U.S. patent application number 13/660642 was filed with the patent office on 2013-05-09 for enzyme delivery systems and methods of preparation and use.
The applicant listed for this patent is Joan M. Fallon, Matthew Heil. Invention is credited to Joan M. Fallon, Matthew Heil.
Application Number | 20130113129 13/660642 |
Document ID | / |
Family ID | 42934578 |
Filed Date | 2013-05-09 |
United States Patent
Application |
20130113129 |
Kind Code |
A1 |
Fallon; Joan M. ; et
al. |
May 9, 2013 |
Enzyme Delivery Systems and Methods of Preparation and Use
Abstract
This invention relates to coated digestive enzyme preparations
and enzyme delivery systems and pharmaceutical compositions
comprising the preparations. This invention further relates to
methods of preparation and use of the systems, pharmaceutical
compositions and preparations to treat persons having ADD, ADHD,
autism, cystic fibrosis and other behavioral and neurological
disorders.
Inventors: |
Fallon; Joan M.;
(Bronxville, NY) ; Heil; Matthew; (Sherman,
CT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Fallon; Joan M.
Heil; Matthew |
Bronxville
Sherman |
NY
CT |
US
US |
|
|
Family ID: |
42934578 |
Appl. No.: |
13/660642 |
Filed: |
October 25, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13407408 |
Feb 28, 2012 |
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13660642 |
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12386051 |
Apr 13, 2009 |
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13407408 |
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Current U.S.
Class: |
264/15 |
Current CPC
Class: |
A61P 1/00 20180101; A61K
9/1075 20130101; C12Y 301/01003 20130101; A61K 9/5015 20130101;
A61P 25/28 20180101; A61K 9/1682 20130101; A61J 15/00 20130101;
A61K 9/167 20130101; A61K 38/48 20130101; A61K 9/009 20130101; C12Y
304/21001 20130101; A61P 3/00 20180101; C12Y 302/01 20130101; A61P
43/00 20180101; A61K 9/16 20130101; A61K 9/1617 20130101; A23V
2002/00 20130101; A61K 9/5063 20130101; A61K 38/47 20130101; A61K
38/54 20130101; A61P 25/00 20180101; A61P 25/16 20180101; A61P
25/18 20180101; A61P 25/14 20180101; Y02A 50/30 20180101; A61K
38/4826 20130101; A61K 38/465 20130101; A61K 9/0053 20130101; A23L
33/17 20160801; A61K 38/48 20130101; A61K 2300/00 20130101; A61K
38/47 20130101; A61K 2300/00 20130101; A61K 38/465 20130101; A61K
2300/00 20130101 |
Class at
Publication: |
264/15 |
International
Class: |
A61K 9/16 20060101
A61K009/16 |
Claims
1-59. (canceled)
60. A method of producing a pellet or a micropellet consisting of
pancreatin, which pellet contains no auxiliary substances and
binding agents, comprising the steps of: a.) comminuting the
pancreases of pigs or cattle and conducting autolysis; b.)
obtaining a sieved filtrate by filtration of the intermediate
product obtained in step a); c.) precipitating the enzymes from the
sieved filtrate; d.) filtering the mixtures obtained in step c) to
obtain a filter cake; e.) grinding and vacuum-drying the filter
cake until a residual moisture of 0.1 to 0.3% by weight is
obtained, whereby the pancreatin product with 0.1 to 0.3% by weight
of residual moisture constitutes the dried end product; whereas the
extrudable filter cake mass contains more residual moisture and/or
organic solvent residues, which are of the order of 50%; f.)
thermally treating the filter cake at 80.degree. C. or at a
temperature below 80.degree. C.; g.) extruding the thermally
treated filter cake having sufficient plasticity and having no
additives and/or binding agents in order to form strands; and h.)
spheronising the extrusion of step g) without additives and/or
binding agents or auxiliary substances so as to obtain spherical,
elliptical or drop-shaped pellets or micropellets.
61. The method of claim 60, wherein the auxiliary substances
comprise additives, extenders, colorants, dyes, or flow
enhancers.
62. The method of claim 60, wherein the pancreatin is obtained from
the pancreas of a mammal.
63. The method of claim 60, further comprising i.) coating the
pellet or micropellet.
64. The method of claim 63, wherein the coating comprises
hypromellose phthalate, dimethicone 1000, and dibutyl phthalate,
monoglycerides, or an emulsifiable lipid.
Description
RELATED APPLICATIONS
[0001] This application is a divisional application of U.S. Utility
application Ser. No. 13/407,408, which is a continuation patent
application of U.S. Utility application Ser. No. 12/386,051, filed
Apr. 13, 2009, and is also related to international application no.
PCT/US10/30895, filed Apr. 13, 2010, which claims the benefit of
U.S. Utility application Ser. No. 12/386,051, each of which are
herein incorporated by reference in their entirety.
FIELD OF THE INVENTION
[0002] The present invention relates generally to coated
digestive/pancreatic enzyme preparations, and pharmaceutical
compositions and enzyme delivery systems comprising the
preparations, as well as methods for their preparation, use, and
controlled delivery in treating individuals with neurological or
behavioral diseases or conditions susceptible to treatment with
enzymes.
BACKGROUND
[0003] Digestive enzymes are produced by the salivary glands,
glands in the stomach, the pancreas, and glands in the small
intestines. For example, digestive enzymes produced by the pancreas
and secreted into the stomach and small intestine aid in digestion.
Digestive enzymes produced by the pancreas are secreted into the
duodenum, or upper segment of the small intestine, where the pH is
around 5 to 6, and the enzymes assist in the digestion of food
components, including carbohydrates, lipids, proteins and nucleic
acids. However, when digestive enzymes are administered orally, the
enzymes are exposed to highly acidic conditions in the stomach,
with a pH of around pH 1-2, as well as gastric proteases which
denature and degrade the enzymes.
[0004] Digestive enzymes have been administered to mammals to treat
enzyme deficiencies caused by conditions affecting the pancreas,
such as pancreatitis and pancreatic enzyme deficiency. Pancreatic
enzymes administered to humans are commonly of porcine origin.
Manufacturers of enzyme preparations have also used enteric
coatings for lipase compositions in individuals with cystic
fibrosis who require administration of lipases. The preparations
for lipase delivery have used enteric coatings containing, for
example, hypromellose phthalate, dimethicone 1000, and dibutyl
phthalate.
[0005] Certain methods for coating sensitive bioactive substances
have been described. U.S. Pat. No. 6,261,613 to Narayanaswamy et
al. discloses particles that can contain yeast, coated in a shell
of a fat in a beta prime form (i.e., triglyceride crystals having a
blocky symmetry). The coating material can further contain
emulsifiers such as those found in hydrogenated vegetable oil.
However, the coating only allows release of the yeast in a limited
temperature range of about 40.degree. C. to about 55.degree. C.
U.S. Pat. No. 6,251,478 B1 to Pacifico et al. discloses certain
sensitive substances including certain bioactive compounds
encapsulated in a lipid material.
[0006] No description in the Background section should be taken as
an admission that such disclosure constitutes prior art to the
instant invention.
SUMMARY OF THE INVENTION
[0007] The present invention relates to coated digestive enzyme
preparations, and pharmaceutical compositions and enzyme delivery
systems comprising coated digestive enzyme preparations, which are
useful in the treatment of individuals with autism, ADD, ADHD,
Parkinson's disease, cystic fibrosis, other neurological and
behavioral diseases or conditions. The coated and encapsulated
digestive enzyme preparations of this invention permit controlled
delivery of enzymes having increased stability and enhanced
administration properties, to patients with neurological and
behavioral diseases and conditions susceptible to treatment with
digestive enzymes.
[0008] In some aspects, the present invention relates to a coated
and/or encapsulated pancreatic/digestive enzyme preparation which
comprises a core comprising digestive and/or 20 pancreatic enzymes
and a coating which comprises an emulsifiable lipid. The core
contains an amount of pancreatic/digestive enzyme effective for
treatment of the patient's condition, which can be, for example, a
neurological disorder such as autism, ADD, ADHD, CF and Parkinson's
disease, or other diseases for which an effective amount of
pancreatic/digestive enzymes can be administered. Among other
properties, the coating protects the pancreatic/digestive enzyme
from destabilizing factors such as solvents, heat, light, moisture
and other environmental factors. The coating also provides
controlled release of the pancreatic/digestive when the composite
is exposed to a solvent. In addition, in one aspect of this
invention, the coated digestive enzyme preparations of this
invention have improved pour properties, and improved taste and
smell of the digestive enzyme particles.
[0009] The invention also relates to a specific blend of enzymes
and lipids for enzyme administration in individuals with
Parkinson's disease, ADD, ADHD, autism and cystic fibrosis and
other behavioral or neurological conditions and diseases. The
coated digestive enzyme preparations can be used to obtain release
at selected transit times or in selected locations of the
gastrointestinal tract of humans. In one aspect, this invention
relates to controlled release enzyme preparations.
[0010] In another aspect the invention relates to a coated
digestive enzyme preparation comprising (a) a core containing a
digestive enzyme particle, where the enzyme present in an amount of
from about 5% to 90% by weight of the particles; and (b) a coating
comprising an emulsifiable lipid, wherein the coating continuously
coats the core and the emulsifiable lipid emulsifies upon exposure
to a solvent.
[0011] In another aspect, this invention relates to a
pharmaceutical composition comprising a therapeutically effective
amount of an encapsulated enzyme preparation, which comprises (a) a
core which comprises an amount of pancreatic or digestive enzymes
effective for treating a subject suffering from autism, ADD, ADHD,
Parkinson's' disease, cystic fibrosis, or other neurological
condition or behavioral disorder susceptible to treatment by the
enzymes; and (b) a coating comprising an emulsifiable lipid.
[0012] In yet another aspect, this invention relates to an enzyme
delivery system comprising encapsulated enzyme preparation having
particles which comprise: (a) a core 20 comprising pancreatic or
digestive enzymes present in an amount of from about 5% to 95% by
weight of the particles; and (b) a generally uniform coating to
provide for controlled release of the enzymes, said coating
comprising an emulsifiable lipid. In one aspect, the encapsulated
enzyme preparation particles of the enzyme delivery system are
non-aerosolizable.
[0013] In certain aspects, the methods of preparing enzymes
according to this invention produce coated enzyme preparations
characterized, for example, by controlled rates of release,
reduction in aerosolization and safer administration, ability to be
administered by a sprinkle/sachet delivery method, improved flow
characteristics, enhanced shelf life and storage capacity, and
other properties described herein. In other aspects, the coated
enzyme preparation has improved pour properties which facilitate
manufacturing and packaging processes, for example packaging in
pouches and sachets.
[0014] In some aspects, the present invention is based on the
surprising and unexpected discovery that certain coated digestive
enzyme preparations which comprise a coating of emulsifiable lipid
and a digestive enzyme core have favorable release and activity
profiles and permit site time specific and/or location specific
targeted release along the GI tract for the treatment of autism,
ADD, ADHD, Parkinson's Disease and other neurological or behavioral
conditions susceptible to treatment with digestive enzymes. In some
aspects, the encapsulated pancreatic/digestive enzyme preparations
are prepared to obtain specific delivery times or specific regions
within the human gastrointestinal (GI) tract. In other aspects, the
emulsifiable lipid composition is hydrogenated soy oil, but may be
any suitable lipid or lipid blend.
[0015] The invention further relates in some aspects to more stable
enzyme preparations protected against the environment to reduce,
for example, degradation and/or denaturation of the enzymes. This
permits delivery of more accurate doses of the enzyme preparation
to treated individuals. The coating can also, in some aspects,
provide emulsification when the enzyme preparations are contacted
with appropriate solvents, while also surprisingly providing for
controlled release of the enzyme in the gastrointestinal (GI)
system. The emulsification properties of the coating in a solvent
allows for controlled release of the enzyme, preferably at selected
locations in the GI tract, where enzyme utilization provides the
most effective treatment.
[0016] The present invention also relates to methods of making the
enzyme preparations by lipid coating and/or encapsulation of
digestive enzymes. The methods comprise providing an emulsifiable
lipid, and coating screened pancreatic/digestive enzyme particles
with the lipid. The digestive enzymes comprise 5-95% of the coated
enzyme preparations by weight.
[0017] In another aspect, as described herein, the inventors have
surprisingly discovery that the methods of this invention can be
used to produce coated digestive enzyme preparations comprising
digestive and/or pancreatic enzymes coated with an emulsifiable
lipid alone, or with a lipid blend to achieve a controlled rate of
enzyme release, with increased release of the pancreatic/digestive
enzyme upon exposure of the encapsulated preparation to a suitable
solvent.
[0018] The inventors have discovered that encapsulated
pancreatic/digestive enzyme preparations having a coating
consisting essentially of one or more monoglycerides exhibit
increased release of the pancreatic/digestive enzyme upon exposure
of the encapsulated composite to a solvent, such as water, while
protecting against release in 0.1 N HC1.
[0019] The invention further relates to methods for administering
the enzyme preparations. In some aspects, the methods include
administering the pancreatic/digestive enzymes as coated
preparations. In some aspects, the invention relates to a method of
treatment comprising administering to a subject with autism, ADD,
ADHD, Parkinson's disease, cystic fibrosis, or other behavioral or
neurological condition in need of treatment with digestive enzymes,
at least two doses of a composition comprising a therapeutically
effective amount of an encapsulated digestive enzyme preparation
comprising a core comprising a digestive enzyme; and a coating
comprising an emulsifiable lipid. Determination of whether a
subject is in need of treatment with an effective amount of
digestive enzymes may be based on a determination that the subject
has an enzyme deficiency.
[0020] In addition, the invention relates to the delivery to humans
of pancreatic/digestive enzyme composites, preparations, enzyme
delivery compositions or systems comprising no or fewer excipients,
carriers, additives and/or extenders, and/or requiring the use of
no or fewer solvents' in the enzyme preparations. In some
embodiments, the coating consists essentially of hydrogenated soy
oil. This can reduce exposure to potentially toxic substances and
will also reduce the possibility of allergy formation. The
invention further relates to the delivery of pancreatic and/or
digestive enzymes with improved safety of administration.
[0021] In addition, the invention relates to methods of improved
manufacturing resulting from the enhanced flow properties imparted
to enzyme preparations by the lipid encapsulation. The lipid
encapsulation of pancreatic/digestive enzymes forms a lipid barrier
to moisture which permits improved flow of the encapsulated enzyme
preparations in the packaging machinery.
[0022] The summary of the invention is not intended to be a
complete or exhaustive recounting of every aspect of the invention
described herein. Other aspects of the invention will be apparent
from further description set forth herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0023] FIG. 1 shows an electron micrograph of an unprocessed, raw
digestive enzyme particles;
[0024] FIG. 2 shows an electron micrograph of a coated enzyme
preparation following sieving and lipid coating of the raw
digestive enzyme preparation;
[0025] FIG. 3 shows a bar graph particle size analysis for a raw
digestive enzyme particles with the % of particles that can pass
through a USSS sieve, as indicated on the y-axis;
[0026] FIG. 4 shows a bar graph of the % lipase activity in the raw
digestive enzyme particles, and following encapsulation, for coated
enzyme preparations containing 70%, 80% and 90% digestive enzymes
by weight;
[0027] FIG. 5 shows a bar graph of the % enzyme release for the
enzyme preparations containing 70%, 80% and 90% digestive enzymes
by weight, at the times indicated on the x-axis.
[0028] FIG. 6 shows a bar graph of the particle size distributions
of the raw digestive enzyme particles compared with the particle
size distributions in coated enzyme preparations containing 70% or
80% digestive enzymes by weight;
[0029] FIG. 7 shows the flow chart for a process that can be used
to encapsulate digestive enzyme particles;
[0030] FIG. 8 shows a chromatogram of peak area (mAU) vs. time for
working standard (top line), diluent (line that starts third from
the top when time is 4 minutes), mobile phase used in the HPLC
(bottom line at 4 minutes) and placebo (second to the top line when
time is 4 minutes), which demonstrate no interference with the
standard trypsin peak.
[0031] FIG. 9 shows a graph of peak area (mAU) vs. sample
concentration (mg/mL) for known trypsin concentrations obtained
using HPLC to measure trypsin in the coated digestive enzyme
preparation.
[0032] FIG. 10 shows fecal chymotrypsin (FCT) levels measured in
nine children with symptoms of autism.
[0033] FIG. 11 shows FCT levels measured in 26 children with
symptoms of autism.
[0034] FIG. 12 shows FCT levels measured in 46 children. 25 of the
children had symptoms of autism, while 21 children did not have
symptoms of autism.
[0035] FIG. 13 shows fecal chymotrypsin levels measured in 320
age-matched children. The navy line (in grayscale, the upper, black
line) shows FCT levels for children with known conditions (genetic
and other conditions). The purple line (in grayscale, the upper,
dark gray line), shows FCT levels for normal children without any
known condition. The aqua line, (in gray scale, the lower, medium
gray line), shows FCT levels for children with autism. The pink
line (in gray scale, the lower, dark gray line), shows FCT
measurements for children with ADHD. The yellow line (in grayscale,
the lower, light gray line), shows FCT measurements for children
with ADD.
[0036] FIG. 14 shows mean fecal chymotrypsin levels at baseline,
and 30, 60, 90 and 120 days after administration of Viokas or
Ultrase enzyme replacement.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0037] As described throughout, this invention relates in some
embodiments to coated digestive enzyme preparations, and
pharmaceutical compositions and enzyme delivery systems comprising
coated digestive enzyme preparations, which are useful in the
treatment of individuals with autism, ADD, ADHD, Parkinson's
disease, cystic fibrosis, other neurological and behavioral
diseases or conditions.
[0038] Autism (sometimes called "classical autism") is the most
common condition in a group of developmental disorders known as the
autism spectrum disorders (ASDs). Autism is characterized by
impaired social interaction, problems with verbal and nonverbal
communication, and unusual, repetitive, or severely limited
activities and interests. Other ASDs include Asperger syndrome,
Rett syndrome, childhood disintegrative disorder, and pervasive
developmental disorder not otherwise specified (usually referred to
as PDD-NOS). It has been estimated that three to six children out
of every 1,000 will have autism.
[0039] Attention deficit-hyperactivity disorder (ADHD) is a
neurobehavioral disorder that affects 3-5 percent of all children
in the US. It interferes with a person's ability to stay on a task
and to exercise age-appropriate inhibition (cognitive alone or both
cognitive and behavioral). Some of the warning signs of ADHD
include failure to listen to instructions, inability to organize
oneself and school work, fidgeting with hands and feet, talking too
much, leaving projects, chores and homework unfinished, and having
trouble paying attention to and responding to details. There are
several types of ADHD: a predominantly inattentive subtype, a
predominantly hyperactive-impulsive subtype, and a combined
subtype. ADHD is usually diagnosed in childhood, although the
condition can continue into the adult years.
[0040] Parkinson's disease (PD) belongs to a group of conditions
called motor system disorders, which are associated with the loss
of dopamine-producing brain cells. The four primary symptoms of PD
are tremor, or trembling in hands, arms, legs, jaw, and face;
rigidity, or stiffness of the limbs and trunk; bradykinesia, or
slowness of movement; and postural instability, or impaired balance
and coordination. As these symptoms become more pronounced,
patients may have difficulty walking, talking, or completing other
simple tasks. PD usually affects people over the age of 50. Early
symptoms of PD are subtle and occur gradually. In some people the
disease progresses more quickly than in others. As the disease
progresses, the shaking, or tremor, which affects the majority of
PD patients may begin to interfere with daily activities. Other
symptoms may include depression and other emotional changes;
difficulty in swallowing, chewing, and speaking; urinary problems
or constipation; skin problems; and sleep disruptions.
[0041] Cystic fibrosis (CF) is one of the most common
life-shortening, genetic diseases. In the United States, 1 in 4,000
children are born with CF. It is most common among western European
populations; one in twenty-two people of Mediterranean descent are
carriers of one gene for CF, making it the most common genetic
disease in these populations. CF is caused by a mutation in the
gene, cystic fibrosis transmembrane conductance regulator (CFTR).
The product of this gene is a chloride ion channel important in
creating sweat, digestive juices, and mucus. Although most people
without CF have two working copies (alleles) of the CFTR gene, only
one is needed to prevent cystic fibrosis. Cystic fibrosis affects
the exocrine (mucus) glands of the lungs, liver, pancreas, and
intestines, causing progressive disability due to multisystem
failure. CF can be characterized by, for example, 1) thick mucus
production which results in frequent lung infections; 2) diminished
secretion of pancreatic enzymes causing poor growth, greasy stools,
and deficiency in fat-soluble vitamins; and 3) infertility in the
males due to the condition congenital bilateral absence of the vas
deferens. Often, symptoms of CF appear in infancy and childhood.
Meconium ileus is a typical finding in newborn babies with CF.
[0042] Enzyme preparations with non-lipid enteric coatings have
been used to deliver lipases in individuals requiring
administration of lipases to individuals with cystic fibrosis in
need of enzyme treatment. In addition, Fallon has described certain
methods and enzyme compositions for use in treating children and
other individuals, with autism, ADD, ADHD, Parkinson's disease and
other neurological diseases or conditions, for example, U.S. Pat.
Nos. 7,138,123, 6,660,831, 6,632,429; 6,534,063, hereby
incorporated by reference as if set forth in full herein.
[0043] The nature of the human digestive tract creates challenges
for the delivery of digestive enzymes to patients with neurological
and behavioral conditions susceptible to treatment with digestive
enzymes. Multiple temperature and pH changes over the course of the
digestive tract make specific delivery a necessity and a challenge.
For instance, pH as low as 1 is encountered in the stomach, but
rapidly increases to a more basic pH of 5-6 in the proximal small
intestine. For example, generally the pH in the stomach is
approximately 1.2, the pH in the duodenum is about 5.0 to 6.0; the
pH in the jejunum is about 6.8, and the pH is about 7.2 in the
proximal ileum and about 7.5 in the distal ileum. The low pH in the
stomach which changes rapidly to a more basic pH of 5-6 in the
proximal small intestines, call for a specific delivery 25 method
depending upon where the enzyme is to be delivered.
[0044] For example, children with cystic fibrosis whose condition
requires administration of lipases, require delivery of the lipases
to the latter portion of the small intestine. In contrast, the
inventors have determined that children with autism who need
treatment with proteases require delivery of those enzymes to the
proximal small intestine.
[0045] Delivery of digestive enzymes can also be challenging due to
the rapid degradation and denaturing of enzymes at ambient room
temperature, as well as the enhanced degradation and denaturing
that can occur with high temperature, pressure, humidity and/or
exposure to light. Moisture and heat together can quickly
destabilize enzymes, reducing their effectiveness, and shortening
shelf life, leading to inaccurate dosing. Denaturization or
destabilization of the enzymes can reduce their effectiveness by
reducing the dose of active enzymes to less than the amount needed
for effective treatment. Alternatively, attempting to compensate
for the denaturization or destablization by increasing the dose to
ensure an effective level of active enzyme, could risk an overdose
or overfilling a capsule or other dosage form. To protect and
stabilize the pancreatic/digestive enzyme from unfavorable
conditions, such a penetration, decomposition, the
pancreatic/digestive enzyme (core) may be coated or encapsulated in
a continuous coating containing an emulsifiable lipid. In another
aspect, this invention provides new coated enzyme preparations with
improved shelf life.
[0046] Manufacturers of enzyme preparations have used enteric
coatings to deliver lipases in individuals requiring administration
of lipases, such as individuals with cystic fibrosis. Because the
porcine enzymes are delivered in a mixture of proteases, lipases
and amylases, and because these compositions for human consumption
were prepared for lipase delivery, the use of these enteric
coatings, which include such substances as hypromellose phthalate,
dimethicone 1000, and dibutyl phthalate, preclude delivery of
proteases at the proper location in the digestive tract. All other
enzyme preparations presently on the market contain at least one of
these enteric coating substances and/or other additives in the
preparation. Some additives that enable manufacturing, such as
additives to improve flow properties, may further risk patient
reactivity or sensitivity to the enzyme preparation.
[0047] In one embodiment the present invention includes a coated
digestive enzyme preparation and/or composite, which, in some
embodiments is an encapsulated pancreatic/digestive enzyme
preparation. In other aspects, the invention includes enzyme
delivery systems and pharmaceutical compositions comprising coated
pancreatic/digestive enzyme preparations. These coated or
encapsulated enzyme preparations contain cores comprising
pancreatic or digestive enzyme particles, and a coating comprising
an emulsifiable lipid.
[0048] The coatings in the digestive/pancreatic enzyme preparations
create a barrier to degradation and denaturation, and allow more
accurate levels of active enzymes to reach the treated individuals.
The lipid coating of this invention provides a significant barrier
to moisture, heat, humidity and exposure to light by allowing for a
physical barrier as well as one that prevents and or reduces
hydrolysis. The coated enzyme preparations undergo less hydrolysis
as a result of protection from moisture in the environment by the
lipid coating. As a result of the present invention,
pancreatic/digestive enzymes are provided which can tolerate
storage conditions (e.g., moisture, heat, oxygen, etc.) for long
periods of time thus enabling extended shelf life. The coating of
the encapsulated enzyme preparation protects the enzyme from the
environment and provides emulsification in a solvent without
detracting from the abrasion resistance of the coating. The
invention thus further relates to more stable enzyme
preparations.
[0049] The coated enzyme preparations therefore reduce overfilling
of the enzyme dosage, and enhance delivery of more accurate doses
of the enzyme to individuals with autism, ADD, ADHD Parkinson's
disease, cystic fibrosis and other neurological or behavioral
conditions or diseases susceptible to treatment with pancreatic or
digestive enzymes.
[0050] In addition, because children and other individuals with
autism and other conditions often have multiple sensitivities to
foods, additives, colorants and other carriers, excipients or
substances used in drug formulations, it is a challenge to make an
enzyme delivery system that avoids the use of allergens, and other
carriers, excipients, extenders, colorants, etc. that could
potentially add to adverse symptoms or the morbidity of patients.
Furthermore, in very young children an enzyme delivery system which
allows ease and tolerability, is paramount. A sachet delivery
system for these enzyme preparations has also heretofore not been
achieved.
[0051] It is another aspect of the present invention to make an
enzyme preparation without the use of extenders colorants, dyes,
flow enhancers and other additives to reduce the potential for
allergens and other sensitivity reactions in children and other
treated individuals. It has been discovered that in some
embodiments, the digestive enzymes can surprisingly be encapsulated
with a single lipid excipient to improve retention of enzyme
activity, ease of administration, tolerability, and safety of
administration, among other properties. Surprisingly digestive
enzyme particles containing lipases can be successfully
encapsulated with coating consisting essentially of only
hydrogenated soy oil.
[0052] In addition, porcine pancreatic/digestive enzymes posses a
significant odor and taste, similar to cured/smoked pork. This
taste can be strong and offensive to some individuals taking enzyme
replacement, and especially to children. The addition of a lipid
coating provides significant taste masking to the enzyme
preparation, which allows for the tolerance of taste, as the lipid
coating is odorless and tasteless. The use of this method of taste
masking which does not involve the use of color, dyes, perfumes,
recipients, or other substances is preferable for the
administration of medications, which have an unpleasant or
undesirable taste and odor. In other embodiments, this invention
relates to coated digestive enzyme preparations with improved taste
and smell.
[0053] In some embodiments, the coatings on the digestive enzyme
particle cores are preferably continuous coatings. By "continuous,"
it is meant that the pancreatic/digestive enzyme is uniformity
protected. The continuous coating of the fully surrounds or
encapsulates the pancreatic/digestive enzymes. The encapsulation
provides protection of the pancreatic/digestive enzyme from
conditions such as moisture, temperature, and conditions
encountered during storage.
[0054] In addition, the encapsulation also provides controlled
release of the pancreatic/digestive enzyme. The emulsification
properties of the coating in a solvent allows for controlled
release of the enzyme in the gastrointestinal system, preferably
the region of the GI tract where the enzymes are to be utilized.
The coating of the encapsulated composite protects the enzyme from
the environment and provides emulsification in a solvent without
detracting from the abrasion resistance of the coating. For
example, for conditions requiring treatment with proteases, the
release of the protease portion of the enzymes is necessary in the
proximal small intestine, thereby necessitating a lipid
encapsulation which has a dissolution profile between 30-90
minutes. The dissolution profile may also be about 30, 35, 40, 45,
50, 55, 60, 65, 70, 75, 80, 85 or 90 minutes. Dissolution profiles
may be obtained using methods and conditions known to those of
skill in the art. For example, dissolution profiles can be
determined at various pH's, including pH. 1, 2, 3, 4, 5, 6, 7, 8,
9, 10.
[0055] The rate of release of the bioactive substance can also be
controlled by the addition of additives as described below. When
the preparations are exposed to a solvent, the solvent interacts
with the mollifiable lipid in the coating and results in
emulsification of the coating and release of the bioactive
substance.
[0056] "Encapsulate" as used herein means that the coating
completely surrounds the pancreatic/digestive enzyme. In a
population of encapsulated particles, encapsulated enzyme
preparations may include contaminating or small portion of
particles with a substantially continuous coating as long as the
release profiles of the encapsulated particles are not
significantly altered. A coated or encapsulated particle may
contain one or more digestive enzyme particles enveloped in one
coating to form one coated or encapsulated digestive enzyme
particle in the coated or encapsulated digestive enzyme
preparation.
[0057] The present invention also includes a method for preparing
the enzyme preparations, pharmaceutical compositions, and delivery
systems for the treatment of neurological or behavioral disorders
such as autism, ADD, ADHD Parkinson's disease, cystic fibrosis and
other behavioral or neurological conditions or diseases susceptible
to treatment with pancreatic or digestive enzymes. By "susceptible
to treatment with pancreatic or digestive enzymes" is meant that
one or more symptoms of the disease or condition can be alleviated,
treated, or reduced by administration of an effective amount of
pancreatic or digestive enzymes.
[0058] In some aspects, the invention relates to the production of
selected coated enzyme preparations made by coating digestive
enzyme particles with lipids not previously used in coated
digestive enzyme preparations. The unique mixtures of emulsifiable
lipids and enyzmes can deliver certain components of the
pancreatic/digestive enzymes to selected locations and/or at
selected times during transit of the GI tract. In some aspects, the
invention relates to methods of delivering digestive enzymes to
humans based upon dissolution profiles.
[0059] The emulsifiable lipid is any lipid, lipid mixture, or blend
of lipid and emulsifiers which emulsifies when exposed to a
solvent, and has a melting point which allows the lipid to be a
solid at typical storage temperatures. The emulsifiable lipid can
be a vegetable or animal derived-lipid. In some embodiments, the
emulsifiable lipid consists essentially of, or comprises one or
more monoglycerides, diglycerides or triglycerides, or other
components including, for example, emulsifiers found in
hydrogenated vegetable oils. In another embodiment the lipid is a
non-polar lipid.
[0060] As used herein, animal and/or vegetable "derived" lipids can
include fats and oils originating from plant or animal sources
and/or tissues, and/or synthetically produced based on the
structures of fats and oils originating from plant or animal
sources. Lipid material may be refined, extracted or purified by
known chemical or mechanical processes. Certain fatty acids present
in lipids, termed essential fatty acids, must be present in the
mammalian diet. The lipid may, in some embodiments, comprise a Type
I USP-National Formulary vegetable oil.
[0061] The digestive enzyme used in the present invention can be
any combination of digestive enzymes of a type produced by the
pancreas, including, but not limited to digestive enzymes from a
pancreatic source or other sources. The scope of the invention is
not limited to pancreatic enzymes of porcine origin, but can be of
other animal or plant origin as well as those which are
synthetically derived. The digestive enzyme may be derived from
mammalian sources such as porcine-derived digestive enzymes. The
enzyme may include one or more enzymes, and can also be plant
derived, synthetically derived, recombinantly produced in
microbial, yeast, or mammalian cells, and can include a mixture of
enzymes from one or more sources. Digestive enzyme, can include,
for example, one or more enzymes from more or more sources mixed
together. This includes, for example, the addition of single
digestive enzymes to digestive enzymes derived from pancreatic
sources in order to provide appropriate levels of specific enzymes
that provide more effective treatment for a selected disease or
condition. One source of digestive enzymes can be obtained, for
example, from Scientific Protein Laboratories (see Table 6). The
digestive enzyme may be, for example a pancreatin/pancrelipase
composition. In one embodiment, the digestive enzymes will comprise
or consist essentially of 25 USP units/mg protease, 2 USP Unit/mg,
and 25 USP Units/mg amylase. The term digestive enzyme may refer to
one or more enzymes of a type produced by the pancreas.
[0062] The digestive enzyme particles used as cores in the present
invention include digestive enzyme particles where about 90% of the
particles are between about #40 and #140 USSS mesh in size, or
between about 105 to 425 .mu.m, or where at least about 75% of the
particles are between about #40 and #80 mesh, or about 180 to 425
.mu.m in size. Particles between #40 and #140 mesh in size pass
through #40 mesh but do not pass through #140 mesh. The coated or
encapsulated digestive enzyme particles in one embodiment of this
invention may comprise less than about 35, 30, 25, 20, 15 or 10% of
the particles which can be sieved through #100 mesh (150 .mu.m). In
some embodiments, the term "non-aerosolizable" refers to a coated
or encapsulated enzyme preparation where less than about 20% or
less than about 15% of the particles can be sieved through #100
mesh (150 .mu.m). The encapsulated digestive enzyme preparation can
be an encapsulated digestive enzyme composite where the digestive
enzyme particles contain two or more enzymes.
[0063] The minimum amount of pancreatic enzyme present in the core
is at least about 5% active enzymes by weight of the coated enzyme
preparation, but in other embodiments may be at least about 30%, or
at least about 50% by weight. The maximum amount of
pancreatic/digestive enzyme present in the composite is at most
about 95% by weight, and in other embodiments at most about 90%,
85%, 80%, 75% or 70% of the coated enzyme preparation. In other
embodiments, the amount of pancreatic enzyme present in the
composite is about 10%, 15%, 20%, 25%, 35%, 40%, 45%, 55%, 60%,
65%, 70%, 72.5%, 75%, 77.5%, 80%, 82.5%, 87.5%, or 92.5% by weight
or anywhere in between. At least about or at most about a of enzyme
may include equal to or about that % of enzyme. The term "about"
includes equal to, and a range that takes into account experimental
error in a given measurement. As used in connection with particle
sizes, the term "about" can refer to plus or minus 10, 9, 8, 7, 6,
5, 4, 3, 2 or 1% or anywhere inbetween. As used in connection with
% particles that can be sieved, the term "about" can refer to plus
or minus 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1% or anywhere
inbetween.
[0064] The composition which contains the encapsulated digestive
enzyme preparation or composite can be delivered as a sprinkle,
powder, capsule, tablet, pellet, caplet or other form. Packaging
the encapsulated enzyme preparations in an enzyme delivery system
that further comprises single dose sachet-housed sprinkle
preparations allows for ease of delivery, and accurate dosing of
the enzyme, by allowing a specific amount of enzyme to be delivered
in each dosing. Allowing for specific unit dosing of an enzyme
preparation which maintains the enzyme activity within specific
stability parameters in an enhancement over other sprinkle
formulations, which are housed, in a multi-unit dosing form that
allows for air, moisture and heat to depredate and denature the
enzyme preparation. In a preferred embodiment the powder or sachet
is housed in a trilaminar foil pouch, or similar barrier to keep
out moisture and to protect the enzyme preparation from adverse
environmental factors. The invention further relates to an
improvement in stability due to a reduction in hydrolysis due to
the lipid encapsulation.
[0065] Further the lipid encapsulation methodology reduces the
aerosolization of the enzyme preparation that may be caustic to the
child if inhaled through the lungs or the nose. In another
embodiment, the invention includes delivery of digestive enzymes
with improved safety of administration, by reducing the amount of
aerosolization of the enzyme. The lipid encapsulation reduces
aerolization and the potential for caustic burn, aspiration, and/or
aspiration pneumonias in children and administrators of the enzyme
preparation, thereby reducing the potential for illness in already
compromised children such as those with cystic fibrosis, and
leading to safer administration.
[0066] As used herein, the term "non-aerosolizable" will be used to
refer to a coated or encapsulated enzyme preparation where
substantially all of the particles are large enough to eliminate or
reduce aerosolization upon pouring of the coated enzyme preparation
compared to uncoated enzyme particles. For example, the term
"non-aerosolizable" may refer to a coated or encapsulated enzyme
preparation where at least about 90% of the particles are between
about #40 and #140 mesh in size, or between about 106 to 425 .mu.m,
or where at least about 75% of the particles are between about #40
and #80 mesh, or about 180 to 425 .mu.m. The term "non
aerosolizable" may also refer to a coated or encapsulated enzyme
preparation where less than about 35, 30, 25, 20, 15 or 10% of the
particles can be sieved through #100 mesh (150 .mu.m). In some
embodiments, the term "non-aerosolizable" refers to a coated or
encapsulated enzyme preparation where less than about 20% or less
than about 15% of the particles can be sieved through #100 mesh
(150 .mu.m).
[0067] As described and referred to herein, suitable
pancreatic/digestive enzymes and suitable coatings may be used in
the compositions and methods of this invention. The choice of
suitable enzymes and of suitable lipid coatings, including choice
of the type or amount of enzymes or coating, are guided by the
specific enzyme needs of the individuals, and the selected diseases
to be treated. The encapsulated enzyme preparations that are one
aspect of this invention have not been previously described.
[0068] In some embodiments, the invention relates to specific
blends of enzymes and lipids selected for delivery in individuals
with Parkinson's disease, ADD, ADHD, autism, cystic fibrosis and
other neurological and behavioral disorders susceptible to
treatment with digestive/pancreatic enzymes based on the transit
times in the human gastrointestinal tract. It can further be based
upon the need of the patient to be treated for various components
of the digestive enzymes. Further, the invention relates to
improvement of the delivery of digestive enzymes to humans based
specifically upon required delivery times, and dissolution
profiles.
[0069] While general methods for coating certain sensitive biologic
substances have been described, see, e.g., U.S. Pat. No. 6,251,478,
hereby incorporated by reference, the encapsulated bioactive
substance of this invention is an enzyme preparation comprising a
core containing digestive enzymes comprising or consisting of
multiple proteases, lipases and amylases, and a coating which
comprises or consists essentially of an emulsifiable lipid.
[0070] Additives can be blended with the emulsifiable lipid.
Selection of the lipid(s) and additives will control the rate of
release of the bioactive substance. In the case of the digestive
and or pancreatic enzymes, the lipid coat must be uniquely chosen
to release the bioactive substance in the area of the digestive
tract selected for release to optimize treatment.
[0071] The invention further relates to the administering of the
coated and/or encapsulated enzyme preparation in a sachet or pouch
preparation for ease of delivery to children and adults. In some
embodiments, the invention specifically relates to the
administration of a coated enzyme particle preparation, housed in a
sachet or pouch. This facilitates administration, including but not
limited to, administration in food or drink, direct administration
into the oral cavity, or administration directly into the GI system
through an NG-tube, G-tube or other GI entrances or deliveries.
[0072] In some embodiments, each dose contains about 100 to 1500 mg
of coated or encapsulated enzyme preparation, and each dose may
contain about 100, 150, 200, 250, 300, 350, 400, 450, 500, 550,
600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150,
1200, 1250, 1300, 1350, 1400, 1450, or 1500 mg of coated or
encapsulated enzyme preparation. "About" can include 80 to 125% of
the recited preparation. Each dose may also be plus or minus 10% of
the recited weight. In one embodiment each does will have a
protease activity of not less than about 156 USP units/mg plus or
minus 10%. The protease activity may also be not less than about
100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160,
165, 170, 175, 180, 185, 190, 195, or 200 USP units/mg.
[0073] In other embodiments, the invention relates to methods of
treatment comprising administering to a subject with autism, ADD,
ADHD, Parkinson's' disease, cystic fibrosis, or other behavioral or
neurological condition susceptible to treatment with digestive
enzymes, at least two doses of a composition comprising a
therapeutically effective amount of the coated digestive enzyme
preparations. In certain embodiments, about 80% of the enzyme is
released by about 30 minutes in a dissolution test performed at pH
6.0. In other embodiments, about 80% of the enzyme is released by
about 30 minutes after the coated digestive enzyme preparations
reach the small intestine.
[0074] Another embodiment of the invention relates to the
improvement of delivery of enzymes to humans by reducing the use of
excipients, extenders and solvents currently used in the
preparations for delivery of digestive enzymes to humans. For
example, the encapsulated digestive enzyme preparation may contain
only one excipient, which increases the safety of administration by
decreasing the chance of an allergic response. In one embodiment,
the excipient is hydrogenated soy oil.
[0075] Because, in some embodiments, the lipid encapsulation method
does not require the enzyme preparation to be treated with
solvents, extenders and excipients to facilitate flow or improve
stability, one aspect of the invention includes a "clean"
preparation of GRAS substances (generally regarded as safe) to be
administered. The reduction in the use of solvents, extenders
excipients and other additives permitted by the methods of this
invention reduces the exposure of the individuals taking the enzyme
replacement to potential allergens, thereby producing a
hypoallergenic enzyme preparation that further enhances its
potential uses in the treatment of individuals who might otherwise
develop an allergic response to treatment. Administration of the
coated enzyme preparations of this invention can thus reduce
exposure to potentially toxic substances and will also reduce the
possibility of allergy formation. Accordingly, in some embodiments,
the encapsulated digestive enzyme preparation is
hypoallergenic.
[0076] The invention further relates in another aspect to the
delivery of digestive enzymes with improved safety of
administration. The lipid coat adds weight to the enzyme
preparation, which reduces the potential for aerosolization.
Previous uncoated enzymes have been shown to become aerosolized,
and can therefore be inhaled and contact the nasal cavity or the
lungs, causing injury to the mucosa of those taking and those
administering the enzyme preparation.
[0077] The invention further relates to the improvement of
administering a sachet preparation for delivery to children. The
invention specifically relates to the administration of a coated
digestive enzyme preparation, housed in a sachet which allows for
particular types of administration including but not limited to
administration in food, drink, or direct administration into the
oral cavity or directly into the GI system through a NG-tube,
G-tube or other GI entrances. The use of a sachet delivery of
enzymes has heretofore been not utilized in the enzyme preparations
presently marketed. The sachet, which represents a unit dosage or
multiple doses for a day, and represents a single unit dose. The
sachet of a trilaminar foil allows the enzyme/lipid powder to
remain stable, and allows for ease of administration.
[0078] In another embodiment, the invention relates to a method of
controlling the rate of release of the pancreatic/digestive enzyme
from an encapsulated enzyme preparation upon exposure to a solvent.
In some aspects, the method comprises blending an emulsifiable
lipid with an amount of one or more additives to obtain a lipid
blend; and coating the digestive enzyme particle with the blend to
form an encapsulated digestive enzyme preparation containing
particles comprising a core which contains the enzyme, and a
coating which contains the lipid. In some embodiments, the
emulsifiable lipid is a blend where the emulsifiable lipid and
additive are not the same, and where the rate of release of the
enzyme from the encapsulated composite upon exposure to a solvent
is decreased as the amount of additive is increased. In the
alternative, the rate of release of the enzyme from the
encapsulated composite upon exposure to a solvent is increased as
the amount of additive is decreased.
[0079] The lipid coating surprisingly does not appear to be reduced
or destroyed by HC1 (hydrochloric acid) present in the stomach,
thereby protecting the enzyme from degradation following
administration until the enzyme preparation reaches its target
region in the GI tract. Further the lipid coat reduces the exposure
of the enzyme to attack by water, thereby reducing hydrolysis, and
further protecting the digestive enzymes from degradation. In
addition, the inventors have found that an excipient containing
only lipid can be used to coat or encapsulate digestive enzyme
particles containing lipase.
[0080] The use of digestive enzymes for the treatment of specific
disease targets is made possible, in one aspect of the invention,
by preparing encapsulated digestive enzyme composite having
differing release characteristics. Since various neurological and
behavioral diseases can impact the gastrointestinal systems in
humans in various ways, the use of specific enzyme preparations and
the ensuing encapsulation can make the difference as to where and
for what duration of time the enzyme preparation is delivered.
[0081] The invention therefore relates to improvement of the
delivery of digestive enzymes to humans based specifically upon
needed delivery times, and dissolution profiles. For example, in
certain aspects of the invention, the rate of release and
dissolution characteristics are unique to the lipid encapsulations
of this invention. The preparation of coated digestive enzymes
using enzymes and lipids selected to optimize treatment of
behavioral and neurological conditions and diseases susceptible to
treatment with digestive enzymes has not been previously
described.
[0082] As an example, previous enteric coatings for digestive and
or pancreatic enzymes have delayed release of enzyme mixture for a
period of time too long for delivery of the protease portion to the
proximal small intestine. For instance, in administration to
patients with cystic fibrosis where delivery of lipases is required
for effective treatment, the dissolution profile of the enterically
coated digestive enzymes needs to favor a longer delay in the
release of the enzymes, as well as the delivery of a high lipase
formulation.
[0083] Prior to the instant invention, lipid encapsulation had not
been used as a delayed and/or protective mechanism for lipase
delivery to treat individuals with cystic fibrosis.
[0084] The inventors have further recognized that for treatment of
patients with autism who require delivery of protease enzymes for
effective treatment, the lipid encapsulate can be modified to
deliver the protease during an earlier transit time window, in the
proximal small intestine, to optimize protein digestion. In another
example, the inventors have recognized that for patients with
Parkinson's disease who have slow GI transit times due to the
dysautonomic nature of their neurological condition, still another
release profile is required to deliver enzymes for effective
treatment. The lipid and/or additive selection will be made to
obtain enzyme release at later times after administration.
[0085] It has not been previously appreciated that transit times
for digestive enzymes through the digestive system could be
controlled by layering lipids, or through encapsulation with
specific lipid types. In still another aspect, this invention
relates to a selected blend of enzymes and lipids for delivery in
individuals with Parkinson's disease, ADD, ADHD autism and cystic
fibrosis and other behavioral or neurological diseases or
conditions susceptible to treatment with pancreatic/digestive
enzymes, based upon the transit times in the gastrointestinal
systems of humans.
[0086] The invention further relates to an improvement in
manufacturing due to the enhanced flow properties imparted by the
lipid encapsulation. The improvement in manufacturing can also
accomplished through the lipid encapsulation of a
pancreatic/digestive enzyme due to the lipid barrier to moisture
thus allowing for improved flow in the packaging machinery. The
improved flow qualities may facilitate packaging of the coated
digestive enzyme preparations into, for example, pouches or
sachets.
[0087] In one aspect, this invention relates to the use of a lipid
encapsulation method to make a coated digestive enzyme preparation
for specific delivery times within the human gastrointestinal (GI)
tract targeted for use in the treatment of a specific disease or
condition. This disease or condition may be caused by or
characterized by a digestive deficit that can be treated by the
administration of digestive enzymes to the appropriate region of
the GI tract. The neurological or behavioral disease or condition
is one not traditionally associated with the digestive system,
where one or more symptoms can be treated by administering an
effective amount of a pancreatic and/or digestive enzyme
preparation.
[0088] Thus, the present specification is directed at lipid
encapsulation of specific enzymes targeted for use in the treatment
of specific diseases, and the encapsulation method includes the
amount and type of lipids used in the methods of this invention for
the preparation of the encapsulated digestive enzyme composite. The
present invention also relates to methods of making the enzyme
preparations by lipid coating and/or encapsulation of pancreatic
and/or digestive enzymes. The methods comprise providing an
emulsifiable lipid, and coating pancreatic/digestive enzyme
particles with the lipid, where the pancreatic/digestive enzymes
comprise 5-90% of the coated enzyme preparations by weight. In some
aspects the uncoated pancreatic/digestive enzyme particles have a
size range of about 105-425 .mu.m.
[0089] In one embodiment, the invention relates to a method of
preparing an encapsulated digestive enzyme preparation, the method
comprising a) screening uncoated digestive enzyme particles to
obtain particles of a suitable size for encapsulation; and b)
coating the screened digestive enzyme particles with an
emulsifiable lipid to form coated or encapsulated digestive enzymes
containing a core which contains the pancreatic/digestive enzyme
and a coating which contains the emulsifiable lipid. In some
embodiments, the encapsulated digestive enzyme preparation is a
controlled release digestive enzyme preparation, which may have
enhanced flow properties. The preparations may be useful in the
treatment of individuals with autism, ADD, ADHD, Parkinson's'
Disease, Cystic fibrosis and other neurological conditions.
[0090] Screening of the particles may include quality control steps
to improve the activity, appearance or particle size of the
digestive enzyme. For example, the particles may be analyzed to
determine enzyme activity content, and/or visualized using
chromatographic, microscopic or other analytical methods. The
particles may also be screened to obtain particles of a suitable
size for encapsulation by removing particles that are too fine or
too large. For example, the particles may be sieved to obtain
particles of a suitable size or more uniform size range for
encapsulation. As a further example, the particles may be sieved
through USSS #40 mesh and through USSS #140 mesh. Particles that
pass through the #40 mesh but are retained by the #140 mesh are of
an appropriate size range for coating or encapsulation Particles
may also be screened by sieving through USSS #140, #120, #100, #80,
#70, #60, #50, #45, or #40 mesh, or any combination thereof.
[0091] Enzyme preparations supplied by the API supplier may be
provided as irregular shaped, and multi-sized particles, with
uneven edges, and much clumping, and containing some crystalline
salt particles. (See, for example, FIG. 1). Uneven particle size
and shape reduces flow properties, and interferes with packaging.
In addition, pouring uncoated enzyme into the mouth of an
individual would be difficult, and potentially may cause too much
or too little of the enzyme to be delivered. Processing the
digestive enzyme particles according to methods in accordance with
one aspect of this invention yields a non-dusty, free-flowing
particulate preparation suitable for sachet packaging and for
pouring onto food or drink. In addition, as discussed throughout,
the use of lipid encapsulation to prevent aerosolization, and
therefore increase safety, to increase flow properties which
enhance manufacturing of a pharmaceutical is an embodiment of the
instant invention.
[0092] The size distribution of particles in an exemplary raw
enzyme preparation is shown in the graph in FIG. 3. Large particles
(>40 mesh) and very small particles (<140 mesh) are generally
not suitable for proper encapsulation and can be removed by
screening. In order to increase the flow properties of the
encapsulated pancreatic enzyme preparation, digestive enzyme
particles can be sieved to remove fines and overly large particles,
for example by including only particles of sizes 40-140 mesh, or
about 105 to 425 microns. In some embodiments, the coated digestive
enzyme preparation containing 80% digestive enzyme by weight is
made by coating sieved pancreatic enzyme particles with a
hydrogenated vegetable oil using 20 lbs. of enzyme particles and 5
lbs of hydrogenated vegetable oil.
[0093] In some embodiments, the temperature of the lipid or lipid
blend is maintained at 110.degree. F. before application to the
digestive enzymes, which are not heated.
[0094] In some embodiments, the lipid should be present in the
preparation at a minimum amount of about 5% by weight of the
encapsulated composite, preferably about 30%, and more preferably
about 50% by weight of the encapsulated composite. The maximum
amount of pancreatic/digestive enzyme present in the encapsulated
composite is about 95% by weight of the composite, preferably about
90%, and more preferably about 85% of the encapsulated composite.
The emulsifiable lipid can be any lipid or lipid-derived material
that emulsifies or creates an emulsion yet has a melting point
which allows the emulsifiable lipid to be a solid at typical
storage temperatures, for example, 23 degrees Centigrade.
[0095] "Emulsifiable lipids" as used herein means those lipids
which contain at least one hydrophilic group and at least one
hydrophobic group, and have a structure capable of forming a
hydrophilic and hydrophobic interface. These chemical and/or
physical properties, mentioned above, of an emulsifiable lipid
permit emulsification. Examples of interfaces include, for example,
micelles and bilayers. The hydrophilic group can be a polar group
and can be charged or uncharged.
[0096] The emulsifiable lipid can be derived from animal or
vegetable origins, such as, for example, palm kernel oil, soybean
oil, cottonseed oil, canola oil, and poultry fat, including
hydrogenated type I vegetable oils. In some embodiments, the lipid
is hydrogenated. The lipid can also be saturated or partially
saturated. Examples of emulsifiable lipids include, but are not
limited to, monoglycerides, diglycerides, fatty acids, esters of
fatty acids, phospholipids, salts thereof, and combinations
thereof.
[0097] The emulsifiable lipid is preferably a food grade
emulsifiable lipid. Some examples of food grade emulsifiable lipids
include sorbitan monostearates, sorbitan tristearates, calcium
stearoyl lactylates, and calcium stearoyl lactylates. Examples of
food grade fatty acid esters which are emulsifiable lipids include
acetic acid esters of mono- and diglycerides, citric acid esters of
mono- and di-glycerides, lactic acid esters of mono- and
di-gylcerides, polyglycerol esters of fatty acids, propylene glycol
esters of fatty acids, and diacetyl tartaric acid esters of mono-
and diglycerides. Lipids can include, for example, hydrogenated soy
oil.
[0098] Any emulsifiable lipid may be used in the methods and
products of this invention. In certain embodiments the emulsifiable
lipid used will produce non-agglomerating, non-aerosolizing enzyme
preparation particles.
[0099] In other embodiments, the method relates to preparation of
an encapsulated, controlled release digestive enzyme preparation
with enhanced flow properties useful in the treatment of
individuals with autism, ADD, ADHD, Parkinson's' Disease, Cystic
fibrosis and other neurological conditions, the method comprising:
a) blending an emulsifiable lipid with one or more additives to
obtain a blend; and b) coating screened digestive enzyme with the
blend to form an encapsulated digestive enzyme containing a core
which contains the digestive enzyme and a coating which contains
the blend of emulsifiable lipid.
[0100] The coating of the enzyme with the lipid, as shown in FIG.
2, allows for the enzyme to become more uniform in size and shape,
but reduces the jagged edges associated with the raw enzyme, and
allows for ease of administration and ease of manufacturing, as the
flow properties associated with the covered enzyme will allow for
the manufacturing machinery to easily fill the sachet/pouch with
the enzyme and reduces overfilling or under filing of the sachet.
The unit dose packaging reduces the ability of the child to open
the multi dose can/box/or other container. The trilaminar foil
pouch or sachet further reduces the ability of a child to open the
sachet/pouch, and over utilize the enzyme.
[0101] In another embodiment, the invention relates to a method of
controlling the rate of release of a digestive enzyme from the
encapsulated preparation by using a lipid blend to coat the
digestive enzyme. The method includes blending an emulsifiable
lipid with one or more additives to obtain a blend, and coating the
digestive enzyme with the blend to form an encapsulated digestive
enzyme containing a core which contains the digestive enzyme and a
coating which contains the blend of emulsifiable lipid. The rate of
release of the enzyme from the encapsulated preparation upon
exposure with a solvent is decreased as the amount of additive is
increased. In the alternative, the rate of release of the enzyme
from the encapsulated composite upon exposure with a solvent is
increased as the amount of additive is decreased. Thus, the nature
of the coating allows for controlled release of the enzyme from the
encapsulate.
[0102] Non-emulsifiable lipids do not possess the chemical and/or
physical properties related to emulsification as described above
and include any lipid, lipid derived material, waxes, organic
esters, or combinations thereof. Non-emulsifiable lipids generally
do not emulsify by themselves. Non-emulsifiable lipids can be used
as additives so long as the properties of the coating, and
constituent lipids, permit emulsification. Non-emulsifiable lipids,
such as, for example, triglycerides, can be blended with an
emulsifiable lipid of the present invention. The non-emulsifiable
lipid can be derived from animals, vegetables, mineral, or
synthetic origins. The non-emulsifiable lipid is preferably
hydrogenated, and can be saturated or partially saturated, and
includes, but is not limited to triglycerides. In a preferred
embodiment, the coating contains a blend of monoglycerides and
triglycerides applied to a pancreatic/digestive enzyme.
[0103] The inclusion of one or more additives with an emulsifiable
lipid of the present invention is used to control emulsification of
the coating and release of the enzyme. For example, the additive,
triglyceride, can be blended with monoglycerides (e.g., an
emulsifiable lipid), to control emulsification of the coating and
thus control (e.g., decrease) the rate of release of the enzyme
from the composite. As a further example, one or more additives,
such as a diglyceride and a triglyceride can be blended with the
emulsifiable lipid to control the rate of release of the enzyme.
Hydrogenated vegetable oils may contain emulsifying agents, such as
soy lecithin or other components.
[0104] Properties including mechanical strength, melting point, and
hydrophobicity can be considered when choosing a suitable lipid
coating for the digestive enzyme. Lipids having lower melting
points or more polar, hydrophilic properties were generally less
suitable for encapsulation because they resulted in product that
would cake under accelerated storage stability conditions. Enzyme
preparations made using, for example, hydrogenated soy oil,
hydrogenated castor wax, and carnauba wax all demonstrated good
pouring and no caking.
[0105] The wax can be paraffin wax; a petroleum wax; a mineral wax
such as ozokerite, ceresin, or montan wax; a vegetable wax such as,
for example, camuba wax, bayberry wax or flax wax; an animal wax
such as, for example, spermaceti; or an insect wax such as
beeswax.
[0106] Additionally, the wax material can be an ester of a fatty
acid having 12 to 31 carbon atoms and a fatty alcohol having 12 to
31 carbon atoms, the ester having from a carbon atom content of
from 24 to 62, or a mixture thereof. Examples include myricyl
palmitate, cetyl palmitate, myricyl cerotate, cetyl myristate,
ceryl palmitate, ceryl certate, myricyl melissate, stearyl
palmitate, stearyl myristate, and lauryl laurate.
[0107] In a further embodiment, the invention provides a method for
controlling rate of release of a pancreatic/digestive enzyme from
an encapsulated composite upon exposure to a solvent. The method
includes coating the enzyme with an amount of an emulsifiable lipid
to form an encapsulated pancreatic enzyme substance composite,
wherein the rate of release of the enzyme from the encapsulated
composite is decreased as the amount of emulsifiable lipid based on
total weight of the encapsulated composite is increased. In the
alternative, the rate of release of the pancreatic enzyme from the
encapsulated composite is increased as the amount of emulsifiable
lipid based on total weight of the encapsulated composite is
decreased. The emulsifiable lipid useful in this embodiment can
consists essentially of one or more monoglycerides.
[0108] The solvent in which a lipid emulsifies can be an aqueous
solvent. The aqueous solvent interacts with the hydrophilic groups
present in the emulsifiable lipid and disrupts the continuity of
the coating, resulting in an emulsion between the aqueous solvent
and the lipids in the coating, thus releasing the bioactive
substance from the composites.
[0109] The methods herein, used to encapsulate pancreatic or
digestive enzyme cores for treatment of neurological conditions or
disorders, has not been previously described. The methods for lipid
encapsulation of medications for human consumption which have the
characteristics of a time-released medication, and which utilize
the lipid encapsulation for stability have not been previously
described. Prior to the experiments described herein, there was no
published protocol that allowed for the preparation of an
encapsulated enzyme preparation comprising a coating of
emulsifiable lipid and a digestive enzyme suitable for the
time-specific arid/or site-specific targeted release along the GI
tract for the treatment of autism, ADD, ADHD, Parkinson's Disease
and other neurological or behavioral conditions susceptible to
treatment with digestive enzymes.
[0110] Aspects and embodiments of the instant disclosure stem from
the surprising and unexpected discovery that certain pharmaceutical
dosage preparations comprising a coating of emulsifiable lipid and
a digestive enzyme can have novel potentiated activity and
unexpected favorable release and dissolution profiles and
absorption kinetic parameters along the various portion of the GI
tract. These characteristics are useful for formulating a specific
bioactive enzyme for site specific targeted release along the GI
tract for the treatment of autism, ADD, ADHD, Parkinson's Disease
and other neurological conditions.
[0111] Determination of whether a subject is in need of treatment
with an effective amount of digestive enzymes may be based on a
determination that the subject has an enzyme deficiency.
[0112] In one aspect of the invention, the method comprises using
the enzyme formulations of this invention to treat children and
other individuals with autism, ADD, ADHD, Parkinson's disease and
other neurological diseases or conditions, who also have an enzyme
deficiency. The enzyme deficiency could be determined by any method
used in determining or diagnosing an enzyme deficiency. In one
aspect the determination or diagnosis may be made by evaluating
symptoms, including eating habits, self-imposed dietary
restrictions, symptoms of eating disorders and/or gastrointestinal
disorders. In other aspects, the determination may be made on the
basis of a biochemical test to detect, for example, levels or
activities of enzymes secreted, excreted or present in the GI
tract, and/or by determining the presence of a mutation in a gene
or aberrant expression of a gene encoding one or more digestive
enzymes. The enzyme deficiency may also be determined, for example,
by detecting a mutation or aberrant expression of a gene encoding a
product regulating or otherwise affecting expression or activity of
one or more digestive enzymes.
[0113] In some aspects, the individual to be treated may also be
tested for the presence of a co-morbidity, which is a co-morbidity
which does not affect the activity or expression of a digestive
enzyme. In certain aspects, individuals who are determined to have
autism based on clinical symptoms but not a co-morbidity such as a
genetic co-morbidity, are treated with the enzyme delivery systems
described herein. However, individuals who are determined to have
autism based on clinical symptoms and a co-morbidity, who
nevertheless also test abnormally low for FCT level or positive
using another indicator of GI pathogens and/or low digestive enzyme
activity or expression may also be treated with the enzyme delivery
systems of this invention.
[0114] The following co-morbidities are set forth as exemplary
co-morbidities:
Fragile X
[0115] Hallermann-Streiff syndrome Trisomy 21 tranlocation on 9
Beckwith-Wiedemann syndrome
Trisomy 21
Trisomy 18
[0116] Rubenstein-Tabi syndrome
Fragile X
[0117] Prader-Willi syndrome
Trisomy 21
[0118] Rett syndrome Klippel-Feil syndrome Rett syndrome
Duchenne Muscular Dystrophy
[0119] Tourette syndrome In-utero stroke Trisomy 21
Fragile X
Juvenile RA
[0120] In-utero stroke
Trisomy6
Duchenne Muscular Dystrophy
Juvenile Diabetes
Diabetes Type I
Adrenoleukodystrophy
[0121] Wilson's disease In-utero stroke
Diabetes Type I
[0122] Prader-Willi syndrome 22q13 Tourette syndrome
Lissencephaly
[0123] Neutrophil Immunodeficiency syndrome
Diabetes Type I
[0124] Tourette syndrome
Tetrasomy 18p
[0125] Hyper IgE syndrome
Angelman Syndrome
Diabetes Type I
[0126] Rett syndrome
Fragile X
[0127] Marfan syndrome Waardenburg syndrome glutathione synthetase
deficiency
Diabetes Type I
Rubinstein-Taybi
Angelman Syndrome
Klinefelter Syndrome
[0128] Brain bleed at birth
Turner Syndrome
Hypothyroidism
Diabetes Type I
[0129] Brain damage of prematurity
[0130] In One aspect, the determination of an enzyme deficiency may
be made using a test for fecal chymotrypsin levels. Methods such as
PCR or other amplification, SNP detection, sequencing, and/or DNA
combing may be used to detect the presence of a mutation or
presence of short RNA sequences which interfere with expression of
one or more genes encoding a digestive enzyme. For example, the
mutation may in a gene encoding a digestive enzyme which decreases
or eliminates the activity of the enzyme. As another example, the
mutation may be mutation in the MET gene, a gene encoding the
pleiotropic MET receptor tyrosine kinase See Campbell et al., PNAS
103(46), 16834-39 (2006). These mutations may include, for example,
the MET promoter variant rs1858830 C allele, and or mutations in
the MET signaling pathway such as a haplotype of the SERPINE1 gene,
or the rs 344781 PLAUR promoter variant T allele.
[0131] The enzyme formulations of this invention are suited for use
in delivering digestive enzymes to individuals with autism, ADD,
ADHD, Parkinson's disease and other neurological diseases or
conditions in need of enzyme treatment. Fallon has described
certain methods and enzyme compositions for use in treating
children and other individuals, with autism, ADD, ADHD, Parkinson's
disease and other neurological diseases or conditions, for example,
U.S. Pat. Nos. 7,138,123, 6,660,831, 6,632,429, 6,534,063, hereby
incorporated by reference as if set forth in full herein.
[0132] The present invention will now be described more fully with
reference to the accompanying figures and examples, which are
intended to be read in conjunction with both this summary, the
detailed description, and any preferred and/or particular
embodiments specifically discussed or otherwise disclosed. This
invention may, however, be embodied in many different forms and
should not be construed as limited to the embodiments set forth
herein; rather, these embodiments are provided by way of
illustration only and so that this disclosure will be thorough,
complete, and will fully convey the full scope of the invention to
those skilled in the art.
[0133] In the experiments described herein, several factors were
discovered that allowed for the unexpected enhanced/potentiated
efficacy and property. For example, it was discovered that certain
encapsulation enzymatic preparations comprising soy oil exhibited
certain surprising characteristics that led to improvements in the
site-specific activity, release/dissolution profile, and ease of
manufacturing, packaging and storage. Without being bound to a
particular theory of operation, the skilled artisans will
appreciate that other methods of sample preparation and/or
formulation that can also yield these advantageous parameters are
also contemplated herein.
[0134] The following experiments describe exemplary procedures in
accordance with the invention. It is to be understood that these
experiments and corresponding results are set forth by way of
illustration only, and nothing therein shall be construed as a
limitation on the overall scope of the invention. By way of
example, these studies demonstrate some of the unexpected
improvements realized by the exemplary encapsulated enzyme
preparations of the present disclosure.
Example 1
Increased Flow Properties and Pourability of an Exemplary
Encapsulated Digestive Enzyme Preparation
[0135] Before the exemplary methods and preparations of the present
disclosure is applied, examination of an unprocessed, raw enzyme
preparation (Scientific Protein Laboratories (SPL) of Wanakee,
Wis.) revealed that it contained significant variability in
particle size and 20 irregular morphology, as shown in an electron
micrograph of the particles as pictured in FIG. 1. Some crystalline
salt particles are also visible. The raw enzyme does not pour as it
clumps and is difficult to measure due to the uneven surfaces, and
jagged edges. The raw preparation is also not suitable for lipid
encapsulation without further processing because the raw product
contains particles both too large and too small for proper
encapsulation. The sieved enzyme, while more uniform in size,
continues to exhibit uneven surfaces and clumps while pouring.
[0136] FIG. 2 shows the coated enzyme preparation produced
following sieving and lipid coating of the raw material. In this
example, the morphology of particles is significantly improved,
with rounder surfaces. This leads to a non-dusty product with good
flow and organoleptic properties.
[0137] The morphology of the enzyme is now greatly improved due to
the rounding of the surfaces, which leads to a product which is
less dusty, does not aerosolize and has good flow and improved
organoleptic properties.
[0138] The size distribution of particles in the raw enzyme
preparation is shown in the graph in FIG. 3. In general, large
particles (>40 mesh) and very small particles (<140 mesh) are
not suitable for proper encapsulation. In order to increase the
flow properties of the encapsulated pancreatic enzyme preparation,
the raw enzyme particles were sieved to include only particles of
sizes 40-140 mesh, or about 106 to 425 microns.
Example 2
Stability of an Exemplary Encapsulated Digestive Enzyme
Preparation: Temperature Storage
[0139] In a further exemplary embodiment, multiple types and weight
percentages of lipids were used to coat the sieved enzyme cores.
Properties including mechanical strength, melting point, and
hydrophobicity were taken into consideration in choosing a suitable
lipid coating for the pancreatic enzyme. Multiple examples of lipid
coatings were examined below and their physical appearances were
examined under 25.degree. C. and at 40.degree. C. Accordingly,
lipids with a range of physical properties such as mechanical
strength, melting point and hydrophobicity were evaluated for
coating of the pancreatic enzymes. In this example, it was found
that the decreasing the melting point or increasing the
hydrophilicity of the coatings were not suitable for encapsulation
because they resulted in product that would cake under accelerated
storage stability conditions. The sieved and encapsulated enzyme
preparations made using hydrogenated soy oil, hydrogenated castor
wax, and carnauba wax all demonstrated good pouring and no
caking.
[0140] Table 1 provides the results of the visible physical changes
which occurred at 25.degree. C. and 40.degree. C.:
TABLE-US-00001 Physical Appearance at Physical Appearance at
Coating System 25.degree. C. storage 40.degree. C. storage
Hydrogenated Soy oil OK OK (Balchem/Alibec) Hydrogenated Castor wax
OK OK Carnauba Wax OK OK Hydrogenated OK Some Caking Monoglycerides
Soy/Monoglcyeride blends OK Some caking
[0141] Both the hydrogenated monoglycerides and the soy
oil/monoglyceride blends demonstrated caking at the higher
temperature. Therefore it is clear that the lower melting or more
hydrophilic coatings were not suitable for encapsulation because
they resulted in a product that would cake under extended storage
conditions as evidenced by our accelerated storage condition test
at 40 degrees Centigrade.
[0142] Both the hydrogenated monoglycerides and the soy
oil/monoglyceride blends demonstrated caking at the higher
temperature. Therefore it is clear that decreasing the melting
point or increasing the hydrophilicity of the coatings were not
suitable for encapsulation because they resulted in a product that
would cake under extended storage conditions as evidenced by our
accelerated storage condition test at 40 degrees Centigrade.
Example 3
An Exemplary Encapsulated Digestive Enzyme Preparation Suitable for
Pancreatic Enzymes: Enzyme Activity Measured as a Function of
Stability
[0143] In a further embodiment, enzyme stability was determine
according to the following method: For the accelerated test,
standard ICH guidelines were used: the coated preparations were
placed in a plastic container, which was stored in a controlled
humidity cabinet at 40.degree. C. and 75% relative humidity.
Enzymatic activity was measured by grinding the coated enzyme
preparations, dispersing in appropriate buffers, and testing for
lipase activity.
TABLE-US-00002 TABLE 2 PERCENT STABILITY OF ENCAPSULATED ENZYMES
WHEN STORED AT 40 C./75% RH, IN CLOSED CONTAINERS Activity Activity
Activity Activity 1 week 2 weeks 1 Month Sample Lot# or coat RT
Capped Capped capped PEC raw Nov '06 1206-1369A 116% 126% 75% PEC
encap 70%, R1C-0890 118% 112% monoglyceride PEC encap 50%, R1C-0891
116% 110% 88% soy/mono PEC raw Jan '07 1206-1382B 113% 61% PEC
encap 70% R1C-0898 carnuba PEC encap 50% R1C-0898 68% carnuba PEC
encap 70%, castorwax 108% 78% 87% castor wax PEC encap 80%, soy 99%
89% 87% soy
[0144] As illustrated above in the data summarized in Table 2, the
soy oil 80% appeared to impart the greatest amount of stability of
all the lipids, an effect that surprisingly was greater for enzyme
preparations stored in capped containers than in uncapped
containers. Tests of stability for 75% relative humidity enzyme
preparations stored at 40.degree. C. in open pans did not show
significant differences in stability between coated and uncoated
preparations.
Example 4
An Exemplary Encapsulated Digestive Enzyme Preparation Suitable for
Pancreatic Enzymes: Enzyme Activity And Rate of Release of Multiple
Soy Encapped Pancreatic Enzyme
[0145] In a further embodiment, encapsulates were prepared
according to the methods described below. The raw enzyme material
was sieved to obtain particles smaller than 40 mesh but larger than
140 mesh, to remove fines, and to obtain a more uniform mixture
more suitable for enteric coating.
[0146] The following preparations were made:
70% active enzyme by weight, with a standard stable soy coating;
80% active enzyme by weight, with a standard stable soy coating;
and 90% active enzyme by weight, with a standard stable soy
coating.
[0147] Activity in each encapsulated enzyme preparation was
measured by grinding the encapsulates, dispersing the ground
material in appropriate buffers, and testing for lipase
activity.
[0148] As shown in FIG. 4, the enzyme activity in the coated
preparations does not show any significant loss of activity upon
coating (decrease from 110 to 100% activity, normalized to stated
enzyme activity of the raw enzyme material).
[0149] Enzyme release was measured by suspending each encapsulate
in a dissolution apparatus at pH 6.0 buffer for 30, 60, and 90
minutes (100 rpm, as per USP guidelines). As shown in FIG. 5, all
encapsulates show between 80-90% release at 30 and 60 minutes. At
90 minutes, the measured enzyme activity obtained with these
preparations decreases.
Example 5
An Exemplary Encapsulated Digestive Enzyme Preparation Suitable for
Pancreatic Enzymes: Particle Size of Multiple Soy Oil Encapped
Pancreatic Enzyme
[0150] In a further embodiment, preparations containing 70% or 80%
active pancreatic enzyme by weight, encapsulated with soy oil were
compared to raw pancreatic enzyme material with respect to particle
size, as shown in FIG. 6.
[0151] All levels of lipid demonstrate an impact of particle size.
The 80% PEC demonstrates the most uniform as none appear at the 200
mesh level.
Example 6
An Exemplary Encapsulated Digestive Enzyme Preparation Suitable for
Pancreatic Enzymes: Smell and Taste
[0152] Examination of exemplary encapsulated enzyme preparations
containing 70%, 80% and 90% enzyme by weight were performed to
determine their taste and smell when compared to Sucanat.TM. and
brown sugar, as well as compared to the raw enzyme. The results are
shown in Table 4, below. Sucanat.TM. is an organic whole food
sweetener.
TABLE-US-00003 TABLE 4 SUBSTANCE ODOR TASTE Brown sugar Yes Sweet
Sucanat .TM. No Sweet Raw Enzyme Meaty/smoky N/A 70% No No 80% No
No 90% Slight Salty
Example 7
An Exemplary Encapsulated Digestive Enzyme Preparation Suitable for
Pancreatic Enzymes: Manufacturing
[0153] The flow chart outlining the manufacturing process useful in
making the enzyme preparations of this invention is shown in FIG.
7.
[0154] Ingredients used in making a batch of an exemplary
encapsulated pancreatic enzyme preparation included 20.0 lb. of
sieved pancreatic enzyme and 5.0 lb. of hydrogenated vegetable oil,
for example, soy oil.
[0155] The pancreatic enzyme concentrate was first sieved through a
40 USSS mesh screen, and the material which passed through the mesh
was retained. The retained material was then screened through a 140
USSS mesh screen (or the equivalent), and the material which did
not pass through the mesh was retained as the sieved pancreatic
enzyme material or particles.
[0156] In the encapsulation process, the appropriate coating
material is charged to the melt pot, and brought to and maintained
at 110.degree. F. for the spraying process. Any temperature that
will provide appropriate consistency during the spraying process
may be used. In some embodiments, the temperature is further
selected based on the melting points of the lipids used in the
coating, and/or so that after contact of the sieved pancreatic
enzyme material or particles with the coating, the activity of the
enzyme preparation remains about the same.
[0157] The liquefied coating material is weighed and transferred to
the spray pot. The sieved pancreatic enzyme was added to the
encapsulation manufacture vessel. The pancreatic enzyme particles
are encapsulated with coating material to the selected coating
level.
[0158] The encapsulated material is screened with a 14 USSS mesh
screen (or equivalent), and the material that passes through the
screen is retained. Following sieving, the material is collected
and samples are removed for QC.
[0159] If two sub-batches are to be blended, the loaded screened
material is added to a suitable blender and blended for 7 to 10
minutes. Samples are obtained for finished product testing. The
encapsulated material is bulk packaged and placed in quarantine
pending test results. Upon achieving acceptance criteria, the
finished product is released by the Quality group. Afterwards, the
product may be shipped as directed.
[0160] Samples are collected for finished product testing,
including analytical testing and microbial assays, which can be
tested over time.
Example 8
An Exemplary Encapsulated Digestive Enzyme Preparation Suitable for
Pancreatic Enzymes: Packaging
[0161] In yet another further embodiment, the stability of the
enzyme is due in part to the encapsulation and in part to the
trilaminar foil packaging. The following demonstrates the packaging
process for the single dose sachets/pouches.
[0162] First, following manufacture the product is dispensed into
clean, drums double lined with food-grade polyethylene bags, and
the drums are sealed. If specification criteria are met, the lot is
then released from quarantine, and the material is then shipped to
a suitable packager for placement into sachets for individual
dosing to the patient.
[0163] For example a PD-73272 Printed Child Resistant (CR) Pouch
consisting of 26#C1S Paper/7.5# LDPE/0.0007'' Aluminum Foil/15#
with a Surlyn liner is utilized for packaging. Preferably
pre-printed film/foil, exterior printing will be with 1 color
eye-mark on white background while in-line printing of lot number,
expiration date and product code will also be in 1 color, black.
Overall sachet dimension are: W 2.50''.times.H 3.50''. The sachet
is sized to hold 900 mg of granules of Pancrelipase
lipid-encapsulated drug product with a tolerance of .+-.10% into a
unit dose pouch/sachet. The final product will have a protease
activity of not less than 156 USP units/mg.
Example 9
An Exemplary Encapsulated Digestive Enzyme Preparation Suitable for
Pancreatic Enzymes: Dissolution
[0164] The effect of the release of Pancreatase from lipid
encapsulated particles with soy oil was studied using particles
with varying levels of lipid coating (expressed as % lipid coating
per total particle weight. The coating level was varied from 10% to
30%. There was no significant effect of lipid coating in this range
on the release of pancreatase in an aqueous environment from the
particles over a 60-minute period. All formulations release over
80% of the enzyme within the first 30 minutes following the
initiation of dissolution. Maximum release for the 90%, 80% and 70%
particles was 85%, 88% and 83% respectively by 60 minutes.
Example 10
An Exemplary Enzyme Delivery System for Treatment of Autism
[0165] The choice of 70%-90% encapsulated pancreatic enzyme
preparation (active enzyme by weight) was selected on the basis of
its release profile, as suitable for release of the enzyme in the
proximal small intestines where protein digestion by the protease
component will take place.
[0166] Soy oil was selected as the lipid coating, for its lack of
protein components, and corresponding lack of antigenic properties,
to minimize or eliminate the possibility of an allergic reaction to
the lipid coating in treated patients and children with autism.
[0167] The use of the 70-90% preparation increases pourability and
flow properties while decreases aerosolization, which permits use
of a sachet or pouch delivery system.
[0168] The addition of the trilamminar foil housing insures that
the sprinkle formulation will be stable, transportable, and will be
delivered by a single unit dose mechanism.
[0169] The low lipase formulation allow also for the safety by
reducing the potential for colonic strictures, and enhances the
utilization of the protease portion of the enzyme.
TABLE-US-00004 TABLE 5 Composition of LUMINENZ-AT encapsulated
digestive enzyme preparation, 900 mg Sachets Compendial Ingredients
Status Functions Content Pancreatic enzyme USP Active ingredient
NLT 156 USP concentrate (porcine units/mg origin) Hydrogenated NF
Lipid coating q.s. vegetable oil, Type I material (soybean oil)
[0170] The drug substance, pancreatic enzyme concentrate (porcine
origin) is purchased from an appropriate supplier. The properties
of the pancreatic enzyme concentrate (pancreatin/pancrelipase)
suitable for use in the products of this invention are described in
the table below.
TABLE-US-00005 TABLE 6 Parameter USP Specification Protease (USP)
NLT 25 USP Units/mg Lipase (USP) NLT 2 USP Units/mg Amylase (USP)
NLT 25 USP Units/mg Fat (USP) NMT 6.0%* Loss on Drying (USP) NMT
5.0% Escherichia coli (USP) Neg/10 g Salmonella species (USP)
Neg/10 g *If less than 75 U/mg Protease, 6 U/mg Lipase or 75 U/mg
Amylate, then specification is NMT 3.0%
Specifications for Hydrogenated Vegetable Oil (Soy Oil)
Physical Appearance and Sensory Characteristics:
[0171] Material provided in flake or powder form, free from foreign
matter and objectionable odor.
TABLE-US-00006 TABLE 7 Specification Analytical Procedure Chemical
Parameter Melting Range 67 to 69 C USP/NF <741> Class II Acid
Value 0.4 Max. USP/NF <401> Iodine Value 5.0 Max. USP/NF
<401>Method II Loss on drying 0.1% Max. USP/NF <731>
Saponification Value 175-200 USP/NF <401> Heavy Metals 0.001%
Max. USP/NF <231> Organic Volatile Complies USP/NF
<467>Method IV Impurities Residual Solvents Complies USP/NF
<467> Unsaponifiable Matter 0.8% Max. USP/NF <401>
Microbial Parameters Total Aerobic Microbial 2000 cfu/g max. USP/NF
<61> Count Staphylococcus aureus Absent in 10 g USP/NF
<61> Pseudomonas seruginosa Absent in 10 g USP/NF <61>
Salmonella Species Absent in 10 g USP/NF <61> Escherichia
coli Absent in 10 g USP/NF <61> Mold and yeast 200 cfu/g max.
USP/NF <61>.
[0172] Chromatograms of working standard, diluent, mobile phase B,
and placebo demonstrate no interference with the standard peak (see
Chromatogram, FIG. 9). The analytical placebo and active tablet
compositions are given in Table 8.
TABLE-US-00007 TABLE 8 Composition of Analytical Placebo and Active
Powder Analytical Placebo Active Formulation Ingredients mg/sachet
mg/sachet Pancreatic enzyme -- 720.0 concentrate Lipid encapsulate
180.0 180.0 Total 180.0 900.0
[0173] The method linearity was evaluated by analyzing several
sample levels of the standard concentration in the presence of the
placebo matrix. These levels were 50%, 70%, 100%, 130%, and 150%.
Three injections of each sample were used to calculate the average
response (area/concentration) for that level. Then the relative
standard deviation for the generated response ratios was calculated
along with the least-squares linear regression statistics for the
average peak area vs. concentration (see Tables 9 and 10). A plot
of the average peak area vs. concentration with the linear
regression line is given in FIG. 10.
TABLE-US-00008 TABLE 9 Linearity Data Average Standards Peak mg/
Peak Area (mAU s) Area Average mL % Injection 1 Injection 2
Injection 3 (mAU s) Response 0.100 50 709.4 712.9 710.5 710.9
7109.3 0.140 70 1041.2 1040.0 1002.9 1028.0 7343.1 0.200 100 1529.0
1499.0 1523.1 1517.0 7585.2 0.260 130 1969.4 2010.3 1996.2 1992.0
7661.4 0.300 150 2336.2 2322.6 2350.6 2336.5 7788.2
TABLE-US-00009 TABLE 10 Linearity Results Parameter Criterion
Result Correlation Coefficient >0.997 0.999 y Intercept .+-.2.0%
-112.9 RSD of Response Ratios <2.0% 3.6 Visual Linear yes
Standard error of y intercept -- 18.8 Slope -- 8139.1
Example 11
Biochemical Biomarkers, and Behavioral Core and Non-Core Symptoms
of Autism
[0174] The correlation between digestive enzyme deficiencies in
autistic children was determined in children diagnosed with autism
based on clinical (behavioral) symptoms. This correlation was also
studied in children diagnosed with autism and a genetic
co-morbidity. Following the initial discovery that autistic
children exhibited self-imposed protein dietary restrictions,
studies were conducted which indicated that abnormally low levels
of fecal chymotrypsin (FCT) is useful as a biomarker for
autism.
[0175] In addition, the number of autistic patients responding to
pancreatic enzyme replacement was also determined, based on
biomarker measurements and clinical symptoms. Changes in the
gastrointestinal system as well as a change in the core symptoms of
autism were examined. The table below provides an overview of the
studies conducted at multiple physician based sites.
TABLE-US-00010 TABLE 11 Study Number Total # of Subjects Autism
Non-autism 1 9 9 2 26 26 3 46 25 21 4 54 54 5 463 266 197 6 320 64
256 7 33 33 8 42 25 17 9 68 68 10 225 225 11 175 175
[0176] Initial observations were based on observation of
self-imposed dietary restriction by almost all children with
autism. Multiple studies were then conducted to evaluate the
ability of autistic children to digest protein. A study of the
physiology of protein digestion led to an examination of the
gastrointestinal system's cascade of digestive enzymes, especially
those involved in protein degradation, such as chymotrypsin. As a
measure of dysfunction, it was determined that fecal chymotrypsin
(FCT) levels in children suffering from autism were abnormally
low.
Study 1
[0177] This initial study was an exploratory one to determine if a
small cohort of children with autism indeed would have abnormally
low levels (<9.0) of fecal chymotrypsin. (FCT). The results of
study 001 is shown in FIG. 10
[0178] All 9 children with autism evidenced an abnormally low FCT
level of below 7 Units/gram. (Normal.gtoreq.9.0). This observation
in a small set of children led to further examination of the
potential for a physiological link to autism heretofore
undiscovered.
Study 2
[0179] Study 2 was undertaken to determine if a larger cohort of
children (26 children) with autism also experienced abnormally low
FCT levels. Levels of fecal elastase-1, another pancreatic
digestive enzyme present at low amounts in pancreatic
insufficiency, was also determined. Again, the levels of FCT were
abnormally low in 25 of the 26 children, falling at 8 U/g or below.
One child had an FCT level of 9 U/g. On the other hand, all of the
children had normal levels of fecal elastase-1.
Study 3
[0180] In Study 3, FCT levels were determined in 46 children aged 2
years to 14 years of age, 25 with autism and 21 without autism, The
data demonstrated that the children with autism had abnormally low
FCT levels and those children who did not have autism had normal
FCT levels, of 12 U/g or higher. The results are summarized in FIG.
12. The top line in FIG. 12 shows the FCT levels in subjects who
did not have autism, while the bottom line shows the FCT levels in
subjects who did have autism.
Study 4
[0181] In Study 4, 54 children diagnosed with autism and a
co-morbid genetic disorder were examined for FCT levels. The data
showed that the children with autism and a co-morbid genetic
disorder tested normal for FCT level.
[0182] As autism is determined by behavioral assessment, it was
hypothesized that autism due to, or present with a known genetic
disorder may have a differing physiology from others with autism
alone, or not due to a known genetic disorder. Some genetic
disorders have typical symptoms, while others may be more variable
and overlap with autistic symptomology. This study examined
children with autism who were also diagnosed with another known
condition, to determine if FCT levels were abnormally low in these
children.
[0183] Table 12 below represents 54 children diagnosed with autism
who also had a genetic co-morbidity
TABLE-US-00011 TABLE 12 Children Diagnosed with Autism who also
Have a Genetic Co-Morbidity FCT Level Co- U/g Morbidity 1 12
Fragile X 2 22 Hallermann-Streiff syndrome 3 25.2 Trisomy 21 4 15.8
tranlocation on 9 5 18 Beckwith-Wiedemann syndrome 6 26.6 Trisomy
21 7 39.2 Trisomy 18 8 16.6 Rubenstein-Tabi syndrome 9 25.4 Fragile
X 10 20.6 Prader-Willi syndrome 11 14.6 Trisomy 21 12 25.6 Rett
syndrome 13 21.4 Klippel-Feil syndrome 14 20.6 Rett syndrome 15
24.8 Duchenne Muscular Dystrophy 16 12.2 Tourette syndrome 17 14.8
In-utero stroke 18 30 Trisomy 21 19 18.8 Fragile X 20 17.6 Juvenile
RA 21 18.8 In-utero stroke 22 34 Trisomy6 23 22.2 Duchenne Muscular
Dystrophy 24 18.8 Juvenile Diabetes 25 28.4 Diabetes Type I 26 13.8
Adrenoleukodystrophy 27 44 Wilson's disease 28 19.6 In-utero stroke
29 7.4 Diabetes Type I 30 23.4 Prader-Willi syndrome 31 14.4 22q13
32 15.4 Tourette syndrome 33 17.6 Lissencephaly 34 22.4 Neutrophil
Immunodeficiency syndrome 35 18.4 Diabetes Type I 36 32.2 Tourette
syndrome 37 14.6 Tetrasomy 18p 38 31 Hyper IgE syndrome 39 26.6
Angelman Syndrome 40 17.4 Diabetes Type I 41 12.6 Rett syndrome 42
34 Fragile X 43 17.4 Marfan syndrome 44 21.2 Waardenburg syndrome
45 21.8 glutathione synthetase deficiency 46 6.0 Diabetes Type I 47
26.6 Rubinstein-Taybi 48 34 Angelman Syndrome 49 25.2 Klinefelter
Syndrome 50 21.4 Brain bleed at birth 51 16.8 Turner Syndrome 52
23.4 Hypothyroidism 53 15.8 Diabetes Type I 54 7.8 Brain damage of
prematurity
[0184] Only two of the 54 children diagnosed with both autism and a
genetic co-morbidity had abnormally low levels of FCT. Those
children had Type I diabetes. 52 of the 54 children registered FCT
levels in the normal range.
[0185] This further supports that low FCT levels are present in
children diagnosed with 5 autism in the absence of another known
genetic morbidity.
Study 5
[0186] In Study 5, FCT levels were determined for 463 children aged
2 years to 8 years of age, 266 diagnosed with autism and 197
diagnosed without autism, in a multi-office physician-conducted
study. The data showed that the children with autism had abnormally
low fecal chymotrypsin levels and those children who did not have
autism had normal levels of fecal chymotrypsin.
[0187] The data is summarized in table 13 below.
TABLE-US-00012 TABLE 13 Mean Fecal Chymotrypsin Levels in Children
with and without Autism Children with Children not N = 463 Autism
with Autism Total numbers of children 266 197 Mean FCT (U/g) 4.4
23.2 Total Children with Abnormal 203 3 Levels of FCT % (p <
0.001) 76.34% 1.50% Total Children with Normal 63 194 Levels of FCT
(p < 0.01) % 23.68% 98.50%
[0188] This data further established that children diagnosed with
autism who do not also have a known genetic co-morbidity have
abnormally low levels of FCT. FCT levels may therefore be useful in
diagnosing children with autism, if the child does not also have a
known genetic co-morbidity (unless the co-morbidity is Type I
diabetes).
[0189] Chymotrypsin is a pancreatic enzyme. Chymotrypsin is a
serine protease and is unique in that it cleaves only essential
amino acids during the digestive process. Specifically,
chymotrypsin cleaves the peptide bond on the carboxyl side of
aromatic amino acids. A lack of protein digestion as evidenced by
abnormal FCT levels leaves the child with a dearth of amino acids
available for new protein synthesis. Without sufficient levels of
essential amino acids, new proteins required for various bodily
functions cannot be synthesized. For example, a shortage or lack of
proteins involved in neurological processes may then give rise to
symptoms of autism.
Study 6
[0190] In Study 6, FCT levels were determined for 320 children aged
2 years to 18 years of age, 64 with autism, 64 with ADD. 64 with
ADHD, 64 with known genetic conditions, and 64 normals (no known
conditions). The data showed that the children with autism, ADD and
ADHD exhibited abnormally low levels of FCT compared to the
children with known genetic conditions and normal children. FCT
data were gathered during a multi-physician office trial of
age-matched children with multiple conditions. FIG. 13 depicts FCT
levels in separate groups of children aged 6 years to 18 years who
have Autism, ADHD (Attention Deficit Hyperactivity Disorder), ADD
(Attention Deficit Disorder), known genetic disorder also diagnosed
with autism, or no known condition (normals).
[0191] The two upper lines in FIG. 13 correspond to FCT levels in
children without any known condition and children with known
co-morbid conditions (genetic and others). The three bottom lines
correspond to FCT levels in the children with autism, ADD and
ADHD.
[0192] The Autism, ADD, and ADHD children had significantly lower
levels FCT than those without any known condition, or those with a
known genetic co-morbidity or traumatic condition (p<0.01).
Study 7
[0193] In Study 7, 33 children who were diagnosed with autism and
abnormally low FCT levels were enrolled in the study. The children
were treated with one of two pancreatic/digestive enzyme
supplements, or given no treatment. FCT levels were measured for
each child at time 0, 30, 60, 90 and 120 days.
[0194] Eleven (11) children were given a low therapeutic dose of
ULTRASE.RTM. MT20 (pancrelipase) Capsules (opened to sprinkle on
food) (see below); 11 children were given Viokase.RTM.
(pancrelipase) powder for sprinkling on food at a minimal dosing
level of/teaspoon; 11 children just had their fecal chymotrypsin
levels measured. All children were age-matched and without a
co-morbid neurological and/or genetic diagnosis.
[0195] Each ULTRASE Capsule was orally administered and contained
371 mg of enteric-coated minitablets of porcine pancreatic
concentrate contained:
TABLE-US-00013 Lipase 20,000 U.S.P. Units Amylase 65,000 U.S.P.
Units Protease 65,000 U.S.P. Units
Each 0.7 g (1/4 Teaspoonful) of Viokase Powder contained: Lipase,
USP units 16,800 Protease, USP units 70,000 Amylase, USP units
70,000
[0196] FCT levels were monitored over 120 days to determine whether
FCT levels changed in response to treatment with either of the
enzyme formulations, compared to the children who did not receive
enzyme treatment. The results of the FCT levels, measured over a
120 day period are shown in Table 14 below.
TABLE-US-00014 TABLE 14 Mean Fecal Chymotrypsin Levels at Baseline,
30, 60, 90 and 120 days Post administration of Multiple Pancreatic
Enzyme Replacement Ultrase Viokase No treatment Mean FCT (units)
3.49 3.81 3.1 At Baseline Mean FCT (units) 5.05 7.02 3.15 30 Days
Mean FCT (units) 4.82 8.96 3.18 60 Days Mean FCT (units) 4.91 13.73
3.25 90 Days Mean FCT (units) 5.38 15.1 3.13 120 Days N = 33
[0197] The results are shown in the bar graph in FIG. 14. The top
bar (very pale bar) for each time point shows the FCT level for the
untreated children. The middle bar shows the FCT level for children
treated with Viokase, and the bottom bar at each time point shows
the FCT level following Ultrase treatment. The results in the table
and graphed in FIG. 14 indicate that a significant change in FCT
level was seen only following administration of the Viokase
enteric-coated enzyme formula, from baseline at time 0 to 120 days.
The greatest change was seen in the first 90 days. The changes in
the first 90 days were significant compared to the changes seen
between 90 and 120 days. While the Ultrase group showed some change
from baseline to 120, the change was not significant.
[0198] The lipases in Ultrase are very sensitive to pH changes and
to degradation in acidic conditions, such as those found in the
stomach. The enteric coating on Ultrase allows the enzymes to
bypass the stomach. Ultrase has been shown to be useful for
delivery of sufficient lipases to treat adults with cystic fibrosis
and chronic pancreatitis who suffer from pancreatic enzyme
deficiency. However, the enteric coating on Ultrase and other
similar products apparently did not allow the protease portion of
those compositions to be delivered in the proximal small intestine,
where it is needed for protein degradation. As demonstrated in the
small pilot study, Ultrase did not allow for release of the
protease portion of the enzyme, specifically chymotrypsin, as
determined by FCT levels measured following administration of
Ultrase. The FCT levels in the Ultrase treated group were similar
to those found in the NO TREATMENT group.
[0199] The optimum delivery timing and location for the protease
portion of the enzyme is from the latter portion of the time the
bolus of food is in the stomach, through the time the digesting
food spends in the proximal small intestine.
Study 8
[0200] In Study 8, 42 age-matched children, 25 with autism, and 17
without autism or other co-morbid condition, were examined using a
stool test for the presence of multiple pathogens as well as
markers of Gastrointestinal dysfunction, including FCT levels. The
children with autism had a larger number of stool pathogens present
as well as abnormally low FCT levels.
[0201] This small pilot study was undertaken to examine the
gastrointestinal flora of children with autism versus those without
autism. Multiple markers of gastrointestinal health were examined
to determine if there is an abnormal gastrointestinal presentation
in these children.
[0202] 42 children aged matched 25 with autism and 17 without
autism or other co-morbid condition were screened using a stool
test for the presence of multiple pathogens as well as markers of
Gastrointestinal dysfunction. Other GI pathogens or stool markers
known to those of skill in the art may also be tested as a marker
of GI dysfunction. Table 15 below shows the incidence of presence
of a GI pathogen or other stool marker.
TABLE-US-00015 TABLE 15 Incidence of the Presence of Pathogens and
other Stool Markers Representing Gastrointestinal Dysfunction % NOT
% AUTISM TOTAL AUTISM TOTAL LOW FCT 25 100% 0 0% C. difficile
antigen 15 60% 1 6% Fecal Elastase <200 0 0% 0 0% H. pylori
antigen 17 67% 0 0% E. Histolytica antigen 8 32% 0 0% Giardia
antigen 9 36% 1 6% Yeast overgrowth 4 16% 0 0% Cryptosporidium 9
36% 1 6% N = 25 N = 17
[0203] The presence of positive stool markers in the children with
autism, including low levels of fecal chymotrypsin indicated
additional gastrointestinal problems in patients with autism.
Study 9
[0204] In Study 9, 68 children aged 3 years to 8 years of age,
diagnosed with autism who presented with abnormally low FCT levels
were administered a combination of pancreatic/digestive enzymes for
90 days. Results demonstrated significant improvement in 5 out of 5
areas representing both the core and non-core symptoms of
autism.
[0205] Examination of the multiple areas of symptomology in the
children with autism in this study included both gastrointestinal
symptoms as well as the core symptoms of autism. It is well
documented in the literature that children with autism do not
change over time, and that their level of autism is static
regardless of the age of the child. Further there is thought to be
no maturation changes accompanying those with autism.
[0206] In this study, 68 children aged 3-8 diagnosed with autism
who presented with abnormally low FCT levels were administered 1/4
teaspoonful of Viokase, and a chewable papaya enzyme (Original
Papaya Enzyme Brand) at each meal for a period of 90 days.
Original Papaya Enzyme
Supplement Facts
Serving Size: 3 Tablets
Servings Per Container: 33
TABLE-US-00016 [0207] Carbohydrates <1 g <1% Sugars <1 g
Papain 45 mg ** Amylase 6 mg ** Protease 6 mg ** Papaya Fruit
(Carica 3 mg ** papaya) * Based on a 2,000 calorie diet
**Daily Values not established
[0208] The physician and the parent were asked to complete a rating
scale for each of the symptoms examined in the study. Each symptom
was rated on the scale below with (0) indicating that the child is
able to perform the task, thereby demonstrating no impairment, to
(10) representing the child's complete inability to perform the
task. With respect to undesirable behaviors such as hyperactivity
or obsessive compulsive behavior, a change from a lower score to a
higher score indicates an improvement, because the child is
demonstrating the undesirable behavior less often. The rating scale
was as follows:
[0209] Child experiences a 0% ability to perform this task
[0210] 9 Child can perform this task 10% of the time
[0211] 8 Child can perform this task 20% of the time
[0212] 7 Child can perform this task 30% of the time
[0213] 6 Child can perform this task 40% of the time
[0214] 5 Child can perform this task 50% of the time
[0215] 4 Child can perform this task 60% of the time
[0216] 3 Child can perform this task 70% of the time
[0217] 2 Child can perform this task 80% of the time
[0218] 1 Child can perform this task 90% of the time
[0219] 0 Child can perform this task 100% of the time
[0220] The average of the two scores taken at each interval:
baseline and 90 days. The scores obtained are shown in Table 16
below:
TABLE-US-00017 TABLE 16 Symptom Scores for Children with Autism
Pre- and Post-Administration of Digestive Enzymes Sum of Total
Patient Mean Sum Total Scores Score Patient Scores Mean Score Pre-
Pre- 90 Days Post 90 Days Post Digestive Digestive Enzyme Enzyme
Enzyme Enzyme Admin Admin Hyperactivity 300 4.41 568 8.35 Obsessive
255 3.75 554 8.15 Compulsive Behavior Eye Contact 552 8.12 206 3.03
Speech 553 8.13 223 3.28 Partial 515 7.57 197 2.9 Toilet Training N
= 68
[0221] CARS scores have been used to study core symptoms of autism.
In study 9, measures of core and non-core symptoms of autism were
obtained (hyperactivity, obsessive compulsive behavior, eye
contact, speech, partial toilet training) While the diagnosis of
autism was made strictly on the basis of a behavioral assessment of
the core symptoms of autism, the study indicates that other
non-core symptoms such as a lack of toilet training, will lead to
significant morbidity in this population. The 5 parameters measured
in this study indicated that the increase in toilet training, eye
contact, and speech as well as the decrease in hyperactivity and
obsessive compulsive behaviors are core and non-core symptoms that
were improved by treatment with digestive enzymes.
Study 10 and Study 11
[0222] In Studies 10 and 11, 225 children ages 2-4 years of age,
and 171 children 5-11 years of age each of whom presented with
abnormally low levels of fecal chymotrypsin, were administered a
combination of pancreatic/digestive enzymes 3 times a day for a
period of 150 days. Nine total measures of autistic symptomotology,
both core and non-core, were obtained at baseline and over a period
of 150 days. Significant changes representing improvements in both
core and non-core symptoms were seen across all age levels, with
the greatest change taking place over the first 90 days.
[0223] Each of these studies were conducted similar to the protocol
in STUDY 9. The children were divided into age groups of 2-4 and
5-11. In these studies, 225 children aged 2-4 and 171 aged 5-11
previously diagnosed with autism who presented with abnormally low
fecal chymotrypsin levels were administered 1/4 teaspoonful of
Viokase, and a chewable papaya enzyme (Original Papaya Enzyme
Brand) at each meal for a period of 150 days. The same rating scale
used in STUDY 9 was utilized in these two studies. Additionally
levels of toilet training, hand flapping, play habits, and formed
bowel movements were assessed. The % of the cohorts that
experienced changes were calculated as well. This study was
extended to 150 days, with no significance seen between day 90 and
day 150.
[0224] Table 17 below shows the measurements obtained for the
percentage of children in each group who exhibited the indicated
trait or behavior, including hyperactivity, obsessive compulsive
behavior, hand flapping, eye contact, speech, partial toilet
training, full toilet training, formed bowel movement and playing
well with others.
TABLE-US-00018 TABLE 17 PERCENT (%) WITH TRAIT OR SYMPTOM FOLLOWING
ENZYME REPLACEMENT Aged 2-4, N = 225 Aged 5-11, N = 171 Therapy Day
Therapy Day Day Day Day Day Day Measure 0 Day 60 150 0 60 150 Had
some eye contact 4 61 88 14 59 89 Had some speech 23 58 75 18 64 86
Were partially toilet trained 8 61 75 11 47 72 Were fully toilet
trained 4 30 45 16 16 20 Had formed bowel 15 88 100 16 18 97
movement Experienced hyperactivity 85 38 19 98 51 33 Plays well
with others 12 38 60 36 43 71 Experienced hand flapping 81 46 31 1
75 36 Experienced other OCD 90 73 32 91 58 22
[0225] In studies 9, 10, and 11, measurements of core and non-core
symptoms of autism were obtained. While the diagnosis of autism has
been made strictly as a result of a behavioral assessment of the
core symptoms of autism, other non-core symptoms lead to
significant morbidity in this population. The lack of toilet
training and formed bowel movements, for example, create a hardship
for parents, and often lead to a lack of social integration,
further contributing to the core symptoms of autism. This
additional isolation due to the non-core symptoms of autism further
impedes the child's ability to learn and to integrate socially.
This dynamic is continually present in this population. This effect
can be a significant driver of the core symptoms of autism. This
demonstrates that these non-core symptoms may also be valuable as
indicators of autism.
Example 12
Enzyme Delivery System Used in the Treatment of Autism
[0226] Encapsulated digestive enzyme preparations according to this
invention are packaged in pouches containing 900 mg/pouch, and are
administered to a patient in need thereof by sprinkling the
contents of one pouch onto food just before serving, administered
three times per day. Determination of whether a patient is in need
of administration of treatment with digestive enzymes including
encapsulated digestive enzyme preparations such as those of this
invention can be made using any test or indicator that is useful as
a marker of a digestive enzyme deficiency. This determination is
made, for example, using FCT levels, behavioral symptoms (core or
non-core symptoms of autism), or detection of a mutation in a gene
affecting the activity and/or expression of digestive enzymes, for
example, a MET gene mutation.
[0227] Relevant symptoms of the patient's condition or disease are
measured before and following a period of treatment. The percentage
of patients exhibiting some eye contact, some speech, partial
toilet training, full toilet training, formed bowel movements, and
ability to play well with others increases at 60 days, or earlier
than 60 days, with a further increase at 150 days. The changes
observed upon treatment with the digestive enzymes of this
invention take place over a shorter time course, and/or result in
greater improvement in each individual at any given time point
and/or improvements in core and non-core symptoms in a higher
percentage of individuals treated. In addition, a corresponding
increase in the number of patients exhibiting a decrease in
hyperactivity, hand flapping, or another OCD is observed at 60
days, with a further increase in the number of patients exhibiting
a decrease in those behaviors at 150 days.
[0228] Other core symptoms of autism such as those measured in a
CARS test are also observed and shown to improve following
treatment.
* * * * *