U.S. patent application number 14/973465 was filed with the patent office on 2016-04-14 for compositions and methods for treating spinocerebellar ataxia.
The applicant listed for this patent is BIOBLAST PHARMA LTD.. Invention is credited to Dalia MEGIDDO.
Application Number | 20160101122 14/973465 |
Document ID | / |
Family ID | 51865231 |
Filed Date | 2016-04-14 |
United States Patent
Application |
20160101122 |
Kind Code |
A1 |
MEGIDDO; Dalia |
April 14, 2016 |
COMPOSITIONS AND METHODS FOR TREATING SPINOCEREBELLAR ATAXIA
Abstract
The present invention provides alleviating a sign or symptom of
spinocerebellar ataxia by administering an injection of an aqueous
formulation comprising trehalose.
Inventors: |
MEGIDDO; Dalia; (Nataf,
IL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
BIOBLAST PHARMA LTD. |
Tel Aviv |
|
IL |
|
|
Family ID: |
51865231 |
Appl. No.: |
14/973465 |
Filed: |
December 17, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14818863 |
Aug 5, 2015 |
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14973465 |
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14665648 |
Mar 23, 2015 |
9125924 |
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14818863 |
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14320184 |
Jun 30, 2014 |
9084720 |
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14665648 |
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PCT/IL2014/050411 |
May 7, 2014 |
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14320184 |
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61990027 |
May 7, 2014 |
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61820278 |
May 7, 2013 |
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Current U.S.
Class: |
514/53 |
Current CPC
Class: |
A61K 31/7016 20130101;
A61K 9/0019 20130101 |
International
Class: |
A61K 31/7016 20060101
A61K031/7016 |
Claims
1. A method of alleviating a sign or symptom of spinocerebellar
ataxia (SCA) by administering to a subject in need thereof an
injection of an aqueous formulation comprising a single active
ingredient consisting of trehalose, wherein the formulation (i) has
a pH about 4.5 to 7.0, (ii) contains less than 0.75 endotoxin units
per mL, and (iii) is administered at a per administration dose of
between 5 to 50 grams trehalose.
2. The method of claim 1, wherein the formulation is administered
once weekly.
3. The method of claim 1, where the formulation is administered at
a per administration dose of about 0.5 gram trehalose per kilogram
body weight.
4. The method of claim 1, wherein the per day dose is 8, 15 or 30
grams.
5. The method of claim 1, wherein the rate of administration is
such that the maximum endotoxin level is less than 5 EU per
kilogram of body weight per hour.
6. The method of claim 1, wherein the trehalose is at 10%
(w/v).
7. The method of claim 1, wherein the formulation has an osmolality
of about 280-330 mOsm/kg.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 14/818,863, filed Aug. 5, 2015. U.S. patent
application Ser. No. 14/818,863 is a continuation of Ser. No.
14/665,648, filed Mar. 23, 2015, now U.S. Pat. No. 9,125,924,
issued Jul. 16, 2015. U.S. patent application Ser. No. 14/665,648
is a continuation of U.S. patent application Ser. No. 14/320,184,
filed Jun. 30, 2014, now U.S. Pat. No. 9,084,720, issued Jun. 30,
2015. U.S. patent application Ser. No. 14/320,184 claims priority
to and benefit of U.S. Provisional Application No. 61/990,027,
filed May 7, 2014. U.S. patent application Ser. No. 14/320,184 is a
continuation in part of PCT/IL2014/050411, filed May 7, 2014, which
claims priority to and benefit of U.S. Provisional Application No.
61/820,278, filed May 7, 2013. The contents of the aforementioned
patent applications are each herein incorporated by reference in
their entireties.
FIELD OF THE INVENTION
[0002] This invention relates generally intravenous compositions of
trehalose for the treatment of signs and symptoms of aggregation
disease or disorders including oculopharyngeal muscular dystrophy
(OPMD).
BACKGROUND OF THE INVENTION
[0003] Oculopharyngeal Muscular Dystrophy (OPMD) is a rare
inherited myopathy characterized by ptosis, severe dysphagia and
proximal limb weakness. Its estimated prevalence is 1:100,000 and
the largest clusters reported were in families of French-Canadians
origin in Canada and in the US (prevalence 1:1000), Bukhara Jews in
Israel (prevalence 1:600) and Hispanics in New Mexico, Arizona
Colorado and California [1-4]. OPMD is inherited, in most cases, as
an autosomal dominant trait with complete penetrance. The disease
is equally prevalent among both genders. The gene associated with
the disease has been identified. This mutation results in
production of an abnormal poly (A) binding protein nuclear 1
protein (PABPN 1), a nuclear protein involved in pre-mRNA
polyadenylation, transcription regulation, and mRNA
nucleocytoplasmic transport.
[0004] The disease is most often diagnosed in the fifth-sixth
decades of life and progresses throughout the patient's life. By
age 70 the majority of patients suffer from all or some of the
following symptoms: severe dysphagia ptosis, tongue atrophy and
weakness, lower and upper limb proximal weakness, dysphonia,
limitation in upward gaze and facial muscle weakness. As ptosis
becomes more pronounced patients adapt the "astronomer posture,"
tilting of the head and upward gaze--further aggravating the
dysphagia. The dysphagia starts with difficulty in swallowing solid
food and progresses to liquids as well. As the dysphagia becomes
more severe, patients become malnourished, cachectic, dehydrated
and suffer from repeated aspiration pneumonia. OPMD does not seem
to shorten life expectancy but is associated with severe
debilitation and reduced quality of life.
[0005] There is no medical treatment or potential cure for OPMD.
Current therapeutic strategies are confined to surgical
interventions aimed at alleviating ptosis. Repeated cricopharyngeal
dilatations are frequently used to relieve dysphagia. Myotomy of
the upper esophageal sphincter muscles has also been employed.
These procedures may provide only temporary relief and do not
affect the progression of the disease that eventually leads to
severe difficulty in swallowing, recurrent aspiration with
increasing risk of aspiration pneumonia and severe weight loss
which are the most common causes of mortality in OPMD patients.
[0006] Accordingly, there is an urgent need for compositions and
therapeutic methods for alleviating the signs and symptoms of
oculopharyngeal muscular dystrophy.
SUMMARY OF THE INVENTION
[0007] The present invention is based upon the discovery that rapid
intravenous injection of trehalose results in cellular uptake and
retention of trehalose. Trehalose has been shown to prevent
pathological aggregation of proteins within cells. Thus, the
present invention provides an aqueous formulation for intravenous
injection, comprising a single active ingredient consisting of
trehalose and methods of use thereof.
[0008] In one embodiment, the trehalose formulation of the present
invention has a pH about 4.5 to 7.0. In another embodiment, the
formulation contains less than 0.75 endotoxin units per mL. In
another embodiment, the formulation contains 10% (w/v) trehalose.
In another embodiment, the formulation has an osmolality of about
280-330 mOsm/kg.
[0009] The trehalose formulation of the present invention is
retained in the cell for an extended period of time after
intravenous administration of the formulation to the cell. In one
embodiment, the formulation is retained in the cell for about 48-72
hours after intravenous injection. In another embodiment, the
intravenous injection is completed within 120 minutes.
[0010] The trehalose formulation of the present invention prevents
aggregation of proteins within the cells. In one embodiment, the
formulation of the present invention prevents aggregation of a
protein that is involved in the pathogenesis of or associated with
a sign or symptom of oculopharyngeal muscular dystrophy
spinocerebellar ataxia (SCA). In a further embodiment, the
formulation of the present invention prevents aggregation of the
abnormal protein PABPN1.
[0011] The present invention also provides a method of alleviating
a sign or symptom of a disease, by intravenously administering to a
subject in need thereof a formulation of the present invention. In
one embodiment, the intravenous injection of the formulation is
completed within 120 minutes. In another embodiment, the
formulation is administered once weekly. In another embodiment, the
formulation is administered at 0.5 gram trehalose per kilogram body
weight per day. In another embodiment, the formulation is
administered between 5 to 35 grams trehalose per day. In another
embodiment, the disease is oculopharyngeal muscular dystrophy. In a
further embodiment, the sign or symptom includes, but is not
limited to, muscle weakness, formation of protein aggregates (e.g.,
PABPN1 aggregates), and formation of pathological skeletal muscle
fibers.
[0012] Other features and advantages of the invention will be
apparent from and are encompassed by the following detailed
description and claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0013] FIG. 1 Mean Plasma Concentrations (ng/mL) of Trehalose
Following Single Intravenous or Oral Administration of Trehalose
Dihydrate at 1000 mg/kg to Male SD Rats.
[0014] FIG. 2 Mean Plasma and Muscle Concentrations (ng/mL or ng/g)
of Trehalose Following Single Intravenous Administration of
Trehalose Dihydrate at 1000 mg/kg to Male SD Rats.
[0015] FIG. 3 Mean Plasma and Muscle Concentrations (ng/mL or ng/g)
of Trehalose Following Single Oral Administration of Trehalose
Dihydrate at 1000 mg/kg to Male SD Rats.
[0016] FIG. 4 Plasma Glucose Ratios of Drug Treated Group to That
of Vehicle Group Following Intravenous or Oral Administration of
Vehicle or Trehalose Dihydrate at 1000 mg/kg to Male SD Rats.
[0017] FIG. 5 Mean Plasma Glucose Levels (mmol/L) Following
Intravenous or Oral Administration of Vehicle or Trehalose
Dihydrate at 1000 mg/kg to Male SD Rats.
[0018] FIG. 6 Individual Plasma Glucose Levels (mmol/L) Following
Intravenous Administration of Vehicle to Male SD Rats.
[0019] FIG. 7 Individual Plasma Glucose Levels (mmol/L) Following
Oral Administration of Vehicle to Male SD Rats.
[0020] FIG. 8 Mean Plasma Glucose Levels (mmol/L) Following
Intravenous Administration of Trehalose Dihydrate at 1000 mg/kg or
Vehicle to Male SD Rats.
[0021] FIG. 9 Mean Plasma Glucose Levels (mmol/L) Following Oral
Administration of Trehalose Dihydrate at 1000 mg/kg or Vehicle to
Male SD Rats.
DETAILED DESCRIPTION
[0022] The present invention provides aqueous solutions of
trehalose for intravenous injection. Surprisingly, rapid
intravenous administration of trehalose results in cellular uptake
and retention 48-72 hours. Accordingly, the invention provides
methods for treating the signs and symptoms of oculopharyngeal
muscular dystrophy (OPMD) by intravenous administration or
trehalose. In other aspects, the compositions and methods disclosed
herein may be used to treat other aggregation disease or disorders
such as poly-alanine aggregation disorder, poly-glutamine
aggregation disorder and a tauopathy. In particular aspects, the
disease or disorder is spinocerebellar ataxias (SCA), Friedreich's
ataxia, spinal and bulbar muscular atrophy (SBMA), Huntington's
disease, Parkinson's disease, Alzheimer's disease and amyotrophic
lateral sclerosis (ALS), dentatorubral-pallidoluysian atrophy
(DRPLA), Pick's disease, Corticobasal degeneration (CBD),
Progressive supranuclear palsy (PSP) and Frontotemporal dementia
and parkinsonism linked to chromosome 17 (FTDP-17).
[0023] Before describing the present invention in detail, it is to
be understood that unless otherwise indicated, this invention is
not limited to particular dosages, formulations or methods of use,
as such may vary. It is also to be understood that the terminology
used herein is for the purpose of describing particular embodiments
only, and is not intended to be limiting.
[0024] It must be noted that, as used in this specification and the
appended claims, the singular forms "a," "an" and "the" include
plural referents unless the context clearly dictates otherwise.
Thus, for example, "a dosage form" refers not only to a single
dosage form but also to a combination of two or more different
dosage forms, "an active agent" refers to a combination of active
agents as well as to a single active agent, and the like.
[0025] As used in the specification and the appended claims, the
terms "for example." "for instance," "such as," "including" and the
like are meant to introduce examples that further clarify more
general subject matter. Unless otherwise specified, these examples
are provided only as an aid for understanding the invention, and
are not meant to be limiting in any fashion.
[0026] Unless defined otherwise, all technical and scientific terms
used herein have the meaning commonly understood by one of ordinary
skill in the art to which the invention pertains. Although any
methods and materials similar or equivalent to those described
herein may be useful in the practice or testing of the present
invention, preferred methods and materials are described below.
Specific terminology of particular importance to the description of
the present invention is defined below.
[0027] When referring to a compound of the invention, applicants
intend the term "compound" to encompass not only the specified
molecular entity but also its pharmaceutically acceptable,
pharmacologically active analogs, including, but not limited to,
salts, polymorphs, esters, amides, prodrugs, adducts, conjugates,
active metabolites, and the like. When the term "compound" is used,
then, it is to be understood that applicants intend to include that
compound per se as well as pharmaceutically acceptable,
pharmacologically active salts, polymorphs, esters, amides,
prodrugs, adducts, conjugates, metabolites, and other such
derivatives, analogs and related compounds.
[0028] The terms "treating" and "treatment" as used herein refer to
reduction in severity and/or frequency of signs or symptoms,
elimination of signs or symptoms and/or underlying cause,
prevention of the occurrence of symptoms and/or their underlying
cause (e.g., prophylactic therapy), and improvement or remediation
of damage.
[0029] By the terms "effective amount" and "therapeutically
effective amount" of a compound of the invention is meant a
nontoxic but sufficient amount of the drug or agent to provide the
desired effect.
[0030] Efficaciousness of treatment is determined in association
with any known method for diagnosing or treating OPMD. Alleviation
of one or more signs or symptoms of OPMD indicates that the
compound confers a clinical benefit.
[0031] The term "injection" as used herein refers to a bolus
injection, slow bolus injection over several minutes, or prolonged
infusion, or several consecutive injections/infusions that are
given at spaced apart intervals. Generally, since rate of
administration of the disclosed injectable formulation is also
determined in consideration of the level of endotoxins in the
formulation, a per administration "injection" of an effective dose
of the trehalose can be divided into spaced apart injections of
lower amounts of formulation, until the whole of the effective dose
is administered. Such spaced apart injections per a single
administration are also referred to herein as "per administration"
or "per administration injection" or the like, or in other words, a
single administration can include several injections or prolonged
infusion. The administration of the aqueous injectable solution of
trehalose, particularly for the treatment of OPMD, as disclosed
herein is completed in no more than 120 minutes, and the rate of
administration is such that the maximum endotoxin level is no more
than 5 EU per kilogram of body weight of the patient per hour.
[0032] 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 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.
[0033] By "patient" is meant any animal for which treatment is
desirable. Patients may be mammals, and typically, as used herein,
a patient is a human individual.
[0034] The term "about" as used herein indicates values that may
deviate up to 1%, more specifically 5%, more specifically 10%, more
specifically 15%, and in some cases up to 20% higher or lower than
the value referred to, the deviation range including integer
values, and, if applicable, non-integer values as well,
constituting a continuous range.
Compositions
[0035] "Trehalose" is a stable, nonreducing disaccharide with two
glucose molecules linked in a 1,1 configuration.
[0036] Trehalose is well known for its protein-stabilizing
properties [10,11]. It is used extensively in many applications as
a stabilizer of frozen food, in freeze-drying of biological systems
and cells, as a stabilizer of therapeutic parenteral proteins and
as an excipient in tablets and IV solutions. Trehalose is
recognized as a GRAS (Generally Regarded as Safe) food ingredient
by the FDA and is listed on the USP-NF (United States Pharmacopoeia
National Formulary), EP (European Pharmacopoeia) and JP (Japanese
Pharmacopoeia). This disaccharide chemical chaperone has been shown
to prevent pathological aggregation of proteins within cells in
several diseases associated with abnormal cellular-protein
aggregation such as Huntington's disease, spinocerebellar ataxia,
Parkinson and Alzheimer disease.
[0037] OPMD is characterized by intracellular aggregation of the
abnormal protein PABPN1 by products of which are considered by most
authorities toxic [12]. Trehalose was found effective in reducing
the aggregation and toxicity of mutant PABPN1 proteins in OPMD cell
models. Furthermore, treatment of an OPMD mouse model with
trehalose resulted in the attenuation of muscle weakness, decreased
aggregate formation and a reduced number of pathological skeletal
muscle fibers [13]. As such, it was hypothesized that trehalose
might be useful for treatment of OPMD.
[0038] Like all disaccharides, trehalose is metabolized at the
epithelial brush border to two D-glucose molecules. Less than 0.5%
of ingested trehalose is absorbed into the blood stream where it is
further metabolized by liver and kidney by trehalase. Oral
trehalose in amounts exceeding 40-50 gram per day causes diarrhea
and bloating. Thus in order to achieve therapeutic amounts of
trehalose in the muscle cells it was necessary to circumvent the
massive metabolism in the GI tract. Therefore the inventors
developed an I.V. solution of trehalose.
[0039] To date, the safety and toxicity of trehalose has been
extensively investigated, and the substance was found to be safe
when administered both orally and intravenously, in doses that are
substantially higher than the intended therapeutic dose.
[0040] Accordingly, in one embodiment, the compositions described
herein comprise trehalose as the sole active agent. Furthermore, in
one embodiment, the methods described herein comprise the
intravenous administration of trehalose to a patient in need
thereof.
[0041] The compositions of the current disclosure comprise, as an
active agent, trehalose in a pharmaceutically acceptable form. The
active agent, trehalose, may be administered in the form of the
compound per se, as well as in the form of a salt, polymorph,
ester, amide, prodrug, derivative, or the like, provided the salt,
polymorph, ester, amide, prodrug or derivative is suitable
pharmacologically. Salts, esters, amides, prodrugs and other
derivatives of the active agents may be prepared using standard
procedures known to those skilled in the art of synthetic organic
chemistry and described, for example, by J. March. Advanced Organic
Chemistry: Reactions, Mechanisms and Structure, 4th Ed. (New York:
Wiley-Interscience, 1992). For any active agents that may exist in
enantiomeric forms, the active agent may be incorporated into the
present compositions either as the racemate or in enantiomerically
pure form.
[0042] For compositions administered as aqueous or other
solvent-based dosage forms (e.g., for parenteral administration), a
variety of liquid carriers may be used. Aqueous solutions may
include salts, buffers, and the like.
[0043] Salts are compounds that ionize in aqueous solutions and may
be employed, for example, to adjust the tonicity of the solution.
If the active agent is present in the form of a salt, additional
salts may be added to the composition in order, for example, to
effect ion exchange with the active agent. Salts suitable for use
with the compositions described herein are known in the art and
include, for example, lithium, sodium, potassium, calcium, and
magnesium salts having appropriate counterions that may be selected
from chloride, bromide, iodide, carbonate, phosphate, nitrate,
silicate, sulfate, phosphite, nitrite, sulfite, and the like.
[0044] Buffers are compounds or solutions that are employed to aid
in maintaining the concentration of an analyte within a desired
range. For example, pharmaceutically acceptable pH buffers are used
to maintain the acidity or basicity of a solution within a
pharmaceutically acceptable range. Buffers for use in the
compositions disclosed herein may be any known or hereafter
discovered buffer.
[0045] Excipients are inactive ingredients that may be employed in
the compositions described herein for a variety of reasons. A wide
range of excipients are described in the literature (e.g., Rowe et
al., Handbook of Pharmaceutical Excipients. McGraw Hill, 2006).
[0046] The amount of trehalose in the compositions disclosed herein
will depend on a number of factors and will vary from subject to
subject. Such factors include the severity of the symptoms, the
patient's age, weight and general condition, and the judgment of
the prescribing physician.
[0047] Optimally the pH of the formulation is about 4.5 to 7.0. The
osmolality of the formulation is about 280-330 mOsm/kg.
[0048] The formulation contains less than 1.0, 0.9, 0.8, 0.75, 0.7,
0.6, 0.5, 0.4, 0.3, 0.2, 0.1 or less endotoxin units per mL.
[0049] Preferably the aqueous formulation is about 50%, 40%, 30%,
20%, 10%, 5% or less trehalose (w/v).
[0050] The formulation of claim 1, wherein the formulation has an
osmolality of about 280-330 mOsm/kg.
[0051] In one embodiment, the purified trehalose is substantially
free of contaminants resulted from the protein used in the
enzymatic preparation process of the trehalose, such as organic
solvents used in the process, e.g., ammonium, acetonitrile,
acetamide, alcohol (e.g., methanol, ethanol, or isopropanol), TFA,
ether or other contaminants. In this context "substantially" free
of contaminants means that the contaminant content of the peptide
at the end of the purification process is preferably less than
0.5%, less than 0.3%, less than 0.25%, less than 0.1%, less than
0.05%, less than 0.04%, less than 0.03%, less than 0.02%, less than
0.01%, less than 0.005%, less than 0.003%, or less than 0.001% of
the total weight of the trehalose. The content of contaminants can
be determined by conventional methods such as gas
chromatography.
[0052] Preferably, the residual solvents in the purified trehalose
of the invention are less than the limits set in the ICH
guidelines, e.g., IMPURITIES: GUIDELINE FOR RESIDUAL SOLVENTS
Q3C(R5) (available at
http://ww.ich.Org/fileadmin/Public_Web_Site/CH_Products/Guidelines/Qualit-
y/Q3C/Step4/Q 3C_R5_Step4.pdf). For example, the purified trehalose
contains <5000 ppm ethanol (e.g., <140 ppm), and/or <3000
ppm methanol.
Methods of Use
[0053] The compositions and methods described herein are useful in
the treatment of the signs and symptoms of OPMD. Signs and symptoms
of OPMD include severe dysphagia, ptosis, tongue atrophy and
weakness, lower and upper limb proximal weakness, dysphonia,
limitation in upward gaze and facial muscle weakness.
[0054] In one embodiment, a method is provided for treating a
patient suffering from OPMD. The methods of treatment involve
administering a therapeutically effective amount of a composition
comprising trehalose to the patient. Administration of trehalose
may be carried out using any of the compositions, modes of
administration, and dosage forms described herein.
Dosages
[0055] Pharmaceutical formulations suitable for use in conjunction
with the present disclosure include compositions wherein trehalose
is contained in a "therapeutically effective" amount, i.e., in an
amount effective to achieve its intended purpose, such as treatment
of OPMD. Determination of a therapeutically effective amount is
well within the capability of those skilled in the art.
[0056] Toxicity and therapeutic efficacy of the compositions
described herein can be determined by standard pharmaceutical
procedures in cell cultures or experimental animals, e.g.,
procedures used for determining the maximum tolerated dose (MTD),
the ED.sub.50, which is the effective dose to achieve 50% of
maximal response, and the therapeutic index (TI), which is the
ratio of the MTD to the ED.sub.50. Obviously, compositions with
high TIs are the most preferred compositions herein, and preferred
dosage regimens are those that maintain plasma levels of the
trehalose at or above a minimum concentration to maintain the
desired therapeutic effect. Dosage will, of course, also depend on
a number of factors, the site of intended delivery, the route of
administration, frequency of administration, and other pertinent
factors known to the prescribing physician. The dosage range may be
from each of 10, 20, 50, 75, 100, 150, 200, 300 mg/Kg body weight
per day up to each of 400, 450, 500, 550, 600, 650, 700, 750, 800,
850, 900, 950 and 1000 mg/Kg body weight per day. Generally,
however, dosage will be in the range of approximately 0.1
grams/kg/day to 1 g/kg/day. Preferably the dose is less than 0.54
grams/kg/day.
[0057] In some embodiments the trehalose is administered such that
the total daily dose (on a day of administration) is between about
5 grams to 50 grams. In preferred embodiments the total per
administration dose of trehalose is 8, 15 or 30 grams. In
particular embodiments the trehalose is administered as a single
dose of 5, 8, 15, 30, 40 or 50 grams.
[0058] In certain aspects, the dosing regimen is equal doses. In
other aspects, gradually increasing doses, or gradually decreasing
doses may be used. For example, in certain aspects, a subsequent
dose may be greater or lesser than a prior dose by about 10%, 20%,
30%, 40%, 50%, or about 100%.
Administration
[0059] Administration is accomplished such that that the maximum
endotoxin level is less than 5 EU per kilogram of body weight per
hour. In particular aspects, the endotoxin level is less than about
1, 2, 3, or less than about 4 endotoxin units per kilogram of body
weight per hour.
[0060] Administration is daily, weekly, biweekly or monthly.
Preferably, the administration is weekly.
[0061] Administration of the compositions described herein may be
carried out as part of a treatment regimen that may include
multiple instances of administration of trehalose-containing
compositions as well as administration of other pharmaceutically
active compositions. Such a regimen may be designed as a method of
treatment for OPMD, and/or as a method of long-term maintenance of
the health of a patient after having been treated for OPMD (e.g.,
preventing recurrences). The treatment regimen may be designed as a
method of treating a subject that is asymptomatic for OPMD, that is
a subject that has been genetically diagnosed with OPMD but does
not have any symptoms. Such treatment regimen will delay the onset
of OPMD symptoms in a subject. It will be appreciated that
determination of appropriate treatment regimens is within the skill
of practitioners in the art.
[0062] Administration of the compositions described herein may be
carried out using any appropriate mode of administration and dosage
form. Preferably administration is parenteral. The term
"parenteral" as used herein is intended to include, for example,
subcutaneous, intravenous, and intramuscular injection. Most
preferably, the administration is intravenous Over 99.5% of the
trehalose is not absorbed into the blood stream. In addition, oral
amounts of trehalose higher than 50 g a day in humans frequently
cause diarrhea, bloating and discomfort. Thus, in particular
aspects, the trehalose may be administered as an intravenous as an
aqueous formulation to address poor absorption into the bloodstream
and minimized undesirable metabolic events. In specific
embodiments, the pH of the formulation is about 4.5 to 7.0, the
osmolality of the formulation is about 280-330 mOsm/kg, the
formulation contains less than 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4,
0.3, 0.2, 0.1 or less endotoxin units per mL and the aqueous
formulation is about 50%, 40%, 30%, 20%, 10%, 5% or less trehalose
(w/v).
[0063] The trehalose may be delivered over a suitable period. In
some embodiments administration is complete within from about 75 to
about 120 minutes, specifically within less than 90 minutes.
[0064] In certain embodiments, effective serum levels of trehalose
are achieved within from about 10 to about 20 or 30 or 40 or 50 or
60 minutes following trehalose administration. In certain
embodiments, effective serum levels of the active ingredient are
achieved within from about 5 to about 20 or 30 or 40 or 50 or 60
minutes following trehalose administration. In certain embodiments,
effective serum levels of the active ingredient are achieved within
from about 20 to about 20 or 30 or 40 or 50 or 60 minutes following
trehalose administration. In certain embodiments, effective serum
levels of the active ingredient are achieved within about 5, 10,
15, 20, 30, 40, 50 or 60 minutes following trehalose
administration.
Delivery Systems
[0065] Administration of trehalose for medical uses requires safe
and efficient delivery systems. The present disclosure provides
delivery systems (e.g. formulations for parenteral administration)
for safe delivery of a variety of substances due to their special
physico-chemical features. The delivery systems significantly
enhance efficiency and quality of trehalose absorption based on its
unique physicochemical features, which enables lower concentrations
or amounts of active substance to be delivered to a subject in a
biologically active form. The present delivery systems provide for
the direct access of the active substance to the tissues and thus
provide immediate or near-immediate effects of trehalose to the
subject in need thereof.
[0066] Accordingly, in certain embodiments, the present invention
provides a pharmaceutical delivery system for the improved
administration of trehalose or physiologically active derivative
thereof, comprising as the active ingredient said trehalose or
physiologically active derivative thereof in a suitable carrier for
fast restoration of relief of symptoms of the disease of the
treated subject.
[0067] In certain embodiments, the drug delivery systems may
provide the active substance in a controlled release mode. In
certain embodiments, the drug delivery systems of the invention may
further comprises at least one additional pharmaceutically active
agent.
[0068] The presently disclosed delivery systems can generally
comprise a buffering agent, an agent which adjusts the osmolality
thereof, and optionally, one or more pharmaceutically acceptable
carriers, excipients and/or additives as known in the art.
Supplementary pharmaceutically acceptable active ingredients can
also be incorporated into the compositions. The carrier can be
solvent or dispersion medium suitable for
parenterally-administrable compositions containing, for example,
water, ethanol, polyol (for example, glycerol, propylene glycol,
and liquid polyethylene glycol, and the like), suitable mixtures
thereof, and vegetable oils. The proper fluidity can be maintained,
for example, by the use of a coating, such as lecithin, by the
maintenance of the required particle size in the case of dispersion
and by the use of surfactants.
[0069] As indicated above, the present trehalose delivery system
can be administered in controlled-, sustained- or delayed-release
formulations. Any controlled or sustained release method known to
those of ordinary skill in the art may be used with the
formulations and methods of the presently disclosed subject matter
such as those described in Langer 1990 [16]. Such method comprises
administering a sustained-release composition or a coated
implantable medical device so that a therapeutically effective dose
of the composition of the invention is continuously delivered to a
subject of such a method. Sustained release may also be achieved
using a patch designed and formulated for the purpose. Controlled
or sustained-release compositions include formulation in lipophilic
depots (e.g., fatty acids, waxes, oils). Also comprehended by the
invention are particulate compositions coated with polymers (e.g.,
poloxamers or poloxamines). Sustained release formulae or devices,
or any topical formulations, may additionally contain compositions
to stabilize the composition or permeate physiological barrier such
as skin or mucous membrane. Exemplary additional components may
include any physiologically acceptable detergent, or solvent such
as, for example, dimethylsulfoxide (DMSO).
[0070] In certain embodiments, the trehalose in the present
compositions can be formulated for sustained or controlled release
over a period of at least 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or
12 hours. In certain embodiments, the trehalose in the present
compositions can be formulated for sustained or controlled release
over a period of about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12
hours. In certain embodiments, the trehalose in the present
compositions can be formulated for sustained or controlled release
over a period of between about 0.5 or 1 or 2 or 3 or 4 hours and
about 5, 6, 7, 8, 9, 10, 11 or 12 hours. In certain embodiments,
the trehalose in the present compositions can be formulated for
sustained or controlled release over a period of between about 5 or
6 or 7 or 8 hours and about 9, 10, 11 or 12 hours.
[0071] In certain embodiments, the trehalose in the present
compositions can be in immediate, fast of burst release form.
[0072] In certain embodiments, the trehalose in the present
compositions can be formulated to release up to 5, 10, 15, 20, 25,
30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99, 99.5 or
100% of the total trehalose in about 0.5, 1, 2, 3, 4, 5, 6, 7 or 8
hours. In certain embodiments, the trehalose in the present
compositions can be formulated to release not less than 5, 10, 15,
20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99,
99.5 or 100% of the total trehalose in about 0.5, 1, 2, 3, 4, 5, 6,
7 or 8 hours.
[0073] In certain embodiments, the trehalose in the present
compositions can be in a combination of sustained or slow release
and immediate or fast or burst release forms. In certain
embodiments, the relative proportion of sustained or slow release
trehalose to immediate or fast release trehalose is. e.g., 1 to 99,
5 to 95, 10 to 90, 15 to 85, 20 to 80, 25 to 75, 30 to 70, 35 to
65, 40 to 60, 45 to 55, 50 to 50, 55 to 45, 60 to 40, 65 to 35, 70
to 30, 75 to 25, 80 to 20, 85 to 15, 90 to 10, 95 to 5, or 99 to
1.
[0074] In certain embodiments, a polymeric material is used to
sustain or control release of trehalose. In certain embodiments,
the type of polymeric material and the amount of which is used,
have a strong influence on the rate of release of trehalose from
the present compositions and delivery systems. Examples of polymers
include both hydrophobic and hydrophilic polymers. Examples of
hydrophobic polymers include, but are not limited to, ethyl
cellulose and other cellulose derivatives, fats such as glycerol
palmito-stearate, beeswax, glycowax, castorwax, carnaubawax,
glycerol monostearate or stearyl alcohol, hydrophobic
polyacrylamide derivatives and hydrophobic methacrylic acid
derivatives, as well as mixtures of these polymers. Hydrophilic
polymers include, but are not limited to, hydrophilic cellulose
derivatives such as methyl cellulose, hydroxypropylmethyl
cellulose, hydroxyethylcellulose, hydroxypropyl cellulose,
carboxymethyl cellulose, sodium carboxymethylcellulose and
hydroxyethyl methylcellulose polyvinyl alcohol, polyethylene,
polypropylene, polystyrene, polyacrylamide, ethylene vinyl acetate
copolymer, polyacrylate, polyurethane, poly vinylpyrrolidone,
polymethylmethacrylate, polyvinyl acetate, polyhydroxyethyl
methacrylate, as well as mixtures of these polymers. Furthermore,
any mixture of one or more hydrophobic polymer and one or more
hydrophilic polymer could optionally be used.
[0075] The trehalose contained in the present compositions and
delivery systems may be entrapped in liposomes, micro- and
nano-particles.
[0076] In certain embodiments, a polymeric material to be used in
the present compositions and delivery systems is microcrystalline
cellulose such as "Avicel PH 101" manufactured by FMC BioPolymer's.
Alternatively, a polymeric material to be used in the present
compositions and delivery systems is hydroxypropyl methylcellulose
such as "Metholose" produced by Shin-Etsu Chemical Co. In certain
embodiments, a polymeric material to be used in the present
compositions and delivery systems is ethyl cellulose such as
"Ethocel.TM." manufactured by The Dow Chemical Company. In certain
embodiments, a polymeric material to be used in the present
compositions and delivery systems is an acrylic polymer such as
"Eudragit RS.TM." produced by Rohm GmbH. In certain embodiments, a
polymeric material to be used in the present compositions and
delivery systems is a colloidal silicone dioxide such as
"Aerosil.TM." manufactured by Degussa. In certain embodiments, a
polymeric material to be used in the present compositions and
delivery systems is a Poly (Vinyl Acetate) such as "Kollicoat SR"
manufactured by BASF. In certain embodiments, a polymeric material
to be used in the present compositions and delivery systems is an
ethyl acetate and vinyl acetate solution such as "Duro-Tak"
manufactured by Delasco Dermatologic Lab & Supply, Inc.
[0077] In certain embodiments, delivery systems of the invention
comprise delivery devices. In certain embodiments, the compositions
of the invention are delivered by an osmotic process at a
controlled rate such as by an osmotic pump. The system may be
constructed by coating an osmotically active agent with a rate
controlling semipermeable membrane. This membrane may contain an
orifice of critical size through which agent is delivered. The
dosage form after coming into contact with aqueous fluids, imbibes
water at a rate determined by the fluid permeability of the
membrane and osmotic pressure of the core formulation. This osmotic
inhibitions of water result in formation of a saturated solution of
active material with in the core, which is dispensed at controlled
rate from the delivery orifice in the membrane.
[0078] In certain embodiments, the compositions of the invention
are delivered using biodegradable microparticles. In certain
embodiment, the system to prepare microparticles consists of an
organic phase comprised of a volatile solvent with dissolved
polymer and the material to be encapsulated, emulsified in an
aqueous phase. In certain embodiments, the biodegradable polymers
that can be used for the microparticle matrix, comprises polylactic
acid (PLA) or the copolymer of lactic and glycolic acid (PLAGA).
The PLAGA polymer degrades hydrolytically over time to its
monomeric components, which are easily removed from the body
through natural life processes.
[0079] The preparation may also contain an absorption enhancer and
other optional components. Examples of absorption enhancers
include, but are not limited to, are cyclodextrins, phospholipids,
chitosan, DMSO, Tween, Brij, glycocholate, saponin, fusidate and
energy based enhancing absorption equipment.
[0080] Optional components present in the dosage forms include, but
are not limited to, diluents, binders, lubricants, surfactants,
coloring agents, flavors, buffering agents, preservatives,
stabilizing agents and the like.
[0081] Diluents, also termed "fillers," include, for example,
dicalcium phosphate dihydrate, calcium sulfate, lactose, cellulose,
kaolin, mannitol, sodium chloride, dry starch, hydrolyzed starches,
silicon dioxide, colloidal silica, titanium oxide, alumina, talc,
microcrystalline cellulose, and powdered sugar. For administration
in liquid form, the diluents include, for example, ethanol,
sorbitol, glycerol, water and the like.
[0082] Binders are used to impart cohesive qualities to the
formulation. Suitable binder materials include, but are not limited
to, starch (including corn starch and pregelatinized starch),
gelatin, sugars (including sucrose, glucose, dextrose, lactose and
sorbitol), polyethylene glycol, waxes, natural and synthetic gums,
e.g., acacia, tragacanth, sodium alginate, celluloses, and Veegum,
and synthetic polymers such as polymethacrylates and
polyvinylpyrrolidone.
[0083] Lubricants are used to facilitate manufacture: examples of
suitable lubricants include, for example, magnesium stearate,
calcium stearate, stearic acid, glyceryl behenate, and polyethylene
glycol.
[0084] Surfactants may be anionic, cationic, amphoteric or nonionic
surface active agents, with anionic surfactants preferred. Suitable
anionic surfactants include, but are not limited to, those
containing carboxylate, sulfonate and sulfate ions, associated with
cations such as sodium, potassium and ammonium ions. Particularly
preferred surfactants include, but are not limited to: long alkyl
chain sulfonates and alkyl aryl sulfonates such as sodium
dodecylbenzene sulfonate; dialkyl sodium sulfosuccinates, such as
sodium bis-(2-ethylhexyl)-sulfosuccinate; and alkyl sulfates such
as sodium lauryl sulfate.
[0085] Stabilizing agents such as antioxidants, include, but are
not limited to, propyl gallate, sodium ascorbate, citric acid,
calcium metabisulphite, hydroquinone, and 7-hydroxycoumarin. If
desired, the present compositions may also contain minor amounts of
nontoxic auxiliary substances such as wetting or emulsifying
agents, preservatives, and the like.
[0086] The invention will be further described in the following
examples, which do not limit the scope of the invention described
in the claims.
EXAMPLES
Example 1
Analysis of Trehalose Dihydrate
[0087] Two (2) samples were brought in for analysis on Feb. 17,
2014.
[0088] The samples were kept at room temperature until
analysis.
Sample identification:
TABLE-US-00001 Analyst number Manufacturer Lot number Expiry date
14-01025 Hayashibara 3G281 27 Jul. 2016 14-01026 Pfanstiehl 34943A
August 2015
[0089] The samples were analyzed according to NF 31 page 2266-2267
during Analyst study 2014-003 "Trehalose NF Determination of Assay
and Related Substances by HPLC Method Verification". Specificity
was demonstrated for glucose and maltotriose as required by the
monograph. The specificity for trisaccharides produced by enzymatic
modification of starch was not in the scope of the study. Sample
14-01026 was analyzed in six replicates and sample 14-01025 was
analyzed in duplicate.
[0090] Results:
TABLE-US-00002 Sample 14-01025 Sample 14-01026 Hayashibara
Pfanstiehl Test Specifications lot 3G281 lot 34943A Assay
97.0-102.0% w/w on 98.2% (conforms) 99.4% (conforms) anhydrous
basis Related Maltotriose Conforms Conforms substances .ltoreq.0.5%
w/w Any peak eluting Conforms Conforms before trehalose
.ltoreq.0.5% w/w Glucose .ltoreq.0.5% w/w Conforms Conforms Any
peak eluting after Conforms Conforms trehalose .ltoreq.0.5% w/w
Example 2
Caballeta for IV Injection Certificate of Analysis
TABLE-US-00003 [0091] TABLE 2 Analysis of Trehalose Formulation
Test Specification Result Appearance of container Clear glass 30R
vial with grey rubber Clear glass 30R vial with grey ATP007
stopper, aluminum seal and white flip rubber stopper, aluminum seal
and off lid. white flip off lid. Appearance of contents Clear
colourless liquid essentially free Clear colourless liquid
essentially ATP007 from visible particulate matter free from
visible particulate matter P537 Identity Retention time of the P537
peak .+-.5% Retention time of the P537 ATP1323 of standard peak is
within .+-.5% of the P537 standard peak P537 Content Assay 90.0% to
110.0% label claim 99.8% ATP1323 P537 Related Substances Report
individual impurities Maltotriose: None detected (% label claim)
.gtoreq.0.05% label claim Unknown RRT0.90: 0.1% ATP1323 Report
total impurities Glucose: None detected Maltotriose and other
polysaccharides Total Impurities: 0.1% eluting before P537:
.ltoreq.0.5% Glucose and peaks eluting after P537 .ltoreq.0.5%
Total impurities s 2.0% pH 4.5 to 7.0 6.2 ATP164 Osmolality 280-330
mOsm/kg 289 mOsm/kg ATP841 Particulate matter Particulates
.gtoreq.10 .mu.m: NMT 6000 .gtoreq.10 .mu.m: 7 USP <788>, Ph.
Eur Particulates .gtoreq.25 .mu.m: NMT 600 .gtoreq.25 .mu.m: 0
2.9.19 Extractable Volume Not less than 30 ml 32 ml USP <1>,
Ph. Eur 2.9.17 Endotoxins <0.24 EU/ml Point 1 tray 1: <0.1
EU/ml USP <85>, Ph. Eur 2.6.14 Point 4 tray 31: <0.1 EU/ml
Point 8 tray 72: <0.1 EU/ml Sterility Complies No growth USP
<71>, Ph. Eur 2.6.1
Example 3
Preclinical PK Study
[0092] The plasma and muscle concentrations of trehalose in male
Sprague-Dawley (SD) rats was determined after intravenous bolus
(IV) and oral gavage (PO) administration.
[0093] All applicable portions of the study confirmed to the
following regulations and guidelines regarding animal care and
welfare: AAALAC International and NIH guidelines as reported in the
"Guide for the Care and Use of Laboratory Animals," National
Research Council ILAR, Revised 1996.
[0094] The study included 42 SD rats (male, 250 to 350 grams in
weight, the Shanghai SLAC Laboratory Animal Co. Ltd.). Animals were
administered with a volume of 5 ml/kg trehalose formulation
(trehalose dihydrate in sterilized water at 200 mg/mL) to achieve a
nominal dose of 1 gr/kg, intravenously or orally.
[0095] Blood samples were collected after each dose administration
and processed for plasma. Muscle samples (hind leg muscle) were
collected and homogenized. The concentrations of trehalose in
plasma and muscle homogenate samples were analyzed by qualified
bioanalytical LC/MS/MS methods.
Pharmacokinetics Data Analysis
[0096] Plasma concentration data of trehalose was subjected to a
non compartmental pharmacokinetic analysis using WinNonlin software
program (version 6.3, Pharsight, Mountain View, Calif.). Zero-time
intercept concentration (CO), volume of distribution (Vdss),
Clearance (Cl), peak plasma concentrations (Cmax) and the
corresponding peak times (Tmax), terminal half-life (T1/2), mean
residence time (MRT) from time zero to the last time point
(MRT0-last), MRT from time zero to infinity (MRT0-inf), the area
under the plasma concentration-time curve (AUC) from time zero to
the last time point (AUC0-last) and AUC from time zero extrapolated
to infinity (AUC0-inf) were calculated using the linear/log
trapezoidal rule. Nominal sampling times were used to calculate all
pharmacokinetic parameters since there was not any deviation larger
than 5% between the actual and nominal sampling times.
[0097] The values of muscle to plasma concentration and AUC ratio
(M/P ratio) were both calculated.
Trehalose Concentration in Plasma and Muscle
[0098] Pharmacokinetic parameters of trehalose in the plasma and
muscle following single intravenous or oral administration of
trehalose dihydrate solution (200 mg trehalose dihydrate per 1 mL
sterilized water) at 1000 mg/kg to male SD rats are presented in
Table 3 below.
[0099] Individual and mean plasma concentrations of trehalose
following intravenous or oral administration of trehalose dihydrate
solution (200 mg trehalose dihydrate per 1 mL sterilized water) at
1000 mg/kg to male SD rats are presented in Table 3 and shown
graphically in FIG. 1.
[0100] Individual and mean muscle concentrations of trehalose
following single intravenous or oral administration of trehalose
dihydrate solution (200 mg trehalose dihydrate per 1 mL sterilized
water) at 1000 mg/kg to male SD rats are presented in Table 3 as
well. Plasma and muscle concentrations comparison for trehalose
following single intravenous or oral administration of trehalose
dihydrate at 1000 mg/kg to male SD rats are nd shown graphically in
FIGS. 2 to 3.
[0101] Following a single intravenous dose of trehalose solution
(200 mg trehalose dihydrate per 1 mL sterilized water) at 1000
mg/kg to male fasted SD rats in tested groups 1 to 5, trehalose
showed a total clearance (CI) of 17.2 mL/min/kg (approximately
31.3% of rat liver blood flow (=55 mL/min/kg)), with the averaged
elimination half life (T.sub.1/2) of 2.07 hours. The Co was
1,370,000 ng/mL.
[0102] The volume distribution (V.sub.dss) was at 0.685 L/kg. The
mean plasma exposure AUC.sub.0-last (48 hr) was 778,000
nghr/mL.
[0103] With an oral administration of trehalose dihydrate solution
(200 mg trehalose dihydrate per 1 mL sterilized water) to male SD
rats in tested groups 6 to 10, trehalose maximum plasma
concentration (C.sub.max=4,280 ng/mL) was attained at 0.5 hour post
dose (T.sub.max). The AUC.sub.0-last (3 hr) was 4,520 ng/mLhr. The
absolute bioavailability of trehalose was estimated to be as low as
0.601%.
[0104] The pharmacokinetic properties of trehalose demonstrated a
rapid absorption with a time to reach peak plasma concentrations,
but the absolute oral bioavailability was very low, which noted
that the compound may undergo a significant presystemic
metabolism.
[0105] Following a single intravenous dose of trehalose dihydrate
solution (200 mg trehalose dihydrate per 1 mL sterilized water) at
1000 mg/kg to male SD rats in tested groups 1 to 5, the C.sub.max
of trehalose in muscle was 3730 ng/mL, which was observed at 8
hours (T.sub.max) post dose. The muscle exposure AUC.sub.0-last (48
hr) was 107,000 nghr/mL with the elimination half-life 33.8 hours.
The PK parameters of muscle samples in drug treated oral group
(Groups 6 to 10) could not be calculated because they were below
LLOQ
[0106] The mean ratios of muscle trehalose concentration to plasma
concentration ranged from 2.88 to 3.76 in male SD rats following
intravenous administration. Muscle to plasma concentration ratios
for trehalose upon oral administration were below LLOQ and not
calculable.
CONCLUSIONS
[0107] Following intravenous or oral administrations of trehalose
dihydrate solution in sterilized water at 1000 mg/kg to male SD
rats, trehalose in plasma and muscle tissue were determined. Plasma
glucose was also monitored for each sample from study animals of
drug treated groups. The following conclusions can be made:
[0108] First, following IV administration, the total clearance (Cl)
of trehalose was 17.2 mL/min/kg, accounting for approximately 31.3%
of liver blood flow, a moderate value of hepatic extraction ratio.
The V.sub.dss and T.sub.1/2 were 0.685 L/kg and 2.07 hours
respectively. The mean plasma exposure AUC.sub.0-last was 778,000
nghr/mL.
[0109] Following oral administration, trehalose demonstrated a
rapid absorption with T.sub.max observed at 0.50 hours post dose,
but the absolute oral bioavailability was as low as 0.601%,
suggesting presystemic metabolism may play an important role.
T.sub.1/2 of trehalose was markedly shortened in oral
administration rats in comparison to the intravenous group.
[0110] Following IV administration, the mean ratios of muscle
trehalose concentration to plasma concentration ranged from 2.88 to
3.76 in male SD rats.
[0111] Finally, it was observed that trehalose dihydrate was well
tolerated by the rats at the given dosage.
TABLE-US-00004 TABLE 3 Pharmacokinetic parameters Matrix Plasma
Muscle Group ID IV PO IV PO C.sub.0 (ng/mL) 1370000 -- -- ND
C.sub.max (ng/mL or ng/g) -- 4280 3730 ND T.sub.max (h) -- 0.500
8.00 ND T.sub.1/2 (h) 2.07 0.740 33.8 ND CI (mL/min/kg) 17.2 -- --
ND V.sub.dss (L/kg) 0.685 -- -- ND AUC.sub.0-last (ng h/mL or
778000 4520 107000 ND ng h/g) AUC.sub.0-inf (ng h/mL or 781000 4870
183000 ND ng h/g) MRT.sub.0-last (h) 0.618 1.04 21.1 ND
MRT.sub.0-inf (h) 0.666 1.26 52.6 ND AUC.sub.0-inf/AUC.sub.0-last
100 108 171 ND (%) .sup.cBioavailability (%) -- 0.601 -- --
.sup.dAUC ratio -- -- 0.234 ND Abbreviations: ND = Not determined;
.sup.cBioavailability (%) was calculated with mean AUC.sub.0-inf
and nominal dose; .sup.dAUC Ratio = Muscle AUC.sub.0-inf/Plasma
AUC.sub.0-inf; AUC.sub.(0-inf) >120% of AUC.sub.(0-last).
[0112] Thus, as demonstrated in Table 3, the trehalose in the IV
administered formulation showed T.sub.1/2 of 2.07 hour in plasmas,
over two-fold higher than the plasma T.sub.1/2 obtained for
trehalose in the orally administered formulation (0.740 hours). In
addition, the muscle T.sub.1/2 obtained for the trehalose in the IV
administered formulation was 33.8 hours. The AUC values obtained
for plasma and muscle when the formulation was administered IV were
also significantly higher than the respective AUC values of
formulation administered orally.
[0113] In addition, as demonstrated in FIG. 1, the mean plasma
concentration of the trehalose in the IV administered formulation
is higher than the mean plasma concentration of the trehalose in
the orally administered formulation at each of the tested time
points.
[0114] Interestingly, FIG. 2, which demonstrates plasma versus
muscle concentrations of trehalose for a trehalose formulation
administered intravenously, shows that muscle concentrations are
higher than plasma concentrations of trehalose. Plasma and muscle
concentrations of trehalose were undetectable for a trehalose
formulation administered orally.
Summary of Pre-Clinical Study
TABLE-US-00005 [0115] Study Details Administered compound Trehalose
Sponsor BBP Sponsor Study No. NA Wuxi DMPK Study No.
BBP-20140220-RPK Species Male SD Rat, fasted Study Group IV PO
Nominal dose (mg/kg) 1000 1000 Administered dose (mg/kg 804 778
Formulation IV 200 mg/mL in water for injection, clear solution
Formulation PO 200 mg/mL in water for injection, clear solution
In-life start date Mar. 11, 2014
TABLE-US-00006 Bioanalytical Details Analyte compound Trehalose
Batch No. 34943A Molecular weight 342.3 Formular weight 378.33 Salt
factor 1.11 Purity 90.5 Analytical technique LC-MS/MS Matrix Plasma
(EDTA-K.sub.2)
TABLE-US-00007 PK Calculation Settings Program Phoenix WinNonlin
6.3 Model IV-Noncompartmental model 201 (intravascular input)
PO-Noncompartmental model 200 (extravascular input) Calculation
Linear/log trapezoidal method Dose used (WNL Nominal dosage "BQL"
was excluded in the PK parameters and mean plasma concentration
calculation.
TABLE-US-00008 Comments 1. The value of all the samples "predose"
was below the lower limit of quantitation (LLOQ = 100 ng/mL) 2.
Stock solution for standard curve was dissolved in DMSO with active
content considered at 81.9%. However, our protocol calculated
active content at 90.5%. Finally, all the measured value was
adjusted with following. Actual concentration value = Measured
value/0.905
Example 4
Trehalose Disposition
[0116] Oral absorption: When ingested orally, most of the sugar is
not assimilated as a disaccharide into the blood stream. Rather, it
is enzymatically hydrolyzed in the small intestine by a
trehalose-specific disaccharidase (trehalase) into two d-glucose
molecules, which are subsequently absorbed and metabolized.
Trehalase is found in most animals at the brush border of the
intestinal mucosa, as well as in the kidney, liver and plasma.
Although trehalose does not occur in mammalian cells, humans have
the enzyme trehalose in intestinal villae cells and in kidney brush
border cells, probably to handle ingested trehalose [11,14,15].
[0117] Biotransformation and excretion: When trehalose enters blood
circulation it is rapidly converted to glucose by trehalase in
serum, kidney, liver and bile, depending on the species. In animals
lacking trehalase activity in the kidney such as rats, most of the
intravenous trehalose is excreted in the urine, proportional to
plasma concentration. In animals where renal trehalase activity
exists (such as guinea-pigs and rabbits), only a very small
fraction of trehalose is recovered in the urine. When rabbits were
given 500 mg of trehalose intravenously (corresponding to
200.+-.300 mg/kg body weight) the compound was cleared from the
plasma within 60 min, and none was detected in the urine [15].
Example 5
Animal Safety
[0118] The safety and tolerability of trehalose has been
extensively investigated. A detailed review of safety in animals
and humans is presented in the Cabaletta Investigator's brochure,
incorporated herein by reference (the term Cabaletta as used herein
signifies a 10% IV solution of trehalose).
[0119] Animal toxicity: Trehalose LD50 was examined in mice, rats
and dogs. Neither species showed any signs of toxicity and no
deaths occurred after oral and intravenous administration. The
results are summarized in Table 1:
TABLE-US-00009 TABLE 1 LD50 of trehalose in animals SPECIES ROUTE
LD50 (mg/kg bw) Mouse Oral >5000 Mouse Intravenous >1000 Rat
Oral >16000 Rat Oral >5000 Rat Intravenous >1000 Dog Oral
>5000 Dog Intravenous >1000
[0120] Human Safety and Use:
[0121] Trehalose is recognized as a safe food ingredient as well as
a GRAS material used in the pharmaceutical industry as an excipient
for oral, intraocular and I.V. drug formulations.
[0122] In several studies, healthy volunteers were given oral doses
of trehalose ranging from 10 to 60 gr. Apart from mild abdominal
symptoms (e.g. flatulence, distension, borborygmus and occasional
diarrhea) no other safety issues were reported.
[0123] Trehalose has been used as a protein stabilizer in several
commercially available protein drugs for over a decade and its
safety has repeatedly been established in patient populations at
advanced stages of malignant diseases, hemophilia and related
clotting disorders. These drugs are approved for use for several
years, and are sometimes given to patients as frequently as every 8
hours through 2-3 weeks intervals.
Example 6
Clinical Study
[0124] A three-center, multi-national, randomized, double-blind,
dose escalation and parallel-group dose-controlled study will be
conducted to assess the safety, tolerability, and efficacy of IV
Cabaletta.RTM. in Patients with Oculopharyngeal Muscular Dystrophy
(OPMD). The study will be comprised of an Exploratory phase
(screening period and a treatment period), an interim analysis, and
a Pivotal phase (second treatment period and a follow-up
period).
[0125] Up to 30 adult patients with OPMD will be enrolled into the
study at each of the three study sites. A minimum of 42 patients
will be enrolled in total.
[0126] Inclusion Criteria
1. Males and females 2. 18-80 years (inclusive) of age 3.
Genetically diagnosed with OPMD 4. Moderate dysphagia (abnormal
drinking test at screening and on the first dosing day, before drug
administration) 5. Patients must be ambulatory, and capable of
performing the muscle functional and strength assessments 6.
Patients who provide written informed consent to participate in the
study 7. Body Mass Index (BMI)<30 kg/m2 8. Female patients of
child-bearing potential must have a negative serum pregnancy test
at screening 9. Male and females must agree to use acceptable birth
control 10. Patients must be able to understand the requirements of
the study and be willing to comply with the requirements of the
study
[0127] Exclusion Criteria
1. Diabetes mellitus Type 1 or 2 2. Other major diseases, e.g.
renal failure (creatinine clearance <60 ml/min), liver failure
and chronic liver diseases (e.g. hepatitis B or C), HIV carriers,
tuberculosis, SLE, rheumatoid polyarthritis, sarcoidosis,
collagenosis 3. Uncontrolled heart disease, e.g., CHF 4. Other
neuromuscular diseases 5. Other disorders associated with
esophageal dysphagia: e.g. gastroesophageal reflux (GERD),
esophageal stricture due to mechanical or chemical trauma,
infection (e.g. esophageal moniliasis), drug-induced dysphagia
(e.g. bisphosphonates), esophageal rings and webs, spastic motility
disorders of the esophagus. 6. History of malignancy 7. History of
neck irradiation 8. Pregnant or currently lactating women 9.
Obesity (BMI.gtoreq.30) and associated morbidity 10. Prior
pharyngeal myotomy 11. Weight loss of more than 10% in the last 12
months. 12. Known hypersensitivity to any ingredients in the
injection
[0128] Dose
[0129] Cabaletta.RTM., a 10% IV solution of trehalose, will be
administered once a week for 72 weeks. Study drug will be delivered
over approximately 80 minutes. Doses used in the study:
TABLE-US-00010 Dose Study week Initial dose (unblinded) 8 g Week 1
Second dose (unblinded) 15 g Week 2 treatment ( 30 g Week 3 to week
24 Randomized 30 g or no treatment Week 25-72, discontinuation
[0130] Study Objectives
[0131] Exploratory Phase (24 Weeks) Study Objectives
[0132] Primary
[0133] 1. To determine the safety and tolerability of Cabaletta in
OPMD patients after a single (8 gr) IV administration.
[0134] 2. To determine the safety and tolerability of Cabaletta in
OPMD patients after a single (15 gr) IV administration
[0135] 3. To determine the safety and tolerability and 30 g IV
Cabaletta in OPMD patients after repeated weekly dosing.
[0136] 5. To obtain data on the pharmacokinetics of trehalose.
[0137] 6. To determine the pharmacokinetics of trehalose (Israel
site only)
[0138] Secondary
[0139] 1. To determine the effect of Cabaletta on the progression
of OPMD, assessed by measuring dysphagia, swallowing-related
quality of life (SWAL-QOL), and muscle function and strength.
[0140] 2. At the end of the exploratory phase patients will be
randomized into 2 grouped: 1 group will continue with the weekly
injections of 30 grams and the other will not get treatment however
they will monitored through the pivotal phase in the same way the
treated patients will be.
[0141] Pivotal Phase (48 Weeks) Study Objectives
[0142] Primary
[0143] 1. To determine the effect of Cabaletta on the progression
of OPMD, assessed by measuring dysphagia, swallowing-related
quality of life (SWAL-QOL), and muscle function and strength
[0144] 2. To compare the efficacy 30 g Cabaletta with no
treatment.
[0145] Secondary
[0146] 1. To determine the safety and tolerability of 30 g IV
Cabaletta in OPMD patients after repeated weekly dosing.
[0147] 2. The study will be comprised of an Exploratory phase
(screening period and a treatment period), an interim analysis,
blinded randomization and a Pivotal phase (second treatment period
and a follow-up period) as follows:
[0148] Study Procedures
[0149] Exploratory Phase
[0150] Screening Period (Week -4/Day -28 to Week 0/Day 0)
[0151] Screening assessments will be conducted over two visits
within 28 days prior to the start of therapy, as specified in the
Schedule of Assessments.
[0152] Treatment Period 1 (Week 1 to Week 24)
[0153] All eligible patients will receive study treatment once a
week.
Initially, all eligible patients will receive one dose of Cabaletta
8 g over one week, followed by 15 g Cabaletta over the next week
(Visits 3 and 4). If no safety concerns arise, at Visit 5/Week 3
all patients will receive to receive Cabaletta 30 g for 24 weeks.
The first 4 infusions will be done at the clinic under the
direction of the study investigator. The patients must return to
the clinic once a month for drug infusion and study assessments, as
indicated in the Schedule of Procedures: all other weekly infusions
may be done in the patient's home or in the clinic.
[0154] Interim Analysis
[0155] The interim analysis will be conducted when the first
patient enrolled into the study has completed 6 months of therapy,
and the last subject enrolled has completed at least 3 months of
therapy. Both safety and efficacy will be examined in the interim
analysis.
[0156] Pivotal Phase
[0157] Treatment Period 2 (Week 25 to Week 72)
[0158] Patients Will be Randomized into Two Arms: Continued
Treatment Arm and Discontinuation Arm (No Treatment Control)
[0159] Patients assigned to the continued treatment arm will
continue weekly IV infusions of Cabaletta at home or at the
hospital, except for Visits 32, 40, 48, 60 and 72. Study procedures
will be done as specified in the Schedule of Assessments.
[0160] Patients assigned to the no-treatment control will not be
getting additional infusion however they will be followed up and
monitored in same schedule planned for the treatment arm,
[0161] Follow-Up Period (4 Weeks Post-Dose)
[0162] Patients will be seen at a post-treatment follow-up visit
(Visit 75), 4 weeks after the final dose.
[0163] Safety and Tolerability Outcome and Assessments
[0164] The primary safety endpoint is the frequency, severity, and
duration of adverse events (AEs), including clinically significant
laboratory abnormalities after administration of Cabaletta.
[0165] Safety will be evaluated on the basis of the following
assessments:
[0166] AEs and concomitant medications: Continuous (starting from
informed consent signature until end of study)
[0167] Physical examination: on screening (Visit 1), on the first 4
dosing visits, and then once monthly in the experimental phase, at
the interim analysis and once every 6-12 weeks at the pivotal phase
as outlined in the schedule of assessment in appendix A.) and End
of Study (EOS, Visit75).
[0168] 12-lead ECG: at screening (Visit 1), during the first dosing
with 15 g and 30 g (Visits 4 and 5), at the interim analysis (Visit
26), and at the end of study evaluation (Visit 75).
[0169] Vital signs: on screening (Visit 1) and on each visit until
end of study. On the first 4 dosing visits, vital signs will be
assessed prior to the Cabaletta administration, every 30 min during
administration and 30 min following the drug administration.
[0170] Safety laboratory evaluations will be conducted according to
the Schedule of Procedures. Evaluations will include: complete
blood count (CBC) with differential, electrolytes (Na, K, Cl). BUN,
creatinine, glucose, liver function tests (ALT, AST, total
bilirubin, direct bilirubin, alkaline phosphatase, and serum
albumin), and dipstick urinalysis.
[0171] Pre- and post-dose blood glucose will be measured at the
first 3 dosing visits; blood glucose will also be assessed at the
End of Study evaluation (Visit 75).
Urine pregnancy will be done at Screening (Visit 1), baseline
(Visit 3), and EOS (Visit 75).
[0172] Safety data will be reviewed periodically by an independent
data safety monitoring board.
[0173] Evaluation
[0174] The following assessments will be performed: [0175]
Penetration Aspiration Score (using Videofluoroscopy) [0176]
SWAL-QOL [0177] Muscle timed functional and strength
assessments
[0178] Changes compared to baseline will be measured for each
patient, and the total change in scores for the treatment groups in
each pre-determined efficacy endpoint will be statistically
analyzed.
[0179] The following additional assessments, thought to be
supportive in nature, will be performed at the times specified in
the Schedule of Assessments: [0180] Weight [0181] Drinking test
[0182] Percutaneous Core Needle Biopsy (PCNB) will be performed to
obtain muscle fiber for histology.
[0183] Pharmacokinetics
[0184] The pharmacokinetic of trehalose will be assessed in
patients (Israeli site only) at the randomized dose of 15 g or 30 g
Cabaletta. Trehalose blood concentration will be measured pre-dose
(up to 60 minutes before study drug administration); and every 30
minutes after dosing is initiated, for 5 hours or until glucose
levels return to normal, whichever occurs first.
[0185] Statistical Methods
[0186] All measured variables and derived parameters will be listed
individually and, if appropriate, tabulated by descriptive
statistics. For descriptive statistics summary tables will be
provided giving sample size, absolute and relative frequency and
95% Confidence Interval (CI) for categorical variables and sample
size, arithmetic mean, standard deviation, coefficient of variation
(if appropriate), median, minimum and maximum, percentiles and 95%
CI for means of continuous variables.
[0187] Rate of subjects with any adverse event and potentially
clinically significant laboratory results with 95% CI will be
calculated by treatment period.
Exploratory statistical analyses of data may be undertaken as
appropriate.
[0188] All tests applied will be two-tailed, and p value of 5% or
less will be considered statistically significant.
TABLE-US-00011 Schedule of Assessments Screening and Assessment
Baseline Exploratory Phase Pivotal Phase End of Study Visit No. 1 2
3 4 5 6 7 8 9 10 11-13 14 15-17 18 19-21 22 23-25 26 27-33 34 35-41
42 43-61 62 63-73 74 75 Dosing Week -4 1 2 3 4 5 6 7 8 9-11 12
13-15 16 17-19 20 21-23 24 25-31 32 33-39 40 41-59 60 61-71 72 --
Informed Consent X -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- --
-- Interim Analysis -- -- -- -- -- -- -- -- -- Demographics X -- --
-- -- -- -- -- -- -- -- -- -- -- -- -- -- -- and randomuzaion -- --
-- -- -- -- -- -- -- Medical history X -- -- -- -- -- -- -- -- --
-- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- Weight X -- X --
-- X -- -- -- X -- X -- X -- X -- X -- X -- X -- X -- X X Vital
signs X X X X X X X X X X X X X X X X X X X X X X X X X X X
Physical X -- X X X X -- -- -- X -- X -- X -- X -- X -- X -- X -- X
-- X X examination 12-Lead ECG X -- -- X X -- -- -- -- -- -- -- --
-- -- -- -- X -- -- -- -- -- -- -- -- X Safety labs X -- X X X X --
-- -- -- -- X -- -- -- -- -- -- -- X -- X -- X -- X X Urine
pregnancy X -- X -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- --
-- -- -- -- -- -- X Blood glucose -- X X X -- -- -- -- -- -- -- --
-- -- -- -- -- -- -- -- -- -- -- -- -- X HIV, HBsAg, X -- -- -- --
-- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- --
HCVAb Drinking test X -- X -- -- X -- -- -- X -- X -- X -- X -- X
-- X -- X -- X -- X X Inclusion/Exclusion X -- X -- -- -- -- -- --
-- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- --
Videofluoroscopy -- X -- -- -- -- -- -- -- -- -- -- -- -- -- -- --
X -- -- -- -- -- -- -- -- X SWAL-QOL -- X -- -- -- -- -- -- -- --
-- -- -- -- -- -- -- X -- -- -- -- -- -- -- -- X Muscle -- X -- --
-- -- -- -- -- -- -- -- -- -- -- -- -- X -- -- -- -- -- -- -- -- X
function/strength Muscle Biopsy -- X -- -- -- -- -- -- -- -- -- --
-- -- -- -- -- X -- -- -- -- -- -- -- -- X Randomization -- -- --
-- X -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- -- --
-- Dosing at study site -- -- X X X X X -- X -- X -- X -- X -- X --
X -- X -- X -- Dosing at patient's -- -- -- -- -- -- X X X -- X --
X -- X -- X -- X -- X -- X -- X -- -- home or study site Adverse
Events -- X X X X X X X X X X X X X X X X X X X X X X X X X X
Concomitant X X X X X X X X X X X X X X X X X X X X X X X X X X X
Medications Pharmacokinetics -- -- -- -- -- -- X -- -- -- -- -- --
-- -- -- -- -- -- -- -- -- -- -- -- -- -- (Israel site only)
Example 7
Determination of Endotoxin Level
[0189] It is accepted that the maximal allowed level of endotoxin
in formulations administered intravenously is 5 endotoxin units
(EU) per kg body mass per hour (5 EU/kg/hr). In order to determine
the theoretical maximum endotoxin level IV per kg body mass/hour
(K) in trehalose formulation (solution of trehalose dihydrate in
sterilized water), the following calculations were made:
TABLE-US-00012 TABLE 4 Calculation of maximal endotoxin levels in
trehalose formulations Endotoxin 15 gr trehalose 30 gr trehalose
contribution formulation formulation 2.4 EU/gr (trehalose) 36 72
0.5 EU/ml (solvent) 75 (in 150 ml) 150 (in 300 ml) Total EU in
formulation 111 222 Assuming 75 min infusion 88.8 EU/hr 177.6 EU/hr
K for 60 kg body weight 1.5 3.0 K for 50 kg body weight 1.8 3.6 K
for 40 kg body weight 2.2 4.4
[0190] As indicated in Table 4 above, endotoxin level per ml in
trehalose formulations prepared with standard solvents (e.g. water,
saline, etc.) is 0.74 EU/ml. Assuming a moderate infusion rate of
75 minutes, for a body weight of 60, 50 and 40 kg the endotoxin
level in trehalose 10% (w/v) formulations is 1.5, 1.8 and 2.2
EU/kg/hr, respectively, for a formulation comprising 15 gr
trehalose and 3.0, 3.6 and 4.4 EU/kg/hr, respectively, for a
formulation comprising 30 gr trehalose.
[0191] Accordingly, under the maximum rate planned, the endotoxin
level for a body weight of 60, 50 and 40 kg will be 2, 2.4 and 3
EU/kg/hr, respectively, for a formulation comprising 15 gr
trehalose (in 150 ml solvent) and 4, 4.8 and 6 EU/kg/hr,
respectively, for a formulation comprising 30 gr trehalose (in 300
ml solvent).
OTHER EMBODIMENTS
[0192] While the invention has been described in conjunction with
the detailed description thereof, the foregoing description is
intended to illustrate and not limit the scope of the invention,
which is defined by the scope of the appended claims. Other
aspects, advantages, and modifications are within the scope of the
following claims.
* * * * *
References