U.S. patent application number 14/996800 was filed with the patent office on 2016-07-21 for methods and compositions for oral administration of proteins.
This patent application is currently assigned to Oramed Pharmaceuticals Inc.. The applicant listed for this patent is Oramed Pharmaceuticals Inc.. Invention is credited to Miriam Kidron.
Application Number | 20160206703 14/996800 |
Document ID | / |
Family ID | 37836252 |
Filed Date | 2016-07-21 |
United States Patent
Application |
20160206703 |
Kind Code |
A1 |
Kidron; Miriam |
July 21, 2016 |
METHODS AND COMPOSITIONS FOR ORAL ADMINISTRATION OF PROTEINS
Abstract
This invention provides compositions comprising a protein and an
omega-3 fatty acid, method for treating diabetes mellitus,
comprising administering same, and methods for oral administration
of a protein with an enzymatic activity, comprising orally
administering same.
Inventors: |
Kidron; Miriam; (Jerusalem,
IL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Oramed Pharmaceuticals Inc. |
Jerusalem |
|
IL |
|
|
Assignee: |
Oramed Pharmaceuticals Inc.
Jerusalem
IL
|
Family ID: |
37836252 |
Appl. No.: |
14/996800 |
Filed: |
January 15, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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11513343 |
Aug 31, 2006 |
9259456 |
|
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14996800 |
|
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60713716 |
Sep 6, 2005 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 9/4891 20130101;
A61K 31/202 20130101; A61P 43/00 20180101; A61K 45/06 20130101;
A61K 31/22 20130101; A61K 9/2866 20130101; A61K 9/2013 20130101;
A61K 47/28 20130101; A61K 35/60 20130101; A61K 38/56 20130101; A61K
9/0053 20130101; A61P 3/10 20180101; A61K 9/2846 20130101; A61K
9/4875 20130101; A61K 9/2068 20130101; A61K 38/28 20130101; A61K
9/5057 20130101; A61K 9/485 20130101; A61K 9/4866 20130101; A61K
38/56 20130101; A61K 2300/00 20130101; A61K 38/28 20130101; A61K
2300/00 20130101; A61K 35/60 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 38/28 20060101
A61K038/28; A61K 47/28 20060101 A61K047/28; A61K 45/06 20060101
A61K045/06; A61K 9/00 20060101 A61K009/00; A61K 31/202 20060101
A61K031/202; A61K 38/56 20060101 A61K038/56; A61K 47/18 20060101
A61K047/18; A61K 9/50 20060101 A61K009/50 |
Claims
1. A composition comprising a protein having a molecular weight of
up to 100,000 Daltons and an omega-3 fatty acid.
2. The composition of claim 1, wherein (a) said protein is insulin;
(b) said omega-3 fatty acid is derived from fish oil; or (c) said
inhibitor is soybean trypsin inhibitor (SBTI).
3. (canceled)
4. The composition of claim 1, further comprising an inhibitor of a
protease.
5. (canceled)
6. The composition of claim 4, wherein (a) said inhibitor is
AEBSF-HCI; (epsilon)-aminocaproic acid; (alpha)1-antichymotypsin;
antipain; antithrombin III; (alpha)1-antitrypsin
([alpha]1-proteinase inhibitor); APMSF-HCI (4-amidinophenyl-methane
sulfonyl-fluoride); sprotinin; benzamidine-HCI; chymostatin; DFP
(diisopropylfluoro-phosphate); leupeptin; PEFABLOC.RTM. SC
(4-(2-Aminoethyl)-benzenesulfonyl fluoride hydrochloride); PMSF
(phenylmethyl sulfonyl fluoride); TLCK
(1-Chloro-3-tosylamido-7-amino-2-beptanone HCI); TPCK
(1-Chloro-3-tosylarnido-4-phenyl-2-butanone); trypsin inhibitor
from egg white (Ovomucoid); trypsin inhibitor from soybean;
aprotinin; pentamidine isethionate; pepstatin; guanidium;
alpha2-macroglobulin; a chelating agent of zinc; iodoacetate; or
zinc; (b) said protease is a serine protease; or (c) said protease
is trypsin.
7-8. (canceled)
9. The composition of claim 1, further comprising a substance that
enhances absorption of said insulin protein through an intestinal
mucosal barrier.
10. The composition of claim 9, wherein said substance is EDTA
11. The composition of claim 9, wherein said substance is a bile
acid or alkali metal salt thereof.
12. The composition of claim 11, wherein said bile acid is cholic
acid, chenodeoxycholic acid, taurocholic acid,
taurochenodeoxycholic acid, glycocholic acid, glycochenocholic
acid, 3.beta.-monohydroxychloric acid, lithocholic acid,
3.alpha.-hydroxy-12-ketocholic acid, 3.beta.-hydroxy-12-ketocholic
acid, 12.alpha.-3.beta.-dihydrocholic acid, or ursodesoxycholic
acid.
13. The composition of claim 1, further comprising a coating that
inhibits digestion of said composition in a stomach of a
subject.
14. The composition of claim 13, wherein said coating is an enteric
coating or gelatin coating.
15. A method for oral administration of a protein having a
molecular weight up to 100,000 Daltons to a subject, whereby a
substantial fraction of said protein retains its activity after
absorption, through an intestinal mucosal barrier of said subject,
comprising administering orally to said subject a pharmaceutical
composition comprising said protein and an omega-3 fatty acid.
16. The method of claim 15, wherein (a) said protein is an enzyme;
(b) said protein is insulin; (c) said protein is a glucagon, an
interferon gamma, an interferon alpha, a growth hormone, an
erythropoietin, or granulocyte colony stimulating factor (G-CSF);
(d) said protein has a molecular weight of 1-50 kilodalton; (e)
said protein is a receptor ligand, transport protein, storage
protein or a combination thereof; or (f) said composition further
comprises omega-3 fatty acid derived from fish oil.
17-21. (canceled)
22. The method of claim 15, wherein said pharmaceutical composition
further comprises a protease inhibitor.
23. The method of claim 22, wherein (a) said inhibitor is soybean
trypsin inhibitor (SBTI); (b) said inhibitor is AEBSF-HCI;
(epsilon)-aminocaproic acid; (alpha)1-antichymotypsin; antipain;
antithrombin III; (alpha)1-antitrypsin ([alpha]1-proteinase
inhibitor); APMSF-HCI (4-amidinophenyl-methane sulfonyl-fluoride);
sprotinin; benzamidine-HCI; chymostatin; DFP
(diisopropylfluoro-phosphate); leupeptin; PEFABLOC.RTM. SC
(4-(2-Aminoethyl)-benzenesulfonyl fluoride hydrochloride); PMSF
(phenylmethyl sulfonyl fluoride); TLCK
(1-Chloro-3-tosylamido-7-amino-2-heptanone HCI); TPCK
(1-Chloro-3-tosylamido-4-phenyl-2-butanone); trypsin inhibitor from
egg white (Ovomucoid); trypsin inhibitor from soybean; aprotinin;
pentamidine isethionate; pepstatin; guanidium;
alpha2-macroglobulin; a chelating agent of zinc; iodoacetate; or
zinc; (c) wherein said protease is a serine protease; or (d)
wherein said protease is trypsin.
24-26. (canceled)
27. The method of claim 5, wherein said pharmaceutical composition
further comprises a substance that enhances absorption of said
protein through an intestinal mucosal barrier.
28. The method of claim 27, wherein said substance is EDTA.
29. The method of claim 27, wherein said substance is a bile acid
or alkali metal salt thereof.
30. The method of claim 29, wherein said bile acid is cholic acid,
chenodeoxycholic acid, taurocholic acid, taurochenodeoxycholic
acid, glycocholic acid, glycochenocholic acid,
3.beta.-monohydroxychloric acid, lithocholic acid,
3.alpha.-hydroxy-12-ketocholic acid, 3.beta.-hydroxy-12-ketocholic
acid, 12.alpha.-3.beta.-dihydrocholic acid, or ursodesoxycholic
acid.
31. The method of claim 15, wherein said pharmaceutical composition
further comprises a coating that inhibits digestion of said
composition in a stomach of a subject.
32. The method of claim 31, wherein said coating is an enteric
coating or gelatin coating.
33. A method for treating diabetes mellitus in a subject,
comprising administering orally to said subject a pharmaceutical
composition comprising insulin and an omega-3 fatty acid, thereby
treating diabetes mellitus.
34. The method of claim 33, wherein (a) said composition further
comprises omega-3 fatty acid derived from fish oil; or (b) said
pharmaceutical composition further comprises a coating that
inhibits digestion of said composition in a stomach of a
subject.
35. The method of claim 33, wherein said pharmaceutical composition
further comprises an inhibitor of a protease.
36. The method of claim 35, wherein (a) said inhibitor is soybean
trypsin inhibitor (SBTI); (b) said inhibitor is AEBSF-HCI;
(epsilon)-aminocaproic acid; (alpha)1-antichymotypsin; antipain;
antithrombin III; (alpha)1-antitrypsin ([alpha]1-proteinase
inhibitor); APMSF-HCI (4-ainidinophenyl-methane sulfonyl-fluoride);
sprotinin; benzamidine-HCI; chymostatin; DFP
(diisopropylfluoro-phosphate); leupeptin; PEFABLOC.RTM. SC (4-(2
Aminoethyl)-benzenesulfonyl fluoride hydrochloride); PMSF
(phenylmethyl sulfonyl fluoride); TLCK
(1-Chloro-3-tosylamido-7-amino-2-heptanone HCI); TPCK
(1-Chloro-3-tosylamido-4-phenyl-2-butanone); trypsin inhibitor from
egg white (Ovomucoid); trypsin inhibitor from soybean; aprotinin;
pentamidine isethionate; pepstatin; guanidium;
alpha2-macroglobulin; a chelating agent of zinc; iodoacetate; or
zinc; (c) said protease is a serine protease; or (d) said protease
is trypsin.
37-39. (canceled)
40. The method of claim 33, wherein said pharmaceutical composition
further comprises a substance that enhances absorption of said
insulin protein through an intestinal mucosal barrier.
41. The method of claim 40, wherein said substance is EDTA.
42. The method of claim 40, wherein said substance is a bile acid
or alkali metal salt thereof.
43. The method of claim 42, wherein said bile acid is cholic acid,
chenodeoxycholic acid, taurncholic acid, taurochenodeoxycholic
acid, glycocholic acid, glycochenocholic acid,
3.beta.-monohydroxychloric acid, lithocholic acid,
3.alpha.-hydroxy-12-ketocholic acid, 3.beta.-hydroxy-12-ketocholic
acid, 12.alpha.-3.beta.-dihydrocholic acid, or ursodesoxycholic
acid.
44. (canceled)
45. The method of claim 41, wherein said coating is an enteric
coating or gelatin coating.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority of U.S. Provisional Ser.
No. 60/713,716, filed on Sep. 6, 2005, which is included in its
entirety by reference herein.
FIELD OF INVENTION
[0002] This invention provides compositions comprising a protein
and an omega-3 fatty acid, and a method for administering same.
BACKGROUND OF THE INVENTION
[0003] Due to improved biotechnology, the accessibility of
biologically active peptides to the pharmaceutical industry has
increased considerably. However, a limiting factor in the
development of peptide drugs is the relative ineffectiveness when
given perorally. Almost all peptide drugs are parenterally
administered, although parenterally administered peptide drugs are
often connected with low patient compliance.
[0004] Insulin is a medicament used to treat patients suffering
from diabetes, and is the only treatment for insulin-dependent
diabetes mellitus. Diabetes Mellitus is characterized by a
pathological condition of absolute or relative insulin deficiency,
leading to hyperglycemia, and is one of the main threats to human
health in the 21st century. The global figure of people with
diabetes is set to rise to 220 million in 2010, and 300 million in
2025. Type I diabetes is caused primarily by the failure of the
pancreas to produce insulin. Type II diabetes, involves a lack of
responsiveness of the body to the action of insulin.
[0005] Approximately 20%-30% of all diabetics use daily insulin
injections to maintain their glucose levels. An estimated 10% of
all diabetics are totally dependent on insulin injections.
[0006] Currently, the only route of insulin administration is
injection. Daily injection of insulin is causes considerable
suffering for patients. Side effects such as lipodystrophy at the
site of the injection, lipatrophy, lilpohypertrophy, and occasional
hypoglycemia are known to occur. In addition, subcutaneous
administration of insulin does not typically provide the fine
continuous regulation of metabolism that occurs normally with
insulin secreted from the pancreas directly into the liver via the
portal vein.
[0007] The present invention addresses the need for an alternate
solution for administration of insulin.
SUMMARY OF THE INVENTION
[0008] This invention provides compositions comprising a protein
and an omega-3 fatty acid, method for treating diabetes mellitus,
comprising administering same, and methods for oral administration
of a protein with an enzymatic activity, comprising orally
administering same.
[0009] In one embodiment, the present invention provides a
composition comprising an insulin protein and an omega-3 fatty
acid.
[0010] In another embodiment, the present invention provides a
method for oral administration of a protein with an enzymatic
activity to a subject, whereby a substantial fraction of the
protein retains the enzymatic activity after absorption through an
intestinal mucosal barrier of the subject, comprising administering
orally to the subject a pharmaceutical composition comprising the
protein and an omega-3 fatty acid, thereby orally administering a
protein with an enzymatic activity to a subject.
[0011] In another embodiment, the present invention provides a
method for treating diabetes mellitus in a subject, comprising
administering orally to the subject a pharmaceutical composition
comprising insulin and an omega-3 fatty acid, thereby treating
diabetes mellitus.
DETAILED DESCRIPTION OF THE INVENTION
[0012] This invention provides compositions and methods comprising
a protein and an omega-3 fatty acid. In one embodiment, the protein
having a molecular weight up to 200,000 Daltons. In a preferred
embodiment, the protein having a molecular weight up to 100,000
Daltons. In one embodiment, the present invention further provides
an enhancer which enhances absorption through the intestines.
[0013] In one embodiment, the protein is an enzyme. In some
embodiments, the protein is a receptor ligand, transporter, or a
storage protein. In one embodiment, the protein is a structural
protein.
[0014] In some embodiments, the enzyme is an oxidoreductase,
transferase, hydrolase, lyase, isomerase, or ligase. In some
embodiments, oxidoreductases act on the aldehyde or oxo group of
donors, on the CH--CH group of donors, on the CH--NH(2) group of
donors, on the CH--NH group of donors, on NADH or NADPH, on the
CH--OH group of donors, on nitrogenous compounds as donors, on a
sulfur group of donors, on a heme group of donors, on diphenols and
related substances as donors, on a peroxide as acceptor, on
hydrogen as donor, on single donors with incorporation of molecular
oxygen, on paired donors, on superoxide as acceptor, oxidizing
metal ions, on CH or CH(2) groups, on iron-sulfur proteins as
donors, on reduced flavodoxin as donor, on phosphorus or arsenic in
donors, or on x-H and y-H to form an x-y bond.
[0015] In some embodiments, transferases are acyltransferases or
glycosyltransferases. In some embodiments, transferases transfer
aldehyde or ketone residues. In some embodiments, transferases
transfer alkyl or aryl groups, other than methyl groups. In some
embodiments, transferases transfer nitrogenous, phosphorous, sulfur
or selenium containing groups.
[0016] In some embodiments, hydrolases are glycosylases or act on
ether bonds, on peptide bonds, on carbon-nitrogen bonds, other than
peptide bonds, on acid anhydrides, on carbon-carbon bonds, on
halide bonds, on phosphorus-nitrogen bonds, on sulfur-nitrogen
bonds, on carbon-phosphorus bonds, on sulfur-sulfur bonds, or on
carbon-sulfur bonds.
[0017] In some embodiments, lyases are carbon-carbon lyases,
carbon-oxygen lyases, carbon-nitrogen lyases, carbon-sulfur lyases,
carbon-halide lyases, phosphorus-oxygen lyases, or other
lyases.
[0018] In some embodiments, isomerases are racemases or epimerases,
cis-trans-isomerases, intramolecular oxidoreductases,
intramolecular transferases, intramolecular lyases, or other
isomerases.
[0019] In some embodiments, ligases form carbon-sulfur bonds,
carbon-nitrogen bonds, carbon-carbon bonds, phosphoric ester bonds,
or nitrogen-metal bonds.
[0020] In some embodiments, transporter proteins are annexins,
ATP-binding cassette transporters, hemoglobin, ATPases, calcium
channels, potassium channels, sodium channels, or solute
carriers.
[0021] In some embodiments, storage proteins comprise albumins,
lactoglobulins, casein ovomucin, ferritin, phosvitin, lactoferrin,
or vitellogenin. In one embodiment, albumins comprise avidin,
ovalbumin, serum albumin, parvalbumin, c-reactive protein
prealbumin, conalbumin, ricin, lactalbumin, methemalbumin, or
transthyretin.
[0022] In some embodiments, structural proteins comprise amyloid,
collagen elastin, or fibrillin.
[0023] In some embodiments, the protein is a viral protein,
bacterial protein, invertebrate protein, or vertebrate protein. In
some embodiments, the protein is a recombinant protein. In one
embodiment, the protein is a recombinant protein. In one
embodiment, the recombinant protein is a recombinant human
protein.
[0024] In one embodiment, the present invention provides a
composition comprising an insulin protein and an omega-3 fatty
acid. As provided herein (Examples), such compositions have utility
in the oral administration of insulin, whereby the insulin is
absorbed by the intestines into the bloodstream in an active
form.
[0025] In another embodiment, the present invention provides a
composition comprising a protein with enzymatic activity and an
omega-3 fatty acid.
[0026] In one embodiment, the insulin of methods and compositions
of the present invention is human insulin. In another embodiment,
the insulin is a recombinant insulin. In another embodiment, the
insulin is recombinant human insulin. In another embodiment, the
insulin is bovine insulin. In another embodiment, the insulin is
porcine insulin. In another embodiment, the insulin is whale
insulin. In another embodiment, the insulin is a metal complex of
insulin (e.g. a zinc complex of insulin, protamine zinc insulin, or
globin zinc).
[0027] In another embodiment, the insulin is regular insulin. In
another embodiment, the insulin is fast-acting insulin. In another
embodiment, the insulin is lente insulin. In another embodiment,
the insulin is semilente insulin. In another embodiment, the
insulin is Ultralente insulin. In another embodiment, the insulin
is NPH insulin. In another embodiment, the insulin is glargine
insulin. In another embodiment, the insulin is lispro insulin. In
another embodiment, the insulin is aspart insulin. In another
embodiment, the insulin is a combination of two or more of any of
the above types of insulin. In another embodiment, the insulin is
any other type of insulin known in the art. Each possibility
represents a separate embodiment of the present invention.
[0028] In one embodiment, the amount of insulin utilized in methods
and compositions of the present invention is 0.5-3 units (u)/kg in
humans. In one embodiment, the units used to measure insulin in
methods and compositions of the present invention are USP Insulin
Units. In one embodiment, the units used to measure insulin are
milligrams. In another embodiment, one USP Insulin Unit is
equivalent to 45.5 mg insulin.
[0029] In another embodiment, the amount of insulin is 0.1-1 u/kg.
In another embodiment, the amount is 0.2-1 u/kg. In another
embodiment, the amount is 0.3-1 u/kg. In another embodiment, the
amount is 0.5-1 u/kg. In another embodiment, the amount is 0.1-2
u/kg. In another embodiment, the amount is 0.2-2 u/kg.
[0030] In another embodiment, the amount is 0.3-2 u/kg. In another
embodiment, the amount is 0.5-2 u/kg. In another embodiment, the
amount is 0.7-2 u/kg. In another embodiment, the amount is 1-2
u/kg. In another embodiment, the amount is 1.2-2 u/kg. In another
embodiment, the amount is 1-1.2 u/kg. In another embodiment, the
amount is 1-1.5 u/kg. In another embodiment, the amount is 1-2.5
u/kg. In another embodiment, the amount is 1-3 u/kg. In another
embodiment, the amount is 2-3 u/kg. In another embodiment, the
amount is 1-5 u/kg. In another embodiment, the amount is 2-5 u/kg.
In another embodiment, the amount is 3-5 u/kg.
[0031] In another embodiment, the amount of insulin is 0.1 u/kg. In
another embodiment, the amount is 0.2 u/kg. In another embodiment,
the amount is 0.3 u/kg. In another embodiment, the amount is 0.4
u/kg. In another embodiment, the amount is 0.5 u/kg. In another
embodiment, the amount is 0.6 u/kg. In another embodiment, the
amount is 0.8 u/kg. In another embodiment, the amount is 1 u/kg. In
another embodiment, the amount is 1.2 u/kg. In another embodiment,
the amount is 1.4 u/kg. In another embodiment, the amount is 1.6
u/kg. In another embodiment, the amount is 1.8 u/kg. In another
embodiment, the amount is 2 u/kg. In another embodiment, the amount
is 2.2 u/kg. In another embodiment, the amount is 2.5 u/kg. In
another embodiment, the amount is 3 u/kg.
[0032] In another embodiment, the amount of insulin is 1-10 u. In
another embodiment, the amount is 2-10 u. In another embodiment,
the amount is 3-10 u. In another embodiment, the amount is 5-10 u.
In another embodiment, the amount is 1-20 u. In another embodiment,
the amount is 2-20 u. In another embodiment, the amount is 3-20 u.
In another embodiment, the amount is 5-20 u. In another embodiment,
the amount is 7-20 u. In another embodiment, the amount is 10-20 u.
In another embodiment, the amount is 12-20 u. In another
embodiment, the amount is 10-12 u. In another embodiment, the
amount is 10-15 u. In another embodiment, the amount is 10-25 u. In
another embodiment, the amount is 10-30 u. In another embodiment,
the amount is 20-30 u. In another embodiment, the amount is 10-50
u. In another embodiment, the amount is 20-50 u. In another
embodiment, the amount is 30-50 u. In another embodiment, the
amount is 20-100 u. In another embodiment, the amount is 30-100 u.
In another embodiment, the amount is 100-150 u. In another
embodiment, the amount is 100-250 u. In another embodiment, the
amount is 100-300 u. In another embodiment, the amount is 200-300
u. In another embodiment, the amount is 100-500 u. In another
embodiment, the amount is 200-500 u. In another embodiment, the
amount is 300-500 u. In another embodiment, the amount is 200-1000
u. In another embodiment, the amount is 300-1000 u.
[0033] In another embodiment, the amount of insulin is 1 u. In
another embodiment, the amount is 2 u. In another embodiment, the
amount is 3 u. In another embodiment, the amount is 4 u. In another
embodiment, the amount is 5 u. In another embodiment, the amount is
6 u. In another embodiment, the amount is 8 u. In another
embodiment, the amount is 10 u. In another embodiment, the amount
is 12 u. In another embodiment, the amount is 14 u. In another
embodiment, the amount is 16 u. In another embodiment, the amount
is 18 u. In another embodiment, the amount is 20 u. In another
embodiment, the amount is 22 u. In another embodiment, the amount
is 25 u. In another embodiment, the amount is 30 u. In another
embodiment, the amount is 50 u. In another embodiment, the amount
is 80 u. In another embodiment, the amount is 100 u. In another
embodiment, the amount is 120 u. In another embodiment, the amount
is 140 u. In another embodiment, the amount is 160 u. In another
embodiment, the amount is 180 u. In another embodiment, the amount
is 200 u. In another embodiment, the amount is 300 u. In another
embodiment, the amount is 500 u.
[0034] In another embodiment, the use of sustained release dosage
forms (e.g. sustained release microencapsulation) enables the
treatment frequency to be reduced to once or twice a day. In
another embodiment, the insulin dosage is increased correspondingly
with decreasing frequency of administration.
[0035] Each amount of insulin represents a separate embodiment of
the present invention.
[0036] Methods of measuring insulin levels are well known in the
art. In one embodiment, levels of recombinant insulin are measuring
using a human insulin radio-immunoassay (RIA) kit, e.g. the kit
manufactured by Linco Research Inc, (St. Charles, Mo.). In another
embodiment, levels of C peptide are measured as well, to determine
the relative contributions of endogenous and exogenous insulin to
observed rises in insulin levels. In another embodiment, insulin
ELISA kits are used. In another embodiment, insulin levels are
measured by any other method known in the art. Each possibility
represents a separate embodiment of the present invention.
[0037] In some embodiments, omega-3 fatty acid can be found in
vegetable sources such as the seeds of chia, perilla, flax,
walnuts, purslane, lingonberry, seabuckthorn, and hemp. In some
embodiments, omega-3 fatty acids can also be found in the fruit of
the acai palm. In another embodiment, the omega-3 fatty acid has
been provided in the form of a synthetic omega-3 fatty acid. In one
embodiment, the omega-3 fatty acid of methods and compositions of
the present invention has been provided to the composition in the
form of a fish oil. In another embodiment, the omega-3 fatty acid
has been provided in the form of canola oil. In another embodiment,
the omega-3 fatty acid has been provided in the form of flaxseed
oil. In another embodiment, the omega-3 fatty acid has been
provided in the form of any other omega-3 fatty acid-rich source
known in the art. In another embodiment, the omega-3 fatty acid has
been provided in the form of a synthetic omega-3 fatty acid. Each
form of omega-3 fatty acids represents a separate embodiment of the
present invention.
[0038] In another embodiment, the omega-3 fatty acid of methods and
compositions of the present invention is an omega-3 polyunsaturated
fatty acid. In another embodiment, the omega-3 fatty acid is DHA,
an omega-3, polyunsaturated, 22-carbon fatty acid also referred to
as 4, 7, 10, 13, 16, 19-docosahexaenoic acid. In another
embodiment, the omega-3 fatty acid is a-linolenic acid (9, 12,
15-octadecatrienoic acid). In another embodiment, the omega-3 fatty
acid is stearidonic acid (6, 9, 12, 15-octadecatetraenoic acid). In
another embodiment, the omega-3 fatty acid is eicosatrienoic acid
(ETA; 11, 14, 17-eicosatrienoic acid). In another embodiment, the
omega-3 fatty acid is eicsoatetraenoic acid (8, 11, 14,
17-eicosatetraenoic acid). In one embodiment, the omega-3 fatty
acid is eicosapentaenoic acid (EPA; 5, 8, 11, 14,
17-eicosapentaenoic acid). In another embodiment, the omega-3 fatty
acid is eicosahexaenoic acid (also referred to as "EPA"; 5, 7, 9,
11, 14, 17-eicosahexaenoic acid). In another embodiment, the
omega-3 fatty acid is docosapentaenoic acid (DPA; 7, 10, 13, 16,
19-docosapenatenoic acid). In another embodiment, the omega-3 fatty
acid is tetracosahexaenoic acid (6, 9, 12, 15, 18,
21-tetracosahexaenoic acid). In another embodiment, the omega-3
fatty acid is any other omega-3 fatty acid known in the art. Each
omega-3 fatty acid represents a separate embodiment of the present
invention.
[0039] In another embodiment, compositions of the present invention
further comprise an inhibitor of a protease. As provided herein,
protease inhibitors enhance the ability of omega-3 fatty acids to
protect insulin and facilitate its absorption in the intestine.
[0040] In some embodiments, protease inhibitor inhibits the
function of peptidases. In one embodiment, protease inhibitors
enhance the ability of omega-3 fatty acids to protect the protein
of the present invention and facilitate its absorption in the
intestine. In some embodiments, the protease inhibitor of the
present invention is a protein. In some embodiments, protease
inhibitors comprise cysteine protease inhibitors, serine protease
inhibitors (serpins), trypsin inhibitors, threonine protease
inhibitors, aspartic protease inhibitors, metallo protease
inhibitors. In some embodiments, protease inhibitors comprise
suicide inhibitor, transition state inhibitor, or chelating
agents.
[0041] In one embodiment, the protease inhibitor is soybean trypsin
inhibitor (SBTI). In another embodiment, the protease inhibitor is
AEBSF-HCL. In another embodiment, the inhibitor is
(epsilon)-aminocaproic acid. In another embodiment, the inhibitor
is (alpha) 1-antichymotypsin. In another embodiment, the inhibitor
is antipain. In another embodiment, the inhibitor is antithrombin
III. In another embodiment, the inhibitor is (alpha) 1-antitrypsin
([alpha] 1-proteinase inhibitor). In another embodiment, the
inhibitor is APMSF-HCI (4-amidinophenyl-methane sulfonyl-fluoride).
In another embodiment, the inhibitor is sprotinin. In another
embodiment, the inhibitor is benzamidine-HCI. In another
embodiment, the inhibitor is chymostatin. In another embodiment,
the inhibitor is DFP (diisopropylfluoro-phosphate). In another
embodiment, the inhibitor is leupeptin. In another embodiment, the
inhibitor is PEFABLOC.RTM. SC (4-(2-Aminoethyl)-benzenesulfonyl
fluoride hydrochloride). In another embodiment, the inhibitor is
PMSF (phenylmethyl sulfonyl fluoride). In another embodiment, the
inhibitor is TLCK (1-Chloro-3-tosylamido-7-amino-2-heptanone HO).
In another embodiment, the inhibitor is TPCK
(1-Chloro-3-tosylamido-4-phenyl-2-butanone). In another embodiment,
the inhibitor is trypsin inhibitor from egg white (Ovomucoid). In
another embodiment, the inhibitor is trypsin inhibitor from
soybean. In another embodiment, the inhibitor is aprotinin. In
another embodiment, the inhibitor is pentamidine isethionate. In
another embodiment, the inhibitor is pepstatin. In another
embodiment, the inhibitor is guanidium. In another embodiment, the
inhibitor is alpha2-macroglobulin. In another embodiment, the
inhibitor is a chelating agent of zinc. In another embodiment, the
inhibitor is iodoacetate. In another embodiment, the inhibitor is
zinc. Each possibility represents a separate embodiment of the
present invention.
[0042] In another embodiment, the amount of protease inhibitor
utilized in methods and compositions of the present invention is
0.1 mg/dosage unit. In another embodiment, the amount of protease
inhibitor is 0.2 mg/dosage unit. In another embodiment, the amount
is 0.3 mg/dosage unit. In another embodiment, the amount is 0.4
mg/dosage unit. In another embodiment, the amount is 0.6 mg/dosage
unit. In another embodiment, the amount is 0.8 mg/dosage unit. In
another embodiment, the amount is 1 mg/dosage unit. In another
embodiment, the amount is 1.5 mg/dosage unit. In another
embodiment, the amount is 2 mg/dosage unit. In another embodiment,
the amount is 2.5 mg/dosage unit. In another embodiment, the amount
is 3 mg/dosage unit. In another embodiment, the amount is 5
mg/dosage unit. In another embodiment, the amount is 7 mg/dosage
unit. In another embodiment, the amount is 10 mg/dosage unit. In
another embodiment, the amount is 12 mg/dosage unit. In another
embodiment, the amount is 15 mg/dosage unit. In another embodiment,
the amount is 20 mg/dosage unit. In another embodiment, the amount
is 30 mg/dosage unit. In another embodiment, the amount is 50
mg/dosage unit. In another embodiment, the amount is 70 mg/dosage
unit. In another embodiment, the amount is 100 mg/dosage unit.
[0043] In another embodiment, the amount of protease inhibitor is
0.1-1 mg/dosage unit. In another embodiment, the amount of protease
inhibitor is 0.2-1 mg/dosage unit. In another embodiment, the
amount is 0.3-1 mg/dosage unit. In another embodiment, the amount
is 0.5-1 mg/dosage unit. In another embodiment, the amount is 0.1-2
mg/dosage unit. In another embodiment, the amount is 0.2-2
mg/dosage unit. In another embodiment, the amount is 0.3-2
mg/dosage unit. In another embodiment, the amount is 0.5-2
mg/dosage unit. In another embodiment, the amount is 1-2 mg/dosage
unit. In another embodiment, the amount is 1-10 mg/dosage unit. In
another embodiment, the amount is 2-10 mg/dosage unit. In another
embodiment, the amount is 3-10 mg/dosage unit. In another
embodiment, the amount is 5-10 mg/dosage unit. In another
embodiment, the amount is 1-20 mg/dosage unit. In another
embodiment, the amount is 2-20 mg/dosage unit. In another
embodiment, the amount is 3-20 mg/dosage unit. In another
embodiment, the amount is 5-20 mg/dosage unit. In another
embodiment, the amount is 10-20 mg/dosage unit. In another
embodiment, the amount is 10-100 mg/dosage unit. In another
embodiment, the amount is 20-100 mg/dosage unit. In another
embodiment, the amount is 30-100 mg/dosage unit. In another
embodiment, the amount is 50-100 mg/dosage unit. In another
embodiment, the amount is 10-200 mg/dosage unit. In another
embodiment, the amount is 20-200 mg/dosage unit. In another
embodiment, the amount is 30-200 mg/dosage unit. In another
embodiment, the amount is 50-200 mg/dosage unit. In another
embodiment, the amount is 100-200 mg/dosage unit.
[0044] In another embodiment, the amount of protease inhibitor
utilized in methods and compositions of the present invention is
1000 k.i.u. (kallikrein inactivator units)/pill. In another
embodiment, the amount is 10 k.i.u./dosage unit. In another
embodiment, the amount is 12 k.i.u./dosage unit. In another
embodiment, the amount is 15 k.i.u./dosage unit. In another
embodiment, the amount is 20 k.i.u./dosage unit. In another
embodiment, the amount is 30 k.i.u./dosage unit. In another
embodiment, the amount is 40 k.i.u./dosage unit. In another
embodiment, the amount is 50 k.i.u./dosage unit. In another
embodiment, the amount is 70 k.i.u./dosage unit. In another
embodiment, the amount is 100 k.i.u./dosage unit. In another
embodiment, the amount is 150 k.i.u./dosage unit. In another
embodiment, the amount is 200 k.i.u./dosage unit. In another
embodiment, the amount is 300 k.i.u./dosage unit. In another
embodiment, the amount is 500 k.i.u./dosage unit. In another
embodiment, the amount is 700 k.i.u./dosage unit. In another
embodiment, the amount is 1500 k.i.u./dosage unit. In another
embodiment, the amount is 3000 k.i.u./dosage unit. In another
embodiment, the amount is 4000 k.i.u./dosage unit. In another
embodiment, the amount is 5000 k.i.u./dosage unit.
[0045] Each amount of protease inhibitor represents a separate
embodiment of the present invention.
[0046] In another embodiment, the protease targeted by the protease
inhibitor of methods and compositions of the present invention is a
serine protease. In another embodiment, the protease is trypsin. In
another embodiment, the protease is chymotrypsin. In another
embodiment, the protease is carboxypeptidase. In another
embodiment, the protease is aminopeptidase. In another embodiment,
the protease is any other protease that functions in the duodenum
or the small intestine. Each possibility represents a separate
embodiment of the present invention.
[0047] In another embodiment, compositions of the present invention
further comprise a substance that enhances absorption of the
insulin through an intestinal mucosal barrier. Such a substance is
referred to herein as an "enhancer." As provided herein, enhancers,
when used together with omega-3 fatty acids, enhance the ability of
insulin to be absorbed in the intestine.
[0048] In one embodiment, the enhancer is
didecanoylphosphatidylcholine (DDPC). In one embodiment, the
enhancer is a chelating agent such as ethylenediaminetetraacetic
acid (EDTA) or egtazic acid EGTA. In a preferred embodiment, EDTA
is sodium-EDTA. In some embodiments, the enhancer is NO donor. In
some embodiments, the enhancer is a bile acid, glycine-conjugated
form of a bile acid, or an alkali metal salt. In one embodiment,
absorption enhancement is achieved through utilization of a
combination of .alpha.-galactosidase and .beta.-mannanase. In some
embodiments, the enhancer is a fatty acid such as sodium caprate.
In one embodiment, the enhancer is sodium glycocholate. In one
embodiment, the enhancer is sodium salicylate. In one embodiment,
the enhancer is n-dodecyl-.beta.-D-maltopyranoside.
[0049] In some embodiments, surfactants serve as absorption
enhancer. In one embodiment, the enhancer is chitisan such as
N,N,N-trimethyl chitosan chloride (TMC).
[0050] In one embodiment, NO donors of the present invention
comprise
3-(2-Hydroxy-1-(1-methylethyl)-2-nitrosohydrazino)-1-propanamine,
N-ethyl-2-(1-ethyl-hydroxy-2-nitrosohydrazino)-ethanamine, or
S-Nitroso-N-acetylpenicillamine
[0051] In another embodiment, the bile acid is cholic acid. In
another embodiment, the bile acid is chenodeoxycholic acid. In
another embodiment, the bile acid is taurocholic acid. In another
embodiment, the bile acid is taurochenodeoxycholic acid. In another
embodiment, the bile acid is glycocholic acid. In another
embodiment, the bile acid is glycochenocholic acid. In another
embodiment, the bile acid is 3 beta-monohydroxychloric acid. In
another embodiment, the bile acid is lithocholic acid. In another
embodiment, the bile acid is 5 beta-cholanic acid. In another
embodiment, the bile acid is 3,12-diol-7-one-5 beta-cholanic acid.
In another embodiment, the bile acid is 3
alpha-hydroxy-12-ketocholic acid. In another embodiment, the bile
acid is 3 beta-hydroxy-12-ketocholic acid. In another embodiment,
the bile acid is 12 alpha-3 beta-dihydrocholic acid. In another
embodiment, the bile acid is ursodesoxycholic acid.
[0052] In one embodiment, the enhancer is a nonionic surfactant. In
one embodiment, the enhancer is a nonionic polyoxyethylene ether
surface active agent (e.g one having an HLB value of 6 to 19,
wherein the average number of polyoxyethylene units is 4 to 30). In
another embodiment, the enhancer is an anionic surface active
agents. In another embodiment, the enhancer is a cationic surface
active agent. In another embodiment, the enhancer is an ampholytic
surface active agent. In one embodiment, zwitteruionic surfactants
such as acylcarnitines serve as absorption enhancers.
[0053] In another embodiment, the amount of enhancer utilized in
methods and compositions of the present invention is 0.1 mg/dosage
unit. In another embodiment, the amount of enhancer is 0.2
mg/dosage unit. In another embodiment, the amount is 0.3 mg/dosage
unit. In another embodiment, the amount is 0.4 mg/dosage unit. In
another embodiment, the amount is 0.6 mg/dosage unit. In another
embodiment, the amount is 0.8 mg/dosage unit. In another
embodiment, the amount is 1 mg/dosage unit. In another embodiment,
the amount is 1.5 mg/dosage unit. In another embodiment, the amount
is 2 mg/dosage unit. In another embodiment, the amount is 2.5
mg/dosage unit. In another embodiment, the amount is 3 mg/dosage
unit. In another embodiment, the amount is 5 mg/dosage unit. In
another embodiment, the amount is 7 mg/dosage unit. In another
embodiment, the amount is 10 mg/dosage unit. In another embodiment,
the amount is 12 mg/dosage unit. In another embodiment, the amount
is 15 mg/dosage unit. In another embodiment, the amount is 20
mg/dosage unit. In another embodiment, the amount is 30 mg/dosage
unit. In another embodiment, the amount is 50 mg/dosage unit. In
another embodiment, the amount is 70 mg/dosage unit. In another
embodiment, the amount is 100 mg/dosage unit.
[0054] In another embodiment, the amount of enhancer is 0.1-1
mg/dosage unit. In another embodiment, the amount of enhancer is
0.2-1 mg/dosage unit. In another embodiment, the amount is 0.3-1
mg/dosage unit. In another embodiment, the amount is 0.5-1
mg/dosage unit. In another embodiment, the amount is 0.1-2
mg/dosage unit. In another embodiment, the amount is 0.2-2
mg/dosage unit. In another embodiment, the amount is 0.3-2
mg/dosage unit. In another embodiment, the amount is 0.5-2
mg/dosage unit. In another embodiment, the amount is 1-2 mg/dosage
unit. In another embodiment, the amount is 1-10 mg/dosage unit. In
another embodiment, the amount is 2-10 mg/dosage unit. In another
embodiment, the amount is 3-10 mg/dosage unit. In another
embodiment, the amount is 5-10 mg/dosage unit. In another
embodiment, the amount is 1-20 mg/dosage unit. In another
embodiment, the amount is 2-20 mg/dosage unit. In another
embodiment, the amount is 3-20 mg/dosage unit. In another
embodiment, the amount is 5-20 mg/dosage unit. In another
embodiment, the amount is 10-20 mg/dosage unit. In another
embodiment, the amount is 10-100 mg/dosage unit. In another
embodiment, the amount is 20-100 mg/dosage unit. In another
embodiment, the amount is 30-100 mg/dosage unit. In another
embodiment, the amount is 50-100 mg/dosage unit. In another
embodiment, the amount is 10-200 mg/dosage unit. In another
embodiment, the amount is 20-200 mg/dosage unit. In another
embodiment, the amount is 30-200 mg/dosage unit. In another
embodiment, the amount is 50-200 mg/dosage unit. In another
embodiment, the amount is 100-200 mg/dosage unit.
[0055] Each type and amount of enhancer represents a separate
embodiment of the present invention.
[0056] In another embodiment, compositions of the present invention
further comprise a coating that inhibits digestion of the
composition in the stomach of a subject.
[0057] In one embodiment, coating inhibits digestion of the
composition in the stomach of a subject. In one embodiment, the
coated dosage forms of the present invention release drug when pH
move towards alkaline range. In one embodiment, coating is a
monolayer, wherein in other embodiments coating applied in
multilayers. In one embodiment, coating is a bioadhesive polymer
that selectively binds the intestinal mucosa and thus enables drug
release in the attachment site. In one embodiment, the enteric
coating is an enteric film coating. In some embodiment, coating
comprises biodegradable polysaccharide, chitosan, aquateric
aqueous, aquacoat ECD, azo polymer, cellulose acetate phthalate,
cellulose acetate trimelliate, hydroxypropylmethyl cellulose
phthalate, gelatin, poly vinyl acetate phthalate, hydrogel,
pulsincap, or a combination thereof. In one embodiment, pH
sensitive coating will be used according to the desired release
site and/or profile as known to one skilled in the art.
[0058] In one embodiment, the coating is an enteric coating.
Methods for enteric coating are well known in the art, and are
described, for example, in Siepmann F, Siepmann J et al, Blends of
aqueous polymer dispersions used for pellet coating: importance of
the particle size. J Control Release 2005; 105(3): 226-39; and
Huyghebaert N, Vermeire A, Remon J P. In vitro evaluation of
coating polymers for enteric coating and human ileal targeting. Int
J Pharm 2005; 298(1): 26-37. Each method represents a separate
embodiment of the present invention.
[0059] In another embodiment, Eudragit.RTM., an acrylic polymer, is
used as the enteric coating. The use of acrylic polymers for the
coating of pharmaceutical preparations is well known in the art.
Eudragit Acrylic Polymers have been shown to be safe, and are
neither absorbed nor metabolized by the body, but rather are
eliminated.
[0060] In another embodiment, the coating is a gelatin coating. In
another embodiment, microencapsulation is used to protect the
insulin against decomposition in the stomach. Methods for applying
a gelatin coating and for microencapsulation are well known in the
art. Each method represents a separate embodiment of the present
invention.
[0061] In another embodiment, the coating is a film-coating. In
another embodiment, the coating is ethylcellulose. In another
embodiment, the coating is a water-based dispersion of
ethylcellulose, e.g. hydroxypropylmethylcelullose (HPMC) E15. In
another embodiment, the coating is a gastro-resistant coatings,
e.g. a polymer containing carboxylic acid groups as a functional
moiety. In another embodiment, the coating is a monolithic matrix.
In another embodiment, the coating is a cellulose ether (e.g.
hypromellose (HPMC). Each type of coating represents a separate
embodiment of the present invention.
[0062] In another embodiment, a multiparticulate dosage forms is
used to inhibit digestion of the composition in the stomach.
[0063] Each type of coating, dosage form, etc, that inhibits
digestion of the composition in the stomach represents a separate
embodiment of the present invention.
[0064] In another embodiment, the present invention provides a
method for oral administration of a protein with an enzymatic
activity to a subject, whereby a substantial fraction of the
protein retains the enzymatic activity after absorption through an
intestinal mucosal barrier of the subject, comprising administering
orally to the subject a pharmaceutical composition comprising the
protein and an omega-3 fatty acid, thereby orally administering a
protein with an enzymatic activity to a subject.
[0065] In one embodiment, the protein is a recombinant protein. In
one embodiment, the protein is an insulin. In another embodiment,
the protein is a glucagon. In another embodiment, the protein is an
interferon gamma. In another embodiment, the protein is an
interferon alpha. In another embodiment, the protein is a growth
hormone. In another embodiment, the protein is an erythropoietin.
In another embodiment, the protein is granulocyte colony
stimulating factor (G-CSF). In another embodiment, the protein is
any other protein known in the art.
[0066] In another embodiment, the protein is a growth hormone. In
one embodiment, the growth hormone is somatotropin. In another
embodiment, the growth hormone is Insulin Growth Factor-I (IGF-I).
In another embodiment, the growth hormone is any other growth
hormone known in the art.
[0067] In another embodiment, the protein has a molecular weight
(MW) of 1-50 kilodalton (kDa). In another embodiment, the MW is
1-45 kDa. In another embodiment, the MW is 1-40 kDa. In another
embodiment, the MW is 1-35 kDa. In another embodiment, the MW is
1-30 kDa. In another embodiment, the MW is 1-25 kDa. In another
embodiment, the MW is 1-20 kDa. In another embodiment, the MW is
10-50 kDa. In another embodiment, the MW is 15-50 kDa. In another
embodiment, the MW is 20-50 kDa. In another embodiment, the MW is
25-50 kDa. In another embodiment, the MW is 30-50 kDa. In another
embodiment, the MW is 35-50 kDa. In another embodiment, the MW is
1-100 kDa. In another embodiment, the MW is 1-90 kDa. In another
embodiment, the MW is 1-80 kDa. In another embodiment, the MW is
1-70 kDa. In another embodiment, the MW is 1-60 kDa. In another
embodiment, the MW is 10-100 kDa. In another embodiment, the MW is
15-100 kDa. In another embodiment, the MW is 20-100 kDa. In another
embodiment, the MW is 25-100 kDa. In another embodiment, the MW is
30-100 kDa. In another embodiment, the MW is 10-80 kDa. In another
embodiment, the MW is 15-80 kDa. In another embodiment, the MW is
20-80 kDa. In another embodiment, the MW is 25-80 kDa. In another
embodiment, the MW is 30-80 kDa. Each possibility represents a
separate embodiment of the present invention.
[0068] In another embodiment, the MW is less than 20 kDa. In
another embodiment, the MW is less than 25 kDa. In another
embodiment, the MW is less than 30 kDa. In another embodiment, the
MW is less than 35 kDa. In another embodiment, the MW is less than
40 kDa. In another embodiment, the MW is less than 45 kDa. In
another embodiment, the MW is less than 50 kDa. In another
embodiment, the MW is less than 55 kDa. In another embodiment, the
MW is less than 60 kDa. In another embodiment, the MW is less than
65 kDa. In another embodiment, the MW is less than 70 kDa. In
another embodiment, the MW is less than 75 kDa. In another
embodiment, the MW is less than 80 kDa. In another embodiment, the
MW is less than 85 kDa. In another embodiment, the MW is less than
90 kDa. In another embodiment, the MW is less than 95 kDa. In
another embodiment, the MW is less than 100 kDa.
[0069] The molecular weights of some of the proteins mentioned
above are as follows: insulin--6 kilodalton (kDa); glucagon--3.5
kDa; interferon, 28 kDa, growth hormone--21.5-47 kDa; human serum
albumin--69 kDa; erythropoietin--34 kDa; G-CSF-30-34 kDa. Thus, in
one embodiment, the molecular weight of these proteins is
appropriate for administration by methods of the present
invention.
[0070] In another embodiment, methods and compositions of the
present invention are used to administer a human serum albumin.
Human serum albumin is not, in one embodiment, considered to be a
pharmaceutically-active component; however, it can be used in the
context of the present invention as a therapeutically-beneficial
carrier for an active component.
[0071] Each type of protein represents a separate embodiment of the
present invention.
[0072] In another embodiment, the present invention provides a
method for treating diabetes mellitus in a subject, comprising
administering orally to the subject a pharmaceutical composition
comprising an insulin and an omega-3 fatty acid, thereby treating
diabetes mellitus.
[0073] In one embodiment, the diabetes mellitus is Type I diabetes.
In another embodiment, the diabetes mellitus is Type II diabetes.
In another embodiment, the diabetes mellitus is insulin-dependent
diabetes. In another embodiment, the diabetes mellitus is
non-insulin-dependent diabetes. In another embodiment, the diabetes
mellitus is any other type of diabetes known in the art. Each
possibility represents a separate embodiment of the present
invention.
[0074] In one embodiment, three treatments a day of the insulin
composition are administered. In another embodiment, two treatments
a day are administered. In another embodiment, four treatments a
day are administered. In another embodiment, one treatment a day is
administered. In another embodiment, more than four treatments a
day are administered. Each possibility represents a separate
embodiment of the present invention.
[0075] Any of the methods of the present invention may utilize, in
various embodiments, any of the compositions of the present
invention.
[0076] In another embodiment, the present invention provides a
composition for oral administration of insulin, comprising an
insulin protein and an omega-3 fatty acid, whereby a substantial
fraction of the insulin retains the enzymatic activity after
absorption through an intestinal mucosal barrier of the subject
[0077] In one embodiment, the present invention provides a
composition for oral administration of a protein, comprising a
protein and an omega-3 fatty acid, whereby a substantial fraction
of the protein retains the enzymatic activity after absorption
through an intestinal mucosal barrier of the subject.
[0078] In one embodiment, the present invention provides the use of
a protein and an omega-3 fatty acid in the manufacture of a
medicament for oral administration of a protein with an enzymatic
activity to a subject, whereby a substantial fraction of the
protein retains the enzymatic activity after absorption through an
intestinal mucosal barrier of the subject.
[0079] In one embodiment, the present invention provides the use of
an insulin protein and an omega-3 fatty acid in the manufacture of
a medicament for treating diabetes mellitus in a subject.
[0080] In one embodiment, methods and compositions of the present
invention have the advantage of more closely mimicking
physiological insulin secretion by the pancreas. When insulin is
secreted into the portal vein, the liver is exposed to a greater
insulin concentration than peripheral tissues. Similarly, insulin
administered according to the present invention reaches the
intestine and is absorbed in the body through the intestine and
through the portal system to the liver. This absorption route thus
resembles the physiological secretion of insulin by the pancreas,
enabling, in this embodiment, delicate control of the blood glucose
level and the metabolic activities of the liver and the peripheral
organs controlled by insulin. By contrast, when insulin is
administered to insulin-deficient diabetic patients via the
peripheral venous system, the concentration of insulin in the
portal vein is similar to that in the peripheral circulation,
resulting in hypoinsulinemia in the portal vein and the liver and
hyperinsulinemia in the peripheral venous system. This leads, in
one embodiment, to an abnormal pattern of glucose disposal.
[0081] In another embodiment, different constituents of
compositions of the present composition are absorbed at different
rates from the intestinal lumen into the blood stream. The
absorption of the bile acid, in one embodiment, is significantly
faster than the absorption of insulin.
[0082] For this reason, in another embodiment, a drug regimen
involving ingestion of a pair of pills at spaced intervals, e.g., a
second pill containing a higher concentration of enhancer is taken
at a defined interval (e.g. 30 minutes) after the first pill. In
another embodiment, certain of the constituents are
microencapsulated to enhance the absorption of the insulin into the
system.
[0083] In one embodiment, a treatment protocol of the present
invention is therapeutic. In another embodiment, the protocol is
prophylactic. Each possibility represents a separate embodiment of
the present invention.
[0084] In another embodiment, solid carriers/diluents for use in
methods and compositions of the present invention include, but are
not limited to, a gum, a starch (e.g. corn starch, pregeletanized
starch), a sugar (e.g., lactose, mannitol, sucrose, dextrose), a
cellulosic material (e.g. microcrystalline cellulose), an acrylate
(e.g. polymethylacrylate), calcium carbonate, magnesium oxide,
talc, or mixtures thereof.
[0085] In another embodiment, the compositions further comprise
binders (e.g. acacia, cornstarch, gelatin, carbomer, ethyl
cellulose, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl
cellulose, povidone), disintegrating agents (e.g. cornstarch,
potato starch, alginic acid, silicon dioxide, croscarmelose sodium,
crospovidone, guar gum, sodium starch glycolate), buffers (e.g.,
Tris-HCI., acetate, phosphate) of various pH and ionic strength,
additives such as albumin or gelatin to prevent absorption to
surfaces, detergents (e.g., Tween 20, Tween 80, Pluronic F68, bile
acid salts), protease inhibitors, surfactants (e.g. sodium lauryl
sulfate), permeation enhancers, solubilizing agents (e.g.,
glycerol, polyethylene glycerol), anti-oxidants (e.g., ascorbic
acid, sodium metabisulfite, butylated hydroxyanisole), stabilizers
(e.g. hydroxypropyl cellulose, hyroxypropylmethyl cellulose),
viscosity increasing agents (e.g. carbomer, colloidal silicon
dioxide, ethyl cellulose, guar gum), sweeteners (e.g. aspartame,
citric acid), preservatives (e.g., Thimerosal, benzyl alcohol,
parabens), lubricants (e.g. stearic acid, magnesium stearate,
polyethylene glycol, sodium lauryl sulfate), flow-aids (e.g.
colloidal silicon dioxide), plasticizers (e.g. diethyl phthalate,
triethyl citrate), emulsifiers (e.g. carbomer, hydroxypropyl
cellulose, sodium lauryl sulfate), polymer coatings (e.g.,
poloxamers or poloxamines), coating and film forming agents (e.g.
ethyl cellulose, acrylates, polymethacrylates) and/or adjuvants.
Each of the above excipients represents a separate embodiment of
the present invention.
[0086] In some embodiments, the dosage forms of the present
invention are formulated to achieve an immediate release profile,
an extended release profile, or a delayed release profile. In some
embodiments, the release profile of the composition is determined
by using specific excipients that serve for example as binders,
disintegrants, fillers, or coating materials. In one embodiment,
the composition will be formulated to achieve a particular release
profile as known to one skilled in the art.
[0087] In one embodiment, the composition is formulated as an oral
dosage form. In one embodiment, the composition is a solid oral
dosage form comprising tablets, chewable tablets, or capsules. In
one embodiment the capsules are soft gelatin capsules.
[0088] In other embodiments, controlled- or sustained-release
coatings utilized in methods and compositions of the present
invention include formulation in lipophilic depots (e.g. fatty
acids, waxes, oils).
[0089] The compositions also include, in another embodiment,
incorporation of the active material into or onto particulate
preparations of polymeric compounds such as polylactic acid,
polglycolic acid, hydrogels, etc, or onto liposomes,
microemulsions, micelles, unilamellar or multilamellar vesicles,
erythrocyte ghosts, or spheroplasts.) Such compositions will
influence the physical state, solubility, stability, rate of in
vivo release, and rate of in vivo clearance. In another embodiment,
particulate compositions of the active ingredients are coated with
polymers (e.g. poloxamers or poloxamines)
[0090] In another embodiment, the compositions containing the
insulin and omega-3 fatty acid are delivered in a vesicle, e.g. a
liposome (see Langer, Science 249:1527-1533 (1990); Treat et al.,
in Liposomes in the Therapy of Infectious Disease and Cancer,
Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365
(1989); Lopez-Berestein, ibid., pp. 317-327; see generally
ibid).
[0091] The preparation of pharmaceutical compositions that contain
an active component, for example by mixing, granulating, or
tablet-forming processes, is well understood in the art. The active
therapeutic ingredient is often mixed with excipients that are
pharmaceutically acceptable and compatible with the active
ingredient. For oral administration, the active ingredients of
compositions of the present invention are mixed with additives
customary for this purpose, such as vehicles, stabilizers, or inert
diluents, and converted by customary methods into suitable forms
for administration, such as tablets, coated tablets, hard or soft
gelatin capsules, aqueous, alcoholic or oily solutions.
[0092] Each of the above additives, excipients, formulations and
methods of administration represents a separate embodiment of the
present invention.
[0093] In one embodiment, the term "treating" refers to curing a
disease. In another embodiment, "treating" refers to preventing a
disease. In another embodiment, "treating" refers to reducing the
incidence of a disease. In another embodiment, "treating" refers to
ameliorating symptoms of a disease. In another embodiment,
"treating" refers to inducing remission. In another embodiment,
"treating" refers to slowing the progression of a disease.
EXPERIMENTAL DETAILS SECTION
Example 1
Protection of Insulin from Proteases and Successful Administration
Via the Duodenum in Dogs
Materials and Experimental Methods
Formulation
[0094] The day of dosing, a formulation containing 100 milligram
(mg) EDTA (Sigma-Aldrich, St. Louis, Mo.), 100 mg soybean trypsin
inhibitor (SBTI; Sigma), 5 mg insulin (recombinant crystalline)
dissolved in 2 milliliter (ml) fish oil was prepared and inserted
into a transparent gelatin capsule.
Results
[0095] To test whether insulin can be protected from proteases and
absorbed via the duodenum, a composition containing insulin, SBTI,
EDTA, and fish oil was administered directly to the duodenum of an
8.8 kg beagle dog. Blood glucose was measured every 10 minutes
following administration. As depicted below in Table 1, blood
glucose levels were significantly reduced in response to the
insulin.
[0096] Thus, compositions comprising an omega-3 fatty acid can
protect insulin from proteases in the small intestine and enable
direct absorption of orally administered insulin.
TABLE-US-00001 TABLE 1 Blood glucose concentrations following
administration of insulin to the duodenum in experiment #1. Time
(min) Glucose in milligrams/deciliter (mg/dL) -5 67 0 71 10 77 20
62 30 42 40 26 50 41 60 36 75 35 90 51 105 64 120 75
Example 2
Materials and Experimental Methods
Formulation
[0097] 4 days prior to dosing, a formulation was prepared
containing 125 mg EDTA, 100 mg SBTI, and 5 mg insulin in 2 ml fish
oil in a gelatin capsule. The formulation was stored in the
refrigerator (4.degree. C.) until dosing.
Results
[0098] In the next experiment, a formulation of SBTI, EDTA, and
fish oil was prepared 4 days prior to dosing, then administered
directly to the duodenum of a 9.0 kg beagle dog. As depicted below
in Table 2, blood glucose levels were significantly reduced in
response to the insulin.
[0099] These results confirm the results of Example 1, showing that
compositions comprising an omega-3 fatty acid can protect insulin
from proteases in the small intestine and enable direct absorption
of orally administered insulin. In addition, these results show
that compositions of the present invention can be stored after
constitution without losing potency.
TABLE-US-00002 TABLE 2 Blood glucose concentrations following
administration of insulin to the duodenum in experiment #2. Time
(min) Glucose in milligrams/deciliter (mg/dL) -5 69 0 68 10 64 20
38 30 19 40 31 50 39 60 55 75 66 90 75 105 75 120 73
Example 3
Oral Administration of Pills Containing Insulin and Omega-3 Fatty
Acids
Preparation of Tablet Cores
[0100] Tablet cores comprising insulin and omega-3 fatty acids are
prepared using methods well known in the art. For example, tablet
cores may be prepared as described in Example 1.
Coating
[0101] The coating may be any delayed release coating known in the
art. For example, the coating may be a polymer composed of the
following ingredients:
[0102] 4 mg Eudragit L-100 (Polymer of Acrylic and Methacrylic Acid
Esters)
[0103] 4 mg Talc NF
[0104] 0.4 mg Polyethylene Glycol 6000 NF
[0105] In one embodiment, a solution of the enteric coated polymer
is prepared by dissolving the polymer in a methylene
chloride+isopropyl alcohol mixture. The tablets are coated by
spraying the solution within a mildly warmed jar under constant
agitation. The solvent vapors are continuously aspirated.
Measurement of Levels and Activity of Recombinant Insulin in
Subjects' Plasma
[0106] A human insulin radio-immunoassay (RIA) kit (Linco Research
Inc, St. Charles, Mo.) is used to measure levels of recombinant
insulin. Levels of C peptide are measured as well, to determine the
relative contributions of endogenous and exogenous insulin to
observed rises in insulin levels.
Results
[0107] A mixture of EDTA, SBTI, and insulin dissolved in fish oil
is formulated into tablet or capsule cores, coated with an enteric
coating or gelatin coating, and administered to human subjects.
Blood glucose levels of the subjects are measured periodically as
described in the previous Examples. In addition, the subjects'
plasma levels of recombinant insulin and its activity are tested.
The coated pills are shown to deliver functional insulin to the
subjects, and the insulin significantly lowers their blood glucose
levels, showing that active insulin can be delivered to the
bloodstream via oral administration. Different types of
commercially available delayed release coatings are tested to
determine which coating provides the best delivery of insulin, and
this coating is used in subsequent Examples.
Example 4
Optimization of Source of Omega-3 Fatty Acids
[0108] Various omega-3 fatty acids or sources of omega-3 fatty
acids (e.g. those listed above in the specification) are compared
for their ability to preserve insulin following oral administration
in methods and compositions of the present invention. Insulin
tablets or capsules are formulated as described in the above
Examples, except that the insulin is dissolved in the alternate
source instead of in fish oil. The most effective source of omega-3
fatty acids is used in subsequent Examples.
Example 5
Optimization of Protease Inhibitors
[0109] Various protease inhibitors (either non-toxic or having an
acceptable toxicity profile; e.g. those listed above in the
specification) are compared for their ability to preserve insulin
following oral administration in methods and compositions of the
present invention. Insulin tablets or capsules are formulated as
described in the above Examples, except that the alternate protease
inhibitors are substituted for SBTI. Amounts of the protease
inhibitors are also varied, to determine the optimal amounts. The
most effective protease inhibitor/amount is used in subsequent
Examples.
Example 6
Optimization of Enhancer
[0110] Various enhancers (e.g. those listed above in the
specification) are compared for their ability to facilitate
absorption of insulin following oral administration in methods and
compositions of the present invention. Insulin tablets or capsules
are formulated as described in the above Examples, except that the
alternate enhancers are substituted for EDTA. Amounts of the
enhancers are also varied, to determine the optimal amounts. The
most effective enhancer/amount is used in subsequent
experiments.
Example 7
Optimization of Type and Amount of Insulin
[0111] Various types and amounts of insulin e.g. those listed above
in the specification) are compared for their ability to regulate
blood sugar in methods and compositions of the present invention.
Insulin tablets or capsules are formulated as described in the
above Examples, except that the type and amount of insulin is
varied. The most effective type/amount of insulin is used in
clinical trials.
* * * * *