U.S. patent application number 17/152103 was filed with the patent office on 2021-10-21 for sickle cell disease treatment utilizing omega-3 fatty acids.
This patent application is currently assigned to MICELLE BIOPHARMA, INC.. The applicant listed for this patent is MICELLE BIOPHARMA, INC.. Invention is credited to Ahmed Abd Almalik Ahmed Mohammed Daak, Glynis Daniel-Archibald, Miguel Lopez-Toledano, Frederick D. Sancilio, Thorsteinn Thorsteinsson.
Application Number | 20210322359 17/152103 |
Document ID | / |
Family ID | 1000005692890 |
Filed Date | 2021-10-21 |
United States Patent
Application |
20210322359 |
Kind Code |
A1 |
Sancilio; Frederick D. ; et
al. |
October 21, 2021 |
SICKLE CELL DISEASE TREATMENT UTILIZING OMEGA-3 FATTY ACIDS
Abstract
Described herein are compositions including at least one omega-3
fatty acid (either in the triglyceride, ester or free fatty acid
ester form) and at least one surface active agent; wherein the
compositions form micelles when in contact with an aqueous medium.
Also provided are methods of administering to a subject a
composition including at least one omega-3 fatty acid (either in
the triglyceride, ester or free fatty acid ester form) and at least
one surface active agent, wherein the compositions form micelles
when in contact with an aqueous medium, and the bioavailability of
the omega-3 fatty acid is substantially independent of a food
effect. The compositions are useful for treating certain disease
states which may include (1) malabsorption syndromes, (2) primary
sclerosing cholangitis (PSC), (3) non-alcoholic fatty liver disease
(NAFLD), (4) sickle cell disease (SCD), (5) age-related macular
degeneration (AMD), and (6) neurodegenerative disease, including,
Parkinson's Disease (PD), Alzheimer's Disease (AD), Amyotrophic
Lateral Sclerosis (ALS or Lou Gehrig's Disease), Epilepsy, Bi-polar
Syndrome, traumatic brain injury, peripheral neuropathy, and
Multiple Sclerosis (MS). Described are also various dosage forms
for administering the compositions and use of the compositions in
functional foods. Provided herein are also kits with instructions
for their administration.
Inventors: |
Sancilio; Frederick D.;
(Riviera Beach, FL) ; Thorsteinsson; Thorsteinn;
(Riviera Beach, FL) ; Daniel-Archibald; Glynis;
(Riviera Beach, FL) ; Lopez-Toledano; Miguel;
(Riviera Beach, FL) ; Daak; Ahmed Abd Almalik Ahmed
Mohammed; (Riviera Beach, FL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MICELLE BIOPHARMA, INC. |
RIVIERA BEACH |
FL |
US |
|
|
Assignee: |
MICELLE BIOPHARMA, INC.
RIVIERA BEACH
FL
|
Family ID: |
1000005692890 |
Appl. No.: |
17/152103 |
Filed: |
January 19, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16655643 |
Oct 17, 2019 |
10894027 |
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17152103 |
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15605972 |
May 26, 2017 |
10898458 |
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16655643 |
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15180430 |
Jun 13, 2016 |
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15605972 |
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PCT/US15/54933 |
Oct 9, 2015 |
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15180430 |
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62062646 |
Oct 10, 2014 |
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62062654 |
Oct 10, 2014 |
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62062643 |
Oct 10, 2014 |
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62062638 |
Oct 10, 2014 |
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62062634 |
Oct 10, 2014 |
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62062651 |
Oct 10, 2014 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/355 20130101;
A61K 9/1075 20130101; A61K 47/10 20130101; A61K 9/4858 20130101;
A61K 31/05 20130101; A61K 31/202 20130101; A61K 47/26 20130101;
A61K 31/7024 20130101; A61K 31/353 20130101; A61K 31/201 20130101;
A61K 31/232 20130101 |
International
Class: |
A61K 31/232 20060101
A61K031/232; A61K 9/107 20060101 A61K009/107; A61K 9/48 20060101
A61K009/48; A61K 31/05 20060101 A61K031/05; A61K 31/201 20060101
A61K031/201; A61K 31/202 20060101 A61K031/202; A61K 31/353 20060101
A61K031/353; A61K 31/355 20060101 A61K031/355; A61K 31/7024
20060101 A61K031/7024; A61K 47/10 20060101 A61K047/10; A61K 47/26
20060101 A61K047/26 |
Claims
1. A method of treating or preventing sickle cell disease or
disease symptoms, comprising administering an effective amount of a
composition to a subject in need thereof, the composition
comprising: at least 90% pure docosahexaenoic acid (DHA) and at
least one surface active agent, wherein said at least one surface
active agent comprises a combination of at least one polysorbate
and at least one poloxamer, wherein said at least one polysorbate
comprises from about 30.0% wt/wt to about 32.0%) wt/wt of said
composition, wherein said at least one poloxamer comprises from
about 0.60% wt/wt to about 0.80%) wt/wt of said composition,
wherein said at least one polysorbate is polysorbate 80, and
wherein said at least one poloxamer is poloxamer 237.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 16/655,643 filed on Oct. 17, 2019.
[0002] U.S. patent application Ser. No. 16/655,643 is a
continuation of U.S. patent application Ser. No. 15/605,972 filed
May 26, 2017.
[0003] U.S. patent application Ser. No. 15/605,972 is a
continuation of U.S. patent application Ser. No. 15/180,430 filed
Jun. 13, 2016.
[0004] U.S. patent application Ser. No. 15/180,430 is a
continuation of International Patent Application No.
PCT/US15/54933, filed Oct. 9, 2015, which claims priority from U.S.
Provisional Patent Application Nos. 62/062,638 filed Oct. 10, 2014;
62/062,643 filed Oct. 10, 2014; 62/062,646 filed Oct. 10, 2014;
62/062,652 filed Oct. 10, 2014; 62/062,634 filed Oct. 10, 2014; and
62/062,651 filed Oct. 10, 2014.
FIELD OF THE INVENTION
[0005] This invention relates to compositions including fatty
acids, either in the triglyceride, ester, or free fatty acid forms,
in combination with at least one surface active agent, and to the
use of such compositions for the treatment of patients afflicted
with various disease states. More particularly, this invention
relates to self-micellizing compositions including at least one
omega-3 fatty acid, which may be selected from the group consisting
of hexadecatrienoic acid, .alpha.-linolenic acid, stearidonic acid,
eicosatrienoic acid, eicosapentaenoic acid, heneicosapentaenoic
acid, the omega-3 isomer of docosapentaenoic acid, also known as
clupanodonic acid, docosahexaenoic acid, tetracosapentaenoic acid,
and tetracosahexaenoic acid, in combination with a surface active
agent composition effective to spontaneously form micelles with
said fatty acids upon contact with an aqueous media. In certain
embodiments, the self-micellizing compositions may further contain
an omega-6 fatty acid (either in the triglyceride, ester, or free
fatty acid form), for example arachidonic acid (ARA), linoleic acid
(LA), gamma-linolenic acid (GLA) or the omega-6 isomer of DPA, also
known as osbond acid. Most particularly, this invention relates to
the treatment of various disease states by administering to a
patient in need thereof the aforementioned compositions or
pre-formed micelles produced therefrom.
[0006] In certain embodiments, the disease states may include (1)
malabsorption syndromes, such as short bowel syndromes, which
encompass a number of different clinical entities that may result
in chronic diarrhea, abdominal distention, and failure to thrive;
(2) primary sclerosing cholangitis (PSC), which is a chronic
cholestatic liver disease characterized by progressive inflammatory
and fibrotic destruction of the intrahepatic and/or extrahepatic
bile ducts; (3) non-alcoholic fatty liver disease (NAFLD), which is
characterized by increased hepatic fat accumulation in individuals
not consuming excessive alcohol and represents a spectrum of
disease ranging from `simple` steatosis to non-alcoholic
steatohepatitis; (4) sickle cell disease, which is a group of blood
disorders that affects hemoglobin, the molecule that delivers
oxygen throughout the body via red blood cells; (5) age-related
macular degeneration (AMD), which is a condition results in a loss
of vision in the center of the visual field (the macula); and (6)
neurodegenerative disease, which is an umbrella term for a range of
conditions, which primarily affect the neurons in the human brain.
Examples of neurodegenerative diseases contemplated for treatment
by the present invention include, but are not limited to,
Parkinson's Disease (PD), Alzheimer's Disease (AD), Amyotrophic
Lateral Sclerosis (ALS or Lou Gehrig's Disease), Epilepsy, Bi-polar
Syndrome, traumatic brain injury, peripheral neuropathy, and
Multiple Sclerosis (MS).
BACKGROUND
[0007] Malabsorption Syndromes
[0008] Malabsorption syndromes encompass a number of different
clinical entities that result in chronic diarrhea, abdominal
distention, and failure to thrive. Clinical malabsorption can be
broken down into several distinct conditions, both congenital and
acquired, that affect one or more of the different steps in the
intestinal hydrolysis and subsequent transport of nutrients.
[0009] Malabsorption is caused by a disorder in the intestinal
processes of digestion and/or transport of nutrients across the
intestinal mucosa into the systemic circulation. Either a
congenital abnormality in the digestive or absorptive processes or,
more commonly, a secondarily acquired disorder of such processes
may result in malabsorption.
[0010] Neonates, young infants and adolescent humans with
malabsorption syndromes, in particular fat malabsorption syndromes,
are at particularly high risk for chronic diarrhea and malnutrition
due to insufficient energy intake of high-energy value dietary
lipids and essential lipids, which constitute building blocks for
normal function of multiple body systems.
[0011] In some humans, exocrine pancreatic insufficiency is the
principal condition that results in severe fat malabsorption.
Pancreatitis, pancreatic cancer, pancreatic resection, cystic
fibrosis, Shwachman-Diamond syndrome, Johnson-Blizzard syndrome,
and Pearson syndrome can all result in pancreatic insufficiency.
Significant obstructive biliary or cholestatic liver disease or
extensive intestinal mucosal disease, such as occurs in celiac
disease, may also result in severe steatorrhea.
[0012] One particular malabsorption syndrome is Short-Bowel
Syndrome (SBS)--a disorder clinically defined by a failure to
properly absorb nutrients (malabsorption), frequently accompanied
by diarrhea, steatorrhea, fluid and electrolyte disturbances,
edema, dehydration and malnutrition. Short-bowel syndrome has
numerous causes, both congenital and acquired, but a common
etiologic factor is the functional or anatomic loss of extensive
segments of small intestine leading to a severe decrease in
intestinal absorptive capacity. Adult human intestines have a
normal length range of 260-88 cm. If disease, accident, or any
other cause results in a loss of 50% of the small intestine or
leaves less than 200 cm of viable length, a patient is at high risk
of SBS.
[0013] The link between small intestine loss and short-bowel
syndrome is simple, losing large amounts of the small intestine
compromises the digestive and absorptive processes. Adequate
digestion and absorption cannot take place and proper nutritional
status cannot be maintained without supportive care, frequently
including short term and long-term parenteral nutrition. In adults,
SBS is commonly caused by radiation enteritis, mesenteric vascular
accidents, trauma, and recurrent intestinal obstruction. In
children, the most common causes are necrotizing enterocolitis,
intestinal atresias, and intestinal volvulus. Other causes of SBS
include congenital short small bowel, gastroschisis, and meconium
peritonitis.
[0014] Whether or not a patient who has lost a significant amount
of small intestine will develop SBS depends on a number of factors.
Important cofactors that help to determine whether the syndrome
will develop or not include the premorbid length of the small
intestine, how much intestine is lost, the age of the patient, the
remaining length of small intestine and colon, the functional
quality of the residual bowel, and the presence or absence of the
ileocecal valve.
[0015] Treatment of malabsorption syndromes, such as SBS, is mainly
aimed at supplying the nutrients and vitamins that patients lack.
It may include (a) a high-calorie diet that includes vitamins,
minerals, carbohydrates, proteins, and fats; (b) injections of
vitamins and minerals; (c) administration of drugs that slow down
the normal movement of the small intestine; and (d) feeding through
a vein (Total Parenteral Nutrition (TPN)).
[0016] Most SBS patients are initially fed with TPN. In many
patients, intestinal adaptation, alone or in combination with
dietary modifications, allows weaning from TPN. Unfortunately, many
patients cannot be weaned from TPN.
[0017] Patients on long-term TPN frequently experience serious
metabolic complications. The most common complications are hepatic
and biliary disorders manifested by steatosis, fibrosis and
cholestasis. These disorders can progress to fulminant liver
failure. Prior to 2006, advanced liver disease was the most common
cause of death in SBS patients. In infants with SBS, TPN-associated
liver disease was estimated to occur in 40-60% of those receiving
long-term TPN. This is largely due to the fact that infants are
currently administered an intravenous solution of DHA, leading to
liver injury, Mortality in infants who develop cholestasis is
estimated to be as high as 80%. In accordance with the present
invention, it is possible to significantly decrease this
treatment-related morbidity by administering substantially pure DHA
in combination with at least one surface active agent, as described
herein, effective to spontaneously form a plurality of stable
micelles having a particle size within a range of about 1 .mu.m to
about 10 .mu.m upon contact with an aqueous medium, or
alternatively, by administering pre-formed micelles produced
therefrom, because the DHA avoids the first pass effect, the fatty
acid is in the less toxic chylomicron form in blood, and there is
no IV fistula issue.
[0018] Patients with malabsorption syndromes require products that
safely and effectively deliver all essential nutrients. Many
consider TPN-associated liver disease (PNALD) to be caused in part
by the intravenous lipid emulsions used to provide fat calories in
the PN formulation, although the etiology is unclear. It has been
suggested that PNALD may be caused by inflammation that is related
to the type of intravenous lipid emulsion used. These fat emulsions
cannot be eliminated from PN because to do so would lead to the
development of essential fatty acid deficiency and its concomitant
complications. Recent data has shown that the replacement of
omega-6 soybean oil lipid emulsions with those comprised primarily
of omega-3 fatty acids may lead to reduction or avoidance of PNALD
without predisposing the patient to essential fatty acid
deficiency.
[0019] Given the limited treatment options currently available to
patients with fat malabsorption syndromes and the associated side
effects of these treatments, there is a need for alternative
medications that can treat fat malabsorption syndromes. Ideally,
such medications should provide the necessary and the essential
fatty acids and fat calories for proper growth, development and
maintenance of bodily functions in humans of all ages, but
especially in neonates, and reduce or even avoid PNALD.
[0020] Primary Sclerosing Cholangitis
[0021] Primary sclerosing cholangitis (PSC) is a chronic
cholestatic liver disease characterized by progressive inflammatory
and fibrotic destruction of the intrahepatic and/or extrahepatic
bile ducts. As the bile ducts become inflamed and narrow, bile
backs into the liver causing liver cells to become inflamed. Over
time this inflammation decreases blood flow within the liver,
increasing pressure in the portal vein. As the disease progresses,
liver cells die and are replaced with scar tissue.
[0022] The etiology of PSC is unknown, although, the association of
PSC with inflammatory bowel disease, autoimmune diseases, and a
host of other humoral and cellular immune abnormalities make an
immunopathogenic mechanism likely.
[0023] Although the initial diagnosis of PSC is usually made within
the fourth decade of life with a mean age of 40 years of age in men
and 45 years in women, PSC is increasingly diagnosed in infants (in
neonates, PSC is referred to as Neonatal Sclerosing Cholangitis
(NSC)), children, and adolescents. PSC predominantly affects males.
The diagnosis is based on characteristic findings in combination
with clinical, biochemical and histological features. Many patients
are asymptomatic at presentation, but symptoms develop over time.
Symptoms include itch, lethargy, steatorrhea and vitamin
deficiencies, metabolic bone disease, bleeding peristomal varices,
bacterial cholangitis, dominant biliary strictures, and in advanced
cases cholangiocarcinoma.
[0024] There is increasing evidence of a genetic predisposition to
PSC, but the exact mechanism of susceptibility has not been well
defined.
[0025] Immunosuppressants, chelators, and steroids are frequently
used to help control the disease process, but have not shown any
significant benefit. Ursodeoxycholic acid has been studied and may
improve the liver function profile, but a high number of adverse
events are associated with this treatment and there is debate about
its effectiveness.
[0026] Genotype and phenotype analyses have shown that there is an
increased prevalence of cystic fibrosis transmembrane conductance
regulator protein (CFTR) abnormalities in adults with PSC. The CFTR
protein functions as a channel for the movement of ions in and out
of the epithelial cells lining exocrine glands. CFTR dysfunction is
associated with (1) decreased chloride secretions into the bile
canaliculi with subsequent decrease in osmotic extrusion of water
into the lumen, resulting in hyperconcentration and acidification
of bile leading to obstruction of intrahepatic bile ductules,
secondary inflammation and eventual focal biliary cirrhosis; (2)
innate immune defects and leads to an excessive host inflammatory
response; (3) fatty acid alterations, specifically a decrease in
docosahexaenoic acid (DHA) and an increase in arachidonic acid
(AA). As CFTR dysfunction seems to be a contributing factor in the
development of primary sclerosing cholangitis, targeting the
correction of the fatty acid abnormalities and associated defects
in immune defense and inflammatory responses seems to be a
promising path in the treatment of PSC.
[0027] Omega-3 fatty acids are known to have anti-inflammatory
effects. DHA in particular is known to be an important regulator of
inflammation. It has been recently discovered that resolvins and
protectins, two types of lipid mediators derived from DHA, modulate
the inflammatory response by actively promoting the resolution of
inflammation. DHA has been shown to correct fatty acid
abnormalities and reverse the development of bile duct injury in
CFTR.sup.-/- knockout mice. Recent data from a 12-month, open-label
pilot study (n=23) to evaluate the safety and effectiveness of DHA
for the treatment of PSC showed an increase in serum DHA levels and
a significant decline in alkaline phosphatase in patients with
PSC.
[0028] Currently there are no effective medical therapies for PSC
that result in significant long-term improvement in outcome. Liver
transplantation is the only life-extending therapy for patients
with end-stage disease, with all of its associated risks, costs and
complications. The median time from diagnosis to death or liver
transplantation is 9-18 years. While different forms of medical
treatment have been tried, there are no established, effective
therapies that result in significant long-term improvements in
outcome other than liver transplantation. Most treatments focus on
monitoring liver function, managing symptoms and, when possible,
temporarily opening blocked bile ducts. These therapies do not
target the excessive inflammation and subsequent biliary tract
fibrosis associated with the disease. Thus, there is clearly an
unmet need for therapies, especially those that use omega-3 fatty
acids, that can effectively target the inflammation and fibrosis
associated with PSC and that therefore may prevent or limit disease
progression.
[0029] Non-Alcoholic Fatty Liver Disease and Non-Alcoholic
Steatohepatitis
[0030] Non-Alcoholic Fatty Liver Disease (NAFLD) is a common, often
"silent" liver disease and resembles alcoholic liver disease, but
occurs in people who drink little or no alcohol. The major feature
of NAFLD is excess fat in the liver, along with inflammation and
damage. Most people with this disease feel well and are not aware
that they have a liver problem. Nevertheless, the condition can be
severe and can lead to cirrhosis, in which the liver is permanently
damaged and scarred and no longer able to function properly.
[0031] Non-alcoholic fatty liver disease (NAFLD) is characterized
by increased hepatic fat accumulation in individuals not consuming
excessive alcohol and represents a spectrum of disease ranging from
`simple` steatosis to non-alcoholic steatohepatitis. NAFLD is
associated with the metabolic syndrome and is defined by the
presence of .gtoreq.5% hepatic steatosis. Risk factors for the
development of NAFLD include central obesity, type II diabetes,
dyslipidemia, and hypertension. In Western populations, the
prevalence of NAFLD may exceed 30%, and can be as high as 88% in
the obese. NAFLD is the most common liver disease in children.
Given the increasing prevalence and incidence of these conditions,
the global burden of NAFLD is expected to increase.
[0032] Currently, NASH and NAFLD are under diagnosed due to poor
disease awareness, the insufficiency of non-invasive diagnostic
tools and the lack of effective approved therapies. As a confirmed
diagnosis of NASH currently requires a liver biopsy, patients are
often diagnosed after a blood test demonstrating elevated levels of
liver enzymes, alanine aminotransferase (ALT), and aspartate
aminotransferase (AST).
[0033] The primary treatment for NAFLD is weight loss by lifestyle
therapy involving diet and exercise. Weight loss has been shown to
improve liver enzymes, decrease plasma triglycerides and improve
liver fatness. Bariatric surgery is an extreme option for reducing
weight and the resulting improvements in liver pathology. However,
bariatric surgery is not feasible for the large number of patients
having this disease.
[0034] Treatments with insulin sensitizers, hypolipidemics and
vitamin E have been shown to be ineffective due to poor patient
compliance, associated weight gain, and side-effects. Although
current evidence from available randomized control trials suggests
that while thiazolidinediones and vitamin E are effective in
reducing liver fat, there are serious safety concerns about
long-term use of these agents.
[0035] Non-alcoholic steatohepatitis (NASH) is a fibrotic
manifestation, which progresses from NAFLD. Like NAFLD, NASH is
usually a silent disease with few or no symptoms and occurs in
people who drink little or no alcohol. The major feature in NASH is
fat in the liver, along with inflammation and damage. Most people
with NASH feel well and are not aware that they have a liver
problem. Nevertheless, NASH can be severe and can predispose the
individual to hepatic fibrosis, cirrhosis, and subsequent end-stage
liver disease and hepatocellular carcinoma. Not every person with
NASH develops cirrhosis, but once serious scarring or cirrhosis is
present, few treatments can halt the progression. A person with
cirrhosis experiences fluid retention, muscle wasting, bleeding
from the intestines, and liver failure. Liver transplantation is
the only treatment for advanced cirrhosis with liver failure, and
transplantation is increasingly performed in people with NASH. NASH
ranks as one of the major causes of cirrhosis in the United States,
behind hepatitis C and alcoholic liver disease.
[0036] Despite the rapidly increasing incidence of NASH, there are
no therapies currently approved for the treatment of this common
liver disorder. The NASH market has a significant unmet need for
pharmacological options that are effective and well tolerated.
Current options for managing patients with NASH are suboptimal and
primarily rely on changes in lifestyle to reduce weight, off-label
pharmacotherapy and bariatric surgery for weight loss. Weight loss
is the first recommendation for NASH patients and is associated
with a significant improvement in steatosis and overall severity of
NASH. However, such improvement is only statistically significant
when patients are able to reduce more than 7% of their body weight
over a sustained period of 48 weeks, which occurs in less than 50%
of NASH patients. Products utilized off label in the management of
NASH comorbidities include vitamin E, insulin sensitizers such as
metformin and pioglitazone, which are used for diabetic patients,
and anti-hyperlipidemic agents, pentoxifylline, and ursodiol.
High-dose vitamin E has been shown in a clinical study of
non-diabetic patients to reduce inflammation but not fibrosis.
Vitamin E is not recommended for NASH patients with type-2 diabetes
due to lack of data, therefore the use of vitamin E is limited.
While other off-label pharmacotherapies demonstrate inconsistent
benefits or are associated with significant side effects. Bariatric
surgery is believed to impact NASH through dramatic weight loss,
but it has significant complications and drawbacks. These include a
host of perioperative risk factors, the need to adhere to
post-surgical diet and nutritional regimens and high costs. A
relatively small number of these procedures are performed annually
on NASH patients compared to the overall NASH population, which we
believe is due to the complications and drawbacks of bariatric
surgery relative to NASH patient numbers. We believe widespread
increased adoption of bariatric surgery for NASH is impractical
based on cost and the large number of patients who would require
it. In addition, some retrospective and prospective studies have
indicated that the procedure may worsen fibrosis. Liver transplant
is a last resort for life-threatening complications progressing
from NASH. NASH is currently the third most common reason for liver
transplants in United States and is projected to surpass
alcohol-based cirrhosis and viral hepatitis to become the leading
indication for liver transplant by 2020. The availability of liver
donors is extremely limited and the cost of a liver transplant is a
significant economic burden, with an estimated cost per procedure
of approximately $577,000. Studies have demonstrated that
approximately 22% of patients do not survive the five-year period
post-transplant. Therefore, a significant unmet need exists for
alternative treatments for NAFLD and NASH.
[0037] While NAFLD occurs in people of all ages, NASH most often
occurs in people who are middle aged and overweight or obese.
Affected individuals may also have elevated levels of blood lipids
(such as cholesterol and triglycerides) and many have diabetes.
NASH affects 2 to 5 percent of Americans. Both NASH and NAFLD are
becoming more common, possibly because of the greater number of
Americans with obesity. In the past 10 years, the rate of obesity
has doubled in adults and tripled in children. Obesity also
contributes to diabetes and high blood cholesterol, which can
further complicate the health of a patient with NASH.
[0038] There is no current approved treatment for NASH or NAFLD.
Accordingly, medications for the treatment of NAFLD and/or NASH
represent a substantial unmet need.
[0039] Sickle Cell Diseases
[0040] Sickle cell disease, also known as sickle-cell anaemia (SCA)
and drepanocytosis, is a group of hereditary blood disorders that
affects hemoglobin, the molecule that delivers oxygen throughout
the body via red blood cells. Sickle cell disease is caused by
mutations in the HbA gene and is inherited in an autosomal
recessive pattern (a single point mutation in the sixth codon of
the .beta.-globin gene). There are four common genotypes associated
with SCD--hemoglobin SS (HbSS), HbS beta0-thalassemia, hemoglobin
SC (HbSC), and HbS beta+-thalassemia.
[0041] In individuals with two mutated genes, the resultant
abnormal hemoglobin S polymerizes under low oxygen tension and
causes abnormal red blood cells. Because of the polymerization,
these cells frequently distort into a rigid sickle, or crescent,
shape, in contrast to disc-shaped normal red blood cells. A person
with a single abnormal copy does not experience symptoms and is
said to have sickle-cell trait. A person with sickle cell trait
inherits one normal allele and one abnormal allele encoding
hemoglobin S (hemoglobin genotype AS). Sickle cell trait is
generally regarded as a benign condition, although certain
individuals with sickle cell trait may develop complications.
[0042] Sickle cell disease is a serious disease that significantly
compromises the quality of patients' lives and reduces life
expectancy significantly. Signs and symptoms of sickle cell disease
usually begin in early childhood. The severity of symptoms varies
from person to person and it has been postulated that clinical
manifestations result from complex combinations of genetic,
cellular and environmental factors. Some people have mild symptoms,
while others are frequently hospitalized for more serious
complications.
[0043] Sickle cells have a shorter lifespan than normal red blood
cells. Sickle cells also deliver less oxygen to the body's tissues.
As a result, patents with sickle cell disease frequently develop
anemia. Sickle cell anemia is a common manifestation of sickle cell
disease. Patients with anemia experience fatigue, weakness,
shortness of breath, dizziness, headaches, and coldness in the
hands and feet. Anemia can also cause delayed growth and
development in children. The rapid breakdown of red blood cells may
also cause jaundice.
[0044] Sickle cells are rigid and prone to adhesive interactions
with each other, leukocytes, platelets, plasma and vessel walls.
These adhesive interactions lead to vaso-occlusion in small blood
vessels, with the sickle cells either intact or in pieces.
Vaso-occlusion cuts down even more on the amount of oxygen flowing
to body tissues. Acute vaso-occlusive events are known as crises.
Crises can last from hours to days. Some patients have one episode
every few years while others have many episodes each year. Crises
can be severe enough to require hospitalization and can be fatal.
Vaso-occlusive events in sickle cell disease are believed to be
influenced by multiple factors, including leucocyte adhesion
molecules, inflammatory factors, endothelial cell interactions,
haemorheology and erythrocyte heterogeneity.
[0045] Three serious symptoms often accompany crises; pain, organ
damage and infection. It is common in the relevant literature for
researchers to also refer to "pain crisis" and "hemolytic (or
"haemolytic") crisis" (rapidly evolving anemia, leukocytosis,
jaundice and fever) and "fatty acid membrane abnormality" fatty as
meaningful clinical endpoints.
[0046] Deprivation of oxygen-rich blood is especially deleterious
to the lungs, kidneys, spleen, and brain. A particularly serious
complication of sickle cell disease is pulmonary hypertension
linked to blockages in the blood vessels that supply the lungs.
Pulmonary hypertension occurs in about one-third of adults with
sickle cell disease and can lead to heart failure.
[0047] Other serious consequences of the blocked blood vessels are
strokes, ulcers of the lower extremities, impaired vision and
priapism. Blockage of the blood vessels supplying the spleen may
lead to failure of that organ, which results in serious infectious
conditions such as osteomyelitis, cholecystitis, pneumonia and
urinary tract infection.
[0048] As a result of the vaso-occlusion and organ damage, sickle
cell patients are often in a near-continuous state of inflammation.
They have elevated states of markers of inflammation such as
C-reactive protein, fibrinogen and interleukin. Sickle cell
patients also often have near continuous thrombotic activity, low
level most of the time but spiking during crises.
[0049] There are numerous procedures performed to manage
complications of sickle cell disease. These include blood
transfusions, dialysis or kidney transplants, gallbladder removal,
hip replacements for avascular necrosis, eye surgeries and wound
care for leg ulcers. Bone marrow or stem cell transplants can cure
sickle cell anemia, but are not an option for most patients because
of a lack of well-matched stem cell donors. Ultimately, sickle cell
disease causes multi-organ dysfunction and early death in affected
individuals. Many succumb to complications of chronic organ
dysfunction and eventual organ failure.
[0050] Unfortunately, for decades the root cause of sickle cell
disease was misunderstood. Prior to very recently, the majority
view was that sickle cell disease was merely the result of sickled
red blood cells as a phenotypic manifestation of a single genetic
point mutation. However, recent studies have shown that sickle cell
disease is, in fact, a disease of systemic inflammation, inducing
effects not only on the red blood cells, but also other blood cells
and the vessels themselves. In fact, recent research has shown that
red blood cells do not sickle in sickle cell disease patients until
after a cascade of events occurs which leads to an increased
stickiness of the blood cells and vessels.
[0051] Once the inflammation occurs, the red blood cells sickle,
resulting in a feedback loop cascade, which increases the magnitude
of the inflammation. This increased inflammation is manifested
through the increased presence of arachidonic acid and a decrease
in certain omega-3 acids, such as DHA. Thus, without being bound to
any particular theory, it is believed that restoring the fatty acid
compositions to nearer to normal levels will create a significant
decrease in the inflammation and thus diminish the morbidity of
sickle cell disease.
[0052] Unless a specific condition, disease state or malady
associated with sickle cell disease is referenced herein, as used
herein, a reference to "sickle cell disease" or "sickle cell
condition" or "sickle cell disease condition" refers to each and
all of the disease states and/or effects and/or conditions of
sickle cell disease as described herein or as otherwise known by
the ordinarily skilled artisan in the relevant art. A reference to
"at least one sickle cell condition" refers to one or more of the
conditions related to sickle cell disease as described herein or as
otherwise known to such ordinarily skilled artisan.
[0053] Existing treatments for sickle cell disease are primarily
aimed at relieving symptoms, limiting the number of crises and
lessening complications. People with sickle cell disease need
ongoing treatment, even when not having a crisis.
[0054] Frequently prescribed therapeutic compositions, thought to
be of benefit for sickle cell patients, include pain medications,
antibiotics, folic acid, hydroxyurea, and anti-inflammatory
medications. Pain medicines of all types are heavily prescribed to
address the pain episodes associated with sickle cell disease.
Antibiotics are frequently prescribed to treat the bacterial
infections that are common in sickle cell patients. Sickle cell
patients are often advised to take folic acid supplements because
folic acid aids in the production of new red blood cells.
[0055] Hydroxyurea (tradename DROXIA) is approved to reduce the
number of pain episodes (including chest pain and breathing
problems) and blood transfusions in patients with recurring
moderate to severe crises but is a cytotoxic molecule with
teratogenic and carcinogenic effects. The precise mechanism of
action of hydroxyurea is uncertain. It has been found that besides
the damage caused by sickle cells themselves, the inflammatory
response that occurs in sickle cell disease patients could
potentially play a significant role in the occurrence of painful
episodes or pain crises. Sickle cell patients experience relief of
symptoms when treated with a variety of anti-inflammatory
agents.
[0056] As early as 1991, it was suggested that omega-3 fatty acids
decrease the hemolysis of mammalian red blood cells. It also has
been found that sickle cell patients have abnormal blood fatty
acids. These findings have led to the hypothesis that omega-3 fatty
acids may be useful in the treatment of sickle cell disease. As
early as 2001, small human clinical trials showed that omega-3
fatty acids could reduce pain episodes in sickle cell patients,
perhaps by reducing prothrombotic activity. Other studies have
shown that omega-3 fatty acids can increase hemoglobin levels, and
reduce pain episodes, vaso-occlusive episodes, anemia, organ damage
and other disease complications in sickle cell patients.
[0057] Clupanodonic acid, also known as omega-3 DPA, is known to be
useful as a platelet aggregation inhibitor and other omega-3 acids
are known to possess anti-inflammatory properties. Vaso-occlusive
events are the source of a number of problematic symptoms in
patients with sickle disease and can occur through platelet
aggregation as well as red blood cell aggregation and white blood
cell aggregation. Some vaso-occlusive events are the result of
thrombosis, which is initiated by thromboxane A2 (TXA2). Omega-3
DPA has been shown to be a potent inhibitor of COX-1 activity (the
enzyme involved in synthesis of TXA2), thus omega-3 DPA can inhibit
the pathways through which vaso-occlusive events occur. The studies
above strongly suggest that omega-3 fatty acids can delay or even
prevent some of the damaging effects of inflammation in sickle cell
patients. Given the lack of effective drugs in treating sickle cell
disease, there is a critical unmet need in developing new therapies
for combatting this debilitating and life-shortening disease.
[0058] Accordingly, any new therapy that reduces the frequency or
severity of inflammation, anemia and/or vaso-occlusive crises in
patients is likely to play a useful role in the treatment of sickle
cell disease. Given the observation that omega-3 fatty acids may be
useful in the treatment of sickle cell disease and improving or
protecting cell health, new formulations, particularly those with a
demonstrable increase in absorption and blood levels of omega-3 and
other fatty acids in the treatment of sickle cell disease are
warranted.
[0059] Macular Degeneration
[0060] Age-related macular degeneration (AMD) is a condition that
results in a loss of vision in the center of the visual field (the
macula). It is a progressive disease and is the leading cause of
severe vision loss in people over age 60. It occurs in "dry" and
"wet" forms and causes damage to the macula, a small spot near the
center of the retina and the part of the eye needed for sharp,
central vision, which lets us see objects that are straight
ahead.
[0061] The macula is the part of the retina that receives most of
the direct light in the eye; is very sensitive to light, with
higher concentrations of rods and cones, and is needed for accurate
central vision. The retina is a network of visual receptors and
nerves that lies on the choroid; a layer of blood vessels that
provides blood to the macula, and the retinal pigment epithelium
(RPE). The RPE is another layer of specialized cells, which
transports the nutrients from the blood vessels and clears waste
products from the macula.
[0062] Dry AMD is an early stage of the disease and may result from
the aging and thinning of macular tissues due to the degeneration
or atrophy that occurs from cell death. It is characterized by the
presence of yellow spots, called drusen, which form in and around
the macula. It is believed these spots are deposits or debris from
deteriorating tissue, which accumulates between the retina and the
choroid due to the inability of the RPE to get rid of the waste and
leads to the eventual deterioration of the photoreceptors.
[0063] Wet AMD refers to the growth of abnormal growth of blood
vessels (angiogenesis) beneath the retina, also referred to as
choroidal neovascularization. These blood vessels leak blood and
fluid into the retina, causing distortion of vision that makes
straight lines look wavy, as well as blind spots and loss of
central vision. These abnormal blood vessels eventually scar,
leading to permanent loss of central vision.
[0064] Dry AMD occurs less often to people who exercise, avoid
smoking and eat healthy food, including vegetables and fish. The
National Eye Institute found in the Age-Related Eye Disease Studies
(AREDS) that daily intake of a combination of vitamin C, vitamin E,
beta-carotene, zinc, and copper can reduce the risk of late AMD by
25 percent and can slow progression of the disease. The AREDS2
trial found that adding lutein and zeaxanthin or omega-three fatty
acids to the original AREDS formulation (with beta-carotene) had no
overall effect on the risk of late AMD. However, the trial also
found that replacing beta-carotene with a 5-to-1 mixture of lutein
and zeaxanthin may help further reduce the risk of late AMD. While
lutein and zeaxanthin appear to be safe regardless of smoking
status of the individual suffering from AMD, high beta-carotene
intake has been linked to an increased risk of lung cancer in
current and former smokers. For wet AMD, angiogenesis inhibitors,
such as for example, BEVACIZUMAB, RANIBIZUMAB, PEGAPTANIB, AND
AFLIBERCEPT can be injected into one's eye to stop new blood
vessels from growing.
[0065] Oxidative damage, resulting from excess production of
reactive oxygen species (ROS) has been implicated in the
progression AMD. Retinal pigmented epithelial (RPE) cells are
highly metabolically active and there is strong evidence that the
RPE cells are extremely sensitive to oxidative stress. It has been
reported that the pathophysiology of AMD is due to cumulative
oxidative damage to RPE cells resulting from an imbalance between
the generation of ROS and the ability of these cells to destroy
and/or protect against ROS damage. Strategies for protecting RPE
cells against oxidative damage may be particularly important in
maintaining retinal function and both preventing and treating
AMD.
[0066] AREDS was designed to evaluate the prevention and treatment
effects of high doses of antioxidants and zinc on individuals with
AMD. AREDS2, was conducted to evaluate the effects on progression
and treatment of AMD patients of a modified version of the AREDS
formulation, including omega-3 fatty acids. The study showed no
AMD-related benefit from the introduction of the omega-3 fatty
acids into the treatment regimen. However, because the AREDS2
formulation did not include formulas with self-micellizing
properties, the AREDS2 study was unable to evaluate an efficacious
bioavailability of the omega-3 fatty acids on AMD. Furthermore, the
AREDS2 study was unable to evaluate predictable, non-food dependent
bioavailability of fat-soluble vitamins because said vitamins were
not co-administered with self-micellizing formulations. Therefore,
the benefits of administering omega-3 fatty acids with
self-micellizing formulations would not have been recognized by
previous studies.
[0067] The lack of any approved effective medications for AMD
warrants the critical need for new therapies to treat AMD.
[0068] Neurodegenerative Diseases
[0069] Neurodegenerative disease is an umbrella term for a range of
conditions, which primarily affect the neurons in the human brain.
Examples of the more common neurodegenerative diseases include
Parkinson's Disease (PD), Alzheimer's Disease (AD), Amyotrophic
Lateral Sclerosis (ALS or Lou Gehrig's Disease), Epilepsy, and
Multiple Sclerosis (MS).
[0070] PD is a neurodegenerative disorder primarily characterized
by motor symptoms but which also includes several other
pathological features such as autonomic system failures, mood
disorders, and cognitive deficits. Conventional treatment may
involve medication that is primarily aimed at increasing dopamine
activity either by providing the precursor (raw material) in the
form of levodopa (L-Dopa), or by stimulating dopamine receptors
(essentially mimicking dopamine) through the use of a dopamine
agonist drug. Also used are drugs called COMT inhibitors which can
help the levodopa to be more effective and MAO-B inhibitors which
prevent dopamine from breaking down so the limited supply is longer
lasting. Other therapies may include physiotherapy, osteopathy,
remedial movement, massage, speech therapy, psychological therapy
and in some cases surgery (deep brain stimulation). Current
pharmacological options for the disease are limited to symptom
management and their long-term use leads to important side
effects.
[0071] MS is a nervous system disease that affects your brain and
spinal cord. It damages the myelin sheath, the material that
surrounds and protects your nerve cells. This damage slows down or
blocks messages between your brain and your body, leading to the
symptoms of MS. They can include visual disturbances, muscle
weakness, trouble with coordination and balance, sensations such as
numbness, prickling, or "pins and needles", and thinking and memory
problems. Although there is no known cure for multiple sclerosis,
several therapies have proven helpful. The primary aims of therapy
are returning function after an attack, preventing new attacks, and
preventing disability. As with any medical treatment, medications
used in the management of MS have several adverse effects.
Available management and/or treatment of the disease is dependent
on the stage of the disease. For example, in relapsing remitting
multiple sclerosis (RRMS), eight disease-modifying treatments have
been approved by regulatory agencies for RRMS including: interferon
beta-la, interferon beta-1b, glatiramer acetate, mitoxantrone,
natalizumab, fingolimod, teriflunomide, and dimethyl fumarate.
There is no treatment available, however, for primary progressive
MS (PPMS).
[0072] ALS, is a rapidly progressive, neuromuscular disease. It
attacks the motor neurons that transmit electrical impulses from
the brain to the voluntary muscles in the body. When they fail to
receive messages, the muscles lose strength, atrophy and die.
RILUZOLE, which extends survival of ALS patients by several months,
is currently the only treatment approved for improving survival,
but does not reverse the damage already done to motor neurons.
BACLOFEN and DIAZEPAM are often prescribed to control the
spasticity caused by ALS, and trihexyphenidyl or amitriptyline may
be prescribed when ALS patients begin having trouble swallowing
their saliva.
[0073] AD is an irreversible, progressive brain disease that slowly
destroys memory and thinking skills, and eventually even the
ability to carry out the simplest tasks. AD is the most common
cause of dementia among older people. Dementia is the loss of
cognitive functioning--thinking, remembering, and reasoning--and
behavioral abilities, to such an extent that it interferes with a
person's daily life and activities. Dementia ranges in severity
from the mildest stage, when it is just beginning to affect a
person's functioning, to the most severe stage, when the person
must depend completely on others for basic activities of daily
living. Four medications are approved by the U.S. Food and Drug
Administration to treat Alzheimer's. DONEPEZIL (ARICEPT.RTM.),
RIVASTIGMINE (EXELON.RTM.), AND GALANTAMINE (RAZADYNE.RTM.) are
used to treat mild to moderate Alzheimer's (donepezil can be used
for severe Alzheimer's as well). MEMANTINE (NAMENDA.RTM.) is used
to treat moderate to severe Alzheimer's. These drugs work by
regulating neurotransmitters (the chemicals that transmit messages
between neurons). They may help maintain thinking, memory, and
speaking skills, and help with certain behavioral problems.
However, these drugs don't change the underlying disease process,
are effective for some but not all people, and may help only for a
limited time.
[0074] Epilepsy is a group of neurological diseases characterized
by epileptic seizures. Epileptic seizures are episodes that can
vary from brief and nearly undetectable to long periods of vigorous
shaking. In epilepsy, seizures tend to recur, and have no immediate
underlying cause while seizures that occur due to a specific cause
are not deemed to represent epilepsy. The cause of most cases of
epilepsy is unknown, although some people develop epilepsy as the
result of brain injury, stroke, brain tumor, and substance use
disorders. Genetic mutations are linked to a small proportion of
the disease. Epileptic seizures are believed to be the result of
excessive and abnormal cortical nerve cell activity in the brain.
Seizures are controllable with medication in about 70% of cases. In
those whose seizures do not respond to medication, then surgery,
neurostimulation, or dietary changes may be considered. The
mainstay treatment of epilepsy is anticonvulsant medications, a
number of which are available. Phenobarbital, phenytoin,
carbamazepine and valproate appear to be equally effective in both
partial and generalized seizures.
[0075] Current epidemiological, preclinical and clinical data
suggest that omega-3 polyunsaturated fatty acids (n-3 PUFAs) may
constitute therapeutic strategy for several disorders of the
central nervous system, including PD, AD, ALS, Epilepsy, Bi-polar
Syndrome, traumatic brain injury, peripheral neuropathy, and MS.
These fatty acids, found most commonly in certain fish and some
plants, are known to help reduce inflammation and oxidative stress
on cells. Both of these processes are known to damage nerve
tissue.
[0076] For example, the concentration of omega-3 fatty acids in
nerve cell membranes has been shown to decrease with age, oxidant
stress, and in neurodegenerative disorders such as Parkinson's
disease. Researchers in Norway have presented evidence of a
systematic omega-3 fatty acid deficit in PD and AD suggesting a
fundamental neurological role for these vital fat molecules.
Supplementation with the omega-3 DHA can favorably modify brain
functions and has been proposed as a nutraceutical in PD and AD.
More recently, the organization Overcoming Multiple Sclerosis (OMS)
conducted a clinical trial looking for a link between omega-3 fatty
acids and MS via the international HOLISM study. The findings
strongly supported a link between consumption of fish and omega-3
supplements and better health for people with MS. Specifically,
people with MS taking flaxseed oil regularly had over 60% fewer
relapses than those who didn't in this large international sample.
Association of fish consumption and omega-3 supplementation with
quality of life, disability and disease activity has been studied
in an international cohort of people with multiple sclerosis.
[0077] Similarly, recent studies show consumption of foods high in
omega-3 fatty acids may help prevent or delay the onset of ALS.
[0078] The studies above strongly suggest that omega-3 fatty acids
can delay or even prevent some neurodegenerative diseases. Given
the lack of effective drugs in treating these diseases, there is a
critical unmet need in developing new therapies for combatting at
least certain neurodegenerative diseases.
[0079] Nutritional Deficiencies and their Effect on Disease State
Manifestations
[0080] Studies have demonstrated that vitamin D may play a
significant role in eye health, specifically in the prevention of
AMD. A high correlation is known to exist between deficient vitamin
D levels and AMD. Vitamin D is believed to act through the
vitamin's anti-inflammatory or anti-angiogenic properties. For this
reason, vitamin D may be useful in the treatment of multiple
inflammation related diseases, including AMD, sickle cell disease,
Alzheimer's, and other diseases.
[0081] Zinc is one of the most abundant trace metals in the human
body. Over 300 enzymes are known to utilize zinc in normal
mammalian function. Zinc deficiency affects many functions that are
directly or indirectly related to declining cognitive performance
in aging individuals, including the progression of Alzheimer's.
Because zinc use is so prolific throughout the body, zinc
deficiency can be causally related to Alzheimer's disease through
multiple pathways. For instance, zinc deficiency can induce insulin
resistance, a risk factor for Alzheimer's. Furthermore, zinc is
believed to be very important in the transport of lipids across the
intestinal lining and therefore affect many diseases related to
lipid deficiencies, including Alzheimer's.
[0082] Alzheimer's disease represents a major health problem in the
US, estimated to be the third leading cause of death, yet the
causes of Alzheimer's disease remain largely unknown and
misunderstood. However, recent research is beginning to demonstrate
a clear link between the progression of the disease and certain
changes in lipids, including fatty acids and fat soluble vitamins,
as well as magnesium and zinc. Markers correlated with progression
of Alzheimer's include changing fatty acid profiles of DHA, EPA,
and arachidonic acid, as well as metabolic syndrome, chronic
inflammation, hypovitaminosis D, and zinc deficiency. Furthermore,
these markers themselves may be correlated.
[0083] Treatment of these correlated conditions can be made through
the introduction of lipids in the correct ratios or amounts.
However, because the absorption of lipids by the digestive system
requires a co-consumption of the dietary fats necessary to form
naturally occurring micelles, the ability to properly dose patients
is often a significant challenge. Further complicating matters, the
over-introduction of certain lipids, such as the fat-soluble
vitamins, can be harmful. Therefore, a need exists to provide the
necessary lipids in a manner which is diet-independent such that
dosing can be more tightly controlled.
[0084] Magnesium contributes to more than three hundred chemical
reactions in mammals, including every ATP-dependent reaction.
Magnesium deficiency is most commonly associated with malabsorption
diseases, including Crohn's disease, gluten-sensitive enteropathy
(celiac disease), and regional enteritis. Resection or bypass of
the small intestine, such as in short bowel disease, typically
leads to malabsorption and magnesium loss. The interaction of
magnesium and vitamin D is particularly important across a plethora
of pathways and body systems. For example, magnesium is required
for both steps in the activation of vitamin D to calcitriol, the
form of the vitamin necessary for calcium absorption. Magnesium is
also required for calcitriol's role in calcium absorption. Mammals
with low magnesium are deficient in both vitamin D and calcitriol,
but the introduction of calcitriol alone does not improve calcium
absorption. Magnesium deficits are correlated to type 2 diabetes
and insulin resistance as a result of higher excretion of magnesium
that results from increased glucose concentrations in the kidneys.
Thus, a need exists to co-administer magnesium with lipids in the
treatment or prevention of certain disease states.
[0085] Vitamins A, D, and K cooperate synergistically not only with
each other, but also with essential minerals like magnesium and
zinc, with dietary fats. Vitamins A, D, and K2 interact
synergistically to support immune health, support bone and teeth
strengthening, and protect soft tissues from calcification. The
interaction of these fat-soluble vitamins with magnesium and zinc
is important. Zinc supports the formation of vesicles involved in
transporting lipids, including the fat-soluble vitamins, across the
intestinal wall.
SUMMARY OF THE INVENTION
[0086] Provided herein, in certain embodiments, are
self-micellizing compositions comprising at least one omega-3 fatty
acid composition (either in the triglyceride, ester, or free fatty
acid form) and at least one surface active agent composition. These
compositions, when combined in specific ratios, are characterized
by their ability to spontaneously form a plurality of stable
micelles within a range of about 1 .mu.m to about 10 .mu.m upon
contact with an aqueous medium.
[0087] In order to determine sample size in a uniform and
reproducible manner, all samples were characterized utilizing the
LS.TM. 13 320 Aqueous Liquid Module (ALM).
[0088] The LS.TM. 13 320 Laser Diffraction Particle Size Analyzer
uses laser diffraction and a patented polarization intensity
differential scattering (PIDS) technology to rapidly determine the
particle size distribution of materials with an overall sizing
range of 0.04 .mu.m to 2000 .mu.m in a single scan with no
extrapolation. This is accomplished with high resolution and
excellent reproducibility.
[0089] In accordance with a standardized protocol, the sample was
heated in its original container at 37.degree. C. for one hour.
After mixing well, approximately 0.25 g of sample was diluted with
5 mL of deionized water. The dilution was manually shaken to form a
homogenous mixture. The preparation was allowed to equilibrate to
room temperature and was subsequently dispensed to the ALM sample
vessel dropwise and analyzed for the particle size distribution
(PSD), which was determined using the Fraunhofer optical model. The
instrument analyzed the PSD from 0.040 .mu.m to 2,000 .mu.m.
[0090] In certain embodiments, the at least one omega-3 fatty acid
(either in the triglyceride, ester, or free fatty acid form) is
selected from the group consisting of hexadecatrienoic acid,
.alpha.-linolenic acid, stearidonic acid, eicosatrienoic acid,
eicosapentaenoic acid, heneicosapentaenoic acid, the omega-3 isomer
of docosapentaenoic acid, docosahexaenoic acid, tetracosapentaenoic
acid, and tetracosahexaenoic acid. In certain embodiments,
self-micellizing compositions may further contain an omega-6 fatty
acid (either in the triglyceride, ester, or free fatty acid form),
for example arachidonic acid (ARA), linoleic acid (LA),
Gamma-linolenic acid (GLA), and omega-6 docosapentaenoic acid
(omega-6 DPA).
[0091] Certain embodiments provide for at least one surface active
agent, a first omega-3 fatty acid (either in the triglyceride,
ester, or free fatty acid form) and a second omega-3 fatty acid
(either in the triglyceride, ester, or free fatty acid form),
wherein the first and second omega-3 fatty acids are different and
are selected from the group consisting of hexadecatrienoic acid,
.alpha.-linolenic acid, stearidonic acid, eicosatrienoic acid,
eicosapentaenoic acid, heneicosapentaenoic acid, the omega-3 isomer
of docosapentaenoic acid, docosahexaenoic acid, tetracosapentaenoic
acid, and tetracosahexaenoic acid.
[0092] Certain embodiments provide for compositions comprising at
least one surface active agent and a first omega-3 fatty acid
(either in the triglyceride, ester or free fatty acid ester form)
and a second omega-3 fatty acid (in either the ester, triglyceride
or free fatty acid form), wherein the first and second omega-3
fatty acids are different and wherein the ratio of the amount of
the first omega-3 fatty acid to the second omega-3 fatty acid is
1:X where 0<X<1. In certain other embodiments, X can be, for
example, 0.001, 0.0015, 0.002, 0.0025, 0.0055, 0.006, 0.0065,
0.007, 0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02,
0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07,
0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.55, 0.6,
0.65, 0.7, 0.75, 0.8, 0.85, 0.9, or, 0.95.
[0093] In certain embodiments, the compositions comprise at least
one surface active agent and a first and second omega-3 fatty acid,
wherein the first omega-3 fatty acid (in either the triglyceride,
ester, or free fatty acid form) is docosahexaenoic acid (DHA) and
the second omega-3 fatty acid (in either the triglyceride, ester,
or free fatty acid form) is selected from the group consisting of
hexadecatrienoic acid, .alpha.-linolenic acid, stearidonic acid,
eicosatrienoic acid, eicosapentaenoic acid, heneicosapentaenoic
acid, the omega-3 isomer of docosapentaenoic acid,
tetracosapentaenoic acid, and tetracosahexaenoic acid and
combinations thereof.
[0094] In certain other embodiments, the compositions comprise at
least one surface active agent and a first and second omega-3 fatty
acid, wherein the first omega-3 fatty acid (in either the
triglyceride, ester, or free fatty acid form) is eicosapentaenoic
acid (EPA) and the second omega-3 fatty acid (in either the
triglyceride, ester, or free fatty acid form) is selected from the
group consisting of hexadecatrienoic acid, .alpha.-linolenic acid,
stearidonic acid, eicosatrienoic acid, heneicosapentaenoic acid,
docosahexaenoic acid (DHA), the omega-3 isomer of docosapentaenoic
acid (DPA), tetracosapentaenoic acid, and tetracosahexaenoic acid
and combinations thereof.
[0095] In certain embodiments, the compositions comprise at least
one surface active agent and a first and second omega-3 fatty acid,
wherein the first omega-3 fatty acid (in either the triglyceride,
ester, or free fatty acid form) is DHA and the second omega-3 fatty
acid (in either the triglyceride, ester, or free fatty acid form)
is EPA, wherein the ratio of the amount of DHA:EPA is 1:X, where
0<X<1. In certain other embodiments, X can be, for example,
0.001, 0.0015, 0.002, 0.0025, 0.0055, 0.006, 0.0065, 0.007, 0.0075,
0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03,
0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08,
0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.55, 0.6, 0.65, 0.7,
0.75, or 0.8.
[0096] In certain embodiments, the compositions comprise at least
one surface active agent and a first and second omega-3 fatty acid,
wherein the first omega-3 fatty acid (in either the triglyceride,
ester, or free fatty acid form) is EPA and the second omega-3 fatty
acid (in either the triglyceride, ester, or free fatty acid form)
is DHA, wherein the ratio of the amount of EPA:DHA is 1:X, where
0<X<1. In certain other embodiments, X can be, for example,
0.001, 0.0015, 0.002, 0.0025, 0.0055, 0.006, 0.0065, 0.007, 0.0075,
0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03,
0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08,
0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.55, 0.6, 0.65, 0.7,
0.75, 0.8, 0.85, 0.9, or 0.95.
[0097] In certain embodiments, the compositions described herein
optionally comprise a third fatty acid (either in the triglyceride,
ester, or free fatty acid form), wherein the third fatty acid is
different from the first and second omega-3 fatty acids. In such
compositions, the ratio of the amount of the first omega-3 fatty
acid to the second and third omega-3 fatty acids is 1:X, where X is
the combined amount of the second and third fatty acids and where
0<X<1. For example, if the first omega-3 fatty acid is DHA
and the second omega-3 fatty acid is EPA, the third omega-3 fatty
acid may be selected from the group consisting of hexadecatrienoic
acid, .alpha.-linolenic acid, stearidonic acid, eicosatrienoic
acid, heneicosapentaenoic acid, the omega-3 isomer of
docosapentaenoic acid, tetracosapentaenoic acid, and
tetracosahexaenoic acid. The ratio of the amount of DHA:(EPA+third
omega-3 fatty acid) is 1:X, where X is the amount of EPA plus the
amount of the third omega-3 fatty acid, and where 0<X<1. In
certain embodiments, X can be, for example, 0.001, 0.0015, 0.002,
0.0025, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009,
0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05,
0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1,
0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7,
0.75, 0.8, 0.85, 0.9, or 0.95. In certain embodiments, the third
fatty acid can be an omega-6 fatty acid such as arachidonic acid
(ARA), linoleic acid (LA), gamma-linolenic acid (GLA) or the
omega-6 isomer of DPA (either in the triglyceride, ester, or free
fatty acid form).
[0098] In certain embodiments, the omega-3 fatty acids (either in
the triglyceride, ester, or free fatty acid form) are substantially
pure.
[0099] In certain embodiments, the omega-3 fatty acids are in the
form of triglycerides.
[0100] In certain embodiments, the omega-3 fatty acids are in the
form of esters.
[0101] In certain embodiments, the omega-3 fatty acid ester is the
ethyl ester derivative.
[0102] In certain embodiments, the omega-3 fatty acids are in the
form of free fatty acids.
[0103] In certain embodiments, the compositions comprising the at
least one surface active agent and the omega-3 fatty acids in
either the triglyceride or ester forms are free of free fatty acid
forms.
[0104] In certain embodiments, the compositions are substantially
free of active agents other than the omega-3 fatty acids (either in
the triglyceride, ester, or free fatty acid form).
[0105] In certain embodiments, the omega-3 fatty acid or
combinations of omega-3 fatty acids (in either the triglyceride,
ester, or free fatty acid form) comprises, for example, 60%, 65%,
70%, 75%, 80%, 85%, 90% or 95% (wt/wt) of the composition.
[0106] In certain embodiments, the compositions described herein
comprise DHA at, for example, from 50% to 99% (wt/wt) of the
compositions. For example, in certain embodiments, the DHA is
present at 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%
(wt/wt) of the compositions. In certain other embodiments, the DHA
is present at least, for example, 90% (wt/wt) of the compositions
described herein. In still other embodiments, the DHA is present at
least, for example, 95% (wt/wt) of the compositions described
herein.
[0107] In certain embodiments, the compositions described herein
comprise EPA at, for example, from 0% to 50% (wt/wt) of the
compositions. For example, in certain embodiments, the EPA is
present at <1% (wt/wt) of said compositions. In certain other
embodiments, the EPA can be present at, for example, 2%, 4%, 6%,
8%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 32%,
34%, 36%, 38%, 40%, 42%, 44%, 46%, 48% or 50% (wt/wt) of said
compositions. In still other embodiments, the EPA can be present at
<5% (wt/wt) of said compositions. For example, in certain
embodiments, the EPA can be present at 0%, 0.1%, 0.2%, 0.3%, 0.4%,
0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, or
4.5% (wt/wt) of said compositions.
[0108] In certain embodiments, the compositions described herein
comprise an omega-3 fatty acid other than DHA and EPA at, for
example, from >0% to 10% (wt/wt) of said compositions. For
example, said omega-3 fatty acid other than DHA and EPA can be
present, in certain embodiments at 0.1%, 0.2%, 0.3%, 0.4%, 0.5%,
0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%,
5.5%, %, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10% (wt/wt) of said
compositions. In such embodiments, the omega-3 fatty acid other
than EPA and DHA can be selected from the group consisting of
hexadecatrienoic acid, .alpha.-linolenic acid, stearidonic acid,
eicosatrienoic acid, heneicosapentaenoic acid, the omega-3 isomer
of docosapentaenoic acid, tetracosapentaenoic acid, and
tetracosahexaenoic acid.
[0109] In certain embodiments, the compositions described herein
comprise 90%, 5%, and 5% (wt/wt) of said compositions DHA, ARA, and
EPA respectively.
[0110] In certain embodiments, the compositions described herein
comprise 95%, 5%, and <1% (wt/wt) of said compositions DHA, ARA,
and EPA respectively.
[0111] In certain embodiments, the omega-3 fatty acid compositions
described herein comprise at least one surface active agent
selected from the group consisting of at least one nonionic surface
active agents, cationic surface active agents, anionic surface
active agents, zwitterionic surface active agents, and combinations
thereof.
[0112] In certain embodiments, the surface active agent is selected
from the group consisting of at least one anionic surface active
agent, at least one non-ionic surface active agent, and a
combination thereof.
[0113] In certain embodiments comprising at least one surface
active agent, the at least one surface active agent has a
hydrophilic-lipophilic balance (HLB) of 8.0.
[0114] In certain embodiments comprising at least one surface
active agent, the surface active agent can be a non-ionic surface
active agent selected from the group consisting of at least one
polysorbate, at least one poloxamer, and a combination thereof.
[0115] In certain embodiments, the at least one surface active
agent comprises a polysorbate present from, for example, 15%
(wt/wt) to 35% (wt/wt) of the composition.
[0116] In certain embodiments, the polysorbate is present at, for
example, 31% (wt/wt) of the composition.
[0117] In certain embodiments, the polysorbate is polysorbate
80.
[0118] In certain other embodiments, the at least one surface
active agent comprises a poloxamer present from, for example, 0.1%
(wt/wt) to 5% (wt/wt) of the composition.
[0119] In certain embodiments, the poloxamer is present at, for
example, 0.7% (wt/wt) of the composition.
[0120] In certain embodiments, the poloxamer is Poloxamer 237, also
known as Pluronic.RTM. F87.
[0121] In certain embodiments, the compositions described herein
comprise a combination of polysorbate 80 and the Poloxamer 237
(Pluronic.RTM. F87)
[(HO(C.sub.2H.sub.4O).sub.64(C.sub.3H.sub.6O).sub.37(C.sub.2H.sub.4O).sub-
.64H].
[0122] In certain embodiments, the combination of polysorbate and
poloxamer comprise at least, for example, 25% (wt/wt) of said
composition.
[0123] In certain embodiments, the composition further comprises at
least one antioxidant. In such embodiments the at least one
antioxidant is selected from the group consisting of a tocopherol,
a tocotrienol, and combinations thereof. In such embodiments, the
tocopherol, tocotrienol and combinations thereof is present from,
for example, from about 0.01% to about 5% by weight of the
compositions. In certain such embodiments, the tocopherols,
tocotrienols and combinations thereof can be present at, for
example, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%,
0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, or 6%
(wt/wt) of the compositions. In certain such embodiments, the
tocopherols, tocotrienols, and combinations thereof can be present
at, for example, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%,
or 6% (wt/wt) of the compositions. In certain embodiments further
comprising at least one antioxidant, the antioxidant is d-gamma
tocotrienol present at, for example, 3%, 3.35%, 4%, 4.35%, 5%, or
5.35% (wt/wt) of the composition. In certain embodiments further
comprising at least one antioxidant, the antioxidant is
.alpha.-tocopherol present at, for example, 2% (wt/wt) of the
composition.
[0124] Certain embodiments provide for compositions comprising at
least one terpene. Said present can be present from, for example,
0.1% (wt/wt) to 5% (wt/wt). In certain embodiments, the terpene is
present at, for example, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%,
0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%,
4.5% or 5% (wt/wt) of said composition. The terpene can be
d-limonene.
[0125] In embodiments comprising substantially pure d-limonene, the
d-limonene is from about, for example, 95% to about 98% pure. In
certain embodiments, the substantially pure d-limonene is at least,
for example, 95%, 96%, 97% or 98%, 99% pure.
[0126] Certain embodiments provide for compositions comprising
natural orange-oil.
[0127] In certain embodiments, the bioavailability of the omega-3
fatty acids (in either the triglyceride, ester, or free fatty acid
form) included in the compositions described herein is
substantially the same when administered with or without food,
i.e., substantially independent of food effect.
[0128] While not wishing to be bound to any particular theories, it
has been observed that compositions comprising at least one omega-3
fatty acid (either in the triglyceride, ester or free fatty acid
ester form) and at least one surface active agent, which are
present in an amount and a combination effective to cause said
compositions to self-micellize when in contact with an aqueous
medium, can be characterized by their ability to spontaneously form
a plurality of stable micelles having a particle size within a
range of about 1 .mu.m to about 10 .mu.m, upon contact with the
aqueous medium. The plurality of micelles produced in this manner
demonstrate enhanced bioavailability of said at least one omega-3
fatty acid, while eliminating any food effect, and improving
patient compliance due to the lowered dosage required to reach a
similar bioavailability or AUC (e.g. 1 pill versus 6).
[0129] In at least one embodiment, a method is provided for
treating AMD by administering the compositions described herein to
a human in need of such administration. The human can be an adult,
a child, an adolescent, or and infant, such as for example a
neonate. In certain embodiments, a method is provided for treating
Juvenile Macular Degeneration, which includes Stargardt's disease,
Best disease, and juvenile retinoschisis by administering the
compositions described herein to a human juvenile.
[0130] In certain embodiments, the compositions may comprise at
least one omega-3 fatty acid, and at least one non-omega-3 fatty
acid nutritional supplement agent, wherein the omega-3 fatty acid
(in either the triglyceride, ester, or free fatty acid form) is
docosahexaenoic acid (DHA) (in either the triglyceride, ester, or
free fatty acid form) and wherein the non-omega-3 fatty acid active
agent or non-omega-3 fatty acid nutritional supplement may be
selected from the group consisting of vitamin C, vitamin E,
beta-carotene, magnesium, zinc (including as zinc oxide), and
copper (including as cupric oxide), and combinations thereof, and
at least one surface active agent, in an amount and a combination
effective to cause said compositions to spontaneously
self-micellize when in contact with an aqueous medium, thereby
forming a plurality of stable micelles having a particle size
within a range of about 1 to about 10 .mu.m.
[0131] In certain embodiments, the compositions comprise at least
one surface active agent, at least one omega-3 fatty acid, and a
non-omega-3 fatty acid active agent, wherein the omega-3 fatty acid
(in either the triglyceride, ester, or free fatty acid form) is
eicosapentaenoic acid (EPA) (in either the triglyceride, ester, or
free fatty acid form) and wherein the non-omega-3 fatty acid active
agent is selected from the group consisting of vitamin C, vitamin
E, beta-carotene, magnesium, zinc (including as zinc oxide), and
copper (including as cupric oxide), and combinations thereof.
[0132] In certain embodiments, the compositions comprise at least
one surface active agent, a first omega-3 fatty acid, a second
omega-3 fatty acid, and a non-omega-3 fatty acid active agent,
wherein the first omega-3 fatty acid (in either the triglyceride,
ester, or free fatty acid form) is EPA and the second omega-3 fatty
acid (in either the triglyceride, ester, or free fatty acid form)
is DHA, wherein the ratio of the amount of EPA:DHA is 1:X, where
0<X<1 and wherein the non-omega-3 fatty acid active agent is
selected from the group consisting of vitamin C, vitamin E,
beta-carotene, magnesium, zinc (including as zinc oxide), and
copper (including as cupric oxide), arachidonic acid (ARA),
linoleic acid (LA), gamma-linolenic acid (GLA) or the omega-6
isomer of DPA and combinations thereof.
[0133] Abnormal endothelial cell migration and proliferation can
lead to adherent neovascularization. In the wet form of age-related
macular degeneration (AMD), neovascularization can create severe
vision problems. Current treatments include the use of known
anti-angiogenesis medications. While not wishing to be bound to any
particular theories, it is believed that compositions comprising
omega-3 DPA can prevent angiogenesis through the vascular
endothelial growth factor (VEGF) pathway.
[0134] In at least one embodiment, a method is provided for
treating AMD by administering, to a patient in need thereof, a
therapeutically effective amount of a composition comprising
omega-3 DPA fatty acid (either in the triglyceride, ester or free
fatty acid ester form) and at least one surface active agent, in an
amount and a combination effective to cause said compositions to
spontaneously self-micellize when in contact with an aqueous
medium, thereby forming a plurality of stable micelles having a
particle size within a range of about 1 .mu.m to about 10
.mu.m.
[0135] In at least one embodiment, a method is provided for
treating fat malabsorption syndromes by administering the
compositions described herein to a human in need of such
administration. The human can be an adult, an adolescent, or and
infant, such as for example a neonate.
[0136] In at least one embodiment, a method is provided for
treating fat malabsorption syndrome by administering the
compositions described herein to a human suffering from SBS or
Barrett's Syndrome.
[0137] In at least one embodiment, a method is provided for
treating sickle cell disease by administering the compositions
described herein to a human in need of such administration. The
human can be an adult, a child, an adolescent, or and infant, such
as for example a neonate.
[0138] In at least one embodiment, a method is provided for
treating NAFLD and/or NASH by administering the compositions
described herein to a human in need of such administration. The
human can be an adult, a child, an adolescent, or and infant, such
as for example a neonate.
[0139] In at least one embodiment, a method is provided for
treating neurodegenerative diseases, such as for example,
Parkinson's Disease (PD), Alzheimer's Disease (AD), Multiple
Sclerosis (MS), Epilepsy, and Amyotrophic Lateral Sclerosis (ALS),
by administering the compositions described herein to a human in
need of such administration. The human can be an adult, a child, an
adolescent, or and infant, such as for example a neonate.
[0140] In at least one embodiment of the invention, a method is
provided for preventing or treating Alzheimer's disease by
administering the compositions described herein to a subject in
need of such administration, wherein said compositions may include
at least one fat-soluble vitamin, at least one omega-3 fatty acid
(in triglyceride, ester, or free fatty acid form), one or more
minerals, such as magnesium, manganese, zinc, copper, selenium, and
combinations thereof, and at least one surface active agent in an
amount and a combination effective to cause said compositions to
spontaneously self-micellize when in contact with an aqueous
medium, thereby forming a plurality of stable micelles having a
particle size within a range of about 1 .mu.m to about 10
.mu.m.
[0141] In at least one embodiment of the invention, a method is
provided for preventing or treating age-related macular
degeneration by administering the compositions described herein to
a subject in need of such administration, wherein said compositions
may include at least one fat-soluble vitamin, an omega-3 fatty acid
(in triglyceride, ester, or free fatty acid form, and one or more
minerals, such as magnesium, manganese, zinc, copper, selenium, and
combinations thereof, and at least one surface active agent in an
amount and a combination effective to cause said compositions to
spontaneously self-micellize when in contact with an aqueous
medium, thereby forming a plurality of stable micelles having a
particle size within a range of about 1 .mu.m to about 10
.mu.m.
[0142] In at least one embodiment of the invention, a method is
provided for preventing or treating diseases by administering the
compositions described herein to a subject in need of such
administration, wherein said compositions may include at least one
fat-soluble vitamin, an omega-3 fatty acid (in triglyceride, ester,
or free fatty acid form), and one or more minerals, such as
magnesium, manganese, zinc, copper, selenium, and combinations
thereof, and at least one surface active agent in an amount and a
combination effective to cause said compositions to spontaneously
self-micellize when in contact with an aqueous medium, thereby
forming a plurality of stable micelles having a particle size
within a range of about 1 .mu.m to about 10 .mu.m., and wherein
said diseases may include one or more of metabolic syndrome,
chronic inflammation, hypovitaminosis D, zinc deficiency, Crohn's
disease, diseases related to calcium deficiency, Alzheimer's
disease, age-related macular degeneration, short bowel syndrome,
sickle cell disease, irritable bowel syndrome, clotting disorders,
hypertriglyceridemia, PSC, non-alcoholic fatty liver disease,
epilepsy, age-related dementia, diabetic retinopathy, insulin
resistance, diabetes, muscular dystrophy, microvillus inclusion
disease (also known as Davidson's disease, congenital microvillus
atrophy), intestinal epithelial dysplasia (IED), also known as
tufting enteropathy, syndromic diarrhea (SD), also known as
phenotypic diarrhea (PD) or tricho-hepato-enteric syndrome (THE),
and cystic fibrosis.
[0143] In at least one embodiment of the invention, a method is
provided for treating short bowel syndrome by administering the
compositions described herein to a subject in need of such
administration, wherein said compositions may include at least one
fat-soluble vitamin, an omega-3 fatty acid (in triglyceride, ester,
or free fatty acid form), and one or more minerals, such as
magnesium, manganese, zinc, copper, selenium, and combinations
thereof, and an omega-6 fatty acid (in triglyceride, ester, or free
fatty acid form), for example arachidonic acid (ARA), linoleic acid
(LA), gamma-linolenic acid (GLA) or the omega-6 isomer of DPA, and
at least one surface active agent in an amount and a combination
effective to cause said compositions to spontaneously
self-micellize when in contact with an aqueous medium, thereby
forming a plurality of stable micelles having a particle size
within a range of about 1 .mu.m to about 10 .mu.m.
[0144] In certain embodiments, the compositions described herein
self-micellize in an aqueous medium. In certain other embodiments,
the aqueous medium is water. In certain other embodiments, the
aqueous medium has an acidic pH. In certain other embodiments, the
aqueous medium is 0.1N HCl.
[0145] In certain embodiments, the compositions described herein
self-micellize in an aqueous medium, wherein the micelles have a
diameter from about 1 .mu.m to about 10 .mu.m. In certain
embodiments, the compositions described herein self-micellizes in
an aqueous medium having an acidic pH, wherein the micelles have a
diameter from about 1 .mu.m to about 10 .mu.m. In certain other
embodiments, the compositions described herein self-micellizes in
0.1N HCL, wherein the micelles have a diameter from about 1 .mu.m
to about 10 .mu.m. In certain embodiments, the micelles may have an
average diameter of about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10
.mu.m.
[0146] In certain embodiments, the compositions described herein
minimize or eliminate at least one side effect when compared to the
administration of a composition comprising omega-3 fatty acids
known to one of skill in the art. Non-limiting examples of the side
effects include regurgitation, frequency of burping,
gastroesophageal reflux disease (GERD), bloating, increased
intestinal gas, fish taste, fishy breath, fish smell, nausea,
diarrhea, or combinations thereof.
[0147] In certain embodiments, the compositions described herein
comprise d-limonene or natural orange oil. Such compositions can
minimize or eliminate at least one side effect from the
administration of a composition of the present disclosure when
compared to the administration of a composition comprising omega-3
fatty acids substantially free of d-limonene or natural orange oil.
Non-limiting examples of the side effects include regurgitation,
frequency of burping, gastroesophageal reflux disease (GERD),
bloating, increased intestinal gas, fish taste, fishy breath, fish
smell, nausea, diarrhea, or combinations thereof.
[0148] Certain embodiments of the compositions described herein can
be administered to a human subject in need of such administration
in conjunction with a non-omega-3 fatty acid nutritional supplement
or a non-omega-3 fatty acid active agent for the treatment of
macular degeneration. Non-limiting examples of non-omega-3 fatty
acid nutritional supplements may include a combination of vitamin
C, vitamin E, beta-carotene, zinc, copper, magnesium, manganese,
calcium, vitamin A, vitamin D, vitamin K (includes K1 and/or K2),
lutein and zeaxanthin.
[0149] Certain embodiments of the compositions described herein can
be administered to a human subject in need of such administration
in conjunction with a non-omega-3 fatty acid active agent for the
treatment of SBS. Non-limiting examples of such non-omega-3 fatty
acid active agents can include L-Glutamine (NUTRESTORE.RTM.),
recombinant somatotropin (ZORBTIVE.RTM.), and teduglutide
(GATTEX.RTM.). Other such non-omega-3 fatty acid active agents may
include conjugated bile acids or opium tincture.
[0150] Certain embodiments of the compositions described herein can
be administered to a human subject in need of such administration,
either alone, or in conjunction with a non-omega-3 fatty acid
nutritional supplement or a non-omega-3 fatty acid active agent.
For example, (1) in the treatment of a fat malabsorption syndrome,
e.g. short bowel syndrome, a composition within the purview of the
present invention will include substantially pure DHA, or a
combination of substantially pure DHA and arachidonic acid in
combination with a surface active agent which is a combination
including about 31% of Polysorbate 80 and about 0.7% Poloxamer 237;
(2) in the treatment of hypertriglyceridemia or hypocholesteremia,
a composition within the purview of the present invention will
include a combination of EPA and DHA, or a composition including
EPA and a therapeutically effective amount of one or more statins,
in combination with a surface active agent which is a combination
including about 31% of Polysorbate 80 and about 0.7% Poloxamer 237;
(3) in the treatment of non-alcoholic fatty liver disease (NAFLD)
or non-alcoholic steatohepatitis (NASH), a composition within the
purview of the present invention will include substantially pure
DHA, in combination with a surface active agent which is a
combination including about 31% of Polysorbate 80 and about 0.7%
Poloxamer 237; (4) in the treatment of sickle cell disease (SCD), a
composition within the purview of the present invention will
include substantially pure DHA alone, substantially pure omega-3
DPA alone, or a combination of said DHA and DPA, in combination
with a surface active agent which is a combination including about
31% of Polysorbate 80 and about 0.7% Poloxamer 237, in a
therapeutically effective amount to treat sickle cell disease
symptoms, such as vaso-occlusive events, sequestration crisis,
avascular necrosis, stroke, and sickle cell anemia, among other
known maladies, by decreasing the aggregation of platelets, red
blood cells, and/or white blood cells.
[0151] Certain embodiments of the compositions described herein can
be administered to a human subject in need of such administration
in conjunction with a non-omega-3 fatty acid nutritional supplement
or a non-omega-3 fatty acid active agent for the treatment of
sickle cell disease. Non-limiting examples of non-omega-3 fatty
acid active agents can include pain medications, antibiotics, a
therapeutically effective amount of a non-steroidal
anti-inflammatory medication, illustrated by, albeit not limited to
a particular dosage of acetysalicylic acid (aspirin), hydroxyurea,
and anti-inflammatory medications. Non-limiting examples of
non-omega-3 fatty acid nutritional supplements may include a
combination of folic acid, vitamin C, vitamin E, beta-carotene,
zinc, copper, magnesium, manganese, calcium, vitamin A, vitamin D,
vitamin K (includes K1 and/or K2), lutein and zeaxanthin.
[0152] Certain embodiments of the compositions described herein can
be administered to a human subject in need of such administration
in conjunction with a non-omega-3 fatty acid nutritional supplement
or a non-omega-3 fatty acid active agent for the treatment of NAFLD
and/or NASH. Non-limiting examples of non-omega-3 fatty acid active
agents can include, lipid lowering or cholesterol lowering agents
selected from the group consisting of cholesterol absorption
inhibitors, bile acid sequestrants/resins, statins, niacin and
derivatives, MTP inhibitors, fibrates and CETP inhibitors, insulin
sensitizers, hypolipidemics, anti-inflammatory medications, and
thiazolidinediones.
[0153] Certain embodiments of the compositions described herein can
be administered to a human subject in need of such administration
in conjunction with a non-omega-3 fatty acid nutritional supplement
or a non-omega-3 fatty acid active agent for the treatment of PSC
or Alagille Syndrome. Non-limiting examples of non-omega-3 fatty
acid active agents can, upon approval by a respective regulatory
agency, may include 6-alpha-ethylchenodeoxycholic acid and salts
thereof, and/or
(4R,5R)-1-[[4-[[4-[3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hyd-
roxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]methyl]phenyl]methyl]-4-aza-1-
-azoniabicyclo[2.2.2]octane chloride.
[0154] Certain embodiments of the compositions described herein can
be administered to a human subject in need of such administration
in conjunction with a non-omega-3 fatty acid nutritional supplement
or a non-omega-3 fatty acid active agent for the treatment of a
neurodegenerative disease. Non-limiting examples of non-omega-3
fatty acid active agents can include medications generally approved
by a health regulatory body and prescribed by physicians for a
particular neurodegenerative disease, such as for example, PD, AD,
MS, Epilepsy, and ALS.
[0155] In certain embodiments, compositions including substantially
pure DHA in combination with at least one surface active agent, as
described herein, effective to spontaneously form micelles, or
alternatively, pre-formed micelles produced therefrom, are
effective for the treatment of neurodevelopmental psychiatric
disorders, such as Attention Deficit Disorder (ADD) and Attention
Deficit Hyperactivity Disorder (ADHD).
[0156] In at least one embodiment, the compositions described
herein can be manufactured in an oil-filled inert, non-reactive
vessel, non-limiting examples of which include glass vials and
Teflon.RTM. coated vials; with or without light penetrating
inhibitors, including for example amber colored vials. The
composition in said oil-filled vial can be administered to an adult
human, adolescent, or infant, in particular to a neonate, in need
of such administration, by adding an appropriate amount of saline
or other pharmaceutically acceptable solutions to said vial,
thereby initiating the self-micellization process which results in
the formation of the pre-formed micelles as defined herein. The
homogenous solution, containing the pre-formed micelles, can then
be either administered intravenously or added to infant formula for
oral ingestion When administered orally, the compositions described
herein may be administered in the form of a gel or liquid filled
capsule.
[0157] Also provided are kits comprising compositions described
herein as one or more unit dosage forms together with instructions
on using the dosage forms. In certain embodiments, the dosage forms
described herein can be packaged as unit doses, such as for
example, glass crimp or snap top vials with instructions for using
the unit dosage forms. For example, the instructions can be
provided as a package insert or directly on a label attached to the
vial, which can be made of clear or amber glass. The instructions
can include, for example, dosing frequency recommendations,
administration of the dosage forms with or without food, and the
active ingredients comprising the dosage forms.
[0158] In certain embodiments kits are provided, wherein certain
dosage forms comprising the compositions described herein can be
packaged together with other non-omega-3 fatty acid nutritional
supplements or active agents for the treatment of AMD. The kit(s)
comprise one or more unit dosage forms of certain embodiments of
the compositions described herein together with one or more unit
dosage forms comprising the non-omega-3 fatty acid nutritional
supplement or active agent for the treatment of AMD together with
instructions on using the dosage forms. Non-limiting examples of
non-omega-3 fatty acid nutritional supplements include, for
example, a combination of vitamin C, vitamin E, beta-carotene,
zinc, copper, magnesium, manganese, calcium, vitamin A, vitamin D,
vitamins K1 and/or K2, lutein and zeaxanthin. Non-limiting examples
of non-omega-3 fatty acid active agents include, for example,
verteporfin; the antioxidant carotenoids crocin and crocetin, as
found in, for example, Saffron (Crocus sativus); and inhibitors of
angiogenesis, such as for example, BEVACIZUMAB (AVASTIN.RTM.),
RANIBIZUMAB (LUCENTIS.RTM.), PEGAPTANIB (MACUGEN.RTM.) AND
AFLIBERCEPT (EYLEA.RTM.).
[0159] Certain embodiments provide for a functional food(s) for
treating AMD comprising the compositions described herein.
[0160] Certain embodiments provide methods of treating AMD by
administering a functional food comprising the compositions
described herein.
[0161] The instructions can include, for example, dosing frequency,
administration of the dosage forms with or without food, the active
ingredients comprising the dosage forms, and the fat malabsorption
syndromes that would benefit from administration of the dosage
forms.
[0162] In certain embodiments kits are provided, wherein certain
dosage forms comprising the compositions described herein can be
packaged together with other non-omega-3 fatty acid nutritional
supplements for the treatment of a fat malabsorption syndrome. The
kit(s) comprise one or more unit dosage forms of certain
embodiments of the compositions described herein together with one
or more unit dosage forms comprising the non-omega-3 fatty acid
nutritional supplements for the treatment of a fat malabsorption
syndrome together with instructions on using the dosage forms.
[0163] Certain embodiments provide for a functional food(s) for
treating a fat malabsorption syndrome comprising the compositions
described herein.
[0164] Certain embodiments provide methods of treating a fat
malabsorption syndrome by administering a functional food
comprising the compositions described herein.
[0165] In certain embodiments kits are provided, wherein certain
dosage forms comprising the compositions described herein can be
packaged together with other non-omega-3 fatty acid nutritional
supplements or active agents for the treatment of sickle cell
disease. The kit(s) comprise one or more unit dosage forms of
certain embodiments of the compositions described herein together
with one or more unit dosage forms comprising the non-omega-3 fatty
acid nutritional supplement or active agent for the treatment of
sickle cell disease together with instructions on using the dosage
forms. Non-limiting examples of non-omega-3 fatty acid active
agents and nutritional supplements that can be provided in the kits
include pain medications, antibiotics, folic acid, hydroxyurea, and
anti-inflammatory medications
[0166] Certain embodiments provide for a functional food(s) for
treating sickle cell disease comprising the compositions described
herein.
[0167] Certain embodiments provide methods of treating sickle cell
disease by administering a functional food comprising the
compositions described herein.
[0168] In certain embodiments kits are provided, wherein certain
dosage forms comprising the compositions described herein can be
packaged together with other non-omega-3 fatty acid nutritional
supplements or active agents for the treatment of NAFLD and/or
NASH. The kit(s) comprise one or more unit dosage forms of certain
embodiments of the compositions described herein together with one
or more unit dosage forms comprising the non-omega-3 fatty acid
nutritional supplement or active agent for the treatment of NAFLD
and/or NASH together with instructions on using the dosage forms.
Non-limiting examples of non-omega-3 fatty acid active agents that
can be provided in the kits include, lipid lowering or cholesterol
lowering agents selected from the group consisting of cholesterol
absorption inhibitors, bile acid sequestrants/resins, statins,
niacin and derivatives, MTP inhibitors, fibrates and CETP
inhibitors, insulin sensitizers, hypolipidemics, anti-inflammatory
medications, and thiazolidinediones.
[0169] Certain embodiments provide for a functional food(s) for
treating NAFLD and/or NASH comprising the compositions described
herein. Certain embodiments provide methods of treating NAFLD
and/or NASH by administering a functional food comprising the
compositions described herein.
[0170] In certain embodiments kits are provided, wherein certain
dosage forms comprising the compositions described herein can be
packaged together with other non-omega-3 fatty acid nutritional
supplements or active agents for the treatment of PSC. The kit(s)
comprise one or more unit dosage forms of certain embodiments of
the compositions described herein together with one or more unit
dosage forms comprising the non-omega-3 fatty acid nutritional
supplement or active agent for the treatment of PSC together with
instructions on using the dosage forms. Non-limiting examples of
non-omega-3 fatty acid active agents that can, upon approval by a
respective regulatory agency, be provided in the kits include
6-alpha-ethylchenodeoxycholic and/or
(4R,5R)-1-[[4-[[4-[3,3-Dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hyd-
roxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]methyl]phenyl]methyl]-4-aza-1-
-azoniabicyclo[2.2.2]octane Chloride.
[0171] Certain embodiments provide for a functional food(s) for
treating PSC comprising the compositions described herein.
[0172] Certain embodiments provide methods of treating PSC by
administering a functional food comprising the compositions
described herein.
[0173] In certain embodiments kits are provided, wherein certain
dosage forms comprising the compositions described herein can be
packaged together with other non-omega-3 fatty acid nutritional
supplements or active agents for the treatment of neurodegenerative
disease. The kit(s) comprise one or more unit dosage forms of
certain embodiments of the compositions described herein together
with one or more unit dosage forms comprising the non-omega-3 fatty
acid nutritional supplement or active agent for the treatment of a
neurodegenerative disease together with instructions on using the
dosage forms. Non-limiting examples of non-omega-3 fatty acid
active agents that can be provided in the kits include medications
generally approved by a health regulatory body and prescribed by
physicians for a particular neurodegenerative disease.
[0174] Certain embodiments provide for a functional food(s) for
treating neurodegenerative diseases, such as for example, PD, AD,
MS, Epilepsy, or ALS, comprising the compositions described
herein.
[0175] Certain embodiments provide methods of treating
neurodegenerative diseases, such as for example, PD, AD, MS,
Epilepsy, or ALS, by administering a functional food comprising the
compositions described herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0176] FIGS. 1A-1D illustrate plasma fatty acid profiles (mol %) in
the SBS treatment group (dark gray) versus Control (light gray)
groups. DHA: Docosahexaenoic acid; EPA: Eicosapentaenoic acid; AA:
Arachidonic acid, LA, Linoleic acid. Sample time 0=day of surgery;
1=postoperative day (POD) 1/AM; 2=POD 1/PM; 3=POD 2/AM; 4=POD 2/PM;
5=POD 3/AM; 6=POD 3/PM; 7=POD 4/AM.
[0177] FIGS. 2A-2D illustrate intestinal morphology before and
after surgical induction of SBS and DHA treatment. SBS: Short bowel
syndrome; DHA: Docosahexaenoic acid; Black bars represent tissue
morphometry from samples obtained at the time of surgery (Day 0);
Striped bars represent tissue morphometry from samples obtained at
the end of the study protocol (Final).
[0178] FIG. 3 shows the mean body weight on the day of sacrifice of
the mice between each of the Control, Second Treatment Group and
Third Treatment Group.
[0179] FIG. 4 shows .alpha.-SMA mRNA gene expression in mice in
each of the control, NAFLD and NASH groups treated with either
Vehicle or Composition 1 at the end of the treatment period.
[0180] FIG. 5 shows plasma alanine transaminase (ALT) levels in
mice between each of the control, NAFLD and NASH group treated with
either Vehicle or Composition at the end of the treatment
period.
[0181] FIG. 6 shows the mean liver weight on the day of sacrifice
of the mice between each of the treatment groups and the
control.
[0182] FIG. 7 shows the mean liver to body weight ratio on the day
of sacrifice of the mice between each of the treatment groups and
the control.
[0183] FIG. 8 shows whole blood glucose in mice between each of the
control, NAFLD and NASH group treated with either Vehicle or
Composition at the end of the treatment period.
[0184] FIG. 9 shows liver triglyceride levels in mice between each
of the control, NAFLD and NASH.
[0185] FIGS. 10 A and 10B show the mean individual EPA (A) and DHA
(B) total lipid concentration-time profiles (baseline-adjusted
change) after a single dose of SC401 during fed and fasting
conditions.
[0186] FIGS. 11A and 11B show the mean individual EPA (A) and DHA
(B) free fatty acid concentration-time profiles (baseline-adjusted
change) after a single dose of SC401 during fed and fasting
conditions.
[0187] FIG. 12 shows mean EPA and DHA total lipid plasma
concentration profiles (.mu.g/ml) (baseline-adjusted) after
administration of a single dose (dose adjusted) of SC401 and
Lovaza.RTM. in fasted conditions.
DETAILED DESCRIPTION
[0188] While the present invention is capable of being embodied in
various forms, the description below of several embodiments is made
with the understanding that the present disclosure is to be
considered as an exemplification of the invention, and is not
intended to limit the invention to the specific embodiments
illustrated. Headings are provided for convenience only and are not
to be construed to limit the invention in any manner. Embodiments
illustrated under any heading may be combined with embodiments
illustrated under any other heading.
[0189] The various embodiments of the invention described herein
may suitably comprise, consist essentially of, or consist of, at
least one surface active agent, at least one omega-3 fatty acid (in
either the triglyceride, ester, or free fatty acid form).
[0190] Certain embodiments of the invention may suitably comprise,
consist essentially of, or consist of at least one surface active
agent and a first and second omega-3 fatty acid, wherein the first
omega-3 fatty acid (in either the triglyceride, ester, or free
fatty acid form) is selected from the group consisting of
hexadecatrienoic acid, .alpha.-linolenic acid, stearidonic acid,
eicosatrienoic acid, eicosapentaenoic acid, heneicosapentaenoic
acid, docosapentaenoic acid, docosahexaenoic acid,
tetracosapentaenoic acid, tetracosahexaenoic acid, and combinations
thereof and the second omega-3 fatty acid (in either the
triglyceride, ester, or free fatty acid form) is selected from the
group consisting of hexadecatrienoic acid, .alpha.-linolenic acid,
stearidonic acid, eicosatrienoic acid, eicosapentaenoic acid,
heneicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic
acid, tetracosapentaenoic acid, tetracosahexaenoic acid, and
combinations thereof, wherein the first and second omega-3 fatty
acids (in either the triglyceride, ester, or free fatty acid form)
are different and wherein the ratio of the first omega-3 fatty acid
(in either the triglyceride, ester, or free fatty acid form):the
second omega-3 fatty acid (in either the triglyceride, ester, or
free fatty acid form) is 1:X, where 0<X<0.3 or
0.5<X<1.
[0191] Certain embodiments of the invention may suitably comprise,
consist essentially of, or consist of at least one surface active
agent and a first and second omega-3 fatty acid, wherein the first
omega-3 fatty acid (in either the triglyceride, ester, or free
fatty acid form) is DHA and the second omega-3 fatty acid (in
either the triglyceride, ester, or free fatty acid form) is
selected from the group consisting of hexadecatrienoic acid,
.alpha.-linolenic acid, stearidonic acid, eicosatrienoic acid,
eicosapentaenoic acid, heneicosapentaenoic acid, docosapentaenoic
acid, tetracosapentaenoic acid, tetracosahexaenoic acid, and
combinations thereof, wherein the ratio of the amount of DHA to the
second omega-3 fatty acid is 1:X, where 0<X<0.3 or
0.5<X<1.
[0192] Certain embodiments of the invention may suitably comprise,
consist essentially of, or consist of at least one surface active
agent and a first and second omega-3 fatty acid, wherein the first
omega-3 fatty acid (in either the triglyceride, ester, or free
fatty acid form) is EPA and the second omega-3 fatty acid (in
either the triglyceride, ester, or free fatty acid form) is
selected from the group consisting of hexadecatrienoic acid,
.alpha.-linolenic acid, stearidonic acid, eicosatrienoic acid,
heneicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic
acid, tetracosapentaenoic acid, tetracosahexaenoic acid, and
combinations thereof, wherein the ratio of the amount of EPA to the
second omega-3 fatty acid is 1:X, where 0<X<0.3 or
0.5<X<1.
Definitions
[0193] As used herein, the term "composition(s)" or
"formulation(s)" includes therapeutic and dietary compositions
including, but not limited to, a dietary supplement, nutraceutical
formulation, or pharmaceutical formulation. Further, the terms
composition, dietary supplement, nutraceutical formulation, and
pharmaceutical formulation are used interchangeably herein.
[0194] As used herein, the term "EPA" refers inclusively to
(5Z,8Z,11Z,14Z,17Z)-eicosa-5,8,11,14,17-pentaenoic acid or
derivatives thereof, including alkyl esters, such as, for example,
the ethyl ester.
[0195] As used herein, the term "DHA" inclusively refers to
(4Z,7Z,10Z,13Z,16Z,19Z)-docosa-4,7,10,13,16,19-hexaenoic acid or
derivatives thereof, including alkyl esters, such as, for example,
the ethyl ester.
[0196] As used herein, the term "DPA" refers inclusively to
docosapentaenoic acid (DPA), wherein "omega-3 DPA" refers to the
omega-3 isomer all-cis-7,10,13,16,19-docosapentaenoic acid
(clupanodonic acid) and "omega-6 DPA" refers to the omega-6 isomer
all-cis-4,7,10,13,16-docosapentaenoic acid (osbond acid).
[0197] As used herein, the term "ARA" refers inclusively to
(5Z,8Z,11Z,14-Z)-Icosa-5,8,11,14-tetraenoic acid or derivatives
thereof, including alkyl esters, such as, for example, the ethyl
ester.
[0198] As used herein, the term linoleic acid "LA", refers
inclusively to (9Z,12Z)-9,12-Octadecadienoic acid or derivatives
thereof, including alkyl esters, such as, for example, the ethyl
ester.
[0199] As used herein, the term "GLA" refers inclusively to
all-cis-6,9,12-octadecatrienoic acid or derivatives thereof,
including alkyl esters, such as, for example, the ethyl ester.
[0200] As used herein, the term "micelle" (plural micelles,
micella, plurality of micelles, or micellae) refers to an aggregate
of molecules, that have assembled into a substantially spherical
core/shell architecture, and are suspended in an aqueous phase. A
typical micelle in aqueous solution forms an aggregate with the
hydrophilic "head" regions in contact with surrounding solvent
and/or in contact with the polar region of one or more surface
active agent(s), sequestering the hydrophobic regions in the
micelle center. Micelles are approximately spherical in shape.
[0201] The term "self-micellizes" or "self-micellization" as used
herein refers to the process in which micelles are formed in an
aqueous medium without the introduction of energy, including
agitation or shearing.
[0202] As used herein, the term "aqueous medium" refers to any
solution or suspension, that comprises in part or in whole water,
including for example, without limitation, water by itself;
phosphate buffered saline at about pH 7.4, soft-drinks, illustrated
by, albeit not limited to Sprite.RTM., apple juice, G-2.RTM. fruit
punch, infant formula or any equivalent mammalian mother's milk
substitutes or analogues, intravenous fluid, and milk and milk
products, including chocolate milk. In certain embodiments, an
aqueous medium comprises at least one fluid having at least one
fatty acid, carbohydrate, lipid, sugar, or combinations thereof. In
certain embodiments, an aqueous medium comprises at least one fluid
having an acidic pH. In certain other embodiments, an aqueous
medium comprises a biological fluid such as, for example and
without limitation, stomach acid. In other embodiments, the aqueous
medium comprises simulated stomach acid comprising 0.1N HCl.
[0203] As used herein, the term "free fatty acid" refers to one or
more polyunsaturated fatty acids that have not been modified or do
not have any other groups attached.
[0204] As used herein, the term "ester" refers to the replacement
of the hydrogen in the carboxylic acid group of a polyunsaturated
fatty acid molecule with another substituent. Typical esters are
known to those in the art, a discussion of which is provided by
Higuchi, T. et al., Pro-drugs as Novel Delivery Systems, Vol. 14,
A.C.S. Symposium Series, Bioreversible Carriers in Drug Design, Ed.
Edward B. Roche, Amer. Pharma. Assoc., Pergamon Press (1987), and
Protective Groups in Organic Chemistry, McOmie ed., Plenum Press,
New York (1973), each of which is incorporated herein by reference
in their entirety. Examples of common esters include methyl, ethyl,
trichloroethyl, propyl, butyl, pentyl, tert-butyl, benzyl,
nitrobenzyl, methoxybenzyl, benzhydryl, monoglyceride, diglyceride,
triglyceride.
[0205] As used herein, the term "monoglyceride" refers to a fatty
acid chain, such as DHA or EPA molecule, covalently bonded to a
glycerol molecule through an ester linkage. As used herein, the
term "diglyceride" refers to a fatty acid chain such as DHA or EPA,
covalently bonded to a glycerol molecule through an ester linkage,
wherein the glycerol molecule is further bonded to one additional
fatty acid chain, which may or may not be DHA or EPA, through one
additional ester linkage. As used herein, the term "triglyceride"
refers to a fatty acid chain, such as DHA or EPA, covalently bonded
to a glycerol molecule through an ester linkage, wherein the
glycerol molecule is further bonded to two additional fatty acid
chains, either or both of which may or may not be DHA or EPA,
through two additional ester linkages.
[0206] As used herein, the term "terpene" refers to the large and
diverse class of organic compounds produced by a variety of plants,
particularly conifers. When terpenes are modified chemically, such
as by oxidation or rearrangement of the carbon skeleton, the
resulting compounds are generally referred to as "terpenoids"
(e.g., carvone). Terpenes and terpenoids are the primary
constituents of the essential oils of many types of plants and
flowers.
[0207] As used herein, the terms "tocopherol", and "tocotrienol",
and "vitamin E" refer to a set of fat-soluble vitamins with
antioxidant properties. For example, the term "tocotrienol" refers
to the family of tocotrienols "alpha-tocotrienol",
"beta-tocotrienol", "gamma-tocotrienol" and "delta-tocotrienol".
Similarly, the term "tocopherol" refers to the family of
tocopherols "alpha-tocopherol", "beta-tocopherol",
"gamma-tocopherol" and "delta-tocopherol".
[0208] As used herein, the term "antioxidant" refers to a molecule
capable of inhibiting the oxidation of other molecules. Oxidation
is a chemical reaction that transfers electrons or hydrogen from a
substance to an oxidizing agent. Oxidation reactions can produce
free radicals. In turn, these radicals can start chain reactions.
When the chain reaction occurs in a cell, it can cause damage or
death to the cell. Antioxidants terminate these chain reactions by
removing free radical intermediates, and inhibit other oxidation
reactions. They do this by being oxidized themselves, so
antioxidants are often reducing agents such as thiols, ascorbic
acid, or polyphenols. Exemplary antioxidants include rosemary oil,
ascorbic acid (vitamin C), glutathione, lipoic acid, uric acid,
carotenes, melatonin, ubiquinol (coenzyme Q), .alpha.-tocopherol
(vitamin E), acorbyl palmitate, butylated hydroxyanisole, butylated
hydroxytoluene, monothioglycerol, and potassium metabisulfite.
[0209] As used herein, a "pharmaceutically acceptable carrier"
refers to any non-toxic amount of any substance suitable as a
vehicle for delivering a molecule or composition to a suitable in
vivo site of absorption. Examples of such carriers include, but are
not limited to water, phosphate buffered saline (PBS), Ringer's
solution, dextrose solution, serum-containing solutions, Hank's
solution and other aqueous physiologically-balanced solutions.
[0210] As used herein, a "pharmaceutically acceptable preservative"
includes but is not limited to a non-toxic amount of potassium
sorbate, methylparaben, propylparaben, benzoic acid and its salts,
other esters of parahydroxybenzoic acid such as butylparaben,
alcohols such as ethyl or benzyl alcohol, phenolic compounds such
as phenol, or quarternary compounds such as benzalkonium
chloride.
[0211] As used herein, a "coloring agent" provides coloration to
the composition or dosage form. Such coloring agents include, for
example, food grade dyes.
[0212] As used herein, the term "subject" refers to a mammal,
including but not limited to a dog, cat, horse, cow, pig, sheep,
goat, chicken, rodent, primate or human. Subjects include animals
such as house pets (e.g., dogs, cats, and the like), agricultural
stock subjects (e.g., cows, horses, pigs, chickens, etc.),
laboratory subjects (e.g., mice, rats, rabbits, etc.), but are not
so limited. The human subject may be a pediatric, such as for
example neonate, adult, or a geriatric subject. The human subject
may be of either gender.
[0213] As used herein, an "effective amount" or "therapeutically
effective amount" of a composition as described in some embodiments
herein can be a quantity sufficient to achieve a desired
therapeutic and/or prophylactic effect, for example, an amount
which results in the prevention of, or a decrease in the symptoms
associated with, a disease that is being treated. The amount of
composition administered to the subject, particularly one in need
of the composition, can depend on the type and severity of the
disease and on the characteristics of the individual, such as
general health, age, sex, body weight and tolerance to drugs. A
person skilled in the art will be able to determine appropriate
dosages depending on these and other factors. Typically, an
effective amount of the compositions described herein can be
sufficient for achieving a therapeutic or prophylactic effect.
[0214] In conjunction with the determination of a "therapeutically
effective amount" of a particular modality, it is within the
purview of the present invention to determine the presence of such
a "therapeutically effective amount" by determining a quantity of a
composition as described in some embodiments herein to modulate
biomarkers related to a particular disease state. Such biomarkers
may form a basis for surrogate endpoints for clinical trials,
provide a basis for elucidating a better understanding of disease
pathogenesis, and for quantifying the effectiveness of a
therapeutic modality to down-regulate an inflammatory response,
thereby quantifying disease progression.
[0215] Illustrative, albeit non-limiting biomarkers for several of
the disease states effectively treated by the compositions of the
present invention may include:
SCD
[0216] D-Dimer, to assess the coagulation rate, Total white blood
cells and platelets, markers of inflammatory state, Lactate
dehydrogenase (LDH), marker of intravascular hemolysis, Dense red
blood cell fraction, marker of red blood cell sickling,
Endothelin-1 (ET-1), soluble-VCAM-1 (sVCAM-1), soluble P-selectin
(sP-selectin), markers of endothelial activation, interleukins
(IL2, IL3, IL6, IL8 and IL10), urinary cysteinyl leucotriene E4
(28), and serum levels of prostaglandin-E2, CA 15-3, soluble CD40
ligand, HSP-70, ferritin, angiopoietin 1 and 2, stromal derived
factor 1, tumor necrosis factor-.alpha. and tumor necrosis factor
receptor-1.
NAFLD/NASH
[0217] CK18 M30, marker of NAFLD severity, IL-6, inflammatory
state, Urine isoprostane, Oxidative stress, Livers spectroscopy,
Fatty liver assessment
PSC
[0218] Calprotectin, Alkaline phosphatase--an elevated ALP is the
most common biochemical abnormality in PSC, which often prompts
clinicians to establish a diagnosis, reduction of ALP over time is
associated with a favorable prognosis, Aspartate to alanine
aminotransferase ratio: Aspartate aminotransferase (AST) and
alanine aminotransferase (ALT) are markers of hepatocellular injury
or necrosis, and elevation of AST or ALT is present in case of
liver damage, Bilirubin: Bilirubin is routinely used in clinical
practice as a measure of extrahepatic obstruction, Immunoglobulin
G4: Measure of inflammation, inflammation results in fibrosis and
deposition of connective tissue. PSC patients with elevated IgG4
have been shown to have a high prevalence of cirrhosis, and a
shorter time to liver transplant, suggesting a more severe liver
disease course.
[0219] It is contemplated to elucidate such biomarkers for any of
the disease states treated herein, and to determine a
therapeutically effective amount of a composition as it relates to
its ability to modulate the pathogenesis of said disease state.
[0220] The terms "dose unit," "unit dose," and "dosage unit," as
used herein, refer to a portion of a composition that contains an
effective amount of an active suitable for a single administration
to provide, or contribute to, a therapeutic effect. Such dosage
units may be administered one to a plurality (i.e., 1 to about 10,
1 to about 8, 1 to about 6, 1 to about 4 or 1 to about 2) of times
per day, or as many times as needed to elicit a therapeutic
response.
[0221] The term "food effect," as used herein, refers to a relative
difference in AUC (area under the curve), C.sub.max (maximum plasma
concentration), and/or T.sub.max (time to maximum concentration) of
an active substance, when said substance or a composition thereof,
such as a tablet, a capsule or a liquid, is administered orally to
a subject concomitantly with food or in a fed state as compared to
the same values when the same composition is administered in a
fasted state. The food effect, F, is calculated as:
F=(Y.sub.fed-Y.sub.fasted)/Y.sub.fasted
[0222] wherein Y.sub.fed and Y.sub.fasted are the found values of
AUC, C.sub.max, or T.sub.max in the fed and fasted state,
respectively. A food effect, F, is generally established when
F>1.
[0223] In general, the term "AUC" or "area under the plasma
concentration-time curve" is related to the total amount of an
active measurable in the systemic circulation following
administration of a single dose. The AUC is a mathematical and
visual representation of the aggregate amount of the active in the
systemic circulation over a given period of time. Changes in the
AUC need not necessarily reflect changes in the total amount of the
active absorbed but can reflect modifications in the kinetics of
distribution, metabolism and excretion. Accordingly, the term AUC
as used herein refers to the total amount of omega-3 fatty acids
(in either the triglyceride, ester, or free fatty acid form)
measurable in the systemic circulation following administration of
a single dose of any of the compositions described herein.
[0224] The term "T.sub.max" or "time of peak concentration" refers
to the period of time required to achieve peak plasma concentration
of an active after administration of a single dose. Accordingly,
the term T.sub.max as used herein refers to the period of time
required to achieve peak plasma concentration of omega-3 fatty acid
(in either the triglyceride, ester, or free fatty acid form) after
administration of a single dose of any of the compositions
described herein.
[0225] The term "C.sub.max" or "peak concentration" is the highest
concentration of an active achieved in the blood plasma.
Accordingly, the term C.sub.max as used herein refers to the
maximum concentration of omega-3 fatty acids (in either the
triglyceride, ester, or free fatty acid form) after administration
of a single dose of any of the compositions described herein.
[0226] The term "substantially independent of a food effect," or
"substantially free of food effect" as used herein, refers to a
significant elimination of the effect of food upon the absorption
(e.g., F is about 0), following oral administration, of any of the
compositions described herein. In other words, the bioavailability
of the omega-3 fatty acids (in either the triglyceride, ester, or
free fatty acid form), as measured by the logarithm-transformed
AUC.sub.0-.infin., is bioequivalent i.e., the 90% confidence
interval should be within the acceptance range of 80% to 125%
regardless of whether the compositions described herein are
administered with or without food. In certain embodiments, the
pharmacological effects of administration of compositions described
herein are substantially independent of a food effect.
[0227] The term "reduced food effect," as used herein, refers to a
substantial reduction in the effect of food upon the absorption,
following oral administration, of any of the compositions
described. In certain embodiments, the compositions described
herein have a reduced food effect.
[0228] The term "concomitantly with food" or "administration in the
fed state," as used herein, refers to administration from about 30
minutes before a meal to about 1 hour after a meal.
[0229] Various modes of treatment or prevention of medical
conditions as described herein are intended to mean "substantial"
or "substantially", which includes total but also less than total
treatment or prevention, and wherein some biologically or medically
relevant result is achieved. A subject, such as a human subject, in
need of treatment refers to a subject in need of treatment of a
defined disease state or in need of preventative treatment (i.e.,
prophylaxis) of such a disease state.
[0230] The use of numerical values in the various quantitative
values specified throughout this disclosure, unless expressly
indicated otherwise, are stated as approximations as though the
minimum and maximum values within the stated ranges were both
preceded by the word "about" unless expressly stated otherwise. In
this manner, equivalent variations from a stated value can be used
to achieve substantially the same results as the stated value.
Also, the disclosure of ranges is intended as a continuous range
including every value between the minimum and maximum values
recited as well as any ranges that can be formed by such values.
Also disclosed herein are any and all ratios (and ranges of any
such ratios) that can be formed by dividing a recited numeric value
into any other recited numeric value. Accordingly, the skilled
person will appreciate that many such ratios, ranges, and ranges of
ratios can be unambiguously derived from the numerical values
presented herein and in all instances such ratios, ranges, and
ranges of ratios represent various embodiments of the present
invention.
[0231] The term "functional food" as used herein means any edible
or drinkable foods or dietary components (e.g., infant formula,
juices, milk, yogurt, butter, margarine, baking products) that are
fortified or enhanced with any of the compositions described
herein. The functional food can be, e.g., solid, liquid, semisolid,
or a combination thereof. The term "functional food" also
encompasses edible and drinkable nutritional supplements.
[0232] The term "hydrophilic-lipophilic balance" or "HLB," as used
herein, refers to the relative affinity of a substance or
composition for aqueous and oily phases. HLB values can be
calculated based on methods and equations known to those of
ordinary skill in the art, such as those described in U.S. Pat. No.
5,585,192. Substances or compositions generally have an average HLB
of about 6 to about 20. Hydrophilic-lipophilic balance values can
be determined in a variety of the formulas or experimental methods
provided, for example, in U.S. Pat. No. 5,585,192.
[0233] The term "substantially pure" as used herein means at least
80% pure.
[0234] Pharmaceutical Compositions
[0235] In at least one embodiment, a composition is provided,
wherein the composition comprises at least one omega-3 fatty acid
and at least one surface active agent. In certain embodiments, the
at least one omega-3 fatty acid is selected from the group
consisting of hexadecatrienoic acid, .alpha.-linolenic acid,
stearidonic acid, eicosatrienoic acid, eicosapentaenoic acid,
heneicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic
acid, tetracosapentaenoic acid, and tetracosahexaenoic acid.
Certain embodiments comprise more than one omega-3 fatty acid,
which can be selected from the group consisting of hexadecatrienoic
acid, .alpha.-linolenic acid, stearidonic acid, eicosatrienoic
acid, eicosapentaenoic acid, heneicosapentaenoic acid,
docosapentaenoic acid, docosahexaenoic acid, tetracosapentaenoic
acid, and tetracosahexaenoic acid. The omega-3 fatty acid
components comprising the composition are present at, for example,
60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% (wt/wt) of the
composition. The omega-3 fatty acids used herein are typically in
the form of an oil.
[0236] In at least one embodiment, a composition is provided,
wherein the composition comprises at least one isomer of
docosapentaenoic acid (DPA) and at least one surface active agent,
wherein the omega-3 DPA isomer is
all-cis-7,10,13,16,19-docosapentaenoic acid (clupanodonic acid) or
the omega-6 DPA isomer is all-cis-4,7,10,13,16-docosapentaenoic
acid (osbond acid).
[0237] In certain embodiments, the compositions described herein
comprise DHA at, for example, from 50% to 99% (wt/wt) of the
compositions. For example, in certain embodiments, the DHA is
present at 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%
(wt/wt) of the compositions. In certain other embodiments, the DHA
is present at least, for example, 90% (wt/wt) of the compositions
described herein. In still other embodiments, the DHA is present
at, at least, for example, 95% (wt/wt) of the compositions
described herein.
[0238] In certain embodiments, the compositions described herein
comprise EPA at, for example, from 0% to 50% (wt/wt) of the
compositions. For example, in certain embodiments, the EPA is
present at <1% (wt/wt) of said compositions. In certain other
embodiments, the EPA can be present at, for example, 2%, 4%, 6%,
8%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 32%,
34%, 36%, 38%, 40%, 42%, 44%, 46%, 48% or 50% (wt/wt) of said
compositions. In still other embodiments, the EPA can be present at
<5% (wt/wt) of said compositions. For example, in certain
embodiments, the EPA can be present at 0%, 0.1%, 0.2%, 0.3%, 0.4%,
0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, or
4.5% (wt/wt) of said compositions.
[0239] In certain embodiments, the compositions described herein
comprise an omega-3 fatty acid other than DHA and EPA at, for
example, from >0% to 10% (wt/wt) of said compositions. For
example, said omega-3 fatty acid other than DHA and EPA can be
present, in certain embodiments at 0.1%, 0.2%, 0.3%, 0.4%, 0.5%,
0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%,
5.5%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10% (wt/wt) of said
compositions. In such embodiments, the omega-3 fatty acid other
than EPA and DHA can be selected from the group consisting of
hexadecatrienoic acid, .alpha.-linolenic acid, stearidonic acid,
eicosatrienoic acid, heneicosapentaenoic acid, docosapentaenoic
acid, tetracosapentaenoic acid, and tetracosahexaenoic acid. In
certain such embodiments, the third fatty acid other than DHA and
EPA may be an omega-6 fatty acid, for example arachidonic acid
(ARA), linoleic acid (LA), gamma-linolenic acid (GLA) or the
omega-6 isomer of DPA.
[0240] In certain embodiments, the compositions described herein
comprise 90%, 5%, and 5% (wt/wt) of said compositions DHA, ARA, and
EPA respectively.
[0241] In certain embodiments, the compositions described herein
comprise 95%, 5%, and <1% (wt/wt) of said compositions DHA, ARA,
and EPA respectively.
[0242] The omega-3 fatty acid oils used in the manufacture of the
compositions described herein are substantially pure. In some
embodiments, the omega-3 fatty acid oils are at least, for example,
90% pure; and the remainder of the omega-3 fatty acids in the oil
can comprise a mixture of other omega-3 fatty acids of which one
omega-3 fatty acid may predominate the mixture. For example, if a
composition comprises DHA oil, the DHA oil can be at least 90% DHA;
and the remainder can comprise a mixture of omega-3 fatty acids, of
which EPA can be the predominant omega-3 fatty acid and can be
present at 5% of said composition. Accordingly, in omega-3 fatty
acid oils, the ratio of the amount of the substantially pure
omega-3 fatty acid, the first omega-3 fatty acid, to the
predominant omega-3 fatty acid in the remaining fraction of the
oil, the second omega-3 fatty acid, is 1:X, where 0<X<0.3 or
0.5<X<1. Thus, in certain embodiments, X can be, for example,
0.001, 0.0015, 0.002, 0.0025, 0.0055, 0.006, 0.0065, 0.007, 0.0075,
0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03,
0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08,
0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3, 0.55, 0.6, 0.65,
0.7, 0.75, or 0.8.
[0243] Thus, in at least one embodiment, the composition described
herein comprises at least one surface active agent and a first and
second omega-3 fatty acid, wherein the first omega-3 fatty acid is
DHA and the second omega-3 fatty acid EPA, wherein the ratio of the
amount of DHA:EPA is 1:X, where 0<X<0.3 or 0.5<X<1. In
certain other embodiments, X can be, for example, 0.001, 0.0015,
0.002, 0.0025, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085,
0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045,
0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095,
0.1, 0.15, 0.2, 0.25, 0.55, 0.6, 0.65, 0.7, 0.75, or 0.8.
[0244] In other embodiments, the compositions described herein
comprise at least one surface active agent and a first and second
omega-3 fatty acid, wherein the first omega-3 fatty acid is EPA and
the second omega-3 fatty acid is DHA, wherein the ratio of the
amount of EPA:DHA is 1:X, where 0<X<0.3 or 0.5<X<1. In
certain other embodiments, X can be, for example, 0.001, 0.0015,
0.002, 0.0025, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085,
0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.055, 0.06, 0.065,
0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.55,
0.6, 0.65, 0.7, 0.75, or 0.8.
[0245] In certain embodiments, the compositions described herein
can optionally comprise a third omega fatty acid (either in the
triglyceride, ester, or free fatty acid form), wherein the third
omega fatty acid is different from the first and second omega fatty
acids. In such compositions, the ratio of the amount of the first
omega fatty acid to the second and third omega fatty acids is 1:X,
where X is the combined amount of the second and third omega fatty
acids and X is 0<X<1. For example, if a composition comprises
DHA oil, the DHA oil can be at least, for example, 90% DHA (the
first omega fatty acid); and the remaining 10% can comprise a
mixture of omega fatty acids, of which EPA can be the predominant
omega fatty acid (second omega fatty acid) and can be present at 5%
of said composition. The third omega fatty acid is selected from
the group consisting of hexadecatrienoic acid, .alpha.-linolenic
acid, stearidonic acid, eicosatrienoic acid, heneicosapentaenoic
acid, docosapentaenoic acid, tetracosapentaenoic acid, and
tetracosahexaenoic acid. In certain embodiments the third omega
acid may be an omega-6 fatty acid. Accordingly, in such
embodiments, if the first omega fatty acid is DHA and the second
omega fatty acid is EPA, The ratio of the amount of DHA:(EPA+third
omega fatty acid) is 1:X, where X is the amount of EPA and the
amount of the third omega fatty acid, and where 0<X<1. In
certain embodiments, X can be, for example, 0.001, 0.0015, 0.002,
0.0025, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009,
0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05,
0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1,
0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7,
0.75, 0.8, 0.85, 0.9, or 0.95. In certain embodiments, the third
fatty acid can be arachidonic acid (ARA) (either in the
triglyceride, ester, or free fatty acid form). In certain
embodiments, the ARA is in the ethyl ester form.
[0246] In some embodiments, the omega-3 or omega-6 fatty acid
compositions described herein are in either the triglyceride,
ester, or free fatty acid form. Typically, but not necessarily, the
omega-3 or omega-6 fatty acid is in the ester form, particularly in
the ethyl ester form.
[0247] In certain embodiments, the compositions comprise at least
one surface active agent and a first and second omega-3 fatty acid,
wherein the first omega-3 fatty acid (in either the triglyceride,
ester, or free fatty acid form) is omega-3 docosapentaenoic acid
(omega-3 DPA) and the second omega-3 fatty acid (in either the
triglyceride, ester, or free fatty acid form) is selected from the
group consisting of docosahexaenoic acid (DHA), eicosapentaenoic
acid (EPA), hexadecatrienoic acid, .alpha.-linolenic acid,
stearidonic acid, eicosatrienoic acid, eicosapentaenoic acid,
heneicosapentaenoic acid, docosapentaenoic acid,
tetracosapentaenoic acid, and tetracosahexaenoic acid and
combinations thereof.
[0248] In certain embodiments, the compositions comprise at least
one surface active agent, an omega-6 fatty acid, and an omega-3
fatty acid, wherein the omega-6 fatty acid (in either the
triglyceride, ester, or free fatty acid form) is for example
arachidonic acid (ARA), linoleic acid (LA), gamma-linolenic acid
(GLA) or the omega-6 isomer of DPA, and the second omega-3 fatty
acid (in either the triglyceride, ester, or free fatty acid form)
is selected from the group consisting of omega-3 docosapentaenoic
acid (omega-3 DPA), docosahexaenoic acid (DHA), eicosapentaenoic
acid (EPA), hexadecatrienoic acid, .alpha.-linolenic acid,
stearidonic acid, eicosatrienoic acid, eicosapentaenoic acid,
heneicosapentaenoic acid, docosapentaenoic acid,
tetracosapentaenoic acid, and tetracosahexaenoic acid and
combinations thereof.
[0249] In certain embodiments the omega-3 fatty acid (in either the
triglyceride, ester, or free fatty acid form) may be DHA, for
example substantially pure DHA, in combination with arachidonic
acid in a ratio of DHA to arachidonic acid of 1:1 to 20:1. In
certain embodiments, .alpha.-linolenic acid may be included for its
anti-inflammatory properties.
[0250] In certain embodiments, the compositions described herein
are free of free fatty acids.
[0251] In certain embodiments, the compositions described herein
are free of a second medicament.
[0252] In certain embodiments, the at least one surface active
agent is selected from the group consisting of nonionic surface
active agents, cationic surface active agents, anionic surface
active agents, zwitterionic surface active agents, or combinations
thereof.
[0253] In some embodiments, the compositions include one or more
non-ionic surface active agents. Non-ionic surface active agents
generally have a hydrophobic group and a reactive hydrogen atom,
for example aliphatic alcohols, acids, amides and alkyl phenols,
with alkylene oxides, especially ethylene oxide either alone or in
combination with propylene oxide. Examples of nonionic surfactant
compounds include, but are not limited to, polyoxyethylene glycol
sorbitan alkyl esters, block copolymers of polyethylene glycol and
polypropylene glycol, ethylene glycol fatty acid esters,
poly(ethylene glycol) fatty acid esters, propylene glycol fatty
acid esters, poly(propylene glycol) fatty acid esters, glycol fatty
acid esters, trimethylolpropane fatty acid esters, pentaerythritol
fatty acid esters, glucoside derivatives, glycerin alkyl ether
fatty acid esters, trimethylolpropane oxyethylene alkyl ethers,
fatty acid amides, alkylolamides, alkylamine oxides, lanolin and
its derivatives, castor oil derivatives, hardened castor oil
derivatives, sterols and its derivatives, polyoxyethylene alkyl
ethers, polyoxyethylene alkyl allyl ethers, polyoxyethylene
alkylamine, polyoxyethylene fatty acid amides, polyoxyethylene
alkylolamides, polyoxyethylene diethanolamine fatty acid esters,
polyoxyethylene trimethylolpropane fatty acid esters,
polyoxyethylene alkyl ether fatty acid esters, polyoxyethylene
polyoxypropylene glycols, polyoxyethylene polyoxypropylene alkyl
ethers, polyoxyethylene polyoxypropylene polyhydric alcohol ethers,
glycerin fatty acid esters, polyglycerin fatty acid esters,
polyoxyethylene glycerin fatty acid esters, sorbitan fatty acid
esters, polyoxyethylene sorbitan fatty acid esters, sucrose fatty
acid esters, or combinations thereof.
[0254] In certain embodiments, the surface active agents comprise
polyoxyethylene glycol sorbitan alkyl esters, block copolymers of
polyethylene glycol and polypropylene glycol, or combinations
thereof.
[0255] Examples of polyoxyethylene glycol sorbitan alkyl esters are
typically the polysorbates. Polysorbates are a class of oily
liquids derived from PEG-ylated sorbitan (a derivative of sorbitol)
esterified with fatty acids. Common brand names for polysorbates
include Tween.RTM.. Tween.RTM.-20, Tween.RTM.-40, Tween.RTM.-60 and
Tween.RTM.-80, for example, are available from AkzoNobel
(Strawinskylaan 2555 1077 ZZ, Amsterdam, the Netherlands).
Exemplary polysorbates include polysorbate 20 (polyoxyethylene (20)
sorbitan monolaurate), polysorbate 40 (polyoxyethylene (20)
sorbitan monopalmitate), polysorbate 60 (polyoxyethylene (20)
sorbitan monostearate), and polysorbate 80 (polyoxyethylene (20)
sorbitan monooleate).
[0256] Examples of block copolymers of polyethylene glycol and
polypropylene glycol include the poloxamers. Poloxamers are
nonionic triblock copolymers composed of a central hydrophobic
chain of polyoxypropylene (poly(propylene oxide)) flanked by two
hydrophilic chains of polyoxyethylene (poly(ethylene oxide)).
Certain poloxamers, such as those listed herein, are also known by
the trade names Pluronic.RTM. available from suppliers such as BASF
AG (Ludwigshafen, Germany). Because the lengths of the polymer
blocks can be customized, many different poloxamers exist that have
slightly different properties. Further exemplary Pluronic.RTM.
poloxamers include, but are not limited to Pluronic.RTM. 10R5,
Pluronic.RTM. 17R2, Pluronic.RTM. 17R4, Pluronic.RTM. 25R2,
Pluronic.RTM. 25R4, Pluronic.RTM. 31R1, Pluronic.RTM. F 108 Cast
Solid Surfacta, Pluronic.RTM. F 108 NF, Pluronic.RTM. F 108
Pastille, Pluronic.RTM. F 108 Prill, Pluronic.RTM. F 108NF Prill
Poloxamer 338, Pluronic.RTM. F 127, Pluronic.RTM. F 127 Prill,
Pluronic.RTM. F 127 NF, Pluronic.RTM. F 127 NF 500 BHT Prill,
Pluronic.RTM. F 127 NF Prill Poloxamer 407, Pluronic.RTM. F 38,
Pluronic.RTM. F 38 Pastille, Pluronic.RTM. F 68, Pluronic.RTM. F 68
Pastille, Pluronic.RTM. F 68 LF Pastille, Pluronic.RTM. F 68 NF,
Pluronic.RTM. F 68 NF Prill Poloxamer 188, Pluronic.RTM. F 68
Prill, Pluronic.RTM. F 68 Prill, Pluronic.RTM. F 77, Pluronic.RTM.
F 77 Micropastille, Pluronic.RTM. F 87, Pluronic.RTM. F 87 NF,
Pluronic.RTM. F 87 NF Prill Poloxamer 237, Pluronic.RTM. F 87
Prill, Pluronic.RTM. F 88, Pluronic.RTM. F 88 Pastille,
Pluronic.RTM. F 88 Prill, Pluronic.RTM. F 98, Pluronic.RTM. F 88
Prill, Pluronic.RTM. F 98, Pluronic.RTM. F 98 Prill, Pluronic.RTM.
L 10, Pluronic.RTM. L 101, Pluronic.RTM. L 121, Pluronic.RTM. L 31,
Pluronic.RTM. L 35, Pluronic.RTM. L 43, Pluronic.RTM. L 44,
Pluronic.RTM. L 61, Pluronic.RTM. L 62, Pluronic.RTM. L 62 LF,
Pluronic.RTM. L 62D, Pluronic.RTM. L 64, Pluronic.RTM. L 81,
Pluronic.RTM. L 92, Pluronic.RTM. L44 NF INH surfactant Poloxamer
124, Pluronic.RTM. N 3, Pluronic.RTM. P 103, Pluronic.RTM. P 104,
Pluronic.RTM. P 105, Pluronic.RTM. P 123 Surfactant, Pluronic.RTM.
P 65, Pluronic.RTM. P 84, Pluronic.RTM. P 85, or combinations
thereof.
[0257] In certain embodiments, the at least one surface active
agent comprises a polysorbate present from, for example, 15%
(wt/wt) to 35% (wt/wt) of the composition.
[0258] In certain embodiments, the polysorbate is present at, for
example, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%,
26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, or 35% (wt/wt) of said
composition.
[0259] In certain embodiments, the polysorbate is present at, for
example, 31% (wt/wt) of the composition.
[0260] In certain embodiments, the polysorbate is polysorbate 20,
polysorbate 60, polysorbate 80 or a combination thereof. Typically,
but not necessarily, the polysorbate is polysorbate 80.
[0261] In certain other embodiments, the at least one surface
active agent comprises a poloxamer present from, for example, 0.1%
(wt/wt) to 5% (wt/wt) of the composition.
[0262] In certain embodiments, the poloxamer is present at, for
example, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%,
1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, or 5% (wt/wt) of said
composition.
[0263] In certain embodiments, the poloxamer is present at, for
example, 0.7% (wt/wt) of the composition.
[0264] In certain embodiments, the poloxamer is Pluronic.RTM. F87
[(HO(C.sub.2H.sub.4O).sub.64(C.sub.3H.sub.6O).sub.37(C.sub.2H.sub.4O).sub-
.64H], which is poloxamer 237.
[0265] In certain embodiments, the compositions described herein
comprise a combination of polysorbate 80 and the poloxamer
Pluronic.RTM. F87
[(HO(C.sub.2H.sub.4O).sub.64(C.sub.3H.sub.6O).sub.37(C.sub.2H.sub.4O).sub-
.64H], which is poloxamer 237.
[0266] In certain embodiments, the at least one surface active
agent comprises a combination of nonionic surfactants. For example,
the combination of nonionic surfactants can comprise a combination
of a polysorbate and a poloxamer totaling at least, for example,
30% (wt/wt) of said composition. In certain embodiments, the
polysorbate is polysorbate 80 and the poloxamer is poloxamer 237
(Pluronic.RTM. F87). In certain embodiments, the polysorbate 80 is
present at, about 31% (wt/wt) and the poloxamer 237 is present at,
about 0.7% (wt/wt) of said composition.
[0267] In some embodiments, the compositions include one or more
anionic surface active agents. Exemplary "anionic surface active
agents" include, but are not limited to, N-acyl-L-glutamic acid
diethanolamine, N-acyl-L-glutamic acid triethanolamine, sodium
N-acyl-L-glutamate, sodium alkanesulfonate, ammonium alkyl (C12,
C14, C16) sulfate, alkyl (C11, C13, C15) sulfuric acid
triethanolamine, alkyl (C11, C13, C15) sulfuric acid
triethanolamine, alkyl (C12 to C14) sulfuric acid triethanolamine,
liquid alkylsulfuric acid triethanolamine, sodium alkyl (C12, C13)
sulfate, liquid sodium alkylsulfate, sodium isoethionate, sodium
lacto-isostearate, disodium undecylenoylamido ethyl sulfosuccinate,
triethanolamine sulfooleate, sodium sulfooleate, disodium oleamide
sulfosuccinate, potassium oleate, sodium oleate, morpholine oleate,
oleoyl sarcosine, oleoyl methyltaurine sodium salt,
potassium-containing soap base, liquid base for potassium soap,
potassium soap, carboxylated polyoxyethylene tridodecyl ether,
sodium salt (3 ethyle oxide "E.O.") of carboxylated polyoxyethylene
tridodecyl ether, triethanolamine N-hydrogenated tallow
fatty-acyl-L-glutamate, sodium N-hydrogenated tallow
fatty-acyl-L-glutamate, sodium hydrogenated coconut fatty acid
glyceryl sulfate, sodium diundecylenoylamido ethyl sulfosuccinate,
sodium stearyl sulfate, potassium stearate, triethanolamine
stearate, sodium stearate, sodium N-stearoyl-L-glutamate, disodium
stearoyl-L-glutamate, stearoyl methyltaurine sodium salt, sodium
dioctyl sulfosuccinate, liquid sodium dioctyl sulfosuccinate,
liquid disodium polyoxyethylene monooleylamido sulfosuccinate (2
E.O.), disodium polyoxyethylene lauroyl ethanolamide sulfosuccinate
(5 E.O.), disodium lauryl sulfosuccinate, diethanolamide cetyl
sulfate, sodium cetyl sulfate, soap base, sodium cetostearyl
sulfate, triethanolamine tridecyl sulfate, potassium palmitate,
sodium palmitate, palmitoyl methyltaurine sodium salt, liquid
castor oil fatty acid sodium salt (30%), ammonium polyoxyethylene
alkyl ether sulfate (3 E.O.), liquid diethanolamine polyoxyethylene
alkyl (C12, C13) ether sulfate, liquid triethanolamine
polyoxyethylene alkyl ether sulfate (3 E.O.), triethanolamine
polyoxyethylene alkyl (C11, C13, C15) ether sulfate (1 E.O.),
triethanolamine polyoxyethylene alkyl (C12, C13) ether sulfate (3
E.O.), liquid sodium polyoxyethylene alkyl ether sulfate (3 E.O.),
sodium polyoxyethylene alkyl (C11, C13, C15) ether sulfate (1
E.O.), sodium polyoxyethylene alkyl (C11 to C15) ether sulfate (3
E.O.), sodium polyoxyethylene alkyl (C12, C13) ether sulfate (3
E.O.), sodium polyoxyethylene alkyl (C12 to C14) ether sulfate (3
E.O.), sodium polyoxyethylene alkyl (C12 to C15) ether sulfate (3
E.O.), disodium polyoxyethylene alkyl (C12 to C14) sulfosuccinate
(7 E.O.), sodium polyoxyethylene undecyl ether sulfate, liquid
sodium polyoxyethylene octyl phenyl ether sulfate, ammonium
polyoxyethylene oleyl ether sulfate, disodium polyoxyethylene
lauryl sulfosuccinate, sodium polyoxyethylene nonyl phenyl ether
sulfate, sodium polyoxyethylene pentadecyl ether sulfate,
triethanolamine polyoxyethylene myristyl ether sulfate, sodium
polyoxyethylene myristyl ether sulfate, sodium polyoxyethylene
myristyl ether sulfate (3 E.O.), liquid sodium polyoxyethylene
lauryl ether acetate (16 E.O.), ammonium polyoxyethylene lauryl
ether sulfate (2 E.O.), triethanolamine polyoxyethylene lauryl
ether sulfate, sodium polyoxyethylene lauryl ether sulfate,
diethanolamine myristyl sulfate, sodium myristyl sulfate, potassium
myristyl sulfate, sodium N-myristoyl-L-glutamate, sodium
myristoylmethylaminoacetate, liquid myristoyl methyl-alanine sodium
salt, myristoyl methyltaurine sodium salt, medicinal soaps,
triethanolamine/magnesium coco alkyl sulfate, triethanolamine
N-coconut oil fatty-acyl-L-glutamate, sodium N-coconut oil
fatty-acyl-L-glutamate, sodium coconut oil fatty acid ethyl ester
sulfonate, coconut oil fatty acid potassium salt, liquid coconut
oil fatty acid potassium salt, sodium N-coconut oil
fatty/hydrogenated fatty-acyl-L-glutamate, coconut oil fatty acid
sarcosine, coconut oil fatty acid sarcosine triethanolamine salt,
coconut oil fatty acid sarcosine sodium salt, coconut oil fatty
acid triethanolamine salt, liquid triethanolamine salt of coconut
oil fatty acid, coconut oil fatty acid sodium salt, coconut oil
fatty acid methyl alanine sodium salt, liquid coconut oil fatty
acid methyl alanine sodium salt, coconut oil fatty acid
methyltaurine potassium salt, coconut oil fatty acid methyltaurine
sodium salt, sodium laurylamino dipropionate, liquid sodium
laurylamino dipropionate (30%), sodium lauryl sulfoacetate; sodium
lauryl benzenesulfonate, lauryl sulfate, ammonium lauryl sulate,
potassium lauryl sulfate, diethanolamine lauryl sulfate,
triethanolamine lauryl sulfate, sodium lauryl sulfate, magnesium
lauryl sulfate, monoethanolainine lauryl sulfate, potassium
laurate, lauric acid triethanolamine, liquid lauric acid
triethanolamine, sodium laurate, lauric acid/myristic acid
triethanolamine, lauroyl-L-glutamic acid triethanolamine, sodium
N-lauroyl-L-glutamate, lauroyl sarcosine, lauroyl sarcosine
potassium, liquid lauroyl sarcosine triethanolamine salt, lauroyl
sarcosine sodium, liquid lauroyl methyl-.beta.-alanine sodium salt,
lauroyl methyltaurine sodium salt, liquid lauroyl methyltaurine
sodium salt, or combinations thereof.
[0268] In certain embodiments, said anionic surfactant(s) comprise
from, for example, about 0.05% (wt/wt) to about 25% (wt/wt) of said
composition. In certain embodiments, said anionic surfactant(s)
comprise from, for example, about 0.05% (wt/wt) to about 15%
(wt/wt) of said composition. In certain embodiments, said anionic
surfactant(s) comprise, for example, 0.05% (wt/wt) to 5% (wt/wt) of
said composition. In certain embodiments, said anionic
surfactant(s) comprise, for example, 0.5% (wt/wt) to 3% (wt/wt) of
said composition. In certain embodiments, said anionic
surfactant(s) comprise, for example, 0.7% (wt/wt) of said
composition. In certain embodiments, said anionic surfactant(s)
comprise sodium lauryl sulfate.
[0269] In some embodiments, the compositions include additional
surface active agents such as the zwitterionic and cationic surface
active agents. Examples of such surface active agents include, but
are not limited to the bile acids (e.g., cholic acid,
chenodeoxycholic acid, glycocholic acid, glycodeoxycholic acid,
taurocholic acid, taurochenodeoxycholic acid, taurolithocholic
acid, deoxycholic acid, lithocholic acid, and ursodeoxycholic acid
and salts thereof, e.g., sodium, potassium, lithium), natural
emulsifiers (e.g. acacia, agar, alginic acid, sodium alginate,
tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg
yolk, casein, wool fat, cholesterol, wax, and lecithin), long chain
amino acid derivatives, high molecular weight alcohols (e.g.
stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin
monostearate, ethylene glycol distearate, glyceryl monostearate,
and propylene glycol monostearate, polyvinyl alcohol), carbomers
(e.g. carboxy polymethylene, polyacrylic acid, acrylic acid
polymer, and carboxyvinyl polymer), carrageenan, cellulosic
derivatives (e.g. carboxymethylcellulose sodium, powdered
cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose,
hydroxypropyl methylcellulose, methyl cellulose), polyoxyethylene
esters (e.g. polyoxyethylene monostearate [Myrj 45],
polyoxyethylene hydrogenated castor oil, polyethoxylated castor
oil, polyoxymethylene stearate, and Solutol), sucrose fatty acid
esters, polyethylene glycol fatty acid esters (e.g. Cremophor),
polyoxyethylene ethers, (e.g. polyoxyethylene lauryl ether [Brij
30]), poly(vinyl-pyrrolidone), diethylene glycol monolaurate,
triethanolamine oleate, sodium oleate, potassium oleate, ethyl
oleate, oleic acid, ethyllaurate, sodium lauryl sulfate,
cetrimonium bromide, cetylpyridinium chloride, benzalkonium
chloride, docusate sodium or combinations thereof.
[0270] Without being bound to any particular theory, it is believed
that the compositions described herein will increase fatty acid
absorption in patients with fat malabsorption syndromes and other
disease states as described herein. This is because the
compositions described herein comprising the omega-3 fatty acid(s)
and the at least one surface active agent self-micellize in a
manner similar to that in the body. When mammals eat, micelles are
naturally formed around the fats ingested. These natural micelles
form in the presence of bile regardless of the pH in the stomach or
intestines. Natural micelles form around fats and digestive
enzymes, entrapping both. Within the micelles, the enzymes quickly
cleave the triglycerides or fatty acid esters into free fatty acids
and monoglycerides. Natural micelles fuse with the cell membrane
when they come in contact with the brush barrier of the intestinal
wall, delivering their cargo of free fatty acids and
monoglycerides, among other components and compounds, into the
cells lining the intestine and are absorbed. Natural micelles that
form around omega-3 fatty acids have two characteristics that limit
their effectiveness. Firstly, given the formation of natural
micelles require the food-dependent release of bile salts, natural
micelles form only when food is present. Secondly, since natural
micelles require fats from the consumed food in order to form the
outer shell of the micelle, the amount of lipids transported into
the intestinal wall is directly correlated with the amount of fat
co-consumption. As a result, unless a lipophilic compound is
co-administered with food, very few natural micelles form and,
unless that food contains a sufficient amount of fat, very few
natural micelles form. It then follows that the lower the natural
micelle formation, the less efficient the natural process is at
delivering said lipophilic compound, among other components and
compounds, in micellar form, to the intestinal lumen.
[0271] In contrast to natural micelles, the formation of artificial
micelles has hitherto been very sensitive to the pH of the liquid
in which they are being created and to the presence of other
substances in that liquid. In addition, artificial micelles are in
general either fragile and break easily or are too rigid and never
release their cargo, making it difficult for them to deliver
adequate amounts of free fatty acids and monoglycerides to the
intestinal lining. The micelles formed by the compositions
described herein, however, overcome the deficiencies of both
artificial and natural human micelles in the delivery of omega-3
fatty acids, while at the same time mimicking some of the
characteristics of natural human micelles.
[0272] "Emulsion" is a very broad term describing particles that
may be amorphous, liquid-crystalline, bilayer, monolayer or any
mixture of these structures. An emulsion is a more general term
that describes a dispersed phase in a continuous phase that is
commonly seen as oil in water (o/w), water in oil (w/o) or even
more complex mixtures such as water in oil in water (w/o/w).
Emulsion particle sizes can vary widely from 10 nanometers to 100
micrometers.
[0273] Solutions in lipid vehicles containing surfactants, which
constitute self-emulsifying drug delivery systems (SEDDS), and
which effect spontaneous emulsification upon contact of the lipid
with fluids in the gastrointestinal tract (GI tract), are known in
the art, and are classified as "self-emulsified", "self-micro", and
"self-nano", drug delivery systems (herein referred to as "SEDDS,"
"SMEDDS," and "SNEDDS," respectively). The SEDDS, SMEDDS, and
SNEDDS are emulsions. These emulsions have the sole purpose of
dispersing lipids, such as oils, into small droplets in order to
increase the surface area available to interact with the body's
natural bile salts, pancreatic lipases, co-lipase, and
phospholipase A2.
[0274] SEDDS are formulated with mixtures of lipid vehicles,
non-ionic surfactants and a drug in the absence of water, and are
assumed to exist as transparent isotropic solutions. These systems
have a unique property--they are able to self-emulsify rapidly in
the gastrointestinal fluids, forming fine oil-in-water emulsions
(droplet size diameter <300 nm) under gentle agitation provided
by gastrointestinal motion. SEDDS are commonly suitable for oral
delivery in soft and hard gelatin or hard hydroxypropyl
methylcellulose (HPMC) capsules.
[0275] SMEDDS are defined as isotropic mixtures of an oil,
surfactant, co-surfactant (or solubilizer) and a drug. Such systems
form fine oil-in-water microemulsions under gentle agitation
provided by digestive motility of the stomach and intestine
following dilution by the aqueous phase in vivo. SMEDDS are
distinguished from SEDDS by smaller emulsion droplets produced on
dilution, resulting in a transparent or translucent stable
dispersion. Mean droplet size after dilution is <100 nm in the
case of SMEDDS or <300 nm in the case of SEDDS. SMEDDS generally
contain relatively high concentrations of surfactant (typically 30
to 60%, m/m), and optionally also hydrophilic co-solvents (e.g.,
propylene glycol, polyethylene glycols). They are often described
as microemulsion pre-concentrates because the microemulsion is
formed on dilution in aqueous media.
[0276] SNEDDS are defined as self-nanoemulsifying drug delivery
systems that spontaneously form transparent to opalescent
oil-in-water dispersions of approximately 200 nm in size upon
dilution with water under gentle stirring.
[0277] As these various emulsions move through the GI tract various
processes occur. First the components are dispersed to form lipid
droplets, or emulsion droplets. This is followed by lipolysis and
solubilization of the digestion products by bile acids, which may
then form colloidal mixed micelles. It is believed that the drug
then partitions from the emulsion oil droplets and bile salt mixed
micelles to be absorbed by the mucosal cells of the intestinal
wall.
[0278] Thus, because SEDDS, SMEDDS, and SNEDDS leverage the body's
natural process to be efficient, they also necessarily rely on the
natural processes to be effective. This is particularly important
because in order to form micelles, SEDDS, SMEDDS, and SNEDDS
require the presence of bile salts. Since bile salts are emitted
into the GI tract in mammals as a response to food consumption,
then it follows that SEDDS SMEDDS, and SNEDDS are not able to use
bile salts to form micelles in the GI tract unless food intake has
induced the introduction of bile salts into the GI tract.
[0279] On the contrary, the present invention is directed toward a
Self-Micellizing Drug Delivery System (SMDDS). This system is
differentiated from the SEDDS, SMEDDS and SNEDDS as described
above, in that the particularly described combination of surface
active agent (polysorbate and poloxamer 237) in the relative
amounts and overall concentrations as herein described, are unique
in their ability to directly and spontaneously form stable micelles
in a size range of about 1 to about 10 micron without relying on
the presence of bile salts. This represents a distinct and unique
advantage of the present invention, in that, unlike SEDDS, SMEDDS,
and SNEDDS, the present invention is able to deliver lipids through
the intestinal wall by way of a plurality of spontaneously formed
stable micelles. Unlike SEDDS, SMEDDS, and SNEDDS, which function
to disperse lipids to increase surface area, the present invention
encapsulates the lipids with surface active agents, thus bringing
the lipids together, whereby predictable and repeatable
enhancements in bioavailability are realized, absent any food
effect.
[0280] As opposed to an emulsion, a micelle is a particle of
colloidal dimensions that exists in equilibrium with molecules or
ions in a solution from which it is formed. Micelles are a
particular type of particle (oil particle in water) which form a
structure wherein the hydrophilic component is external and the
hydrophobic component is internal. More specifically, micelles are
usually formed from single chain lipids and surfactants, and a
micelle is always a monolayer particle with the hydrophilic head
facing the aqueous phase and the hydrophobic tail facing the oil
phase.
[0281] One method of determining a human's normal weight range is
to assess the subject's Basal Metabolic Rate (BMR), which takes
into account a subject's height, weight, age and gender. BMR
determinations are well known in the art. Accordingly, it is
believed, without being bound to any particular theory, that the
compositions or a plurality of micelles pre-formed therefrom
described herein will lower a human's weight, if the human is
overweight, to within the normal range as determined by the human's
BMR. Conversely, it is believed, without being bound to any
particular theory, that the compositions or a plurality of micelles
pre-formed therefrom described herein will increase a human's
weight, if the human is underweight, malnourished, or is suffering
from a malabsorption syndrome, to within the normal range as
determined by the human's BMR.
[0282] It is known in the art that food intake in a subject having
a normal weight stimulates adipose tissue to increase leptin
levels. Leptin is a hormone that tells the brain it is full and
increases metabolism. Thus, in starvation mode there is a decrease
in leptin, telling the brain it is hungry, which in turn stimulates
food intake and decreases energy expenditure. In an obese subject,
however, the brain is resistant to leptin signaling despite leptin
levels being high, thus, the subject believes it is not satiated
and continues eating and gaining weight.
[0283] Omega-3 fatty acids are known to decrease circulating leptin
levels. Accordingly, without being bound to any particular theory,
it is believed that the enhanced absorption of omega-3 fatty acids
provided by the compositions or a plurality of micelles pre-formed
therefrom described herein will regulate leptin levels and signal a
subject's brain accordingly to either decrease leptin levels if the
subject is overweight or increase leptin levels if the subject is
underweight, malnourished, or is suffering from a malabsorption
syndrome. This increase or decrease in metabolism, as the case may
be, is reflected by an increase or decrease in circulating free
fatty acids when compared to circulating free fatty acid levels at
the start of the dosing regimen.
[0284] Without being bound to any particular theory, it is believed
that the self-micellizing compositions described herein will
regulate metabolism in a subject administered said compositions to
facilitate returning the subject's weight to within a normal range
for that particular subject.
[0285] Accordingly, in at least one embodiment of the present
invention, a method is disclosed for administering self-micellizing
compositions described herein to facilitate a subject's weight
gain, loss, or return to normal weight, or combinations thereof
over time.
[0286] In at least one embodiment of the present invention, a
composition is disclosed which can be used to increase the weight
of a mammal, such as livestock.
[0287] Also disclosed herein in at least one embodiment of the
invention is a method for administering a composition as provided
herein to increase the weight of a mammal, such as livestock.
[0288] In at least one embodiment, the compositions described
herein comprising the omega-3 fatty acid(s) and the at least one
surface active agent, for example the combination of polysorbate 80
and poloxamer 237, spontaneously form micelles when encountering
aqueous liquids. These micelles form regardless of the pH or the
nature and concentration of other suspended materials in the
liquid, do not require bile to form, and form micelles that remain
stable almost indefinitely. The micelles form whether or not food
is present or whether food that is present is high or low in fat.
Like human micelles, the micelles formed by the compositions
described herein form around omega-3 fatty acids and entrap both
fats and enzymes, allowing rapid digestion and the formation of
free fatty acids and monoglycerides. Without being held to any one
theory, it is believed that the micelles formed by the compositions
described herein are similar in size to human micelles and share
the ability to rupture at the intestinal brush barrier. Without
being held to any one theory, it is believed that either by
friction or a chemical reaction, the micelle ruptures and spills
its contents onto the lining of the intestines and the intestines
absorb the omega-3 fatty acids.
[0289] Accordingly, it is believed that compositions described
herein can overcome one of the primary challenges of malabsorption
syndromes, such as SBS i.e., the delivery of adequate amount of
free fatty acids to the bloodstream of patients suffering from the
hepatic complications of SBS without exacerbating liver
dysfunction, as does TPN treatment, or triggering fat malabsorption
or dumping. This would also allow for more consistent dosing and
thus produce more reliable results.
[0290] In certain embodiments, the compositions described herein
self-micellize in an aqueous medium. The aqueous medium can
include, for example, 0.1N HCl. It is well accepted that 0.1N HCl
(simulated gastric fluid) serves as a proxy for the acidity of
stomach contents. Accordingly, and without being bound by theory,
it is believed that the compositions described herein can
self-micellize in situ in the stomach or small intestine. In
certain embodiments, the compositions described herein more
efficiently and effectively deliver omega-3 fatty acid esters
through the intestinal tract when administered with or without
food.
[0291] In addition to forming micelles in situ, in other
embodiments, compositions comprising micelles are provided, wherein
the micelles are formed by the addition of an aqueous medium to a
composition of any one of the embodiments provided herein prior to
administration of said composition to a subject in need of
treatment. Alternatively, micelles can also be formed when the
compositions are added to an aqueous medium. In certain
embodiments, the micelles have a diameter of up to 10 .mu.m. In
other embodiments, substantially all of the micelles have an
average diameter of from 1 .mu.m to 10 .mu.m. In certain
embodiments, the micelles have an average diameter of, for example,
1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 .mu.m. In certain embodiments, said
micelles are stable at ambient temperature and in certain other
embodiments, said micelles are stable at about normal mammalian
body temperatures.
[0292] The surface active agents comprising the compositions
suitable for self-micellization as described herein generally have
an HLB from, for example, 12 to 18. In certain embodiments, said
surface active agents have an HLB from, for example, 12.0 to 14.0.
In certain embodiments, said surface active agents have an HLB
from, for example, 13.0 to 14.0. In certain embodiments, said
surface active agents have an HLB from, for example, 13.5 to 13.8.
The total HLB of all the surface active agents or surfactants used
in the composition is generally from, for example, 12 to 18. In
some embodiments, the total HLB of all surface active agents used
in the composition is generally from, for example, 12 to 15. In
some embodiments, the total HLB of all surface active agents or
surfactants used in the composition is generally from, for example,
13 to 15.
[0293] In certain embodiments, the at least one surface active
agent or surfactant has an HLB of at least, for example, 8.0. In
some embodiments, said surface active agent(s) or surfactant(s)
have a combined HLB in the range of from, for example, 13 to 15. As
the HLB value of the surface active agent(s) or surfactant(s)
increases, the amount of surface active agent or surfactant needs
to be decreased.
[0294] The compositions described herein can further comprise at
least one antioxidant. The antioxidant(s) suitable for use in the
instant omega-3 fatty acid compositions, include, but are not
limited to tocopherols and/or tocotrienols and can be present from,
for example, 0.01% to 5% (wt/wt) of the composition. In certain
such embodiments, the tocopherols and/or tocotrienols can be
present at, for example, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%,
0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%,
4.5% or 5% by weight of the compositions. In certain such
embodiments, the antioxidant is .alpha.-tocopherol present at, for
example, 2% by weight of the composition.
[0295] In certain embodiments, the composition further comprises a
terpene. In certain embodiments, the terpene is d-limonene. In one
embodiment, the terpene is a cyclic terpene. In one embodiment, the
terpene is d-limonene ((+)-limonene), which is the (R)-enantiomer.
In one embodiment, the terpene is L-limonene, which is the
(S)-enantiomer. In one embodiment, the terpene is racemic limonene,
known as dipentene. In another embodiment, the terpene is a
terpenoid. In another embodiment, the terpene or terpenes are
derived from a natural oil (e.g., a citrus oil such as orange oil).
Other terpenes are contemplated, such as monoterpenes (e.g.,
terpinenes, terpinolenes, phellandrenes, or menthol), having
structures that are similar to d-limonene. In certain embodiments,
the compositions further comprise substantially pure d-limonene
from, for example, 0.1% to 5% by weight of the composition. In
certain other embodiments, the compositions further comprise
natural orange oil from, for example, 0.1% to 5% by weight of the
composition. Compositions comprising d-limonene or orange oil can
aid in the elimination and/or minimization of side effects from the
oral administration of the instant omega-3 fatty acid compositions.
Such side effects include regurgitation, frequency of belching,
gastroesophageal reflux disease (GERD), bloating, increased
intestinal gas, fish taste, fishy breath, fish smell, nausea,
diarrhea, or combinations thereof.
Methods for Treating Age-Related Macular Degeneration
[0296] Methods are provided of treating AMD in a subject in need of
treatment, which method comprises administering to said subject a
therapeutically effective amount of a composition of any one of the
embodiments provided herein, or a pre-formed micelles according to
the respective embodiments described herein. The compositions
described herein can be administered in conjunction with other
medications and/or nutritional supplements prescribed for treating
AMD. In certain embodiments, non-limiting examples of non-omega-3
fatty acid nutritional supplements include, for example, a
combination of vitamin C, vitamin E, beta-carotene, zinc, copper,
magnesium, manganese, calcium, vitamin A, vitamin D, vitamin K2,
lutein and zeaxanthin. Non-limiting examples of non-omega-3 fatty
acid active agents include, for example, verteporfin (VISUDYNE);
the antioxidant carotenoids crocin and crocetin, as found in, for
example, Saffron (Crocus sativus), and inhibitors of angiogenesis,
such as for example, BEVACIZUMAB (AVASTIN.RTM.), RANIBIZUMAB
(LUCENTIS.RTM.), PEGAPTANIB (MACUGEN.RTM.) AND AFLIBERCEPT
(EYLEA.RTM.).
[0297] The compositions described herein can also be administered
for treating Juvenile Macular Degeneration, which includes
Stargardt's disease, Best disease, and juvenile retinoschisis by
administering the compositions described herein to a human
juvenile.
[0298] In certain embodiments, the amount of the non-omega-3 fatty
acid active agents administered in conjunction with the
compositions described herein can be reduced to avoid or minimize
any side effects resulting from the non-omega-3 fatty acid active
agents when such agents are administered alone at higher
concentrations.
[0299] In at least one embodiment, a method is provided for
treating AMD by administering, to a patient in need thereof, a
therapeutically effective amount of a composition comprising
omega-3 DPA fatty acid (either in the triglyceride, ester or free
fatty acid ester form) and at least one surface active agent in an
amount and a combination effective to cause said compositions to
spontaneously self-micellize when in contact with an aqueous
medium, thereby forming a plurality of stable micelles having a
particle size within a range of about 1 .mu.m to about 10
.mu.m.
Methods for Treating Fat Malabsorption Syndromes or Disorders
[0300] Methods are provided for treating one or more fat
malabsorption syndrome or disorder in a subject in need of
treatment, which method comprises administering to said subject a
therapeutically effective amount of a composition of any one of the
embodiments provided herein, or a pre-formed micelle mixture of any
one of the embodiments provided herein.
[0301] Accordingly, in certain embodiments, the fat malabsorption
syndrome can be the result of a disorder in the intestinal
processes of digestion and/or transport of nutrients across the
intestinal mucosa into the systemic circulation. The fat
malabsorption syndrome can be either a congenital abnormality in
the digestive or absorptive processes or a secondarily acquired
disorder resulting from a disorder of the intestinal processes of
digestion and/or transport of nutrients across the intestinal
mucosa into the systemic circulation.
[0302] Exocrine pancreatic insufficiency can also result in a fat
malabsorption syndrome or disorder. Exocrine pancreatic
insufficiency can be the result of pancreatitis, pancreatic cancer,
pancreatic resection, cystic fibrosis, Shwachman-Diamond syndrome,
Johnson-Blizzard syndrome, and Pearson syndrome. Accordingly, in
certain embodiments, the compositions described herein can be used
for the treatment of fat malabsorption syndrome resulting from
exocrine pancreatic insufficiency.
[0303] Obstructive biliary or cholestatic liver disease or
extensive intestinal mucosal disease, such as that which occurs in
celiac disease can result in fat malabsorption syndromes. Thus, in
certain embodiments, the compositions described herein can be
administered to a human in need of such administration for the
treatment of fat malabsorption resulting from obstructive biliary
or cholestatic liver disease or extensive intestinal mucosal
disease.
[0304] Fat malabsorption syndromes can be the result of functional
or anatomic loss of extensive segments of small intestine leading
to a severe decrease in intestinal absorptive capacity, often
referred to as Short-Bowel Syndrome (SBS), which may be congenital
or acquired as a result of surgery due to Crohn's Disease or
necrotizing enterocolitis in infants or neonates. In certain
embodiments, the administration of the compositions described
herein to a human, particularly a neonate, in need of such
administration, can treat fat malabsorption syndromes resulting
from SBS or Crohn's Disease.
[0305] In order to increase the absorption of dietary fats,
including docosahexaenoic acid, through enteral administration, a
combination of fatty acids and surface active agents, specifically
poloxamers and polysorbates, can be combined to form
self-micellizing micelles. Attempts to create artificial micelles
have generally resulted in artificial micelles that are fragile and
break easily, making it difficult for the particles to deliver
adequate amounts of free fatty acids and monoglycerides to the
intestinal lining for absorption. For this reason, creating
micelles mimicking necessary characteristics of natural human
micelles is important, especially having a size of about five
microns in diameter to carry its cargo of dietary fats (or other
lipophilic materials) to and through the intestinal cell wall is a
novel means of creating bioavailable dietary fats for the treatment
of malabsorption.
[0306] In one exemplary embodiment of the present invention, a
novel formulation was utilized containing docosahexaenoic acid in
combination with a blend of surface active agents including
polysorbates and poloxamers, and a study was carried out in a
porcine model of SBS. The methodology and results of this
experiment are set forth in Example 3.
[0307] It was concluded that enteral administration of a novel DHA
preparation that forms micelles independent of bile salts results
in increased fatty acid absorption, weight gain and improved
intestinal morphologic adaptation in a neonatal porcine model of
SBS.
[0308] In certain embodiments, administration of the compositions
described herein can be administered in conjunction with a
non-omega-3 fatty acid active agent for the treatment of SBS.
Non-limiting examples of such non-omega-3 fatty acid active agents
can include L-GLUTAMINE (NUTRESTORE.RTM.), RECOMBINANT SOMATOTROPIN
(ZORBTIVE.RTM.), AND TEDUGLUTIDE (GATTEX.RTM.). Other such
non-omega-3 fatty acid active agents may include conjugated bile
acids or opium tincture. In certain embodiments, the amount of the
non-omega-3 fatty acid active agent(s) administered in conjunction
with the compositions described herein can be reduced to avoid or
minimize any side effects resulting from the non-omega-3 fatty acid
active agents when such agents are administered alone at higher
concentrations.
Methods for Treating NAFLD and/or NASH
[0309] Methods are provided of treating NAFLD and/or NASH in a
subject in need of treatment, which method comprises administering
to said subject a therapeutically effective amount of a composition
of any one of the embodiments provided herein, or pre-formed
micelles according to the respective embodiments described
herein.
[0310] The compositions described herein can be administered
concomitantly with other non-omega-3 fatty acid medications
prescribed for treating NAFLD and/or NASH. Non-limiting examples of
such non-omega-3 fatty medications can include, lipid lowering or
cholesterol lowering agents selected from the group consisting of
cholesterol absorption inhibitors, bile acid sequestrants/resins,
statins, niacin and derivatives, MTP inhibitors, fibrates and CETP
inhibitors, insulin sensitizers, hypolipidemics, anti-inflammatory
medications, and thiazolidinediones. In certain embodiments, the
amount of the non-omega-3 fatty acid active agent(s) administered
in conjunction with the compositions described herein can be
reduced to avoid or minimize any side effects resulting from the
non-omega-3 fatty acid active agents when such agents are
administered alone at higher concentrations.
[0311] In certain embodiments, the compositions, or a plurality of
micelles pre-formed therefrom, described herein, provide for a
method of treating a Non-Alcoholic Fatty acid Liver Disease (NAFLD)
comprising the step of administering to a subject in need of such
administration the compositions or a plurality of micelles
pre-formed therefrom described herein.
[0312] In certain embodiments, the compositions, or a plurality of
micelles pre-formed therefrom, described herein, provide for a
method of treating Non-Alcoholic Steatohepatitis (NASH) comprising
the step of administering to a subject in need of such
administration the compositions or a plurality of micelles
pre-formed therefrom described herein.
[0313] In certain embodiments, the compositions, or a plurality of
micelles pre-formed therefrom, described herein, provide for a
reduction in body weight (or weight loss) in a subject diagnosed
with NAFLD and/or NASH.
[0314] In certain embodiments, the compositions, or a plurality of
micelles pre-formed therefrom, described herein, provide for a
method of reducing liver weight in a subject diagnosed with
NASH.
[0315] In certain embodiments, the compositions, or a plurality of
micelles pre-formed therefrom, described herein, provide for a
method of reducing whole blood glucose in a subject diagnosed with
NASH.
[0316] In certain embodiments, the compositions, or a plurality of
micelles pre-formed therefrom, described herein, provide for a
method of reducing liver triglyceride levels in a subject diagnosed
with NAFLD and/or NASH.
[0317] In certain embodiments, the compositions, or a plurality of
micelles pre-formed therefrom, described herein, provide for a
method of reducing development of liver fibrosis in a subject
diagnosed with NAFLD and/or NASH.
[0318] A nine-week preclinical study of certain embodiments of the
present invention was conducted using a mouse model which
progresses from NAFLD to NASH between 5 to 9 weeks of age. Within
the disease progression of this animal model, the liver
triglycerides reach a point around week 6 where the inflammation
has progressed substantially over week 5. The study contained three
groups, a control group given a placebo from age five to nine
weeks, and two trial groups, a first trial group administered a
composition disclosed herein for 4 weeks, from five to nine weeks
of age, and a second trial group administered the same said
composition disclosed herein for 3 weeks, from six to nine weeks of
age.
[0319] The study showed that animals for which treatment was
started later, the 3 week treatment group, had significantly
reduced liver triglyceride contents compared with the control
group. Although the liver triglycerides tended to decrease in both
treatment groups, the 4 week treated group (from five to nine
weeks) showed a non-significant change (at 0.05) and the group
treated for 3 weeks (from six to nine weeks) showed a statistically
significant decrease, as illustrated in FIG. 3. This 3 week
treatment group also showed a tendency for reduced whole blood
glucose levels compared with the control group. The significant
decrease in the liver triglycerides in the more progressed group
demonstrate that certain embodiments of the present invention can
improve lipid and glucose metabolism in the liver. These results
also demonstrate that omega-3 fatty acids can improve lipid and
glucose metabolism in the liver.
[0320] The effect of the treatment was not merely through an
induction of weight loss by the animals. In fact, the study found
that body weight gradually increased during the treatment period in
all groups and that there was no difference in mean body weight
between groups. Although the end liver weight of the subjects in
the treatment groups was lower than that of the subjects in the
control group, the difference was not statistically significant. In
addition, both treatment groups trended to reduce alpha-SMA (a
marker of myofibroblast associated with the development of liver
fibrosis) mRNA expressions in the liver, a key indicator of the
treatment's demonstrated ability to delay the progression to NASH.
Without being held to any one theory, we believe the study
demonstrates the ability of at least one composition disclosed
herein to deliver polyunsaturated EPA and DHA fatty acids to
mammals with NAFLD for incorporation into the cell membrane of
erythrocytes, wherein said delivery can reduce the triglyceride
level in the liver and thereby slow the disease's progression to
NASH. The study may also indicate that administration of
compositions disclosed herein might provide a larger medicinal
response when provided later in the disease progression from NAFLD
to NASH.
Methods for Treating Neurodegenerative Diseases
[0321] Methods are provided of treating neurodegenerative disease
in a subject in need of treatment, which method comprises
administering to said subject a therapeutically effective amount of
a composition of any one of the embodiments provided herein, or
pre-formed micelles according to the respective embodiments
described herein. Non-limiting examples of neurodegenerative
diseases that can be treated by the compositions described herein
include PD, AD, MS, Epilepsy, and ALS.
[0322] The compositions described herein can be administered
concomitantly with other non-omega-3 fatty acid medications
prescribed for treating neurodegenerative diseases. Non-limiting
examples of medications can include medications generally approved
by a health regulatory body and prescribed by physicians for a
particular neurodegenerative disease. In certain embodiments, the
amount of the non-omega-3 fatty acid active agent(s) administered
in conjunction with the compositions described herein can be
reduced to avoid or minimize any side effects resulting from the
non-omega-3 fatty acid active agents when such agents are
administered alone at higher concentrations.
Methods for Treating Primary Sclerosing Cholangitis
[0323] Methods are provided of treating PSC and or NSC in a subject
in need of treatment, which method comprises administering to said
subject, typically a human adult, infant, child or adolescent, a
therapeutically effective amount of a composition of any one of the
embodiments provided herein, or pre-formed micelles according to
the respective embodiments described herein.
[0324] PSC, which is a chronic cholestatic liver disease, can
result in fat malabsorption syndromes. Thus, in certain
embodiments, the compositions described herein can be administered
to a human in need of such administration for the treatment of fat
malabsorption resulting from PSC.
[0325] In certain embodiments, the compositions described herein
can be administered in conjunction with non-omega-3 fatty acid
active agents. Examples of such active agents can, upon approval by
a respective regulatory agency, include
6-alpha-ethylchenodeoxycholic and/or
(4R,5R)-1-[[4-[[4-[3,3-Dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hyd-
roxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]methyl]phenyl]methyl]-4-aza-1-
-azoniabicyclo[2.2.2]octane chloride.
[0326] In certain embodiments, the amount of the non-omega-3 fatty
acid active agents administered in conjunction with the
compositions described herein can be reduced to avoid or minimize
any side effects resulting from the non-omega-3 fatty acid active
agents when such agents are administered alone at higher
concentrations.
Methods for Treating Sickle Cell Disease
[0327] Methods are provided of treating sickle cell disease in a
subject in need of treatment, which method comprises administering
to said subject a therapeutically effective amount of a composition
of any one of the embodiments provided herein, or pre-formed
micelles according to the respective embodiments described
herein.
[0328] The compositions described herein can be administered
concomitantly with other non-omega-3 fatty acid active agents and
non-omega-3 nutritional supplements prescribed for treating sickle
cell disease. Non-limiting examples of non-omega-3 active agents
can include pain medications, antibiotics, hydroxyurea,
anti-inflammatory medications, and aspirin, particularly a low-dose
(70-81 mg) aspirin dosage form; while non-omega-3 nutritional
supplements may include one or more of folic acid, magnesium,
manganese, zinc, calcium, vitamin A, vitamin E, vitamin D, and
vitamin K2. In certain embodiments, the amount of the non-omega-3
fatty acid active agent(s) administered in conjunction with the
compositions described herein can be reduced to avoid or minimize
any side effects resulting from the non-omega-3 fatty acid active
agents when such agents are administered alone at higher
concentrations.
[0329] While not wishing to be bound to any particular theories, it
is believed that compositions comprising omega-3 DPA fatty acid
(either in the triglyceride, ester or free fatty acid ester form)
and at least one surface active agent in an amount and a
combination effective to cause said compositions to self-micellize
when in contact with an aqueous medium can be used to treat sickle
cell disease symptoms, such as vaso-occlusive events, sequestration
crisis, avascular necrosis, stroke, and sickle cell anemia, among
other known maladies, by decreasing the aggregation of platelets,
red blood cells, and/or white blood cells.
[0330] Nutritional Factors Associated with SCD Pathophysiological
and Clinical Outcomes
[0331] (A) Long Chain Polyunsaturated Fatty Acids (LCPUFA)
[0332] Until recently, cell membrane lipids (fatty acids) were
regarded as biologically inert compounds that provide a
permeability barrier between interior and exterior compartments
within and between cells. It is now recognized that lipids,
independently and in concert with proteins, are central to the
regulation of cell and sub-cellular functions including signaling
and gene expression. A third of all proteins are in membranes.
These proteins, which are responsible for signaling, transport and
oxidative protection and other vital cellular processes, are
dependent on membrane lipids for their optimal function.
Experimental and human studies demonstrate that abnormal
composition, perturbation or deviation from normality of lipids
leads to cell dysfunction.
[0333] Studies have shown that variation in dietary composition of
saturated and monosaturated fatty acids cause very little change in
percent saturation of membrane lipids. In contrast, membrane lipids
are very responsive to variation to dietary composition of LCPUFA.
The greatest sensitivity to changes in dietary fatty acids was for
n-3 PUFA and n-6/n-3 fatty acids ratio. A 300% increase in red
blood DHA and EPA composition has been observed in subjects
supplemented with 1296 mg EPA and 864 mg DHA. Similarly, a 10 fold
increase in EPA concentration of mononuclear cell membrane
phospholipids has been observed four weeks after supplementation.
Studies using multiple doses of fish oil show that the
incorporation of these fatty acids in immune cells occurs in a
manner that is highly correlated with the amount of the fatty acid
consumed. Typically the increase in content of n-3 PUFAs occurs at
the expense of n-6 PUFAs, especially arachidonic acid.
[0334] Unlike n-6 fatty acids, n-3 fatty acids do not occur in
large amounts in plants food and western human diet. Moreover, the
alpha-linolenic acid (ALA) conversion process to EPA and DHA in
humans is not efficient as only 5-10% are converted to EPA, and a
mere 2-5% to DHA. N-3 fatty acids are abundantly present in fish
and shellfish. In fact, fish-oil supplements typically contain
30-50% of n-3 FAs. The current adequate intake (AI) for ALA issued
by the Institute of Medicine of the National Academies, USA is 1.6
g/day for men 19-47 years and 1.1 g/day for women 19-47 years. The
acceptable macronutrient distribution range (AMDR) for ALA is
0.6-1.2% of energy. The lower boundary of the range meets the AI
for ALA. Approximately 10% of the AMDR for ALA can be consumed as
EPA and/or DHA. The dietary guidelines (2005) also note that
consumption of approximately two servings of fish per week
(approximately 224 g total) may reduce the risk of mortality from
coronary heart disease. However, since the physiological potency of
EPA and DHA is greater than ALA and there has been a substantive
increase in the evidence base about the health benefits of long
chain omega-3 fatty acids.
[0335] Perturbation of cell membrane fatty acids composition has
been observed on red blood cells, platelets and mononuclear cells
of patients with SCD. The abnormality is characterized by high
omega-6, low omega-3 and an imbalance between the two fatty acid
families. The n-3 and n-6 LCPUFA are vital structural and
functional components of cell and sub-cellular components. Studies
have shown that the balance between these two fatty acid families
influences blood cell adhesion, aggregation, blood coagulation,
cell deformability and inflammatory response. Hence, it has been
postulated that an imbalance in membrane n-6/n-3 LCPUFA is the
antecedent of the loss of membrane asymmetry, blood cell adhesion
and aggregation and vaso-occlusion in SCD. Interestingly,
experimental and clinical studies have shown that supplementing SCD
patients with omega-3 fatty acids corrects cell membranes
abnormalities and confer protection against vaso-occlusive pain
episodes, severe anaemia and oxidative stress, and improves red
blood cells flexibility in mice, modulates the eicosanoids and
resolvin production, expression of adhesive molecules, and the
overall inflammatory state. The retroconversion of DHA to satisfy
the omega-3 deficiencies at this cellular level results in a
normalization of the cell membrane and alleviation of symptoms of
the disease state. The displacement of ARA during this process
decreases systemic inflammation.
[0336] (B) Antioxidants
[0337] Patients with sickle cell disease are under oxidative
overload caused by accelerated auto-oxidation and iron
de-compartmentalization. Among the factors thought to participate
in systemic high oxidative stress (OS) associated with SCD is the
increased intravascular haemolysis and excessive levels of
cell-free haemoglobin with its catalytic action on oxidative
reaction. In addition, a chronic pro-inflammatory state is a
characteristic of SCD patients even in steady state, and reactive
oxygen species (ROS) production is a main feature of and plays an
important role in inflammation. It has also been demonstrated that
hypoxia-reoxygenation cycles in SCD are associated with increased
OS and severe reperfusion injury. On the other hand, it has been
found that the anti-oxidant capacity in RBCs of SCD patients is
highly impaired. Thus, the chronic OS state associated with SCD is
an outcome of imbalance between enhanced generation of reactive
oxygen species (ROS) and disrupted antioxidant system. It has been
postulated that the impaired antioxidant capacities could be due to
an accelerated utilization of antioxidant nutrients. There is
evidence to indicate that patients with SCD have a reduced levels
of vitamin A, C, E and beta carotene and zinc.
[0338] (C) Protein
[0339] Sickle cell disease is associated with hyper-metabolism and
a consequent shortage of substrates for normal growth and healthy
immune response. Insufficient nutrition in general is associated
with poor immune function and is consequently regarded as the most
common cause of immunodeficiency worldwide. The protein:energy
ratio is a major determinant of dietary adequacy.
[0340] Experimental studies have demonstrated that high protein
intake results in lower circulating inflammatory markers, fewer
infarcts in spleen, liver, and kidney, and lower histopathologic
scores for chronic tissue injury in liver and spleen. In addition,
high-protein fed sickle mice had less vascular leakage in the
heart, lungs, and brain and a better acute immune response and
survival rate than matched normal-protein sickle mice. These
experimental studies suggest that that the difference in
nutritional intake might be a major contributory factor that
explains partially the wide spectrum of SCD clinical manifestation
and severity. However, to the best to our knowledge, no study has
investigated the long-term effect of low or high protein and energy
intake on SCD acute and chronic complications.
[0341] Based on the previous studies on nutritional modifiers of
sickle cell disease and the FDA approved Daily Reference Values
(DRVs) and Reference Daily Intakes (RDIs) for adults and children
four or more years of age, the following dietary formula to prevent
acute and chronic complications associated with SCD:
Protein: 50 g
Vitamin A: 5,000 International Units (IU)
Vitamin C: 60 mg
Vitamin E: 30 IU
[0342] Beta carotene:
Folate: 400 .mu.g
Zinc: 15 mg
Manganese: 2 mg
Copper: 2 mg
Selenium: 70 .mu.g
[0343] Omega-3 fatty acids (DHA AND EPA): 500 mg
[0344] Kits
[0345] Packaged pharmaceutical kits comprising the compositions
described herein are contemplated. The kits comprise the
compositions described herein as unit dosage forms in a container
made of an inert material, such as for example, a glass vial, which
can be clear or colored. The glass vial can be a crimp or snap top
glass vial. Included in the kits are instructions for using the
dosage form to treat a subject having a disease or disorder
responsive to treatment by administration of the dosage forms
comprising the compositions described herein.
[0346] The packaged pharmaceutical kits provide prescribing
information, over the counter medical use information, and/or
nutritional information for the dosage form including, for example
and without limitation, to a subject or health care provider, or as
a label in a packaged pharmaceutical kit. Information included in
the kit may include, for example and without limitation, efficacy,
dosage and administration, contraindication and adverse reaction
information pertaining to the omega-3 fatty acid compositions
described herein. The dosage and administration information, for
example, can include dosing frequency recommendations as well as
administration of the compositions with or without food.
[0347] In certain embodiments the dosage forms comprising the
compositions provided herein are in the form of a liquid filled or
gel filled capsules for oral administration, which are provided
either as blister packages or in bottles together with over the
counter or prescription medical use information and/or nutritional
information.
[0348] The packaged pharmaceutical kits can comprise one or more of
the omega-3 fatty acids comprising the compositions described
herein as the only active ingredient. In other embodiments, one or
more of the compositions described herein can be packaged in
combination with one or more active agents other than a non-Omega 3
fatty acid, such as for example and without limitation, one or more
other lipid lowering or cholesterol lowering agents selected from
the group consisting of cholesterol absorption inhibitors, bile
acid sequestrants/resins, statins, niacin and derivatives, MTP
inhibitors, fibrates and CETP inhibitors,
6-alpha-ethylchenodeoxycholic and/or
(4R,5R)-1-[[4-[[4-[3,3-Dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hyd-
roxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]methyl]phenyl]methyl]-4-aza-1-
-azoniabicyclo[2.2.2]octane chloride, insulin sensitizers,
hypolipidemics, anti-inflammatory medications, and
thiazolidinediones, pain medications, antibiotics, folic acid,
hydroxyurea, and anti-inflammatory medications.
[0349] In certain embodiments such active agents can include
medications generally approved by a health regulatory body and
prescribed by physicians for a particular neurodegenerative
disease.
[0350] In other embodiments, one or more of the compositions
described herein can be packaged in combination with one or more
nutritional supplements. Non-limiting examples of supplements
include various minerals, such as calcium, magnesium, iron, and
vitamins, particularly fat-soluble vitamins, as well protein
supplements well known in the art.
[0351] Dosage Forms
[0352] Any of the omega fatty acid containing compositions provided
herein can be provided as a pharmaceutical composition to be
administered orally or parenterally, as a nutraceutical
formulation, or a dietary supplement.
[0353] In certain embodiments of the invention, compositions
comprise at least one surface active agent, at least one
fat-soluble vitamin, such as vitamin A, vitamin D, vitamin K,
vitamin K2, among others, and an omega-3 fatty acid (in
triglyceride, ester, or free fatty acid form). In certain
embodiments of the invention, at least one surface active agent is
combined with at least one fat-soluble vitamin and an omega-3 fatty
acid (in triglyceride, ester, or free fatty acid form) in relative
concentrations sufficient to form micelles when added to an aqueous
medium. In certain other embodiments of the invention, at least one
surface active agent is combined with at least one fat-soluble
vitamin, an omega-3 fatty acid (in triglyceride, ester, or free
fatty acid form), and one or more minerals, such as magnesium,
manganese, zinc, copper, selenium, and combinations thereof.
[0354] In certain embodiments of the invention, at least one
surface active agent is combined with at least one fat-soluble
vitamin, an omega-3 fatty acid (in triglyceride, ester, or free
fatty acid form), and one or more minerals, such as magnesium,
manganese, zinc, copper, selenium, and combinations thereof, such
that when added to an aqueous medium, the composition forms
micelles and wherein said micelles range in size from 1 um to 10 um
in diameter.
[0355] The pharmaceutical compositions described herein may further
include one or more pharmaceutically acceptable excipients.
Pharmaceutically acceptable excipients include, but are not limited
to, carriers, preservatives, and/or coloring agents. General
considerations in the composition and/or manufacture of
pharmaceutical compositions may be found, for example, in Remington
The Science and Practice of Pharmacy 21st ed., Lippincott Williams
& Wilkins, 2005.
[0356] In certain embodiments, the compositions described herein
can be formulated as a liquid for oral administration. Liquid
compositions include solutions, suspensions and emulsions. Examples
of liquid pharmaceutical preparations include propylene glycol
solutions and solutions containing sweeteners for oral solutions,
suspensions and emulsions. When the liquid composition comes into
contact with an aqueous medium, such as for example an aqueous
medium having an acidic environment, the composition forms
micelles.
[0357] In certain embodiments, the dosage form comprises micelles
which may be pre-formed prior to administration to a subject in
need of such administration, and wherein said micelles range in
size from 1 um to 10 um in diameter. Such pre-formed micelles are
stable at room temperature.
[0358] In other embodiments, the compositions described herein can
be formulated as a fill material for capsules, including for
example, a soft gelatin capsule. Likewise, when the contents of the
soft gelatin capsule comes into contact with an aqueous medium, the
composition forms micelles upon disintegration of the capsule.
[0359] A capsule may be prepared, e.g., by placing the compositions
described above inside a capsule shell. A capsule is a dosage form
administered in a special container or enclosure containing an
active agent. In some embodiments the compositions described herein
can be filled into soft capsules. A capsule shell may be made of
methylcellulose, hydroxypropylmethyl cellulose, polyvinyl alcohols,
or denatured gelatins or starch or other material. Hard shell
capsules are typically made of blends of relatively high gel
strength bone and pork skin gelatins. In some embodiments the unit
dosage form is a gel capsule. In some embodiments the capsule shell
is a glycerin capsule shell, for example product no. GSU0051
manufactured by SwissCaps and which meets USP 25 requirements
(SwissCaps, USA 14193 SW 119th Ave., Miami/Fla., U.S. 33186). In
other embodiments the capsule is a bovine gelatin shell, for
example SwissCaps product no. GSU0708. Other suitable capsule shell
materials include polyethylene, polypropylene,
poly(methylmethacrylate), polyvinylchloride, polystyrene,
polyurethanes, polytetrafluoroethylene, nylons, polyformaldehydes,
polyesters, cellulose acetate, and nitrocellulose. The capsule
shell itself may contain small amounts of dyes, opaquing agents,
plasticizers, and preservatives. Conventional methods for preparing
other solid dosage forms, for example, capsules, suppositories, and
the like are also well known. Gelatin capsule shells may be made
also be made of tapioca, grass, vegetable derived or fish derived
gelatin. For example K-CAPS (Capsuline, Inc. Pompano Beach, Fla.)
is a certified Kosher soft capsule shell of vegetable origin. Other
vegetarian derived gelatin capsules may, be made of vegetable
derived hydroxypropylmethyl cellulose (HPMC). Capsules shells may
also contain Modified Maize Starch, Glycerol, and Carrageenan as a
gelling agent.
[0360] In other embodiments the capsule has a shell comprising the
material of the rate-limiting membrane, including coating
materials, and filled with the compositions described herein.
Capsule shells may be made of a porous or a pH-sensitive polymer
made by a thermal forming process. In certain embodiments the
capsule shell in the form of an asymmetric membrane; i.e., a
membrane that has a thin skin on one surface and most of whose
thickness is constituted of a highly permeable porous material.
[0361] Yet another useful capsule, a "swelling plug device", can be
used. The compositions described herein can be incorporated into a
non-dissolving capsule-half of the device which is sealed at one
end by a hydrogel plug. This hydrogel plug swells in an aqueous
environment, and, after swelling for a predetermined time, exits
the capsule thus opening a port through which the active agent can
leave the capsule and be delivered to the aqueous environment.
Preferred hydrogel-plugged capsules are those which exhibit
substantially no release of active agent from the dosage form until
the dosage form has exited the stomach and has resided in the small
intestine for about 15 minutes or more, preferably about 30 minutes
or more, thus assuring that minimal omega-3 fatty acid (either in
the triglyceride, ester, or free fatty acid form) is released in
the stomach or the small intestine. Hydrogel-plugged capsules of
this type have been described in patent application WO90/19168.
[0362] The dosage forms may contain a plasticizer, such as for
example, in a capsule shell. Suitable plasticizers include, e.g.,
polyethylene glycols such as PEG 300, PEG 400, PEG 600, PEG 1450,
PEG 3350, and PEG 800, stearic acid, propylene glycol, oleic acid,
triethyl cellulose, triacetin, glycerin, sorbitol, sorbitan or
combinations thereof.
[0363] In additional embodiments, the compositions can be
formulated as a liquid for parenteral administration, particularly
for intravenous administration, added to infant formula, or
injected into an intravenous bag comprising a saline or other
pharmaceutically acceptable solution or a nutritional
supplement.
[0364] The compositions described herein can be formulated as one
or more dosage units. In some embodiments, it can be advantageous
to formulate oral compositions in dosage unit form for ease of
administration and uniformity of dosage. Dosage unit forms
described in some embodiments can refer to physically discrete
units suited as unitary dosages for the subject to be treated; each
unit containing a predetermined quantity of active composition
calculated to produce the desired therapeutic effect in association
with the suitable pharmaceutical carrier.
[0365] In certain embodiments, the dosage form may optionally
contain a flavorant such as orange oil, substantially pure
d-limonene, and an antioxidant such as tocopherol, tocotrienol,
ascorbyl palmitate or a combination of antioxidants.
[0366] Functional Foods
[0367] Certain embodiments of the present invention provide for
functional foods comprising compositions which contain at least one
surface active agent and at least one omega-3 fatty acid (in
triglyceride, ester, or free fatty acid form) in relative
concentrations sufficient to form micelles when added to an aqueous
medium, and wherein said micelles range in size from 1 um to 10 um
in diameter. These functional foods may further contain one or more
of (1) at least one fat-soluble vitamin, such as vitamin A, vitamin
E, vitamin D, vitamin K, vitamin K2, among others, (2) one or more
minerals, such as magnesium, manganese, zinc, copper, selenium, and
combinations thereof, and (3) an omega-6, for example arachidonic
acid (ARA), linoleic acid (LA), gamma-linolenic acid (GLA) or the
omega-6 isomer of DPA.
[0368] In certain embodiments, the compositions described herein
comprise micelles pre-formed prior to administration to a subject
in need of such administration. Such pre-formed micelles are stable
at room temperature. Accordingly, either such pre-formed micelles
or said pre-micellized compositions in combination with a
foodstuff, are provided, and can then be consumed as part of a
healthy diet for enriching a subject's omega-3 fatty acid levels or
as a dietary treatment in addition to the oral/parenteral
administration of the compositions described herein as prescribed
by a health professional.
[0369] In certain embodiments, the functional food is in the form
of edible or drinkable compositions, e.g., foodstuffs such as
chewable or edible bars, confectionary products (e.g., chocolate
bars), cookies, juice drinks, baked or simulated baked goods (e.g.,
brownies), biscuits, lozenges or chewing gum. Examples of chewable
or edible bars include chocolate bars or energy bars. Such
functional foods can be particularly useful to people participating
in sports or other forms of exercise.
[0370] In certain embodiments, the functional foods may also be in
the form of, for example, butter, margarine, bread, cake, milk
shakes, ice cream, yogurt and other fermented milk product.
[0371] In certain embodiments, the functional food can also be in
the form of a liquid to be sprayed on meats, salads or other
foods.
[0372] Other forms of the functional foods can be baby food,
breakfast cereals, such as for example, grain flakes, muesli, bran,
oatmeal.
[0373] When the functional food product is in a drinkable form, the
compositions described herein can be added directly to the drink,
such as for example plain milk, flavored milk, fermented milk
products or juices, or infant formula. The compositions will form
micelles comprising the omega-3 fatty acids in the drinkable
product.
[0374] When the functional food is in the form of a solid edible
product, the compositions described herein can be first added
directly to a functional food or to an aqueous medium, wherein the
composition will form micelles as described herein. The aqueous
medium comprising the micelles can subsequently be either sprayed
onto the solid edible product or mixed into the ingredients when
manufacturing the edible product.
[0375] In certain embodiments particularly for the treatment of
sickle cell disease, at least one surface active agent and one or
more omega-3 fatty acids (in triglyceride, ester, or free fatty
acid form, e.g. DHA alone or DHA and EPA: 500 mg), are further
combined with Protein: 50 g, Vitamin A: 5,000 International Units
(IU); Vitamin C: 60 mg, Vitamin E: 30 IU, Beta carotene, Folate:
400 .mu.g, Zinc: 15 mg, Manganese: 2 mg, Copper: 2 mg, and
Selenium: 70 .mu.g.
[0376] In certain embodiments of the invention, at least one
surface active agent is combined with at least one fat-soluble
vitamin and an omega-3 fatty acid (in triglyceride, ester, or free
fatty acid form) in relative concentrations sufficient to form
micelles when added to an aqueous medium. In certain other
embodiments of the invention, at least one surface active agent is
combined with at least one fat-soluble vitamin, an omega-3 fatty
acid (in triglyceride, ester, or free fatty acid form), and one or
more minerals, such as magnesium, manganese, zinc, copper,
selenium, and combinations thereof.
EXAMPLES
Macular Degeneration
Example 1
[0377] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 1:
TABLE-US-00001 TABLE 1 COMPOSITION INGREDIENT Amount (mg) % (wt/wt)
DHA Omega-3 fatty acid Ethyl 754.3 68.57 Ester*.dagger. Polysorbate
80 337.9 30.72 Poloxamer 237 (Pluronic .RTM. F87) 7.8 0.71 TOTAL
1100 100 *The omega-3 oil may contain ~2% .alpha.-tocopherol as an
antioxidant. .dagger.DHA comprises at least 90% of the DHA oil
(678.9 mg). The majority of the remaining no more than 10% is EPA
(~5%, or up to 37.7 mg)
Example 2
[0378] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 2:
TABLE-US-00002 TABLE 2 COMPOSITION INGREDIENT Amount (mg) % (wt/wt)
EPA/DHA Omega-3 fatty acid 754.3 65.59 Ethyl Ester*.dagger.
Gamma-tocotrienol 50.0 4.35 Polysorbate 80 337.9 29.38 Poloxamer
237 (Pluronic .RTM. F87) 7.8 0.68 TOTAL 1150 100 *The Omega-3 oil
may contain ~2% .alpha.-tocopherol as an antioxidant. .dagger.EPA
comprises 383 mg and 170 mg DHA.
Malabsorption Syndrome
Example 3
[0379] A study was conducted to evaluate, in an established porcine
model of SBS, the systemic absorption and intestinal adaptation
capacity of an ethyl ester form of DHA in combination with a
surface active agent composition effective to spontaneously form
micelles with said fatty acids upon contact with an aqueous media.
In certain embodiments, the self-micellizing compositions may
further contain at least one additional omega-3 fatty acid (either
in the triglyceride, ester, or free fatty acid form), for example
EPA or other known omega-3 fatty acids as disclosed herein. In
certain embodiments, the self-micellizing compositions may further
contain an anti-oxidant, such as tocopherol. In certain
embodiments, the self-micellizing compositions may further contain
an omega-6 fatty acid (either in the triglyceride, ester, or free
fatty acid form), for example arachidonic acid (ARA), linoleic acid
(LA), gamma-linolenic acid (GLA) or the omega-6 isomer of DPA, also
known as osbond acid. This composition allows for micelle formation
independent of the presence of bile salts as compared to a standard
triglyceride form of DHA without a surface active agent
(Control).
[0380] Materials and Methods
[0381] Animals and Surgical Induction of Short Bowel Syndrome
[0382] Newborn, crossbred
(Hampshire.times.Landrace.times.Duroc.times.Yorkshire) piglets
(n=10), were weighed, placed in cages in a heated room
(.about.30.degree. C.), and were fed with a commonly available sow
milk replacement formula. After an overnight fast, piglets
underwent surgery for the placement of jugular and intragastric
catheters for postoperative parenteral and enteral nutrition,
respectively, and small bowel resection surgery. Following a 4-cm
midline incision an 80% mid-jejunoileal resection was performed.
The total amount of bowel resection was approximated using the
following equation: total intestinal length=280 cm.times.body
weight{circumflex over ( )}0.60. A 20% remnant (.about.60 cm total
per kg BW) remained and was represented equally between proximal
jejunum and ileum, representing .about.30 cm each remnant segment
per kg bodyweight. .about.45 cm of jejunum distal to the ligament
of Treitz and .about.45 cm of ileum proximal to the ileocecal
junction were measured using sterile silk ribbon placed along the
antimesentric border of the gently stretched small intestine.
Intestine not included in the measurement was removed via
cauterization. Bowel continuity was restored using an end-to-end
jejunoileal anastomosis.
Postoperative Nutrition
[0383] Parenteral nutrition formula which does not contain DHA was
provided proportionately to body weight to all piglets within 3 h
after the operation. Continuous enteral feeding with a commonly
available sow milk replacer formula, also lacking DHA, was slowly
introduced 24 h after intestinal resection at 3 mL/kg*h via the
intragastric catheter. The volume of enteral feedings was then
advanced every 24 h by 1 ml/kg*h until a final enteral volume of 5
mL/kg*h was achieved on postoperative day 3 which represents
approximately 50% of full enteral feeding volume and nutrient
intake.
Provision of Oral Treatment & Control Groups
[0384] After the surgical procedure, piglets were randomly
allocated into two groups (N=5 per group), control and
SBS-treatment group. The control group received a standard DHA
ethyl ester preparation of substantially pure DHA ethyl ester
(>90% DHA ethyl ester). The SBS-treatment group received an
equivalent amount of DHA as the control group, based on animal
weight as described herein, of a preferred embodiment of the
present invention comprising, about 683.9 mg/g of substantially
pure DHA ethyl esters (>90% DHA ethyl ester), 2.7 mg/g of
alpha-tocopherol, 306.3 g/mg of polysorbate 80, and 7.1 mg/g of
poloxamer 237.
[0385] SBS-treatment: The control and SBS-treatment formulations,
respectively, mixed in the enteral feeding at the respective dose
and infused via the intragastric catheter at a dose of 1 g/kg*day
for a period of 4 days after which time the animals were
sacrificed.
[0386] Growth Data and Sample Collection
[0387] Growth
[0388] Piglets were weighed every other day. In addition to
calculating a change in weight by subtracting the starting weight
at the time of randomization from the final weight, percent weight
change and growth velocity (g/kg*d) were also calculated. Percent
weight change was calculated by: [((final weight-starting
weight)/starting weight).times.100]. Growth velocity was calculated
by: [(weight change/starting weight)/4 days of treatment].
TABLE-US-00003 TABLE 3 Cohort characteristics and weight parameters
(median .+-. IQR) Control (n = 5) Treated (n = 5) p Male, n (%)
4(80) 3(60) 0.51 Weight at start, g 2490 .+-. 114 2474 .+-. 325
0.75 Weight at finish, g 2594 .+-. 170 3170 .+-. 800 0.35 Change in
weight, g 132 .+-. 278 696 .+-. 475 0.08 Growth velocity, g/kg/d
13.5 .+-. 30.1 69.9 .+-. 41.5 0.08 Weight change, % 5.4 .+-. 12.1
28.0 .+-. 16.6 0.08
[0389] Longitudinal Plasma Fatty Acid Levels
[0390] Blood samples for systemic fatty acid profiles were obtained
on the day of surgery and then twice daily until sacrifice on
fourth postoperative day. Plasma fatty acids were isolated and
measured.
[0391] Intestinal Samples and Morphometry Measurements
[0392] Samples of intestinal tissue resected at the time of surgery
and remnants at sacrifice were collected from proximal jejunum
(proximal to anastomosis) and distal ileum (distal to anastomosis)
and morphological measurements of the crypts and villi were
measured. Only intact villi and crypts were measured. All
measurements were made by the same, single, blinded observer.
Statistical Analysis
[0393] The primary outcome measure was a change in plasma DHA level
in the SBS-treated group versus control group. Given the small
number of piglets per group, appropriate small group statistical
methods were used to evaluate differences in outcome variables.
Variable summary statistics are expressed as
median.+-.interquartile range (IQR). The Wilcoxon Rank-Sum test,
which is appropriate to compare two non-normal distributions as is
common in small sample size, was used to compare outcome measures
between groups; while the Wilcoxon signed-rank test, which is
appropriate for repeated measurements within the same animal, was
used to compare paired, before and after surgery outcome measures
within groups. Differences in growth measurements and fatty acid
levels over time between groups were evaluated using generalized
linear mixed models (GLMM) to account for repeated measures and
non-Gaussian, non-normal data.
Results
[0394] Piglet Characteristics and Weight Trends
[0395] Four of the 5 piglets were male in the control group while 3
of the 5 were male in the SBS-treatment group (Table 1). The median
starting weight was similar in both groups. At the end of the
protocol, the median difference in final weight from starting
weight was 696.+-.425 g in the SBS-treatment group compared to
132.+-.278 g in the control group. In concordance with the weight
difference the percent weight change and growth velocity was
greater in the SBS-treatment group compared to the control group.
However, due to the short duration of this model as well as the
sample size, these differences in growth parameters did not reach
statistical significance.
[0396] Small Bowel Resection and Final Organ Weights
[0397] There were no differences in the total length or weight of
resected bowel per kg weight of the piglet (Table 4). A median
total of 138.+-.34 cm per kg of small bowel was removed from
piglets in the SBS-treatment group compared to 119.+-.7 cm per kg
in the control group. Similarly, there were no differences in the
final length or weight of the jejunum and ileum segments of the
small bowel per kg weight. Finally, the liver and brain weights per
kg at sacrifice were comparable between the two groups.
TABLE-US-00004 TABLE 4 Small intestine resection and final organ
weights (median .+-. IQR) SBS- Control Treatment p Resected SI,
g/kg 29.5 .+-. 2.2 28.7 .+-. 4.9 0.92 Resected SI, cm/kg 119 .+-.
7.0 138 .+-. 3.4 0.17 Jejunum, g/kg 5.2 .+-. 1.4 5.1 .+-. 0.5 0.92
Jejunum, cm/kg 21.2 .+-. 6.7 16.5 .+-. 3.8 0.60 Ileum, g/kg 5.3
.+-. 1.2 7 .+-. 1.4 0.05 Ileum, cm/kg 17.5 .+-. 4.7 19.2 .+-. 4.4
0.33 Liver, g/kg 36.3 .+-. 2.5 37.0 .+-. 4.2 0.71 Brain, g/kg 14.0
.+-. 1.6 11.6 .+-. 2.3 0.09 IQR: Interquartile range; SI: Small
Intestine; SBS-treatment
[0398] Plasma Fatty Acid Levels
[0399] Initial plasma fatty acid levels in mol % (Time 0) of DHA,
Eicosapentaenoic acid (EPA), ARA, and Linoleic acid (LA) obtained
at the time of surgery were similar between groups (2.0.+-.0.04 vs
2.3.+-.0.4; 0.2.+-.0.06 vs 0.3.+-.0.1; 11.0.+-.0.3 vs 11.8.+-.0.5;
18.1.+-.2.3 vs 16.6.+-.1.8, respectively) (FIG. 1). Within 12 hours
(after the first enteral dose of DHA), the DHA and EPA levels were
significantly greater in the SBS-treatment group versus control
group (4.1.+-.0.3 vs 2.5 vs 0.5, p=0.009; 0.7.+-.0.3 vs
0.2.+-.0.005, p=0.009, respectively). For DHA these differences
persisted until the final measurement; while the EPA plasma level
differences persisted throughout the remaining duration of the
protocol. On the third postoperative day, plasma ARA levels began
to diverge between the groups, with lower values in the
SBS-treatment group versus the control group (7.1.+-.1.5 vs
8.7.+-.0.8, p=0.03). The difference persisted throughout the
remaining duration of the protocol. There were no differences
throughout the study protocol in plasma LA levels.
[0400] Small Bowel Morphometry
[0401] At the time of surgery (Day 0), villus height measurements
of the proximal jejunum and distal ileum were similar in both
groups (FIG. 2). In the control group, the villus height of the
proximal jejunum at the end of the experiment (Final) was not
significantly different to the values obtained at the time of
surgery; whereas in the distal ileum villus height was reduced, but
not significantly different (p=0.07). In contrast, in the
SBS-treatment group, the final villus height was increased from the
starting value in the proximal jejunum (p=04). In the distal ileum,
the SBS-treatment group demonstrated maintenance of overall villus
height and compared to the control group, a significant increase in
the final villus height (p=0.01).
[0402] Crypt depth was increased at the end of the experiment
compared to baseline values in the proximal jejunum within both
control and SBS-treatment groups (p<0.05). There were no
differences between groups in the baseline or final crypt depth
measurements. Similar to the proximal jejunum, the crypt depth of
the distal ileum was increased from baseline in the distal ileum
within the control group (p<0.05). In contrast, there were no
changes in the baseline to final crypt depth in the distal ileum
for the piglets in the SBS-treatment group. The final crypt depths
in the distal ileum were not significantly different between the
two groups.
Discussion
[0403] Piglets with surgically induced SBS who were given dietary
supplementation in the SBS-treatment group demonstrated improved
absorption of DHA, greater ileal adaptation, and a trend towards
improved growth compared to piglets given a standard preparation of
DHA. Within 12 hours, the plasma levels of both DHA and EPA in the
SBS-treatment group began to significantly diverge from the control
group. While not wishing to be bound to any particular theory, an
increase of EPA is believed to be a result of retro-conversion of
DHA to EPA as neither DHA formulation contained an EPA supplement.
The observed reduction in ARA with increasing DHA levels in the
SBS-treatment group is a known phenomenon. However, while not
wishing to be bound to any particular theory, as DHA levels
increase with DHA-only dietary supplementation, ARA levels decline
as a result of the competition for enzyme activity, the sn-2
position of phospholipids in membranes, as well as the body's
ability within the plasma compartment to maintain a constant number
of double bonds. This is not a generalized phenomenon as Linoleic
Acid levels were unchanged throughout the experimental time
period.
[0404] An increase in villus height and crypt depth is a known
observation in intestinal adaptation after significant bowel
resection in both animal models and in humans. This was observed
for both groups in the proximal jejunum and distal ileum
reinforcing the validity of this model in studying intestinal
changes in short bowel syndrome. In the proximal jejunum the
SBS-treatment group demonstrated a greater attainment in villus
height compared to baseline. In the distal ileum, the SBS-treatment
group maintained villus height while the control group lost villus
height suggesting a greater intestinal adaptive response for the
piglets receiving the SBS-treatment preparation. The piglets in the
SBS-treatment group appeared to have an accelerated adaptive
response compared to the control group possibly due to the
efficient bioavailability and intestinal absorption of DHA. This
may be a clinically relevant observation during the critical,
immediate postoperative period. Although increased jejunal crypt
depth was seen in both groups after 4 days, the lack of change in
the ileum selectively in the SBS-treatment group suggests that
increased bioavailability of DHA is exerting an intestinal trophic
effect. While not wishing to be bound to any particular theory, it
is possible that DHA is modulating intestinal cell growth and
differentiation through PPAR or other growth factors.
[0405] The example described herein provides data necessary to
support the clinical utility of a novel preparation of DHA that
allows for bile acid independent formation of micelles enhancing
absorption and biological effectiveness of DHA early in the
postoperative period.
Example 4
[0406] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 5:
TABLE-US-00005 TABLE 5 COMPOSITION INGREDIENT Amount (mg) % (wt/wt)
DHA Omega-3 fatty acid Ethyl 685.7 68.57 Ester*.dagger. Polysorbate
80 307.2 30.72 Poloxamer 237 (Pluronic .RTM. F87) 7.1 0.71 TOTAL
1000 100 *The omega-3 oil may contain ~2% .alpha.-tocopherol as an
antioxidant. .dagger.DHA comprises at least 90% of the DHA oil
(617.3 mg). The majority of the remaining no more than 10% is EPA
(~5%, or up to 34.3 mg)
[0407] During administration, sterile saline solution is added to
the vial and mixed to homogeneity to form a milky solution, which
can then be administered intravenously directly into a human in
need of such administration, such as for example a neonate, added
to infant formula, into an IV infusion bag, via a stomach feeding
tube, or through a duodenal feeding tube.
Example 5
[0408] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 6:
TABLE-US-00006 TABLE 6 COMPOSITION INGREDIENT Amount (mg) % (wt/wt)
DHA Omega-3 fatty acid Ethyl 651.4 65.14 Ester*.dagger. Arachidonic
Omega-6 Ethyl 34.3 3.43 Ester*.degree. Polysorbate 80 307.2 30.72
Poloxamer 237 (Pluronic .RTM. F87) 7.1 0.71 TOTAL 1000 100 *The
omega-3 oils may contain ~2% .alpha.-tocopherol as an antioxidant.
.dagger.DHA comprises at least 90% of the DHA oil (586.3 mg). The
majority of the remaining no more than 10% is EPA (~5%, or up to
32.6 mg) .degree.The ARA Ethyl Ester is at 80% pure.
NAFLD/NASH Examples
Example 6
[0409] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 7:
TABLE-US-00007 TABLE 7 COMPOSITION INGREDIENT Amount (mg) % (wt/wt)
DHA Omega-3 fatty acid Ethyl 754.3 68.57 Ester*.dagger. Polysorbate
80 337.9 30.72 Poloxamer 237 (Pluronic .RTM. F87) 7.8 0.71 TOTAL
1100 100 *The omega-3 oil may contain ~2% .alpha.-tocopherol as an
antioxidant. .dagger.DHA comprises at least 90% of the DHA oil
(678.9 mg). The majority of the remaining no more than 10% is EPA
(~5%, or up to 37.7 mg)
Example 7
[0410] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 8:
TABLE-US-00008 TABLE 8 COMPOSITION INGREDIENT Amount (mg) % (wt/wt)
EPA/DHA Omega-3 fatty acid Ethyl 754.3 65.59 Ester*.dagger.
Gamma-tocotrienol 50.0 4.35 Polysorbate 80 337.9 29.38 Poloxamer
237 (Pluronic .RTM. F87) 7.8 0.68 TOTAL 1150 100 *The omega-3 oil
may contain ~2% .alpha.-tocopherol as an antioxidant. .dagger.EPA
comprises 383 mg and 170 mg DHA.
Example 8
[0411] An embodiment of the present invention, referred to as SC410
herein, is a composition consisting of about: 68.39%
docosahexaenoic acid ethyl esters (DHA), 0.27% alpha-tocopherol,
30.63% polysorbate 80, and 0.71% poloxamer 237. As disclosed
herein, SC410 is preferably administered enterally to a mammal,
such as a human, and delivered within a soft gelatin capsule
although the routes of administration and delivery include other
known routes. A nine-week preclinical study of certain embodiments
of the present invention was conducted on 24 male mice using a
mouse model ("NASH mice") which progresses from NAFLD to NASH
between 5 to 6 weeks of age. Within the disease progression of this
animal model, the animals reach NASH around week 6. Eight NASH mice
were orally administered an olive oil vehicle in a volume of 2.5
mL/kg once daily from 5 to 9 weeks of age (Control Group). A second
group, the "Second Treatment Group," also interchangeably referred
to herein as NAFLD Group, composition 1-5 group, or composition 1-5
treatment, consisted of eight NASH mice which were orally
administered said olive oil vehicle supplemented with SC410 once
daily from 5 to 9 weeks of age. A third group, the "Third Treatment
Group," also interchangeably referred to herein as NASH Group,
composition 1-6 group or composition 1-6 treatment, consisted of
eight NASH mice, which were orally administered said olive oil
vehicle supplemented with SC410 once daily from 6 to 9 weeks of
age. Non-fasting blood glucose was measured in whole blood and ALT
levels were measured from plasma. Liver total lipid-extracts were
obtained from right lobes by Folch's method.
[0412] FIG. 3 shows the mean body weight on the day of sacrifice of
the mice between each of the Control, Second Treatment Group and
Third Treatment Group.
[0413] FIG. 4 shows .alpha.-SMA mRNA gene expression in mice in
each of the control, NAFLD and NASH groups treated with either
Vehicle or Composition 1 at the end of the treatment period.
[0414] FIG. 5 shows plasma alanine transaminase (ALT) levels in
mice between each of the control, NAFLD and NASH group treated with
either Vehicle or Composition at the end of the treatment
period.
[0415] FIG. 6 shows the mean liver weight on the day of sacrifice
of the mice between each of the treatment groups and the
control.
[0416] FIG. 7 shows the mean liver to body weight ratio on the day
of sacrifice of the mice between each of the treatment groups and
the control.
[0417] FIG. 8 shows whole blood glucose in mice between each of the
control, NAFLD and NASH group treated with either Vehicle or
Composition at the end of the treatment period.
[0418] FIG. 9 shows liver triglyceride levels in mice between each
of the control, NAFLD and NASH.
[0419] FIGS. 10 A and 10B show the mean individual EPA (A) and DHA
(B) total lipid concentration-time profiles (baseline-adjusted
change) after a single dose of SC401 during fed and fasting
conditions.
[0420] FIGS. 11A and 11B show the mean individual EPA (A) and DHA
(B) free fatty acid concentration-time profiles (baseline-adjusted
change) after a single dose of SC401 during fed and fasting
conditions.
[0421] FIG. 12 shows mean EPA and DHA total lipid plasma
concentration profiles (.mu.g/ml) (baseline-adjusted) after
administration of a single dose (dose adjusted) of SC401 and
Lovaza.RTM. in fasted conditions.
Note--N.S. In the Figures Stands for Non-Significant.
[0422] Body Weight
[0423] At the end of the experiment, it was determined that body
weight gradually increased during the treatment period in all
groups as would be expected in growing mice. There was no
significant difference in mean body weight between the Control
Group and the First Treatment Group during the treatment period
although mean body weight on the day of sacrifice of the First
Treatment Group tended to be decreased compared with the Control
Group. However, the mean body weight of the Second Treatment Group
was significantly lower than that of the Control Group at day
28.
[0424] .alpha.-SMA
[0425] .alpha.-SMA, a marker of myofibroblast associated with the
development of liver fibrosis, mRNA expression levels tended
non-significantly to decrease in both treatment groups as compared
with the Control Group.
[0426] ALT Measurements
[0427] Although changes in ALT were not statistically significant
between the groups, some animals within the treatment groups
exhibited a marked increase in plasma ALT levels as compared to the
control group. Since omega-3 administration has been shown in
previous studies to elevate ALT, this result would be expected and
may be an indicator of the efficacy of delivery of the omega-3 to
the liver.
[0428] Liver Weight
[0429] Additionally, there was no significant difference in mean
liver weight between the Control Group and the First Treatment
Group, whereas the mean liver weight of the Second Treatment Group
was lower than that of the Control group. When liver and body
weight are compared collectively as a ratio, the Second Treatment
Group non-significantly trends lower than the Control or First
Treatment Group.
[0430] Blood Glucose
[0431] There was no significant difference in whole blood glucose
between the Control Group and the First Treatment Group whereas
whole blood glucose level tended to decrease in the Second
Treatment Group compared with the Control Group.
[0432] Liver Triglycerides
[0433] Liver triglyceride contents tended to decrease in the First
Treatment Group and significantly decreased in the Second Treatment
Group compared with the Control Group.
CONCLUSIONS
[0434] The study showed that animals for which treatment was
started later, the Second Treatment Group, had significantly
reduced the liver triglyceride contents compared with the control
group. Although the liver triglycerides tended to decrease in both
treatment groups, the First Treatment Group showed a
non-significant change (at 0.05) and the Second Treatment Group
showed a statistically significant decrease. This Second Treatment
Group also showed a decreasing tendency of whole blood glucose
levels compared with the Control Group. The significant decrease in
the liver triglycerides in the more progressed group demonstrate
that certain embodiments of the present invention can improve lipid
and glucose metabolism in the liver. These results also demonstrate
that omega-3 fatty acids can improve lipid and glucose metabolism
in the liver.
[0435] The effect of the treatment was not merely through an
induction of weight loss by the animals. In fact, the study found
that body weight gradually increased during the treatment period in
all groups and that there was no difference in mean body weight
between groups. Although the end liver weight of the subjects in
the treatment groups was lower than that of the subjects in the
Control Group, the difference was not statistically significant. In
addition, both treatment groups trended to reduce alpha-SMA (a
marker of myofibroblast associated with the development of liver
fibrosis) mRNA expression in the liver, a key indicator of the
treatment's demonstrated ability to delay the progression to NASH.
There were no significant differences in fibrosis area between the
Control Group and either group, data not shown.
[0436] Without being held to any one theory, we believe the study
demonstrates the ability of at least one composition disclosed
herein to deliver polyunsaturated EPA and DHA fatty acids to
mammals with NAFLD for incorporation into the cell membrane of
erythrocytes, wherein said delivery can reduce the triglyceride
level in the liver and thereby slow the disease's progression to
NASH. The study may also indicate that administration of
compositions disclosed herein might provide a larger medicinal
response when provided later in the disease progression from NAFLD
to NASH.
Neurodegenerative Diseases Prophetic Non-Limiting Working
Examples
Example 9
[0437] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 9:
TABLE-US-00009 TABLE 9 COMPOSITION INGREDIENT Amount (mg) % (wt/wt)
DHA Omega-3 fatty acid Ethyl 754.3 68.57 Ester*.dagger. Polysorbate
80 337.9 30.72 Poloxamer 237 (Pluronic .RTM. F87) 7.8 0.71 TOTAL
1100 100 *The omega-3 oil may contain ~2% .alpha.-tocopherol as an
antioxidant. .dagger.DHA comprises at least 90% of the DHA oil
(678.9 mg). The majority of the remaining no more than 10% is EPA
(~5%, or up to 37.7 mg)
Example 10
[0438] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 10:
TABLE-US-00010 TABLE 10 COMPOSITION INGREDIENT Amount (mg) %
(wt/wt) EPA/DHA Omega-3 fatty acid Ethyl 754.3 65.59 Ester*.dagger.
Gamma-tocotrienol 50.0 4.35 Polysorbate 80 337.9 29.38 Poloxamer
237 (Pluronic .RTM. F87) 7.8 0.68 TOTAL 1150 100 *The omega-3 oil
may contain ~2% .alpha.-tocopherol as an antioxidant. .dagger.EPA
comprises 383 mg and 170 mg DHA.
Primary Sclerosing Cholangitis Prophetic Non-Limiting Working
Examples
Example 11
[0439] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 11:
TABLE-US-00011 TABLE 11 COMPOSITION INGREDIENT Amount (mg) %
(wt/wt) DHA Omega-3 fatty acid Ethyl 754.3 68.57 Ester*.dagger.
Polysorbate 80 337.9 30.72 Poloxamer 237 (Pluronic .RTM. F87) 7.8
0.71 TOTAL 1100 100 *The omega-3 oil may contain ~2%
.alpha.-tocopherol as an antioxidant. .dagger.DHA comprises at
least 90% of the DHA oil (678.9 mg). The majority of the remaining
no more than 10% is EPA (~5%, or up to 37.7 mg)
Example 12
[0440] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 12:
TABLE-US-00012 TABLE 12 COMPOSITION INGREDIENT Amount (mg) %
(wt/wt) EPA/DHA Omega-3 fatty acid Ethyl 754.3 65.59 Ester*.dagger.
Gamma-tocotrienol 50.0 4.35 Polysorbate 80 337.9 29.38 Poloxamer
237 (Pluronic .RTM. F87) 7.8 0.68 TOTAL 1150 100 *The omega-3 oil
may contain ~2% .alpha.-tocopherol as an antioxidant. .dagger.EPA
comprises 383 mg and 170 mg DHA.
Example 13
[0441] The study described below will assess the effect of
compositions described herein in a murine model of Primary
Sclerosing Cholangitis (PSC). It is believed that treatment with
the compositions described herein will be more effective in
preventing and/or reversing bile duct injury in a mouse model of
PSC.
[0442] The study will utilize a mouse model of PSC induced by DSS
& DDC. The well established mouse model of PSC where exon 10
CFTR.sup.+/- knockout mice (heterozygotes) will be exposed to oral
Dextran Sodium Sulfate (DSS) followed by up to 28 days of the
xenobiotic DDC dissolved in peptamen (Martin C R, Zaman M M,
Ketwaroo G A, Bhutta A Q, Coronel E, Popov Y, Schuppan D, Freedman
S D. CFTR dysfunction predisposes to fibrotic liver disease in a
murine model. Am J Physiol Gastrointest Liver Physiol 303:
G474-G481, 2012).
[0443] The Cftr.sup.+/- mouse model of PSC consists of two phases:
(1) induction of colitis with DSS for 10 days or until overt blood
per rectum; and, (2) induction of liver inflammation and fibrosis
with DDC for 28 days. The following protocol will begin using
Cftr.sup.+/- mice at .about.age 35 days, or week 5.
[0444] Induction of Colitis with DSS for 10 Days
[0445] 85 mg of DSS dissolved in 12 ml of Peptamen by continuous
vortexing.
[0446] The mice will be fed using a liquid feeder (Dyets Inc.,
Bethlehem, Pa.).
[0447] The mice will be monitored daily for onset of colitis by
overt bleeding per rectum.
[0448] Once overt bleeding is noted (typically day 8-10 of DSS),
DSS will be discontinued and the mice will be fed peptamen alone
for one day.
[0449] Induction of Liver Inflammation and Fibrosis with Oral DDC
for 28 Days
[0450] After overt colitis and one day of peptamen feeding alone,
the mice will be treated for 28 days with 1.5 mg DDC in 12 ml of
Peptamen per day
[0451] Volume consumed will be recorded.
[0452] Treatment Groups
[0453] 3 treatment groups will be used:
1) High DHA (40 mg/day) 2) High DHA (40 mg/day) with high
bioavailability 3) No treatment
[0454] All treatments will begin 3 days prior to the administration
of DSS and thus will examine prevention of injury.
[0455] Sample Collection and Outcomes Measures
a) Mice will be sacrificed by exposure to CO.sub.2 for 2 min. b)
Body weight will be measured at the start of the protocol, every
3.sup.rd day, then at the end of the protocol. Percent weight
change will be calculated as follows: i.=((Starting weight-End
weight)/Starting weight)*100 c) Liver and blood samples will be
collected at sacrifice and stored as described below in Table 13
below. d) Fatty acid levels will be determined in plasma and
liver.
TABLE-US-00013 TABLE 13 Sample collection and processing Outcome
Measure Sample Sample preparation/storage Sample analysis Tissue
Weight Liver Liver removed and weighed pre- BIDMC fixation
Histology Liver Liver fixed in 10% buffered BIDMC for blind
formalin for 2 hours followed by scoring two PBS washes and stored
in PBS at 4.degree. C. H&E staining, Sirius red staining Fatty
Acids Liver Fatty acid extraction BIDMC GC-MS Serum Blood will be
collected by cardiac puncture using a 1 ml syringe equipped with a
25 G needle. Blood will be placed on ice for 15 minutes. The sample
will be centrifuged at 5500Xg and serum collected. On average,
150-200 uL of serum will be collected per mouse. The serum will be
pre-aliquoted into separate vials for the following distribution.
All aliquoted samples will be stored at -80.degree. C. Fatty Acids
Serum Fatty acid extraction BIDMC GC-MS The following will be
collected and stored for possible subsequent analyses depending on
histology results. LOXL-2, .alpha.-SMA, or Liver
Immunohistochemistry BIDMC other markers TBD Two separate pieces of
liver tissue will be provided for each animal. Tissue will be
snap/flash frozen in tissue biopsy cassettes using liquid nitrogen
and will be store at -80.degree. C. Inflammatory/Fibrosis Liver
Snap freeze, store at -80.degree. C. RT PCR at BIDMC markers:
procollagen-1, LOXL-2, TGF-.beta.1, TGF-.beta.2, TIMP-1, TIMP-2,
MMP-2, MMP-9, TNF-.alpha., .alpha.-SMA Liver Function Tests: Sample
Amount Destination Alkaline Phosphatase, Serum 50 uL BIDMC ALT,
AST, Bilirubin Luminex Cytokine Serum 70 uL BIDMC Panel
Tissue Histological Slide Scoring
[0456] a) Slides will prepared from the left lateral and bifurcated
medial lobes of the liver; each liver will have one slide stained
with H&E and one with Sirius Red respectively. b) Liver injury
will be scored based on four criteria: (1) epithelial injury,
intraepithelial inflammation, and mononuclear cell infiltration;
(2) bile duct proliferation; (3) bile duct angulation; and (4)
fibrosis. Each criterion receives a score between 0 and 3. A
scoring rubric is defined in the box below. Scoring as previously
described in our AJP article, will be performed by two individuals
blinded to the conditions
TABLE-US-00014 SCORING RUBRIC 0 Absent 1 Mild 2 Moderate 3
Severe
i. Epithelial injury, intraepithelial inflammation, and mononuclear
cell infiltration will be scored on slides at 20.times.
magnification stained with H&E. ii. Bile duct proliferation
will be scored at 5.times. magnification on slides stained with
H&E. iii. Bile duct angulation will be scored at 20.times.
magnification on slides stained with H&E. iv. Fibrosis will be
scored on slides at 5.times. magnification stained with Sirius Red.
All tissues will be held and processed at the same time for Sirius
Red staining.
[0457] Fibrosis will be quantified on Sirius Red slides using
ImageJ
Sickle Cell Disease Prophetic Non-Limiting Working Examples
Example 14
[0458] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 14:
TABLE-US-00015 TABLE 14 COMPOSITION INGREDIENT Amount (mg) %
(wt/wt) DHA Omega-3 fatty acid Ethyl 754.3 68.57 Ester*.dagger.
Polysorbate 80 337.9 30.72 Poloxamer 237 (Pluronic .RTM. F87) 7.8
0.71 TOTAL 1100 100 *The omega-3 oil may contain ~2%
.alpha.-tocopherol as an antioxidant. .dagger.DHA comprises at
least 90% of the DHA oil (678.9 mg). The majority of the remaining
no more than 10% is EPA (~5%, or up to 37.7 mg)
Example 15
[0459] The amounts and percentages of the ingredients comprising
one embodiment of the compositions described herein are shown in
Table 15:
TABLE-US-00016 TABLE 15 COMPOSITION INGREDIENT Amount (mg) %
(wt/wt) EPA/DHA Omega-3 fatty acid Ethyl 754.3 65.59 Ester*.dagger.
Gamma-tocotrienol 50.0 4.35 Polysorbate 80 337.9 29.38 Poloxamer
237 (Pluronic .RTM. F87) 7.8 0.68 TOTAL 1150 100 *The omega-3 oil
may contain ~2% .alpha.-tocopherol as an antioxidant, .dagger.EPA
comprises 383 mg and 170 mg DHA.
Example 16
[0460] A Randomized, double blind, placebo-controlled study will be
conducted to investigate the therapeutic potential of the
compositions described herein for patients with homozygous sickle
cell disease.
[0461] Objective: [0462] 1. To evaluate efficacy of the
compositions described herein in treatment of sickle cell anemia.
[0463] 2. To study change in number of sickle cell vaso-occlusive
crises. [0464] 3. To monitor the safety of one or more of the
compositions described herein in the treatment of sickle cell
anemia [0465] Sample size: 60
[0466] Study medication (n=sample in each group): [0467] 1. Test
(T): One or more of the compositions described herein (n=30) [0468]
2. Reference (R): Standard of Care (n=30)
[0469] Duration of treatment: 6 months
[0470] Inclusion Criteria: [0471] 1. Patients aged 2-24 y with
HbSS, who are undergoing regular follow-up as an outpatient with
Sickle Cell Disease. [0472] 2. The patients who will be in a steady
state, defined as no evidence of fever, infection, or crisis for 4
wk before the start of the study. [0473] 3. Phenotypic
characteristic was confirmed with the use of cellulose acetate
electrophoresis at pH 8.5. [0474] 4. Can continue standard of care
treatment. [0475] 5. Already on fish oil compounds will require
wash out of 15 days.
[0476] Exclusion Criteria: [0477] 1. Presence of other chronic
diseases. [0478] 2. Blood transfusion in the previous 4 mo. [0479]
3. A history of overt stroke. [0480] 4. Pregnancy.
[0481] Medication permitted: All of the patients receiving regular
folate supplementation, and those, 5 y receiving standard oral
prophylactic penicillin.
[0482] Investigation Parameters: Blood count, inflammatory markers
(C-RP)
[0483] The primary end point will be rates of clinical
vaso-occlusive crisis and secondary end points will include number
of hospitalizations, haemolytic events, blood transfusion rate,
school attendance, and blood count--will be analysed by
intention-to-treat analysis and quality of life assessment.
[0484] Statistics: N=30. The vaso-occlusive crises will be
summarized on the basis of the annualized crisis rate by dividing
the total number of crises experienced by the number of follow-up
months and multiplying by 12.
Example 17
[0485] The amounts and percentages of the ingredients comprising
one embodiment of the composition, also referred to as SC401, are
shown in Table 17:
TABLE-US-00017 TABLE 17 INGREDIENT Amount (mg) % (wt/wt)
COMPOSITION (FILL MASS)/dosage form Total Omega-3 fatty acid Ethyl
Esters 754.3 68.57 EPA Ethyl Esters 392.2 35.65 DHA Ethyl Esters
165.9 15.08 Polysorbate 80 337.9 30.72 Poloxamer 237 (Pluronic
.RTM. F87) 7.8 0.71 GEL MASS/dosage form Gelatin 270 40 Glycerin
135 20 Purified water 270 40
[0486] The pharmacokinetic (PK) profiles and bioavailability of EPA
and DHA comprising the SC401 formulation is graphically represented
in FIGS. 10A, 10B, 11A AND 11B.
[0487] FIG. 10 shows mean EPA and DHA total lipid plasma
concentration profiles (.mu.g/ml) (baseline-adjusted) after
administration of a single dose (dose adjusted) of SC401 and
Lovaza.RTM. in fasted conditions.
* * * * *