U.S. patent application number 10/950734 was filed with the patent office on 2005-04-28 for arginyl-glutamine dipeptide for treatment of pathological vascular proliferation.
Invention is credited to Anthony, Joshua C., Grant, Maria B., Neu, Josef.
Application Number | 20050089547 10/950734 |
Document ID | / |
Family ID | 34393151 |
Filed Date | 2005-04-28 |
United States Patent
Application |
20050089547 |
Kind Code |
A1 |
Neu, Josef ; et al. |
April 28, 2005 |
Arginyl-glutamine dipeptide for treatment of pathological vascular
proliferation
Abstract
The subject invention provides dipeptides useful in preventing
pathological proliferation of blood vessels. The dipeptides of the
subject invention are particularly advantageous because they are
stable, bioavailable, and can be formulated in an aqueous solution,
and in particular, into infant formulations.
Inventors: |
Neu, Josef; (Gainesville,
FL) ; Grant, Maria B.; (Fairfield, FL) ;
Anthony, Joshua C.; (Evansville, IN) |
Correspondence
Address: |
Nelson Mullins Riley & Scarborough, LLP
Meridian
Suite 1700
1320 Main Street
Columbia
SC
29201
US
|
Family ID: |
34393151 |
Appl. No.: |
10/950734 |
Filed: |
September 27, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60506413 |
Sep 26, 2003 |
|
|
|
Current U.S.
Class: |
424/439 ;
514/21.91; 514/5.5 |
Current CPC
Class: |
A61P 9/10 20180101; A61K
38/16 20130101; A61K 38/04 20130101; A61K 38/05 20130101; A23L
33/18 20160801 |
Class at
Publication: |
424/439 ;
514/019 |
International
Class: |
A61K 038/04; A61K
047/00 |
Claims
What is claimed is:
1. An infant formula comprising a source of arginyl-glutamine
dipeptide.
2. The infant formula according to claim 1, wherein the source of
arginyl-glutamine dipeptide is arginyl-glutamine dipeptide, or a
precursor thereof.
3. The infant formula according to claim 1, wherein the infant
formula comprises arginyl-glutamine dipeptide, or a precursor
thereof, in an amount sufficient for the prevention or treatment of
a condition that is selected from retinopathy of prematurity,
diabetic retinopathy, vascular proliferative retinopathy, or
proliferation of abnormal vascularization.
4. The infant formula according to claim 1, wherein the infant
formula is nutritionally complete.
5. The infant formula according to claim 3, wherein the infant
formula contains arginyl-glutamine dipeptide, or precursor thereof,
in an amount less than 0.1% by weight.
6. The infant formula according to claim 3, which contains an
amount of arginyl-glutamine dipeptide of from about 0.001% to
0.098% by weight of the formula.
7. The infant formula according to claim 2, wherein the source of
arginyl-glutamine dipeptide is one or more of arginyl-glutamine
containing oligopeptides, polypeptides, peptides, or proteins.
8. A method for the prevention or treatment of a condition that is
selected from retinopathy of prematurity, diabetic retinopathy,
vascular proliferative retinopathy, or proliferation of abnormal
vascularization, in an infant, the method comprising administering
to the infant an infant formula or dietary supplement containing
arginyl-glutamine dipeptide, or precursor thereof, in an amount
that is sufficient to treat or prevent the condition.
9. The method according to claim 8, wherein administering comprises
administering a nutritionally complete infant formula.
10. The method according to claim 8, wherein the infant is
administered an amount of the formula or supplement sufficient to
provide the infant the arginyl-glutamine dipeptide in an amount of
from about 0.001 to about 10,000 mg/kg of body weight of the infant
per day.
11. The method according to claim 8, wherein infant is administered
an amount of the formula or supplement sufficient to provide the
infant the arginyl-glutamine dipeptide in an amount of from about
0.001 to about 100 mg/kg of body weight of the infant per day.
12. The method according to claim 8, wherein infant is administered
an amount of the formula or supplement sufficient to provide the
infant the arginyl-glutamine dipeptide in an amount of from about
0.01 to about 1 mg/kg of body weight of the infant per day.
13. The method according to claim 8, wherein the infant is a
premature infant.
14. The method according to claim 8, wherein the infant is a
newborn infant.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. provisional
application Ser. No. 60/506,413, filed Sep. 26, 2003.
BACKGROUND OF THE INVENTION
[0002] People suffering from visual impairment face many challenges
in performing routine daily activities and/or may not be able to
fully enjoy the visual aspects of their surroundings. Of particular
concern with regard to the current invention are visual impairments
caused by damage to the retina, which occur in conditions such as
diabetic retinopathy and retinopathy of prematurity.
[0003] Diabetic retinopathy is a progressive disease characterized
by abnormalities of the blood vessels of the retina caused by
diabetes, such as weakening of the blood vessel walls, leakage from
the blood vessels, and bleeding and scarring around new vessels.
Diabetic retinopathy results in impairment of a person's vision
causing severely blurred vision and, potentially, blindness.
[0004] Diabetes affects over 16 million Americans. The World Health
Organization indicates that diabetes afflicts 120 million people
worldwide, and estimates that this number will increase to 300
million by the year 2025. Diabetics are faced with numerous
complications including kidney failure, non-traumatic amputations,
an increase in the incidence of heart attack or stroke, nerve
damage, and loss of vision. Diabetic retinopathy is a form of
visual impairment often suffered by diabetics.
[0005] Due to significant medical advancements, diabetics are able
to live much longer than in the past. However, the longer a person
has diabetes the greater the chances of developing diabetic
retinopathy. Affecting over 5.3 million Americans, diabetic
retinopathy is the leading cause of blindness among adults in the
United States. Annually, in the United States, between 12,000 and
24,000 people lose their sight because of diabetes.
[0006] While management of diabetic retinopathy has improved, risk
of complications, such as loss of visual acuity, loss of night
vision and loss of peripheral vision, remains significant and
treatment sometimes fails. Currently, laser photocoagulation is the
most effective form of therapy for advanced disease. Unfortunately,
current treatment options are inadequate and the disease is often
progressive even with successful glucose control.
[0007] Retinopathy of prematurity (ROP) is a disorder of retinal
blood vessel development in the premature infant. Under normal
development, blood vessels grow from the back central part of the
eye out toward the edges. In premature babies, this process is not
complete and the abnormal growth of the vessels proliferate leading
to scar tissue development, retinal detachment and possibly
complete blindness.
[0008] ROP is the major cause of blindness in children under the
age of 7. The salient pathological features are neovascularization
in the retinal vascular endothelium with edema and breakdown in the
blood-retinal barrier (BRB) that leads to hemorrhage, tissue damage
and retinal scarring ultimately leads, in the severest cases, to
blindness.
[0009] Improved care in the neonatal intensive care unit has
reduced the incidence of retinopathy of prematurity in moderately
premature infants. Ironically, however, increasing rates of
survival of very premature infants, who would have had little
chance of survival in the past, has increased the occurrence of
retinopathy of prematurity. Since these very premature infants are
at the highest risk of developing ROP, it is of great concern that
the condition may actually be becoming more prevalent again.
[0010] For those babies in whom retinopathy progresses, treatment
is necessary. Cryotherapy and laser treatment have some effect in
advanced stages of the disease, saving a degree of vision in a
proportion of the eyes that would otherwise have been blinded, but
prevention awaits a better understanding of major causative factors
and underlying pathophysiology.
[0011] Current research shows promise that the prevention of
retinal blood vessel damage, which marks retinopathy, may be
achieved by the utilization of certain compounds. It has been
demonstrated that, in retinal epithelial cells, glutamine
deprivation can lead to upregulation of vascular endothelial growth
factor (VEGF) expression (Abcouwer S. et al., "Response of VEGF
expression to amino acid deprivation and inducers of endoplasmic
reticulum stress", Invest Ophthalmol Vis Sci, August 2002, pp.
2791-8, Vol. 43, No. 8). Most sick premature infants are deprived
of glutamine during the time they receive supplemental oxygen, a
known predisposing factor in the development of ROP. The over
expression of VEGF during this time period is also thought to be
involed in the pathogenesis of ROP providing glutamine supplements
during this time period could potentially down-regulate VEGF.
Arginine is substrate for the reaction that produces nitric oxide,
a very potent vasodilator, vasodilation in retinal blood vessels
also prevents neovascularization. Nitirc oxide also has numerous
other beneficial effects and is now commonly used for treatment of
lung disease in critically ill infants.
[0012] It is well known that proteins are converted to amino acids
in the digestive system and that the resulting amino acids are used
by the body for growth and development. Proteins and peptides
administered for therapeutic or preventative measures are also
well-known. Oligopeptides are better absorbed in the intestines
than individual amino acids.
[0013] European Patent Application No. 0,182,356 discloses a
nutritional composition containing at least one oligopeptide
consisting of a dipeptide or a tripeptide wherein the N-terminal
amino acid residue is selected from the class consisting of
alanine, lysine and arginine.
[0014] One group conducting research in this area concluded that
glycine is generally superior to other amino acids as the
N-terminal amino acid residue in a dipeptide. This superiority was
attributed to a greater fraction of such an intravenously
administered dipeptide reaches the tissues. S. Adibi et al.,
Influence of Molecular Structure on Half-life and Hydrolysis of
Dipeptides in Plasma: Importance of Glycine as N-Terminal Amino
Acid Residue, 35 Metabolism 850, 835 (1986).
[0015] Two European patents, 0,087,751 and 0,087,750 disclose
water-soluble peptides. The '751 patent discloses a method to
parenterally administer low water-soluble amino acids. Two amino
acids, tyrosine and cystine, individually have low solubility in
water. These amino acids, however, are clinically useful and,
therefore, it was desirable to find an effective formulation. The
'751 patent describes an infusion method which involves bonding
these relatively insoluble amino acids to the amino acid lysine to
produce a tripeptide.
[0016] The '750 patent discloses the infusion of glutamine as a
derivative substituted by .alpha.-aminoacyl residues on the a amino
group. That is, glutamine is in the "c-terminal" position, in that
its alpha amino nitrogen becomes part of the peptide bond with the
other amino acid. The preferred dipeptide preparation disclosed in
the '750 patent is alanyl-glutamine. The aminoacylation of
glutamine is reported to achieve a stabilization of the terminal
amide group.
[0017] Experiments involving the use of total parenteral nutrition
(TPN) containing glycyl-glutamine dipeptides, however, suggest
potential adverse effects of the TPN formulation containing
glycyl-glutamine (U.S. Pat. No. 5,189,016).
[0018] Also, the use of an arginyl-glutamine dipeptide for the
prevention of muscle breakdown and microbial infections has
recently been described. See, WO 03/017787. These amino acids have
also been described in complex compositions (Miyazawa et al. (1976)
Journal of Faculty of Fisheries and Animal Husbandry Hiroshima
15(2):161-169; and JP 2119762).
[0019] Two commercially available dipeptides of glutamine are
Dipeptiven, which is an alanyl-glutamine (Fresenius Laboratories,
Germany) and Glamin (Pharmacia and Upjohn Laboratory, Sweden),
which is an amino acid solution containing glycyl-glutamine
dipeptide. To this date, there are no published studies of the
arginyl-L-glutamine dipeptide.
[0020] With the increase of adult onset diabetes, longer life span
for diabetics and high rate of survival of very premature infants,
many individuals are now at even greater risk for developing
retinopathy. Although treatment options, such as laser therapy,
exist for both conditions, the results are inadequate and the
disease often remains progressive. There remains a great need in
the art for compositions and methods which prevent and/or treat
retinal diseases.
BRIEF SUMMARY OF THE INVENTION
[0021] The subject invention provides materials and methods useful
in preventing proliferation of abnormal blood vessels. The
prevention of the over-proliferation of these blood vessels
according to the subject invention is particularly advantageous for
treatment of certain ocular conditions including premature infants
at risk for retinopathy of prematurity and individuals at risk for
diabetic retinopathy.
[0022] In a preferred embodiment of the subject invention, the
amino acids arginine and glutamine are combined as the dipeptide
arginyl-glutamine in order to provide beneficial effects in a safe,
easily absorbable formulation. The dipeptide of the subject
invention is particularly advantageous because the solubility of
the dipeptide is greater than the individual amino acids.
[0023] Advantageously, the dipeptide of the subject invention
inhibits the over-proliferation of unwanted blood vessels. The
dipeptide of the subject invention is also advantageous because it
is safe for human and animal use and can be readily formulated in
an aqueous solution.
[0024] The compounds of the subject invention can be formulated
according to known methods for preparing pharmaceutically useful
compositions. Formulations are described in detail in a number of
sources which are well known and readily available to those skilled
in the art. In general, the compositions of the subject invention
will be formulated such that an effective amount of the bioactive
compound(s) is combined with a suitable carrier in order to
facilitate effective administration of the composition.
[0025] In accordance with the invention, pharmaceutical
compositions comprising, as an active ingredient, an effective
amount of one or more of the compounds and one or more non-toxic,
pharmaceutically acceptable carriers or diluents. Pharmaceutical
carriers or excipients may contain inert ingredients which do not
interact with the compound, or ingredients that do interact with
the compound but not in a fashion so as to interfere with the
desired effect. In general, the formulations are prepared by
uniformly and intimately bringing into association the active
ingredient with liquid carriers or finely divided solid carriers or
both, and then, if necessary, shaping the product. Examples of such
carriers for use in the invention include ethanol, dimethyl
sulfoxide, glycerol, silica, alumina, starch, and equivalent
carriers and diluents.
[0026] The dipeptide of the subject invention can also be
formulated as a nutraceutical, including drinks, drink mixes, and
bars.
BRIEF SUMMARY OF THE FIGURES
[0027] FIG. 1 shows inhibition of neovascularization by an
arginyl-glutamine dipeptide in a model of retinopathy of
prematurity; and
[0028] FIG. 2 shows inhibition of neovascularization by an
arginyl-glutamine dipeptide in a model of retinopathy of
prematurity at various concentrations of the dipeptide.
DETAILED DISCLOSURE OF THE INVENTION
[0029] The present invention provides compositions containing
therapeutic dipeptides and methods for administering the same. The
subject invention provides a novel, safe and affordable therapy for
treatment of pathological ocular vascular proliferation.
Advantageously, the subject invention provides a dipeptide having
excellent water solubility, stability to sterilization, long-term
stability, and bioavailability for humans and animals. One
embodiment of the present invention provides a composition
comprising an aqueous pharmaceutical solution having at least one
arginyl-glutamine dipeptide. Other embodiments include
nutraceutical formulations.
[0030] In a specific embodiment of the subject invention the
arginyl-glutamine dipeptides described herein can be used for
preventing the proliferation of abnormal retinal blood vessels in a
patient. Thus, these dipeptides can be administered to premature
infants or diabetics who are at risk for retinal disease.
[0031] In accordance with the teachings provided herein, aqueous
clinical compositions can be prepared that include at least one
arginyl-glutamine dipeptide. The dipeptide can be added to enteral
or parenteral formulations. Each dipeptide has an N-terminal amino
acid which is arginine. The C-terminal amino acid is glutamine.
[0032] The concentration of the dipeptide in the aqueous solution
can be, for example, from about 0.1 to about 25.0 percent by
weight. As discussed in more detail below, in addition to the
dipeptides of the subject invention, the clinical solution can
contain, for example, dextrose, liquid emulsions, vitamins,
minerals and trace elements. The selection of the particular
dipeptide formulation depends upon the particular use.
[0033] For parenteral administration, a supply of the dipeptide
solution may be merged through a Y-connection with a supply of
glucose solution or other parenteral solutions. The dipeptide
solutions may also be mixed with glucose solutions and/or other
parenteral solutions to create a mixture which may be administered
parenterally.
[0034] The administration of dipeptides rather than free amino
acids allows administration of the same amount of amino acid
residue in solutions which are less hypertonic and therefore can be
introduced into peripheral veins.
[0035] The dipeptides of the subject invention can be readily
synthesized and/or formulated by a person skilled in the art having
the benefit of the instant disclosure. Alternatively, the
dipeptides can be purchased commercially from, for example, Bachem
Biosciences, Inc. which sells the H-Arg-Glu-OH salt.
[0036] The subject invention contemplates the administration of the
Arg-Glu dipeptide in any appropriate formulation including, for
example, salts, prodrugs, and extended release formulations (such
as, for example, formulation with polyethylene glycol (PEG)). The
peptide itself may be administered, as well as oligopeptides,
peptides, proteins, protein hydrolyzates, and any other materials
that could serve as a source of the dipeptide.
[0037] In the case of oligopeptides, peptides, and proteins, these
prodrug formulations may be designed with, for example, cleavage
sites adjacent to each side of the dipeptide so that the dipeptide
is generated upon exposure to enzymes, acids or other factors. In
one embodiment, a polypeptide can be prepared with multiple
dipeptides separated by cleavage sites, thereby creating multiple
dipeptides upon exposure to the cleaving factor. This cleaving to
create the dipeptide can be done as part of a production process or
in vivo as the result of, for example, digestive enzymes and/or
acids. The source of the dipeptide may be natural or the dipeptide
(including prodrugs) may be produced recombinantly or
synthetically.
[0038] In one method, the subject invention involves identifying an
individual who has, or who is at risk for developing pathological
vascularization and then providing that individual with a
composition comprising the dipeptide of the subject invention (or a
prodrug thereof) along with instructions or information concerning
the activity of the dipeptide to inhibit pathological
vascularization. In one embodiment, a nutriceutical is provided
that has the dipeptide, and/or a prodrug thereof, along with
instructions for consuming the nutriceutical to promote
cardiovascular (or just vascular) health.
[0039] The compounds of the invention are useful for various
therapeutic purposes. Specifically, as described herein, the
compounds of the invention are effective for inhibiting vascular
retinopathy and other forms of pathological vascular proliferation.
Accordingly, these compounds are useful prophylactically and
therapeutically for treating animals and humans at risk for
pathological vascular proliferation including vascular retinopathy
and vasculature associated with tumors.
[0040] Therapeutic application of the compounds and compositions
containing them can be accomplished by any suitable therapeutic
method and technique presently or prospectively known to those
skilled in the art.
[0041] The peptides provided by the present invention are typically
administered to a mammal, particularly a human, dog or cat, any of
which is intended to be encompassed by the term "patient" herein,
in need of the prevention or treatment of pathological vascular
proliferation. Pathological conditions involving vascular
proliferation include, for example, tumor growth, age-related
macular degeneration, vascular proliferation associated with
angioplasty and/or stents, diabetic retinopathy and retinopathy of
prematurity. Thus, the dipeptides of the subject invention can be
used to treat angiogenic diseases. Angiogenic diseases include
those that are disclosed in U.S. Pat. No. 5,759,547, which is
incorporated herein, in its entirety, by reference.
[0042] Angiogenesis and neovascularization in the adult animal is
usually a pathological process, and is in direct contradistinction
to non-pathological neovascularization, which usually occurs in
normal embryogenesis (e.g., development of the embryonic vascular
system). In accordance with the subject invention,
neovascularization refers specifically to pathological
neovascularization. Aberrant or pathological vascularization is a
key component in numerous disease states. For example,
vascularization is a critical element of most solid tumors, such as
cancers of the brain, genitourinary tract, lymphatic system,
stomach, larynx and lung. These include histiocytic lymphoma, lung
adenocarcinoma and small cell lung cancers. Aberrant vascular
growth in the retina can lead to visual degeneration which can
culminate in blindness. Accordingly, the subject invention provides
dipeptide compounds and formulations thereof for the treatment of
neovascularization.
[0043] Compounds of the invention can also be used to inhibit the
proliferation of vascular endothelial cells and so are indicated
for use in treating graft vessel diseases such as restenosis or
vascular occlusion following vascular insult such as angioplasty,
allo- or xenotransplant vasculopathies, graft vessel
atherosclerosis, and in the transplantation of an organ (e.g.,
heart, liver, lung, kidney or pancreatic transplants (Weckbecker et
al., Transplantation Proceedings 1997, 29, 2599-2600).
[0044] The peptides are administered by incorporating the peptide
in a pharmaceutical composition comprising the peptide or a
non-toxic pharmaceutically acceptable salt or prodrug thereof and a
non-toxic pharmaceutically acceptable carrier therefor.
[0045] The peptide or its salt or prodrug is employed in an
effective amount i.e. an amount sufficient to evoke the desired
pharmacological response. This is generally an amount sufficient to
produce lessening of one or more of the effects of pathological
vascular proliferation. In the case of retinopathy, it is an amount
sufficient to produce regression of neovascularization and/or an
amount sufficient to produce improved visual acuity.
[0046] The terms "pharmaceutically acceptable carrier" or a
"carrier" refer to any generally acceptable excipient or drug
delivery device that is relatively inert and non-toxic. The peptide
can be administered with or without a carrier. When treating
retinopathies, a preferred embodiment is to administer the peptide
to the retinal area or the vasculature around or leading to the
retina. Exemplary carriers include calcium carbonate, sucrose,
dextrose, mannose, albumin, starch, cellulose, silica gel,
polyethylene glycol (PEG), dried skim milk, rice flour, magnesium
stearate, and the like. The peptide can be administered
systemically or locally (e.g., by injection or diffusion). Suitable
carriers (e.g., pharmaceutical carriers) also include, but are not
limited to sterile water, salt solutions (such as Ringer's
solution), alcohols, polyethylene glycols, gelatin, carbohydrates
such as lactose, amylose or starch, magnesium stearate, talc,
silicic acid, viscous paraffin, fatty acid esters,
hydroxymethylcellulose, polyvinyl pyrolidone, etc. Such
preparations can be sterilized and, if desired, mixed with
auxiliary agents, e.g., lubricants, preservatives, stabilizers,
wetting agents, emulsifiers, salts for influencing osmotic
pressure, buffers, coloring, and/or aromatic substances and the
like which do not deleteriously react with the active compounds.
They can also be combined where desired with other active
substances, e.g., enzyme inhibitors, to reduce metabolic
degradation. A carrier (e.g., a pharmaceutically acceptable
carrier) is preferred, but not necessary to administer the
peptide.
[0047] Suitable non-toxic pharmaceutically acceptable carriers for
use with the peptide will be apparent to those skilled in the art
of pharmaceutical formulation. See, for example, Remington's
Pharmaceutical Sciences, seventeenth edition, ed. Alfonso R.
Gennaro, Mack Publishing Company, Easton, Pa. (1985). The choice of
suitable carriers will depend upon the exact nature of the
particular dosage form selected.
[0048] The supplement can take on various forms, including but not
limited to pills, edible bars, drinks or drink mix. The compounds
of the subject invention may be combined with other components such
as, for example, a soluble fiber compound. The soluble fiber
compound may be, for example, locust gum, guar gum, pectin, gum
arabic, or psyllium.
[0049] The person skilled in this art, having the benefit of the
current disclosure can readily formulate the compounds of the
subject invention into a pill, bar, or other edible composition for
easy and enjoyable consumption. These therapeutic compositions can
be used as described herein.
[0050] In one embodiment, the present invention provides a
nutriceutical composition. The dipeptides can be obtained or
produced using processes known to those skilled in the art having
the benefit of the instant disclosure.
[0051] The compositions of the invention are useful for various
therapeutic purposes. Because of the antiproliferative properties
of the compounds, they are useful to prevent unwanted cell growth
in a wide variety of settings including in vitro uses.
[0052] Therapeutic application of the new compositions can be
accomplished by any suitable therapeutic method and technique
presently or prospectively known to those skilled in the art.
[0053] When used to reduce the severity or incidence of tumors, the
dosage administration to a host in the above indications will be
dependent upon the identity of the cancer cells, the type of host
involved, its age, weight, health, kind of concurrent treatment, if
any, frequency of treatment, and therapeutic ratio.
[0054] In a preferred embodiment, the active ingredient at the site
of treatment is the dipeptide of the subject invention in the
absence of other amino acids. Thus, it is not necessary for the
subject invention to be specifically formulated with, for example,
an essential amino acid and a non-essential amino acid.
[0055] In one embodiment, the dipeptide of the subject invention
can be administered as a nutriceutical supplement in unit dosage
form. In such form, the preparation is subdivided into unit doses
containing appropriate quantities of the nutriceutically active
component. The unit dosage form can be a packaged preparation, such
as packeted tablets, capsules, lotions, ointments and powders in
vials or ampoules. Also, the unit dosage can be a drink (such as a
powder-based drink, shake or tea formulation), yogurt, solid food
product, capsule, tablet, lotion, ointment, chewing gum, lozenge,
or it can be the appropriate number of any of these in packaged
form.
[0056] In one embodiment, there is provided a nutriceutical of the
present invention in the form of a food bar. The food bar, which
may be cooked or non-cooked, may contain a grain or grains, nuts,
possibly dried fruit, sweeteners and other ingredients that may be
mixed with a binder, such as a sugar syrup or shortening, and
formed into "bars" of desirable size. Generally, food bars are
prepared in bar form of a manageable size for a product of this
nature, and bearing in mind both package size and time required for
consumption, weigh in the range of 2-3 ounces.
[0057] The food bar of the present invention may contain an amount
of protein in addition to significant amounts of complex and simple
carbohydrate, such as those non-cooked food bars described by U.S.
Pat. No. 4,055,669. Further, the food bar of the present invention
may contain dietary fiber to aid in the normalization of bowel
function and reduce the risk of colonic diseases. There are
presently available a number of non-cooked food bars that provide
varying amounts of dietary fiber while possessing requisite
qualities of acceptable taste and texture, including food bars
described in U.S. Pat. Nos. 4,673,578 and 4,871,557.
[0058] The therapeutic dosage range can be determined by one
skilled in the art having the benefit of the current disclosure.
Naturally, such therapeutic dosage ranges will vary with the size,
species and physical condition of the patient, the severity of the
patient's medical condition, the particular dosage form employed,
the route of administration and the like. In addition, a route of
administration may be selected to slowly release the chemical,
e.g., slow intravenous infusion. The inventive composition that
contains the subject dipeptide may be a nutritional composition
(nutritionally complete or nutritional supplement) for enteral
administration. That is, it is designed for oral, intragastric, or
transpyloric use.
[0059] The composition of the invention may be an infant formula or
adult nutritional composition that can be milk-based, soy-based, or
based on other food sources. The composition may be prepared as a
powder or liquid nutritional composition for formulas prepared for
infant, pediatric and adult populations. The inventive composition
may be prepared as a nutritionally complete diet by including
vitamins and minerals at acceptable levels. The subject composition
can be in the form of a dietary product such as an infant formula,
milk substitute, and meal replacement or supplement.
[0060] An embodiment of the invention is a dietary supplement that
contains arginyl-glutamine dipeptide, or a precursor thereof (which
may also be referred to herein as a prodrug). The dietary
supplement is designed to be administered along with a food or
nutritional composition, such as infant formula, and can either be
intermixed with the food or nutritional composition prior to
ingestion by the subject, or can be administered to the subject
either before or after ingestion of a food or nutritional
composition. The subject dietary supplement contains an amount of
arginyl-glutamine dipeptide, or a precursor thereof, that is
effective for the prevention or treatment of retionoathy of
prematurity, diabetic retinopathy, vascular proliferative
retinopathy, or proliferation of abnormal vascularization, and the
like.
[0061] In one embodiment, a novel infant formula containing
arginyl-glutamine dipeptide, or precursor thereof, is nutritionally
complete. By the term "nutritionally complete" is meant that the
composition contains adequate nutrients to sustain healthy human
life for extended periods. The infant formula of the invention
contains ingredients which are designed to meet the nutritional
needs of the human infant namely, a protein, carbohydrate and lipid
source and other nutrients such as vitamins and minerals.
[0062] Besides the subject dipeptide, the composition of the
invention contains a nitrogen source (i.e., amino acids and/or
protein) in an amount that is typically about 1 g to about 10 g per
100 kcal of total composition, preferably about 2 g to about 6 g
per 100 kcal; the amount of lipid source per 100 kcal of total
composition is typically greater than 0 g up to about 6 g,
preferably about 0.5 g to about 5.5 g and more preferably about 2 g
to about 5.5 g; and the amount of non-fiber carbohydrate source per
100 kcal of total composition is typically about 5 g to about 20 g,
preferably about 7.5 g to about 15 g. The amount of vitamins and
minerals in the nutritionally complete composition is typically
sufficient to meet 100% of the U.S. recommended daily intake (RDI)
in about 500 to about 3,000 kcal, preferable is about 1,000 to
about 3,000 kcal.
[0063] In one embodiment of the present nutritional composition the
amount of vitamins and minerals is sufficient to meet 100% of the
RDI in about 500 to about 3,000 kcal, preferably in about 1,000 to
about 3,000 kcal. As used herein, the RDI's are intended to mean
those published in the Federal Register, Vol. 58, No. 3, Wednesday,
Jan. 6, 1993, page 2227 which are as follows: Vitamin A, 5,000
International Units; Vitamin C, 60 milligrams; Thiamin, 1.5
milligrams; Riboflavin, 1.7 milligrams; Niacin, 20 milligrams;
Calcium, 1.0 gram; Iron, 18 milligrams; Vitamin D, 400
International Units; Vitamin E, 30 International Units; Vitamin
B.sub.6, 2.0 milligrams; Folic acid, 0.4 milligrams; Vitamin
B.sub.12, 6 micrograms; Phosphorus, 1.0 gram; Iodine, 150
micrograms; Magnesium, 400 milligrams; Zinc, 15 milligrams; Copper,
2 milligrams; Biotin, 0.3 milligram; Pantothenic acid, 10
milligrams.
[0064] In one embodiment, the novel infant formula contains the
arginyl-glutamine dipeptide, or a precursor thereof, in an amount
that is less than 0.1% by weight of the formula. It is preferred
that the amount of arginyl-glutamine dipeptide in the formula is
from about 0.001% to 0.098% by weight of the formula, more
preferred is an amount of from about 0.01% to 0.098% by weight.
[0065] In the present method, the subject infant formula or dietary
supplement is administered to an infant in an amount that is
sufficient to prevent or treat retionoathy of prematurity, diabetic
retinopathy, vascular proliferative retinopathy, or proliferation
of abnormal vascularization. In a preferred embodiment, that amount
is from about 0.001 to about 10,000 mg/kg of body weight of the
subject per day, more preferred is an amount of from about 0.001 to
about 100 mg/kg/day, yet more preferred is about 0.01 to about 50
mg/kg/day, and even more preferred is about 0.01 to about 10
mg/kg/day, and yet more preferred is about 0.01 to about 1
mg/kg/day. Alternatively, the amount administered to an infant is
from about 1 mg to about 10,000 mg/day, preferably abut 10 mg to
about 1000 mg, and yet more preferred about 10 mg to about 500
mg.
[0066] The protein source that is present in addition to the
subject dipeptide can be non-fat milk solids, a combination of
non-fat milk solids and whey protein, a partial hydrolysate of
non-fat milk and/or whey solids, soy protein isolates, or partially
hydrolyzed soy protein isolates. The infant formula can be casein
predominant or whey predominant.
[0067] The carbohydrate source in the infant formula can be any
suitable carbohydrate known in the art to be suitable for use in
infant formulas. Typical carbohydrate sources include sucrose,
fructose, glucose, maltodextrin, lactose, corn syrup, corn syrup
solids, rice syrup solids, rice starch, modified corn starch,
modified tapioca starch, rice flour, soy flour, and the like.
[0068] The lipid source in the infant formula can be any lipid or
fat known in the art to be suitable for use in infant formulas.
Typical lipid sources include milk fat, safflower oil, egg yolk
lipid, olive oil, coconut oil, palm oil, palm kernel oil, soybean
oil, sunflower oil, fish oil and fractions derived thereof such as
palm olein, medium chain triglycerides (MCT), and esters of fatty
acids wherein the fatty acids are, for example, arachidonic acid,
linoleic acid, palmitic acid, stearic acid, docosahexaenoic acid,
eicosapentaenoic acid, linolenic acid, oleic acid, lauric acid,
capric acid, caprylic acid, caproic acid, and the like. High oleic
forms of various oils are also contemplated to be useful herein
such as high oleic sunflower oil and high oleic safflower oil.
Medium chain triglycerides contain higher concentrations of
caprylic and capric acid than typically found in conventional oils,
e.g., approximately three-fourths of the total fatty acid content
is caprylic acid and one-fourth is capric acid.
[0069] Nutritionally complete compositions contain all vitamins and
minerals understood to be essential in the daily diet and these
should be present in nutritionally significant amounts. Those
skilled in the art appreciate that minimum requirements have been
established for certain vitamins and minerals that are known to be
necessary for normal physiological function. Practitioners also
understand that appropriate additional amounts (overages) of
vitamin and mineral ingredients need to be provided to compensate
for some loss during processing and storage of such
compositions.
[0070] To select a specific vitamin or mineral compound to be used
in the infant formula of the invention requires consideration of
that compound's chemical nature regarding compatibility with the
particular processing conditions used and shelf storage.
[0071] Examples of minerals, vitamins and other nutrients
optionally present in the composition of the invention include
vitamin A, vitamin B.sub.6, vitamin B.sub.12, vitamin E, vitamin K,
vitamin C, folic acid, thiamine, inositol, riboflavin, niacin,
biotin, pantothenic acid, choline, calcium, phosphorus, iodine,
iron, magnesium, copper, zinc, manganese, chloride, potassium,
sodium, selenium, chromium, molybdenum, taurine, and L-carnitine.
Minerals are usually added in salt form. In addition to
compatibility and stability considerations, the presence and
amounts of specific minerals and other vitamins will vary somewhat
depending on the intended infant population.
[0072] The infant formula of the invention also typically contains
emulsifiers and stabilizers such as soy lecithin, carrageenan, and
the like.
[0073] The infant formula of the invention may optionally contain
other substances which may have a beneficial effect such as
lactoferrin, nucleotides, nucleosides, immunoglobulins, and the
like.
[0074] The infant formula of the invention is in concentrate liquid
form, liquid ready to consume form, or powder form. Of course, if
in powder form, the formula is diluted to normal strength with
water to be in a form ready to consume.
[0075] The osmolality of the liquid infant formula of the invention
(when ready to consume) is typically about 100 to 1100 mOsm/kg
H.sub.2O, more typically about 200 to 700 mOsm/kg H.sub.2O.
[0076] The infant formula of the invention can be sterilized, if
desired, by techniques known in the art, for example, heat
treatment such as autoclaving or retorting, and the like.
[0077] The infant formula of the invention can be packaged in any
type of container known in the art to be used for storing
nutritional products such as glass, lined paperboard, plastic,
coated metal cans and the like.
[0078] The infant formula of the invention is shelf stable after
reconstitution. By "shelf stable" is meant that the formula in a
form ready to consume remains in a single homogenous phase (i.e.,
does not separate into more than one phase upon visual inspection)
or that the thickener does not settle out as a sediment upon visual
inspection after storage overnight in the refrigerator. With the
thickened nature of the product, the formula of the invention also
has the advantage of remaining fluid (i.e., does not gel into a
solid mass when stored overnight in the refrigerator).
[0079] In the method of the invention, infant formula comprising
arginyl-glutamine dipeptide, or a precursor thereof, is
administered to an infant. The form of administration is oral,
which includes tube feeding.
[0080] The invention provides a commercially acceptable product in
terms of desired stability and physical characteristics and the
product demonstrates little to no observable browning effect
by-products associated with a Maillard reaction.
[0081] Further, the inventive composition is substantially
homogeneous for an acceptable period after reconstitution (or for
the shelf-life if prepared as a liquid). The invention is
particularly useful for infant formula preparations for the
prevention and treatment of retionoathy of prematurity, although it
is equally applicable to other elemental diets specific to a
selected population that is at risk of, or is suspected of having,
diabetic retinopathy, vascular proliferative retinopathy, or
proliferation of abnormal vascularization, and the like.
[0082] One embodiment of the current invention envisions parenteral
administration, especially intravenous administration, as the route
of administration. Parenteral dosage forms should be sterile and
pyrogen-free, and are prepared in accord with accepted
pharmaceutical procedures. The parenteral formulations may be
organic or aqueous or mixed organic/aqueous formulations and may
further contain anti-oxidants, buffers, bacteriostats, isotonicity
adjusters and like additions acceptable for parenteral
formulations. In a preferred embodiment, the parenteral formulation
contains an effective amount of the peptide of the subject ivention
in an aqueous solution.
[0083] For parenteral application, particularly suitable are
injectable, sterile solutions, preferably oily or aqueous
solutions, as well as suspensions, emulsions, or implants. In
particular, carriers for parenteral administration include aqueous
solutions of dextrose, saline, pure water, ethanol, glycerol,
propylene glycol, peanut oil, sesame oil,
polyoxyethylene-polyoxypropylene block polymers, and the like.
Ampules are convenient unit dosages.
[0084] Also, according to the subject invention, the local
administration of the dipeptide compounds, and formulations
thereof, by means of a drug delivery device or implant placed in
proximity to the local tissue site provides for the maintenance of
efficacious, safe levels of active drug ingredient at the local
tissue disease site.
[0085] According to the subject invention, the local ocular
administration of dipeptide compounds of the invention, and/or
formulations thereof, attenuate ocular pathological disease
processes. Thus, local ocular administration of a dipeptide
compound of the invention, and/or formulations thereof, provides
for an efficacious but safe controlled concentration range of the
dipeptide directly in the eye.
[0086] Ocular dipeptide-based therapies, as describe herein,
provide significant advantages for treating neovascular ocular
disease relative to current laser surgery treatment modalities
including panretinal photocoagulation, which can be accompanied by
extensive ocular tissue damage. In the examples of posterior
neovascular ocular diseases, such as Age Related Macular
Degeneration and Diabetic Retinopathy, target ocular pathologies
and tissues for treatment are especially localized to the retinal,
choroidal and corneal ocular compartments.
[0087] Preferably, the peptide is administered locally to the eye,
retinal area, choroid area or associated vasculature. The peptide
can also be administered to the cornea of the eye. The peptide
diffuses into the eye and contacts the retina or surrounding
vasculature (e.g., eye drops, creams or gels).
[0088] The dipeptide compounds of the present invention, and
formulations thereof, are advantageous because they overcome
problems associated with stability, toxicity, lack of target tissue
specificity, safety, efficacy, extent and variability of
bioavailability.
[0089] As contemplated in the subject invention, where a dipeptide
compound comprises a prodrug, the prodrug can be converted to a
biologically active compound at a controlled rate via passive (such
as by aqueous hydrolysis) or biologically mediated (such as
biocatalytic or enzymatic) mechanisms. An advantage of the in vivo
conversion of the prodrug is that the ensuing dipeptide provides
localized therapeutic effects in target disease tissue with high
therapeutic margins of safety.
[0090] A further embodiment provides the use of dipeptide compounds
in conjunction with a drug delivery system in the form of an
implant or a device for the treatment of conditions as set forth
herein. Certain embodiments of the invention contemplate the use of
dipeptide compounds and formulations thereof for use as coatings
for example in conjunction with physical implants such as stents
and band ligatures. Therapeutic uses of such implants include but
are not limited to vascular diseases such as restenosis, and in
bone and tissue grafts.
[0091] A further embodiment of the subject invention provides for
the local administration of dipeptide compounds in combination with
other pharmacological therapies. As contemplated in the subject
invention, combination therapies of dipeptide compounds with other
medicaments targeting similar or distinct disease mechanisms have
advantages of greater efficacy and safety relative to respective
monotherapies with either specific medicament.
[0092] In one embodiment, a dipeptide compound is used to treat
neovascular ocular disease by localized (for example, in ocular
tissue) concurrent administration with other medicaments that act
to block angiogenesis by pharmacological mechanisms. Medicaments
that can be concurrently administered with a dipeptide compound of
the invention include, but are not limited to, vascular endothelial
growth factor VEGF blockers (e.g. by VEGF neutralizing binding
molecules such as Macugen (Eyetech) and Lucentis (ranibizumab,
Genentech), Squalamine lactate (Genaera Corporation); and VEGF
tyrosine kinase inhibition) for treating neovascular ocular disease
(AMD and Diabetic Retinopathy) and glucocorticoids (e.g.
Triamcinolone) for treating macular edema.
[0093] One or more active agents can be administered. When
administering more than one, the administration of the agents can
occur simultaneously or sequentially in time. The agents can be
administered before and after one another, or at the same time. The
methods also include co-administration with other drugs that are
used to treat retinopathy or other diseases described herein.
[0094] The composition can be administered in a single dose or in
more than one dose over a period of time to confer the desired
effect.
[0095] The dosage administration to a host in the above indications
will be dependent upon the specific condition being treated, the
type of host involved, its age, weight, health, kind of concurrent
treatment, if any, frequency of treatment, and therapeutic ratio.
Those skilled in the art will be able to determine the appropriate
dosages depending on these and other factors. Typically, a
therapeutically effective amount of the compound can range from
about 1 mg per day to about 1000 mg per day for an adult human
animal. For oral administration to human adults, a dosage of 0.01
to 100 mg/kg/day, preferably 0.01-1 mg/kg/day, is generally a
therapeutically effective amount.
[0096] To provide for the administration of such dosages for the
desired therapeutic treatment, new pharmaceutical compositions of
the invention will advantageously comprise between about 0.1% and
45%, and especially, 1 and 15%, by weight of the total of one or
more of the new compounds based on the weight of the total
composition including carrier or diluent.
[0097] In a retinal cell culture model used to study the effects of
the dipeptide on transepithelial resistance (TER) and vascular
endothelial growth factor (VEGF), it was demonstrated that the
dipeptide increased TER and decreased VEGF, both desirable effects
that have been associated with a decrease in vascular proliferative
retinal disease.
[0098] All patents, patent applications, provisional applications,
and publications referred to or cited herein are incorporated by
reference in their entirety to the extent they are not inconsistent
with the explicit teachings of this specification.
[0099] Following is an example which illustrates procedures for
practicing the invention. This example should not be construed as
limiting. All percentages are by weight and all solvent mixture
proportions are by volume unless otherwise noted.
EXAMPLE 1
Use of Arginyl-glutamine to Prevent Retinopathy of Prematurity
[0100] Neonatal mice were exposed to a period of hyperoxia to
induce retinal angiogenesis that mimics retinopathy of prematurity.
An Arginyl-glutamine dipeptide was administered intraperitoneally
twice daily during the period when neovascularization occurs in
this model, days 12 through 17.
[0101] On day 17 the animals are sacrificed and degree of
angiogenesis was quantified by counting pre-retinal
neovascularization on stained sections from treated and untreated
animals.
[0102] The data summarized in FIG. 1 represent combined data from
two separate experiments. The results demonstrated a statistically
significant effect of the dipeptide (argininyl-glutamine) compared
to vehicle and dipeptide (alaninyl-glycine). Treatment of the pups
with the dipeptide (argininyl-glutamine) resulted in a 80%
reduction in preretinal neovascularization.
[0103] This experiment has been repeated with different doses of
the dipeptide compared to the vehicle. The results are shown in
FIG. 2. This shows a clear decrease in nuclei (associated with
decreased retinopathy) in the animals receiving the dipeptide after
the oxygen stress challenge that is known to induce
retinopathy.
[0104] It should be understood that the examples and embodiments
described herein are for illustrative purposes only and that
various modifications or changes in light thereof will be suggested
to persons skilled in the art and are to be included within the
spirit and purview of this application.
* * * * *