of the Pharmacokinetics of Single and Repeat Doses of Sialic Acid
Extended-Release
(SA-ER) Tablets in Patients with Hereditary Inclusion Body Myopathy (HIBM)
(00283) This study was performed to determine the pharmacokinetics of Sialic
Acid
Extended-Release (SA-ER) after single and repeated dosing. More particularly,
the following
study evaluated the pharmacokinetic (PK) parameters of an SA-ER with single
doses at four (4)
dose levels in both a fasted and fed state, followed by 7-day repeat doses at
three (3) dose levels
in patients with hereditary inclusion body myopathy (HIBM).
Overall Design and Control Methods
1002841 This was an open-label, single-dose (in-patient) and repeat-dose (in-
patient and
outpatient) study of SA-ER in patients with HIBM. Orally administered extended-
release tablets
(as exemplifed in the Examples described above), each containing 325 mg of
sialic acid, were
studied. Repeat doses were administered on a three times a day (TID) schedule.
[00285] Each patient entered the study unit after all screening procedures
were completed
(Study Days -28 to -3) and study eligibility was confirmed for a 3-day single-
dose (fasted) period
(Study Days 0-3), followed by a 2-day outpatient wash-out period (Study Days 4-
5), readmission
for 2 days for a single-dose (fed) period (Study Days 6-8), a 5-day outpatient
repeat-dose
treatment period (Study Days 9-12), readmission for the final 2 days of repeat
dosing (Study
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Days 13-14), and discharge from the study unit the following day (Study Day
15). Each patient
received a follow-up telephone call approximately 1 week following final
discharge. Study days
may or may not have been on consecutive calendar days depending on enrollment
and dose level
staging.
Enrollment of Study Subjects and Assignment to Treatment Groups
[002861 Before undergoing any study-related screening procedures, each
potential subject
provided informed consent. Informed consent was documented by means of a
written, signed,
and dated informed consent form. The investigator determined the potential
subject's suitability
for the study by interviewing the potential subject and by performing per-
protocol screening
assessments. A sufficient number of potential subjects were screened to enroll
approximately 24
study subjects at approximately two (2) study sites. Patients who withdraw or
were removed
from the study after receiving test drug were replaced on a case-by-case
basis.
1002871 At cheek-in on Study Day 0, each patient who qualified for the study
was sequentially
assigned a unique patient number. This patient number identified the patient's
case report form
(CRF) data throughout the study.
Duration of the Study
1002881 Each patient could participate in the study for approximately 4-8
weeks, including a
14-day treatment phase requiring 7 overnight stays in the hospital unit or
Phase 1 unit.
1002891 Each patient entered the study unit after all screening procedures
were completed
(Study Days -28 to -3) and study eligibility was confirmed for a 3-day single-
dose (fasted) period
(Study Days 0-3), followed by a 2-day outpatient wash-out period (Study Days 4-
5), readmission
for 2 days for a single-dose (fed) period (Study Days 6-8), a 5-day outpatient
repeat-dose
treatment period (Study Days 9-12), readmission for the final 2 days of repeat
dosing (Study
Days 13-14), and discharge from the study unit the following day (Study Day
15). Each patient
received a follow-up telephone call approximately 1 week following final
discharge. Study days
may or may not have been on consecutive calendar days depending on enrollment
and dose level
staging. The estimated duration for an individual patient in this study was
approximately 4 to
8 weeks.
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Patient Selection and Restrictions
1002901 Inclusion Criteria
Individuals eligible to participate in this study must have met all of the
following criteria.
1. Must be 18 years to 70 years of age.
2. Willing and able to provide written, signed informed consent after the
nature of the study
has been explained, and prior to any research-related procedures.
3. Must have a documented diagnosis, confirmed by genetic testing, of
hereditary inclusion
body myopathy (HIBM), also known as distal myopathy, rimmed vacuoles (DMRV),
or Nonak.a
myopathy due to demonstrated mutations in gene encoding the GNE/MNK enzyme.
4. Willing and able to comply with all study procedures, including multiple
overnight stays
at a hospital unit or Phase I unit.
5. Sexually active subjects must be willing to use an acceptable method of
contraception
(i.e. double barrier method) while participating in the study and for 30 days
after receiving the
last dose of SA-ER.
6. Females of childbearing potential must have a negative pregnancy test at
screening and
be willing to have additional pregnancy tests during the study. Females
considered not of
childbearing potential include those who have been in menopause at least 2
years, or had
bilateral tubal ligation at least 1 year prior to screening, or who have had
total hysterectomy.
[00291} Exclusion Criteria
Individuals who met any of the following exclusion criteria were not eligible
to
participate in the study.
I. Pregnant or breastfeeding at screening or planning to become pregnant
(self or partner) at
any time during the study.
2. Use of any investigational product or investigational medical device
within 30 days prior
to screening, or requirement for any investigational agent prior to completion
of all scheduled
study assessments.
3. Ingestion of ManNAc, sialic acid, or related metabolites or sialic acid
donors that provide
this substrate in either chemical or nutritional supplement form during the 30
days prior to
screening. If ManNA.c or other substrate was used more than 30 days prior to
screening, the time
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period of use, the compound used, and the dose and dose regimen should be
recorded in the
patient's history. If a patient has been on substrate replacement therapy in
the past, the
investigator must consider the potential confounding effects of this therapy
before enrolling the
patient.
4. Presence of a condition the severity and acuity of which, in the opinion
of the
investigator, warrant immediate surgical intervention or other treatment.
5. Presence or history of any hypersensitivity to SA or its excipients
that, in the judgment of
the investigator, places the subject at increased risk for adverse effects.
6. Presence of a concurrent disease or condition that would interfere with
study participation
or affect safety such as swallowing difficulties.
7. Presence or history of any condition that, in the view of the
investigator, places the
subject at high risk of poor treatment compliance or of not completing the
study.
8. Serum transaminase (ALT, AST, CiGT) levels > 3 x upper limit of normal
(UN) or
serum creatinine > 2.0 mg/dL.
l002921 Prohibited Medications
Patients were not enrolled if they used any investigational product or
investigational
medical device within 30 days prior to screening, or if they required any
investigational agent
prior to completion of all scheduled study assessments. Ingestion of N-acetyl-
D-mannosamine
(ManNAc), sialic acid, or related metabolites or sialic acid donors that
provide this substrate in
either chemical or nutritional supplement form was prohibited during the 30
days prior to
screening and throughout the study. If ManNAc or other substrate was used more
than 30 days
prior to screening, the time period of use, the compound used, and the dose
and dose regimen
were recorded in the patient's history. If a patient had been on substrate
replacement therapy in
the past, the investigator considered the potential confounding effects of
this therapy before
enrolling the patient.
Patients were not permitted to use alcohol, tobacco or any nicotine-containing
product,
any caffeine-containing food or beverage, or grapefruit or any grapefruit-
containing product
from 4 days prior to screening to the time of discharge from the study unit on
Study Day 15.
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1002931 Permitted Medications
Other than the medications specifically prohibited, patients could receive
concomitant
medications as required. If a patient took any medication other than SA-ER,
the patient recorded
the date and time the medication was taken, the name of the medication, and
the reason the
medication was taken in the drug administration diary.
[002941 Any concomitant medications or other treatments were recorded in the
patient's
medical record and CRF along with the dosage and duration of treatment.
Clinical Trial Supplies and Administration
[002951 Formulation, Packaging, and Labeling
Sialic acid extended-release tablets (SA-ER tablets) used in this study were
white, oval,
film-coated tablets containing 325 mg of sialic acid active ingredient and
weighed approximately
780 mg (43% active) as exemplified in the Examples described above. The
tablets were for oral
dosing and were developed to have sustained release of the active ingredient,
SA, for up to 24
hours. All the excipients (inactive) contained in the tablet formulation met
USP or USP NF
compendia specifications and were generally regarded as safe (GRAS). No animal-
derived
products were used in the manufacture of the tablets. The drug product (the
tablet form) was
manufactured, packaged, and labeled according to Good Manufacturing Practice
(GMP)
regulations.
[00296} SA-ER 325 mg tablets were bottled and labeled. Each bottle was marked
with a label
that displayed the protocol number, the name and city, state, and zip code of
the sponsor, the
identity and strength of the contents ("Sialic Acid Extended Release Tablets,
325 mg"), the
number of tablets in the bottle, the lot number, the storage conditions, and
the statement,
"Caution: New Drug ¨ Limited by Federal (US) Law to Investigational Use."
1002971 Study Drug Administration
Patients received SA-ER tablets orally at one of four (4) dose levels in the
single-dose
phase and one of three (3) dose levels in the repeat-dose phase. During repeat
dosing, the total
daily dose was divided evenly into three doses given in the morning, in the
evening, and at
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bedtime (qHS) (see below). No placebo or active comparator was administered
and the study
drug was administered on an open-label basis.
1002981 Each of the 24 enrolled patients was sequentially assigned to a
specific dose level and
received two single-dose exposures at that same dose level (fasted and fed).
The low-dose
cohorts were filled before assigning higher-dose cohorts. The patient was then
assigned to
receive one repeat-dose regimen. The lower-dose repeat-dose cohorts were
filled before
proceeding to higher repeat-dose levels. Dose levels were as follows.
Single doses:
650 mg (n = 6)
1,950 mg (n = 6)
2,925 mg (n = 6)
4,875 mg (n 6)
Multiple dosing:
650 mg TID (1,950 mg/day; n = 8)
975 mg TID (2,925 mg/day; n 8)
1,625 mg TID (4,875 mg/day; n = 8)
[00299] Single doses of study drug were administered by site personnel while
patients were
confined to the hospital or Phase 1 units. For the repeat-dosing regimens,
each patient was
dispensed a 7-day supply of study drug at the sequentially assigned dose level
along with a drug
accountability diary.
[00300} Patients were instructed to take SA-ER with water three times a day
according to the
following schedule: morning (7:00 AM-9:00 AM), evening (5:00 PM-7:00 PM), and
at bedtime
(qHS; 10:00 PM-12:00 AM). In preparation for the possibility of forgetting a
dose, patients were
instructed to take a missed dose up to 6 hours after the appointed time but at
least 2 hours before
taking the subsequent dose. Patients were not to take a double dose. Patients
were asked to
swallow the administered tablets whole and not to crush or chew them.
[003011 As shown in the Study Scheme (Figure 13), dose levels were studied
sequentially,
progressing from lower to higher levels of exposure. At each single-dose
level, enrollment was
staged such that no more than two patients received the currently studied dose
level the first
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week for that dose level, with the remaining four patients for that dose level
treated the next
week or shortly thereafter, assuming no significant events occurred after the
first two patients
received the studied dose. Once all six patients had received single doses at
a given dose level,
then repeat dosing at that daily exposure (divided into TID doses) began.
[003021 As shown above, the maximum daily dose of SA-ER in this study was
4,875 mg,
administered as 1,625 mg TID. The maximum duration of administration of study
drug was a
single dose followed by a 2-day washout period and then a second single dose
(without
subsequent washout, if applicable) followed by a 7-day TID dosing period.
Study days may or
may not have been consecutive depending on enrollment and dose level staging.
[003031 Monitoring Compliance
Patients were confined to the hospital or Phase 1 unit during the single-dose
periods, at
which times study medication was dispensed and taken under the supervision of
site personnel.
For the repeat-dosing regimens, each patient was dispensed a 7-day supply of
study drug at the
sequentially assigned dose level and was asked to maintain a record of self-
administration of
study drug in a drug accountability diary. This diary was checked when
patients returned to the
study unit on Study Day 13 and collected by site personnel prior to discharge
on Study Day 15.
Site personnel maintained a record of all medication dispensed to each
patient.
[003041 Blinding Procedures
This was an open-label study. Patients were assigned to an open-label SA-ER
dose group
sequentially as they checked in at the hospital or Phase 1 unit on Study Day
0.
Study Procedures and Assessments
1003051 Types and Sequences of Procedures
The overall flow of the study is illustrated in Figure 13.
[003061 Screening Assessments
Potential study participants were screened approximately 3 to 5 days (up to 28
days
permitted) prior to admission to the study unit on Study Day 0. Screening
procedures included
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obtaining a medical history, performing a physical and neurological
examination, obtaining
samples for clinical laboratory tests (blood chemistry, hematology,
urinalysis, tests for
communicable viral diseases [hepatitis A, B, and C and human immunodeficiency
virus], and
urine pregnancy test [women only]), and recording vital signs (after 5 minutes
sitting, including
heart rate, blood pressure, respiratory rate, and temperature), height and
weight, and prior
medications.
100307] Following evaluation of the results of the screening assessments,
patients who met the
inclusion and exclusion criteria and who did not present any other reason for
exclusion were
considered eligible for enrollment into the study.
[003081 Screening Failures
If a potential subject provided signed informed consent but failed to meet an
inclusion or
exclusion criterion, or for any other reason was considered unsuitable for
participation in the
trial, he or she was considered a screen failure and was not asked or allowed
to participate further
in the study. The reason for exclusion of the potential subject and the date
and time the decision
was made to exclude the subject was recorded in the potential subject's CRF.
1003091 Single-Dose Phase (Day 0 to Day 7)
1003101 Admission and Administration of First Single Dose (Fasted)
Patients who met the screening criteria were admitted to the hospital unit or
Phase 1 unit
on the evening of Study Day 0. The medical history was updated and continued
compliance with
entrance criteria was checked. A physical and neurological examination was
performed,
including weight, and all screening clinical laboratory tests were repeated.
In addition, a blood
sample was obtained for total and free sialic acid analysis. Vital signs and
concomitant
medications were recorded.
[003111 Patients were sequentially assigned to a dose group on Study Day 0,
starting with the
lowest dose and following with sequential step assignments to higher doses.
Enrollment was
staged such that no more than 2 patients received the currently studied dose
level the first week
for that dose level, with the remaining 4 patients for that dose level treated
the next week or
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shortly thereafter, assuming no significant events occurred after the first 2
patients received the
studied dose.
[003121 Adverse effects (AEs) were monitored continuously throughout the study
(screening
through follow-up) by spontaneous reporting as events occur. Drug
accountability was recorded
following each dispensation of study drug.
[00313] On Study Day 1, blood samples were collected at the nominal timepoints
0, 4, 8, 12,
16, 20, and 24 hours to determine the baseline 24-hour, time-matched serum
levels of free SA
and to establish the diurnal cycle of SA levels. Actual blood collection times
were recorded and
were within 30 minutes of the scheduled collection time, except for the Hour
0 timepoint.
Patients received nothing by mouth, except water as desired, from 10:00 PM on
Day 1 (the night
before dosing) through 10:00 AM on Study Day 2 (day of dosing).
[00314] On Study Day 2, the initial single dose of study medication was
administered with
240 nil, of water at approximately 8:00 AM, and blood samples for PK
determinations were
collected within 30 minutes before administration and at the following nominal
timepoints: 10,
20, and 30 minutes and 1, 2, 4, 8, 12, 16, 20, and 24 hours after dosing.
Actual blood collection
times were recorded and were within 5 minutes for the 10, 20, 30 minute, and
1 hour
timepoints, 15 minutes for the 2 hour timepoint, and 30 minutes for all
remaining timepoints.
Vital signs (sitting) were recorded within 30 minutes predose and 6, 12, and
24 hours after
dosing. AEs and any concomitant medications were recorded.
[00315} On Study Day 3, samples were obtained for total and free sialic acid
at Hour 0 (the
same as Study Day 2 Hour 24). Vital signs, AEs, and any concomitant
medications were
recorded. Patients were discharged from the hospital unit or Phase 1 unit the
morning of Study
Day 3 after all Study Day 3 procedures had been performed.
[003161 Administration of Second Single Dose (Fed)
Patients returned to the hospital unit or Phase I unit the evening of Study
Day 6. At that
time, they received a physical/neurological examination, including weight, and
a urine pregnancy
test was performed, as applicable (women only). Vital signs (sitting),
concomitant medications,
and AEs were recorded.
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[003171 On Study Day 7, the second single dose of study medication (same dose
as Study
Day 2) was administered with 240 mL of water at approximately 8:00 AM, within
30 minutes
following consumption of a full fatty/protein meal. Blood samples for PK
determinations were
collected within 30 minutes before administration and at the following nominal
timepoints: 10,
20, and 30 minutes and 1, 2, 4, 8, 12, 16, 20, and 24 hours after dosing.
Actual blood collection
times were recorded and within 5 minutes for the 10, 20, 30 minute, and 1
hour timepoints,
15 minutes for the 2 hour timepoint, and 30 minutes for all remaining
timepoints. Vital signs
(sitting) were recorded within 30 minutes predose and 6, 12, and 24 hours
after dosing. AEs and
any concomitant medications were recorded.
[003181 Repeat-Dose Phase (Study Day 8 to Study Day 14)
On Study Day 8, samples were obtained for total and free sialic acid at Hour 0
(the same
as Study Day 7 Hour 24). Vital signs, AEs, and any concomitant medications
were recorded.
Patients were sequentially assigned to a repeat-dose regimen, as described
previously. Patients
were given an adequate supply of study medication for 7 days of TID dosing,
with full dosing
instructions including a drug administration diary or subjects were scheduled
at a later date to
return to the hospital or Phase 1 unit for a study medication dispensing
visit. Subjects took their
initial dose of repeat-dose study medication in the hospital or Phase 1 unit
prior to discharge.
1003191 Study days were or were not consecutive depending on enrollment and
dose level
staging. Therefore, most subjects were discharged from the hospital or Phase 1
unit on Study
Day 8 without receiving study medication for the 7 day repeat dosing. For
these patients an
additional study visit was required solely for the purposes of dispensing
study medication for the
7 days of TID dosing. This visit was scheduled prior to discharge from the
hospital or Phase 1
unit. This visit occurred 1 week up to 4 weeks following discharge from the
hospital or Phase 1
unit on Study Day 8. For the purpose of the study this visit was considered
resumption of Study
Day 8 for these subjects.
1003201 During this study medication dispensing visit, AEs and concomitant
medications
were recorded. Patients were given an adequate supply of study medication for
7 days of TID
dosing, with full dosing instructions including a drug administration diary.
After taking their
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initial dose of repeat-dose study medication in the hospital or Phase 1 unit,
patients were
discharged.
[003211 Patients returned to the hospital unit or Phase 1 unit the evening of
Study Day 13. At
that time, they received a physical/neurological examination, including
weight. Vital signs
(sitting), concomitant medications, and AEs were recorded. A count of
remaining tablets of
study drug was made. Patients continued to repeat TID dosing while in the
study unit.
[003221 On Study Day 14, blood samples for PK determinations were collected
within
30 minutes (Hour 0) before the morning dose of study medication (at
approximately 8:00 AM)
and at the nominal timepoints of 4, 8, 12, 16, 20, and 24 hours thereafter to
verify the steady-
state levels of free sialic acid. Actual blood collection times were recorded
and were within 30
minutes of the scheduled collection time, except for the Hour 0 timepoint.
Vital signs (sitting)
were recorded within 30 minutes predose and 6, 12, and 24 hours after the
morning dose. AEs
and any concomitant medications were recorded.
[00323] Discharge from Study (Study Day 15)
On Study Day 15, samples were obtained for total and free sialic acid at Hour
0 (the same
as Study Day 14 Hour 24). Vital signs, AEs, and any concomitant medications
were recorded.
Any remaining study medication was obtained from the patients, and no further
dose was taken.
Barring any residual safety concerns, patients were then discharged from the
hospital unit or
Phase 1 unit at the discretion of the investigator after all Study Day 15
procedures have been
performed.
1003241 Patients were considered to have completed the study after the final;
Study Day 15
discharge procedures were completed.
(00325) Vital Signs
Vital signs (recorded after 5 minutes sitting, including heart rate, blood
pressure,
respiratory rate, and temperature) were assessed at screening and daily during
the three in-patient
study periods (Study Days 0-3, 6-8, and 13-15). On Study Days 2, 7, and 14,
vital signs were
recorded within 30 minutes before dosing (before inital dose on Study Day 14)
and 6, 12, and 24
hours thereafter. Weight was measured at screening and on Study Days 0, 6, and
13.
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1003261 Concomitant Medication Monitoring
Use of concomitant medications was recorded when they were taken.
[003271 Pharmacokinetic Assessment
Evaluation of the pharmacokinetics of SA-ER included steady-state levels of
free, soluble
sialic acid in serum after repeated dosing.
[003281 On Study Day 1, blood samples were collected at the nominal timepoints
0, 4, 8, 12,
16, 20, and 24 hours to determine the baseline 24-hour, time-matched serum
levels of free sialic
acid (SA) and to establish the diurnal cycle of SA levels. Actual blood
collection times were
recorded and within 30 minutes of the scheduled collection time, except for
the Hour 0
timepoint.
1003291 On Study Days 2 and 7, blood was drawn for PK analysis within 30
minutes before
study drug administration and at the nominal timepoints 10, 20, and 30 minutes
and 1, 2, 4, 8, 12,
16, 20, and 24 hours thereafter. Actual blood collection times were recorded
and within 7.1-:
minutes for the 10, 20, and 30 minute and 1 hour timepoints, a: 15 minutes for
the 2 hour
timepoint, and 30 minutes for all remaining timepoints.
[003301 On Day 14, blood samples for PK determinations were collected within
30 minutes
(Hour 0) before the morning dose of study medication (at approximately 8:00AM)
and at the
nominal timepoints of 4, 8, 12, 16, 20, and 24 hours thereafter to verify the
steady-state levels of
SA. Actual blood collection times were recorded and within 30 minutes of
the actual
collection time, except for the Hour 0 timepoint.
[003311 Total Volume of Blood Collected
The total volume of blood collected for all scheduled study assessments was
approximately 219 rnL.
Statistical Considerations and Planned Analyses
1003321 Estimate of Sample Size
The sample size of 6 per group for the single-dose phase and 8 per group for
the repeat-
dosing phase was expected to provide sufficient information to meet the
objectives of the study.
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The sample size was estimated by evaluating the number of patients required to
assess PK
parameters. Based on prior historical PK studies in rare diseases, 6 to 8
patients per dose group
was determined to be sufficient for PK determinations, particularly with the
slow extended-
release profile expected. A sample size of 6 to 8 patients per dose group and
the total of 24
patients was also believed to be an indication of dose-drug level relationship
adequate for
planning dose levels for future studies.
[003331 Pharmacokinetic Analysis
Free sialic acid and its metabolites were quantified using a specific liquid
chromatography-tandem. mass spectroscopy (LC-MS/MS) analytical method.
Serum
concentrations of free sialic acid were measured, and the resulting data was
listed and tabulated.
Accountability of Clinical Trial Supplies
[003341 Storage and Handling
Clinical trial supplies were stored in a secure location at controlled room
temperature,
shielded from bright light, and kept off the floor.
[003351 At each pharmacokinetic sampling timepoint, 3.5 mL of blood was drawn
into serum
separator tubes and spun at 2000 RPM (or equivalent conversion) at 4 C for 20
minutes. Serum
was separated and transferred into two equal aliquots and stored upright in a -
70 C freezer prior
to analysis.
Results ¨ Pharmacokinetic Data
[003361 Figures 14-17 show PK data obtained for single dose fasted
administration for doses
of 650 mg (Figure 13), 1,950 mg (Figure 14), 2,925 mg (Figure 15), and 4,825
mg (Figure 16)
for human patients. Figure 17 shows PK data obtained for repeated dose
administration (650 mg
x 3; 1,950 mg) for human patients. Figure 18 shows PK data obtained for
different repeated dose
administration (975 x 3; 2,925 mg). The levels observed exceed normal patient
levels and were
close to levels observed in childhood before HIBM disease onset occurs. This
suggests that the
levels are clinically relevant. Further, the data of Figures 18 and 19
demonstrate achievement of
excellent steady state control levels without substantial peaks or troughs at
which the high
enough doses are twice that of normal continuously all day and all night.
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Example 8
Phase I Clinical ER/SA Study: Interim Safety Study
[003371 A Phase 1 study titled "A Phase 1 Study to Evaluate the Safety and
Phamiacokinetics
of Single and Repeat Doses of Sialic Acid-Extended Release (SA-ER) Tablets in
Patients with
Hereditary Inclusion Body Myopathy (HIBM)" was conducted. The study evaluated
the safety
and pharmacokinetics (PK) of single doses and 7 day repeat dosing of SA-ER in
HIBM patients.
The study was conducted in HIBM patients because the deficiency state of
sialic acid in HIBM
patients could fundamentally change the metabolism of the active ingredient
versus normal
healthy volunteers. The original protocol included doses up to 4875 mg/day
utilizing a 325 mg
tablet size (see Table 8, with Hypromellose), and was subsequently amended to
include an
additional cohort at 6000 mg/day utilizing a 500 mg tablet size. The 500 mg
tablet is the sam.e
formulation as the 325 mg tablet and was developed for the convenience of
patient administered
higher doses of SA.-ER..
[00338] The specific goals of this study were as follows:
Evaluate the safety of single doses of 650 mg, 1950 mg, 2925 mg, 4875 mg, and
6000 mg/day with and without food.
Evaluate the safety of repeat dosing of SA-ER at doses of 1950 mg/day, 2925
mg/day,
4875 mg/day and 6000 mg/day divided equally and administered three times per
day over 7 days.
Determine the PK. of SA-ER, including C. and AIJC of single doses with and
without
food, and steady state levels after repeated dosing. The background baseline
SA levels in HIBM
patients will also be determined.
Establish the best choice for doses to study in Phase 2.
Study Design
[00339] A total of 27 HIBM patients were enrolled at two (2) study sites but
only 26 received
drug. Subjects received SA-ER tablets orally at one of five (5) dose levels in
the single-dose
phase and one of four (4) dose levels in the repeat-dose phase. Enrolled
subjects were
sequentially assigned to a specific dose level and received two single-dose
exposures at the
assigned dose level (Fasted and Fed state). The subjects were then assigned to
receive one
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repeat-dose regimen. During repeat dosing, the total daily dose was divided
equally into three
doses, and given in the morning, in the evening, and at bedtime (qHS). No
placebo or active
comparator was administered and the study drug was administered on an open-
label basis.
1003401 Single doses of study drug were administered by site personnel while
subjects were
confined to the hospital or Phase 1 unit. Single doses levels were as follows:
650 mg (n = 6);
1950 mg (n = 6); 2925 mg (n = 6); 4875 mg (n = 4); and 6000 mg (n=6). For the
repeat-dosing
regimens, each subject was dispensed a 7-day supply of study drug at the
sequentially assigned
dose level along with a drug accountability diary. Multiple dose levels were
as follows: 650 mg
TID (1950 mg/day; n = 8); 975 mg TID (2925 mg/day; n = 8); 1625 mg TID (4875
mg/day; n =
6) and 2000 mg TID (6000 mg/day; n=6).
[003411 As shown in Figure 20, dose levels were sequential, progressing from
lower to higher
levels of exposure. At the 650 mg, 1950 mg, 2925 mg and 4875 mg single-dose
levels,
enrollment was staged such that at least two subjects received both single
doses (Fasted and Fed
state) prior to dosing of the remaining four subjects at the same dose level.
Proceeding with the
remaining four subjects was contingent on the safety profile observed. Once
all 6 subjects
received single doses at a given dose level, repeat dosing at that daily
exposure (divided into TID
doses) began. Subjects in the 6000 mg/day cohort were treated as enrolled and
not staged. The
lower-dose repeat-dose cohorts were filled before proceeding to higher repeat-
dose levels.
Study Results
1003421 Preliminary safety and pharmacokinetic results from the study are
presented below.
Summary of Preliminary Safety Results
Enrollment
1003431 Enrollment status and assignment to treatment groups of subjects are
shown in Table
28 and Table 29 below. A total of 37 patients were screened, and 27 were
enrolled. Twenty-six
(26) individual subjects completed dosing, one subject terminated early (prior
to dosing), and
eight (8) subjects withdrew consent before being dosed. All subjects dosed
with at least one
dose, completed all dosing.
[003441 In the 650 mg, 1950 mg 2925 mg and the 6000 mg single dose cohorts, 6
patients
were dosed per cohort. In the 1950 mg/day and 2925 mg/day multiple dose
cohorts, 8 patients
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were dosed per cohort. The 4875 mg cohort had 4 patients at the single dose
stage and 6 patients
in the repeat dose stage. There were 6 subjects total in the 6000 mg dose
level, both single and
repeat stages. Two subjects in the 6000 mg dose cohort had participated
earlier in the study in
previous dose groups (subjects 101-010B and 102-004B).
1003451 A total of 8 patients withdrew from the study before dosing and these
withdrawals
were due to the following reasons: 3 withdrew consent (personal reasons), 4
were screen failures
(I elevated GGT levels, I not a confirmed diagnosis of HIBM, 2 prohibited
concomitant
medication). One subject was an. early termination (Le., prior to the
initiation of treatment). One
subject was eligible and consented but was not dosed because the 6000 mg
cohort was full. No
subjects withdrew due to adverse events and no subjects withdrew after
initiating dosing.
Table 28 Patient Enrollment* Status & Dose Assignment
Dose Assignment
Subject ID Status
Single Repeat
101-001 Enrolled. 650 1950
101-002 'Withdrew consent.
101-003 Enrolled 1950 1950
101-004 Enrolled 2925 4875
101-005 Enrolled. 1950 2925
10:1 -006 Enrolled 650 1950
101-007 Enrolled. 650 1950
101-008 Enrolled 1950 1950
101-009 Withdrew consent.
101-010 Enrolled 2925 4875
101-010f3 Enrolled 6000 6000
101-011 Enrolled 2925 2925
10:1 -012 Enrolled 4875 4875
101-013 Enrolled. 2925 2925
101-014 Withdrew consent
101-015 Enrolled 487:5 4875
101-016 Enrolled 4875 4875
101-017 Enrolled. 4875 4875
101-018 Enrolled 6000 6000
10:1 -019 Screen Failure
101 -020 Enrolled 6000 6000
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Dose Assignment
Subject ID Status
Single Repeat
102-001 Enrolled 1950 2925
102-002 Enrolled 1950 2925
102-003 Enrolled 650 1950
102-003B Consented/Eligible Not
Dose(
102-004 Enrolled 650 1950
102-004B Enrolled 6000 6000
102-005 Enrolled 650 1950
102-006 Screen Failure
102-007 Early Termination
102-008 Enrolled 1950 2925
Table 29 Patient Enrollment*
Status & Dose Assignment (coned),
102-009 knrolled 2925 2925
102-010 Enrolled 2925 2925
102-011 Screen Failure
102-012 Screen Failure
102-013 Enrolled 6000 6000
102-014 Enrolled 6000 6000
*Enrolled = includes all patients who took at least one dose of drug.
B = denotes patients that participated twice in study at two different dose
assignments.
Table 29 Patient
Enrollment and Dose Group Assignment as Enrolled
Dose Assignment
Subject ID Status
Single Repeat
101-001 Enrolled 650 1950
101-006 Enrolled 650 1950
101-007 Enrolled 650 :1950
102-003 Enrolled 650 1950
102-004 Enrolled 650 1950
102-005 Enrolled 650 :1950
:101-003 Enrolled 1950 :1950
101-008 Enrolled 1950 1950
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Dose Assignment
Subject IT) Status
Single 1 Repeat,
101-005 Enrolled 1950 2925
102-001 Enrolled 1950 2925
102-002 Enrolled 1950 2925
102-008 Enrolled 1950 2925
101-011 Enrolled 2925
101-013 Enrolled 2925 2925
102-009 Enrolled 2925 2925
102-010 Enrolled 2925 2925
101-004 Enrolled 2925 475
101-010 Enrolled 292 =5 4875
101-012 Enrolled 4875 4875
101-015 Enrolled 4875 4875
101-016 Enrolled 4875 4875
101-017 Enrolled 4875 4875
101-018 Enrolled 6000 6000
Table 29 Patient Enrollment and Dose Group Assignment as Enrolled (Cont'd)
102-004B Enrolled 6000 6000
102-013 Enrolled 6000 6000
102-014 Enrolled 6000 6000
101-020 Enrolled 6000 6000
101-010B Enrolled 6000 6000
B=denotes patients that participated twice in study at two different dose
assignments.
Adverse Events (AEs)
Deaths and Other Significant or Serious Adverse Events (SAEsi
[003461 No deaths or significant or serious adverse events (SAEs) have been
reported in this
study.
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Adverse Events (AEs)
[00347} The summary of all AEs is shown in Table 30. A total of 31 adverse
events were
reported from 16 (61.5%) of the 26 subjects dosed. All events were rated as
either mild or
moderate. Three events were reported with an outcome of unknown: 1) bronchitis
that was
deemed unrelated to study drug; 2) cough that was also deemed unrelated to
study drug (these
two events were from the same subject 101-016); and 3) oedema peripheral
(finger swelling)
which began the day of the first 4875 mg single dose and was deemed possibly
related to study
drug (subject 101-017).
1003481 There were four adverse events rated as moderate: 1) a bruise from
a fall (subject
101-001) that was deemed unrelated to study drug; 2) fatigue/tiredness
(subject 101-008) which
began 3 days after a 1950 mg single dose, resolved within 1 day, and was
deemed possibly
related to study drug; 3) headache (subject 102-008) which began the day after
completing the
2925 mg repeat dose phase (i.e., Study Day 15 following discharge and noted
during the follow-
up phone call), resolved within 2 days, and was deemed possibly related to
study drug; and 4)
backache (subject 102-014) which was reported 2 days prior to initiation of
study drug, resolved
the same day, and was deemed unrelated to study drug.
[00349] Of the total 31 adverse events reported to date, 16 were possibly
related, two of the
16 events were moderate and resolved (as described previously), and the
remaining 14 events
were mild with all but one resolved. The one that was not resolved had an
outcome of unknown.
Of the 14 adverse events rated as mild, there were:
[00350} Five events of gastrointestinal (GI) disorders, three of which
occurred in one patient
(subject 101-003) at the 1950 mg dose and did not appear to be treatment
emergent, one event of
"heavy in stomach" that was mild and occurred on the first day of the 7 day
dosing period for the
1950 mg dose (subject 101-008), and one event of "dry mouth" that occurred
during the 7 day
treatment period while continuing therapy on 1950 mg (subject 102-005);
1003511 One event reported as mild fatigue at the 1950 mg dose that
resolved within one day
(subject 101-003);
1003521 One event of asthenia/all over body weakness that occurred on the
first day of the
2925 mg dose, was mild and resolved while continuing on therapy (subject 102-
002);
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[00353] One event each of back pain and leg pain in one subject (subject
101-013) occurring
on the same day and starting in the period between the single dose periods and
the 1 week
treatment period at the 2925 mg dose level;
[00354] One event of drowsiness on day 1 of the 7 day dosing period for the
1950 mg dose
(subject 102-004);
[00355] One event of sore throat on the second day of the 7 day therapy
period with the 1950
mg dose level (subject 101-008);
[00356] One event of mild headache in each of 2 subjects that resolved
within one day and
were reported on the first day of the 7 day dosing period at the 4875 mg dose
(subject 101-012)
and the 6000 mg dose (subject 102-004B);
[00357] One event of finger swelling (oedema peripheral) at the 4875 mg
single dose level
that was mild with an unknown outcome (subject 101-017).
1003581 The most common adverse event was GI disorders. Six total GI events
were
reported by 3 patients, and 4 of the 6 events were from a single patient
(subject 101-003) at the
1950 mg dose. Three of those four events in subject 101-003 were deemed
possibly related and
they occurred at the same time before and at the beginning of the 7 day
treatment period.
However, the timing did not appear to reflect a treatment emergent event and
there were no GI
events in the highest 6000 mg dose group. There was no pattern to the
gastrointestinal disorders
that suggested a treatment or dose related effect. The amount of sialic acid
being ingested is well
below the amount needed to generate osmotic diarrhea, i.e., loosening of
stools was observed in
the canine chronic toxicology study at the very highest 2,000 mg/kg dose
level, but this effect is
not relevant to the clinical study based on these results.
[00359] Among the general disorders, fatigue, asthenia or finger swelling
occurred in a total
of 4 subjects. The events lasted a day or two, and resolved either while
continuing in the study
or while on therapy (subject 102-002 at the 2925 mg dose). These subjects have
substantial
muscle weakness and fatigue is a common symptom of HIBM patients. There was no
pattern of
fatigue observed that would suggest a treatment emergent problem. There were
no events of
fatigue in the highest dose group 6000 mg.
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Table 30 Summary of All Adverse Events
n (/0 of number of subjects
Number of Subjects That Took Drug=26
that have taken drug)
Subjects that experienced any AE 16 (61.5)
Gastrointestinal disorders 3 (11.6)
Abdominal distension 1 (3.8)
Abdominal pain upper 1 (3.8)
Constipation 1 (3.8)
Diarrhea 1 (3.8)
Dry mouth 1 (3.8)
Flatulence 1 (3.8)
General disorders and administration site conditions 4 (15.4)
Asthenia 1 (3.8)
Fatigue 2 (7.7)
Edema peripheral 1 (3.8)
Infections and infestations 3 (11.5)
Bronchitis 1 (3.8)
Nasopharyngitis 1 (3.8)
Upper respiratog -tract infection 1 (3.8)
Injury, poisoning and procedural complications 4 (15.4)
Joint injury 1 (3.8)
Laceration 1 (3.8)
Limb injury 1 (3.8)
Procedural site reaction 1 (3.8)
Muscutoskeletal and connective tissue disorders 3 (11.5)
Back pain 3 (11.5)
Pain in extremity 1 (3,8)
Nervous system disorders 3 (11.5)
Headache 3 (11.5)
Somnotence 1 (3.8)
Respiratory, thoracic and mediastinal disorders 3 (11.5)
Cough 2 (7.7)
Oropharyngeal pain 1 (3.8)
Respiratory tract congestion 1 (3.8) --
Skin and subcutaneous tissue disorders 1 (3.8)
Rash 1 (3.8)
In summary, the adverse event profile observed to date has been unremarkable,
shows no dose
relationship and does not show a pattern of adverse effects that suggest there
is any reasonable
safety impact at any dose level. The range and type of adverse events observed
are common and
there was no dose dependent pattern for any drug related or unrelated event
that might suggest
SA-ER was having any discernible adverse effect in these 26 subjects.
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Clinical Assessments
Medical History
[00360} A comprehensive medical history was obtained at screening. This
history was
reviewed in the course of determining each potential sulzjectss eligibility
for enrollment.
[00361} In general, subjects demonstrated the expected profound muscle
atrophy and
weakness. A variety of other conditions were Observed that are typical for
patients in this age
group (29 to 61 years).
Physical Examination
[003621 Complete physical and neurological examinations were performed at
screening and
each time the patient checked in to the hospital or Phase 1 unit on Study Days
0, 6, and 13. The
neurological examination included assessments of cognition, cranial nerves,
motor function,
coordination and gait, reflexes, and sensory function.
[003631 There were no unexpected findings at baseline. Subjects generally
demonstrated
moderate to profound weakness in the lower extremities and upper extremities
that affected the
gait and muscle strength. There was no indication of increasing weakness or
loss of muscle
strength in this study.
Vital Signs
[00364] No significant findings were observed. In general, systolic and
diastolic blood
pressures were in the low normal range at baseline and during the study. All
other vital signs
were unremarkable. No significant abnormality was apparent and no changes
occurred with
treatment.
Clinical Laboratory Assessments
(00365) The clinical laboratory evaluations performed in this trial are listed
in Table 31.
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Table 31 List of Clinical Laboratory Tests Performed
Clinical chemistry Hematology
Alanine aminotransferase (ALT/SGPT) Hematocrit
Alkaline phosphatase Hemoglobin
Amylase MCH concentration (MCHC)
Aspartate aminotransferase (AST/SOOT) Mean corpuscular hemoglobin (MCH)
Bilirubin (direct and total) Mean corpuscular volume (MCV)
Blood urea nitrogen (BUN) Platelet count
Calcium Red blood cell (RBC) count
Chloride Reticulocyte count
Cholesterol (total) WBC differential
Creatine kinase (CK) Neutrophil count (absolute and %)
Creatinine Lymphocyte count (absolute and %)
Oamma-glutamyl transpeptidase (GOT) Monocyte count (absolute and %)
Glucose Eosinophil count (absolute and %)
Lactate dehydrogenase (LDH) Basophil count (absolute and %)
Lipase White blood cell (WBC) count
Phosphorus Urinalysis (routine)
Potassium Blood
Protein (albumin and total) Glucose
Sodium Ketones
Triglycerides Microscopic examination of the sediment
Uric acid Bacteria
Communicable viral diseases (screening) Casts
Hepatitis A surface antigen (HAV Ab total) Red blood cells
Hepatitis B surface antigen (FIBsAg) White blood cells
Hepatitis C antibody (HCAb) pH
Human immunodeficiency virus types I and 2 Protein
(HIV' and HIV2) Specific gravity
Urine pregnancy test (women only)
Abbreviations: SGOT, serum glutamic-oxaloacetic transaminase; SGPT, serum
glutamic-
pyruvic transaminase
[003661 A number of mild clinical lab abnormalities were observed most often
in screening,
but there were no abnormalities that were distinctly treatment emergent. The
following common
abnormalities were observed:
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[00367] In general, an elevation in creatine kinase was observed which is a
known finding in
HIBM patients and is expected. These abnormal levels ranged from mildly above
the normal
range to about 2-3 x the normal range and were present at baseline. There was
no pattern of
change with treatment.
[00368] Low creatinine was observed in the majority of patients. Creatinine
was usually about
Y2 the normal level at baseline and did not change with treatment. This
finding is mostly likely
due to the low muscle mass in HIBM patients which results in a decrease in the
total amount of
creatinine.
[00369] Low urinary uric acid levels were observed in these subjects at
screening, and this did
not change with treatment. The low urinary uric acid levels may be related to
low muscle mass
(like creatinine) and a low amount of adenosine-5'riphosphate (ATP) turnover
and purine
degradation. The no change with treatment suggests that SA-ER is not inducing
an adverse
secondary nucleotide metabolic turnover. That is, sialic acid requires
addition of cytidine
triphosphate (CTP) to become the CMP-sialic acid carrier. If CTP was being
excessively
consumed, this would lead to the need for increased ATP degradation to balance
nucleotide
concentrations, and would generate increased uric acid from purine
degradation. This
degradation pathway is not occurring based on these results.
[00370] Alkaline phosphatase was low in many subjects at baseline and this did
not change
with treatment. This result may reflect a lack of physical activity and less
bone turnover in
HIBM patients.
1003711 Lipase and LDH were marginally elevated in some subjects at baseline.
Also noted
were marginal low hemoglobins and/or hematocrits, variations in white cell
counts, modest or
borderline transaminase levels. There was no relationship to treatment
observed.
Safety Conclusions
[003721 In conclusion, the study drug was well tolerated in this group of HIBM
patients based
on the AE profile, the absence of SAE's, and the lack of treatment emergent
changes in any
parameter (AEs, physical examinations, vital signs and clinical laboratory
evaluations). The
adverse event profile observed to date has been unremarkable, shows no dose
relationship and
does not show a pattern of adverse effect that suggests there is any
significant safety impact.
Potential issues (such as diarrhea from the sialic acid load) were not
observed and there was no
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dose dependent relationship for the GI symptoms. Fatigue was observed in 2
patients but
resolved. Given the prevalence of fatigue and weakness in this population, the
lack of dose
dependent changes and resolution on therapy in 1 case, fatigue does not appear
to be a treatment
emergent phenomenon.
[003731 Based on the safety evaluation to date in this Phase I study, there
are no safety
concerns in proceeding to Phase 2.
Summary of Preliminary Pharmacokinetic Results
Enrollment
[003741 Data from 26 subjects has been obtained at 5 single dose levels
(fasted and Fed
state), and at 4 repeat dose levels for 7 days of dosing divided three times
per day (tidy. The
enrollment of patients in the different cohorts is shown in Table 32.
124
Table 32
Subject Enrollment*, Dose Assignment and PK Data Collection
1
Repeat Dose
C)
All Unique
Na
Status Subjects [a] 650 mg 1950 mq :-325
4675 r.,' 6000 mg 1950 mg 2925 mg 4875 mg 6000 mg
Enrolled
t..J
27
C.:
s4:
Have dosing data from:
s4:
Any time in the phase 26 6 6 6 4 ..
8 8 6 6
Day 2 26 6 6 6 4 6
.5
Day 7 26 6 6 6 4
Day 14 25
E, 8 5 6
Have Usable PK data from
Day:
1 26 6 6 6 4 6
,?. :3 6 6
2 26 6 6 6 4 6
7 25 6 6 4
14 25
!3 $ 4 6
Have Date Completed Cr
Discontinued
0
27 5 ;:. 1.4 ,.
E' E: 6 6
0
0
0
..,
0
0
VI
*Enrollment:
0
..,
,
27 = number of subjects who enrolled in study including one subject who
terminated early prior to dosing. 0
.1
I
26 = All subjects who took at least one dose of drug.
0
25 = number of subjects that have PK data available, sample tubes from one
subject were lost and there is a delay in reassaying back-up
samples.
v
r)
-3
Cl)
Na
Z
t. =J
t.a
t4
ii
ON
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PK Data Collection
[00375] The protocol put each subject within a cohort through a sequence of
free serum sialic
acid (SA) assessments at baseline, SA-ER administration, and collection of PK
and SA assessments
as follows:
[003761 Day 1: 24-hour monitoring of SA levels as a baseline to establish the
diurnal cycle of
free serum sialic acid levels.
[003771 Day 2: PK and 24-hour monitoring of S.A levels after administration of
the fasted dose
[00378] Day 7: PK and 24-hour monitoring of SA. levels after administration of
the fed dose
[003791 Day 14: 24-hour monitoring of SA levels during the 7th day of
administration of SA.-ER
in three times per day divided dosing. During the 7th day, SA-ER was
administered during
monitoring of the subjects to help establish steady state SA levels on chronic
administration.
Analysis of Free SA. levels After Single Doses of SA-ER (Fasted and Fed State)
[00380] Free serum sialic acid (SA) levels were determined by a validated
assay using a liquid
chromatography tandem. mass spectrometry (LC/MS/MS) methodology with a Lower
Limit of
Quantitation (LLOQ) at 0.05 micrograms/ml. The free SA data after a single
dose of SA-ER
(Fasted and Fed state) and at baseline are summarized for each cohort and are
graphically displayed
in Figure 21.
[00381] The mean baseline level of SA. was 0.143 (SD 0.0094) mcg/ral for all
enrolled HIBM-
affected subjects, which is significantly lower than the mean SA of 0.203 (SD
0.047) meg/mi for
normal individuals (samples obtained outside this protocol; n 47, age range 18-
78 years, and
tested using the same validated assay). Baseline SA levels do not reveal any
significant diurnal
variation in SA levels for any of the subjects individually (not shown) nor as
a group with fairly
tight standard deviations for the fiat baseline curves (Figure 21).
100382] The PK plots in Figure 21 show that the drug is being absorbed in a
generally dose
dependent fashion, with increasing doses leading to an increase in the size
and shape of the curve,
except for the 6000 mg dose (with 500 mg tablet) for which the curve had a
lower peak level than
the 4875 mg dose level (with 325 mg tablet). The PK. curves show increased SA
levels for 8---12
hours for most dose levels, with the 6000 mg dose achieving increased levels
for 12-16 hours. In
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general, the onset of absorption was earlier for the fasted treatment relative
to fed, and the fed
curves extended out longer than the more rapidly declining fasted treatment
curves.
[003831 The comparisons of the Fed curves (Figure 21) show that food had
variable effects in the
different dose groups. The PK curves were lower in the 650 and 4875 mg dose
groups with food,
similar in the 2925 mg dose group in both fed and fasted state and higher in
the 1950 and 6000 mg
dose groups with food. In general, the onset of absorption was earlier for the
fasted treatment
relative to fed, and the fed curves extended out longer than the more rapidly
declining fasted
treatment curves.
[003841 For most dose groups, the fasted curve hit a higher C., but for the
1950 mg and the
6000 mg dose, the apparent C.,õ, for fed and fasted is similar. The shift in
time and the broadening
of the exposure curve is particularly notable for the 6000 mg dose with the
500 mg tablet, which
shows a significant increase in blood levels for 16-20 hours.
[003851 When comparing the Fasted and Fed curves (Figure 22 and Figure 23), it
is clear that
below 4875 mg, there is a dose dependent rise in the SA levels (as expected)
and that uptake of SA
does not appear to be reaching saturation below 4875 mg/day. At the higher
dose levels, there is
increasing variability in the peak dose levels which represents very
significant inter-subject
variability in the absorption pattern for SA. In the 4875 mg single dose level
(n=4), two of the 4
subjects had very high levels of free SA (subject 101-016: 1.36 mcg/m1;
subject 101-015: 1.18
mcg/m1) as compared to normal (0.203 mcg/mL) and the two other subjects in the
same group
(subject 101-012: 0.79 mcg/ml; subject 101-017: 0.5 mcg/m1). The 6000 mg dose
group would be
predicted to have a little higher Cmax as compared to the 4875 mg dose (given
6000 mg is about 20%
more drug), but results were actually lower for the single dose PK. The reason
for this is not fully
established but may be due to the exceptionally high free SA levels achieved
in two out of four total
subjects in the 4875 mg single dose group (that may have randomly skewed the
data towards a
higher mean SA level in this smaller group), or a slower and more prolonged
absorption curve for
the 500 mg tablet as compared with the 325 mg tablet.
1003861 When comparing the PK plots for the "fed" state in Figure 23, the
difference between the
4875 mg dose and the 6000 mg dose is far less than in the fasted state. In
addition, the long period
of absorption for the 6000 dose with levels that are almost the same at 4, 8
and 12 hours, suggests a
more prolonged absorption curve relative to the other "fed" dose groups. The
other dose groups
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behave similarly with dose dependent absorption and about 12 hours of
significant exposure with
food.
Analysis of Free SA Levels After 7 Days of Repeat Dosing of SA-ER
[003871 Each subject was administered one of four repeat dose levels divided
into three times per
day dosing over a period of 7 days. On the 7th day of dosing (protocol Day
14), 24- hour monitoring
of SA levels was performed to assess whether steady and continuous exposure
levels were being
achieved. The PK curves over the 24-hour monitoring period are shown in Figure
24. Individual
repeat dose group PK curves are shown (Figures 24A-24D), and then all curves
are graphed on one
panel for comparison (Figure 24E). The last panel at the bottom right (Figure
24) shows a
comparison of mean SA levels over a 24-hour cycle for each dose group.
[003881 The curves (Figures 24A-24D) show that steady levels of SA are
achieved over the 24-
hour cycle with the trough never reaching the baseline mean SA concentration
of 0.143 mcglml. At
the lowest dose of 1950 mg divided three times a day (Figure 24A) (which is
650 mg three times per
day and dosed before morning meal, before evening meal, and at bedtime), the
curve reaches the
lowest point at 8 hours, but again achieves relatively steady levels over the
24 hour cycle. The peak
level is at 20 hours, and likely represents the confluence of the PM dose and
the bedtime dose in the
subjects. Given the physiology of muscle with nighttime anabolic activity, the
SA concentration
curve does show adequate coverage of the critical night period.
[00389] At the higher dose levels of 4875 mg or 6000 mg divided three times a
day, more
significant levels are achieved at all time points (Figure 24C and Figure
24D). Although the single
dose PK levels were very different for the 4875 and 6000 mg doses with
different tablet sizes (325
and 500 mg tablets, respectively), the two doses are comparable when given as
divided doses over
multiple days, and when we compare the 24-hour mean SA levels for 6 subjects
in each group
(Figure 24F). The longer PK exposure time of the 500 mg tablet may be
providing greater overlap
between administrations and thereby resulting in higher cumulative levels at
the 6000 mg dose.
When compared with untreated HIBM patient baselines, the two lower repeat dose
groups have free
SA levels about 2x baseline and the two highest dose levels are about 3x the
baseline SA level. All
doses resulted in free SA levels well above the normal free SA serum levels of
0.203 (SD 0.047)
mcg/ml.
I 28
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
Formal Analysis of Pharmacokinetic Parameters
[00390] PK analyses were performed using WinNonLin, version 5.3 (PharSight
Inc. Mountain
View CA, USA) to assess the standard PK parameters for the single dose data.
Given that there is an
endogenous level of SA, the proper calculation of PK parameters required the
subtraction of the
baseline SA levels in order to assess the changing levels for the administered
and absorbed SA.
Although there was no significant diurnal effect, the baseline SA. levels for
each subject were
subtracted from the corresponding PK level for that time point to create an
"adjusted" data point.
Therefore, whenever the term "adjusted" is used, it means that the PK.
parameter was calculated
after subtraction of the baseline SA levels. For timepoints in which there is
no corresponding
baseline level, such as the short 30 min time point from Day 2 PK, a mean
baseline level from that
subject was subtracted to determine the adjusted net increase in SA level over
baseline. These data
are shown fully in the PK tables in Table 33A to Table 33D, the key Cmax, AEC,
Tmax and T112
values are presented below.
129
Table 33.A: ;44s;14,;q nt PP
ftv.in ,ioxuao,:,::: ':,:,,1 ress 4-dales hoid
1
C)
AM: 0-24
b4
CD
iiub:lecn
OA= Inas lung x Tq R-squarva wa
".9 (al' 26nala iwyees .......... Mao*
V).4tr. Sccitflat4 (..ar) 44f4uki Oxr, ;0j ua
-,
10l-004 Snag Sisgit Post 9A-Sk. 1.94,U1 Pages.* Dose U.-4S. D44.41traci10
40zet Pgf :. 9.1S 9.99 .2.14 22:4.77 0.9199 wa
C:$
SxMgleiFaSting Dt4e z
0.32 1,09 1.2F, 9.20 0.5315 µ4,
9$nalef9ad Day 1
d.20 4.ft 4.10 v,
Repeat Day 14
d.3-3 t9.;ie 3.20 CD
ch
141-409 14tray Shollt Past 22-20. 1,64ag Atatss Pas* SA-S9 BaselItaino dws*
9mie 1 9,17 19.09 3.34 21.70 0.9921
Stalle;Tastiag 991F Z
O.
2.9S 0.64 T.91 0.92S9
SictlefTed t'we 1
9.99 4,99 1.44 4.S4 6.9999
Sweat Day 14
0.43 flo.:22 4.07
101,404 21,25aw dinwle nimuo 22.49 4424ag Naxsorm Dos* 22.-29 DasaldetAa Soso
Par 1 9.19 2.4.09 3..dd
Sizgle;Yasting Dar( 2
9.69 1.09 2.24 .*.i.39 0.9224
Sisyls:,s0 Pay 2
9.49 4.03
Repaat Day 14
0.62 12.i10 9.99 1.46 a.ernic
191-90 19ftaz V..Waigla Pea*9e-1P. 9999ut Sepast Poo* $2.14;! V44,41iVAINO
4**4 '04:;: 1
Sisylai7axtiaq Pay 2
9.3r.7 4.00 1.62 .=2õ24 9.9090
44.cylt.4a0 Pay 2
9.49 9.09 2.46 S..4'2 0.9099 0
lepeet Day la
0.46 4.40 9.29 $.61 i.dre42 o
m
m
X01-994 490at 94.441* tn.k* U-IP. .M0us 9eyeat Pa*t V4-XA 02**1.1nalflo 'lose.
Dei 0,16
to
wa fidayleinsaiag
Pay 2 0.4.2 2.09 1.42 Z..14 0.3,819 m
ua 41.agliai2t4
Pay 1 PAM 2.09 4,92 3.2,.11 0.2014 w
m
C:$
U14420; Pay 14
v.at; 16.09 1.44 MO 0.9499 to
a
I-
0* * aa-Y.P. 19VOug Myatt:. Pa** u-3vA romt*Ilum;m0 4,4aa 04t' :.
0.44 20,04 2.1,e A
1
Sir41efFe5tan4 Day 2
0.12 a.20 0.61 0
...1
I
Slaglt/244 Pay '?
2,14 2.02 4,20 a
Paws:. Pay 14
0.24 24.00 4,24 to
Wort: tii*oitd valet* ter ;,:ass, Iwo me A= 0-24 far subwas A01-010, 1.01,034,
arld 191-229 are 4st aa days. IM*M*Wit4W0441$. VtiAr adaslog IraAw3w
tor t 4 mai 9-Squared art da* ao lack at tda of she stgraato.m Ilat, at aka::
1 14 o*1424 not 8* c4.1m416t4d.
(a( 2eat xu.14ects 4nrol3e4 in tb* study rial,ot, Theo% sub9aers art
ideurlfiefiviahah* tirsa wurailaarc *aperient* I5.atit4 as a saa9ert so.AVSer,
ft:slab* staaladaspardeAeo liased v.:Tar:ha ZOWa 9.4494ww aron'oar and *0.
OA All peraueters at Bassalina/Ms daze ar* unadjustad (calculated ma the
COAAMOaXatiOU value at vlssxm.?.. 0s, all atlar days, tuax 2n9 Taax ass
unedu$tea;
492 1.-24, Tlel dad 279quared 4X4r ad3ustad (calculated wa east ak49441 from
bal.allat exteasharattaa. vatoa).
Eel 2,,:quared 12 24 lAdt224i0U iit ham mall the I.agtessica Iste tor the
teradral porwiae el the =kV* t1:415% the 4.1C1415V t.0 X.0, th* better: eht
lia,
while <0.S ir not a good fit. kulscupannaness *f tbm estinatod values have net
yet been evxduatod. Pagression results axe used *nly in tIta MO
t 15,41 Ii s'.
e)
tl
el
0*
b.)
CD
wa
ua
-,
em,
b.)
b.)
wa
ch
-...1
2.mtkre.15. Table 3313: . .g,R ft:SW.
P*4.SMA$A4U.A: f., Ism*: '25,F.12,4 ai..M4.
1
Dt.,t
'OgrAtiAt.t.134 0
Aor o=-21.
w
A.11"04C:t
eXerisk ftee cl'o.tW '2 T44: P-tqfmt4 c=
ID .el iSingla Repeat Phe5e ,Kisiu
0a-mgVILI1 '.>.".x bEfaL) 010 lel G..)
......õ
M,-Z,,U 1.914.itq .1.inli I0 Z,A-,.n 24.50pw ,s.t,,m&t. bi;tt, ..1.AAg 13o9-
mIt2N)5/14 8aa 0,a1 f',3 4,17 1.8.64 0,6261. 1-
e
Sirailefleaotfaai Day 2.
1.1..131 2.03: a67 32_12 6.66:31 0
Oi.w4le., :=ly 1
97 8,I2 1,99 113.130 V.t51
Par..aat Day 14
0.40 213.%; 3.16 0
..
Ch
.W.1.-Za0. 292...Sx1 -.in./U P4,:s4, $3.413, 4413.45.4...4.g13).$.:;.t.
4k...30. .PO.Uala=milk.,. 4Øit. t,k10 1. f1,.1,. 1.6,0' '..?....74
$'4:.(./6. 0.'.7t5'.
.
. .
/'tn11-tiVolmmic L,4sy Z.
1.1.4:'. 4.1> 1.e-; $"..51.- .11.4,01
Simialted nay 3.
101-n108 0088ag Omagla .f.fsee al-n 4o0t4a at Dora
9.&-4i Baza14ras.4a dura Day 1 0.17 1,:..08 3.64 48_04 0.2000
6t2,g1tf7g61sql Dzq Z
0.46 2,,8c, 2.14 1_44 3.9746
tir4laited Day 3
0.413 ok,08 4.41 3.0$ *.l'eeten
tweet Day /4
0,49. 20,OD 4,47
/01-011 2.97.47.,kg nx,..gle Duse 413.-.3.4 z824.8.1 tweet D*s.s, 02-Et
inszalimaina dura Day 1 3,21 14.'18 4,31 04,52 0,6363
:-e0.4.165,1mirmk-Aov .V4y Z.=
0,30 13.00 0,70 13-1.13 0-13131313
Singlaltled nay 3'
v,88 4,05: 1.21 1.72 J2,49ws
9Ar848
318s, 24 0.13," 0,00. ?.... 24,1$ .2113.13
P
.01-4)Ig 401;x1 9i8.11g: Iges,p0, U,D 48136130 D.4344. 44.4 $.a.-r.O.
Wm:at:milks clmt, 'Vq* 1 0,20 1.9, 4,1S 0
I.,
SiDqls;FartLin9 Day Z
3_79 3.52: 4.3/ 2.94 0.9557 .
ogA41,:dgeog r8v '.:e
v-we- :5,2.5 13..74 2..41 0.A.:71 Iv
m
t.,.) Repast Bay 14
0_91 4.06 10.26 w
m
Iv
.01-3.113 292.76ag 51rox2e Ca 5.1i,K4 2926224 Rapeat Dos* 3.1,-E13 Baseltua/Nu
dusa Day 1 11:23 20.0:2 4,11 0
r
5jX.gi4j344kXfs9 4.e4y 1
0,38 13,00 4,14 4-0 0A1344 A.
1
Sio4lanced Day 7
0_41 4.02 13.94 4.97 17!..sae4 0
134308.81:: 00-y /4
0.134 '24,00 4,013 ...3
1
0
I.,
Xote: Xis.sire values tar Cnar. 1511eX sca4 AUC 0-Z4 fzr 5nsjft.t.fts 101-020,
202-016, md 101-030 ara dun to data inconvirtanatea. Other alsalnq raImax
t*r T ka *ad 4.-VeasPt4 eee 4,19. ffe IS'AU 45= fat. o13 f3 smvsmtzl.m 12mt, m
tl.oM T ,* fmo.1.5I.:it bs. 4..*..1,
r.si.1 8ama rabtarts vaxalled irs, tiro rtudy tairs. Mara rUbflacts ara
idautitied rrith tha flaxt. enrol/mem exparianta 1iar,e3 as a s,2-act ismber,
mdtai& mcm4 molveimm 151384olth tk& zmo =0,t41.: 95x41347 echa 4 .2.
ikA A.11 parameters at Daaelineftto dole at raradjuatad 4calrraistad ua tha
cancentrazion value at visit$. On all other daya, Znat and laaa at*
=610=1:44.; Atte :',.:-.ZiA,. Tail Afta .3-Irms.k.1, ts.s.*. sa.tivoet44
cAmimaz13m4 m tam Mpsmos tx4m Mamal.sul :f*F2A.fOltt4tft..0e V4.11).41,
E03 2-snuarad is ar.,indlat.:ion ef hew weIl the regress.leu lice 2or the
terainsi poztAus: of tha cuxua 'fits: the clw.say. to 1.0, the h.c.4:tex the
it,
34414 'q3, 246 mt, .z. got.M. ti'v. Apprmamgomm ot the .4..=MAz*dw.1ut hav.*
wt. ye,t, ..t..mn 53'4l)),51344, PAImmim. smmatz :4.tt m45.61 0i17 &o. Mo
estamatiun u half Lite.
00
Data 51. 4
n
cp
w
..
w
,
w
w
Ch
-...1
Table.33B: Liaain af DK Darlaa
Ni.x8a4nwra fag -fgaa fta.11.2 .a.old 1
C)
A.IX *.-e4
0
SsXt..ject
ais&X TfliOX so.og. x Pe S-squered
(....)
31' Eal Sin:;1.4: S...4lat:
PheZe ViZIn Oa!: ,,en3-..? inal
1-,
4P4-04$ 491144 9,1alla Paaw '22.--Bt 4P7Pay Dwazat D*aa 94.-g9. Pazw4inanao
**.4 Pay .4 6-2'2 14.P0 Z.0 29.44 ,..:1.6674: 0
BingIVYastiw
Day 2 1_1'8 Z..:.1e3 3.7S 2,75
Vinala;red
Day 7 ".5.42. 4,07 2:.Z6 4.,./1.S 0.79Da: 0
9apaat
Oav 14 D...4V. t0,00 4.We, cT
161-014 41:1'1",ay Dinala Paza 22.-8t 4a78- Aapzat 0asa 92.-X9. PasaS:ina,14*
4aa* Day 1 6,IS 14.'64 3.%.::1 27.14 6,064.4
dinwla?fazwaa
2.,24, 2 4 36 1.27 7.43 1.6a 6.6064
dixar2aelrad
Day 7 6..67 8.J.10 4.94 3.61 1,0360
hapaaO
Boy 14
1B2-017 4e7Sag Single B.
SA.-IS 487Seg. Repeat Roes SA-IR Barieldnellio doze Day 1 3.14 35. BO
3. ;LS 48.83 6.9969
Dimiacicaraa4
Bay 2 6.60 4.17 4.M S.S: t''.: DADO
Bingl:e?Yed
Day .3. 8.47 8. %30 4.M 2..84 D . V33
R61:002At
PO? i..1 ;1.71 1F..:.1,7 9.22 10.t11,3
lot-ola 600Smg Sim/lei 0naa.Ka..,,Ilf: 60D0x0; Repean Masa DA,22.
Daaalinalptaxe Zay 1 5x..2a 0.-0(1 3.87 E3,63 B..mEs
-stiti;41.4
t;.Ø 2 881 4.88 8.43. .87 (5.4841
faz4140.44
Day 7 D.49
$33 4.M 3.1..$ C,991;$: P
aavaam
Day 14 ;Las a.;:o 11.02 547_38 a_o887 m
r.,
m
m
DU-na 90D8aaj Z4nala Paaw u.--7..t. 1(3N51; p.vom be
ZA,29, Paaaw.lizaiRa 9aaa Pay 4 8.38 7.9.2 f.:91 .7.38.23
P.D4a0
1-,
0
(....) O'ingiefFesting
Bey 2 0.E8 2.00 2.2? 1..e? f3.9924 w
a
N Vinaloc9a4
Do/ , I.,
Dapaat
Day 14 8.49 24,00 4,85. 0
r
A.
1
=2-061 1.9.Wa/ 9,=,n.yla Paaw .,2. 81 Mana Uazza b*sa 0A.-49. 461mi-tan/A,
e,aaak Pay 4 6,I3 MO
diza440Zaatana
Pay Z 6,30 :33 28 e,*7 cocs 2.3774: ...3
1
ging-la/Fad
Day 7 ..a.1 4.03 1,26 1,64 D.933õ.2 0
I.,
Kespaan
Bay 14 D.29 20,00
gete: Missang vaioes tor Caza, Mmea ..%d AUC 2-24 for sclojezte lei-Lux, 101-
816, asz..1 101-023 are :due ta data incoasiswalzies. Other laising values
'Kat. 2 Z. ana A-taxed axa .D4r4 a4 lack al fit oft-ha gaga:oast:at lina, a*
that. T ki aun1dnam a* nab:ft:slats:6,
M ::::Wo$ aqOacta. matollad in tha atady taica. The9.4 intoaatz zaa idantiliad
wah tha kitza atxt11azza avaIdanaa 3izna6 az z. z...0ww nasapas:,
widths second empariance listed with the same rah:jam nmahax and a B.
RO All pagaaatago az Dalel3naltle, doat 824 mo.6loo.24d .aY4i.8a7ar4d on nha
C=gentr4ZiOXI:V41U4 42 VIelt.. a.). atl azhar ..daya, Max and. TWG axa
una4laatati.i, .k176 6-24, 41/2 aad B-aquagad We* adlazto4 4,:a1calated aa nha
chaaga txon azaalina aaaaanngandon walt5a.
a-squared iz en ixdScation af how well the xegres..rsion line 29T the termixel
porti:om of thA cum-ve tinsz nhe olt.-6er to 1.;.%, the hatter the tit,
aNiAa .40.1 ta: nea. A 8A4 *tie. Apprwii&t.~: 441 'nhe Qe,t42WWW3,Veilli.
..;AAV* nem T..Rt b.t.R.p.. wisaqb.t*A, Im,11.ftri. -:.:-.RwaITA wo, zaad way
:es tha 00
aztiaaaiaa 0 Pali 3444.
r)
,-q
cp
k4
o
,-,
L..)
.....õ
o
k4
k4
,-,
o
-4
Appendit Table 33C: PK Ser,,m. Pareeeters
for Free Sialic Acid
CD
---------------------------------- Dose -------------------------------------
------ Parameters(h) ------
ABC 0-24
ket
0
albject Oasx
Imax (lacg x 11-2 P.-squared
ID Da] Single Repeat Phase Visit
:10.cg./al.j 1]:ci hr/m1) Oir:, [c] CA)
......_
102-002 1950aci Simile Dose SA-ED. 2925mu Repeat Dose SA-ED. BaselinetWo dose
Day 1 0_1E 20_00 3_27
0
Single/Fasting Day 2
0_47 4_00 2.55 2_86 0_9890 0
Single/Fed Day 7
0.29 8.00 1.63 4.63 0.7248 0
0
Repeat Day. 14
0.38 20.10 3.45 0
102-003 550g Single Dose SA-ER 1950mg Repeat Dose SA-ER Baseline/No dose Day 1
0_16 16.00 3_57 211.64 0.7160
Single/Fasting Day 2
0_20 4.00 0_41 8_40 0_9986
Single/Fed Day 7
0.19 8.00 0.22
Repeat Day le
0.31 20.00 1.77
102-004 550mg Single Dose SA-ER 1950mg Repeat Dose SA-ER Baselineffeo dose Day
1 0_15 23.70 3_24
Singie/Fasting Day 2
0_23 4.00 0_87 3.59 0.8356
Single/Fed Day 7
0_19 8.00 0_60 1_91 0_8866
Repeat Day 14
0.33 20_00 2.93
102-0045 6000mg Single Dose SA-ER 6000mg Repeat Dose SA-ER Baseline/No dose
Day 1 0.16 16.32 3.39 66.84 0.8003
Single/Fasting Day 2
0_71 8.00 5.26 2..70 0.9937
Single/Fed Des/ 7
0_52 12.00 4_66 3.93 0.9995 P
Repeat Day 14
0.74 20.00 7.38 0
NO
a
102-005 6.50mg Single Dose SA-21). 1950mg Repeat Dose SA-ER Baseline/No dose
Day 1 0.17 20.09 3.59 m
re
Single/Fasting Day 2
0.39 20.00 1.47 0,
Goe Single/Fed Day 7
0_21 4.00 0_55 4.03 1.0000 w
ON
Goe
Repeat Day 14
0_27 16.00 1_49 13.53 0.8434 N,
0
r
ae
102-008 1950mg Single Dose SA-ER 2925mg Repeat Dose SA-ER Baseline/No dose Day
1 0.16 23.92 3.32
O
Single/Fasting Day 2
0.21 2_00 0.78 3.39 0.6418 ...3
1
Single/Fed Day 7
0.22 24.00 0.97 9.50 0.2.531 0
Repeat Dal, 14
0.28 24.00 2.37 72.21 0.1123 No
Mote: Missing values for Omar, Teas and AOC 0-24 for subjects 101-010, 101-
016, and 101-020 are due to data inconsistencies. Other missing values
for T li and P,Sguared are due to lack of fit of the regression line, so that
T 44 could not be calculated.
[a] Same subjects enrolled in the study twice. These subjects are identified
with the first enrollment experience listed as a. subject number,
and the second experience listed with the same subject number and a E.
(h] All parameters at Baseline/Mb dose are unadjusted calculated an the
concentration value at visit).. On all other days,. Ouax and Tax are
unadjusted; AMC 0-24, 31/2 and A-squared are adjusted .calculated on the
change from baseline concentration value.
[c] R-sguared is an indication of how well the regression line for the
terminal portion of the curve fits: the closer to 1.0, the better the fit,
while 40.5 is not a good fit. Appropriateness of the estimated values have not
yet been. evaluated. Regression results are used only in the 00
estimation of half life.
r)
Data as
1.q
cr
ket
0
Goe
....õ
0
ket
ket
0
-...1
Appeadia Table 331): t PK %tam Petamem0 tar Vrae ftelit Acid
CD
N
am ii-;1."4
o
SAArject
r.max Tmax (z.o...4 :a T.i. R-7.471.8.1-11,3.
I,
(44
p W Viwle Re7'.at Matt Viait
0ac,.W.,.? Ma:) ar.fal0 Mt) :r.f.11 --
1-,
10Z-009 MU4
9as9.1a 04Y4 $a-au aZiaq gevaat 9as,a ea-30; OamUna,lia dato Da," 1 0.11
0.00 ,40 194,1Z 0,037 0
Single/FaMnw Dav. 2
0.6,7 3.27 4.25 2.73 0_9/10
na014iPel. 1,,ey 7
0,49 4.M 2.91. 4.0 0.7607 0
P.eptat 9a3' 14
0.41 mm 4,99 cT
102-01.0 1925esl- 215)41a DaSe 2A-K.P. 02.1411 bakaat 9naa .1.1a-K9
Zaaalluailla date Day 1 0.14 4,00 ::'.1.1.4 19e,91 0.220
itiantalFaat.mg Dare
1 0.91 4,00 /.99 3.27. 0.7049
Single/rad Day 7
0.40 4.00 Z.49 4.7Z 093z
Peptat reaq 14
0..44 20.00 4.2.4
1$32-ol3 6900ag Singla Dee:e SA-IR 6000ag :Repeat. Doze SA-EP Baseline/Hu the
Dey 1 0.17 0.00 S..74
nalla/Yaaring Dgf I
0i:4 2 Or; 1..40 4.3. 0.2127
Simele/Fed Day 7
0.36 12.00 .T....14 2.34 0.9220
R4p4at D45. 14
0.:30 2Ø12 2.61
101-014. 20E1Ong gingle.D6M..gA,22, Zealiaq Repeat' 9080 0Axg0 0ezelli3e-
33t.A,y4a Baq, 4 Ø'.17 ZEI,041. 3,72
2ial/ai9satin9 Day 2
=0.4I 0.110 2.80 .2..1 0.9721
nn1.14/44 Da? 7
0.42 12,00 3. 94.42 1.0000 P
Repeat
Da y 14
0.20 20.00 9.51 0
Iv
0
Iv
I-,
0
(44
w
4=,
Iv
0
r
a.
1
0
...3
1
0
Iv
Nate: Missins.4 .u.aluz for Omen. Tasz and AUK: 0-24 tor sUizeet.s 1d1-010,
/0/-012, and 101-= are the t.:o. (1.,. inz.onsisteacies. a:her sissirg values
ker T and. a-9quayad aya dva t.e 1aria et tit 0t tan ragyaaaim Ilyse, es's
that T qu*uld nat. be reIt0lezed,
W 9caa aubf.';acm enzol1ed. :h. IOW f/ZUO MriA*. Mem atavj:eme fa*
idaAvsks!lel'ssith -the tirat ear*lhaarc a:spay:tame 1ietwa 0 a aWlaat.
meter,.
end the eacand experience listed vtza the lame au17:ect. sanher and a B.
04 Ail peraratara 41r 0atu1inait4o Mat wess mr,A4:11.utA4 ix:41c34-t-sn4 m th,
cennanaratian value at. ,gieW 0a all attaf: dart, 0itx ona Inav. ara
uneekJuated.;, A= 0-04. TI..OZ za4 9-amaze1 *24, 95415aata4 (aalculamd to the
chama ..on taaallne ..iameaMeatiam Valeal.
.:,:'.g 33-anus/Ed is am in/teat:Len of iSOTS TerZ.S1 the regre.6.sion line
for -,-..he tezzlna.3. porti.to of- the vArce tits: the closer. to I. 0, the:
better the fit,.
Vacita ,..D.I ia aOr: xi 5:1,y,4 tit. Apyffefvtiananeaa :it the tetie.aaaa
valaai biaa m..n. yet 3.;g01, MWOhehest PA9eale:tais taMltaa '4M,l q4a4 anW in
nhe 00
4at.$.5400..cal ot SI,s112 ¶u...
r)
,-q
Data OW S
cr
N
0
I-,
CA)
,
0
k.)
N
I-,
01
--11
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
[00391] The Cmax data (Table 34) shows that in general, fasted data. (Day 2)
showed the highest
C., for all dose levels except for the 1950 mg dose level. The highest Cmax
(0.958 meg/m1) was
achieved in the 4875 mg dose group (n=4), using the 325 mg tablets, but the
range of values was
about 3 fold (i.e., from 0.423 to 1.36 mcg/m1) within this dose group, The
Cõõõ achieved at the
6000 mg dose level (n=6), using the 500 mg tablet, was lower at 0.520 mcg/ml
but with a much
tighter standard deviation and range of values from 0.335 to 0.711 mcg/ml,
Interestingly, during the
repeat dose phase the two top doses of 4875 mg (ri=6) and 6000 mg (n=6) had
similar Cniaõ levels of
0.676 and 0,661, respectively.
135
Table 34 Cmax (mcg/m1) Values Following Single Dose
Administration (Fed and Fasted) and Repeat Doses of SA-ER
1
Parameter Phase Visit Statistic 650 mg 19!,e
x..j 2925 ma 4675 mg 6000 mg C)
Number of Subjects Single N 6 6
6 4 6 Na
Who Took Drug Repeat N 8
8 6 6 ....
t=J
Coax (mcgimL) Baseline (No Dose) Day 1 N 6
6 6 4 6
Mean 0.157 0.160
0.174 0.151 0.162 ...T.:
4.^.
SD 0.009 0.022
0.043 0.031 0.017 4.^.
Median 0.156 0.157
0.178 0.143 0.167
Min, Max 0.146, 0.172
0.134, 0.198 0.110, 0.226 0.122, 0.195 0.131, 0.179
CV(%) 5.919 13.755
24.783 20.776 10.225
SingleiFascing Day 2 N 6 6
6 4 6
Mean 0.296 0.315
0.600 0.958 0.520
SD 0.110 0.093
0.282 0.396 0.134
Median 0.274 0.305
0.538 0.984 0.507
Min, Max 0.183, 0.449
0.206, 0.470 0.300, 0.985 0.503, 1.360 0.335, 0.711
CV(%) 37.068 29.353
47.107 40.295 25.676
Single/Fed Day 7 N 6 6
5 4 5
Mean 0.187 0.326
0.434 0.629 0.494
SD 0.025 0.106
0.051 0.240 0.124 0
Median 0.192 0.326
0.413 0.571 0.489 o
Min, Max 0.139, 0.209
0.213, 0.485 0.379, 0.493 0.423, 0.950 0.356, 0.685
m
m
CV(%) 13.277 32.627
11.834 38.111 25.183 m
m
ma
m
Gra
w
m
CA Baseline (No (:'ose) Day 1 N
8 8 6 6
m
Mean 0.163
0.160 0.160 0.162 o
r
SD 0.017
0.039 0.028 0.017 m
1
Median 0.160
0.150 0.161 0.167 o
Q
Min, Max 0.146,
0.198 0.110, 0.226 0.122, 0.195 0.131, 0.179 1
o
CV(%) 10.105
24.447 17.602 10.225 m
Repeat Day 14 N 8
8 4 6
Mean 0.333
0.393 0.676 0.661
SD 0.095
0.093 0.180 0.282
Median 0.27
0.395 0.635 0.618
Min, Max 0.239,
0.531 0.280, 0.555 0.520, 0.914 0.346, 1.050
CV(%) 28.45
23.806 26.575 42.624
v
n
NOTE: Baseline values for single and repeat dose levels slightly vary because
some patients who are included in one dose group for the single
dose phase are included in a different dose group for the repeat dose phase.
Therefore, the baseline means attempt to reflect the mean of the cn
Na
same patients that are being compared, and the mean changes slightly when you
take some patients in or out. =
i.J
k..a
k4
i-,
ON
-a
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
[003921 The AUC0-24 hours data (Table 35) shows adjusted values for day 2, 7
and 14. The
baseline PK values (Day 1) are not adjusted and are not relevant given that no
drug was given.
All curves showed a slower onset of absorption and longer exposure curves with
food. Unlike
the impact on C., the impact of food on AUC is complex with some dose levels
showing
higher AUC with fasted conditions (650 mg and 4,875 mg dose), similar AUC
under both fasted
and fed conditions (2925 mg dose) and higher AUC with food (1950 and 6000 mg
dose). The
extension of the curves appear to compensate for the effect of a lower C. peak
for some dose
levels by a wider curve in the 8-16 hour period leading to greater potential
overlap between
doses. For the 6000 mg dose, the AUC is significantly greater (--+46%) when
administered in
the fed state at 4.165 mcg-hr/ml compared with 2.856 fasted. Given that the
6000 mg dose was
administered using the 500 mg tablet, the differences observed may be due to a
subtle effect of
the larger tablet size on the absorption curve, even though in vitro
dissolution data is comparable
for the 325 mg and 500 mg tablets. The larger tablet may have a longer time of
release in vivo.
This difference coupled with food effect delaying the exit of SA from the
stomach may enhance
the overall absorption of SA (acidity should improve sialic acid absorption by
neutralizing its
charge). Therefore all things considered and based on the data, and without
being bound by
theory, providing the larger tablet with food may provide net better drug
absorption. Regardless,
from the results it is clear that either with or without food, adequate
absorption of the drug is
occurring at all dose levels.
[00393} In addition, AUC0-24 hours during Day 14 (or 7th day of repeat dosing)
shows that
overall higher AUC levels can be achieved at steady state when dosing is
divided three times per
day, which suggests that there is some saturation effect on absorption. For
example, at the 6000
mg dose level, the adjusted repeat dose AUCo-24 hours was 6.740 mcg*hr/ml, as
compared with
the AUC levels of 2.856 mcg*hr/m1 (Fasted state) and 4.165 mcg*hr/m1 (Fed
state) at single
doses of 6000 mg. The 6000 mg dose group's AUC is ¨ 235% and ¨160% higher when
the dose
was given as a divided repeat dose rather than as one single dose. The AUC0-24
hours for the 4875
mg repeat dose group was 7.442 mcg*hern1 (Std dev 2.702), which is comparable
to that for
6000 mg repeat dose group at 6.740 mcg*hr/m1 (Std. dev 3.487). Although the
difference in PK
for the 4875 mg and 6000 mg following single dose administration appears
large, the data look
similar following administration of divided doses three times a day, which is
more relevant to the
clinical setting. Given that absorption of SA may be an active pinocytosis-
based process (Oetke
137
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
C., Hinderlich S., Brossmer R., Relater W., Pawlita M., and Keppler CYE 2001.
Evidence for
efficient uptake and incorporation of sialic acid by eukaryotic cells. Eur. J.
Biochem.
268(16):4553-4561), saturation and competition for absorption of drug may be
occurring
simultaneously.
138
Table 35 AUC0-24 hr (mcg x hr/mL) Values Following Single Dose
Administration (Fed and Fasted) and Repeat Doses
of SA-ER
1
c)
b.)
CD
Parameter %:, ,. Statistic 650 m.g.
2925 mg ...
1UC(0-24) N 66
W
.......
.
-
(mcg x 1;rir ) Mean 3.330 :..d.,
3.620 .'.2.:.: ma
CD
SD 0.279 0.459
0.715 0.649 0.329 YP
YP
Median 3.394 3.297
3.868 1.073 3.683 CD
Min, Max 2.939, 3.587
2.861, 4.173 2.404, 4.308 2.627, 4.152 2.968, 3.865
CA
CV(%) 8.383 13.790
19.738 10.080 9.243
Adjusted AUC(0-24) Single/Fasting Day 2 N 6
6 6 4 6
(mcg x hr/m1,1 Mean 1.011 1.393
2.649 4.899 2.856
SD 0.457 0.811
1.451 1.712 1.398
Median 1.058 1.204
2.601 4.185 2.574
Min, Max 0.405, 1.476
0.661, 2.555 0.703, 4.253 3.781, 7.446 1.177, 5.263
CV(%) 45.223 58.244
54.764 34.948 48.951
Single/Fed Day 7 N 6 6
5 4 5
Mean 0.497 1.707
2.651 4.246 4.165
SD 0.339 0.558
0.909 1.501 0.803
Median 0.436 1.733
2.693 4.495 4.406 0
Min, Max 0.195, 1.099
0.971, 2.551 1.314, 3.845 2.249, 5.745 2.940,
4.981 o
m
CV(%) 68.322 32.693
34.303 35.339 19.278 m
I.,
ma
m
W AUC(0-24) Baseline (No Dose) Day i N
8 8 6 6 w
m
YP (mcg x hrimL) Mean 3.437
3.303 3.444 3.554 m
o
SD 0.380
0.631 0.606 0.329 r
m
Median 3.448
3.218 3.412 3.683 1
o
Min, Max 2.939,
4.173 2.404, 4.308 2.627, 4.152 2.968, 3.865 Q
1
cv(%) 11.048
19.094 17.585 9.243 o
m
Adjusted AUC(0-24) Repeat Day 14 N 8
8 4 6
(mcg x hrimi,) Mean 2.407
3.069 7.442 6.740
SD 1.315
0.782 2.702 3.487
Median 2.349
3.250 7.361 6.026
Min, Max 0.594,
4.673 1.831, 4.022 4.788, 10.258 2.511, 11.816
CV(%) 54.625
25.470 36.308 51.728
MO
NOTE: Baseline values for single and repeat dose levels slightly vary because
some patients who are included in one dose group for the single dose phase are
included in a e)
ditTelunt dose group for the repeat dose phase. Therefore, the baseline means
attempt to reflect the mean of the same patients that are being compared, and
the mean changes :71
slightly when you take some patients in or out.
el
C4
b.)
0
=i
GM
......
0
b.)
b.)
wa
CA
-.4
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
[00394] The Tn. data (fable 36) show that food can have a substantial impact
on peak levels,
with the Tn.. shifting much later when drug was administered with food. For
the 4875 mg and
6000 mg dose levels, the shift was from 2.8 hours (Fasted) to 6.0 hours (Fed)
and from 4.3 hours
(Fasted) to 9.6 hours (Fed), respectively. Similar changes are observed at the
other dose levels,
with the exception of the lowest 650 mg dose. The change in peak concentration
time and
overall shape of the curves suggest that food may be an important factor in
absorption of SA-ER.
[003951 The Imax during repeat dosing shows a peak in the evening/nighttime
(i.e., when the
evening dose and the bedtime dose overlap). This timing is positive in terms
of assuring an
adequate level of sialic acid at night during peak protein synthesis.
140
Table 36 Tmax (Hours) Values Following Single Dose Administration
(Fed and Fasted) and Repeat Doses of SA-ER
i
Parameter Vi:, l. : ,.. == . = = = ..
::: 1950 mg = 7.1 48'.' mg (::::. ::., CD
Tmax (hr) Baseline (No Dose) Day 1 N 6
6 6 4 6 b.)
Mean 17.964 16.644
13.344 17.004 9.372 CZ
wa
SD 5.456 8.530
9.356 1.953 8.317 CA)
...,
Median 20.017 19.975
16.033 16.042 9.958 wa
Min, Max 9.000, 23.700
0.000, 23.917 0.000, 24.000 16.000, 19.933 0.000, 20.000
CZ
YP
CV(%) 30.370 51.247
70.113 11.486 89.743 YP
CZ
Ch
Singie/Fasting Day 2 N 6 6
6 4 6
Mean 6.500 3.681
2.856 2.800 4.333
SD 7.036 2.326
1.354 1.499 2.944
Median 4.000 3.025
2.983 2.983 3.000
Min, Max 1.000, 20.000
2.000, 8.000 1.000, 4.167 1.067, 4.167 2.000, 8.000
CV(%) 108.240 63.205
47.426 53.549 67.937
Single/Fed Day 7 N 6 6
5 4 5
Mean 6.347 9.361
4.020 6.004 9.600
SD 2.669 7.438
0.022 2.305 3.578
Median 8.000 8.008
4.017 6.033 12.000
Min, Max 2.000, 8.083
4.000, 24.000 4.000, 4.050 3.950, 8.000 4.000, 12.000
CV(%) 42.049 79.458
0.541 38.391 37.268
0
Baseline (No Dose) Day 1 N 8
8 6 6 o
t.,
Mean 15.473
15.493 18.014 9.372 m
m
SD 7.799
8.653 3.321 9.317
wa
m
4i. Median 17.992
19.975 16.075 9.958 w
m
wa Min, Max
0.000, 23.700 0.000, 23.917 16.000, 24.000 0.000, 20.000
t.,
CV(%) 50.402
55.887 18.436 88.743 o
r
A
I
Repeat Day 14 N 8
8 4 6 0
Mean 19.479
16.513 13.038 17.353 11
o
SD 2.543
9.186 6.805 8.648
Median 20.000
20.000 14.033 20.000
Min, Max 16.000,
24.000 0.000, 24.000 4.083, 20.000 0.000, 24.000
CV(() 13.056
55.632 52.192 49.838
NOTE: Baseline values for single and repeat dose levels slightly vary because
some patients who are included in one dose group for the single dose phase are
included in a
di Ifetent dose group for the repeat dose phase. Theretbre, the baseline means
attempt to reflect the mean of the same patients that are being compared, and
the mean changes
slightly when you take some patients in or out.
mo
e)
tl
el
C4
b.)
0
=i
Go)
....õ
0
b.)
b.)
wa
0
-.4
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
[00396] The terminal half-life (T1/2) of SA-ER at the three higher doses is
similar (-3 hours)
and modestly lengthened by food (Table 37). At the two lowest doses, the T112
appears longer at
5.6 hours (650 mg) and 8.6 hours (1950 mg) but these values do not change
consistently with
food. it is possible that even though the endogenous SA was subtracted in
these adjusted PK
calculations, the 11/7 values were still affected by the presence of an
endogenous pool of S.A of
very similar size, The T112 values for the baseline and repeat dosing are not
meaningful given the
absence of drug or the presence of repeated dosing, respectively.
142
Table 37 T112 (Hours) Values Following Single Dose
Administration (Fed and Fasted) and Repeat Doses of SA-ER
1
Parameter :-:,ase = (-; 650 mg ''.
..g ;:.2. 7,.1 4875 mg 6000 mg C)
T 4.1 (hr) Baseline (No Dose) Day 1 N 2
2 4 3 4 k4
0
Mean 217.709 16.169
112.878 35.404 170.353 wa
SD 8.579 7.823
89.742 11.666 233.409 ua
......
Median 217.709 16.169
110.091 29.624 60.222 wa
0
Min, Max 211.643, 10.637,
21.701 34.520, 27.758, 48.831 40.862,
223.775 196.808 520.105 µ0
0
CV(%) 3.940 48.382
79.504 32.950 137.015 ON
Adjusted T ',2 (hr) Single/Fasting Day 2 N 4
6 6 4 6
Mean 5.581 8.605
3.191 2.687 3.053
SD 3.191 11.576
1.167 0.732 1.300
Median 5.893 3.698
3.469 2.859 3.118
Min, Max 2.144, 8.396
2.856, 32.123 1.123, 4.492 1.657, 3.371 1.440, 4.900
CV(%) 57.175 134.528
36.580 27.254 42.537
Single/Fed Day 7 N 3 6
5 4 5
Mean 8.023 6.206
3.882 3.476 3.418
SD 8.814 3.741
1.299 0.504 0.882
Median 4.033 5.584
4.277 3.509 3.165
Min. Max 1.909, 18.127
1.641, 11.499 1.715, 4.970 2.836, 4.050 2.337, 4.627
0
CV(%) 109.858 60.276
33.470 14.495 25.795 o
r.,
m
m
wg T 11 (hr) Baseline (No Dose) Day 1 N
4 3 4 4 m
co
4. Mean 116.939
138.483 35.568 170.353 w
m
4)
SD 116.552
90.258 9.531 233.409 m
o
Median 116.672
184.122 32.842 60.222 r
m
I
Min, Max 10.637,
34.520, 27.758, 48.831 40.862, o
223.775 196.808 520.105 Q
1
CV(%) 99.669
65.176 26.795 137.015 o
m
Adjusted T '-: (br) Repeat Day 14 N 2
3 2 1
Mean 19.709
33.723 8.733 547.962
SD 8.741
35.060 1.797
Median 19.709
25.351 8.733 547.962
Min, Max 13.528,
25.890 3.606, 72.211 7.463, T.1.003 547.962,
547.962
-V(%) ::4.352
103.966 20.572
MO
e)
tl
NOTE: Baseline values for single and repeat dose levels slightly vary because
some patients who are included in one dose group for the single dose phase are
el
included in a di fferent dose group for the repeat dose phase. Therefore, the
baseline means attempt to reflect the mean of the same patients that are being
uo
k4
compared, and the mean changes slightly when you take some patients in or out.
CD
=i
ua
......
0
k=-)
k=-)
I-.
cr.
--.1
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
Pharmacokinetics Conclusions
[00397} The data show that SA-ER is absorbed and provides steady and
significant drug
levels over a period of 8-16 hours (depending on the dose level) and that
there is definite inter-
patient variability in the degree of SA absorption. The impact of food on AUC
is complex, but
in general, it appears that the C. is lowered and the T. is delayed with food.
The AUC for
the 6000 mg repeat dose was significantly improved with food. At higher dose
levels, mean SA
concentrations reached levels that exceeded normal SA levels by 2x-3x.
[003981 The data at the highest dose of 6000 mg/day did not show greater
absorption than the
4875 mg/day dose level, which may be due to upward skewing of the data in the
smaller 4875
mg single dose cohort in which two out of the four subjects had very high
levels of free SA.
During the single dose administration of 6000 mg/day, the PK parameters
suggest a lower
maximum drug level and a longer PK absorption curve as compared to the 4875
mg/day dose. In
the repeat dosing period (and clinically more relevant setting), the 6000
mg/day dose performed
similar to the 4875 mg/day dose. This is likely due to the longer PK exposure
time of the 500
mg tablet, which led to an overlap between dose events (since dose was
provided three times per
day) and resulted in higher overall AUC levels at that dose.
[00399] Given that the SA levels achieved at all doses are well above the
normal range, and
that the PK curves are relatively steady over the 24 hour cycle, SA-ER tablets
are performing as
expected for an extended release formulation and should achieve levels of free
SA that are
expected to correct the deficiency of sialic acid levels and improve
sialylation in the muscle of
HIBM patients.
Example 9
Combination dose canine studies: Immediate Release Sialic Acid in combination
with
Extended Release Sialic Acid (Tablets)
Preliminary data from canine study showed that Sialic Acid immediate release
formulation (SA-AP!) was better absorbed and gave a higher peak earlier but
was cleared faster.
Thus, the purpose of the combination dose canine studies is to determine
whether the addition of
immediate release sialic acid to Extended Release Sialic Acid (SA-ER) can
provide an increase
in free and total SA levels.
144
CA 02862836 2014-07-02
WO 2013/109906 PCT/US2013/022167
Study 1:
To determine whether the addition of immediate release sialic acid to SA-ER
can
provide an increase in total SA absorbed and mean free SA levels, two
consecutive phases of
dosing were conducted: SA-API (TID) was orally delivered for 4 days followed
by 4 days of
combination dose SA-API+SA.-ER tablets (TID) in dogs. Phase 1: SA-ER tablets:
200
mg/kg/day for 4 days. Phase 2: SA-ER. tablets (200 mg/kg/day) plus SA-API (100
mg/kg/day)
for 4 days. Free sialic acid levels in serum was measured. The results were
shown in Figure 25.
Study 2:
To determine whether the addition of immediate release sialic acid to SA-ER
can
reach steady state after a longer duration of dosing and provide an increase
in total SA.
absorbed/mean free S.A levels, three consecutive phases of dosing were
conducted: S.A-API
(TID) was orally delivered for 4 days followed by 4 days of combination dose
SA-AP14-SA-ER
tablets (TID) and measure free sialic acid levels in serum (dogs). Phase 1: SA-
ER tablets: 200
mg/kg/day for 4 days. Phase 2: SA-ER tablets (200 mg/kg/day) 4- SA-API (100
mg/kg/day) for 4
days. Phase 3: SA-ER. tablets (200 mg/kg/day) 4- SA-API (200 mg/kg/day) for 4
days. Free
sialic acid levels in serum. was measured on Days 4 and 8 and 12 (last day of
each dosing phase).
The results were shown in Figure 26.
Area Under the Curve (AUC) for the end of each phase (Day 4: 0-24hrs):
- TID SA-ER Tablets 200 mg/kg/day: 37.8
- TID SA-ER Tablets 200 mg/kg/day + SA-API 100 mg/kg/day: 66.4
- TID SA-ER Tablets 200 mg/kg/day + SA-API 200 mg/kg/day: 90.4
Conclusion: Oral combination of SA-ER tablets and SA-API boosts overall
absorption
than SA-ER tablets alone. Thus, with 50% more drug in immediate release form,
the free sialic
acid levels in the serum can be doubled.
[004001 All publications and patent applications cited in this
specification are incorporated
herein by reference as if each individual publication or patent application
were specifically and
individually indicated to be incorporated by reference.
145
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[00401] Although the foregoing invention has been described in some detail
by way of
illustration and example for purposes of clarity of understanding, it is
apparent to those skilled in
the art that certain minor changes and modifications will be practiced.
Therefore, the description
and examples should not be construed as limiting the scope of the invention.
146
