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Description  |
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BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to the use of tenidap and the
pharmaceutically-acceptable base salts thereof for inhibiting the release
of elastase by neutrophils in a mammal. Tenidap and its salts are useful
for inhibiting the release of elastase by neutrophils in a mammal, per se,
and in treating elastase-mediated diseases and dysfunctions in a mammal.
Such elastase-mediated diseases and dysfunctions include, but are not
limited to, arteritis, proteinuria and pulmonary emphysema. The use of
tenidap and its salts comprises administering an effective amount thereof
to a mammal.
2. General Background
Tenidap,
5-chloro-2,3-dihydro-2-oxo-3-(2-thienylcarbonyl)-indole-1-carboxamide, has
the structural formula
##STR1##
Tenidap, among other 3-substituted-2-oxindole-1-carboxamides are disclosed
and claimed in U.S. Pat. No. 4,556,672 which is assigned to the assignee
hereof. That patent discloses that those compounds, in addition to being
useful as antiinflammatory and analgesic agents, are inhibitors of both
cyclooxygenase (CO) and lipoxygenase (LO). The teachings thereof are
incorporated herein by reference.
The use of tenidap and its pharmaceutically-accceptable base salts, among
certain other 3-substituted-2-oxindole-1-carboxamides, to inhibit
interleukin-1 biosynthesis in a mammal and to treat interleukin-1 mediated
disorders and dysfunctions is disclosed in U.S. Pat. No. 4,861,794 which
is assigned to the assignee hereof.
U.S. Pat. No. 4,853,409, assigned to the assignee hereof, discloses the use
of tenidap and its pharmaceutically-acceptable base salts, among certain
other 3-substituted-2-oxindole-1-carboxamides, to suppress T-cell function
in a mammal and to treat T-cell mediated autoimmune disorders of the
systemic or organ specific type.
An anhydrous, crystalline form of the sodium salt of tenidap is disclosed
in European Patent Application 277,738, which has been filed in the name
of the assignee hereof.
Elastase is a protease which is released by neutrophils in a mammal and
mediates certain diseases and dysfunctions. [Janoff, A., American Journal
of Pathology 68:579-591 (1972).] Such elastase mediated diseases and
dysfunctions include, but are not limited to, arteritis, proteinuria and
pulmonary emphysema [Janoff, A., Op. cit. and Johnson, R. J., et al., J.
Exp. Med. 168:1169-1174 (1988).]
Until the invention herein, there was no report of use or intent to use
tenidap or its salts to inhibit release of elastase by neutrophils in a
mammal and to treat elastase-mediated diseases and dysfunctions with such
compounds nor any appreciation of their role in such treatments.
SUMMARY OF THE INVENTION
It has been found that tenidap and the pharmaceutically-acceptable base
salts thereof inhibit the release of elastase by neutrophils and are
useful in inhibiting the release of elastase by neutrophils in a mammal,
per se, and in treating elastase-mediated diseases and dysfunctions. Such
elastase-mediated diseases and dysfunctions include, but are not limited
to, arteritis, proteinuria and pulmonary emphysema.
The methods of using tenidap and its pharmaceutically-acceptable base salts
comprise administering to a mammal an effective amount thereof.
Administration can comprise any known method for therapeutically providing
a compound to a mammal such as by oral or parenteral administration as
defined hereinbelow.
DETAILED DESCRIPTION OF THE INVENTION
Tenidap, which has the chemical structure
##STR2##
its pharmaceutically-acceptable base salts and the preparation thereof
are described in U.S. Pat. No. 4,556,672, the teachings of which are
incorporated herein by reference. This invention concerns new uses for
tenidap and its salts which comprise methods for inhibiting the release of
elastase by neutrophils in a mammal in need thereof. Also within the scope
of this invention are methods of treating elastase-mediated disorders and
dysfunctions in a mammal which include, but are not limited to, arteritis,
proteinuria and pulmonary emphysema.
As disclosed in U.S. Pat. No. 4,556,672, tenidap is acidic and forms base
salts. All such base salts are within the scope of this invention and can
be formed as taught by that patent. Such suitable salts, within the scope
of this invention, include both the organic and inorganic types and
include, but are not limited to, the salts formed with ammonia, organic
amines, alkali metal hydroxides, alkali metal carbonates, alkali metal
bicarbonates, alkali metal hydrides, alkali metal alkoxides, alkaline
earth metal hydroxides, alkaline earth metal carbonates, alkaline earth
metal hydrides and alkaline earth metal alkoxides. Representative examples
of bases which form such base salts include ammonia, primary amines, such
as n-propylamine, n-butylamine, aniline, cyclohexylamine, benzylamine,
p-toluidine, ethanolamine and glucamine; secondary amines, such as
diethylamine, diethanolamine, N-methylglucamine, N-methylaniline,
morpholine, pyrrolidine and piperidine; tertiary amines, such as
triethylamine, triethanolamine, N,N-dimethylaniline, N-ethylpiperidine and
N-methylmorpholine; hydroxides, such as sodium hydroxide; alkoxides such
as sodium ethoxide and potassium methoxide; hydrides such as calcium
hydride and sodium hydride; and carbonates such as potassium carbonate and
sodium carbonate. Preferred salts are those of sodium, potassium,
ammonium, ethanolamine, diethanolamine and triethanolamine. Particularly
preferred is the sodium salt. European Patent Application 277,738, which
has been filed in the name of the assignee hereof, discloses an anhydrous,
crystalline form of such a salt. The teachings thereof are incorporated
herein by reference.
Also within the scope of this invention are the solvates such as the
hemihydrates and monohydrates of the compounds hereinabove described.
The methods of this invention comprise administering tenidap and the
pharmaceutically-acceptable base salts thereof to a mammal. Such compounds
and their salts can be administered to said mammal either alone or,
preferably, in combination with pharmaceutically-acceptable carriers or
diluents in a pharmaceutical composition, according to standard
pharmaceutical practice. Such administration can be oral or parenteral.
Parenteral administration as used herein includes, but is not limited to,
intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal and
topical including, but not limited to oral lavage and inhalation,
administration. While it is generally preferred to administer such
compounds and their salts orally, other methods may be preferred depending
upon the particular elastase-mediated disease or dysfunction being
treated.
In general, tenidap and its salts are most desirably administered in doses
ranging from about 20 mg up to about 200 mg per day, with a preferred
range of about 40 mg up to about 120 mg per day, for oral administration
and from about 1 mg up to about 200 mg per day for parenteral
administration, although variations will still necessarily occur depending
upon the weight of the subject being treated. The appropriate dose for
inhibiting the release of elastase by neutrophils in a mammal and for
treatment of elastase-mediated disorders and dysfunctions with tenidap and
its salts will be readily determined by those skilled in the art of
prescribing and/or administering such compounds. Nevertheless, it is still
to be appreciated that other variations may also occur in this respect,
depending upon the species of mammal being treated and its individual
response to said medicament, as well as on the particular type of
pharmaceutical formulation chosen and the time period and interval at
which such administration is carried out. In some instances, dosage levels
below the lower limit of the aforesaid range may be more than adequate,
while in other cases still larger doses may be employed without causing
any harmful or deleterious side effects to occur, provided that such
higher dose levels are first divided into several smaller doses that are
to be administered throughout the day.
For purposes of oral administration, tablets containing excipients such as
sodium citrate, calcium carbonate and dicalcium phosphate may be employed
along with various disintegrants such as starch and preferably potato or
tapioca starch, alginic acid and certain complex silicates, together with
binding agents such as polyvinylpyrrolidone, sucrose, gelatin and acacia.
Additionally, lubricating agents such as, but not limited to, magnesium
stearate, sodium lauryl sulfate and talc are often very useful for
tableting purposes. Solid compositions of a similar type may also be
employed as fillers in soft elastic and hard-filled gelatin capsules;
preferred materials in this connection also include, by way of example and
not of limitation, lactose or milk sugar as well as high molecular weight
polyethylene glycols. When aqueous suspensions and/or elixirs are desired
for oral administration, the essential active ingredient may be combined
with various sweetening or flavoring agents, coloring matter or dyes and,
if so desired, emulsifying and/or suspending agents, together with
diluents such as water, ethanol, propylene glycol, glycerin and various
like combinations thereof.
Although the generally preferred mode of administration of tenidap or its
pharmaceutically-acceptable base salts is oral, they may be administered
parenterally as well. Such parenteral administration may be the preferred
mode of administration for the treatment of certain elastase-mediated
diseases or dysfunctions.
For purposes of parenteral administration, solutions of tenidap or a salt
thereof in sesame or peanut oil or in aqueous propylene glycol may be
employed, as well as sterile aqueous solutions of the corresponding water
soluble base salts previously enumerated. Such aqueous solutions should be
suitably buffered if necessary, and the liquid diluent rendered isotonic
with sufficient saline or glucose. These particular aqueous solutions are
especially suitable for intravenous, intramuscular and subcutaneous
injection purposes. In this connection, the sterile aqueous media employed
are readily obtained by standard techniques well known to those skilled in
the art. For instance, distilled water is ordinarily used as the liquid
diluent and the final preparation is passed through a suitable bacterial
filter such as a sintered glass filter or a diatomaceous-earth or unglazed
porcelain filter. Preferred filters of this type include the Berkefeld,
the Chamberland and the Asbestos Disk-Metal Seitz filter, wherein the
fluid is sucked into a sterile container with the aid of a suction pump.
The necessary steps should be taken throughout the preparation of these
injectable solutions to insure that the final products are obtained in a
sterile condition. For purposes of transdermal administration, the dosage
form of the particular compound may include, by way of example, solutions,
lotions, ointments, creams, gels, suppositories, rate-limiting sustained
release formulations and devices therefor. Such dosage forms comprise the
particular compound and may include ethanol, water, penetration enhancer
and inert carriers such as gel-producing materials, mineral oil,
emulsifying agents, benzyl alcohol and the like. Specific transdermal flux
enhancing compositions are disclosed in European Patent Application
271,983 and European Patent Application 331,382, which have been filed in
the name of the assignee of this invention, the teachings of which are
incorporated herein by reference. For purposes of topical administration,
the dosage form of the particular compound may include, by way of example
and not of limitation, solutions, lotions, ointments, creams and gels.
Further, administration by inhalation can be achieved by menas and methods
well known to those skilled in the art. Such means include the use of
nebulizers or atomizers whereby a solution of tenidap or a salt thereof is
inhaled as a mist.
The ability of the compounds of this invention to inhibit the release of
elastase by neutrophils was demonstrated by the assay procedure described
in Dunn, T. L., et al., Analytical Biochemistry 150:18-24 (1985) and
references cited therein. Neutrophils for the assay were obtain as
follows. Whole human blood from normal volunteers was obtained by
venipuncture into heparinized syringes. The majority of the red cells were
removed by dextran sedimentation and neutrophils were separated by density
centrifugation over hypaque ficoll. The neutrophil rich fraction was
washed and residual red cells were removed by hypotonic lysis according to
the procedure described by Blackburn, W. D. et al., Arthritis Rheum.
30:1006-1014 (1987). The neutrophils so prepared were used in the assay
described below and cell viability was assured by determining their
ability to exclude typan blue. In each assay the cell viability routinely
exceeded 95%.
Affinity purified anti-human neutrophil elastase (anti-HNE) antibody was
labeled with carrier free .sup.125 I-Na by using a modification of the
lactoperoxidase method of Marchelonis, J. J., Biochem. J. 113:229-305
(1969). Generally, 10 .mu.g quantities of protein were labeled to an
initial specific activity of 2.2.times.10.sup.-5 mCi/ng. Free iodine was
separated from bound .sup.125 I by Sephadex G-25 column chromatography.
The .sup.125 I-labeled anti-HNE was aliquoted and stored at -70.degree. C.
for up to one month prior to use.
Neutrophil cell suspensions, prepared as described above, were incubated at
37.degree. C. for 15-30 minutes in the presence of varying concentrations
of tenidap. Tenidap was dissolved and diluted in water and added to the
cells directly therefrom. After the cells had been incubated in the
presence of tenidap, the cell suspensions (5.times.10.sup.6 cells/ml, 125
.mu.l well) were added to IgG coated and bovine serum albumin (BSA)
blocked wells of microtiter plates and incubated for 45 minutes at
37.degree. C. As controls, similar incubations were performed in the
absence of IgG. Following incubation, the cell suspensions were
centrifuged (750.times.g) for 5 minutes at 4.degree. C.
DE-52 cellulose-purified IgG fraction goat anti-HNE (10 mg/ml), diluted at
1/1000 in PBS, was used to coat vinyl assay wells (125 .mu.l ) for 4 hours
at 25.degree. C. The wells were then blocked with PBS-1% BSA (100 .mu.l )
for 1 hour at 25.degree. C. to eliminate nonspecific binding, washed with
PBS three times and 100 .mu.l samples of the supernatants obtained as
described above were then added to each well and allowed to incubate 16
hours at 25.degree. C. Standard curves were generated with serial
dilutions of the DFP inactivated enzyme (500 .mu.g/ml) in PBS-1% BSA.
After three washings with PBS, affinity purified .sup.125 I-labeled
anti-HNE was added to each well (100,000 cpm/100 .mu.l ). The wells were
incubated for 16 hours at 25.degree. C. and washed three times with PBS
and each well was counted for 1 minute in a gamma counter. .sup.125
I-Anti-HNE (cpm bound).times.10.sup.-3 was plotted against protein
concentration in nanograms per milliliter. Standard binding curves using
other purified proteins instead of HNE were used as described above.
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