U.S. patent application number 10/521492 was filed with the patent office on 2006-05-18 for modified amino acid for the inhibition of platelet aggregation.
Invention is credited to Moise Azria, Simon David Bateman.
Application Number | 20060106110 10/521492 |
Document ID | / |
Family ID | 30116066 |
Filed Date | 2006-05-18 |
United States Patent
Application |
20060106110 |
Kind Code |
A1 |
Bateman; Simon David ; et
al. |
May 18, 2006 |
Modified amino acid for the inhibition of platelet aggregation
Abstract
A method of inhibiting blood platelet aggregation in a mammal is
provided. The method comprises the administration of a platelet
aggregation inhibiting amount of a modified amino acid or
pharmaceutically acceptable salt thereof.
Inventors: |
Bateman; Simon David;
(RANDOLPH, NJ) ; Azria; Moise; (Basel,
CH) |
Correspondence
Address: |
NOVARTIS;CORPORATE INTELLECTUAL PROPERTY
ONE HEALTH PLAZA 104/3
EAST HANOVER
NJ
07936-1080
US
|
Family ID: |
30116066 |
Appl. No.: |
10/521492 |
Filed: |
July 16, 2003 |
PCT Filed: |
July 16, 2003 |
PCT NO: |
PCT/EP03/07739 |
371 Date: |
August 23, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60396898 |
Jul 17, 2002 |
|
|
|
Current U.S.
Class: |
514/563 ;
514/11.8; 514/11.9; 514/13.8; 514/5.9; 514/56 |
Current CPC
Class: |
A61K 31/727 20130101;
A61K 38/23 20130101; A61K 38/29 20130101; A61K 38/28 20130101; A61K
2300/00 20130101; A61K 38/28 20130101; A61K 31/166 20130101; A61K
38/29 20130101; A61P 7/02 20180101; A61K 38/23 20130101; A61K
31/198 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
514/563 ;
514/003; 514/012; 514/056 |
International
Class: |
A61K 31/198 20060101
A61K031/198; A61K 38/28 20060101 A61K038/28; A61K 38/29 20060101
A61K038/29; A61K 31/727 20060101 A61K031/727 |
Claims
1-13. (canceled)
14. A method of inhibiting platelet aggregation in a mammal
comprising administering a platelet inhibiting amount of a modified
amino acid, or a pharmaceutically acceptable salt thereof.
15. The method of claim 14, wherein the modified amino acid is
N-(-5-chlorosalicyloyl)-8-aminocaprylic acid (5-CNAC), or a
pharmaceutically acceptable salt thereof.
16. The method of claim 14, wherein the modified amino acid is
present in an amount of about 25 mg to about 400 mg.
17. The method of claim 14, wherein the modified amino acid is
present in an amount of about 100 mg to about 200 mg.
18. The method of claim 14, wherein the method further comprises
administering a pharmacologically active agent with said modified
amino acid, or pharmaceutically acceptable salt thereof, wherein
the modified amino acid or salt thereof is present in an amount
effective to inhibit platelet aggregation.
19. The method of claim 18, wherein the modified amino acid is
N-(-5-chlorosalicyloyl)-8-aminocaprylic acid (5-CNAC), or a
pharmaceutically acceptable salt thereof.
20. The method of claim 19, wherein the pharmacologically active
agent comprises heparin, insulin, parathyroid hormone or
calcitonin.
21. The method of claim 20, wherein the mammal is human and the
calcitonin is salmon calcitonin.
22. The method of claim 18, wherein the pharmacologically active
agent is present in an amount of 0.05% to 70% by weight relative to
the total weight of the pharmaceutical composition.
23. The method of claim 18 wherein the pharmaceutical composition
comprises calcitonin and 5-CNAC or a pharmaceutically acceptable
salt thereof.
24. A pharmaceutical composition comprising a platelet aggregation
inhibiting amount of a modified amino acid, or a pharmaceutically
acceptable salt thereof.
25. The composition of claim 24, further comprising a
pharmacologically active agent and a modified amino acid, or
pharmaceutically acceptable salt thereof, wherein the modified
amino acid or salt thereof is present in an amount effective to
inhibit platelet aggregation.
Description
[0001] Platelet activation and aggregation are involved in unstable
angina and acute myocardial infarction, in reocclusion following
thrombolytic therapy and angioplasty, in transient ischemic attacks
and in a variety of other cardiovascular disorders. When a blood
vessel is damaged either by acute intervention, such as
angioplasty, or more chronically by the pathophysiological
processes of atherosclerosis, platelets are activated to adhere to
the damaged surface and to each other. This platelet activation,
adherence and aggregation may lead to occlusive thrombus formation
in the lumen of the blood.
[0002] Various agents have been studied for many years as potential
targets for inhibiting platelet aggregation and thrombus formation.
For example, aspirin has come into use as a prophylactic
antithrombotic agent due its ability to inhibit platelet
aggregation.
[0003] U.S. Pat. Nos. 5,773,647 ('647) and 5,866,536 ('536)
describe compositions for the oral delivery of pharmacologically
active agents with modified amino acids, such as
N-(5-chlorosalicyloyl)-8-aminocaprylic acid (5-CNAC),
N-(10-[2-hydroxybenzoyl]aminodecanoic acid (SNAD), and
N-(8-[2-hydroxybenzoyl]amino)caprylic acid (SNAC). In addition, WO
00/059863 ('863) discloses the disodium salts of formula I ##STR1##
wherein [0004] R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are
independently hydrogen, --OH, --NR.sup.6R.sup.7, halogen,
C.sub.1-C.sub.4alkyl or C.sub.1-C.sub.4alkoxy; [0005] R.sup.5 is a
substituted or unsubstituted C.sub.2-C.sub.16alkylene, substituted
or unsubstituted C.sub.2-C.sub.16alkenylene, substituted or
unsubstituted C.sub.1-C.sub.12alkyl(arylene), or substituted or
unsubstituted aryl(C.sub.1-C.sub.12alkylene); and [0006] R.sup.6
and R.sup.7 are independently hydrogen, oxygen or
C.sub.1-C.sub.4alkyl; and hydrates and solvates thereof as
particularly efficacious for the oral delivery of active
agents.
[0007] Surprisingly, it has now been discovered that the modified
amino acids of '647, '536 and '863 are effective inhibitors of
blood platelet aggregation. Thus, pharmaceutical compositions
employing the modified amino acids of '647, '536 and '863 as
carriers for pharmacologically active agents have the added
advantage of inhibiting blood platelet aggregation.
[0008] Accordingly, the present invention provides a method of
inhibiting platelet aggregation in a mammal, preferably human,
comprising the administration of a platelet aggregation inhibiting
amount of a modified amino acid, or a pharmaceutically acceptable
salt thereof.
[0009] In another embodiment, the invention provides a method of
inhibiting platelet aggregation in a mammal, preferably human,
comprising the administration of a pharmaceutical composition
comprising a platelet aggregation inhibiting amount of a modified
amino acid or pharmaceutically acceptable salt thereof.
[0010] In an additional embodiment, the present invention provides
a method of inhibiting platelet aggregation in a mammal, preferably
human, receiving a pharmacologically active agent comprising the
administration of a pharmaceutical composition comprising said
pharmacologically active agent and a modified amino acid or a
pharmaceutically acceptable salt thereof, wherein the modified
amino acid or salt thereof is present in an amount effective to
inhibit platelet aggregation.
[0011] The invention is furthermore concerned with a method of
inhibiting platelet aggregation in a mammal (preferably human)
comprising administering a platelet aggregation inhibiting amount
of N-(-5-chlorosalicyloyl)-8-aminocaprylic acid (5-CNAC), or a
pharmaceutically acceptable salt thereof, to said patient.
[0012] In an additional embodiment, the invention provides a method
of inhibiting platelet aggregation in a mammal, preferably human,
comprising the administration of a pharmaceutical composition
comprising a platelet aggregation inhibiting amount of 5-CNAC or
pharmaceutically acceptable salt thereof.
[0013] In a yet further embodiment, the invention provides a method
of inhibiting platelet aggregation in a mammal, preferably human,
receiving a pharmacologically active agent comprising the
administration of a pharmaceutical composition comprising said
pharmacologically active agent and 5-CNAC or pharmaceutically
acceptable salt thereof, wherein the 5-CNAC or salt thereof is
present in an amount effective to inhibit platelet aggregation.
[0014] It is to be understood that in embodiments of the invention
comprising both a pharmacologically active agent and a modified
amino acid, the platelet aggregation inhibition activity is a
function of the modified amino acid. Such platelet aggregation
activity is not a function of the pharmacologically active
agent.
[0015] In an other embodiment, the invention provides a method of
inhibiting platelet aggregation in a mammal receiving heparin,
insulin, parathyroid hormone or calcitonin treatment, comprising
administering a pharmaceutical composition comprising said heparin,
insulin, parathyroid hormone or calcitonin and a modified amino
acid, or pharmaceutically acceptable salt thereof, wherein the
modified amino acid is present in an amount effective to inhibit
platelet aggregation.
[0016] In an other embodiment, the invention provides a method of
inhibiting platelet aggregation according to the invention, wherein
the calcitonin is salmon calcitonin.
[0017] In an other embodiment, the invention provides a method of
inhibiting platelet aggregation according to the invention, wherein
the modified amino acid is present in an amount of about 25 mg to
about 400 mg preferably in an amount of about 100 mg to about 200
mg.
[0018] In a further embodiment, the invention provides a method of
inhibiting platelet aggregation according to the invention, wherein
the pharmacologically active agent is present in an amount of 0.05%
to 70% by weight relative to the total weight of the pharmaceutical
composition.
[0019] In a further embodiment, the invention provides a method of
inhibiting platelet aggregation in a mammal receiving heparin,
insulin, parathyroid hormone or calcitonin treatment, comprising
administering a pharmaceutical composition comprising said heparin,
insulin, parathyroid hormone or calcitonin and 5-CNAC or
pharmaceutically acceptable salt thereof, wherein the 5-CNAC is
present in an amount effective to inhibit platelet aggregation.
[0020] In a further embodiment, the invention provides a method of
inhibiting platelet aggregation according to the invention, wherein
the pharmaceutical composition comprises calcitonin and 5-CNAC or a
pharmaceutically acceptable salt thereof, and said mammal is
human.
[0021] In a further embodiment, the invention provides a method of
inhibiting platelet aggregation according to the invention, wherein
the calcitonin is salmon calcitonin.
[0022] The present invention is directed to the use of a
pharmaceutical composition comprising a platelet aggregation
inhibiting amount of a modified amino acid, or a pharmaceutically
acceptable salt thereof, for the manufacture of a medicament for
the inhibition of platelet aggregation.
[0023] The invention also concerns the use of a pharmaceutical
composition comprising a pharmacologically active agent and a
modified amino acid, or pharmaceutically acceptable salt thereof,
wherein the modified amino acid or salt thereof is present in an
amount effective to inhibit platelet aggregation, for the
manufacture of a medicament for the inhibition of platelet
aggregation.
[0024] In a preferred embodiment the invention concerns the use of
a pharmaceutical composition comprising a pharmacologically active
agent and a modified amino acid, or pharmaceutically acceptable
salt thereof, wherein the modified amino acid or salt thereof is
present in an amount effective to inhibit platelet aggregation for
the manufacture of a medicament for the inhibition of platelet
aggregation said modified amino acid being
N-(-5-chlorosalicyloyl)-8-aminocaprylic acid (5-CNAC), or a
pharmaceutically acceptable salt thereof.
[0025] The invention also concerns the use of a pharmaceutical
composition comprising heparin, insulin, parathyroid hormone or
calcitonin and a modified amino acid, or pharmaceutically
acceptable salt thereof, wherein the modified amino acid or salt
thereof is present in an amount effective to inhibit platelet
aggregation for the manufacture of a medicament for the inhibition
of platelet aggregation in a mammal (preferably human) receiving
heparin, insulin, parathyroid hormone (PTH) or calcitonin
treatment.
[0026] In a preferred embodiment the invention concerns the use of
a pharmaceutical composition according to the invention, wherein
the calcitonin is salmon calcitonin.
[0027] In an other embodiment the invention concerns the use of a
pharmaceutical composition according to the invention, wherein the
modified amino acid is present in an amount of about 25 mg to about
400 mg.
[0028] In a preferred embodiment the invention concerns the use of
a pharmaceutical composition according to the invention wherein the
modified amino acid is present in an amount of about 100 mg to
about 200 mg.
[0029] In an other preferred embodiment the invention concerns the
use of a pharmaceutical composition according to the invention,
wherein the pharmacologically active agent is present in an amount
of 0.05% to 70% by weight relative to the total weight of the
pharmaceutical composition.
[0030] The invention is also directed to the use of a
pharmaceutical composition according to the invention wherein the
pharmaceutical composition comprises calcitonin and 5-CNAC or a
pharmaceutically acceptable salt thereof, and said mammal is
human.
[0031] The invention is also directed to a pharmaceutical
composition comprising a platelet aggregation inhibiting amount of
a modified amino acid, or a pharmaceutically acceptable salt
thereof for the inhibition of platelet aggregation.
[0032] The invention is furthermore directed to a pharmaceutical
composition comprising a pharmacologically active agent and a
modified amino acid, or pharmaceutically acceptable salt thereof,
wherein the modified amino acid or salt thereof is present in an
amount effective to inhibit platelet aggregation for the inhibition
of platelet aggregation.
[0033] Further features and advantages of the invention will become
apparent from the following detailed description of the invention
and the appended claims.
[0034] A dose-inhibition experiment in platelet rich plasma (PRP)
from 12 healthy subjects using 5 .mu.M adenosine diphosphate (ADP)
as platelet aggregation stimulator and various concentrations of
5-CNAC as platelet aggregation inhibitor was made. The platelet
aggregation curves for three of the individual subjects stimulated
by 5 .mu.M ADP were established.
[0035] A dose-inhibition experiment was made in platelet rich
plasma (PRP) from 12 healthy subjects using 5 .mu.M adenosine
diphosphate (ADP) as platelet aggregation stimulator and various
concentrations of 5-CNAC as platelet aggregation inhibitor.
Platelet aggregation curves for three of the individual subjects
stimulated by 5 .mu.M ADP were established.
[0036] A dose-inhibition experiment was made in PRP using 3 .mu.M
ADP as platelet aggregation stimulator and various concentrations
of 5-CNAC as platelet aggregation inhibitor. The platelet
aggregation curves for two of the individual subjects stimulated by
3 .mu.M ADP were established.
[0037] A dose-inhibition experiment in PRP using 2 .mu.M ADP as
platelet aggregation stimulator and various concentrations of
5-CNAC as platelet aggregation inhibitor was made. The platelet
aggregation curves for four of the individual subjects stimulated
by 2 .mu.M ADP were established.
[0038] A dose-inhibition experiment in PRP using 5 .mu.g/mL
collagen as platelet aggregation stimulator and various
concentrations of 5-CNAC as platelet aggregation inhibitor was
made. The platelet aggregation curves for two of the individual
subjects stimulated by 5 .mu.g/mL collagen were established.
[0039] A dose-inhibition experiment in PRP using 2.5 .mu.g/mL
collagen as platelet aggregation stimulator and various
concentrations of 5-CNAC as a platelet aggregation inhibitor was
made. The platelet aggregation curves for two of the individual
subjects stimulated by 2.5 .mu.g/mL collagen were established.
[0040] A dose-inhibition experiment in PRP using 2.0 .mu.g/mL
collagen as platelet aggregation stimulator and various
concentrations of 5-CNAC as a platelet aggregation inhibitor was
made. The platelet aggregation curve for the subject stimulated by
2.0 .mu.g/mL collagen were established.
[0041] A dose-inhibition experiment in PRP using 1 .mu.g/mL
collagen as platelet aggregation stimulator and various
concentrations of 5-CNAC as a platelet aggregation inhibitor was
made. The platelet aggregation curves for the subjects stimulated
by 1 .mu.g/mL collagen were established.
[0042] A dose-inhibition experiment in PRP using 0.75 .mu.g/mL
collagen as platelet aggregation stimulator and various
concentrations of 5-CNAC as a platelet aggregation inhibitor was
made. A dose-inhibition experiment in PRP using 0.5 .mu.g/mL
collagen as platelet aggregation stimulator and various
concentrations of 5-CNAC as a platelet aggregation inhibitor
inhibitor was made. The platelet aggregation curves for the
subjects stimulated by 0.5 .mu.g/mL collagen were established.
[0043] The modified amino acids useful in the present invention
include any one of the 123 modified amino acids disclosed in
aforementioned '536 or any one of the 193 modified amino acids
described in the aforementioned '647 or any combination thereof.
The contents of the aforementioned '647 and '536 are hereby
incorporated by reference in their entirety, especially the subject
matter of the claims and corresponding working examples. In
addition, the modified amino acids can be the disodium salt of any
of the aforementioned modified amino acids as well as ethanol
solvates and hydrates thereof. Suitable compounds include compounds
of the following formula I ##STR2## wherein [0044] R.sup.1,
R.sup.2, R.sup.3 and R.sup.4 are independently hydrogen, --OH,
--NR.sup.6R.sup.7, halogen, C.sub.1-C.sub.4alkyl or
C.sub.1-C.sub.4alkoxy; [0045] R.sup.5 is a substituted or
unsubstituted C.sub.2-C.sub.16alkylene, substituted or
unsubstituted C.sub.2-C.sub.16alkenylene, substituted or
unsubstituted C.sub.1-C.sub.12alkyl(arylene), or substituted or
unsubstituted aryl(C.sub.1-C.sub.12alkylene); and [0046] R.sup.6
and R.sup.7 are independently hydrogen, oxygen or
C.sub.1-C.sub.4alkyl; and hydrates and alcohol solvates thereof.
The compounds of formula I as well as their disodium salts and
alcohol solvates and hydrates thereof are described in WO
00/059863, along with methods for preparing them.
[0047] The disodium salt may be prepared from the ethanol solvate
by evaporating or drying the ethanol solvate by methods known in
the art to form the anhydrous disodium salt. Drying is generally
carried out at a temperature of from about 80.degree. C. to about
120.degree. C., preferably from about 85.degree. C. to about
90.degree. C., and most preferably at about 85.degree. C. The
drying step is generally performed at a pressure of 26'' Hg or
greater. The anhydrous disodium salt generally contains less than
about 5% by weight of ethanol and preferably less than about 2% by
weight of ethanol, based on 100% total weight of anhydrous disodium
salt.
[0048] The disodium salt of the modified amino acid can also be
prepared by making a slurry of the modified amino acid in water and
adding two molar equivalents of aqueous sodium hydroxide, sodium
alkoxide or the like. Suitable sodium alkoxides include, but are
not limited to, sodium methoxide, sodium ethoxide and combinations
thereof.
[0049] A still further method of preparing the disodium salt is by
reacting the modified amino acid with one molar equivalent of
sodium hydroxide to yield the disodium salt.
[0050] The disodium salt can be isolated as a solid by
concentrating the solution containing the disodium salt to a thick
paste by vacuum distillation. This paste may be dried in a vacuum
oven to obtain the disodium salt of the modified amino acid as a
solid. The solid can also be isolated by spray drying an aqueous
solution of the disodium salt.
[0051] The modified amino acids may be prepared by methods known in
the art, e.g., as mentioned above, by methods described in '647 and
'536.
[0052] The ethanol solvates, as described in the aforementioned
'863, include, but are not limited to, a molecular or ionic complex
of molecules or ions of ethanol solvent with molecules or ions of
the disodium salt of the modified amino acid. Typically, the
ethanol solvate contains about one ethanol molecule or ion for
every molecule of disodium salt of the modified amino acid.
[0053] The ethanol solvate of the disodium salt of the modified
amino acid can be prepared by dissolving the modified amino acid in
ethanol. Typically, each gram of modified amino acid Is dissolved
in from about 1 mL to about 50 mL of ethanol and generally, from
about 2 mL to about 10 mL of ethanol. The modified amino
acid/ethanol solution is then reacted with a molar excess of a
sodium containing salt, such as a monosodium containing salt,
relative to modified amino acid, i.e., for every mole of modified
amino acid there is more than one mole of sodium cations, yielding
the ethanol solvate. Suitable monosodium salts include, but are not
limited to, sodium hydroxide; sodium alkoxides, such as sodium
methoxide and sodium ethoxide; and any combination of the
foregoing. Preferably, at least about two molar equivalents of the
monosodium containing salt are added to the ethanol solution, i.e.,
for every mole of modified amino acid there is at least about two
moles of sodium cations. Generally, the reaction is performed at or
below the reflux temperature of the mixture, such as at ambient
temperature. The ethanol solvate is then recovered by methods known
is the art, such as concentration of the resulting slurry at
atmospheric distillation, cooling the concentrated slurry and
filtering the solid. The recovered solid can then be vacuum dried
to obtain the ethanol solvate.
[0054] The hydrates of the disodium salts of the modified amino
acids may be prepared by drying the ethanol solvate to from an
anhydrous disodium salt, as described above, and hydrating the
anhydrous disodium salt. Preferably, the monohydrate of the
disodium salt is formed. Since the anhydrous disodium salt is very
hydroscopic, the hydrate forms upon exposure to atmospheric
moisture. Generally, the hydrating step is performed at from about
ambient temperature to about 50.degree. C., preferably ambient
temperature to about 30.degree. C. and in an environment having at
least 50% relative humidity. Alternatively, the anhydrous disodium
salt may be hydrated with steam.
[0055] The preferred modified amino acids are 5-CNAC, also known as
8-(N-2-hydroxy-5-chlorobenzoyl)aminocaprylic acid, SNAD, SNAC and
their monosodium and disodium salts, ethanol solvates of their
sodium salts and the monohydrates of their sodium salts and any
combinations thereof. The most preferred modified amino acid is the
disodium salt of 5-CNAC and the monohydrate thereof.
[0056] The pharmacologically active agents suitable for use in the
instant invention include both therapeutic as well as preventative
agents. The pharmacologically active agents include, but are not
limited to proteins, polypeptides, hormones, polysaccharides
including mixtures of muco-polysaccharides, carbohydrates, lipids
and combinations thereof.
[0057] Specific examples of pharmacologically active agents
include, but are not limited to the following, including synthetic,
natural or recombinant sources thereof: growth hormone, including
human growth hormones, recombinant human growth hormones, bovine
growth hormones and porcine growth hormones; growth
hormone-releasing hormones; interferons, including .alpha., .beta.
and .gamma.-interferon; interleukin-1; interleukin-2; insulin,
including porcine, bovine, human and human recombinant, optionally
having counter ions including sodium, zinc, calcium and ammonium;
insulin-like growth factor, including IGF-1; heparin, including
unfractionated heparin, heparinoids, dermatans, chondroitins, low,
very low and ultra low molecular weight heparins; calcitonin,
including salmon, porcine, eel, chicken and human; erythropoietein;
atrial naturetic factor; antigens; monoclonal antibodies;
somatostatin; protease inhibitors; adrenocorticotropin,
gonadotropin releasing hormone; oxytocin;
leutinizing-hormone-releasing hormone; follicle stimulating
hormone; glucocerebrosidase; thrombopoietin; filgrastim;
prostaglandins; cyclosporin; vasopressin; cromolyn sodium (sodium
or disodium chromoglycate); vancomycin; desferrioxamine;
parathyroid hormone, including its fragments; antimicrobials,
including anti-fungal agents; vitamins; analogs, fragments,
mimetics or polyethylene glycol-modified derivatives of these
compounds or any combination thereof.
[0058] A preferred pharmacologically active agent is a
pharmacologically active peptide, particularly calcitonin. A known
class of pharmacologically active agents, calcitonins have varying
pharmaceutical utility and are commonly employed in the treatment
of, e.g., Paget's disease, hypercalcemia and postmenopausal
osteoporosis. Various calcitonins, including salmon, pig and eel
calcitonin are commercially available and commonly employed for the
treatment of, e.g., Paget's disease, hypercalcemia of malignancy
and osteoporosis. The calcitonin can be any calcitonin, including
natural, synthetic or recombinant sources thereof, as well as
calcitonin derivatives, such as 1,7-Asu-eel calcitonin. The
compositions can comprise a single calcitonin or any combination of
two or more calcitonins. The preferred calcitonin is synthetic
salmon calcitonin. The calcitonins are commercially available or
may be synthesized by known methods.
[0059] Other preferred pharmacologically active agents are heparin,
insulin and PTH.
[0060] The amount of pharmacologically active agent is generally an
amount effective to accomplish the intended purpose, e.g., a
therapeutically effective amount. However, the amount can be less
than that amount when a plurality of the compositions are to be
administered, i.e., the total effective amount can be administered
in cumulative dosage units. The amount of active agent can also be
more than the effective amount when the composition provides
sustained release of the pharmacologically active agent. The total
amount of active agent to be used can be determined by methods
known to those skilled in the art. However, because the
compositions may deliver the active agent more efficiently than
prior compositions, less amounts of active agent than those used in
prior dosage unit forms or delivery systems can be administered to
a subject while still achieving the same blood levels and/or
therapeutic effects.
[0061] When the pharmacologically active agent is salmon
calcitonin, the appropriate dosage will, of course, vary depending
upon, for example, the host and the nature and severity of the
condition being treated. However, in general, satisfactory results
will be obtained systemically at daily dosages of from about 0.5
.mu.g/kg to about 10 .mu.g/kg animal body weight, preferably 1
.mu.g/kg to about 6 .mu.g/kg body weight.
[0062] The pharmacologically active agent generally comprises from
0.05% to 70% by weight relative to the total weight of the overall
pharmaceutical composition, preferably an amount of from 0.01% to
50% by weight, more preferably 0.3% to 30% by weight relative to
the total weight of the overall pharmaceutical composition.
[0063] The pharmaceutical compositions for use in the present
invention typically comprises a platelet-aggregation-inhibitory
amount of one or more of the modified amino acids, i.e., an amount
sufficient to inhibit blood platelet aggregation. Generally, the
modified amino acid is present in a dosage range of between about
25 mg and about 400 mg. Most preferably the modified amino acid is
present in a dosage range of between about 100 mg and about 200
mg.
[0064] The pharmaceutical compositions for use in the present
invention typically comprises a pharmaceutically active agent and a
platelet-aggregation-inhibitory amount of one or more of the
modified amino acids, i.e., an amount sufficient to inhibit blood
platelet aggregation.
[0065] The pharmaceutical compositions for use in the present
invention may be provided as a capsule including a soft-gel
capsule, tablet, caplet or other solid oral dosage form, all of
which can be prepared by methods well-known in the art.
[0066] The pharmaceutical compositions for use in the present
invention may additionally comprise additives in amounts
customarily employed including, but not limited to, a pH adjuster;
a preservative; a flavorant; a taste-masking agent; a fragrance; a
humectant; a tonicifier; a colorant; a surfactant; a plasticizer; a
lubricant, such as magnesium stearate; a flow aid; a compression
aid; a solubilizer; an excipient; a diluent, such as
microcrystalline cellulose, e.g., Avicel PH 102 supplied by FMC
corporation; or any combination thereof. Other additives may
include phosphate buffer salts, citric acid, glycols and other
dispersing agents.
[0067] The pharmaceutical compositions for use in the present
invention may optionally additionally comprise crospovidone, which
can be any crospovidone. Crospovidone is a synthetic crosslinked
homopolymer of N-vinyl-2-pyrrolidone, also called
1-ethenyl-2-pyrrolidinone, having a molecular weight of 1,000,000
or more. Commercially available crospovidones include Polyplasdone
XL, Polyplasdone XL-10, Polyplasdone INF-10 available from ISP,
Kollidon CL, available from BASF Corporation.
[0068] Povidone is a synthetic polymer consisting of linear
1-vinyl-2-pyrrolidinone groups having a molecular weight generally
between 2,500 and 3,000,000. Commercially available povidones
include Kollidon K-30, Kollidon K-90F available from BASF
Corporation and Plasdone K-30 and Plasdone K-29/32, available from
ISP.
[0069] As mentioned above, the crospovidones and povidones are
commercially available. Alternatively, they may be synthesized by
known processes.
[0070] The crospovidone, povidone or combination thereof is
generally present in the compositions in an amount of from 0.5% to
50% by weight relative to the total weight of the overall
pharmaceutical composition, preferably an amount of from 2% to 25%,
more preferably 5% to 20% by weight relative to the total weight of
the pharmaceutical composition.
[0071] The pharmaceutical composition may also include one or more
enzyme inhibitors, such as actinonin or epiactinonin and
derivatives thereof; aprotinin, Trasylol and Bowman-Birk
inhibitor.
[0072] Further, a transport inhibitor, i.e., a .rho.-glycoprotein,
such as Ketoprofin, may be present in the compositions of the
present invention.
[0073] Preferably, the solid pharmaceutical compositions of the
instant invention include a diluent, such as Avicel; and a
lubricant, such as magnesium stearate.
[0074] The solid pharmaceutical compositions of the instant
invention can be prepared by conventional methods, e.g., by
blending a mixture of the active agent or active agents, the
delivery agent and other ingredients, kneading and filling into
capsules, or instead of filling into capsules, molding followed by
further tableting or compression-molding to give tablets. In
addition, a solid dispersion may be formed by known methods
followed by further processing to form a tablet or capsule.
[0075] Preferably, the ingredients in the pharmaceutical
compositions of the instant invention are homogeneously or
uniformly mixed throughout the solid dosage form.
[0076] The pharmaceutical compositions of the present invention may
be administered to deliver a pharmacologically active agent to any
mammal in need thereof including, but not limited to, rodents,
cows, pigs, dogs, cats and primates, particularly humans.
EXPERIMENTAL PROCEDURES
[0077] Preparation of PRP (Platelet Rich Plasma)
[0078] Freshly drawn venous blood from healthy volunteers is
collected into 0.1 vol. mmol/L trisodium citrate. The donors have
not taken any medication during two weeks prior to blood
collection. PRP is prepared by centrifugation of the freshly drawn
blood (150 g, 15 minutes at 22.degree. C.) and the final platelet
concentration is standardized at 200 000 cell/.mu.L by dilution in
autologous platelet-free plasma, prepared by centrifugation (1200
g, 10 minutes at 22.degree. C.). The supernatant is collected.
Samples of PRP are pre-incubated for 1 minute (22.degree. C.) with
varying concentrations of 5-CNAC in saline (stored at -20.degree.
C.).
[0079] Platelet Aggregation Studies
[0080] In order to assess the inhibitory effect of 5-CNAC on
platelet aggregation, the following studies are performed. Samples
of 0.400 mL PRP containing 5-CNAC are prepared as above. To these
samples is added 0.005 mL of an aggregation stimulator which is
either ADP (Roche Molecular Biochemicals, Mannheim) or collagen.
(Horm Chemie, Munich). The final concentrations of either the ADP
or collagen are as indicated in the "Results" section. Aggregation
studies are performed in a Chronolog 4 channel aggregometer (type
CH570VS-CH810). The aggregometer automatically calibrates the
donor-dependent difference in optical density between platelet-free
plasma (0% optical density) and the PRP (100% optical density) of a
particular donor. The extent of aggregation is measured by
expressing the maximal difference in optical density (independent
of the time after the addition of the aggregation stimulator, i.e.,
either ADP or collagen) and normalized taking the control curves
(without 5-CNAC) as an internal standard set at 100%.
[0081] Results
[0082] Effect of 5-CNAC on Platelet Aggregation Induced by ADP
[0083] It is noted that the addition of 5-CNAC alone to PRP does
not trigger platelet aggregation.
[0084] A dose-inhibition experiment was made in PRP from 12 healthy
subjects using 5 .mu.M ADP as the platelet aggregation stimulating
agent and varying concentrations of 5-CNAC as the platelet
aggregation inhibiting agent. Concentrations tested include 0.1, 1,
2, 5, 10, 25, 100, 200 and 500 .mu.M It is seen that at increasing
concentrations of 5-CNAC, inhibition of platelet aggregation
becomes apparent at about 2 .mu.M 5-CNAC or more. The aggregation
curves of three individual subjects induced by 5.mu.M ADP were
established. It is noted that 5 .mu.M ADP stimulates maximal
platelet aggregation.
[0085] In order to study the inhibitory effect of 5-CNAC at
suboptimal ADP stimulation, studies were repeated with lower
concentrations of ADP challenged by varying concentrations of
5-CNAC. A dose-inhibition study was made using 3 .mu.M ADP. The
corresponding aggregation curves for individual subjects were
established . The same experiments using 2 .mu.M ADP as the
stimulator was made. The aggregation curves of individual subjects
were established. The result showed that, as the concentration of
the platelet aggregation stimulator (ADP) decreases, platelet
aggregation is inhibited by lower concentrations of 5-CNAC.
Moreover, it is observed that in all concentrations of ADP tested,
the platelet aggregation inhibition by 5-CNAC is evident in the
second phase of the aggregation curve, but is not observed in the
initial phase of the aggregation curve. This effect is similar to
integrin IIb 3 (glycoprotein IIb-IIIa) antagonists that interfere
with platelet-platelet coupling.
[0086] Effect of 5-CNAC on Platelet Aggregation Stimulated by
Collagen
[0087] Compared with ADP, collagen is a more potent platelet
aggregation inducer. There is an adhesion phase in which platelets
bind to the collagen insoluble fibers before becoming activated.
The result shows that platelets are relatively resistant to
aggregation inhibition when the platelets are stimulated by a high
concentration of the aggregation stimulator collagen (5 .mu.g/mL).
Platelets are unaffected by 100 .mu.M 5-CNAC. However, at about 1
mM 5-CNAC, inhibition of platelet aggregation becomes
detectable.
[0088] Experiments were made at lower collagen concentrations (2.5
.mu.g/mL) and showed that there is some inhibition of platelet
aggregation by 200 .mu.M 5-CNAC in the platelet rich plasma of one
individual, but not at lower concentrations of the 5-CNAC At 2
.mu.g/mL collagen, there is one individual showing platelet
aggregation inhibition at about 1 mM 5-CNAC, while at a lower
collagen concentration (1 .mu.g/mL) inhibition by 5-CNAC is evident
at 50 .mu.M or more. Lower concentrations of the stimulator
collagen reveal similar inhibition patterns by 5-CNAC. As with
ADP-stimulated aggregation, the inhibitory effect of 5-CNAC on
collagen-stimulated aggregation becomes apparent in the second
phase of the aggregation curve, while no aggregation inhibition is
observed in the primary aggregation curve. Also, when the
concentration of collagen is lowered, the platelets become more
sensitive to 5-CNAC inhibition.
[0089] Further observations include the following: i) 5-CNAC does
not alter platelet morphology since the "swirling" of the cells
remains intact; (ii) upon ADP addition there is an increase in
optical density. This downward pattern is caused by a change in
cell shape, which is a first response to platelet aggregation. This
response is undisturbed in the presence of 5-CNAC; (iii) 5-CNAC
dose-dependently inhibits the later part of the aggregation curve
(also called secondary aggregation); (iv) the sensitivity of the
PRP to 5-CNAC differs between subjects; and (v) the effect of
5-CNAC shows similarities with the inhibition by aspirin-like drugs
that interfere with thromboxane A.sub.2 production, or platelets
from patients with a congenital secretion defect (so-called
"storage pool deficiency").
[0090] As can be seen from the foregoing, modified amino acids of
the instant invention are effective at inhibiting platelet
aggregation.
* * * * *