U.S. patent application number 15/333654 was filed with the patent office on 2017-04-06 for compositions and methods relating to ionic salts of peptides.
The applicant listed for this patent is Thetis Pharmaceuticals LLC. Invention is credited to Gary Mathias, Banavara L. Mylari, Frank C. Sciavolino.
Application Number | 20170096449 15/333654 |
Document ID | / |
Family ID | 53267598 |
Filed Date | 2017-04-06 |
United States Patent
Application |
20170096449 |
Kind Code |
A1 |
Mathias; Gary ; et
al. |
April 6, 2017 |
Compositions and Methods Relating to Ionic Salts of Peptides
Abstract
The present invention relates to compounds of Formula I,
compositions containing same, and methods of use.
Inventors: |
Mathias; Gary; (Ridgefield,
CT) ; Mylari; Banavara L.; (Lutz, FL) ;
Sciavolino; Frank C.; (Waterford, CT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Thetis Pharmaceuticals LLC |
Southport |
CT |
US |
|
|
Family ID: |
53267598 |
Appl. No.: |
15/333654 |
Filed: |
October 25, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14772320 |
Sep 2, 2015 |
9505709 |
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PCT/US2015/029091 |
May 4, 2015 |
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15333654 |
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61988721 |
May 5, 2014 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 1/00 20180101; C07C
51/412 20130101; C07C 2601/14 20170501; C07C 53/134 20130101; C07C
317/04 20130101; C07C 51/412 20130101; C07D 213/80 20130101; C07C
57/03 20130101; C07K 5/06095 20130101; A61P 9/00 20180101; A23V
2002/00 20130101; C07C 229/28 20130101; A61P 3/00 20180101; C07C
57/03 20130101; C07C 237/12 20130101; C07K 5/06086 20130101; A23L
33/12 20160801; C07K 7/08 20130101; A61P 25/00 20180101; C11C 3/00
20130101; C07C 57/02 20130101; A61P 29/00 20180101 |
International
Class: |
C07K 5/068 20060101
C07K005/068; A23L 33/12 20060101 A23L033/12; C07D 213/80 20060101
C07D213/80; C07C 53/134 20060101 C07C053/134; C07C 57/03 20060101
C07C057/03; C07C 229/28 20060101 C07C229/28 |
Claims
1.-32. (canceled)
33. A compound of Formula I, or an enantiomer, polymorph, solvate,
or hydrate thereof, which is a salt of an amino acid moiety and one
or two fatty acids, A and B: ##STR00037## wherein A and B are the
same or different, and each of A and B is independently selected
from the group consisting of linoleic acid, gamma-linolenic acid
(GLA), eicosadienoic acid, dihomo-gamma-linolenic acid (DGLA),
arachidonic acid (AA), docosadienoic acid, adrenic acid,
docosapentaenoic acid (Osbond acid), tetracosatetraenoic acid, and
tetracosapentaenoic acid, 24:5 (n-6). either A or B, but not both,
may be absent, X.sub.1 and X.sub.2 are each independently selected
from (CH.sub.2).sub.3--R.sub.1, and (CH.sub.2).sub.4--R.sub.2,
where R.sub.1 and R.sub.2 are each a basic function which may be
the same or different, the basic function being selected from the
group consisting of a primary amine, a secondary amine, a tertiary
amine, and a guanidine, and X.sub.3 is H.
34. The compound of claim 33, wherein X.sub.1 and X.sub.2 are both
(CH.sub.2).sub.4--R.sub.2 and R.sub.2 is NH.sub.3+.
35. The compound of claim 33, wherein X.sub.1 and X.sub.2 are both
(CH.sub.2).sub.3--R.sub.1 and R.sub.1 is
NHC(NH.sub.2+)NH.sub.2.
36. The compound of claim 33, wherein X.sub.1 is
(CH.sub.2).sub.3--R.sub.1, R.sub.1 is NHC(NH.sub.2+)NH.sub.2,
X.sub.2 is (CH.sub.2).sub.4--R.sub.2 and R.sub.2 is NH.sub.3+.
37. The compound of claim 33, wherein X.sub.1 is
(CH.sub.2).sub.4--R.sub.2, R.sub.2 is NH.sub.3+, X.sub.2 is
(CH.sub.2).sub.3--R.sub.1, and R.sub.1 is
NHC(NH.sub.2+)NH.sub.2.
38. A pharmaceutical composition comprising the compound of claim
33, and a carrier.
39. The pharmaceutical composition of claim 38, wherein the
composition is a solid oral dosage form.
40. The pharmaceutical composition of claim 38, wherein the
composition is an intravenous dosage form.
41. The pharmaceutical composition of claim 38, wherein the
composition is an ophthalmic formulation.
42. The pharmaceutical composition of claim 38, wherein the
composition is a parenteral dosage form.
43. A package or kit comprising a unit dosage form of the
pharmaceutical composition of claim 38, at least one container for
holding the unit dosage forms, and instructions for use.
44. The compound of claim 34, wherein A or B is absent and the
remainder is arachidonic acid (AA).
45. The compound of claim 34, wherein A and B are both present and
are each is arachidonic acid (AA).
46. The compound of claim 35, wherein A or B is absent and the
remainder is arachidonic acid (AA).
47. The compound of claim 35, wherein A and B are both present and
are each is arachidonic acid (AA).
48. The compound of claim 36, wherein A or B is absent and the
remainder is arachidonic acid (AA).
49. The compound of claim 36, wherein A and B are both present and
are each is arachidonic acid (AA).
50. The compound of claim 37, wherein A or B is absent and the
remainder is arachidonic acid (AA).
51. The compound of claim 37, wherein A and B are both present and
are each is arachidonic acid (AA).
52. A composition comprising a compound of claim 33.
53. The composition of claim 52, wherein the composition is a
dietary supplement or nutraceutical additive.
54. A method for treating a metabolic disease or disorder in a
human subject, the method comprising administering to the subject a
pharmaceutical composition comprising the compound of claim 33,
wherein the metabolic disease or disorder is selected from the
group consisting of hypertriglyceridemia, severe
hypertriglyceridemia, hypercholesterolemia, pre-diabetes, fatty
liver disease, and obesity.
55. A method for treating a cardiovascular disease or disorder in a
subject, the method comprising administering to the subject a
pharmaceutical composition comprising the compound of claim 33,
wherein the cardiovascular disease or disorder is selected from the
group consisting of atrial fibrillation, myocardial infarction, and
congestive heart failure.
56. A method for treating an inflammatory disease or disorder in a
subject, the method comprising administering to the subject a
pharmaceutical composition comprising the compound of claim 33,
wherein the inflammatory disease or disorder is selected from the
group consisting of arthritis, irritable bowel syndrome, ophthalmic
inflammation disorders, and dry eye syndrome.
57. A method for treating a gastrointestinal disorder or
complication thereof in a subject, the method comprising
administering to the subject a pharmaceutical composition
comprising the compound of claim 33, wherein the gastrointestinal
disorder or complication thereof is selected from the group
consisting of bowel obstruction, short bowel syndrome,
Gastroschisis, prolonged diarrhea, high-output fistula, Crohn's
disease, ulcerative colitis, colon cancer, familial adenomatous
polyposis, parenteral nutrition-associated liver disease, essential
fatty acid deficiency, a congenital gastrointestinal anomaly, and
necrotizing enterocolitis.
58. A method for treating a neurological disorder in a subject, the
method comprising administering to the subject a pharmaceutical
composition comprising the compound of claim 33, wherein the
neurological disorder is selected from the group consisting of
Alzheimer's disease, attention deficit hyperactivity disorder
(ADHD), depression, traumatic brain injury, spinal cord injury,
ischemic stroke, and concussion.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation application of U.S.
patent application Ser. No. 14/772,320, filed on May 4, 2015, which
is the national stage application filed under 35 U.S.C. 371, of
International Application No. PCT/US2015/029091 filed on May 4,
2015, which claims priority to U.S. Provisional Application No.
61/988,721, filed on May 5, 2014. Each of the foregoing disclosures
is incorporated by reference herein in their entireties.
FIELD OF THE INVENTION
[0002] The present invention relates to the field of drug delivery
and particularly the use of peptide-comprising ionic salt compounds
and compositions containing same for delivery of nutraceutical or
therapeutic agents to a subject.
BACKGROUND OF THE INVENTION
[0003] US 20140044828 describes nutritional compositions containing
specified amounts of n-3 fatty acids and one or more of free
lysine, dipeptides containing lysine, and lysine salts, for the
prophylaxis and/or treatment of various symptoms associated with
muscle mass decrease, decreased basal metabolism, and low body
temperature, as well as for suppression of obesity, suppression of
visceral fat accumulation, and treatment of hyperglycemia and
hyperlipidemia, particularly in the elderly. The term "dipeptides
containing lysine" is defined as referring to a dipeptide where at
least one molecule of the dipeptide is lysine or a salt of lysine
such as L-lysine hydrochloride, L-lysine acetate, and L-lysine
glutamate. Lysyllysine is given as a specific example of a
dipeptide containing lysine.
[0004] Lysyllysine is described in US 20080248564 in a process for
the covalent modification of nucleic acids by lactosylation for
more efficient transfer of nucleic acids into cells. This is
described as an improvement over prior art methods relying on the
formation of non-specific ionic complexes between nucleic acids and
polycations such as polylysine, as described in U.S. Pat. No.
5,166,320. US 20060084617 describes the use of lysyllysine in a
process for conjugating endosomolytic spermine to nucleic acids to
enhance their delivery into cells.
[0005] US 20070275019 describes the preparation of vaccines
directed to cancer-associated carbohydrate antigens, the vaccines
comprising multivalent antigen systems in which lysyllysine is used
as a core matrix bearing multiple antigens as dendritic arms
[0006] Polyunsaturated fatty acids of the omega-3 series ("omega-3
fatty acids") have shown a wide spectrum of biological activities
suggesting their possible usefulness in treating a range of
diseases and disorders including metabolic disorders,
cardiovascular complications, inflammatory diseases, central
nervous system disorders, and ophthalmic complications. But the
poor aqueous solubility of omega-3 fatty acids limits their utility
as therapeutic agents and as nutraceutical additives to food and
drink due to a phenomenon referred to as solubility-limited
absorption which limits the plasma levels that can be achieved
following oral administration. In fact, the omega-3 fatty acids are
essentially insoluble in water and both the free acid and sodium
salt forms create soap-like emulsions when mixed with water. Thus,
although omega-3 fatty acids are absorbed following oral
administration, the relatively low plasma levels achieved cannot be
increased simply by increasing the dose administered.
[0007] WO 2014/011895 describes fatty acid salts of
eicosapentaenoic acid (EPA) with lysine or docosahexaenoic acid
(DHA) or EPA with metformin, piperazine, and meglumine. The
compositions provide for increased aqueous solubility of the fatty
acid.
[0008] In addition to their poor aqueous solubility, omega-3 fatty
acids suffer from susceptibility to lipid oxidation. This oxidation
leads to formation of undesirable fishy and rancid off-flavors that
render compositions comprising them less palatable.
[0009] There is a need to develop compositions able to deliver
omega-3 fatty acids at much higher plasma levels than is possible
using the currently available free fatty acid, sodium salt, or
ester forms, in order to fulfill the therapeutic promise of these
compounds and translate the many promising in vitro and cellular
pharmacology observations into clinical benefits. Such compositions
should demonstrate increased aqueous solubility of omeg-3 fatty
acids which would facilitate their use in both oral dosage forms,
ophthalmic drops, and intravenous dosage forms. There is also a
need to develop compositions that provide improved stability of the
omega-3 fatty acids against lipid oxidation. The present invention
addresses these needs.
SUMMARY OF THE INVENTION
[0010] The present invention provides compounds of Formula I or an
enantiomer, polymorph, solvate, or hydrate thereof:
##STR00001## [0011] wherein [0012] A and B are each a molecule
having at least one acidic function, [0013] A and B may be the same
or different, [0014] either A or B, but not both, may be absent,
[0015] X.sub.1 and X.sub.2 each refer to a branched or unbranched
carbon chain of from 1 to 10 carbons comprising at least one basic
function, and [0016] X.sub.3 is H or CO--Z and Z is a peptide of
from 1 to 20 amino acids, or a pharmaceutically acceptable salt
thereof.
[0017] The basic function may be selected from a primary amine, a
secondary amine, a tertiary amine, and a guanidine.
[0018] In one embodiment, X.sub.1 and X.sub.2 are independently
selected from (CH.sub.2).sub.3--R.sub.1, and
(CH.sub.2).sub.4--R.sub.2, where R.sub.1 and R.sub.2 are each a
basic function which may be the same or different. In one
embodiment, X.sub.3 is H. In one embodiment, X.sub.1 and X.sub.2
are both (CH.sub.2).sub.4--R.sub.2, R.sub.2 is NH.sub.3+, and
X.sub.3 is H. In one embodiment, X.sub.1 and X.sub.2 are both
(CH.sub.2).sub.3--R.sub.1, R.sub.1 is NHC(NH.sub.2+)NH.sub.2, and
X.sub.3 is H. In one embodiment, X.sub.1 is
(CH.sub.2).sub.3--R.sub.1, R.sub.1 is NHC(NH.sub.2+)NH.sub.2,
X.sub.2 is (CH.sub.2).sub.4--R.sub.2, R.sub.2 is NH.sub.3+, and
X.sub.3 is H. In one embodiment, X.sub.1 is
(CH.sub.2).sub.4--R.sub.2, R.sub.2 is NH.sub.3+, X.sub.2 is
(CH.sub.2).sub.3--R.sub.1, R.sub.1 is NHC(NH.sub.2+)NH.sub.2, and
X.sub.3 is H.
[0019] In one embodiment, A or B, or both, is a polyunsaturated
fatty acid. In one embodiment, the polyunsaturated fatty acid is an
omega-3 fatty acid selected from the group consisting of
eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA),
docosapentaenoic acid (DPA), hexadecatrienoic acid (HTA),
.alpha.-linolenic acid (ALA), stearidonic acid (SDA),
eicosatrienoic acid (ETE), eicosatetraenoic acid (ETA),
heneicosapentaenoic acid (HPA), tetracosapentaenoic acid, and
tetracosahexaenoic acid. In one embodiment, the polyunsaturated
fatty acid is an omega-6 fatty acid selected from the group
consisting of linoleic acid, gamma-linolenic acid (GLA),
eicosadienoic acid, dihomo-gamma-linolenic acid (DGLA), arachidonic
acid (AA), docosadienoic acid, adrenic acid, docosapentaenoic acid
(Osbond acid), tetracosatetraenoic acid, and tetracosapentaenoic
acid, 24:5 (n-6). In one embodiment, the polyunsaturated fatty acid
is an omega-9 fatty acid selected from the group consisting of mead
acid, 20:3 (n-9), all-cis-5,8,11-eicosatrienoic acid, oleic acid,
eicosenoic acid, erucic acid, and nervonic acid.
[0020] In one embodiment, A or B, or both, are a non-fatty acid
molecule selected from the group consisting of methanesulfonic
acid, niacin, difluoromethylornithine, lipoic acid, gabapentin,
pre-gabalin, indomethacin, sulindac, ibuprofen, naproxen, salicylic
acid, acetylsalicylic acid, salicylsalicylic, and meloxicam.
[0021] The invention also provides compositions comprising a
compound of Formula I. In one embodiment, the composition is a
pharmaceutical or nutriceutical composition and the carrier is
acceptable for administration to humans or animals. In one
embodiment, the composition is a nutriceutical additive or
supplement. In one embodiment, the invention provides a food or
drink product comprising a nutriceutical additive of the invention.
In one embodiment, the composition is a pharmaceutical composition
in the form of a solid oral dosage form, an intravenous dosage
form, or an ophthalmic formulation.
[0022] In one embodiment, the pharmaceutical composition comprises
a compound of Formula I wherein A or B, or both, are a
polyunsaturated fatty acid. In one embodiment, the polyunsaturated
fatty acid is an omega-3 fatty acid independently selected from
eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and
docosapentaenoic acid (DPA). The invention also provides methods of
use for such pharmaceutical compositions. In one embodiment, the
pharmaceutical composition is used in a method for lowering serum
triglycerides in a subject, preferably a human subject. In one
embodiment, the pharmaceutical composition is administered to a
subject in need thereof in an amount effective to lower elevated
serum triglycerides in a human subject by at least 0.5 mmol/L,
preferably at least 1 mmol/L. In one embodiment, the subject is a
human subject having severe hypertriglyceridemia characterized by
serum triglyceride levels of from 500 to 2,000 mg/dl. In one
embodiment, the pharmaceutical composition is used in a method for
treating a metabolic disease or disorder in a subject, preferably a
human subject, the disease or disorder selected from the group
consisting of hypertriglyceridemia, severe hypertriglyceridemia,
hypercholesterolemia, pre-diabetes, fatty liver disease, and
obesity. In one embodiment, the pharmaceutical composition is used
in a method for treating a cardiovascular disease or disorder
selected from atrial fibrillation, myocardial infarction, and
congestive heart failure. In one embodiment, the pharmaceutical
composition is used in a method for treating an inflammatory
disease or disorder selected from arthritis, irritable bowel
syndrome, ophthalmic inflammation disorders, and dry eye syndrome.
In one embodiment, the pharmaceutical composition is used in a
method for treating a gastrointestinal disorder or complication
thereof selected from bowel obstruction, short bowel syndrome,
Gastroschisis, prolonged diarrhea regardless of its cause,
high-output fistula, very severe Crohn's disease, ulcerative
colitis, colon cancer or familial adenomatous polyposis, parenteral
nutrition-associated liver disease, essential fatty acid deficiency
or other pediatric GI disorders including congenital GI anomalies
and necrotizing enterocolitis. In one embodiment, the
pharmaceutical composition is used in a method for treating a
neurological disorder selected from Alzheimer's disease, attention
deficit hyperactivity disorder (ADHD), depression, traumatic brain
injury, spinal cord injury, ischemic stroke, and concussion.
[0023] In one embodiment, the pharmaceutical composition comprises
a compound of Formula I wherein A or B, or both, is selected from
gabapentin and a non-steroidal anti-inflammatory agent (NSAID),
wherein, if only one of A or B is gabapentin or a non-steroidal
anti-inflammatory agent (NSAID), the other is a polyunsaturated
fatty acid, preferably selected from EPA, DHA, or DPA. The
invention also provides methods of use for such pharmaceutical
compositions. In one embodiment, both A and B are gabapentin and
the pharmaceutical composition is used in a method for treating
epilepsy or epileptic syndrome. In one embodiment, one of A or B is
gabapentin and the other is a polyunsaturated fatty acid,
preferably selected from EPA, DHA, or DPA and the pharmaceutical
composition is used in a method for treating nociceptive pain. In
one embodiment, one of A or B is a NSAID and the other is a
polyunsaturated fatty acid, preferably selected from EPA, DHA, or
DPA and the pharmaceutical composition is used in a method for
treating neuropathic pain.
[0024] The invention also provides a package or kit comprising a
unit dosage form of a composition of the invention, at least one
container for holding the unit dosage forms, and instructions for
use.
[0025] The invention also provides a method of achieving an initial
(from time 0.5 to 2 hrs post-administration) free fatty acid plasma
concentration of an omega-3 fatty acid in a subject that is from 2
to 5 times higher than the initial free fatty acid plasma
concentration achievable from administering the free fatty acid
form of the omega-3 fatty acid, the method comprising administering
to the subject by an oral or intravenous route an amount of a
pharmaceutical composition comprising a compound of Formula I
wherein A or B, or both, are a omega-3 fatty acid.
BRIEF DESCRIPTION OF THE FIGURES
[0026] FIG. 1: plasma levels of free EPA of TP-212 compared to
EPA-EE and EPA-FFA, all administered via oral gavage.
[0027] FIG. 2: plasma levels of free DHA of TP-312 compared to DHA
FFA, all administered via oral gavage.
[0028] FIG. 3: plasma levels of free EPA of TP-212 administered via
intravenous injection (TP-212iv) versus TP-212 administered via
oral gavage (TP-212po) versus EPA-FFA administered via oral gavage
(EPApo).
[0029] FIG. 4: plasma levels of free DHA of TP-312 administered via
intravenous injection (TP-312iv) versus TP-312 administered via
oral gavage (TP-312po) versus DHA-FFA administered via oral gavage
(DHApo).
[0030] FIGS. 5A-5C: TP-312-1 Hyperemia (FIG. 5A) Pre-dose, (FIG.
5B) Post-dose, (FIG. 5C) delta.
[0031] FIGS. 6A-6C: TP-312-1 Squinting (FIG. 6A) Pre-dose, (FIG.
6B) Post-dose, (FIG. 6C) delta.
[0032] FIGS. 7A-7C: TP-312-1 Discharge (FIG. 7A) Pre-dose, (FIG.
7B) Post-dose, (FIG. 7C) delta.
[0033] FIGS. 8A-8C: TP-312-1 Lid Swelling (FIG. 8A) Pre-dose, (FIG.
8B) Post-dose, (FIG. 8C) delta.
DETAILED DESCRIPTION OF THE INVENTION
[0034] The invention relates to ionic salt compounds in which the
cation is contributed by a peptide component consisting of at least
two amino acid moieties each having at least one basic function,
and the counter-ion is contributed by a counter-ion component
consisting of one or two molecules, each having at least one acidic
function. Each molecule of the counter-ion component is coordinated
around the at least one basic function of each amino acid moiety of
the peptide component.
[0035] Each amino acid moiety of the peptide component may,
independently, comprise or consist of a single natural or
non-naturally occurring amino acid, or a peptide of from 2 to 5
natural or non-naturally occurring amino acids, or combinations
thereof. In all cases, each amino moiety is either a single amino
acid or a branched or unbranched carbon chain of from 2 to 5
carbons comprising at least one basic function. In one embodiment,
the at least one basic function is selected from the group
consisting of a primary amine, a secondary amine, a tertiary amine,
and a guanidine. In one embodiment, the basic function is a primary
amine. In one embodiment, the primary amine is the terminal amine
of an amino acid side chain, wherein the amino acid is a natural or
non-naturally occurring amino acid. In one embodiment, the amino
acid is a naturally occurring amino acid selected from arginine and
lysine. In one embodiment, the peptide component consists of two
amino acid moieties. In one embodiment, the two amino acid moieties
are independently selected from lysine, arginine, and derivatives
thereof. Preferably, the counterion molecule is a therapeutic
agent. In one aspect, the therapeutic agent is poorly water soluble
and/or chemically unstable, e.g., due to its susceptibility to
oxidative or other degradation. In one embodiment, the counterion
molecule is selected from an omega-3 fatty acid and a non-omega-3
fatty acid therapeutic agent. In one embodiment, the compound has
both an omega-3 fatty acid and a non-omega-3 fatty acid counterion
molecule. The invention also provides compositions comprising same,
and methods of making and using the compositions.
[0036] The present invention provides compounds of Formula I,
including enantiomers, polymorphs, solvates, and hydrates
thereof:
##STR00002## [0037] wherein [0038] A and B are each a molecule
having at least one acidic function, [0039] A and B may be the same
or different, [0040] either A or B, but not both, may be absent,
[0041] X.sub.1 and X.sub.2 each refer to a branched or unbranched
carbon chain of from 1 to 10 carbons comprising at least one basic
function, and [0042] X.sub.3 is H or CO--Z and Z is a peptide of
from 1 to 20 amino acids, or a pharmaceutically acceptable salt
thereof.
[0043] In one embodiment, X.sub.1 and X.sub.2 are independently
selected from (CH.sub.2).sub.3--R.sub.1, and
(CH.sub.2).sub.4--R.sub.2, where R.sub.1 and R.sub.2 are each a
basic function which may be the same or different. In one
embodiment, the basic function is selected from a primary amine, a
secondary amine, a tertiary amine, and a guanidine. In one
embodiment, R.sub.1 is NHC(NH.sub.2+)NH.sub.2. In one embodiment
R.sub.2 is NH.sub.3+.
[0044] In one embodiment, X.sub.1 and X.sub.2 are the same and are
each (CH.sub.2).sub.3--R.sub.1 and R.sub.1 is
NHC(NH.sub.2+)NH.sub.2. In one embodiment, X.sub.1 and X.sub.2 are
the same and are each (CH.sub.2).sub.4--R.sub.2 and R.sub.2 is
NH.sub.3+.
[0045] In one embodiment, X.sub.1 and X.sub.2 are different. In one
embodiment, X.sub.1 is (CH.sub.2).sub.3--R.sub.1, R.sub.1 is
NHC(NH.sub.2+)NH.sub.2, X.sub.2 is (CH.sub.2).sub.4--R.sub.2 and
R.sub.2 is NH.sub.3+. In one embodiment, X.sub.1 is
(CH.sub.2).sub.4--R.sub.2, R.sub.2 is NH.sub.3+, X.sub.2 is
(CH.sub.2).sub.3--R.sub.1, and R.sub.1 is
NHC(NH.sub.2+)NH.sub.2.
[0046] In one embodiment, X.sub.3 is H. In one embodiment, X.sub.3
is H, X.sub.1 is (CH.sub.2).sub.3--R.sub.1, and X.sub.2 is
(CH.sub.2).sub.4--R.sub.2, where R.sub.1 and R.sub.2 are each
NHC(NH.sub.2+)NH.sub.2 and NH.sub.3+, respectively. In one
embodiment, X.sub.3 is H, X.sub.1 is (CH.sub.2).sub.4--R.sub.1,
X.sub.2 is (CH.sub.2).sub.3--R.sub.2, where R.sub.1 and R.sub.2 are
each NH.sub.3+ and NHC(NH.sub.2+)NH.sub.2, respectively. In one
embodiment, X.sub.3 is H, X.sub.1 and X.sub.2 are the same and are
each (CH.sub.2).sub.4--R.sub.2 and R.sub.2 is NH.sub.3+. In one
embodiment, X.sub.3 is H, X.sub.1 and X.sub.2 are the same and are
each (CH.sub.2).sub.3--R.sub.1 and R.sub.1 is
NHC(NH.sub.2+)NH.sub.2.
[0047] In one embodiment, X.sub.3 is CO--Z and Z is a peptide of
from 1 to 20 amino acids. In one embodiment, the peptide is a
peptide of from 1 to 10 or from 1 to 5 amino acids. The amino acids
may be any natural or non-naturally occurring amino acids. In one
embodiment, the amino acids are independently selected from
glycine, alanine, valine, leucine, isoleucine, serine, cysteine,
threonine, methionine, proline, phenylalanine, tyrosine,
tryptophan, histidine, lysine, arginine, aspartic acid, glutamic
acid, asparagine, and glutamine, or salts thereof. The amino salts
may be, for example, the hydrochloride, citrate, tartarate,
monohydrogen-, dihydrogen-, and trihydrogen phosphate,
methanesufonate, benzenesulfonate and borate salt.
[0048] In one embodiment, either A or B is absent. Where either A
or B is absent, the compound may be referred to as "mono" salt. In
one embodiment, A and B are both present. Where A and B are both
present, the compound may be referred to as a "bis" salt.
[0049] In one embodiment, A and B are each a fatty acid and A and B
are the same or different. In another embodiment, A or B is a fatty
acid and the other molecule of the counter-ion component is a
non-fatty acid molecule. In one embodiment, both A and B are a
non-fatty acid molecule. In one embodiment, the molecule is
selected from methanesulfonic acid, niacin, difluoromethylornithine
(also referred to as eflornithine), including its optical forms
(e.g., D, L and racemic mixtures), lipoic acid, including its
optical forms (e.g., D, L and racemic mixtures), gabapentin,
pre-gabalin, indomethacin, sulindac, ibuprofen, naproxen, salicylic
acid, acetylsalicylic acid, salicylsalicylic, and meloxicam. In one
embodiment, the molecule is selected from salicylic acid,
acetylsalicylic acid, and salicylsalicylic. In one embodiment, the
non-fatty acid molecule is a therapeutic agent.
[0050] The term "fatty acid" is used to describe a carboxylic acid
with a long aliphatic carbon chain for from about 4 to 28 carbon
atoms, which is either saturated or unsaturated, referring to
whether the carbon chain contains one or more double bonds between
the carbon atoms (unsaturated). In one embodiment, the fatty acid
is an unsaturated fatty acid. In one embodiment, the unsaturated
fatty acid is a mono-, di-, or polyunsaturated fatty acid. In one
embodiment, the fatty acid is a polyunsaturated fatty acid. In one
embodiment, the polyunsaturated fatty acid is a long-chain
polyunsaturated fatty acid having 16 to 24 carbon atoms
(C.sub.16-C.sub.24), or 20 to 22 carbon atoms (C.sub.20-C.sub.22).
In one embodiment, the polyunsaturated fatty acid is a fatty acid
of the omega-3, omega-6, omega-7, or omega-9 series. In one
embodiment, the fatty acid is selected from a mono-, di-, or
polyunsaturated fatty acid of the omega-3, omega-6, omega-7, or
omega-9 series. Examples of fatty acids of the omega-3, 6, 7, and 9
series are provided in Table 1 below. In one embodiment, the fatty
acid is selected from a fatty acid set forth in Table 1.
TABLE-US-00001 TABLE 1 Fatty acids (mono- and di-unsaturated) of
the omega-3, 6, 7, and 9 series. Common name Lipid name Chemical
name Hexadecatrienoic acid (HTA) 16:3 (n-3)
all-cis-7,10,13-hexadecatrienoic acid .alpha.-Linolenic acid (ALA)
18:3 (n-3) all-cis-9,12,15-octadecatrienoic acid Stearidonic acid
(SDA) 18:4 (n-3) all-cis-6,9,12,15-octadecatetraenoic acid
Eicosatrienoic acid (ETE) 20:3 (n-3)
all-cis-11,14,17-eicosatrienoic acid Eicosatetraenoic acid (ETA)
20:4 (n-3) all-cis-8,11,14,17-eicosatetraenoic acid
Eicosapentaenoic acid (EPA) 20:5 (n-3)
all-cis-5,8,11,14,17-eicosapentaenoic acid Heneicosapentaenoic acid
(HPA) 21:5 (n-3) all-cis-6,9,12,15,18-heneicosapentaenoic acid
Docosapentaenoic acid (DPA), 22:5 (n-3)
all-cis-7,10,13,16,19-docosapentaenoic acid Clupanodonic acid
Docosahexaenoic acid (DHA) 22:6 (n-3)
all-cis-4,7,10,13,16,19-docosahexaenoic acid Tetracosapentaenoic
acid 24:5 (n-3) all-cis-9,12,15,18,21-tetracosapentaenoic acid
Tetracosahexaenoic acid (Nisinic acid) 24:6 (n-3)
all-cis-6,9,12,15,18,21-tetracosahexaenoic acid Linoleic acid (LA)
18:2 (n-6) all-cis-9,12-octadecadienoic acid Gamma-linolenic acid
(GLA) 18:3 (n-6) all-cis-6,9,12-octadecatrienoic acid Calendic acid
18:3 (n-6) 8E,10E,12Z-octadecatrienoic acid Eicosadienoic acid 20:2
(n-6) all-cis-11,14-eicosadienoic acid Dihomo-gamma-linolenic acid
(DGLA) 20:3 (n-6) all-cis-8,11,14-eicosatrienoic acid Arachidonic
acid (AA) 20:4 (n-6) all-cis-5,8,11,14-eicosatetraenoic acid
Docosadienoic acid 22:2 (n-6) all-cis-13,16-docosadienoic acid
Adrenic acid 22:4 (n-6) all-cis-7,10,13,16-docosatetraenoic acid
Docosapentaenoic acid 22:5 (n-6)
all-cis-4,7,10,13,16-docosapentaenoic acid Tetracosatetraenoic acid
24:4 (n-6) all-cis-9,12,15,18-tetracosatetraenoic acid
Tetracosapentaenoic acid 24:5 (n-6)
all-cis-6,9,12,15,18-tetracosapentaenoic acid none 12:1 (n-7)
5-Dodecenoic acid none 14:1 (n-7) 7-Tetradecenoic acid Palmitoleic
acid 16:1 (n-7) 9-Hexadecenoic acid Vaccenic acid 18:1 (n-7)
11-Octadecenoic acid Paullinic acid 20:1 (n-7) 13-Eicosenoic acid
none 22:1 (n-7) 15-Docosenoic acid none 24:1 (n-7) 17-Tetracosenoic
acid oleic acid 18:1 (n-9) 9-octadecenoic acid elaidic acid 18:1
(n-9) (E)-octadec-9-enoic acid gondoic acid 20:1 (n-9)
11-eicosenoic acid mead acid 20:3 (n-9) 5,8,11-eicosatrienoic acid
erucic acid 22:1 (n-9) 13-docosenoic acid nervonic acid 24:1 (n-9)
15-tetracosenoic acid Conjugated Linoleic Acids (two conjugated
double bonds) Rumenic acid 18:2 (n-7) 9Z,11E-octadeca-9,11-dienoic
acid 18:2 (n-6) 10E,12Z-octadeca-9,11-dienoic acid Conjugated
Linolenic Acids (three conjugated double bonds) .alpha.-Calendic
acid 18:3 (n-6) 8E,10E,12Z-octadecatrienoic acid .beta.-Calendic
acid 18:3 (n-6) 8E,10E,12E-octadecatrienoic acid Jacaric acid 18:3
(n-6) 8Z,10E,12Z-octadecatrienoic acid .alpha.-Eleostearic acid
18:3 (n-5) 9Z,11E,13E-octadeca-9,11,13-trienoic acid
.beta.-Eleostearic acid 18:3 (n-5)
9E,11E,13E-octadeca-9,11,13-trienoic acid Catalpic acid 18:3 (n-5)
9Z,11Z,13E-octadeca-9,11,13-trienoic acid Punicic acid 18:3 (n-5)
9Z,11E,13Z-octadeca-9,11,13-trienoic acid Other Rumelenic acid 18:3
(n-3) 9E,11Z,15E-octadeca-9,11,15-trienoic acid .alpha.-Parinaric
acid 18:4 (n-3) 9E,11Z,13Z,15E-octadeca-9,11,13,15-tetraenoic acid
.beta.-Parinaric acid 18:4 (n-3) all
trans-octadeca-9,11,13,15-tretraenoic acid Bosseopentaenoic acid
20:5 (n-6) 5Z,8Z,10E,12E,14Z-eicosanoic acid Pinolenic acid 18:3
(n-6) (5Z,9Z,12Z)-octadeca-5,9,12-trienoic acid Podocarpic acid
20:3 (n-6) (5Z,11Z,14Z)-eicosa-5,11,14-trienoic acid
[0051] In one embodiment, the omega-3 fatty acid is selected from
the group consisting of hexadecatrienoic acid (HTA),
alpha-linolenic acid (ALA), stearidonic acid (SDA), eicosatrienoic
acid (ETE), eicosatetraenoic acid (ETA), eicosapentaenoic acid
(EPA, timnodonic acid), heneicosapentaenoic acid (HPA),
docosapentaenoic acid (DPA, clupanodonic acid), docosahexaenoic
acid (DHA, Cervonic acid), tetracosapentaenoic acid, 24:5 (n-3),
and tetracosahexaenoic acid (Nisinic acid), 24:6 (n-3).
[0052] In one embodiment, the omega-3 fatty acid is selected from
eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and
docosapentaenoic acid (DPA). In one embodiment, the omega-3 fatty
acids are independently selected from EPA, DHA, DPA,
hexadecatrienoic acid (HTA), .alpha.-linolenic acid (ALA),
stearidonic acid (SDA), eicosatrienoic acid (ETE), eicosatetraenoic
acid (ETA), heneicosapentaenoic acid (HPA), tetracosapentaenoic
acid, and tetracosahexaenoic acid. In one embodiment, the
counter-ion component comprises two omega-3 fatty acids that are
the same.
[0053] In accordance with any of the embodiments described herein,
the omega-6 fatty acids may be selected from the group consisting
of linoleic acid, gamma-linolenic acid (GLA), eicosadienoic acid,
dihomo-gamma-linolenic acid (DGLA), arachidonic acid (AA),
docosadienoic acid, adrenic acid, docosapentaenoic acid (Osbond
acid), tetracosatetraenoic acid, and tetracosapentaenoic acid, 24:5
(n-6).
[0054] In accordance with any of the embodiments described herein,
the omega-9 fatty acids may be selected from the group consisting
of mead acid, 20:3 (n-9), all-cis-5,8,11-eicosatrienoic acid, oleic
acid, eicosenoic acid, erucic acid, and nervonic acid.
[0055] In one embodiment, the omega-3 fatty acid component of the
ionic salt is independently selected from eicosapentaenoic acid
(EPA), docosahexaenoic acid (DHA), and docosapentaenoic acid
(DPA).
[0056] In one embodiment, the invention provides a compound of
Formula I selected from the group consisting of Lysyl-lysine EPA,
Lysyl-lysine bis-EPA, Lysyl-lysine DHA, Lysyl-lysine bis DHA,
Lysyl-lysine EPA DHA, Lysyl-lysine bis gabapentin, Lysyl-lysine EPA
niacin, lysyl-lysine EPA ibuprofen, lysyl-lysine EPA
methanesulfonic acid salt, arginyl-lysine bis gabapentin, and
arginyl-arginine bis gabapentin.
[0057] In certain embodiments, the invention provides a solvate of
a compound of Formula I described herein. A "solvate" refers to a
form of salt bound by a non-covalent bond to another molecule (such
as a polar solvent). Such solvates are typically crystalline solids
having a substantially fixed molar ratio of solute and solvent.
When the solvent is water, the solvate formed is a hydrate. Example
hydrates include hemihydrates, mono hydrates, dihydrates, etc.
[0058] In one embodiment, the invention provides a crystalline form
of a compound of Formula I described herein. In one embodiment, the
invention provides a polymorph of an ionic salt described
herein.
Physical Properties
[0059] The compounds of Formula I and the compositions of the
invention possess superior chemical and physical stability, for
example as compared to mixtures of free fatty acids or the ethyl
ester or glyceryl ester forms of the fatty acids. Physically, the
compounds and compositions of the invention are solid, free flowing
substances suitable for formulation into solid dosage forms such as
powders, tablets, capsules or caplets. In addition, the compounds
and compositions of the invention can be readily combined, e.g., by
physical admixture, with other biologically active agents in a
solid dosage form. As such, the compounds and compositions
described here are different from, and advantageous over, other
fatty acid compositions known in the art, which are generally in
the physical form of an oily liquid.
[0060] Thus, the compounds and compositions of the invention
provide a physically and chemically stable form of fatty acids
useful, for example, in the formulation of solid dosage forms of
fatty acids for human and animal consumption. The fatty acid
component of the compounds and compositions of the invention
possesses superior chemical and physical stability compared to the
free fatty acid or ester forms of the fatty acids, e.g., the ethyl
ester, or glyceryl ester forms of the fatty acids. This stability
stems, in part, from the solid state, free-flowing nature of the
compounds and compositions described herein, which are chemically
and physically more stable than the liquid oil form of the free
fatty acids and esters. For example, the fatty acid component of
the solid compounds and compositions described here is relatively
stable against chemical degradation, such as oxidative degradation,
to which the fatty acid oils are highly susceptible. In particular,
the compositions described here are advantageously stable to air,
oxygen, and humidity such that no change in physical properties,
such as flow characteristics, or in chemical properties, as
measured by NMR spectroscopy, occur following days of storage in an
open vial at room temperature and standard humidity.
[0061] The compositions of the invention advantageously provide
unexpectedly high bioavailability of the fatty acid component due
to the tendency of the counter-ion component to completely
dissociate in aqueous media within the pH range of 8.0 to below
1.0, and well within the pH range commonly observed in the stomach
and upper GI tract of most humans and non-human animal species.
[0062] In one embodiment, the compositions of the invention are
able to deliver at least twice as much free fatty acid in serum
during the initial 2 hours following oral or intravenous
administration. Accordingly, the invention also provides methods
for achieving increased bioavailability of free omega-3 fatty acids
following oral or intravenous administration of a composition of
the invention.
[0063] The compounds and compositions of the invention also provide
improved bioavailability of the fatty acid component as compared
to, for example, free fatty acids and esters of the fatty acids.
The pharmacokinetic properties of the compounds and compositions of
the invention relate, in part, to their advantageous property of
completely dissociating into their component ionic species upon
immersion in aqueous media from alkaline to acidic pH, including
gastric or gastrointestinal fluid. Thus, when a composition (or
compound) of the invention is immersed in aqueous media, the
counter-ion component (i.e., the fatty acid) of Formula I
dissociates into its ionic form, i.e., ionic forms of the free
fatty acids. The compounds and compositions of the invention are
therefore useful for delivering fatty acids, and in particular free
fatty acids in their ionic form, to a human or animal subject.
[0064] The solid, free-flowing character of the compounds of the
invention also provides for ease of their formulation in physical
admixture with other biologically active agents in a solid dosage
form. In one embodiment, the solid dosage form is adapted for oral
delivery. The solid dosage form may also be adapted for other
routes of administration, as described infra.
[0065] The compounds of the invention may provide for increased
water solubility and/or stability of a molecule of the counter-ion
component compared to the molecule itself. In one embodiment, the
compounds of the invention allow for the systemic delivery of
higher amounts of a poorly water soluble molecule in the
counter-ion component, when administered to a subject, for example
by an oral or intravenous route, as compared to the molecule
itself. In some embodiments, molecule of the counter-ion component
also has increased bioavailability when administered by an oral or
intravenous route, as compared to the molecule itself.
[0066] In one embodiment, a compound of Formula I having one or two
polyunsaturated fatty acid molecules as the counter-ion component
provides for relatively high aqueous solubility of the fatty acid
molecule compared to, for example, the aqueous solubility of the
free fatty acid or ester form of the fatty acid, including e.g.,
ethyl esters and glycerol esters, such as triglycerol ester,
phosphatidly choline, etc. In one embodiment, the compound is
soluble in water in a range of from about 10 to 100 mg/ml. A
compound having a single fatty acid component A or B (also referred
to herein as a "mono" salt, as opposed to a compound having both A
and B fatty acid components, referred to herein as a "bis" salt)
may have an aqueous solubility in the range of, for example, 40 to
80 mg/ml. Generally, the his salt will have a lower aqueous
solubility than the corresponding mono salt, but in either case the
aqueous solubility is relatively higher than that of the free fatty
acid or ester form of the fatty acid. In one embodiment, a compound
of the invention has an aqueous solubility that is about 2-fold,
about 5-fold, or about 10-fold higher, or more, than that of the
free fatty acid or ester form of the fatty acid.
[0067] In one embodiment, a compound of the invention provides a
solubility of the fatty acid component in aqueous solution that is
from about 50 to 100 times greater than the solubility of the ethyl
ester form of the fatty acid.
[0068] The compounds of the invention having a fatty acid component
may also demonstrate improved physical stability compared to, for
example, the ethyl ester form of the fatty acid. In certain
embodiments, the compounds of the invention also provide for
increased stability against oxidative degradation of the fatty acid
component.
Pharmacokinetic Properties
[0069] The compounds of the invention demonstrate highly favorable
pharmacokinetic properties. For example, the compounds having a
fatty acid molecule in the counter-ion component provide high
levels of serum free fatty acids following oral or intravenous
administration, as discussed in more detail in the examples, infra.
The present invention provides what is believed to be the first
instance of a compound that is suitable for administrating fatty
acids by the intravenous route. The poor water solubility of fatty
acids in general has contraindicated their use in intravenous
formulations. In addition, the compounds of the invention
formulated as oral dosage forms deliver much higher amounts of the
free fatty acid component to the serum than is achievable with oral
administration of, for example, the free fatty acid itself or the
ethyl ester form of the fatty acid. These properties are further
discussed and exemplified in the examples, infra.
Compositions
[0070] The compounds of the invention may be formulated into
pharmaceutical compositions for human or animal use. In one
embodiment, the compounds are formulated into a pharmaceutical
composition comprising a pharmaceutically acceptable carrier, and
optionally one or more pharmaceutically acceptable excipients.
[0071] The term "excipient" refers to an additive that serves some
purpose in the composition other than a carrier, for example as a
stabilizer, taste masking agent (e.g., a sweetener), solubilizing
agent, or suspending agent. Often, a carrier will serve a dual
purpose as a simple carrier or diluent and an excipient. Examples
of pharmaceutically acceptable excipients may thus include
carriers. Non-limiting examples of excipients for use in the
compositions of the invention include sterile liquids, water,
buffered saline, ethanol, polyols (for example, glycerol, propylene
glycol, liquid polyethylene glycol and the like), oils, detergents,
suspending agents, carbohydrates (e.g., glucose, lactose, sucrose
or dextran), antioxidants (e.g., ascorbic acid or glutathione),
chelating agents, low molecular weight proteins, and suitable
mixtures thereof.
[0072] A suitable excipient or carrier is typically a
pharmaceutically acceptable carrier or excipient for use in animals
or humans (or both). The term "pharmaceutically acceptable"
indicates approval by a regulatory agency of the Federal or a state
government or listed in the U.S. Pharmacopeia or other generally
recognized pharmacopeia such as the European Pharmacopeia, for use
in animals, and more particularly in humans. In the context of the
pharmaceutical compositions of the invention, a "carrier" refers
to, for example, a solvent, a diluent, or vehicle with which the
ionic salt of the invention is formulated for delivery.
[0073] Examples of pharmaceutically acceptable carriers for use in
the compositions of the invention include, without limitation,
sterile aqueous and non-aqueous liquids, water, buffered saline,
ethanol, polyols (for example, glycerol, propylene glycol, liquid
polyethylene glycol and the like), and oils, for liquid dosage
forms; or carbohydrates (e.g., glucose, lactose, sucrose or
dextran) for solid dosage forms.
[0074] Further examples of materials which can serve as
pharmaceutically acceptable carriers include: sugars, such as
lactose, glucose and sucrose; starches, such as corn starch and
potato starch; cellulose, and its derivatives, such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa
butter and suppository waxes; oils, such as peanut oil, cottonseed
oil, safflower oil, sesame oil, olive oil, corn oil and soybean
oil; glycols, such as propylene glycol; polyols, such as glycerin,
sorbitol, mannitol and polyethylene glycol; esters, such as ethyl
oleate and ethyllaurate; agar; buffering agents, such as magnesium
hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;
isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer
solutions; and other non-toxic compatible substances employed in
pharmaceutical formulations.
[0075] The compositions of the invention may be formulated in any
suitable form and for any suitable intended route of
administration. Typically, the dosage form is at least in part
determined by the intended route of administration. In one
embodiment, the dosage form is an oral dosage form. The oral dosage
form may be in the form of a solid, such as a tablet, a capsule
containing particulates, liquids, or powders, a lozenge (including
liquid-filled), a gum, or a gel, or in the form of a liquid. In one
embodiment, the dosage form is a solid oral dosage form.
[0076] In another embodiment, the pharmaceutical composition of the
invention is formulated as a gel or cream suitable for topical
administration.
[0077] A composition of the invention may be in the form of a unit
dose. The unit dose may be, for example, in the form of a tablet or
capsule.
Intravenous Formulations
[0078] In one embodiment, the pharmaceutical composition of the
invention is formulated as an intravenous dosage form. In one
embodiment, the intravenous dosage form is in the form of a clear
solution. In one embodiment the composition is in the form of a
lyophilized solid in an ampule suitable for reconstitution with
sterile water-for-injection or aqueous buffer for intravenous
administration. In one embodiment, the composition is in the form
of a nutritional formula for administration of total parenteral
nutrition.
Ophthalmic Formulations
[0079] In one embodiment, A or B, or both, are useful for treating
or ameliorating one or more symptoms of an ocular disease or
disorder, as described in more detail below. Accordingly, the
invention provides compounds of Formula I in a pharmaceutical
composition of the invention suitable for topical administration to
the eye, also referred to as an ophthalmic formulation. The
formulation may be a solution, suspension, or gel suitable for
ocular administration.
[0080] In one embodiment, the ophthalmic formulation is an aqueous
formulation. In one embodiment, the ophthalmic formulation
comprises one or more of glycerin, hypromellose, propylene glycol
or polyethylene glycol. In one embodiment, the ophthalmic
formulation further comprises one or more of polysorbate 80,
carbomer copolymer type A, purified water, sodium hydroxide,
ascorbic acid, benzalkonium chloride, boric acid, dextrose,
disodium phosphate, glycine, magnesium chloride, potassium
chloride, sodium borate, sodium chloride, sodium citrate, sodium
lactate, edetate disodium, hydrochloric acid, sodium hydroxide,
aminornethylpropanol, hydroxypropyl guar, polyquaternium-I, or
sorbitol.
[0081] In one embodiment, the ophthalmic formulation comprises one
or more of surfactants, tonicity agents, buffers, preservatives,
co-solvents and viscosity building agents. Various tonicity agents
may be employed to adjust the tonicity of the composition,
preferably to that of natural tears for ophthalmic compositions.
For example, sodium chloride, potassium chloride, magnesium
chloride, calcium chloride, dextrose and/or mannitol may be added
to the composition to approximate physiological tonicity.
Preferably, the tonicity agent is present in an amount sufficient
to cause the final composition to have an ophthalmically acceptable
osmolality (generally about 150-450 mOsm, preferably 250-350 mOsm).
An appropriate buffer system (e.g., sodium phosphate, sodium
acetate, sodium citrate, sodium borate or boric acid) may be added
to the compositions to prevent pH drift under storage conditions.
The particular concentration will vary, depending on the agent
employed. Preferably, however, the buffer will be chosen to
maintain a target pH within the range of pH 6-7.5.
[0082] Compositions formulated for the treatment of dry eye-type
diseases and disorders may also comprise aqueous carriers designed
to provide immediate, short-term relief of dry eye-type conditions.
Such carriers can be formulated as a phospholipid carrier or an
artificial tears carrier, or mixtures of both. As used herein,
"phospholipid carrier" and "artificial tears carrier" refer to
aqueous compositions which: (i) comprise one or more phospholipids
(in the case of phospholipid carriers) or other compounds, which
lubricate, "wet," approximate the consistency of endogenous tears,
aid in natural tear build-up, or otherwise provide temporary relief
of dry eye symptoms and conditions upon ocular administration; (ii)
are safe; and (iii) provide the appropriate delivery vehicle for
the topical administration of an effective amount of one or more of
the fatty acid salts of the invention.
[0083] Examples or artificial tears compositions useful as
artificial tears carriers include, but are not limited to,
commercial products, such as Tears Naturale.TM., Tears Naturale
n.TM., Tears Naturale Free.TM., and Bion Tears.TM.. (Alcon
Laboratories, Inc., Fort Worth, Tex.). Examples of phospholipid
carrier formulations include those disclosed in U.S. Pat. No.
4,804,539 (Guo et al.), U.S. Pat. No. 4,883,658 (Holly), U.S. Pat.
No. 4,914,088 (Glonek), U.S. Pat. No. 5,075,104 (Gressel et al.),
U.S. Pat. No. 5,278,151 (Korb et al.), U.S. Pat. No. 5,294,607
(Glonek et al.), U.S. Pat. No. 5,371,108 (Korb et al.), U.S. Pat.
No. 5,578,586 (Gionek et al.); the foregoing patents are
incorporated herein by reference to the extent they disclose
phospholipid compositions useful as phospholipid carriers of the
present invention.
[0084] Other compounds designed to lubricate, "wet," approximate
the consistency of endogenous tears, aid in natural tear build-up,
or otherwise provide temporary relief of dry eye symptoms and
conditions upon ocular administration the eye are known in the art.
Such compounds may enhance the viscosity of the composition, and
include, but are not limited to: monomeric polyols, such as,
glycerol, propylene glycol, ethylene glycol; polymeric polyols,
such as, polyethylene glycol, hydroxypropylmethyl cellulose
("HPMC"), carboxy methylcellulose sodium, hydroxy propylcellulose
("HPC"), dextrans, such as, dextran 70; water soluble proteins,
such as gelatin; and vinyl polymers, such as polyvinyl alcohol,
polyvinylpyrrolidone, povidone and carbomers, such as carbomer
934P, carbomer 941, carbomer 940, carbomer 974P.
[0085] Examples of viscosity enhancing agents include, but are not
limited to polysaccharides, such as hyaluronic acid and its salts,
chondroitin sulfate and its salts, dextrans, various polymers of
the cellulose family; vinyl polymers; and acrylic acid polymers. In
general, the phospholipid carrier or artificial tears will exhibit
a viscosity of 1 to 400 centipoises ("cps"). Topical ophthalmic
products are typically packaged in multidose form. Preservatives
may be required to prevent microbial contamination during use.
Suitable preservatives include benzalkonium chloride,
chlorobutanol, benzododecinium bromide, methyl paraben, propyl
paraben, phenylethyl alcohol, edetate disodium, sorbic acid,
polyquaternium-1, or other agents known to those skilled in the
art. Such preservatives are typically employed at a level of from
0.001 to 1.0% w/v. Unit dose compositions of the present invention
will be sterile, but typically unpreserved. Such compositions,
therefore, generally will not contain preservatives.
[0086] Other wetting agents, emulsifiers and lubricants, such as
sodium lauryl sulfate and magnesium stearate, as well as coloring
agents, release agents, coating agents, and perfumingagents,
preservatives and antioxidants can also be present in the
compositions.
[0087] Examples of pharmaceutically acceptable antioxidants
include: water soluble antioxidants, such as ascorbic acid,
cysteine hydrochloride, sodium bisulfate, sodium metabisulfite,
sodium sulfite and the like; oil-soluble antioxidants, such as
ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), lecithin, propyl gallate, a-tocopherol, and
the like; and metal chelating agents, such as citric acid,
ethylenediamine tetraacetic acid (EDT A), sorbitol, tartaric acid,
phosphoric acid, and the like.
[0088] A contact lens may optionally be used to allow for
extravasation of vasoactive substance over a more prolonged time
period. Vasoactive substances such as Thrombin and Thromboxane A
may further induce increase in tear volume via venular
vasoconstriction and increased perfusion through lacrimal,
accessory lacrimal and surface microvessels; where increased
paracellular endothelial openings that increase capillary
permeability can further enhance this benefit.
[0089] Methods of preparing these formulations or compositions
include the step of bringing into association a compound of the
present invention with the carrier and, optionally, one or more
accessory ingredients. In general, the formulations are prepared by
uniformly and intimately bringing into association a compound of
the present invention with liquid carriers.
Additives and Supplements
[0090] In one embodiment, A or B, or both are useful as a dietary
supplement or nutraceutical additive. For example, fatty acids,
especially polyunsaturated fatty acids of the omega-3, omega-6,
omega-7, and omega-9 series are known to be useful in this context.
Thus, the invention also provides a compound of Formula I
formulated as a nutraceutical additive or supplement, either alone
or in combination with one or more additives or supplements and any
suitable excipients. In one embodiment, the nutraceutical additive
or supplement is in the form of a powder. In one embodiment, the
nutraceutical additive or supplement is in the form of a liquid. In
one embodiment, the nutraceutical additive or supplement is in the
form of a mouth wash, a dentifrice, chewing gum, a candy, a tablet,
a capsule, a mouth spray, or a film.
[0091] In one embodiment, the nutraceutical additive forms part of
a food or drink product suitable for human consumption. There is no
specific limitation on the foods/drinks to which a nutraceutical
additive of the invention can be incorporated. Examples of such
foods/drinks include processed foods based on meat, poultry meat,
fish/shellfish and the like; soup; seasonings including sweetener
and the like; rice seasonings; instant foods; frozen foods; snacks;
various types of functional foods such as supplements, nutritional
drinks and the like; canned foods; dairy products; confectionery
such as chewing gum, candy, gummy candy, chocolate, baked sweets
and the like; ice cream; soft drinks such as tea, coffee, cocoa,
fruit juice, sports drink, carbonated drink, vegetable drink and
the like; liquors; soya milk; lactic acid bacteria beverages; and
chlorophyll juice.
[0092] The amount of the nutraceutical additive of the invention
incorporated into the food or drink varies in accordance with the
type of food or drink and the amount that one wishes to supplement
a diet with one or more omega-3 fatty acids. In one embodiment, the
nutraceutical additive is incorporated into the food or drink so as
to provide an amount of the omega-3 fatty acid that is about
0.000001 to 20% by weight, based on total weight of the food or
drink product, and more preferably in an amount of about 0.00001 to
10% by weight.
Methods of Making
[0093] The compounds of the present invention can be prepared
according to the general Scheme 1 and as further exemplified in
Scheme 2. Further details for the preparation of the compounds are
provided in the Examples section, infra.
##STR00003##
##STR00004##
Methods of Use
[0094] The compositions of the invention are useful in methods of
treating various diseases and disorders that are responsive to
treatment with fatty acids, especially polyunsaturated fatty acids,
and particularly polyunsaturated fatty acids of the omega-3,
omega-6, omega-7, and omega-9 series. In the context of any of the
methods described here, a composition of the invention may be
formulated as a pharmaceutical composition, meaning that the
composition itself and any additives or excipients in the
formulation are suitable for administration to humans or
animals.
[0095] In the context of the methods described here, the term
"treating" may refer to the amelioration or stabilization of one or
more symptoms associated with the disease or disorder. The term
"treating" may also encompass the management of a disease or
disorder, referring to the beneficial effects that a subject
derives from a therapy which does not result in a cure of the
underlying disease or disorder. For example, lowering elevated
plasma triglycerides can be considered an aspect of treating
diabetes because it is a beneficial effect that does not result in
a cure of the underlying defect of glucose metabolism. The
compositions of the invention can also be used in the prevention of
certain diseases, disorders, and conditions. In this context, the
term "prevention" refers to preventing the recurrence, development,
progression or onset of one or more symptoms of the disease,
disorder, or condition.
[0096] In accordance with the methods of the invention, a
therapeutically effective amount of a composition of the invention
is administered to a subject, preferably a human subject, the
therapeutically effective amount being the amount sufficient to
achieve a desired therapeutic outcome, for example the amelioration
or stabilization of one or more symptoms of the disease or disorder
being treated, or in the context of prevention, the amount
sufficient to achieve prevention of the recurrence, development,
progression or onset of one or more symptoms of the disease,
disorder, or condition.
[0097] For administration to human patients, the total daily dose
of the compounds of the invention is typically in the range 1 g to
12 g depending, of course, on the mode of administration. In one
embodiment the total daily dose is in the range 1 g to 10 g, in
another embodiment the total daily dose is in the range 4 g to 8 g
and in yet another embodiment the total daily dose is in the range
1 g to 2 g. The total daily dose may be administered in single or
divided doses.
[0098] These dosages are based on an average human subject having a
weight of about 65 kg to 70 kg. The physician will readily be able
to determine doses for subjects whose weight falls outside this
range, such as infants and the elderly.
[0099] In one embodiment, a therapeutically effective amount is the
amount required to achieve at least an equivalent therapeutic
effect compared to a standard therapy. An example of a standard
therapy is an FDA-approved drug indicated for treating a particular
disease or disorder. As an example, Vascepa.TM. is an FDA-approved
formulation of EPA, specifically an ethyl ester of EPA. Thus, in
one embodiment, the invention provides a pharmaceutical composition
comprising a compound of Formula I having one or two EPA molecules
as the counter-ion component in a therapeutically effective amount,
which amount is effective to reduce plasma triglycerides in an
adult human subject by at least about 0.5 mmol/L, about 1 mmol/L,
or about 2 mmol/L.
[0100] In the context of any of the methods of the present
invention, the subject may be a human or a non-human mammal. The
non-human mammal may be, for example, a non-human primate, a dog,
cat, a rodent (e.g., a mouse, a rat, a rabbit), a horse, a cow, a
sheep, a goat, a bird, a chicken, or any other non-human mammal
Preferably, the subject is a human.
[0101] In one embodiment, the subject is a human subject. In one
embodiment, the human is an adult human, a pediatric human, or a
geriatric human, as those terms are understood by the medical
practitioner, for example as defined by the U.S. Food and Drug
Administration.
[0102] The compositions of the invention can be used as monotherapy
or adjunctive therapy. The compositions of the invention can be
administered alone or in combination with one or more additional
therapeutic agents (i.e., additional APIs) or therapies, for
example as part of a therapeutic regimen that includes, e.g.,
aspects of diet and exercise. In certain embodiments, the methods
of the invention include administration of a composition of the
invention as the primary therapy. In other embodiments, the
administration of a composition of the invention is an adjuvant
therapy. In either case, the methods of the invention contemplate
the administration of a composition of the invention in combination
with one or more additional therapeutic agents and/or therapies for
the treatment or prevention of a disease or disorder. The terms
"therapy" and "therapies" refer to any method, protocol and/or
agent that can be used in the prevention, treatment, management or
amelioration of a disease or disorder, or one or more symptoms
thereof.
Metabolic Disorders
[0103] In one embodiment, the invention provides methods of
treating a metabolic disorder in a subject in need thereof, the
method comprising administering to the subject, preferably a human
subject, a composition comprising a compound of Formula I having
one or two polyunsaturated fatty acid molecules as the counter-ion
component. In one embodiment, the polyunsaturated fatty acid
molecules are of the omega-3, omega-6, omega-7, or omega-9 series.
In one embodiment, the fatty acid molecules are omega-3 fatty acids
independently selected from eicosapentaenoic acid (EPA),
docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA).
[0104] In one embodiment the metabolic disorder is selected from
the group consisting of abnormal glucose metabolism manifesting in
diabetes or pre-diabetes, abnormal lipid metabolism manifesting as
hypertriglyceridemia, i.e., elevated triglycerides, mixed
dyslipidemia, hypercholesterolemia, fatty liver, and combined
abnormal glucose and lipid metabolism manifesting in obesity. In
one embodiment the metabolic disorder is a dyslipidemic disorder
selected from hypertriglyceridemia, hypercholesterolemia and mixed
dyslipidemias. In one embodiment, the metabolic disorder is
selected from the group consisting of pre-diabetes, type 2
diabetes, obesity, fatty liver disease, and insulin resistance.
[0105] In one embodiment, the methods comprise administering a
therapeutically effective amount, which amount is effective to
reduce plasma triglycerides in an adult human subject by at least
about 0.5 mmol/L, about 1 mmol/L, or about 2 mmol/L.
[0106] In one embodiment, the subject is a human subject having
severe hypertriglyceridemia characterized by serum triglyceride
levels of from 500 to 2,000 mg/dl.
Cardiovascular Disorders
[0107] In one embodiment, the invention provides a method for
treating cardiovascular disorders or complications relating to
atrial fibrillation, myocardial infarction, and congestive heart
failure by administering to a subject in need of such treatment an
effective amount of a composition comprising a compound of Formula
I having one or two polyunsaturated fatty acid molecules as the
counter-ion component. In one embodiment, the polyunsaturated fatty
acid molecules are of the omega-3, omega-6, omega-7, or omega-9
series. In one embodiment, the fatty acid molecules are omega-3
fatty acids independently selected from eicosapentaenoic acid
(EPA), docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA).
In one embodiment, the effective amount is effective to treat one
or more symptoms of the cardiovascular condition.
Gastrointestinal Disorders
[0108] In one embodiment, the invention provides a method for
treating gastrointestinal disorders or complications relating to
parenteral nutrition-associated liver disease, and essential fatty
acid deficiency and other pediatric GI disorders including
congenital GI anomalies and necrotizing enterocolitis by
administering to a subject in need of such treatment an effective
amount of a composition comprising a compound of Formula I having
one or two polyunsaturated fatty acid molecules as the counter-ion
component. In one embodiment, the polyunsaturated fatty acid
molecules are of the omega-3, omega-6, omega-7, or omega-9 series.
In one embodiment, the fatty acid molecules are omega-3 fatty acids
independently selected from eicosapentaenoic acid (EPA),
docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA). In one
embodiment, the effective amount is effective to treat one or more
symptoms of the gastrointestinal condition.
[0109] In one embodiment, the invention provides a method for
providing nutrition to patients who do not have a functioning
gastrointestinal tract or who have disorders requiring complete
bowel rest, including bowel obstruction, short bowel syndrome,
Gastroschisis, prolonged diarrhea regardless of its cause,
high-output fistula, and very severe Crohn's disease or ulcerative
colitis by administering to a subject in need of such treatment an
effective amount of a composition comprising a compound of Formula
I having one or two polyunsaturated fatty acid molecules as the
counter-ion component. In one embodiment, the polyunsaturated fatty
acid molecules are of the omega-3, omega-6, omega-7, or omega-9
series. In one embodiment, the fatty acid molecules are omega-3
fatty acids independently selected from eicosapentaenoic acid
(EPA), docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA).
In one embodiment, the effective amount is effective to treat one
or more symptoms of the gastrointestinal condition.
[0110] In one embodiment, the invention provides a method for
preventing cancer, the method comprising administering a
therapeutically effective amount of a composition of the invention
to a subject in need of preventive anti-cancer therapy. In one
embodiment, the cancer is colon cancer or familial adenomatous
polyposis.
Inflammatory Disorders
[0111] The compounds of Formula I having one or two polyunsaturated
fatty acid molecules as the counter-ion component may be
particularly useful in the treatment of diseases and disorders
having a significant inflammatory component, due to the
anti-inflammatory properties of polyunsaturated fatty acids and the
ability of the compounds of Formula I to deliver high amounts of
free fatty acids to the serum by either oral or intravenous routes
of administration.
[0112] In one embodiment, the invention provides a method for
treating an inflammatory disorder, the method comprising
administering to a subject in need of such treatment an effective
amount of a composition comprising a compound of Formula I having
one or two polyunsaturated fatty acid molecules as the counter-ion
component. In one embodiment, the polyunsaturated fatty acid
molecules are of the omega-3, omega-6, omega-7, or omega-9 series.
In one embodiment, the fatty acid molecules are omega-3 fatty acids
independently selected from eicosapentaenoic acid (EPA),
docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA). In one
embodiment, the effective amount is effective to treat one or more
symptoms of the inflammatory disorder. In one embodiment, the
inflammatory disorder is selected from the group consisting of
arthritis, inflammatory bowel disease, and psoriasis.
[0113] In one embodiment, the invention provides methods of
treating arthritis, irritable bowel syndrome, ophthalmic
inflammation disorders, or dry eye syndrome in a subject in need of
such treatment, the methods comprising administering to the subject
a composition comprising a compound of Formula I having one or two
polyunsaturated fatty acid molecules as the counter-ion component.
In one embodiment, the polyunsaturated fatty acid molecules are of
the omega-3, omega-6, omega-7, or omega-9 series. In one
embodiment, the fatty acid molecules are omega-3 fatty acids
independently selected from eicosapentaenoic acid (EPA),
docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA).
[0114] In one embodiment, the invention provides a method for
treating a disease or disorder of the ocular system, also referred
to as ophthalmic diseases and disorders, having an underlying
inflammatory component, the method comprising administering to a
subject in need of such treatment an effective amount of a
composition comprising a compound of Formula I having one or two
polyunsaturated fatty acid molecules as the counter-ion component.
In one embodiment, the polyunsaturated fatty acid molecules are of
the omega-3, omega-6, omega-7, or omega-9 series. In one
embodiment, the fatty acid molecules are omega-3 fatty acids
independently selected from eicosapentaenoic acid (EPA),
docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA). In one
embodiment, the effective amount is effective to treat one or more
symptoms of the disease or disorder of the ocular system. In one
embodiment, the disease or disorder of the ocular system is
selected from the group consisting of inflammatory diseases of the
eye, dry eye syndrome, macular edema and retinopathy. In one
embodiment, the method is a method for promoting corneal wound
healing.
[0115] In one embodiment, the invention provides a method for
treating dry eye by administering a composition comprising a
compound of Formula I having one or two polyunsaturated fatty acid
molecules as the counter-ion component. In one embodiment, the
polyunsaturated fatty acid molecules are of the omega-3, omega-6,
omega-7, or omega-9 series. In one embodiment, the fatty acid
molecules are omega-3 fatty acids independently selected from
eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and
docosapentaenoic acid (DPA). Dry eye disease or syndrome is a
multifactorial disorder of the tears and ocular
surfacecharacterized by symptoms of dryness and irritation.
Inflammation is an important component in the development and
propagation of dry eye (Stevenson et al., Arch. Ophthalmol., 2012,
130(1), 90-100; Rashid et al., Arch. Ophthalmol, 2008, 126(2),
219-225).
[0116] The term"dry eye" refers to inadequate tear production
and/or abnormal tear composition. Causes of dry eye disease as
defined herein include but are not limited to the following:
idiopathic, congenital alacrima, xerophthalmia, lacrimal gland
ablation, and sensory denervation; collagen vascular diseases,
including rheumatoid arthritis, Wegener's granulomatosis, and
systemic lupus erythematosus; Sjogren's syndrome and autoimmune
diseases associated with Sjogren's syndrome; abnormalities of the
lipid tear layer caused by blepharitis or rosacea; abnormalities of
the mucin tear layer caused by vitamin A deficiency; trachoma,
diphtheric keratoconjunctivitis; mucocutaneous disorders; aging;
menopause; and diabetes. Further, the term "dry eye" includes dry
eye after anterior ophthalmic operation such as cataract operation
and refractive surgery and that accompanied with allergic
conjunctivitis Dry eye symptoms as defined herein may also be
provoked by other circumstances, including, but not limited to, the
following: prolonged visual tasking; working on a computer; being
in a dry environment; ocular irritation; contact lenses, LASIK and
other refractive surgeries; fatigue; and medications such as
isotretinoin, sedatives, diuretics, tricyclic antidepressants,
antihypertensives, oral contraceptives, antihistamines, nasal
decongestants, beta-blockers, phenothiazines, atropine, and pain
relieving opiates such as morphine.
Neurological Disorders
[0117] In one embodiment, the invention provides a method for
treating a psychiatric disorder in a subject, the method comprising
administering the subject a therapeutically effect amount of a
composition comprising a compound of Formula I having one or two
polyunsaturated fatty acid molecules as the counter-ion component.
In one embodiment, the polyunsaturated fatty acid molecules are of
the omega-3, omega-6, omega-7, or omega-9 series. In one
embodiment, the fatty acid molecules are omega-3 fatty acids
independently selected from eicosapentaenoic acid (EPA),
docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA), where
the amount is effective to treat one or more symptoms of the
psychiatric disorder. In one embodiment, the psychiatric disorder
is selected from Alzheimer's disease, attention deficit
hyperactivity disorder (ADHD) and depression.
[0118] In one embodiment, the invention provides a method for
treating a neuro trauma injury in a subject, the method comprising
administering to the subject a therapeutically effective amount of
a composition comprising a compound of Formula I having one or two
polyunsaturated fatty acid molecules as the counter-ion component.
In one embodiment, the polyunsaturated fatty acid molecules are of
the omega-3, omega-6, omega-7, or omega-9 series. In one
embodiment, the fatty acid molecules are omega-3 fatty acids
independently selected from eicosapentaenoic acid (EPA),
docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA), where
the amount is effective to treat one or more symptoms of the neuro
trauma injury. In one embodiment, the neuro trauma injury is
selected from traumatic brain injury, spinal cord injury, ischemic
stroke, and concussion.
[0119] In one embodiment, the invention provides a method for
treating epilepsy or epileptic syndrome by administering to a
subject in need of such treatment a composition comprising a
compound of Formula I having one or two gabapentin molecules as the
counter-ion component. In one embodiment, the method comprises
administering to the subject in need of treatment for epilepsy or
epileptic syndrome a composition of the invention which has been
formulated to contain at least one additional API in a single
dosage form. In one embodiment, the additional API is an
anti-epileptic agent such as gabapentin, or a pharmaceutically
acceptable salt and prodrug thereof.
Pain
[0120] In one embodiment, the invention provides a method for
treating or managing pain. In one embodiment, the pain is
neuropathic pain and the method comprises administering to a
subject in need of treatment for neuropathic pain a pharmaceutical
composition comprising a compound of Formula I wherein A and B are
both present, A or B is a polyunsaturated fatty acid, for example,
EPA, DHA, or DPA, and the other molecule of the counter-ion
component is a non-steroidal anti-inflammatory agent (NSAID), or a
pharmaceutically acceptable salt or prodrug thereof.
[0121] In one embodiment, the pain is nociceptive pain and the
method comprises administering to a subject in need of treatment
for nociceptive pain a pharmaceutical composition comprising a
compound of Formula I wherein A and B are both present, A or B is a
polyunsaturated fatty acid, for example, EPA, DHA, or DPA, and the
other molecule of the counter-ion component is gabapentin, or a
pharmaceutically acceptable salt or prodrug thereof.
[0122] In one embodiment, the method comprises administering to the
subject in need of treatment for neuropathic pain a composition of
the invention which has been formulated to contain at least one
additional active pharmaceutical agent (API) in a single dosage
form. In one embodiment, the additional API is a NSAID, or a
pharmaceutically acceptable salt or prodrug thereof. In another
embodiment, the pain is nociceptive pain and the method comprises
administering a composition of the invention to a subject in need
of treatment for nociceptive pain. In one embodiment, the method
comprises administering to the subject in need of treatment for
nociceptive pain a composition of the invention which has been
formulated to contain at least one additional API in a single
dosage form. In one embodiment, the additional API is gabapentin,
or a pharmaceutically acceptable salt or prodrug thereof.
Combination Therapies
[0123] In the context of combination therapies, a composition of
the invention may be administered together with at least one
additional API or separately from the additional API. Where
delivery is together, a composition of the invention may be
delivered in the same dosage form as the additional API, or in a
different dosage form. One of the advantages of the present
invention, as discussed above, is the ease of formulating the
compositions described herein with additional APIs and excipients
in a single solid dosage form due to their form as a free flowing
powder that is chemically and physically stable (as opposed to the
relatively unstable oily liquid form of free fatty acids and their
esters).
[0124] In one embodiment, a composition of the invention is
formulated in a single solid dosage form with an antihyperlipidemic
agent or an anti-diabetic agent. Antihyperlipidemic agents that may
be used include HMG CoA enzyme inhibitors (e.g., statins),
cholesterol absorption inhibitors, and cholesterol esterase
transfer protein (CETP) inhibitors. In one embodiment, the
antihyperlipidemic agent is selected from a statin, a cholesterol
absorption inhibitor, a CETP inhibitor, and
pharmaceutically-acceptable salts and prodrugs of any of the
foregoing. The pharmaceutically acceptable salt may be selected
from the group consisting of a propionate, decanoate, caprylate,
acrylate, formate, isobutyrate, caprate, heptanoate, propiolate,
oxalate, malonate, succinate, suberate, sebacate, fumarate,
maleate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate,
chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate,
methoxybenzoate, phthalate, terephathal ate, sulfonate,
xylenesulfonate, phenyl acetate, phenylpropionate, phenylbutyrate,
citrate, lactate, p-hydroxybutyrate, glycolate, tartrate,
methanesulfonate, propanesulfonates, naphthalene-1-sulfonate,
naphthalene-2-sulfonate, mandelate, hippurate, gluconate, and
lactobionate salt.
[0125] In one embodiment, the antihyperlipidemic agent is a statin.
In one embodiment, the statin is selected from the group consisting
of atorvastatin, risuvostatin, simvastatin, pravastatin, and
pharmaceutically acceptable salts and prodrugs of any of the
foregoing. In one embodiment, the statin is present in an amount
ranging from 5 mg to 100 mg. In one embodiment, the statin is
pravastatin.
[0126] In one embodiment, the antihyperlipidemic agent is a
cholesterol absorption inhibitor. In one embodiment, the
cholesterol absorption inhibitor is ezetimibe, also known as
Zetia.
[0127] In one embodiment, the antihyperlipidemic agent is a CETP
inhibitor. In one embodiment, the CETP inhibitor is anacetrapib, or
a hydrate, or solvate thereof.
[0128] In one embodiment, a composition of the invention is
formulated in a single solid dosage form with an anti-epileptic
agent or an inhibitor of neuropathic pain such as gabapentin, or a
pharmaceutically acceptable salt and prodrug thereof.
[0129] The invention is further described in the following
examples, which do not limit the scope of the invention described
in the claims.
EXAMPLES
[0130] Table 2 shows a number of specific examples of the compounds
described herein.
TABLE-US-00002 TABLE 2 Compounds Com- pound # Structure Name 1
##STR00005## ##STR00006## (L,L) Lysyl-lysine EPA salt (mono) 2
##STR00007## ##STR00008## (L,L) Lysyl-lysine bis-EPA 3 ##STR00009##
##STR00010## (L,L) Lysyl-lysine DHA (mono) 4 ##STR00011##
##STR00012## (L,L) Lysyl-lysine bis DHA 5 ##STR00013## ##STR00014##
(L,L) Lysyl-lysine EPA DHA 6 ##STR00015## ##STR00016## (L,L)
Lysyl-lysine bis gabapentin 7 ##STR00017## ##STR00018## (L,L)
Lysyl-lysine EPA niacin 8 ##STR00019## ##STR00020##
(L,L)-lysyl-lysine EPA ibuprofen 9 ##STR00021## ##STR00022##
(L,L)-lysyl-lysine EPA methanesulfonic acid salt 10 ##STR00023##
##STR00024## (L,L) arginyl-lysine bis gabapentin 11 ##STR00025##
##STR00026## (L,L) arginyl- arginine bis gabapentin
Experimental Procedures for Making Compounds 1-11
[0131] The following precursors are used in making Compounds
1-11.
a. benzyl
N6-((benzyloxy)carbonyl)-N2-(N2,N6-bis((benzyloxy)carbonyl)-L-ly-
syl)-L-lysinate
##STR00027##
[0133] A solution/suspension of H-Lys(Z)-OBzl hydrochloride (16.3
g, 40 mmol) and Z-Lys(Z)--OH (16.6 g, 40 mmol) in anhydrous
dichloromethane (50 mL) under nitrogen was cooled on ice and
treated with HOBT hydrate (9.2 g, 60 mmol) and triethylamine (16.8
mL, 120 mmol), and the clear solution was stirred for 15 min EDC
hydrochloride (9.6 g, 50 mmol) was added, and the stirred mixture
allowed to warm to room temperature and stirred 20 h. The product
mixture was diluted to 500 mL total volume with dichloromethane,
then washed successively with 5% citric acid, water, saturated
aqueous sodium bicarbonate, and brine (200 mL each), and dried
(Na.sub.2SO.sub.4). The solution was added directly to a silica gel
column (.about.500 cc) and eluted with dichloromethane, then 2:1
dichloromethane/ethyl acetate to afford 29.71 g (97%) of subject
material as a white solid. MP 145-6.degree. C. NMR (CDCl.sub.3):
.delta. 7.15-7.30 (m, 20H), 6.50-6.65 (m, 1H), 5.40-5.55 (m, 1H),
5.10 (d, 1H, J=11.5 Hz), 4.90-5.10 (m, 8H), 4.45-4.55 (m, 1H),
4.05-4.15 (m, 1H), 2.90-3.15 (m, 4H), 1.70-1.80 (m, 2H), 1.55-1.65
(m, 2H), 1.10-1.45 (m, 8H). .sup.13C NMR (151 MHz, cdcl.sub.3)
.delta. 171.94, 156.68, 156.59, 156.33, 136.59, 136.50, 136.15,
135.23, 128.60, 128.59, 128.50, 128.48, 128.47, 128.37, 128.34,
128.16, 128.09, 128.04, 76.82, 67.21, 67.05, 66.69, 66.60, 54.50,
52.14, 40.22, 40.17, 32.06, 31.25, 29.17, 22.14, 22.04.
b. L-lysyl-L-lysine
##STR00028##
[0135] A stirred solution/suspension of (L,L)-Z3-lysyllisine,
benzyl ester (5.37 g, 7.0 mmol) in methanol (120 mL) under nitrogen
was treated with 10% Pd/C (0.50 g), then evacuated and purged
several times with hydrogen via balloon. The mixture was stirred
for 42 h under hydrogen, then the flask evacuated with nitrogen and
carefully filtered through Celite with water rinse. The filtrate
was concentrated in vacuo to afford a white foam. The foam was
transferred to a 50 mL pear shaped flask by dissolving in hot
methanol, then concentrated in vacuo and the residual foam stirred
with acetonitrile for 30 min, filtered, collected, and dried in
vacuo to afford 1.92 g (100%) as a white powder. MP 88-90.degree.
C. NMR (D.sub.2O): .delta. 4.08 (dd, 1H, J=5 Hz, 8 Hz), 3.31 (t,
1H, J=7 Hz), 2.75-2.95 (m, 4H), 1.45-1.80 (m, 8H), 1.20-1.40 (m,
4H). .sup.13C NMR (151 MHz, d.sub.2o) .delta. 178.75, 176.84,
54.77, 54.26, 40.95, 39.40, 33.72, 31.12, 27.66, 27.11, 22.14,
21.80.
c. benzyl
N6-((benzyloxy)carbonyl)-N2-(N2,Nd,Nw-tris((benzyloxy)carbonyl)--
L-arginyl)-L-lysinate
##STR00029##
[0137] A solution/suspension of H-Lys(Z)-OBzl hydrochloride (4.07
g, 10 mmol) and Z2-Arg(Z)--OH (5.77 g, 10 mmol) in anhydrous
dichloromethane (50 mL) under nitrogen was cooled on ice and
treated with HOBT hydrate (2.3 g, 15 mmol) and triethylamine (4.2
mL, 30 mmol), and the clear solution was stirred for 15 min EDC
hydrochloride (2.40 g, 12.5 mmol) was added, and the stirred
mixture allowed to warm to room temperature and stirred 20 h. The
above mixture was diluted with dichloromethane to 250 mL, washed
with 5% citric acid (100 mL), then water, saturated sodium
bicarbonate, and brine (50 mL each), dried (Na2SO4) and
concentrated in vacuo. The residual amorphous solid was triturated
from acetonitrile to afford 7.40 g (80%) as an amorphous white
solid. NMR (CDCl3): .quadrature..quadrature.9.41-9.28 (m, 3H),
7.38-7.19 (m, 25H), 6.87 (br s, 1H), 6.09-6.07 (m, 1H), 5.22 (s,
2H), 5.16-4.92 (m, 8H), 4.51-4.47 (m, 1H), 4.38-4.32 (m, 1H),
3.98-3.83 (m, 2H), 3.03-2.98 (m, 2H), 1.74-1.60 (m, 5H), 1.44-1.29
(m, 3H), 1.20-1.13 (m, 2H)
d. L-arginyl-L-lysine
##STR00030##
[0139] A stirred solution/suspension of benzyl
N6-((benzyloxy)carbonyl)-N2-(N2,Nd,Nw-tris((benzyloxy)carbonyl)-L-arginyl-
)-L-lysinate (6.97 g, 7.5 mmol) in methanol (125 mL) under nitrogen
was treated with 20% Pd(OH)2/C (0.50 g, Pearlman's catalyst), then
evacuated and purged several times with hydrogen via balloon. The
mixture was stirred for 48 h under hydrogen at 45.degree. C., then
the flask cooled and evacuated with nitrogen, diluted with water
(125 mL) and re-purged with hydrogen via balloon (gray precipitate
still present indicating incomplete deprotection). Stirring was
continued for 16 h more, after which only charcoal could be seen in
suspension. The flask was cooled and evacuated with nitrogen, then
the contents carefully filtered through Celite with water rinse.
The filtrate was concentrated in vacuo to afford 2.27 g (100%) as a
white foam. NMR (D.sub.2O): .delta. 3.97 (dd, 1H, J=8 Hz, 8 Hz),
3.85 and 3.24 (1H, both m, rotamers), 3.02-2.97 (m, 2H), 2.80-2.70
(m, 2H), 1.65-1.38 (m, 8H), 1.25-1.17 (m, 2H).
e.
Nd,Nw-bis((benzyloxy)carbonyl)-N2-(tert-butoxycarbonyl)-L-arginine
##STR00031##
[0141] A stirred solution of N.alpha.-Boc-arginine hydrochloride
hydrate (9.86 g, 30 mmol) in methanol (50 mL) was treated with 30%
sodium methoxide (5.20 g, 30 mmol), then stirred a few minutes and
filtered. The filtrate was concentrated in vacuo and the residual
foam was treated with toluene (3.times.30 mL) and three times
concentrated in vacuo to remove all methanol. The residual material
was suspended with stirring in anhydrous 1,2-dichloroethane (75 mL)
under nitrogen and treated with N,N-diisopropylethylamine (18.3 mL,
105 mmol). Chlorotrimethylsilane (13.4 mL, 105 mmol) was then added
at a rate to keep pot temperature below 35.degree. C., and the
mixture was heated to 40.degree. C. for 1.5 h, then cooled on ice.
Additional N,N-diisopropylethylamine (15.7 mL, 90 mmol) was added,
followed by benzyl chloroformate (12.85 mL, 90 mmol) in one
portion, and the mixture was stirred on ice for 15 min, allowed to
warm to room temperature over 30 min, and stirred at room
temperature for 4 h. The reaction mixture was plunged into 15%
citric acid (200 mL) and stirred for 15 min, then extracted with
dichloromethane (150 mL, then 2.times.50 mL). The combined organic
solution was washed with water and brine (100 mL each), dried
(Na.sub.2SO.sub.4), and concentrated in vacuo. The residual oil was
dissolved in dichloromethane and added to a silica gel column
(.about.600 cc) and eluted with 10% ethyl acetate/dichloromethane
(this gave 0.90 g of lactam byproduct), then 6%
ethanol/dichloromethane to afford (after solidification with a
small amount of ether and drying in vacuo) 8.11 g (50%) of as a
white solid. NMR (CDCl.sub.3): .delta. 9.43-9.28 (m, 2H), 7.39-7.26
(m, 10H), 5.28-5.26 (m, 1H), 5.22 (s, 2H), 5.12 (s, 2H), 4.28-4.24
(m, 1H), 3.99-3.95 (m, 2H), 1.82-1.63 (m, 4H), 1.39 (s, 9H).
f. benzyl
Nd,Nw-bis((benzyloxy)carbonyl)-N2-(N2,Nd,Nw-tris((benzyloxy)carb-
onyl)-L-arginyl)-L-argininate
##STR00032##
[0143] A solution/suspension of benzyl
N.sup..delta.,N.sup..omega.-bis((benzyloxy)carbonyl)-L-argininate
(6.64 g, 12.47 mmol) and Z2-Arg(Z)--OH (7.19 g, 12.47 mmol) in
anhydrous dichloromethane (60 mL) under nitrogen was cooled on ice
and treated with HOBT hydrate (2.87 g, 18.7 mmol) and triethylamine
(3.9 mL, 28 mmol), and the clear solution was stirred for 15 min
EDC hydrochloride (3.00 g, 15.65 mmol) was added, and the stirred
mixture allowed to warm to room temperature and stirred 20 h. The
above mixture was diluted with dichloromethane to 250 mL, washed
with 5% citric acid (100 mL), then water, saturated sodium
bicarbonate, and brine 75 mL each), dried (Na2SO4) and filtered.
The filtrate was added directly to a silica gel column (.about.400
cc) and eluted with 9:1 dichloromethane/ethyl acetate to afford
8.34 g (61%) as a white solid. NMR (CDCl.sub.3): .delta. 9.13 (br
s, 4H), 8.31 (d, 1H, J=7.5 Hz), 7.38-7.15 (m, 30H), 5.17-5.13 (m,
4H), 5.02-4.92 (m, 8H), 4.46 (d, 1H, J=5.5 Hz), 4.27-4.23 (m, 1H),
4.02-3.97 (m, 1H), 3.94-3.70 (m, 4H), 1.70-1.35 (m, 8H)
g. benzyl
Nd,Nw-bis((benzyloxy)carbonyl)-N2-(tert-butoxycarbonyl)-L-argini-
nate
##STR00033##
[0145] A cooled (0.degree. C.) stirred solution of
N.sup..delta.,N.sup..omega.-bis((benzyloxy)carbonyl)-N2-(tert-butoxycarbo-
nyl)-L-arginine (7.87 g, 14.5 mmol) and benzyl alcohol (2.28 mL, 22
mmol) in anhydrous DCM (125 mL) under nitrogen was treated with DCC
(3.30 g, 16 mmol) and DMAP (0.20 g, 1.64 mmol) and slowly allowed
to reach room temperature and stirred overnight (20 h). The
solution/suspension was filtered and the solid rinsed with DCM. The
filtrate was added directly to a column of silica gel (.about.300
cc) and eluted with 3% ethyl acetate/DCM to afford 9.01 g (98%) as
a white solid. NMR (CDCl.sub.3): .delta. 9.42 (br s, 1H), 9.22 (br
s, 1H), 7.38-7.24 (m, 15H), 5.18 (s, 2H), 5.13-5.07 (m, 5H),
4.32-4.28 (m, 1H), 3.94 (t, 2H, J=8 Hz), 1.78-1.59 (m, 4H), 1.39
(s, 9H)
h. benzyl Nd,Nw-bis((benzyloxy)carbonyl)-L-argininate
##STR00034##
[0147] A solution of benzyl
N.sup..delta.,N.omega.-bis((benzyloxy)carbonyl)-N2-(tert-butoxycarbonyl)--
L-argininate (8.86 g, 14 mmol) in 1:1 TFA/DCM (50 mL) was gently
warmed to induce bubbling, then stirred at room temperature for 1
h. The mixture was concentrated in vacuo (exhaustively to remove
all TFA possible) and the residual oil dissolved in 1:1 DCM/MeOH
and stirred for 15 min with DOWEX 550A-upw resin (.about.50 g) in
order to neutralize the salt. The mixture was filtered, resin
washed with 1:1 DCM/MeOH, and the filtrate concentrated in vacuo to
afford 6.64 g (89%) of benzyl
N.sup.8,N.sup..omega.-bis((benzyloxy)carbonyl)-L-argininate which
was carried forward without further purification.
i. L-arginyl-L-arginine
##STR00035##
[0149] A stirred solution/suspension of benzyl
N.sup..delta.,N.sup..omega.-bis((benzyloxy)carbonyl)-N2-(N2,Nd,Nw-tris((b-
enzyloxy)carbonyl)-L-arginyl)-L-argininate (2.182 g, 20 mmol) in
methanol (30 mL) and ethyl acetate (15 mL) under nitrogen was
treated with 10% Pd/C (0.15 g), then evacuated and purged several
times with hydrogen via balloon. The mixture was heated to
45.degree. C. and stirred for 24 h under hydrogen, then the flask
cooled and evacuated with nitrogen and carefully filtered through
Celite with water rinse. The filtrate was concentrated in vacuo to
afford 0.66 g (100%) as a white foam. NMR (D.sub.2O): .delta. 3.98
(dd, 1H, J=5 Hz, 8 Hz), 3.84 and 3.23 (1H, both m, rotamers)
3.01-2.97 (m, 4H), 1.65-1.38 (m, 8H).
Compound 1: (L,L) Lysyl-Lysine EPA Salt (Mono)
[0150] A stirred solution of (L,L)-lysyllisine (0.87 g, 3.16 mmol)
in 15 mL of methanol was heated to 45.degree. C., then treated with
a combined solution of EPA (1.0 g, 3.32 mmol) and
alpha-D-tocopherol (36 mg pre-dissolved in 0.5 mL of ethyl acetate
and added to the EPA solution) in methanol (15 mL). The reaction
cooled over 15 min and the solvent was concentrated in vacuo to a
foam. The foam was triturated from cold acetonitrile, collected and
dried to afford 1.56 g (86%) as a light pink solid. MP
92-94.degree. C. NMR (d.sub.4-MeOH): .delta. 5.25-5.42 (m, 10H),
4.23 (dd, 1H, J=5 Hz, 8 Hz), 3.40 (t, 1H, J=6.5 Hz), 2.78-2.91 (m,
12H), 2.16 (t, 2H, J=8 Hz), 2.03-2.12 (m, 4H), 1.89-1.80 (m, 1H),
1.73-1.58 (m, 9H), 1.50-1.40 (m, 4H), 0.95 (t, 3H, J=8 Hz).
.sup.13C NMR (151 MHz, cd.sub.3od) .delta. 181.05, 177.35, 173.99,
131.36, 129.42, 128.02, 128.00, 127.76, 127.73, 127.55, 127.50,
126.76, 54.32, 54.02, 47.16, 39.09, 38.98, 37.12, 33.73, 31.75,
26.84, 26.83, 26.81, 26.27, 25.15, 25.13, 25.12, 25.01, 22.33,
21.80, 20.09, 13.26.
Compound 2: (L,L) Lysyl-Lysine Bis-EPA Salt
[0151] A stirred solution of (L,L)-lysyllysine (549 mg, 2.0 mmol)
in methanol (10 mL) under nitrogen was heated to 45.degree. C.,
then treated with a combined solution of EPA (1.39 g, 4.6 mmol) and
alpha-D-tocopherol (50 mg pre-dissolved in 0.5 mL of ethyl acetate
and added to the EPA solution) in methanol (10 mL). The reaction
cooled over 15 min and the solvent was concentrated in vacuo to a
foam. The foam was triturated from cold acetonitrile, collected and
dried to afford 1.60 g (89%) as a very pale peach solid. MP
85-87.degree. C. NMR (d.sub.4-MeOH): .delta. 5.25-5.45 (m, 20H),
4.23 (dd, 1H, J=5 Hz, 8 Hz), 3.64 (t, 1H, J=6.5 Hz), 2.75-2.95 (m,
20H), 2.19 (t, 4H, J=8 Hz), 2.00-2.15 (m, 8H), 1.80-1.90 (m, 1H),
1.70-1.80 (m, 2H), 1.60-1.70 (m, 9H), 1.40-1.55 (m, 4H), 0.95 (t,
6H, J=8 Hz). .sup.13C NMR (151 MHz, cd.sub.3od) .delta. 179.49,
177.16, 171.11, 131.36, 129.18, 128.02, 127.97, 127.94, 127.74,
127.71, 127.59, 127.50, 126.76, 54.50, 53.25, 39.02, 38.81, 35.83,
32.11, 31.58, 26.66, 26.62, 26.58, 25.75, 25.15, 25.13, 25.12,
25.02, 22.37, 21.29, 20.09, 13.27.
Compound 3: (L,L) Lysyl-Lysine DHA Salt (Mono)
[0152] A stirred solution of (L,L)-lysyllisine (0.87 g, 3.17 mmol)
in methanol (13 mL) was heated to 45.degree. C., then treated with
a combined solution of DHA (1.09 g, 3.32 mmol) and
alpha-D-tocopherol (38 mg pre-dissolved in 0.5 mL of ethyl acetate
and added to the DHA solution)
in methanol (13 mL). The reaction cooled over 15 minutes and was
filtered through Celite. The filtrate was concentrated in vacuo to
a foam. The foam was triturated from cold acetonitrile, collected
and dried to afford 1.85 g (97%) as a white solid. MP 89-91.degree.
C. NMR (d.sub.4-MeOH): .delta. 5.25-5.44 (m, 12H), 4.23 (dd, 1H,
J=5 Hz, 8 Hz), 3.41 (t, 1H, J=6.5 Hz), 2.78-2.91 (m, 14H),
2.32-2.37 (m, 2H) 2.16-2.20 (m, 2H), 2.03-2.10 (m, 2H), 1.89-1.80
(m, 1H), 1.74-1.58 (m, 7H), 1.48-1.40 (m, 4H), 0.95 (t, 3H, J=7.5
Hz). .sup.13C NMR (151 MHz, CD.sub.3OD) .delta. 180.57, 177.35,
131.36, 129.20, 128.04, 128.02, 127.78, 127.76, 127.72, 127.69,
127.66, 127.59, 127.50, 126.76, 54.32, 53.97, 47.30, 47.16, 39.08,
38.97, 37.33, 33.64, 31.73, 26.81, 26.78, 25.15, 25.14, 25.11,
25.02, 23.99, 22.32, 21.77, 20.09, 13.27.
Compound (L,L) Lysyl-Lysine Bis DHA Salt
[0153] A stirred solution of (L,L)-lysyllisine (0.55 g, 2.0 mmol)
in methanol (15 mL) was heated to 45.degree. C., then treated with
a combined solution of DHA (1.51 g, 4.6 mmol) and
alpha-D-tocopherol (50 mg pre-dissolved in 0.5 mL of ethyl acetate
and added to the DHA solution) in methanol (15 mL). The reaction
cooled over 15 minutes and was filtered through Celite. The
filtrate was concentrated in vacuo to a foam. The foam was
triturated from cold acetonitrile, collected and dried to afford
1.68 g (87%) as a white solid. MP 64-67.degree. C. NMR
(d.sub.4-MeOH): .delta. 5.25-5.43 (m, 24H), 4.23 (dd, 1H, J=5 Hz, 8
Hz), 3.64 (t, 1H, J=6.5 Hz), 2.78-2.92 (m, 24H), 2.32-2.38 (m, 4H),
2.24-2.20 (m, 4H), 2.01-2.10 (m, 4H), 1.62-1.87 (m, 8H), 1.41-1.53
(m, 4H), 0.95 (t, 6H, J=7.5 Hz). .sup.13C NMR (151 MHz, cd.sub.3od)
.delta. 179.00, 177.15, 171.03, 131.36, 128.78, 128.02, 127.98,
127.95, 127.76, 127.73, 127.68, 127.63, 127.50, 126.76, 54.49,
53.24, 47.30, 47.16, 39.02, 38.80, 36.22, 32.07, 31.57, 26.65,
26.58, 25.15, 25.14, 25.11, 25.02, 23.54, 22.36, 21.26, 20.09,
Compound 5: (L,L) Lysyl-Lysine EPA DHA Salt
[0154] A stirred solution of (L,L)-lysyllisine (0.53 g, 1.93 mmol)
in 15 mL of methanol was heated to 45.degree. C., then treated with
a combined solution of EPA (0.60 g, 1.97 mmol) and DHA (0.65 g,
1.97 mmol) in 15 mL of methanol. The reaction cooled over 15
minutes and was filtered through Celite. The filtrate was
concentrated in vacuo to give an off-white foam. The foam was
triturated from cold acetonitrile, collected and dried to afford
1.56 g (89%) as a beige solid. MP 76-78.degree. C. NMR
(d.sub.4-MeOH): .delta. 5.25-5.43 (m, 22H), 4.23 (dd, 1H, J=5 Hz, 8
Hz), 3.60 (t, 1H, J=6.5 Hz), 2.78-2.92 (m, 22H), 2.32-2.37 (m, 2H),
2.18-2.23 (m, 4H), 2.03-2.12 (m, 6H), 1.60-1.88 (m, 10H), 1.41-1.52
(m, 4H), 0.95 (t, 6H, J=7.5 Hz). .sup.13C NMR (151 MHz, cd.sub.3od)
.delta. 179.58, 179.23, 177.17, 171.18, 131.36, 129.19, 128.85,
128.02, 127.96, 127.95, 127.93, 127.76, 127.74, 127.73, 127.71,
127.68, 127.62, 127.59, 127.50, 126.76, 54.50, 53.27, 47.59, 47.45,
47.30, 47.16, 39.03, 38.81, 36.42, 35.87, 32.15, 31.58, 26.67,
26.63, 26.59, 25.76, 25.15, 25.14, 25.12, 25.11, 25.02, 23.62,
22.37, 21.30, 20.09, 13.27.
Compound 6: (L,L) Lysyl-Lysine Bis Gabapentin Salt
[0155] A stirred solution of (L,L)-lysyllisine (0.154 g, 0.56 mmol)
in 8 mL of methanol was heated to 45.degree. C., then gabapentin
(0.20 g, 1.18 mmol) was added. The reaction cooled over 15 minutes
and was filtered through Celite. The filtrate was concentrated in
vacuo to give a foam. The foam was triturated from cold
acetonitrile, collected and dried to afford 0.30 g (92%) as a white
solid. MP 132-134.degree. C. NMR (D.sub.2O): .delta. 4.01 (dd, 1H,
J=5 Hz, 8 Hz), 3.24 (t, 1H, J=6.5), 2.78-2.74 (m, 8H), 2.22 (s,
4H), 1.67-1.43 (m, 8H), 1.31-1.17 (m, 24H).
Compound 7: (L,L) Lysyl-Lysine EPA Niacin Salt
[0156] A stirred solution of (L,L)-lysyllisine (0.12 g, 0.44 mmol)
in 5 mL of methanol was heated to 45.degree. C., then Niacin (0.05
g, 0.44 mmol) was added. After 10 minutes a combined solution of
EPA (0.39 g, 1.31 mmol) and alpha-D-tocopherol (6 mg pre-dissolved
in 0.5 mL of ethyl acetate and added to the EPA solution) in 5 mL
of methanol was added. The reaction cooled over 15 minutes and the
solvent was concentrated in vacuo to give a foam. The foam was
triturated from cold acetonitrile, collected and dried to afford
0.25 g (82%) as an off-white solid. MP 116-119.degree. C. NMR
(d.sub.4-MeOH): .delta. 9.04 (s, 1H), 8.53 (dd, 1H, J=2 Hz, 5 Hz),
8.30 (dt, 1H, J=2 Hz, 8 Hz), 7.43 (dd, 1H, J=5 Hz, 8 Hz), 5.41-5.25
(m, 10H), 4.24 (dd, 1H, J=5 Hz, 8 Hz), 3.75 (t, 1H, J=6.5 Hz),
2.94-2.78 (m, 12H), 2.22 (t, 2H, J=7.5 Hz), 2.13-2.07 (m, 4H),
1.89-1.79 (m, 4H), 1.73-1.60 (m, 6H), 1.59-1.43 (m, 4H), 0.95 (t,
3H, J=7.5 Hz)
Compound 8: (L,L) Lysyl-Lysine EPA Ibuprofen Salt
[0157] A solution of (L,L)-lysyllisine (0.15 g, 0.56 mmol) in 5 mL
of methanol was treated with Ibuprofen (0.12 g, 0.56 mmol) and
stirred for 5 minutes. The solvent was concentrated in vacuo to
give a foam. The foam was triturated in acetonitrile and the
solvent decanted and replaced with 5 mL of methanol. The solution
was heated to 45.degree. C., then treated with a combined solution
of EPA (0.20 g, 0.67 mmol) and alpha-D-tocopherol (15 mg
pre-dissolved in 0.5 mL of ethyl acetate and added to the EPA
solution) in 3 mL of methanol. The reaction cooled over 15 minutes
and the solvent was concentrated in vacuo to give a foam. The foam
was triturated from cold acetonitrile, collected and dried to
afford 0.31 g (70%) as a beige solid. MP 89-92.degree. C. NMR
(d.sub.4-MeOH): .delta. 7.22 (d, 2H, J=6 Hz), 7.03 (d, 2H, J=6 Hz),
5.41-5.25 (m, 10H), 4.23 (dd, 1H, J=5 Hz, 8 Hz), 3.62 (t, 1H, J=6.5
Hz), 3.54 (dd, 1H, J=7 Hz, 14 Hz), 2.90-2.78 (m, 12H), 2.41 (d, 2H,
J=7 Hz), 2.20 (t, 2H, J=7.5 Hz), 2.13-2.05 (m, 4H), 1.89-1.58 (m,
11H), 1.51-1.42 (m, 4H), 1.38 (d, 3H, J=7 Hz), 0.95 (t, 3H, J=7.5
Hz), 0.86 (d, 6H, J=6.5 Hz)
##STR00036##
Compound 9: (L,L) Lysyl-Lysine EPA Methanesulfonic Acid Salt
[0158] A stirred solution of (L,L)-lysyllisine (0.09 g, 0.33 mmol)
in 5 mL of methanol was heated to 45.degree. C., then
methanesulfonic acid (0.03 g, 0.33 mmol) was added. After 10
minutes a combined solution of EPA (0.297 g, 0.98 mmol) and
alpha-D-tocopherol (65 mg pre-dissolved in 0.5 mL of ethyl acetate
and added to the EPA solution) in 5 mL of methanol was added. The
reaction cooled over 15 minutes and was filtered through Celite.
The filtrate was concentrated in vacuo to give a foam. The foam was
triturated from cold acetonitrile, collected and dried to afford
0.19 g (84%) as a beige solid. NMR (d.sub.4-MeOH): .delta.
5.41-5.25 (m, 10H), 4.23 (dd, 1H, J=5 Hz, 8 Hz), 3.68 (t, 1H, J=6.5
Hz), 2.95-2.88 (m, 4H), 2.85-2.79 (m, 8H), 2.69 (s, 3H), 2.20 (t,
2H, J=7.5 Hz), 2.11-2.03 (m, 4H), 1.90-1.73 (m, 4H), 1.71-1.60 (m,
6H), 1.54-1.42 (m, 4H), 0.95 (t, 3H, J=7.5 Hz).
Compound 10: (L,L) Arginyl-Lysine Bis Gabapentin Salt
[0159] A stirred mixture of (L,L)-arginyllysine (151 mg, 0.5 mmol)
and gabapentin (180 mg, 1.05 mmol) in 2:1 methanol/water (6 mL) was
heated to 45.degree. C. The reaction cooled over 15 minutes and was
concentrated in vacuo to a foam. The foam was triturated from cold
acetonitrile, collected and dried to afford 294 mg (91%) as a white
solid. NMR (DMSO): .delta. 3.92 (dd, 1H, J=8 Hz, 8 Hz), 3.21-3.15
(m, 1H) 3.01-2.95 (m, 2H), 2.67 (s, 4H), 2.58 (t, 2H, J=7 Hz), 2.17
(s, 4H), 1.92 (s, 2H), 1.61-1.10 (m, 28H).
Compound 11: (L,L) Arginyl-Arginine Bis Gabapentin Salt
[0160] A stirred mixture of L-arginyl-L-arginine (330 mg, 1.0 mmol)
and gabapentin (360 mg, 2.1 mmol) in 2:1 methanol/water (12 mL) was
heated to 45.degree. C. The reaction cooled for 15 minutes and was
filtered through Celite. The filtrate was concentrated in vacuo to
a foam. The foam was triturated from cold acetonitrile, collected
and dried to afford 602 mg (87%) as a white solid. NMR (D2O):
.delta. 4.00 (dd, 1H, J=5 Hz, 8 Hz), 3.26-3.23 (m, 1H), 3.03-2.98
(m. 4H), 2.74 (s, 4H), 2.19 (s, 4H), 1.56-1.14 (m, 28H).
Stability
[0161] Stability analysis was performed using 1H NMR (400 MHz) with
acetic acid-d4 at various time point (typically days 1, 14, 30 and
60) using samples exposed to air and light at ambient temperature
(clear vials with no caps). Semi-quantitative analysis based on
integration and resolution of the vinyl peaks (2.75-3.00 ppm) was
used to categorize samples as 1=>95%, 2=75-95%, 3=50-75%,
4=<50%, where the percent value indicate the amount of sample
remaining relative to the original sample at day 1.
TABLE-US-00003 TABLE 3 Stability.sup.1 of representative compounds
compared to EPA ethyl ester Compound Day 3 Day 14 Day 30 Day 60 EPA
ethyl ester 1 1-2 4 4 Lysyl-lysine EPA 1 1 1 1 Lysyl-lysine Bis-EPA
1 1 1 1 Arginyl-arginine Bis-EPA 1 1 1 1 Arginyl-lysine Bis-EPA 1 1
1 1 Lysyl-lysine DHA 1 1 1 1 Lysyl-lysine Bis DHA 1 1 1 1
Arginyl-arginine Bis-DHA 1 1 1 1 Arginyl-lysine Bis-DHA 1 1 1 1
Lysyl-lysine EPA Nicotinic Acid 1 1 1 1 Lysyl-lysine EPA Ibuprofen
1 1 1 1 .sup.1Stability by NMR of compounds exposed to air for the
indicated time: 1 = >95%, 2 = 75-95%, 3 = 50-75%, 4 =
<50%.
Pharmacokinetic Properties
[0162] Experiments Via Oral Gavage:
[0163] Oral pharmacokinetic parameters of TP-212 (Lysyl-Lysine
Bis-EPA), TP-312 (Lysyl-Lysine Bis-DHA), EPA-EE, EPA FFA and DHA
FFA were determined in Sprague-Dawley rats. EPA-EE is 97% pure
ethyl ester form of eicosapentaenoic acid. It is the principal
polyunsaturated fatty acid in Vascepa, an Omega-3 compound approved
in the U.S. for the treatment of severe hypertriglyceridemia. EPA
FFA is 97% pure free fatty acid form of eicosapentaenoic acid and
DHA FFA is 98% pure free fatty acid DHA. EPA FFA and DHA FFA are
the two principal poly-unsaturated fatty acids in Epanova, an
Omega-3 compound approved in the U.S. for the treatment of severe
hypertriglyceridemia. The subject drugs were administered by oral
gavage as an aqueous solution in de-ionized water to 6
Sprague-Dawley rats per group (3 males and 3 females). Rats were
dosed at 40 mg/kg. Blood samples were obtained from each rat by
jugular vein catheter. Samples were collected at pre-dose and at
0.25, 0.5, 1, 2, 4, 8, 12 and 18 hours post dose. Blood samples
were centrifuged to separate red blood cells and the resulting
plasma samples were analyzed for free EPA or free DHA. Free EPA or
free EPA was extracted from rat plasma with hexane at acidic
condition without hydrolysis. Detection was by MS-MS monitoring of
negative ion for EPA or DHA. Calculated pharmacokinetic values are
mean values from 6 rats for each study drug.
[0164] Experiments Via Intravenous Injection:
[0165] TP-212 and TP-312 were administered by intravenous injection
(20 mg/kg) to Sprague-Dawley rats (n=6/group--3 male and 3 female),
and plasma was obtained from serial blood samples taken from each
animal at scheduled times (Predose, 0.25, 0.5, 1, 2, and 4 hours
post dose). Blood samples were centrifuged to separate red blood
cells and the resulting plasma samples were analyzed for free EPA
or free DHA. Free EPA or free DHA was extracted from rat plasma
with hexane at acidic condition without hydrolysis. Detection was
by MS-MS monitoring of negative ion for EPA or DHA. Calculated
pharmacokinetic values are mean values from 6 rats for each study
drug.
[0166] For all figures and tables, the data are adjusted as follows
for comparative purposes. First, the actual plasma levels are
baseline adjusted. Thus, for each study arm, the pre-dose level
(time=0) is subtracted from the actual plasma levels at each time
point, with negative values adjusted to zero. Second, these
baseline adjusted values are further adjusted to be molar dose
equivalent on a comparative basis among the study arms with respect
to EPA or DHA payload (i.e., as if equal amounts of free fatty acid
EPA or DHA are administered in each study arm).
Results
[0167] FIG. 1 and Table 4 show the plasma levels of free EPA of
TP-212 compared to EPA-EE and EPA-FFA, all administered via oral
gavage. The data indicate that the Cmax of TP-212 is approximately
8.9 and 7.4 times greater than EPA-EE and EPA-FFA, respectively;
and the AUC over 18 hours (ug*h/mL) of TP-212 is approximately 2.6
and 1.8 times greater than EPA-EE and EPA-FFA, respectively.
TABLE-US-00004 TABLE 4 Plasma levels of free EPA of TP-212 (oral).
ug/mL * Hour TP-212 EPA-EE EPA-FFA 0 0.0 0.0 0.0 0.25 0.3 0.3 0.0
0.5 9.5 0.5 1.3 1 3.9 0.7 0.5 2 3.2 0.8 0.8 4 1.5 1.1 1.1 8 0.6 0.5
0.9 12 0.0 0.0 0.3 18 0.0 0.0 0.0 Tmax 30 min 4 hrs 30 min Cmax
(ug/mL) 9.5 1.1 1.3 AUC [0-18 hrs (ug*h/mL)] 18.3 7.2 10.5 * Molar
dose equivalent, baseline adjusted.
[0168] FIG. 2 and Table 5 show the plasma levels of free DHA of
TP-312 compared to DHA FFA, all administered via oral gavage. The
data indicate that the Cmax of TP-312 is approximately 3.0 times
greater than DHA-FFA; and the AUC over 18 hours (ug*h/mL) of TP-312
is approximately 1.2 times greater than DHA-FFA.
TABLE-US-00005 TABLE 5 Plasma levels of free DHA of TP-312 (oral).
ug/mL * Hour TP-312 DHA-FFA 0 0.0 0.0 0.25 0.0 0.0 0.5 4.6 1.8 1
6.3 1.8 2 10.4 2.2 4 5.6 3.4 8 1.4 1.7 12 0.0 2.1 18 0.0 1.2 Tmax 2
hrs 4 hrs Cmax (ug/mL) 10.4 3.4 AUC [0-18 hrs (ug*h/mL)] 44.4 36.3
* Molar Dose Equivalent, Baseline Adjusted
[0169] FIG. 3 and Table 6 show the plasma levels of free EPA of
TP-212 administered via intravenous injection (TP-212iv) versus
TP-212 administered via oral gavage (TP-212po) versus EPA-FFA
administered via oral gavage (EPApo). The data indicate that the
Cmax of TP-212iv is approximately 3.6 and 27 times greater than
TP-212po and EPA-FFApo, respectively; the AUC over 1 hour of
TP-212iv is approximately 1.8 and 13.4 times greater than TP-212po
and EPApo, respectively; and the AUC over 4 hours of TP-212iv is
approximately 1.2 and 5.1 times greater than TP-212po and EPApo,
respectively.
TABLE-US-00006 TABLE 6 Plasma levels of free EPA of TP-212 (iv)
ug/mL * Min TP-212iv TP-212po EPApo 0 0.0 0.0 0.0 1 34.5 ** ** 5
6.6 ** ** 10 6.4 ** ** 15 11.4 0.3 0.0 30 8.8 9.5 1.3 60 4.6 3.9
0.5 120 2.3 3.2 0.8 240 1.8 1.5 1.1 Tmax 1 min 30 min 30 min Cmax
(ug/mL) 34.5 9.5 1.3 AUC [0-1 hrs (ug*h/mL)] 8.3 4.6 0.6 AUC [0-4
hrs (ug*h/mL)] 15.9 12.8 3.1 * Molar dose equivalent, baseline
adjusted ** No blood samples on studies with oral doses
[0170] FIG. 4 and Table 7 show the plasma levels of free DHA of
TP-312 administered via intravenous injection (TP-312iv) versus
TP-312 administered via oral gavage (TP-312po) versus DHA-FFA
administered via oral gavage (DHApo). The data indicate that the
Cmax of TP-312iv is approximately 6.4 and 19.3 times greater than
TP-312po and DHApo, respectively; the AUC over 1 hour of TP-312iv
is approximately 4.9 and 14.7 times greater than TP-312po and
DHApo, respectively; and the AUC over 4 hours of TP-312iv is
approximately 1.6 times and 5.0 times greater than TP-312po and
DHApo, respectively.
TABLE-US-00007 TABLE 7 Plasma levels of free DHA of TP-312 (iv).
ug/mL * Min TP-312iv TP-312po DHApo -- 0.0 0.0 0.0 1 66.4 ** ** 5
15.1 ** ** 10 16.6 ** ** 15 17.5 0.0 0.0 30 16.4 4.6 1.8 60 12.9
6.3 1.8 120 8.5 10.4 2.2 240 7.8 5.6 3.4 Tmax 1 min 2 hrs 4 hrs
Cmax (ug/mL) 66.4 10.4 3.4 AUC [0-1 hrs (ug*h/mL)] 16.3 3.3 1.1 AUC
[0-4 hrs (ug*h/mL)] 43.3 27.6 8.7 * Molar dose equivalent, baseline
adjusted ** No blood samples on studies with oral doses
Ophthalmic Formulations
[0171] In an embodiment where the ophthalmic formulation is an
aqueous formulation, it has been determined in mice that certain
formulations can be administered to the surface of the eye without
producing irritation that would make them unsuitable for such
application. Irritation parameters of TP-211-1, TP-212-1, TP-311-1
and TP-312-1 related to hyperemia, squinting, and discharge were
determined in Balb/C mice. Mice were dosed topically to the cornea
using a calibrated micropipette, with a 3 .mu.L drop of treatment
in each eye four times daily for 3 consecutive days at various
concentrations.
[0172] TP-211-1 is Lysyl-lysine EPA. TP-212-1 is Lysyl-lysine
bis-EPA. TP-311-1 is Lysyl-lysine DHA. TP-312-1 is Lysyl-lysine
bis-DHA. The study drugs were prepared using phosphate buffered
saline (PBS) to reach target concentrations ranging from 0.1% to
1.0%.
[0173] The animals were monitored for scratching at the eye or any
other abnormal behavior post-dose. Ocular exams were conducted on
Days 1 and 3 with use of a Micron III camera system to take high
resolution videos of the right eye of each animal. Clinical signs
including but not limited to hyperemia, discharge, lid swelling,
and squinting were evaluated by trained personnel who was masked as
to group assignment. Each clinical sign was graded on a scale of
0-4, with the exception of lid swelling which was graded on a scale
of 0-2. Scores were averaged and are expressed as the mean.+-.SEM
for each group. Statistical significance was assessed between the
treatment arms and the vehicle group. Any significance is noted by
asterisks (*=p<0.05, **=p<0.01, ***=p<0.001).
Summary of Observations
[0174] TP-211-1: TP-211-1 was tested at 0.5%, 0.25%, and 0.1%
concentrations. The 0.5% concentration caused some initial increase
in hyperemia, which returned to baseline levels by the last dose on
Day 3. The other concentrations maintained baseline levels of
hyperemia throughout both evaluations. There were no statistically
significant changes in squinting for this test article. Therefore,
TP-211-1 is not irritating at concentrations of less than 0.5%.
[0175] TP-311-1: TP-311-1 was tested at 1.0% and 0.5%
concentrations. The 1.0% group had elevated hyperemia after the
first dose administration with no increase in squinting levels. The
lowest concentration of 0.5% did not cause any increase in
hyperemia or squinting throughout. Therefore, TP-311-1-1 is not
irritating at concentrations of less than 1.0%.
[0176] TP-212-1: TP-212-1 was tested at 1.0% and 0.5%
concentrations. The 1.0% and 0.5% concentrations showed an increase
in hyperemia (but not squinting) after the first dose. However,
this initial increase in hyperemia returned to baseline levels by
the final evaluation. This indicates that concentrations of less
than 1.0% of TP-212-1 may cause some initial irritation upon the
very first installation, but the animals acclimated well to repeat
dosing.
[0177] TP-312-1: TP-312-1 was tested at 1.0%, 0.5% and 0.1%
concentrations. The 1.0% and 0.5% concentrations caused some
initial hyperemia after the first dose. The 0.1% concentration did
not cause any significant increase in hyperemia after the first
dose. All concentrations remained at baseline levels of hyperemia
by the last dose of the day, and none of the concentrations caused
squinting throughout the study. Therefore, TP-312-1 is well
tolerated in mice at a concentration of less than 1.0%.
[0178] Summary: All concentrations noted above appeared comfortable
by Day 3 of dosing.
[0179] FIGS. 5A-8C show each endpoint for one of the test
compounds, TP-312-1. Comparable data was compiled for the other
test articles. In each figure, FIG. 5A, FIG. 6A, FIG. 7A, and FIG.
8A are the pre-dose/baseline, FIG. 5B, FIG. 6B, FIG. 7B, and FIG.
8B are 30 minutes post dose, and FIG. 5C, FIG. 6C, FIG. 7C, and
FIG. 8C are the delta of FIG. 5A, FIG. 6A, FIG. 7A, and FIG. 8A,
respectively, and FIG. 5B, FIG. 6B, FIG. 7B, and FIG. 8B,
respectively.
EQUIVALENTS
[0180] Those skilled in the art will recognize or be able to
ascertain using no more than routine experimentation, many
equivalents to the specific embodiments of the invention described
herein. Such equivalents are intended to be encompassed by the
following claims.
[0181] All references cited herein are incorporated herein by
reference in their entirety and for all purposes to the same extent
as if each individual publication or patent or patent application
was specifically and individually indicated to be incorporated by
reference in its entirety for all purposes.
[0182] The present invention is not to be limited in scope by the
specific embodiments described herein. Indeed, various
modifications of the invention in addition to those described
herein will become apparent to those skilled in the art from the
foregoing description and accompanying figures. Such modifications
are intended to fall within the scope of the appended claims.
* * * * *