U.S. patent application number 16/883297 was filed with the patent office on 2020-09-10 for methods and compositions for the prevention and treatment of influenza.
The applicant listed for this patent is Curemark, LLC. Invention is credited to James Fallon, Joan M. Fallon, Matthew Heil, James Szigethy.
Application Number | 20200282030 16/883297 |
Document ID | / |
Family ID | 1000004856742 |
Filed Date | 2020-09-10 |
United States Patent
Application |
20200282030 |
Kind Code |
A1 |
Fallon; Joan M. ; et
al. |
September 10, 2020 |
Methods and Compositions for the Prevention and Treatment of
Influenza
Abstract
This disclosure relates to the prevention and treatment of
Influenza, and more particularly Influenza A Virus Subtype H1N1,
with the use of a pharmaceutical composition comprising one or more
digestive enzymes, such as pancreatic enzymes and porcine
pancreatic enzymes. The disclosure further relates to the use of an
individual's fecal chymotrypsin level as an indicator, e.g.,
biomarker of whether an individual may be more susceptible to
Influenza, e.g., Influenza A Subtype H1N1, and/or whether an
individual will benefit from administration of the described
pharmaceutical compositions. Use of the compositions as sanitizers,
antiseptics, disinfectants, and detergents, e.g., to reduce or
eradicate influenza virus present on living or inanimate surfaces
is also contemplated.
Inventors: |
Fallon; Joan M.; (White
Plains, NY) ; Heil; Matthew; (Sherman, CT) ;
Fallon; James; (Armonk, NY) ; Szigethy; James;
(Montgomery, NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Curemark, LLC |
Rye Brook |
NY |
US |
|
|
Family ID: |
1000004856742 |
Appl. No.: |
16/883297 |
Filed: |
May 26, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15265620 |
Sep 14, 2016 |
10716835 |
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16883297 |
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13502989 |
Jun 28, 2012 |
9511125 |
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PCT/US10/53484 |
Oct 21, 2010 |
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15265620 |
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61253805 |
Oct 21, 2009 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 9/0014 20130101;
A61K 39/145 20130101; A61K 38/54 20130101; C12N 7/00 20130101; C12N
2760/16134 20130101; A61K 38/4826 20130101; A61K 2039/5252
20130101; A61K 9/0053 20130101; A61K 38/465 20130101; A61K 38/4873
20130101; A61K 2039/5254 20130101; A61K 38/47 20130101 |
International
Class: |
A61K 38/54 20060101
A61K038/54; A61K 9/00 20060101 A61K009/00; A61K 38/47 20060101
A61K038/47; A61K 38/48 20060101 A61K038/48; A61K 38/46 20060101
A61K038/46; A61K 39/145 20060101 A61K039/145; C12N 7/00 20060101
C12N007/00 |
Claims
1. A method for the treatment or prevention of Influenza in a
mammal or a bird, comprising administering to the mammal or bird a
therapeutically effective amount of a pharmaceutical composition
comprising one or more digestive enzymes.
2. The method of claim 1 wherein the Influenza is Influenza Type
A.
3. The method of claim 2 where the Influenza Type is Subtype
H1N1.
4. The method of claim 1 wherein the one or more digestive enzymes
comprise one or more enzymes selected from the group consisting of
proteases, amylases, celluloses, sucrases, maltases, papain, and
lipases.
5. The method of claim 1 wherein the one or more digestive enzymes
comprise one or more pancreatic enzymes.
6. The method of claim 1 wherein the one or more of the digestive
enzymes comprise avian enzymes or pig enzymes.
7. The method of claim 4 wherein the proteases comprise
chymotrypsin and trypsin.
8. The method of claim 1 wherein the one or more digestive enzymes
are, independently, derived from an animal source, a microbial
source, a plant source, a fungal source, or are synthetically
prepared.
9. The method of claim 1 wherein the mammal is a pig, horse, cow,
dog, cat, monkey, rat, mouse, sheep, goat or human.
10. The method of claim 8 where the animal source is a pig
pancreas.
11. The method of claim 1 wherein the pharmaceutical composition
comprises at least one amylase, a mixture of proteases comprising
chymotrypsin and trypsin, and at least one lipase.
12. The method of claim 1, further comprising vaccinating the
mammal or bird with a TIV or LAIV, or treating the mammal or bird
with an anti-viral medication, or both.
13. The method of claim 1 wherein the pharmaceutical composition
comprises: amylases from about 10,000 to about 60,000 U.S.P,
proteases from about 10,000 to about 70,000 U.S.P, lipases from
about 4,000 to about 30,000 U.S.P., chymotrypsin from about 2 to
about 5 mg, trypsin from about 60 to about 100 mg, papain from
about 3,000 to about 10,000 USP units, and papaya from about 30 to
about 60 mg.
14. The method of claim 1 wherein the pharmaceutical composition
comprises at least one protease and at least one lipase, and
wherein the ratio of total proteases to total lipases (in USP
units) ranges from about 1:1 to about 20:1.
15. The method of claim 11 wherein the ratio of proteases to
lipases ranges from about 4:1 to about 10:1.
16. The method of claim 1 wherein the pharmaceutical composition is
a dosage formulation selected from the group consisting of: pills,
tablets, capsules, microcapsules, mini-capsules, time released
capsules, mini-tabs, sprinkles, and a combination thereof.
17. A method of diagnosing a patient as immune-compromised
comprising: a. obtaining a fecal sample from the patient; b.
determining a level of chymotrypsin present in the fecal sample;
and c. diagnosing the patient as having a compromised immune system
if the determined fecal chymotrypsin level is less than a control
level.
18. The method of claim 17 wherein the determined fecal
chymotrypsin level is less than 8.4 U/gram.
19. The method of claim 17 wherein the determined fecal
chymotrypsin level is less than 4.2 U/gram.
20. The method of claim 17 wherein the level of chymotrypsin
present in the fecal sample is determined using an enzymatic
photospectrometry method.
21. The method of claim 17 further comprising administering to the
patient an effective amount of a pharmaceutical composition
comprising one or more digestive enzymes if the patient is
diagnosed as having a compromised immune system.
22. The method of claim 21 further comprising determining if the
administration of the pharmaceutical composition reduces or
ameliorates the propensity or frequency of contracting bacterial or
viral illnesses or the severity of bacterial or viral
illnesses.
23. The method of claim 22 further comprising comparing the
post-administration measurement of one or more symptoms of the
illness to a pre-administration measurement of the one or more
symptoms of illness.
24. A method of identifying a patient likely to benefit from
administration of a pharmaceutical composition comprising one or
more digestive enzymes comprising: obtaining a fecal sample from
the patient; determining a level of chymotrypsin present in the
fecal sample; and identifying the patient as likely to benefit from
administration of the pharmaceutical composition if the determined
fecal chymotrypsin level is less than a control level and the
patient is diagnosed with a compromised immune system or the
patient is exhibiting one or more symptoms associated with a
bacterial or viral illness.
25. The method of claim 24 further comprising determining if the
patient exhibits one or more symptoms of a compromised immune
system.
26. The method of claim 24 wherein the benefit comprises a
reduction or amelioration of one or more symptoms associated with a
bacterial or viral illness.
27. The method of claim 24 wherein the level of chymotrypsin
present in the fecal sample is determined using an enzymatic
photospectrometry method.
28. The method of claim 24 further comprising administering to the
patient an effective amount of a pharmaceutical composition
comprising one or more digestive enzymes.
29. A pharmaceutical composition comprising one or more digestive
enzymes, wherein the one or more digestive enzymes comprise at
least one lipase and at least one protease, and wherein the ratio
of total proteases to total lipases (in USP units) ranges from
about 1:1 to about 20:1.
30. The pharmaceutical composition of claim 29 wherein the ratio of
total proteases to total lipases ranges from about 4:1 to about
10:1.
31. A pharmaceutical composition comprising at least one amylase, a
mixture of proteases comprising chymotrypsin and trypsin, and at
least one lipase.
32. The pharmaceutical composition of claim 31 wherein the
pharmaceutical composition further comprises papain.
33. The pharmaceutical composition of claim 29 or 31 wherein the
pharmaceutical composition is lipid encapsulated.
34. A method for treating a mammal or bird exhibiting one or more
symptoms of Influenza comprising administering to the mammal or
bird a therapeutically effective amount of a composition comprising
one or more digestive enzymes.
35. The method of claim 34 where the symptoms of influenza are
selected from the group consisting of: fever, headache, tiredness,
cough, sore throat, runny or stuffy nose, body aches, diarrhea and
vomiting, and a combination thereof.
36. The method of claim 34 wherein the mammal is a human or a
pig.
37. The method of claim 34 wherein the preparation is administered
orally via a dosage formulation selected from the group consisting
of: emulsions, liquids, pills, tablets, capsules, microcapsules,
mini-capsules, time released capsules, mini-tabs, powders,
sprinkles, and a combination thereof.
38. A method of preventing infection of an individual with
Influenza or of treating an individual diagnosed with Influenza
comprising: measuring a level of fecal chymotrypsin in a stool
sample of the individual; comparing the level of fecal chymotrypsin
with a normal fecal chymotrypsin level; and administering a
composition comprising one or more digestive enzymes to the
individual if the level of fecal chymotrypsin in the individual is
less than a normal fecal chymotrypsin level.
39. A method for sanitizing or disinfecting a surface to reduce the
amount of influenza virus thereon or to eradicate the influenza
virus thereon, comprising applying to the surface a composition
comprising one or more digestive enzymes.
40. A method for reducing the amount of influenza virus present on
a skin region, tissue, or wound of a mammal or bird comprising
applying to the skin region, tissue, or wound a composition
comprising one or more digestive enzymes.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims priority under 35 U.S.C. .sctn. 119
to U.S. Prov. Appl. Ser. No. 61/253,805, filed Oct. 21, 2009,
incorporated by reference in its entirety herein.
TECHNICAL FIELD
[0002] This disclosure relates to the prevention and treatment of
Influenza, including Influenza A Virus Subtype H1N1, with the use
of a pharmaceutical composition comprising one or more digestive
enzymes. The disclosure further relates to the use of an
individual's fecal chymotrypsin level as an indicator, e.g.,
biomarker, of whether an individual may be immune-compromised
and/or whether an individual may be susceptible to Influenza in
general, and in particular to Influenza A Subtype H1N1, and/or as a
biomarker of whether an individual will benefit from administration
of a described pharmaceutical composition. This disclosure also
relates to the use of compositions comprising one or more digestive
enzymes as antiseptics, detergents, sanitizers, and disinfectants,
e.g., as viricidal and/or viristatic compositions, to treat
surfaces and thus to prevent and/or reduce the spread of Influenza
infections.
BACKGROUND
[0003] Digestive enzymes are enzymes that can break down one or
more components of foods, e.g., carbohydrates, lipids/fats,
proteins, cellulose, nucleic acids, etc., and thereby assist in
digestion and uptake of nutrients. Certain 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. Other digestive
enzymes are produced by plants, fungi, or microorganisms (e.g.,
papain, bromelain).
[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
such as enzymes have been described, see, e.g., U.S. RE40,059;
6,835,397; 6,797,291; 6,616,954; 6,312,741; 6,251,478; 6,153,236;
6,013,286, and 5,190,775, and Ser. No. 12/386,051, all of which are
incorporated by reference in their entirety herein.
[0006] Influenza A (H1N1) virus is a subtype of influenza virus A
and the most common cause of influenza (flu) in humans. Some
strains of H1N1 are endemic in humans and cause a small fraction of
all influenza-like illness and a large fraction of all seasonal
influenza. H1N1 strains caused roughly half of all human flu
infections in 2006. Other strains of H1N1 are endemic in pigs
(swine influenza) and in birds (avian influenza).
[0007] In June 2009, WHO declared that flu due to a new strain of
swine-origin H1N1 was responsible for the 2009 flu pandemic. This
strain is commonly called "swine flu" by the public media.
[0008] Influenza A virus strains are categorized according to two
proteins found on the surface of the virus: hemagglutinin (H) and
neuraminidase (N). All influenza A viruses contain hemagglutinin
and neuraminidase, but the structure of these proteins differ from
strain to strain due to rapid genetic mutation in the viral genome.
Influenza A virus strains are assigned an H number and an N number
based on which forms of these two proteins the strain contains.
There are 16 H and 9 N subtypes known in birds, but only H 1, 2 and
3, and N 1 and 2 are commonly found in humans.
[0009] The Spanish flu, also known as La Gripe Espanola, or La
Pesadilla, was an unusually severe and deadly strain of avian
influenza, a viral infectious disease, that killed some 50 million
to 100 million people worldwide over about a year in 1918 and 1919.
It is thought to be one of the most deadly pandemics in human
history. Tt was caused by the H1N1 type of influenza virus. It is
postulated that the Spanish flu caused an unusual number of deaths
because it may have caused a cytokine storm in the body. The recent
epidemic of bird flu, also an Influenza A virus, had a similar
effect. The Spanish flu virus infected lung cells, leading to
overstimulation of the immune system via release of cytokines into
the lung tissue. This leads to extensive leukocyte migration
towards the lungs, causing destruction of lung tissue and secretion
of liquid into the organ, making it difficult for the patient to
breathe. In contrast to other pandemics, which mostly kill the old
and the very young, the 1918 pandemic killed unusual numbers of
young adults, which may have been due to their healthy immune
systems being able to mount a very strong and damaging response to
the infection.
[0010] The more recent Russian flu was a 1977-1978 flu epidemic
caused by strain Influenza A/USSR/90/77 (H1N1). It infected mostly
children and young adults under 23 because a similar strain was
prevalent in 1947-57, causing most adults to have substantial
immunity. Some have called it a flu pandemic, but because it only
affected the young it is not considered a true pandemic. The virus
was included in the 1978-1979 influenza vaccine.
[0011] In the 2009 flu pandemic, the virus isolated from patients
in the United States was found to be made up of genetic elements
from four different flu viruses--North American Mexican influenza,
North American avian influenza, human influenza, and swine
influenza virus typically found in Asia and Europe. This strain
appears to be a result of reassortment of human influenza and swine
influenza viruses, in all four different strains of subtype
H1N1.
[0012] Preliminary genetic characterization found that the
hemagglutinin (HA) gene was similar to that of swine flu viruses
present in U.S. pigs since 1999, but the neuraminidase (NA) and
matrix protein (M) genes resembled versions present in European
swine flu isolates. The six genes from American swine flu are
themselves mixtures of swine flu, bird flu, and human flu viruses.
While viruses with this genetic makeup had not previously been
found to be circulating in humans or pigs, there is no formal
national surveillance system to determine what viruses are
circulating in pigs in the U.S.
[0013] On Jun. 11, 2009, the WHO declared an H1N1 pandemic, moving
the alert level to phase 6, marking the first global pandemic since
1968.
[0014] Communicable diseases are currently the leading cause of
preventable deaths worldwide, disproportionately affecting
resource-poor settings. Pandemic influenzas add to already
unacceptable levels of morbidity and mortality from diarrhea,
malaria, pneumonia, malnutrition, HIV/AIDS and tuberculosis, in
addition to causing high maternal and neonatal death rates. A few
key conditions cause 90% of deaths from communicable diseases:
pneumonia (3.9 million deaths per year); diarrhoeal diseases (1.8
million); and malaria (1.2 million). Malnutrition is a significant
contributing factor to this mortality. During a pandemic, these
illnesses are likely to increase in resource-poor settings where
chronically strained health systems would face even higher patient
volumes, severe resource constraints, and absenteeism of critical
staff.
[0015] The current prophylactic means for preventing the flu is by
Injectable Inactivated Vaccine or Nasal Spray Flu Vaccine. Commonly
called the "flu shot", the Injectable Inactivated Vaccine method
employs an inactivated vaccine (containing killed virus) that is
given with a needle, usually in the arm. The flu shot is approved
for use in people older than 6 months, including healthy people and
people with chronic medical conditions. Alternately, the
nasal-spray flu vaccine--a vaccine made with live, weakened flu
viruses that do not cause the flu (sometimes called LAIV for "live
attenuated influenza vaccine" or FluMist.RTM.). LAIV (FluMist.RTM.)
is approved for use in healthy people 2-49 years of age who are not
pregnant.
[0016] Typically each vaccine contains three influenza viruses-one
A (H3N2) virus, one A (H1N1) virus, and one B virus. The viruses in
the vaccine change each year based on international surveillance
and scientists' estimations about which types and strains of
viruses will circulate in a given year. About 2 weeks after
vaccination, antibodies that provide protection against influenza
virus infection develop in the body.
[0017] Annual influenza vaccination is the most effective method
for preventing influenza virus infection and its complications.
Influenza vaccine can be administered to any person aged >6
months (who does not have contraindications to vaccination) to
reduce the likelihood of becoming ill with influenza or of
transmitting influenza to others. Trivalent inactivated influenza
vaccine (TIV) can be used for any person aged >6 months,
including those with high-risk conditions. Live, attenuated
influenza vaccine (LAW) may be used for healthy, nonpregnant
persons aged 2-49 years. If vaccine supply is limited, priority for
vaccination is typically assigned to persons in specific groups and
of specific ages who are, or are contacts of, persons at higher
risk for influenza complications. Because the safety or
effectiveness of LAIV has not been established in persons with
underlying medical conditions that confer a higher risk for
influenza complications, these persons should only be vaccinated
with TIV. Influenza viruses undergo frequent antigenic change
(i.e., antigenic drift), and persons recommended for vaccination
must receive an annual vaccination against the influenza viruses
forecasted to be in circulation. Although vaccination coverage has
increased in recent years for many groups targeted for routine
vaccination, coverage remains low among most of these groups.
[0018] Antiviral medications are an adjunct to vaccination and are
effective when administered as treatment and when used for
chemoprophylaxis after an exposure to influenza virus. Oseltamivir
and zanamivir are the only antiviral medications recommended for
use in the United States. Amantadine or rimantidine should not be
used for the treatment or prevention of influenza in the United
States until evidence of susceptibility to these antiviral
medications has been reestablished among circulating influenza A
viruses.
[0019] The efficacy (i.e., prevention of illness among vaccinated
persons in controlled trials) and effectiveness (i.e., prevention
of illness in vaccinated populations) of influenza vaccines depend
in part on the age and immunocompetence of the vaccine recipient,
the degree of similarity between the viruses in the vaccine and
those in circulation, and the outcome being measured. Influenza
vaccine efficacy and effectiveness studies have used multiple
possible outcome measures, including the prevention of medically
attended acute respiratory illness (MAARI), prevention of
laboratory-confirmed influenza virus illness, prevention of
influenza or pneumonia-associated hospitalizations or deaths, or
prevention of seroconversion to circulating influenza virus
strains.
[0020] Efficacy or effectiveness for more specific outcomes such as
laboratory-confirmed influenza typically will be higher than for
less specific outcomes such as MAARI because the causes of MAARI
include infections with other pathogens that influenza vaccination
would not be expected to prevent. Observational studies that
compare less-specific outcomes among vaccinated populations to
those among unvaccinated populations are subject to biases that are
difficult to control for during analyses. For example, an
observational study that determines that influenza vaccination
reduces overall mortality might be biased if healthier persons in
the study are more likely to be vaccinated. Randomized controlled
trials that measure laboratory-confirmed influenza virus infections
as the outcome are the most persuasive evidence of vaccine
efficacy, but such trials cannot be conducted ethically among
groups recommended to receive vaccine annually.
[0021] Both LAIV and TIV contain strains of influenza viruses that
are antigenically equivalent to the annually recommended strains:
one influenza A (H3N2) virus, one influenza A (H1N1) virus, and one
influenza B virus. Each year, one or more virus strains in the
vaccine might be changed on the basis of global surveillance for
influenza viruses and the emergence and spread of new strains. All
three vaccine virus strains were changed for the recommended
vaccine for the 2008-09 influenza season, compared with the 2007-08
season.
[0022] During the preparation of TIV, the vaccine viruses are made
noninfectious (i.e., inactivated or killed). Only subvirion and
purified surface antigen preparations of T1V (often referred to as
"split" and subunit vaccines, respectively) are available in the
United States. TIV contains killed viruses and thus cannot cause
influenza. LAIV contains live, attenuated viruses that have the
potential to cause mild signs or symptoms such as runny nose, nasal
congestion, fever or sore throat. LAIV is administered intranasally
by sprayer, whereas TIV is administered intramuscularly by
injection. LAW is licensed for use among nonpregnant persons aged
2-49 years; safety has not been established in persons with
underlying medical conditions that confer a higher risk of
influenza complications. TIV is licensed for use among persons aged
>6 months, including those who are healthy and those with
chronic medical conditions. LAW is generally regarded as more
efficacious and effective than TIV.
[0023] In many populations, there remains a need for alternatives
to TIV and LAW, and a need for adjunctive prophylactic or
therapeutic regimens for the prevention and/or treatment of
Influenza.
[0024] Pregnant Women and Neonates--
[0025] Pregnant women have protective levels of anti-influenza
antibodies after vaccination. Passive transfer of anti-influenza
antibodies that might provide protection from vaccinated women to
neonates has been reported. A retrospective, clinic-based study
conducted during 1998-2003 documented a non-significant trend
towards fewer episodes of MAARI during one influenza season among
vaccinated pregnant women compared with unvaccinated pregnant women
and substantially fewer episodes of MAARI during the peak influenza
season. However, a retrospective study conducted during 1997-2002
that used clinical records data did not indicate a reduction in ILI
among vaccinated pregnant women or their infants. In another study
conducted during 1995-2001, medical visits for respiratory illness
among the infants were not substantially reduced. However, studies
of influenza vaccine effectiveness among pregnant women have not
included specific outcomes such as laboratory-confirmed influenza
in women or their infants.
[0026] Elderly Population--
[0027] Adults aged >65 years typically have a diminished immune
response to influenza vaccination compared with young healthy
adults, suggesting that immunity might be of shorter duration
(although still extending through one influenza season). However, a
review of the published literature concluded that no clear evidence
existed that immunity declined more rapidly in the elderly.
Infections among the vaccinated elderly might be associated with an
age-related reduction in ability to respond to vaccination rather
than reduced duration of immunity. The only randomized controlled
trial among community-dwelling persons aged >60 years reported a
vaccine efficacy of 58% against influenza respiratory illness
during a season when the vaccine strains were considered to be
well-matched to circulating strains, but indicated that efficacy
was lower among those aged >70 years. Influenza vaccine
effectiveness in preventing MAARI among the elderly in nursing
homes has been estimated at 20%-40%, and reported outbreaks among
well-vaccinated nursing home populations have suggested that
vaccination might not have any significant effectiveness when
circulating strains are drifted from vaccine strains. In contrast,
some studies have indicated that vaccination can be up to 80%
effective in preventing influenza-related death. Among elderly
persons not living in nursing homes or similar chronic-care
facilities, influenza vaccine is 27%-70% effective in preventing
hospitalization for pneumonia and influenza. Influenza vaccination
reduces the frequency of secondary complications and reduces the
risk for influenza-related hospitalization and death among
community-dwelling adults aged >65 years with and without
high-risk medical conditions (e.g., heart disease and diabetes).
However, studies demonstrating large reductions in hospitalizations
and deaths among the vaccinated elderly have been conducted using
medical record databases and have not measured reductions in
laboratory-confirmed influenza illness. These studies have been
challenged because of concerns that they have not adequately
controlled for differences in the propensity for healthier persons
to be more likely than less healthy persons to receive
vaccination.
[0028] HIV Compromised Individuals--
[0029] TIV produces adequate antibody concentrations against
influenza among vaccinated HIV-infected persons who have minimal
AIDS-related symptoms and normal or near-normal CD4+T-lymphocyte
cell counts. Among persons who have advanced HIV disease and low
CD4+T-lymphocyte cell counts, TIV might not induce protective
antibody titers; a second dose of vaccine does not improve the
immune response in these persons. A randomized, placebo-controlled
trial determined that TIV was highly effective in preventing
symptomatic, laboratory-confirmed influenza virus infection among
HIV-infected persons with a mean of 400 CD4+T-lymphocyte cells/mm3;
however, a limited number of persons with CD4+ T-lymphocyte cell
counts of <200 were included in that study. A nonrandomized
study of HIV-infected persons determined that influenza vaccination
was most effective among persons with >100 CD4+ cells and among
those with <30,000 viral copies of HIV type-1/mL.
[0030] Transplant Recipients--
[0031] On the basis of certain small studies, immunogenicity for
persons with solid organ transplants varies according to transplant
type. Among persons with kidney or heart transplants, the
proportion that developed seroprotective antibody concentrations
was similar or slightly reduced compared with healthy persons.
However, a study among persons with liver transplants indicated
reduced immunologic responses to influenza vaccination, especially
if vaccination occurred within the 4 months after the transplant
procedure.
[0032] Other Medical Conditions--
[0033] persons with underlying medical conditions including asthma,
reactive airways disease, or other chronic disorders of the
pulmonary or cardiovascular systems; metabolic diseases such as
diabetes, renal dysfunction, and hemoglobinopathies; or known or
suspected immunodeficiency diseases or immunosuppressed states
should not be vaccinated with LAIV. In addition children or
adolescents receiving aspirin or other salicylates should not be
vaccinated with a LAIV because of the association of Reye syndrome
and salicylates with wild-type influenza virus infection.
Individuals with acute febrile illness should not be vaccinated
with TIV or LAW.
[0034] Pediatric Chronic Medical Conditions--
[0035] Among children with high-risk medical conditions, one study
of 52 children aged 6 months-3 years reported fever among 27% and
irritability and insomnia among 25% (113); and a study among 33
children aged 6-18 months reported that one child had irritability
and one had a fever and seizure after vaccination. No placebo
comparison group was used in these studies.
[0036] Hypersensitivity and Allergic Reactions--
[0037] Immediate and presumably allergic reactions (e.g., hives,
angioedema, allergic asthma, and systemic anaphylaxis) occur rarely
after influenza vaccination. These reactions probably result from
hypersensitivity to certain vaccine components; the majority of
reactions probably are caused by residual egg protein. Although
influenza vaccines contain only a limited quantity of egg protein,
this protein can induce immediate hypersensitivity reactions among
persons who have severe egg allergy. Manufacturers use a variety of
different compounds to inactivate influenza viruses and add
antibiotics to prevent bacterial contamination. Persons who have
experienced hives or swelling of the lips or tongue, or who have
experienced acute respiratory distress or who collapse after eating
eggs, must consult a physician for appropriate evaluation to help
determine if vaccine should be administered. Persons who have
documented immunoglobulin E (IgE)-mediated hypersensitivity to
eggs, including those who have had occupational asthma related to
egg exposure or other allergic responses to egg protein, also might
be at increased risk for allergic reactions to influenza vaccine,
and consultation with a physician before vaccination must be
considered. Hypersensitivity reactions to other vaccine components
can occur but are rare. Although exposure to vaccines containing
thimerosal can lead to hypersensitivity, the majority of patients
do not have reactions to thimerosal when it is administered as a
component of vaccines, even when patch or intradermal tests for
thimerosal indicate hypersensitivity. When reported,
hypersensitivity to thimerosal typically has consisted of local
delayed hypersensitivity reactions.
[0038] Guillain-Barre Syndrome--
[0039] The annual incidence of Guillain-Barre Syndrome (GBS) is
10-20 cases per 1 million adults. Substantial evidence exists that
multiple infectious illnesses, most notably Campylobacter jejuni
gastrointestinal infections and upper respiratory tract infections,
are associated with GBS. The 1976 swine influenza vaccine was
associated with an increased frequency of GBS, estimated at one
additional case of GBS per 100,000 persons vaccinated. The risk for
influenza vaccine-associated GBS was higher among persons aged
>25 years than among persons aged <25 years. However,
obtaining strong epidemiologic evidence for a possible small
increase in risk for a rare condition with multiple causes is
difficult, and no evidence exists for a consistent causal relation
between subsequent vaccines prepared from other influenza viruses
and GBS.
[0040] None of the studies conducted using influenza vaccines other
than the 1976 swine influenza vaccine have demonstrated a
substantial increase in GBS associated with influenza vaccines.
During three of four influenza seasons studied during 1977-1991,
the overall relative risk estimates for GBS after influenza
vaccination were not statistically significant in any of these
studies. However, in a study of the 1992-93 and 1993-94 seasons,
the overall relative risk for GBS was 1.7 (CI=1.0-2.8; p=0.04)
during the 6 weeks after vaccination, representing approximately
one additional case of GBS per 1 million persons vaccinated; the
combined number of GBS cases peaked 2 weeks after vaccination.
Results of a study that examined health-care data from Ontario,
Canada, during 1992-2004 demonstrated a small but statistically
significant temporal association between receiving influenza
vaccination and subsequent hospital admission for GBS. However, no
increase in cases of GBS at the population level was reported after
introduction of a mass public influenza vaccination program in
Ontario beginning in 2000. Data from VAERS have documented
decreased reporting of GBS occurring after vaccination across age
groups over time, despite overall increased reporting of other,
non-GBS conditions occurring after administration of influenza
vaccine. Cases of GBS after influenza virus infection have been
reported, but no other epidemiologic studies have documented such
an association. Recently published data from the United Kingdom's
General Practice Research Database (GPRD) found influenza vaccine
to be protective against GBS, although it is unclear if this was
associated with protection against influenza or confounding because
of a "healthy vaccine" (e.g., healthier persons might be more
likely to be vaccinated and are lower risk for GBS). A separate
GPRD analysis found no association between vaccination and GBS over
a 9 year period; only three cases of GBS occurred within 6 weeks
after influenza vaccine.
[0041] It is not known if GBS is a side effect of influenza
vaccines other than 1976 swine influenza vaccine; the estimated
risk for GBS (on the basis of the few studies that have
demonstrated an association between vaccination and GBS) is low
(i.e., approximately one additional case per 1 million persons
vaccinated). It has been deemed by the CDC and others that the
potential benefits of influenza vaccination in preventing serious
illness, hospitalization, and death substantially outweigh these
estimates of risk for vaccine-associated GBS. No evidence indicates
that the case fatality ratio for GBS differs among vaccinated
persons and those not vaccinated
[0042] The incidence of GBS among the general population is low,
but persons with a history of GBS have a substantially greater
likelihood of subsequently experiencing GBS when injected with TIV
influenza vaccine than persons without such a history. Thus, the
likelihood of coincidentally experiencing GBS after influenza
vaccination is expected to be greater among persons with a history
of GBS than among persons with no history of this syndrome. Whether
influenza vaccination specifically might increase the risk for
recurrence of GBS is unknown. However, avoiding vaccinating persons
who are not at high risk for severe influenza complications and who
are known to have experienced GBS within 6 weeks after a previous
influenza vaccination is often taken as a prudent as a precaution.
As an alternative, physicians use influenza antiviral
chemoprophylaxis for these persons. Although data are limited, the
established benefits of influenza vaccination might outweigh the
risks for many persons who have a history of GBS and who are also
at high risk for severe complications from influenza.
[0043] Viral Shedding--
[0044] Available data indicates that both children and adults
vaccinated with LAIV can shed vaccine viruses after vaccination,
although in lower amounts than occur typically with shedding of
wild-type influenza viruses. In rare instances, shed vaccine
viruses can be transmitted from vaccine recipients to unvaccinated
persons. However, serious illnesses have not been reported among
unvaccinated persons who have been infected inadvertently with
vaccine viruses.
[0045] One study of children aged 8-36 months in a child care
center assessed transmissibility of vaccine viruses from 98
vaccinated to 99 unvaccinated subjects; 80% of vaccine recipients
shed one or more virus strains (mean duration: 7.6 days). One
influenza type B vaccine strain isolate was recovered from a
placebo recipient and was confirmed to be vaccine-type virus. The
type B isolate retained the cold-adapted, temperature-sensitive,
attenuated phenotype, and it possessed the same genetic sequence as
a virus shed from a vaccine recipient who was in the same play
group. The placebo recipient from whom the influenza type B vaccine
strain was isolated had symptoms of a mild upper respiratory
illness but did not experience any serious clinical events. The
estimated probability of acquiring vaccine virus after close
contact with a single LAIV recipient in this child care population
was 0.6%-2.4%.
[0046] Studies assessing whether vaccine viruses are shed have been
based on viral cultures or PCR detection of vaccine viruses in
nasal aspirates from persons who have received LAIV. One study of
20 healthy vaccinated adults aged 18-49 years demonstrated that the
majority of shedding occurred within the first 3 days after
vaccination, although the vaccine virus was detected in one subject
on day 7 after vaccine receipt. Duration or type of symptoms
associated with receipt of LAIV did not correlate with detection of
vaccine viruses in nasal aspirates. Another study in 14 healthy
adults aged 18-49 years indicated that 50% of these adults had
viral antigen detected by direct immunofluorescence or rapid
antigen tests within 7 days of vaccination. The majority of samples
with detectable virus were collected on day 2 or 3. Vaccine strain
virus was detected from nasal secretions in one (2%) of 57
HIV-infected adults who received LAIV, none of 54 HIV-negative
participants (256), and three (13%) of 23 HIV-infected children
compared with seven (28%) of 25 children who were not HIV-infected.
No participants in these studies had detectable virus beyond 10
days after receipt of LAIV. The possibility of person-to-person
transmission of vaccine viruses was not assessed in these
studies.
[0047] LAIV Side Effects--
[0048] In a subset of healthy children aged 60-71 months from one
clinical trial (233), certain signs and symptoms were reported more
often after the first dose among LAIV recipients (n=214) than among
placebo recipients (n=95), including runny nose (48% and 44%,
respectively); headache (18% and 12%, respectively); vomiting (5%
and 3%, respectively); and myalgias (6% and 4%, respectively).
However, these differences were not statistically significant. In
other trials, signs and symptoms reported after LAIV administration
have included runny nose or nasal congestion (20%-75%), headache
(2%-46%), fever (0-26%), vomiting (3%-13%), abdominal pain (2%),
and myalgias (0-21%). These symptoms were associated more often
with the first dose and were self-limited.
[0049] In a randomized trial published in 2007, LAIV and TIV were
compared among children aged 6-59 months. Children with medically
diagnosed or treated wheezing within 42 days before enrollment, or
a history of severe asthma, were excluded from this study. Among
children aged 24-59 months who received LAIV, the rate of medically
significant wheezing, using a pre-specified definition, was not
greater compared with those who received TIV; wheezing was observed
more frequently among younger LAW recipients in this study. In a
previous randomized placebo-controlled safety trial among children
aged 12 months-17 years without a history of asthma by parental
report, an elevated risk for asthma events (RR=4.06, CI=1.29-17.86)
was documented among 728 children aged 18-35 months who received
LAIV. Of the 16 children with asthma-related events in this study,
seven had a history of asthma on the basis of subsequent medical
record review. None required hospitalization, and elevated risks
for asthma were not observed in other age groups.
[0050] Among adults aged 19-49, runny nose or nasal congestion
(28%-78%), headache (16%-44%), and sore throat (15%-27%) have been
reported more often among vaccine recipients than placebo
recipients. In one clinical trial among a subset of healthy adults
aged 18-49 years, signs and symptoms reported more frequently among
LAW recipients (n=2,548) than placebo recipients (n=1,290) within 7
days after each dose included cough (14% and 11%, respectively);
runny nose (45% and 27%, respectively); sore throat (28% and 17%,
respectively); chills (9% and 6%, respectively); and
tiredness/weakness (26% and 22%, respectively).
[0051] There are additional reasons why it would be useful to have
alternative or adjunctive prophylactic and/or therapeutic options
for the prevention and/or treatment of Influenza, as discussed
below.
[0052] Challenging Prediction of Virus Strains--
[0053] Manufacturing trivalent influenza virus vaccines is a
challenging process that takes 6-8 months to complete. This
manufacturing timeframe requires that influenza vaccine strains for
influenza vaccines used in the United States must be selected in
February of each year by the FDA to allow time for manufacturers to
prepare vaccines for the next influenza season. Vaccine strain
selections are based on global viral surveillance data that is used
to identify trends in antigenic changes among circulating influenza
viruses and the availability of suitable vaccine virus
candidates.
[0054] Vaccination can provide reduced but substantial
cross-protection against drifted strains in some seasons, including
reductions in severe outcomes such as hospitalization. Usually one
or more circulating viruses with antigenic changes compared with
the vaccine strains are identified in each influenza season.
However, assessment of the clinical effectiveness of influenza
vaccines cannot be determined solely by laboratory evaluation of
the degree of antigenic match between vaccine and circulating
strains. In some influenza seasons, circulating influenza viruses
with significant antigenic differences predominate and, compared
with seasons when vaccine and circulating strains are well-matched,
reductions in vaccine effectiveness are sometimes observed.
However, even during years when vaccine strains were not
antigenically well matched to circulating strains, substantial
protection has been observed against severe outcomes, presumably
because of vaccine-induced cross-reacting antibodies. For example,
in one study conducted during an influenza season (2003-04) when
the predominant circulating strain was an influenza A (H3N2) virus
that was antigenically different from that season's vaccine strain,
effectiveness among persons aged 50-64 years against
laboratory-confirmed influenza illness was 60% among healthy
persons and 48% among persons with medical conditions that increase
risk for influenza complications. An interim, within-season
analysis during the 2007-08 influenza season indicated that vaccine
effectiveness was 44% overall, 54% among healthy persons aged 5-49
years, and 58% against influenza A, despite the finding that
viruses circulating in the study area were predominately a drifted
influenza A H3N2 and a influenza B strain from a different lineage
compared with vaccine strains. Among children, both TIV and LAW
provide protection against infection even in seasons when vaccines
and circulating strains are not well matched. Vaccine effectiveness
against ILI was 49%-69% in two observational studies, and 49%
against medically attended, laboratory-confirmed influenza in a
case-control study conducted among young children during the
2003-04 influenza season, when a drifted influenza A H3N2 strain
predominated, based on viral surveillance data. However, the FDA
admits that continued improvements in collecting representative
circulating viruses and use surveillance data to forecast antigenic
drift are needed. Shortening manufacturing time to increase the
time to identify good vaccine candidate strains from among the most
recent circulating strains also is also important. Data from
multiple seasons and collected in a consistent manner are needed to
better understand vaccine effectiveness during seasons when
circulating and vaccine virus strains are not well-matched.
[0055] Vaccine Coverage--
[0056] vaccination coverage of the population is influenced by a
multitude of factors including vaccine supply delays and shortages,
changes in influenza vaccination recommendations and target groups
for vaccination, reimbursement rates for vaccine and vaccine
administration, and other factors related to vaccination coverage
among adults and children. Production issues coupled with the
requirement to forecast appropriate influenza types and antigens
have caused severe shortfalls in vaccine availability, for example
in the years 2004-05 an American company, Chiron, had their
operating license suspended by British officials following problems
at their manufacturing plant in Liverpool, England. Due to
contamination in a batch of vaccines intended for American market
they were unable to supply their flu vaccine, Fluvirin. Fluvirin
made up approximately 50% of America's expected demand for the
winter flu season.
[0057] Because of the inherent risk factors and the reluctance of
individuals to accept those risks, many at risk groups have very
low vaccination coverage. For example vaccine coverage among
pregnant women has not increased significantly during the preceding
decade. Only 12% and 13% of pregnant women participating in the
2006 and 2007 NHIS reported vaccination during the 2005-06 and
2006-07 seasons, respectively, excluding pregnant women who
reported diabetes, heart disease, lung disease, and other selected
high-risk conditions. In a study of influenza vaccine acceptance by
pregnant women, 71% of those who were offered the vaccine chose to
be vaccinated. However, a 1999 survey of obstetricians and
gynecologists determined that only 39% administered influenza
vaccine to obstetric patients in their practices, although 86%
agreed that pregnant women's risk for influenza-related morbidity
and mortality increases during the last two trimesters.
[0058] Drug Resistance--
[0059] Viral neuraminidase is an enzyme on the surface of influenza
viruses that enables the virus to be released from the host cell.
Drugs that inhibit neuraminidase are often used to treat influenza.
Neuraminidase has been targeted in structure-based enzyme inhibitor
design programmes that have resulted in the production of two
drugs, zanamivir (Relenza) and oseltamivir (Tamiflu).
Administration of neuraminidase inhibitors is a treatment that
limits the severity and spread of viral infections. Neuraminidase
inhibitors are useful for combating influenza infection: zanamivir,
administered by inhalation; oseltamivir, administered orally; and
under research is peramivir administered parenterally, that is
through intravenous or intramuscular injection.
[0060] On Feb. 27, 2005, a 14-year-old Vietnamese girl was
documented to be carrying an H5N1 influenza virus strain that was
resistant to the drug oseltamivir. The drug is used to treat
patients that have contracted influenza. However, the Vietnamese
girl who had received a prophylaxis dose (75 mg once a day) was
found to be non-responsive to the medication. In growing fears of a
global avian flu pandemic, scientists began to look for a cause of
resistance to the Tamiflu medication. The cause was determined to
be a histidine-to-tryosine (amino acid) substitution at position
274 in its neuraminidase protein.
SUMMARY
[0061] This disclosure relates to the prevention and/or treatment
of Influenza, including Influenza A Virus Subtype H1N1, with the
use of a pharmaceutical composition comprising one or more
digestive enzymes, including pancreatic or other digestive-track
enzymes (e.g., porcine pancreatic enzymes or bird-derived digestive
track enzymes) or plant-, fungal-, or microorganism-derived
enzymes, that break down components of food. As used herein, a
pharmaceutical composition can be used for human or veterinary
indications. Accordingly, the pharmaceutical compositions can be
useful for prophylactic and/or therapeutic treatment of human or
other mammalian populations (e.g., pig, horse, cow, sheep, goat,
monkey, rat, mouse, cat, dog) or of bird populations (e.g., duck,
goose, chicken, turkey). The disclosure further relates to the use
of an individual's (e.g., a human's) fecal chymotrypsin level as an
indicator, e.g., biomarker, of whether an individual may be
immune-compromised and/or whether an individual may be susceptible
to Influenza in general, and in particular to Influenza A Subtype
H1N1, and/or as a biomarker of whether an individual will benefit
from administration of a described pharmaceutical composition,
e.g., to prevent and/or treat Influenza. The compositions can be
used on their own, and/or as adjuncts to vaccination or other
anti-viral regimens, and/or with other therapeutic or anti-viral
agents post-infection to treat Influenza.
[0062] The present disclosure also relates to enzyme delivery
systems comprising digestive enzyme preparations, which are useful
in the prophylaxis and treatment of influenza in general and more
particularly useful for the prevention and treatment of Influenza A
Virus, Subtype H1N1, including H1N1 variants swine influenza
(endemic in pigs) and avian influenza (endemic in birds). In some
embodiments, the digestive enzyme preparations of this disclosure
permit controlled delivery of enzymes having increased stability
and enhanced administration properties for the prevention of
influenza infection and treatment of influenza symptoms.
[0063] In virus classification the influenza virus is an RNA virus
of three of the five genera of the family Orthomyxoviridae:
Influenzavirus A, Influenzavirus B, Influenzavirus C. These viruses
are only distantly related to the human parainfluenza viruses,
which are RNA viruses belonging to the paramyxovirus family that
are a common cause of respiratory infections in children such as
croup, but can also cause a disease similar to influenza in
adults.
[0064] Influenza viruses A, B and C are very similar in overall
structure. The virus particle is 80-120 nanometres in diameter and
usually roughly spherical, although filamentous forms can occur.
These filamentous forms are more common in influenza C, which can
form cordlike structures up to 500 micrometres long on the surfaces
of infected cells. However, despite these varied shapes, the viral
particles of all influenza viruses are similar in composition.
These are made of a viral envelope containing two main types of
glycoproteins, wrapped around a central core. The central core
contains the viral RNA genome and other viral proteins that package
and protect this RNA. Unusually for a virus, its genome is not a
single piece of nucleic acid; instead, it contains seven or eight
pieces of segmented negative-sense RNA, each piece of RNA contains
either one or two genes. For example, the influenza A genome
contains 11 genes on eight pieces of RNA, encoding for 11 proteins:
hemagglutinin (HA), neuraminidase (NA), nucleoprotein (NP), M1, M2,
NS1, NS2(NEP), PA, PB1, PB1-F2 and PB2.
[0065] Hemagglutinin (HA) and neuraminidase (NA) are the two large
glycoproteins on the outside of the viral particles. HA is a lectin
that mediates binding of the virus to target cells and entry of the
viral genome into the target cell, while NA is involved in the
release of progeny virus from infected cells, by cleaving sugars
that bind the mature viral particles. Thus, these proteins are
targets for antiviral drugs. Furthermore, they are antigens to
which antibodies can be raised. Influenza A viruses are classified
into subtypes based on antibody responses to HA and NA. These
different types of HA and NA form the basis of the H and N
distinctions in, for example, H5N1. There are 16 H and 9 N subtypes
known, but only H 1, 2 and 3, and N 1 and 2 are commonly found in
humans.
[0066] There are at least 16 different HA antigens. These subtypes
are labeled H1 through H16. The last, H16, was discovered only
recently on influenza A viruses isolated from black-headed gulls
from Sweden and Norway. The first three hemagglutinins, H1, H2, and
H3, are found in human influenza viruses.
[0067] A highly pathogenic avian flu virus of H5N1 type has been
found to infect humans at a low rate. It has been reported that
single amino acid changes in this avian virus strain's type H5
hemagglutinin have been found in human patients that "can
significantly alter receptor specificity of avian H5N1 viruses,
providing them with an ability to bind to receptors optimal for
human influenza viruses". This finding seems to explain how an H5N1
virus that normally does not infect humans can mutate and become
able to efficiently infect human cells. The hemagglutinin of the
H5N1 virus has been associated with the high pathogenicity of this
flu virus strain, apparently due to its ease of conversion to an
active form by proteolysis.
[0068] HA has two primary functions: allowing the recognition of
target vertebrate cells, accomplished through the binding of these
cells' sialic acid-containing receptors, and allowing the entry of
the viral genome into the target cells by causing the fusion of
host endosomal membrane with the viral membrane.
[0069] HA binds to the monosaccharide sialic acid which is present
on the surface of its target cells. This causes the viral particles
to stick to the cell's surface. The cell membrane then engulfs the
virus and the portion of the membrane that encloses it pinches off
to form a new membrane-bound compartment within the cell called an
endosome, which contains the engulfed virus. The cell then attempts
to begin digesting the contents of the endosome by acidifying its
interior and transforming it into a lysosome. However, as soon as
the pH within the endosome drops to about 6.0, the original folded
structure of the HA molecule becomes unstable, causing it to
partially unfold, and releasing a very hydrophobic portion of its
peptide chain that was previously hidden within the protein. This
so-called "fusion peptide" acts like a molecular grappling hook by
inserting itself into the endosomal membrane and locking on. Then,
when the rest of the HA molecule refolds into a new structure
(which is more stable at the lower pH), it "retracts the grappling
hook" and pulls the endosomal membrane right up next to the virus
particle's own membrane, causing the two to fuse together. Once
this has happened, the contents of the virus, including its RNA
genome, are free to pour out into the cell's cytoplasm.
[0070] HA is a homotrimeric integral membrane glycoprotein. It is
shaped like a cylinder, and is approximately 13.5 nanometres long.
The three identical monomers that constitute HA are constructed
into a central a helix coil; three spherical heads contain the
sialic acid binding sites. HA monomers are synthesized as
precursors that are then glycosylated and cleaved into two smaller
polypeptides: the HA1 and HA2 subunits. Each HA monomer consists of
a long, helical chain anchored in the membrane by HA2 and topped by
a large HA' globule.
[0071] Viral neuraminidase is an enzyme on the surface of influenza
viruses that enables the virus to be released from the host cell.
Drugs that inhibit neuraminidase are often used to treat influenza.
When influenza virus reproduces, it moves to the cell surface with
a hemagglutinin molecule on the surface of the virus bound to a
sialic acid receptor on the surface of the cell. In order for the
virus to be released free from the cell, neuraminidase must break
apart (cleave) the sialic acid receptor. In some viruses, a
hemagglutinin-neuraminidase protein combines the neuraminidase and
hemagglutinin functions in a single protein.
[0072] The viral neuraminidase enzyme helps viruses to be released
from a host cell.
[0073] Influenza virus membranes contain two glycoproteins:
haemagglutinin and neuraminidase. While the hemagglutinin on the
surface of the virion is needed for infection, its presence
inhibits release of the particle after budding. It also mediates
cell-surface sialic acid receptor binding to initiate virus
infection. Viral neuraminidase cleaves terminal neuraminic acid
(also called sialic acid) residues from glycan structures on the
surface of the infected cell. This promotes the release of progeny
viruses and the spread of the virus from the host cell to
uninfected surrounding cells. Neuraminidase also cleaves sialic
acid residues from viral proteins, preventing aggregation of
viruses.
[0074] Ideally, influenza virus neuraminidase (NA) should act on
the same type of receptor the virus hemagglutinin (HA) binds to, a
phenomenon which does not always happen. It is not quite clear how
the virus manages to function when there is no close match between
the specificities of NA and HA.
[0075] The viral neuraminidase enzyme can have endo- or
exo-glycosidase activity, and are classified as EC 3.2.1.29
(endo-neuraminidase) and EC 3.2.1.18 (exo-neuraminidases). In
general, mammalian sialic acid residues are at terminal positions
(non-reducing end) in complex glycans, and so viral neuraminidases
which are exo-glycosidase enzymes use these terminal residues as
their substrates.
[0076] Influenza viruses bind through hemagglutinin onto sialic
acid sugars on the surfaces of epithelial cells; typically in the
nose, throat and lungs of mammals and intestines of birds (Stage 1
in infection). After the hemagglutinin is cleaved by a protease,
the cell imports the virus by endocytosis.
[0077] Once inside the cell, the acidic conditions in the endosome
cause two events to happen: first part of the hemagglutinin protein
fuses the viral envelope with the vacuole's membrane, then the M2
ion channel allows protons to move through the viral envelope and
acidify the core of the virus, which causes the core to dissemble
and release the viral RNA and core proteins. The viral RNA (vRNA)
molecules, accessory proteins and RNA-dependent RNA polymerase are
then released into the cytoplasm (Stage 2). The M2 ion channel is
blocked by amantadine drugs, preventing infection.
[0078] These core proteins and vRNA form a complex that is
transported into the cell nucleus, where the RNA-dependent RNA
polymerase begins transcribing complementary positive-sense vRNA
(Steps 3a and b). The vRNA is either exported into the cytoplasm
and translated (step 4), or remains in the nucleus. Newly
synthesized viral proteins are either secreted through the Golgi
apparatus onto the cell surface (in the case of neuraminidase and
hemagglutinin, step 5b) or transported back into the nucleus to
bind vRNA and form new viral genome particles (step 5a). Other
viral proteins have multiple actions in the host cell, including
degrading cellular mRNA and using the released nucleotides for vRNA
synthesis and also inhibiting translation of host-cell mRNAs.
[0079] Negative-sense vRNAs that form the genomes of future
viruses, RNA-dependent RNA polymerase, and other viral proteins are
assembled into a virion. Hemagglutinin and neuraminidase molecules
cluster into a bulge in the cell membrane. The vRNA and viral core
proteins leave the nucleus and enter this membrane protrusion (step
6). The mature virus buds off from the cell in a sphere of host
phospholipid membrane, acquiring hemagglutinin and neuraminidase
with this membrane coat (step 7). As before, the viruses adhere to
the cell through hemagglutinin; the mature viruses detach once
their neuraminidase has cleaved sialic acid residues from the host
cell. Drugs that inhibit neuraminidase, such as oseltamivir,
therefore prevent the release of new infectious viruses and halt
viral replication. After the release of new influenza viruses, the
host cell dies.
[0080] Because of the absence of RNA proofreading enzymes, the
RNA-dependent RNA polymerase that copies the viral genome makes an
error roughly every 10 thousand nucleotides, which is the
approximate length of the influenza vRNA. Hence, the majority of
newly manufactured influenza viruses are mutants, this causes
"antigenic drift", which is a slow change in the antigens on the
viral surface over time. The separation of the genome into eight
separate segments of vRNA allows mixing or reassortment of vRNAs if
more than one type of influenza virus infects a single cell. The
resulting rapid change in viral genetics produces antigenic shifts,
which are sudden changes from one antigen to another. These sudden
large changes allow the virus to infect new host species and
quickly overcome protective immunity.
[0081] New research shows that the 2009 H1N1 "swine flu" virus may
lack a piece of genetic material that has been present in all other
pandemic flu viruses, meaning that this flu may not be quite as
transmissible as other flu viruses. However, in two instances
discovered thus far, one in Denmark and one in Japan, the virus has
been found to be resistant to antiviral drugs, which is concerning
as these drugs may provide some defense if taken early on in the
illness.
[0082] New mutations in the H1N1 virus are believed to allow it to
flourish in the small intestine, which is something that "normal"
influenza cannot do. Researchers believe this is why the H1N1 virus
can cause gastrointestinal symptoms, such as nausea, vomiting and
diarrhea. Seasonal influenza typically causes respiratory symptoms
without the gastrointestinal symptoms seen in the H1N1 virus.
[0083] Accordingly, while not bound by theory, the etiology of
influenza susceptibility may be caused, in part, by an inadequate
response of the human gastrointestinal immune system, e.g., in
populations such as the elderly and children, and in
immune-compromised individuals.
[0084] Given the above, it is a goal of the present disclosure to
provide therapeutic methods and pharmaceutical compositions for the
prevention and treatment of influenza by supplementing the normal
pancreatic functions and functions of the gastrointestinal immune
system with digestive enzyme preparations, including the
pharmaceutical compositions described herein.
[0085] Another goal of the present disclosure is the use of
pancreatic enzymes to provide therapeutic methods and
pharmaceutical compositions for the prevention and treatment of
Influenza A, Subtype H1N1 swine flu.
[0086] Another goal of the present disclosure is the use of avian
proventriculus and small intestine enzymes to provide therapeutic
methods and pharmaceutical compositions for the prevention of
Influenza A, Subtype H1N1 avian flu.
[0087] Another goal of the present disclosure is the provision of
pharmaceutical compositions for the prevention and/or treatment of
the Influenza, wherein the compositions comprise one or more
digestive enzymes, e.g., one or more enzymes selected from
amylases, proteases, cellulases, papain (e.g., from papapya),
bromelain (e.g., from pineapples), lipases, chymotrypsin, trypsin,
and hydrolases. In some embodiments, the pharmaceutical
compositions are lipid encapsulated.
[0088] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for pregnant women,
neonates, and infants.
[0089] Another goal of the present disclosure is to provide a
treatment for the symptoms of influenza for pregnant women,
neonates, and infants.
[0090] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for the elderly, including
those that do not respond well to vaccination.
[0091] Another goal of the present disclosure is to provide a
treatment for the symptoms of influenza in the elderly.
[0092] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for HTV compromised
individuals and those with ATDS.
[0093] Another goal of the present disclosure is to provide a
treatment for the symptoms of influenza in HIV compromised
individuals and those with AIDS.
[0094] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for organ transplant
recipients.
[0095] Another goal of the present disclosure is to provide a
treatment for the symptoms of influenza for organ transplant
recipients.
[0096] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for persons with any
underlying medical conditions including asthma, reactive airways
disease, or other chronic disorders of the pulmonary or
cardiovascular systems; other underlying medical conditions,
including such metabolic diseases as diabetes, renal dysfunction,
and hemoglobinopathies; or known or suspected immunodeficiency
diseases or immunosuppressed states.
[0097] Another goal of the present disclosure is to provide a
treatment for the symptoms of influenza for persons with underlying
medical conditions including asthma, reactive airways disease, or
other chronic disorders of the pulmonary or cardiovascular systems;
other underlying medical conditions, including such metabolic
diseases as diabetes, renal dysfunction, and hemoglobinopathies; or
known or suspected immunodeficiency diseases or immunosuppressed
states.
[0098] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for children or adolescents
receiving aspirin or other salicylates due to increased risk of
Reye Syndrome.
[0099] Another goal of the present disclosure is to provide a
treatment for the symptoms of influenza for children or adolescents
receiving aspirin or other salicylates due to increased risk of
Reye Syndrome.
[0100] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for individuals with acute
febrile illness.
[0101] Another goal of the present disclosure is to provide a
treatment for the symptoms of influenza for individuals with acute
febrile illness.
[0102] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for children with chronic
medical conditions and children under the age of 2.
[0103] Another goal of the present disclosure is to provide a
treatment for the symptoms of influenza for children with chronic
medical conditions and children under the age of 2.
[0104] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for individuals with
hypersensitivity and allergic reactions including hypersensitivity
and allergies to residual egg proteins and other components of TIV
and LAIV influenza vaccines including thimerosal and
antibiotics.
[0105] Another goal of the present disclosure is to provide a
prophylactic measure against influenza for those with a
proportionately higher risk of Guillain-Barre Syndrome recurrence
with TIV or LAW influenza vaccination or those with the inability
to use LAIV after a history of GBS with TIV.
[0106] Another goal of the present disclosure is to provide a
prophylactic measure against influenza with reduced risk of
Guillain-Barre Syndrome.
[0107] Another goal of the present disclosure is to provide a
prophylactic measure against influenza which reduces or eliminates
viral shedding.
[0108] Another goal of the present disclosure is to provide a
treatment for influenza which reduces or eliminates viral
shedding.
[0109] Another goal of the present disclosure is to provide a
prophylactic measure against influenza which reduces or eliminates
LAIV and TIV side effects including, but not limited to: runny
nose, headache, vomiting, myalgias, abdominal pain sore throat,
asthma, and tiredness/weakness.
[0110] Another goal of the present disclosure is to provide a
prophylactic measure against influenza which does not require
forward prediction of virus strains and associated antigens to be
effective.
[0111] Another goal of the present disclosure is to provide a
prophylactic measure against influenza which does not require
extensive prediction of virus strains and associated antigens to
have extensive lead time to produce vaccine in sufficient
quantities.
[0112] Another goal of the present disclosure is to provide a
prophylactic measure against influenza with inherently lower risk
factors thereby increasing prophylactic coverage of both at risk
and general populations, thereby overcoming the reluctance of
individuals who refuse to obtain a LAIV or TIV vaccination.
[0113] Another goal of the present disclosure is to provide a
prophylactic measure against influenza which does not promote or
contribute to drug resistance.
[0114] An additional goal of the disclosure is to demonstrate the
use of fecal chymotrypsin level as a biomarker for the likelihood
of an individual's susceptibility to influenza.
[0115] Yet another goal of the disclosure is to demonstrate the use
of fecal chymotrypsin level as a biomarker for diagnosis of a
compromised immune system, or for determining if an individual is
likely to benefit from administration of a composition as described
herein.
[0116] Yet another goal is to provide adjunctive compositions and
methods to vaccination and/or anti-viral prophylactic medications,
and/or adjunctive compositions and methods to therapeutic
medications for the prevention and treatment of influenza.
[0117] Tt is a further goal of the present disclosure to provide
viricidal and/or viristatic compositions comprising one or more
digestive enzymes for use as or in disinfectants, sanitizers,
detergents, and antiseptics, e.g., in hospitals, nursing homes,
nurseries, daycares, schools, work environments, public
transportation and restroom facilities, to reduce and/or destroy
influenza viruses present in such settings. The surfaces can be
large (e.g., operating room tables, doors, changing tables) or
small (e.g., medical devices, door handles); inanimate (tables) or
animate (hands, e.g., detergents for hand-washing). The
compositions can thus be useful to treat surfaces to reduce or kill
influenza virus thereon, and thereby prevent or reduce the spread
of influenza.
[0118] Accordingly, provided herein is a method for preventing
and/or treating Influenza comprising administering to the patient a
therapeutically effective amount of a pharmaceutical composition
comprising one or more digestive enzymes. In some embodiments, the
pharmaceutical composition may be encapsulated; such encapsulated
compositions may be referred to as enzyme preparations herein. In
some embodiments, the pharmaceutical composition can be
lipid-encapsulated.
[0119] In some embodiments, the one or more digestive enzymes
comprise one or more enzymes selected from the group consisting of
proteases, amylases, celluloses, sucrases, maltases, papain (e.g.,
from papaya), bromelain (e.g., from pineapple), hydrolases, and
lipases. In some embodiments, the one or more digestive enzymes
comprise one or more pancreatic enzymes. In some embodiments, the
pharmaceutical composition comprises one or more proteases, one or
more lipases, and one or more amylases. In some embodiments, the
one or more proteases comprise chymotrypsin and trypsin.
[0120] The one or more digestive enzymes are, independently,
derived from an animal source, a microbial source, a fungal source,
or a plant source, or are synthetically prepared. In some
embodiments, the animal source is a pig, e.g., a pig pancreas, or
avian, e.g., "bird" proventriculus or small intestine.
[0121] In some embodiments, the pharmaceutical composition
comprises at least one amylase, a mixture of proteases comprising
chymotrypsin and trypsin, and at least one lipase. The
pharmaceutical composition can further include papain, e.g., from
papaya. In some embodiments, the pharmaceutical composition
comprises per dose: amylases from about 10,000 to about 60,000
U.S.P, including 10,000, 15,000, 20,000, 25,000, 30,000, 35,000,
40,000, 45,000, 50,000, 55,000, and 60,000 U.S.P, along with all
values in-between, proteases from about 10,000 to about 70,000
U.S.P, including 10,000, 15,000, 20,000, 25,000, 30,000, 35,000,
40,000, 45,000, 50,000, 55,000, 60,000, 65,000, and 70,000, along
with all values in-between, lipases from about 4,000 to about
30,000 U.S.P, including, 4,000, 5,000, 10,000, 15,000, 20,000,
25,000, and 30,000, along with all values in-between, chymotrypsin
from about 2 to about 5 mg including 2.0, 2.5, 3.0, 3.5, 4.0, 4.5,
and 5.0 mg, along with all values in-between, trypsin from about 60
to about 100 mg including 50, 65, 70, 75, 80, 85,90, 95, and 100
mg, including all values in between; papain from about 3,000 to
about 10,000 USP units including 3,000, 4,000, 5,000, 6,000, 7,000,
8,000, 9,000, and 10,000 USP, along with all values in between, and
papaya from about 30 to about 60 mg, including 30, 35, 40, 45, 50,
55, and 60 mg, along with all values in between.
[0122] In some embodiments, the pharmaceutical composition
comprises at least one protease and at least one lipase, wherein
the ratio of total proteases to total lipases (in USP units) ranges
from about 1:1 to about 20:1 including 1:1, 2:1, 3;1, 4:1, 5;1,
6:1, 7:1, 8:1, 9:1, 10:1, 11;1, 12;1, 13;1, 14:1, 15:1, 16;1, 17:1,
18:1, 19:1 and 20:1, long with all values in-between. In some
embodiments, the ratio of proteases to lipases ranges from about
4:1 to about 10:1 including 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, and 10:1,
along with all values in-between.
[0123] In some embodiments, the one or more symptoms of influenza
include: fever, headache, tiredness, cough, sore throat, runny or
stuffy nose, body aches, diarrhea and vomiting, and a combination
thereof.
[0124] In some embodiments, the pharmaceutical composition is a
dosage formulation selected from the group consisting of: pills,
tablets, capsules, microcapsules, mini-capsules, time released
capsules, mini-tabs, sprinkles, and a combination thereof.
[0125] In some embodiments, a pharmaceutical composition comprises
a core comprising one or more digestive enzymes, as described
previously and a coating which comprises a crystallizable lipid.
The core contains an amount of the one or more digestive enzymes
effective for the prevention or treatment of Influenza. Among other
properties, the coating protects the digestive enzymes from
destabilizing factors such as solvents, heat, light, moisture and
other environmental factors. The coating also provides controlled
release of the enzymes when the encapsulate is exposed to a
physiological conditions. In addition, in one aspect of this
disclosure, the coated digestive enzyme preparations of this
disclosure have improved pour properties, and improved taste and
smell of the digestive enzyme particles. The coated digestive
enzyme preparations can be used to obtain release at selected
transit times or in selected locations of the gastrointestinal
tract of mammals, as necessary.
[0126] In some embodiments, a specific blend of enzymes and lipids
for enzyme administration to individuals for the prevention and/or
treatment or influenza is provided.
[0127] In another embodiment 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, including 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,
45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, and 90% by weight,
along with all values in-between; and (b) a coating comprising a
crystallizable lipid, wherein the coating continuously coats the
core and the crystallizable lipid releases the enzyme upon exposure
to physiological conditions.
[0128] In another embodiment 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 prophylaxis
of influenza or treatment of the symptoms of influenza; and (b)
coating comprising a crystallizable lipid.
[0129] In yet another embodiment an enzyme delivery system
comprising an encapsulated enzyme preparation having particles
which comprise: (a) a core comprising pancreatic or digestive
enzymes present in an amount of from about 5% to 95% by weight of
the particles, including 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,
45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% and 95% by weight
along with all values in-between; and (b) a generally uniform
coating to provide for controlled release of the enzymes, said
coating comprising a crystallizable lipid. In yet another
embodiment, the encapsulated enzyme preparation particles of the
enzyme delivery system are non-aerosolizable.
[0130] In certain embodiments, the enzyme preparations according to
this disclosure 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.
[0131] Certain coated digestive enzyme preparations which comprise
a coating of a crystallizable 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 prevention or treatment of influenza with digestive
enzymes. In some aspects, the coated pancreatic/digestive enzyme
preparations are prepared to obtain specific delivery times or
specific regions within the human gastrointestinal (GI) tract. In
some embodiments, the crystallizable lipid composition is
hydrogenated soybean oil, but may be any suitable crystallizable
lipid or lipid blend.
[0132] Some embodiments utilize 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 prophylaxis or treatment.
[0133] In another embodiment enzyme preparations are utilized with
lipid coating and/or encapsulation of enzymes. The method of making
the preparations comprises providing a crystallizable lipid, and
coating screened pancreatic/digestive enzyme particles as described
herein with the lipid. The digestive enzymes comprise 5% to 95% by
weight of the coated enzyme preparations, including 5%, 10%, 15%,
20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,
85%, 90% and 95%, along with all values in-between;
[0134] The methods of this disclosure can be used to produce coated
digestive enzyme preparations comprising digestive and/or
pancreatic enzymes coated with a crystallizable 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.
Encapsulated pancreatic/digestive enzyme preparations having a
coating comprising 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 HCl.
[0135] In another embodiment, a method for administering the enzyme
preparations includes administering compositions comprising one or
more digestive enzymes as coated preparations. In some aspects, the
disclosure relates to a prophylactic method comprising
administering to a subject 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 a crystallizable 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, and/or
exhibits one or more symptoms associated with Influenza, and/or is
diagnosed with Influenza, and/or is immune-compromised.
[0136] In another embodiment the pancreatic/digestive enzyme
composites, preparations, enzyme delivery compositions or systems
comprise no or fewer excipients, carriers, additives and/or
extenders, and/or require the use of no or fewer solvents. In some
embodiments, the coating comprises, consists of, or consists
essentially of hydrogenated soybean oil, reducing exposure to
potentially toxic substances and also reducing the possibility of
allergy formation. In some embodiments the delivery of pancreative
and/or digestive enzymes has improved safety of administration.
[0137] Also provided are methods for diagnosing patients based on
fecal chymotrypsin levels. In one embodiment a method of diagnosing
a patient comprises: obtaining a fecal sample from the patient,
determining a level of chymotrypsin present in the fecal sample, in
some cases wherein the determination is performed at 30.degree. C.,
and diagnosing the patient as having a compromised immune system if
the determined chymotrypsin level is less than a normal level
(e.g., a control level).
[0138] In some embodiments, the fecal chymotrypsin level is between
8.4 and 4.2 U/gram. In some embodiments, the fecal chymotrypsin
level is less than 4.2 U/gram. In some embodiments, the level of
chymotrypsin present in the fecal sample is determined using an
enzymatic photospectrometry method. In some embodiments, the method
further comprises administering to the patient an effective amount
of a pharmaceutical composition comprising one or more digestive
enzymes if the patient is determined to have a compromised immune
system, e.g., to treat or prevent influenza in the individual.
[0139] In some embodiments, the method further comprises
determining if the administration of the pharmaceutical composition
reduces the effects or symptoms of Influenza.
[0140] Also provided is a method of identifying a patient likely to
benefit from administration of a pharmaceutical composition
comprising one or more digestive enzymes comprising: obtaining a
fecal sample from the patient, determining a level of chymotrypsin
present in the fecal sample, in some cases wherein the
determination is performed at 30.degree. C., and identifying the
patient as likely to benefit from administration of the
pharmaceutical composition if the determined fecal chymotrypsin
level is less than a normal (e.g., control level), e.g., in some
embodiments 8.4 U/gram or less. In some embodiments, the patient is
diagnosed with Influenza and/or is immune compromised. In some
embodiments, the method further comprises determining if the
patient exhibits one or more symptoms of Influenza. In some
embodiments, the benefit comprises a reduction or amelioration of
one or more symptoms associated with the Influenza. In some
embodiments, the method further comprises administering to the
patient an effective amount of a pharmaceutical composition
comprising one or more digestive enzymes.
[0141] Also provided is a pharmaceutical composition comprising one
or more digestive enzymes, wherein the one or more digestive
enzymes comprise at least one lipase and at least one protease, and
wherein the ratio of total proteases to total lipases (in USP
units) ranges from about 1:1 to about 20:1 including 1:1, 2:1, 3;1,
4:1, 5;1, 6:1, 7:1, 8:1, 9:1, 10:1, 11;1, 12;1, 13;1, 14:1, 15:1,
16;1, 17:1, 18:1, 19:1 and 20:1 long with all values in-between. In
some embodiments, the ratio of total proteases to total lipases
ranges from about 4:1 to about 10:1 including 4:1, 5:1, 6:1, 7:1,
8:1, 9:1, and 10:1 along with all values in-between. In some
embodiments, the pharmaceutical composition is lipid
encapsulated.
[0142] Also provided is a pharmaceutical composition comprising at
least one amylase, a mixture of proteases comprising chymotrypsin
and trypsin, at least one lipase, and optionally papaya and/or
papain. In some embodiments, the ratio of total proteases to total
lipases ranges from about 1:1 to about 20:1 including 1:1, 2:1,
3;1, 4:1, 5;1, 6:1, 7:1, 8:1, 9:1, 10:1, 11;1, 12;1, 13;1, 14:1,
15:1, 16;1, 17:1, 18:1, 19:1 and 20:1 along with all values
in-between.
[0143] The features and advantages described herein are not
all-inclusive and, in particular, many additional features and
advantages will be apparent to one of ordinary skill in the art in
view of the drawings, specification, and claims.
DETAILED DESCRIPTION
[0144] The present disclosure provides pharmaceutical compositions
comprising one or more digestive enzymes and methods of using the
same for the treatment and/or prevention of Influenza. The present
disclosure also provides compositions comprising one or more
digestive enzymes and methods of using the same as antiseptics,
detergents, disinfectants, and sanitizers, e.g., as viricidal
and/or viristatic compositions to kill or attenuate the influenza
virus. The compositions described herein include one or more
digestive enzymes, which are postulated to assist in the reduction,
weakening, or eradication of Influenza virus, and thus to prevent
contraction of Influenza; and/or to ameliorate gastrointestinal
dysfunction or to enhance the normal gastrointestinal function, in
order to prevent contraction of Influenza or to treat Influenza
(e.g., improve or ameliorate the symptoms or reduce the time course
of the infection). In addition, the pharmaceutical compositions can
be utilized to enhance immune system response for individuals with
compromised immune systems and/or to augment immune system
functions is nonimmuno-compromised individuals, e.g., to assist in
the prevention or treatment of Influenza.
[0145] In certain embodiments, the pharmaceutical compositions can
include one or more digestive enzymes, wherein the one or more
digestive enzymes comprise at least one lipase and at least one
protease, and wherein the ratio of total proteases to total lipases
(in USP units) ranges from about 1:1 to about 20:1, including 1:1,
2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1,
14:1, 15:1, 16:1, 17:1, 18:1, 19:1 and 20:1 along with all values
in-between. In some cases, the ratio of total proteases to total
lipases ranges from about 4:1 to about 10:1, including 4;1, 4;1,
6:1, 7:1, 8:1, 9:1, and 10:1 along with all values in-between. In
some embodiments, the pharmaceutical composition is encapsulated,
e.g., lipid-encapsulated. Enzyme preparations comprising one or
more digestive enzymes useful for the methods described herein are
disclosed in U.S. Ser. No. 12/386,051, incorporated herein by
reference.
[0146] In some cases, a pharmaceutical composition for use herein
comprises at least one amylase, at least one protease, and at least
one lipase. In certain embodiments, the composition can comprise at
least one amylase, at least two proteases, and at least one lipase.
In certain embodiments the pharmaceutical composition includes
multiple proteases, including, without limitation, chymotrypsin and
trypsin. In certain embodiments, the composition can further
include one or more hydrolases, papain, bromelain, papaya,
celluloses, pancreatin, sucrases, and maltases.
[0147] The one or more enzymes can be independently derived from
animal, plant, fungal, microbial, or synthetic sources. In some
embodiments, the one or more enzymes are derived from pig, e.g. pig
pancreas or avian "bird" proventriculus or small intestine.
[0148] One exemplary formulation for the prevention of Influenza or
treatment of symptoms of Influenza is as follows:
Amylase 10,000-60,000 U.S.P
Protease 10,000-70,000 U.S.P
Lipase 4,000-30,000 U.S.P
Chymotrypsin 2-5 mg
Trypsin 60-100 mg
[0149] Papain 3,000-10,000 USP units/mg
Papaya 30-60 mg
[0150] Additional formulations comprising one or more digestive
enzymes may be advantageous including formulations in which the
ratio of total proteases to total lipases (in USP units) is from
about 1:1 to about 20:1, including 1:1, 2:1, 3:1, 4:1, 5:1, 6:1,
7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1,
18:1, 19:1 and 20:1 along with all values in-between. In some
embodiments, the ratio of total proteases to total lipases is from
about 4:1 to about 10:1 including 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, and
10:1 along with all values in-between. Such formulations are useful
for the prevention of Influenza or treating symptoms of Influenza,
or the enhancement of immune system functions.
[0151] The pharmaceutical compositions can be formulated in dosage
forms for any route of administration, including oral, parenteral,
IV, inhalation, and buccal dosage formulations. In certain
embodiments, a dosage formulation may be administered by an oral
preparation including, but not limited to, an encapsulated tablet,
mini-tabs, microcapsule, mini-capsule, time released capsule,
sprinkle, powder or other methodology. In one embodiment, the oral
preparation is encapsulated using one or more lipids.
Alternatively, the oral preparation may be encapsulated using
enteric coating or organic polymers. A formulation may also be
prepared using Prosolv.RTM. technology, direct compression, dry
granulation, wet granulation, and/or a combination of these
methods.
[0152] Fecal chymotrypsin level is a sensitive, specific measure of
proteolytic activity, see, e.g. U.S. Pat. No. 6,660,831,
incorporated by reference herein. Normal levels of chymotrypsin are
typically considered to be greater than 8.4 U/gram. Decreased
values (less than 4.2 U/gram) suggest diminished pancreatic output
(pancreatic insufficiency), hypoacidity of the stomach or cystic
fibrosis. Elevated chymotrypsin values suggest rapid transit time,
or less likely, a large output of chymotrypsin from the
pancreas.
[0153] For the fecal chymotrypsin test, a stool sample can be
collected from each of the subjects. Each stool sample can be
analyzed using an enzymatic photospectrometry analysis to determine
the level of fecal chymotrypsin in the stool; in some cases the
assay is performed at 30.degree. C., see, e.g. U.S. Pat. No.
6,660,831, incorporated by reference herein. Alternatively, other
methods, such as the colorimetric method, use of substrates, use of
assays, and/or any other suitable method may be used to measure the
fecal chymotrypsin levels. The levels of fecal chymotrypsin in the
samples e.g., of individuals suspected of or diagnosed as having a
compromised immune system, are compared to the levels of fecal
chymotrypsin in normal or control individuals (e.g., individuals
not suspected or diagnosed with a compromised immunes system), to
determine if the individuals would benefit from the administration
of a composition as described herein.
[0154] The nature of the human digestive tract creates challenges
for the delivery of digestive enzymes to patients including the
general population, those with compromised immune systems, or those
with influenza. 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, calls for a specific delivery method
depending upon where the enzyme is to be delivered. For veterinary
applications, a specific delivery location may also be necessary,
depending on the animal to be treated.
[0155] 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 destabilization 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 compositions from unfavorable conditions, the
digestive enzymes may be coated or encapsulated in a continuous
coating containing a crystallizable lipid.
[0156] Manufacturers of enzyme preparations have used enteric
coatings to deliver lipases in individuals requiring administration
of lipases, such as individuals with cystic fibrosis.
[0157] 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.
[0158] For example, a lipid coating of this disclosure 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
disclosure, 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.
[0159] 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 uniformly
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.
[0160] As discussed, the encapsulation can provide controlled
release of the digestive enzymes. 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. In some embodiments,
the dissolution profile may 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.
[0161] "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.
[0162] The crystallizable lipid is any lipid or wax, lipid or wax
mixture, or blend of lipid and/or waxes, where the crystalliable
lipid forms a solid coating via crystallization at typical storage
temperatures. The crystallizable lipid can be a vegetable or animal
derived-lipid. In some embodiments, the crystallizable lipid is
emulsifiable upon contact with physiological conditions and
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 crystallizable lipid is a non-polar lipid,
for example hydrogenated soybean oil
[0163] 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.
[0164] The digestive enzyme used in the present disclosure can be,
for example, 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
disclosure is not limited to pancreatic enzymes of porcine origin,
but can be of other animal, microbial, 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, fungal or mammalian cells, and can include a
mixture of enzymes from one or more sources. Digestive enzymes, 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.
[0165] In some embodiments, the digestive enzyme particles used as
cores in the present disclosure 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, including
105, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400,
and 425 um 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 disclosure 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.
[0166] The minimum amount of pancreatic enzyme present in the core
can vary, and in some embodiments 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 can vary, and in some cases 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.
[0167] 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.
[0168] Further, in some embodiments, the lipid encapsulation
methodology reduces the aerosolization of the enzyme preparation
that may be caustic to an individual if inhaled through the lungs
or the nose. The lipid encapsulation reduces aerolization and the
potential for caustic burn, aspiration, and/or aspiration
pneumonias in receivers and administrators of the enzyme
preparation, thereby reducing the potential for illness in those
already compromised by influenza or reduced immune system
functionality, and leading to safer administration.
[0169] 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).
[0170] 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 individual
to be treated.
[0171] Additives can be blended with a crystallizable 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 can be chosen to release the
bioactive substance in the area of the digestive tract selected for
release to optimize treatment.
[0172] The disclosure 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 disclosure 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.
[0173] 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.
[0174] In other embodiments, the disclosure relates to methods of
treatment comprising administering to a subject, at least two doses
of a composition comprising a therapeutically effective amount of
the coated digestive enzyme preparations for the prophylaxis or
treatment or influenza. 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.
[0175] In other embodiments, the disclosure relates to methods of
treatment comprising administering to a subject, at least three
doses of a composition comprising a therapeutically effective
amount of the coated digestive enzyme preparations for the
prophylaxis or treatment or influenza. 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 reaches the small intestine.
[0176] Another embodiment of the disclosure 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 soybean oil.
[0177] The lipid coating surprisingly does not appear to be reduced
or destroyed by HCl (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, an excipient containing only lipid can be used to coat or
encapsulate digestive enzyme particles containing lipase.
[0178] The disclosure therefore relates to improvement of the
delivery of digestive enzymes to humans or animals based
specifically upon needed delivery times, and dissolution profiles.
For example, in certain aspects of the disclosure, the rate of
release and dissolution characteristics are unique to the lipid
encapsulations of this disclosure
[0179] For prophylaxis of influenza or treatment of patients with
influenza or immune compromised individuals 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 virion protein
digestion. In another example, for elderly patients with slower GI
transit times, still another release profile may be advantageous to
deliver enzymes for effective treatment. The lipid and/or additive
selection will be made to obtain enzyme release at later times
after administration. In veterinary applications, still another
release profile may be necessary.
[0180] The present disclosure 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
a crystallizable lipid, and coating pancreatic/digestive enzyme
particles with the lipid, where the pancreatic/digestive enzymes
comprise 5-90% including 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55,
60, 65, 70, 75, 80, 85, and 90% along with all values in-between 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 including 105, 125, 150, 175, 200, 225, 250,
275, 300, 325, 350, 375, 400, and 425 um along with all values
in-between.
[0181] In one embodiment, the disclosure 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 a
crystallizable lipid to form coated or encapsulated digestive
enzymes containing a core which contains the pancreatic/digestive
enzyme and a coating which contains the crystallizable lipid. In
some embodiments, the encapsulated digestive enzyme preparation is
a controlled release digestive enzyme preparation, which may have
enhanced flow properties.
[0182] 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.
[0183] 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 crystallizable lipid can be any lipid or lipid-derived material
that creates a solid coating around the digestive enzyme substrate.
This lipid coating releases the digestive enzyme in the GI tract.
The lipid coating may be emulsifiable in the GI tract, releasing
the digestive enzyme.
[0184] The crystallizable 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 and waxes. In some embodiments,
the lipid is hydrogenated. The lipid can also be saturated or
partially saturated. Examples of crystallizable lipids include, but
are not limited to, monoglycerides, diglycerides, triglycerides,
fatty acids, esters of fatty acids, phospholipids, salts thereof,
and combinations thereof. The crystallizable lipid can be an
emulsifiable lipid.
[0185] "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.
[0186] The crystallizable lipid which is emulsifiable is preferably
a food grade emulsifiable lipid. Some examples of food grade
emulsifiable lipids include sorbitan monostearates, sorbitan
tristcaratcs, 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.
[0187] Any emulsifiable lipid may be used in the methods and
products of this disclosure. In certain embodiments the
emulsifiable lipid used will produce non-agglomerating,
non-aerosolizing enzyme preparation particles.
[0188] The coating of the enzyme with the lipid 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 appropriate containers.
[0189] The inclusion of one or more additives with an emulsifiable
lipid of the present disclosure 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.
[0190] 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.
[0191] 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.
[0192] 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.
[0193] 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.
[0194] Other additives for inclusion in the compositions described
herein can be determined by those having ordinary skill in the art,
and will be based on a number of features, including intended
application, e.g., human vs. veterinary applications; desired
release profile; desired pharmacokinetics; safety; stability;
physical characteristics (smell, color, taste, pour,
aerosilization). Suitable formulation ingredients, excipients,
binders, bulking agents, flavorants, colorants, etc. can be
determined and evaluated by methods known to those having ordinary
skill.
[0195] In one aspect of the disclosure, the method comprises using
the enzyme formulations to treat individuals that 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.
[0196] The present disclosure also provides viricidal and/or
viristatic compositions comprising one or more digestive enzymes
for use as or in disinfectants, sanitizers, detergents, and
antiseptics, e.g., in hospitals, nursing homes, nurseries,
daycares, schools, work environments, public transportation and
restroom facilities, to reduce and/or destroy influenza viruses
present in such settings. The surfaces can be large (e.g.,
operating room tables, doors, changing tables) or small (e.g.,
medical devices, door handles); inanimate (tables) or animate
(hands, e.g., detergents for hand-washing). The compositions
include one ore more digestive enzymes, and optionally additives
customarily used in antiseptics, disinfectants, sanitizers and
detergents. The viricidal and viristatic compositions can be
encapsulated enzyme compositions, as described previously, to
increase stability and shelf-life.
[0197] Disinfectants are antimicrobial agents that are applied to
non-living objects to destroy microorganisms, the process of which
is known as disinfection. Disinfectants should generally be
distinguished from antibiotics that destroy microorganisms within
the body, and from antiseptics, which destroy microorganisms on
living tissue. Sanitizers are substances that reduce the number of
microorganisms to a safe level. One official and legal version
states that a sanitizer must be capable of killing 99.999%, known
as a 5 log reduction, of a specific test population, and to do so
within 30 seconds. The main difference between a sanitizer and a
disinfectant is that at a specified use dilution, the disinfectant
must have a higher kill capability compared to that of a sanitizer.
Very few disinfectants and sanitizers can sterilize (the complete
elimination of all microorganisms), and those that can depend
entirely on their mode of application.
[0198] The choice of the disinfectant to be used depends on the
particular situation. Some disinfectants have a wide spectrum (kill
nearly all microorganisms), whilst others kill a smaller range of
disease-causing organisms but are preferred for other properties
(they may be non-corrosive, non-toxic, or inexpensive).
[0199] The relative effectiveness of disinfectants can be measured
by comparing how well they do against a known disinfectant and rate
them accordingly. Phenol is the standard, and the corresponding
rating system is called the "Phenol coefficient". The disinfectant
to be tested is compared with phenol on a standard microbe (usually
Salmonella typhi or Staphylococcus aureus). Disinfectants that are
more effective than phenol have a coefficient >1. Those that are
less effective have a coefficient <1. For example, the
Rideal-Walker method gives a Rideal-Walker coefficient and the U.S.
Department of Agriculture method gives a U.S. Department of
Agriculture coefficient.
[0200] To calculate phenol coefficient, the concentration of the
test compound at which the compound kills the test organism in 10
minutes, but not in 5 minutes, is divided by the concentration of
phenol that kills the organism under the same conditions. The
phenol coefficient may be determined in the presence of a standard
amount of added organic matter or in the absence of organic
matter.
[0201] A detergent is a material intended to assist cleaning. The
term is sometimes used to differentiate between soap and other
surfactants used for cleaning where soap is a surfactant cleaning
compound, typically used for personal or minor cleaning.
[0202] Detergents and soaps are used for cleaning because pure
water can't remove oily, organic soiling. Soap cleans by acting as
an emulsifier. Basically, soap allows oil and water to mix so that
oily grime can be removed during rinsing. Detergents were developed
in response to the shortage of the animal and vegetable fats used
to make soap during World War I and World War II. Detergents are
primarily surfactants, which could be produced easily from
petrochemicals. Surfactants lower the surface tension of water,
essentially making it `wetter` so that it is less likely to stick
to itself and more likely to interact with oil and grease.
[0203] Modern detergents contain more than surfactants. Cleaning
products may also contain enzymes to degrade protein-based stains,
bleaches to de-color stains and add power to cleaning agents, and
blue dyes to counter yellowing. Like soaps, detergents have
hydrophobic or water-hating molecular chains and hydrophilic or
water-loving components. The hydrophobic hydrocarbons are repelled
by water, but are attracted to oil and grease. The hydrophilic end
of the same molecule means that one end of the molecule will be
attracted to water, while the other side is binding to oil. Neither
detergents nor soap accomplish anything except binding to the soil
until some mechanical energy or agitation is added into the
equation. Swishing the soapy water around allows the soap or
detergent to pull the grime away from clothes or dishes and into
the larger pool of rinse water. Rinsing washes the detergent and
soil away. Warm or hot water melts fats and oils so that it is
easier for the soap or detergent to dissolve the soil and pull it
away into the rinse water. Detergents are similar to soap, but they
are less likely to form films (soap scum) and are not as affected
by the presence of minerals in water (hard water).
[0204] Detergents, especially those made for use with water, often
include different components such as Surfactants to `cut`
(dissolve) grease and to wet surfaces, abrasives to scour,
substances to modify pH or to affect performance or stability of
other ingredients, acids for descaling or caustics to break down
organic compounds, water softeners to counteract the effect of
"hardness" ions on other ingredients, oxidants (oxidizers) for
bleaching, disinfection, and breaking down organic compounds,
non-surfactant materials that keep dirt in suspension, enzymes to
digest proteins, fats, or carbohydrates in stains or to modify
fabric feel, ingredients that modify the foaming properties of the
cleaning surfactants, to either stabilize or counteract foam,
ingredients to increase or decrease the viscosity of the solution,
or to keep other ingredients in solution, in a detergent supplied
as a water solution or gel, ingredients that affect aesthetic
properties of the item to be cleaned, or of the detergent itself
before or during use, such as optical brighteners, fabric
softeners, colors, perfumes, etc., ingredients such as corrosion
inhibitors to counteract damage to equipment with which the
detergent is used, ingredients to reduce harm or produce benefits
to skin, when the detergent is used by bare hand on inanimate
objects or used to clean skin, and preservatives to prevent
spoilage of other ingredients.
[0205] There are several factors that dictate what compositions of
detergent should be used, including the material to be cleaned, the
apparatus to be used, and tolerance for and type of dirt.
[0206] Modern detergents may be made from petrochemicals or from
oleochemicals derived from plants and animals. Alkalis and
oxidizing agents are also chemicals found in detergents. The most
used disinfectants are those applying: active chlorine (i.e.,
hypochlorites, chloramines, dichloroisocyanurate and
trichloroisocyanurate, wet chlorine, chlorine dioxide etc.): active
oxygen (peroxides, such as peracetic acid, potassium persulfate,
sodium perborate, sodium percarbonate and urea perhydrate); iodine
(iodpovidone (povidone-iodine, Betadine), Lugol's solution, iodine
tincture, iodinated nonionic surfactants); concentrated alcohols
(mainly ethanol, 1-propanol, called also n-propanol and 2-propanol,
called isopropanol and mixtures thereof; further, 2-phenoxyethanol
and 1- and 2-phenoxypropanols are used); phenolic substances (such
as phenol (also called "carbolic acid"), cresols (called "Lysole"
in combination with liquid potassium soaps), halogenated
(chlorinated, brominated) phenols, such as hexachlorophene,
triclosan, trichlorophenol, tribromophenol, pentachlorophenol,
Dibromol and salts thereof); cationic surfactants, such as some
quaternary ammonium cations (such as benzalkonium chloride, cetyl
trimethylammonium bromide or chloride, didecyldimethylammonium
chloride, cetylpyridinium chloride, benzethonium chloride) and
others, non-quarternary compounds, such as chlorhexidine,
glucoprotamine, octenidine dihydrochloride etc.); strong oxidizers,
such as ozone and permanganate solutions; heavy metals and their
salts, such as colloidal silver, silver nitrate, mercury chloride,
phenylmercury salts, copper sulfate, copper oxide-chloride etc.
Heavy metals and their salts are the most toxic, and
environment-hazardous bactericides and therefore, their use is
strongly oppressed or canceled; further, also properly concentrated
strong acids (phosphoric, nitric, sulfuric, amidosulfuric,
toluenesulfonic acids) and alkalis (sodium, potassium, calcium
hydroxides), such as of pH<1 or >13, particularly under
elevated temperature (above 60.degree. C.), kills bacteria.
[0207] Antiseptics (from Greek .alpha..nu..tau.{acute over
()}--anti,
`"against"+.sigma..eta..pi..tau..kappa.o.zeta.--septikos,
"putrefactive") are antimicrobial substances that are applied to
living tissue/skin to reduce the possibility of infection, sepsis,
or putrefaction. They should generally be distinguished from
antibiotics that destroy bacteria within the body, and from
disinfectants, which destroy microorganisms found on non-living
objects.
[0208] A composition for use as a sanitizer, detergent,
disinfectant, or antiseptic can, in some cases, be diluted in a
suitable diluent such as saline, phosphate buffered solutions, and
other pH stabilized aqueous solutions, and can be used in
combination with other sanitizers, detergents, disinfectants, or
antiseptics, such as alcohols, quaternary ammonium compounds, boric
acid, chlorhexidine gluconate, iodine, octenidine dihydrochloride,
and sodium chloride.
[0209] Recent USDA Studies in Swine
[0210] Recent experimental test results from the Unites States
Department of Agriculture serve to verify the efficacy of porcine
intestinal immune systems against various H1N1 and other influenza
diseases.
[0211] Experiment 1:
[0212] Four Pig Pathogenesis Study with the 2009 A/H1N1 Influenza A
Virus.
[0213] Purpose of Study:
[0214] An important concern is to address whether meat, blood and
tissue from pigs infected with the new 2009 H1N1 Influenza A Virus
is free of infectious virus.
[0215] Experiment:
[0216] Four 5-week-old cross-bred pigs from a herd free of swine
influenza virus (SIV) and porcine reproductive and respiratory
syndrome virus (PRRSV) were housed in containment facilities and
cared for in compliance with the Institutional Animal Care and Use
Committee of the National Animal Disease Center (NADC).
[0217] Pigs were inoculated intra-tracheally with an infective dose
of the 2009 HIN1 Influenza A Virus isolated from persons in
California (A/CA/04/2009) obtained from the Center for Disease
Control and Prevention (CDC).
[0218] Pigs were observed daily for clinical signs of disease.
Nasal swabs were taken on 0, 1, 2, 3, 4, and 5 days post infection
(dpi) to evaluate nasal shedding. Pigs were humanely euthanized on
5 dpi, which is considered the peak of infection in the NADC
porcine SIV challenge model, to evaluate lung lesions and viral
load in the lung and tissues. Fresh samples were taken from lung,
tonsil, inguinal lymph node, liver, spleen, kidney, skeletal muscle
(ham), and colon contents (feces), and examined using both real
time RT-PCR and virus isolation (VI) techniques, which are the most
sensitive and specific tools to detect the presence of viral
nucleic acid and live virus, respectively.
[0219] Results:
[0220] Tissues outside the respiratory tract were found to be
negative by VI at 5 days post infection. Only respiratory tract
samples were positive by both methods (real time RT-PCR and VI).
The inguinal lymph node from one pig and serum from two pigs were
positive by real time RTPCR. However, lymph node and serum samples
from all pigs were negative by VI. By contrast, all day 5 post
infection nasal swabs and lung lavage fluids were positive by real
time RT-PCR and VI, and lung tissue homogenates from all four pigs
were positive by real time RT-PCR and 2/4 samples positive by
VI.
[0221] Conclusion:
[0222] Live 2009 A/H1N 1 Influenza A Virus was only detected in the
respiratory tract of infected pigs and the virus does not appear to
spread and replicate in other tissues based on the day 5 post
infection samples.
[0223] In addition, while not bound by theory, the efficacy of
porcine enzymes may be enhanced by vaccinating pancreatic porcine
donors with Trivalent inactivated influenza vaccine (TIV) or Live
attenuated influenza vaccine LAIV. Current test results suggest
that vaccinated pigs demonstrate a pre-existing immunity to certain
currently circulating H1N1 SW strains may protect against an
outbreak virus.
[0224] Experiment 11
[0225] Recent Results from Studies with the 2009 A/H1N1 Influenza A
VirusProject 1: Serologic cross-reactivity of serum samples from
U.S pigs against the new 2009H1N1 influenza virus.
[0226] Purpose of Study:
[0227] An important concern is to address whether U.S commercial
swine herds are susceptible to the 2009 A/H1N1 influenza viruses
isolated from persons in California, New York, and Mexico.
[0228] Experiment:
[0229] Three 2009 A/H1N1 influenza A viruses isolated from persons
in 2009 in California A/CA/04/2009), New York (A/NY/18/2009), and
Mexico (A/Mexico/4108/2009) were obtained from the Centers for
Disease Control and Prevention (CDC) and grown in vitro (i.e., in a
permissive cell line).
[0230] A standard hemagglutination inhibition (HI) test was used to
investigate antigenic relatedness between these three 2009 A/H1N1
influenza A viruses and 19 H1 Swine Influenza Virus (SIV) strains
known to be circulating in U.S. swine herds or with SW strains used
for five licensed U.S H1N1 SW vaccines. Antigenic relatedness would
be predicted on the basis of how well these antisera could inhibit
the three 2009 A/H1N1 influenza A viruses from agglutinating
(clumping) red blood cells. This test indicates the presence of
antibodies that prevent the influenza virus from attaching to red
blood cells and is therefore indicative that the animal may have
protective antibodies. The CDC and USDA-APHIS-Center for Veterinary
Biologics report an 8-fold or greater reduction in HI titer a
significant reduction in cross reactivity between virus
hemagglutinin variants.
[0231] Thirty-eight serum samples from pigs vaccinated with 19 H1
SIV isolated from U.S commercial swine operations between 1999-2008
(NADC H1 serum reference panel) were tested in the standard H1
test. The 19 H1 SIV in the NADC H1 serum reference panel used in
this study represent all four phylogenetic (genetically
characterized) clusters (.alpha., .beta., .gamma., and .delta.) of
all the endemic H1 swine influenza viruses known to circulate in
the U.S. An additional 14 serum samples from pigs vaccinated with
five different commercial products used to vaccinate pigs against
H1 swine influenza viruses in the U.S were tested by the standard
H1 test.
[0232] Results: Eleven of the thirty-eight serum samples from pigs
inoculated with U.S H1N1 SW had a measurable H1 titer against the
A/CA/04/2009 H1N1 influenza virus. The same experiment with the
A/NY/18/2009 H1N1 influenza virus had similar results. In contrast,
twenty two of the thirty-eight serum samples from pigs inoculated
with U.S H1 SIV had a measurable H1 titer against the
(A/Mexico/4108/2009) H1N1 influenza virus. Serologic
cross-reactivity with anti-sera from 5 commercially-available SW H1
vaccines was additionally assessed by H1 with the three 2009 A/H1N1
strains. Cross reactivity was consistently low between the vaccine
antisera and all 2009 A/H1N1 novel strains tested, although titers
were slightly higher with the isolate from Mexico. This suggests
that currently available vaccines may provide only limited
protection against challenge with the novel H1N1.
[0233] Conclusion:
[0234] Results of this experiment suggest that pre-existing
immunity induced by swine influenza viruses circulating in the U.S
swine herd may not protect pigs against the new 2009 A/H1N1
influenza viruses presently circulating in people. Importantly,
vaccines currently used to protect pigs in U.S swine operations
against swine influenza virus may not be effective against the new
2009 H1N1 influenza viruses.
[0235] Limited cross-reactivity of serum samples from the NADC H1
SIV antiserum reference panel or sera from pigs vaccinated with
commercial vaccines was demonstrated against the 2009 A/H1N1
influenza virus (A/CA/04/2009) isolated in California as measured
by a standard H1 test. A second 2009 A/H1N1 strain from New York,
A/NY/18/2009, was also used with the NADC H1 antiserum reference
panel with very similar results to A/CA/04/2009. However, a third
strain, A/Mexico/4108/2009, demonstrated broader cross-reactivity
with the NADC H1 antiserum reference panel. This was especially
apparent in the Hly phylogenetic cluster. The cross-reactivity with
the Hly phylogenetic cluster is important since this is the genetic
group in which the HA from the 2009 A/H1N1 originated. This would
suggest that pre-existing immunity to certain currently circulating
H1N1 SIV strains may protect against the outbreak virus. However,
the differences between the novel H1N1 isolates suggest that there
may be biologic variation in host and/or virus properties
responsible for the variation in serologic crossreactivity.
[0236] It remains unknown whether this variation would have any
effect on protection from live challenge in pigs from circulating
strains of the 2009 A/H1N1 from the human population. Serologic
cross-reactivity with anti-sera from 5 vaccines was also assessed
by H1 with the three 2009 A/H1N1 strains. Cross reactivity was
consistently low between the vaccine antisera and all 2009 A/H1N1
novel strains, although titers were slightly higher with the
isolate from Mexico. This suggests that currently available
vaccines may provide only limited protection against challenge with
the 2009 H1N1.
EXAMPLES
[0237] Proposed Experiments
[0238] Experiment 1--In Vitro Testing
[0239] Reactivity of A/H1N1 influenza A virus to uncoated
pancreatic porcine enzyme dilutions in a pH neutral medium.
[0240] Purpose of Study
[0241] To address the efficacy of uncoated pancreatic porcine
enzymes in eradicating or inhibiting the spread of various strains
of A/H1N1 influenza virus by measuring the degradation in viral RNA
integrity following exposure to an enzymatic preparation.
[0242] Experiment:
[0243] Influenza virus type A, H1N1, isolated from persons in
various time periods and geographic locations will be obtained from
the Centers for Disease Control and Prevention (CDC) or World
Health Organization (WHO) and grown in vitro (i.e., in a permissive
cell line). Appropriate viral containment facilities will be
employed.
[0244] A/H1N1 virus will be grown in culture using infection into
an appropriate cell host such as MDCK anchorage dependant cells
cultures or CACO-2 cell lines. A/H1N1 viral particles shed into the
culture medium will then be collected and the viral particle
concentration determined using a real-time Reverse Transcriptase
Polymerase Chain Reaction (rRT-PCR) based assay for a/H1N1 specific
RNA. Porcine Enzyme Concentrate will then be added to a mixture of
the viral particles prepared in standard cell culture growth media
used with either of the above two named cell lines. Various
dilutions will be prepared including 1:25, 1:50, 1:100, 1:200.
Materials will be incubated for a period of time ranging from 30
minutes to 6 hours at 37 C. Virus particles will then be separated
from the Enzyme preparation, reconstituted in growth medium and
plated onto the CACO-2 or MDCK cell lines. Cells will be allowed to
grow and titer of released viral particles over a period of 3 days
will be determined. Controls will be prepared by exposing A/H1N1
viral particles to phosphate buffered saline solution at 37 C for
identical times. Control particles will then be prepared and plated
as per experimentally treated virus cultures.
[0245] Samples at 12 hour periods over the course of 3 days will be
assayed using laboratory diagnostic testing for the presence of
influenza viruses in the specimens. In addition to utilization of a
Real-time Reverse Transcriptase Polymerase Chain Reaction based
assay (rRT-PCR) that has been shown capable of detecting the A/H1N1
specific RNA, other tests may be used including direct antigen
detection tests such as ELISA based assays for viral antigens and
virus isolation in cell culture may be used.
[0246] Other rRT-PCR assays such as laboratory developed tests, not
approved by FDA, may also be able to detect novel influenza A
(H1N1) viruses. Public health laboratories in the U.S. are
currently able to perform the CDC rRT-PCR Swine Flu Panel
assay.
[0247] Anticipated Results:
[0248] Replication of A/H1N1 in a permissive cell line should be
inhibited (static), reduced or eradicated (-cidal) by exposure to
the pancreatic porcine enzyme dilutions as compared to control
samples.
[0249] Experiment 2--In Vivo Testing
[0250] Effectiveness of coated pancreatic porcine enzyme on A/H1N1
influenza A virus symptoms or viral load in humans.
[0251] Purpose of Study
[0252] To address the efficacy of coated pancreatic porcine enzymes
in eradicating or decreasing the symptoms or viral load of A/H1N1
influenza virus.
[0253] Experiment:
[0254] Candidate test subjects are those individuals who exhibit
symptoms of A/H1N1 influenza A viruses and meet other test
inclusion or exclusion criteria will be tested for presence of the
virus using rapid A/H1N1 testing. Test Groups may be selected by
age including infant and elderly, general health including healthy
subjects and immuno-compromised subjects, geographic location, or
other selection/exclusion criteria.
[0255] Rapid influenza diagnostic testing is utilized to identify
candidate test subjects in a clinically relevant time period. Rapid
influenza diagnostic tests (RIDTs) are typically antigen detection
tests that detect influenza viral nucleoprotein antigen. The
present commercially available test can provide results within 30
minutes or less. These assays may be referred to as "point-of care"
tests since CLIA-waived RIDTs (not all RIDTs are CLIA waived) may
be used in facilities with a certificate of waiver or in locations
outside a central laboratory. Commercially available RTDTs can
either: i) detect and distinguish between influenza A and B
viruses; ii) detect both influenza A and B but not distinguish
between influenza A and B viruses; or, iii) detect only influenza A
viruses. None of the currently FDA approved RIDTs can distinguish
between influenza A virus subtypes (e.g. seasonal influenza A
(H3N2) versus seasonal influenza A (H1N1) viruses), and RTDTs
cannot provide any information about antiviral drug
susceptibility.
[0256] Once a positive result is obtained further swabs or test
specimens are obtained from the test subjects for detailed
laboratory analysis to validate positive test subject and isolate
specific viral subtype(s). Laboratory tests may include direct
antigen detection tests, virus isolation in cell culture, or
detection of influenza-specific RNA by real-time reverse
transcriptase-polymerase chain reaction (rRT-PCR).
[0257] Laboratory tests typically differ in their sensitivity and
specificity in detecting influenza viruses as well as in their
commercial availability, the amount of time needed from specimen
collection until results are available, and the tests' ability to
distinguish between different influenza virus types (A versus B)
and influenza A subtypes (e.g. novel H1N1 versus seasonal H1N1
versus seasonal H3N2 viruses). At the present time, there are only
two FDA authorized assays for confirmation of novel influenza A
(H1N1) virus infection, including the CDC rRT-PCR Swine Flu Panel
assay.
[0258] Other rRT-PCR assays such as laboratory developed tests, not
approved by FDA, may also be able to detect novel influenza A
(H1N1) viruses. Public health laboratories in the U.S. are
currently able to perform the CDC rRT-PCR Swine Flu Panel assay
[0259] Stool samples may be collected from the subjects both prior
to the start of clinical testing and periodically during testing.
This may especially important when testing immune-compromised
individuals who may have low levels of chymotrypsin or other
pancreatic secretions in their stool.
[0260] Coated Porcine Enzyme Concentrate will be administered to
test subjects at appropriate doses with each major meal for a
minimum of 3 dosings per day over a period of 7 to 10 days. Swabs
or test specimens and, optionally, stool samples are collected on a
daily basis for subsequent laboratory analysis over a clinically
relevant time period such as 7 to 10 days. Patients will be
observed for progression of symptoms, viral related sequelae and
the presence of viral particles in collected samples over the
course of treatment.
[0261] Placebos arc also administered to a control sub group of
test subjects at the same time intervals and swabs or test
specimens and optionally stool samples are collected on a daily
basis for subsequent laboratory analysis over a clinically relevant
time period such as 7 to 10 days.
[0262] Anticipated Results:
[0263] Administration of coated pancreatic porcine enzymes to test
subjects positive for A/H1N1 should decrease the severity and/or
duration of associated symptoms. Additionally, compared to placebo
treated patients, there should be an accelerated decrease in
detectable virus obtained from physiological samples.
[0264] The foregoing description of the embodiments of the
disclosure has been presented for the purposes of illustration and
description. It is not intended to be exhaustive or to limit the
disclosure to the precise form disclosed. Many modifications and
variations are possible in light of this disclosure. It is intended
that the scope of the disclosure be limited not by this detailed
description, but rather by the claims appended hereto.
* * * * *