Note: Descriptions are shown in the official language in which they were submitted.
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USE OF PDE INHIBITORS IN THE MANUFACTURE OF A MEDICAMENT FOR THE TREATMENT
OF BLADDER DISEASES
The present invention pertains to the use of inhibitors of
phosphodiesterase I (PDE I) for the treatment of urinary bladder
diseases.
The physiological transmission of information for the relaxation
of smooth muscle cells is effected by transmitters (messengers)
of the blood (hormones) or the nerves (neurotransmitters). These
messengers and neurotransmitters cause an increase in the levels
of the cyclic nucleotides "cyclic adenosine monophosphate" (cAMP)
and "cyclic guanosine monophosphate" (cGMP) in the smooth muscle
cell, resulting in relaxation. cAMP and cGMP themselves are
hydrolyzed by phosphodiesterases (PDEs). Inhibitors of the PDEs
in turn reduce the hydrolyzation of cAMP and cGMP, resulting in
an lncrease of these molecules within the cell and thus in a
relaxation of the smooth muscle cell. This mechanism of actlon
has been described, for instance, by C.D. Nicholson, R.A.
Challiss, and M. Shadid: Pulm. Pharmacol. 7 (1) (1994), 1-17, and
T.J. Torphy et al.: J. Pharmacol. Exp. Ther. 265 (3) (1993), 1213-
23.
From these publications as well as from W.J. Thompson: Pharmacol.
Ther. 51 (1991), 13-33, and J. Beavo in: J. Beavo and M.D. Housley
(eds.): Cyclic nucleotide phosphodiesterases: Structure, regula-
tion and drug action, Chichester, New YorkBrisbane Toronto-Singa-
pore, Wiley, 1990: 3-15; there is further known the distinction
of a number of isoenzymes of PDEs is distinguished between five
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different PDEs (I - V) which are differently distributed in the
individual organs and organ systems and have different activities
according to their distribution. In the publications mentioned,
there is also discussed the occurrence of the different isoenzymes
in various tissues.
An interesting target for the use of PDE isoenzyme selective
inhibitors is the lower urinary tract since the pharmacological
therapy of bladder dysfunctions with conventional substances is
often little effective and limited by side-effects. Therefore,
a well-aimed effect on the bladder muscle by inhibiting a
functionally important PDE isoenzyme appears to be superior to
established pharmacological treatment modalities.
According to the invention it has surprisingly been found that
inhibitors of phosphodiesterase I in the preparation of a drug
can be used in the treatment of urinary bladder diseases. Pre-
ferred embodiments of the use according to the invention are
related with claims 1 - 4. The claim 5 deals with a method for
treating urinary bladder diseases by administering an inhibitor
of PDE I in a sufficient amount to a patient in need thereof. It
is preferred that the formulation is given in a galenically
acceptable formulation.
According to the invention a method of identifying and classifying
inhibitors of PDE I is disclosed by competitive studies using
inhibitors of PDE I as competing agents.
Surprisingly, it has now been found that PDE I is of particular
importance in human bladder muscle: A well-aimed inhibition o~
this isoenzyme will result in relaxation of the bladder muscle
even if minute doses of an inhibitor are administered, e.g., the
PDE I inhibitor vinpocetine in a dosage o~ 10 mol/l (figure 2),
with no appreciable effects to other organs, in particular to
vessels. Therefore, there is an excellent efficacy and potency
in the treatment of urinary bladder diseases.
-
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Therefore, the subject matter of the invention is the use of PDE
I inhibitors in the treatment of urinary bladder diseases, in
particular the so-called urge symptoms, frequency, urge incon-
tinence, involuntary discharge of urine, and instabilities of the
bladder muscle (detrusor instability), and the use of the inhibi-
tors for the preparation of drugs useful for this purpose as well
as drugs containing PDE I inhibitors for the objects mentioned.
Preferred inhibitors of PDE I are:
vincamine, vinpocetine (ethyl apovincamin-22-oate),
and/or modified compounds of the said inhibitors having inhibiting
properties for PDE I,
as well as pharmacologically compatible salts thereof.
The pharmacologically compatible salts are obtained in a similar
manner by neutralizing the bases with inorganic or organic acids.
As inorganic acids, there may be used, for example, hydrochloric
acid, sulfuric acid, phosphoric acid or hydrobromic acid, and as
the organic acids, for example, carbonic, sulfo or sulfonic acids,
such as acetic acid, tartaric acid, lactic acid, propionic acid,
glycolic acid, malonic acid, maleinic acid, fumaric acid, tannic
acid, succinic acid, alginic acid, benzoic acid, 2-phenoxybenzoic
acid, 2-acetoxybenzoic acid, c'nn~m'c acid, mandelic acid, citric
acid, malic acid, salicylic acid, 3-aminosalicylic acid, ascorbic
acid, embonic acid, nicotinic acid, isonicotinic acid, oxalic
acid, amino acids, methanesulfonic acid, ethanesulfonic acid, 2-
hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid, benzene-
sulfonic acid, 4-methylbenzenesulfonic acid, or naphthalene-2-
sulfonic acid.
In the preparation of the drugs for the treatment of the diseases
mentioned, an effective amount of the PDE I inhibitors or salts
thereof is used in addition to the usual excipients, vehicles,
and additives. The dosage depends on the species, body weight,
age, individual condition, and type of administration.
Possible forms of application are oral, intravenous, intra-
-
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-- 4
muscular, subcutaneous, and intralllmln~l (e.g. intrasvesical)
formulations. The latter are, in particular, solutions and
formulations as used for parenteral administration as well.
Formulations for parenteral administration will range from 0.05 ~g
to 100 mg, preferably from 1 mg to 50 mg, of the compounds men-
tioned per unit dose and may be present in separate unit dose
forms, such as ampoules or vials. Preferably, solutions of the
active ingredient are used, more preferably aqueous solutions,
and mainly isotonic solutions, but also suspensions. These
injection forms may be provided as a ready preparation, or they
may be formulated only immediately before use by admixing the
effective compound, ~or example, the lyophilizate, optionally with
other solid carriers, with the solvent or suspension medium
desired.
For oral a~; ni stration, there are used the usual galenic prepara-
tions, such as tablets, coated tablets, capsules, dispersible
powders, granules, aqueous or oily suspensions, syrups, liquors,
or drops.
Solid preparations may contain inert excipients, additives and
vehicles, such as calcium carbonate, calcium phosphate, sodium
phosphate, lactose, starch, mannitol, alginate, gelatin, guar gum,
magnesium or aluminium stearate, methylcellulose, talcum, highly
dispersed silicic acids, silicone oil, higher-molecular fatty
acids (such as stearic acid), gelatin, agar-agar, or vegetable
or animal fats and oils, solid high-molecular polymers (such as
polyethylene glycol); formulations useful for oral administration
may optionally contain additional flavoring and/or sweetening
agents.
Liquid preparations may be sterilized and/or may optionally
contain additives, such as preservatives, stabilizers, wetting
agents, penetration agents, emulsi~iers, spreading agents,
solubilizers, salts ~or adjusting the osmotic pressure or for
buffering, and/or viscosity modifiers.
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Such additives are, for instance, tartrate and citrate buffers,
ethanol, complexing agents (such as ethylenediaminetetraacetic
A acid and its non-toxic salts).
For adjusting the viscosity, there may be used high-molecular
polymers, such as, for example, liquid polyethylene oxide,
carboxymethylcelluloses, polyvinylpyrrolidones, dextranes, or
gelatin. Solid vehicles are, for instance, starch, lactose,
mannitol, methylcellulose, talcum, highly dispersed silicic acids,
higher-molecular fatty acids (such as stearic acid), gelatin,
agar-agar, calcium phosphate, magnesium stearate, animal and
vegetable fats, solid high-molecular polymers (such as poly-
ethylene glycol).
Oily suspensions for parenteral or topical (in this case intra-
vesicular) administrations may contain vegetable, synthetic or
semisynthetic oils, such as, for instance, liquid fatty acid
esters having from 8 to 22 carbon atoms in the fatty acid chains,
for example, palmitic, lauric, tridecylic, margaric, stearic,
arachic, myristic, behenic, pentadecylic, linolic, elaidic,
brassidic, erucic or oleic acids, which may be esterified with
monohydric to trihydric alcohols having from 1 to 6 carbon atoms,
such as, for instance,
methanol, ethanol, propanol, butanol, pentanol, or isomers
thereof, glycol, or glycerol. Such fatty acid esters are, for
instance, commercially available miglyols, isopropyl myristate,
isopropyl palmitate, isopropyl stearate, PEG 6-caprate,
caprylates/caprates of saturated fatty alcohols, polyoxyethylene-
glycerol trioleates, ethyl oleate, waxy fatty acid esters, such
as synthetic duck uropygial fat, coconut oil fatty acid isopropyl
ester, oleic acid oleyl ester, oleic acid decyl ester, lactic acid
ethyl ester, dibutyl phthalate, adipic acid diisopropyl ester,
polyol fatty acid ester, etc. Also useful are silicone oils of
various viscosities or fatty alcohols, such as isotridecyl
alcohol, 2-octyldodecanol, cetylstearyl alcohol or oleyl alcohol,
fatty acids, such as oleic acid. Further, vegeta~le oils, such
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as castor oil, almond oil, olive oil, sesame oil, cottonseed oil,
peanut oil or soybean oil, may be used. The materials mentioned
have the additional property of a spreading agent, i.e. there will
be a particularly good spreading on the skin.
As solvents, gelling agents and solubilizers, there may be used
water or water-miscible solvents, for example alcohols, such as
ethanol or isopropyl alcohol, benzyl alcohol, 2-octyldodecanol,
polyethyleneglycols, phthalates, adipates, propylene glycol,
glycerol, dipropylene or tripropylene glycol, waxes, methyl-
cellosolve, cellosolve, esters, morpholines, dioxane, dimethyl-
sulfoxide, dimethylformamide, tetrahydrofurane, cyclohexanone,
etc.
As film-forming agents, there may be used cellulose ethers which
can dissolve or swell both in water and in organic solvents and
will form a kind of film after drying, such as hydroxypropyl-
cellulose, methylcellulose, ethylcellulose, or soluble starches.
Mixed gelling and film-forming agents are also possible. In this
case, there are chiefly used ionic macromolecules, such as sodium
carboxymethylcellulose, polyacrylic acid, polymethacrylic acid,
and salts thereof, sodium amylopectine semiglycolate, alginic acid
or propylene glycol sodium salt, gum arabic, xanthan gum, guar
gum or carrageen.
As additional formulation aids, there may be used: glycerol,
paraffins having different viscosities, triethanolamine, collagen,
allantoin, novantisolic acid, perfume oils.
The use of surfactants, emulsifiers or wetting agents may also
be required for the formulation, such as, for example, sodium
lauryl sulfate, fatty alcohol ether sulfates, disodium N-lauryl
~-iminodipropionate, polyoxyethylated castor oil, or sorbitan
monooleate, sorbitan monostearate, cetyl alcohol, lecithin,
glycerol monostearate, polyoxyethylene stearate, alkylphenol
polyglycol ether, cetyltrimethylammonium chlori-de, or monoalkyl/
dialkyl polyglycol ether ortho-phosphoric acid monoethanolamine
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salts.
Stabilizers, such as montmorillonites or colloidal silicic acids,
for the stabilization of emulsions or for preventing decomposition
of active substances, such as antioxidants, for example, toco-
pherols or butylhydroxyanisol, or preservatives, such as
p-hydroxybenzoic acid ester, may also be required for the pre-
paration of the formulations desired.
For promoting penetration, intravesicular formulations preferably
contain highly compatible organic solvents, such as ethanol,
methylpyrrolidone, polyethylene glycol, oleyl alcohol, octanol,
linolic acid, triacetin, propylene glycol, glycerol, solketal,
or dimethylsulfoxide.
The preparation, filling and sealing of the preparations is done
under the usual antimicrobial and aseptic conditions. For topical
or transdermal application as well, the preparations are
preferably packed in separate unit doses for easy handling, if
required for stability reasons, as with parenteral forms, also
by separately packing the active ingredients or their combinations
as lyophilizates, optionally with solid carriers, and the solvents
required etc.
Exam~le 1 - Iniection
Fifty milligrams of vinpocetine are dissolved in distilled water
together with 750 mg of NaCl, the pH is adjusted to 3.7 with 1 N
HCl, distilled water is added to give a total of 100 ml, and the
solution is packed in 0.5 ml ampoules.
Exam~le 2 - Solution for To~ical Administration
From 500 mg of vinpocetine, 2 ml of isopropyl myristate and 10
ml o~ ethanol, a solution for topical administration is prepared
and packed in unit doses o~ 2 ml each.
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The efficacy and potency of the drugs according to the teaching
of the invention is demonstrated by the following pharmacological
studies: -
Human urinary bladder muscles freshly collected in the course of
an operation is cut into small strips (about 3 x 3 x 6 mm). The
latter are then installed in an organ bath containing a nutrient
solution ensuring survival of the organic strips. By coupling the
organic strips to a measuring element, length and force changes
of the organic strip can be recorded, and thus actions of
compounds added to the organ bath nutrient solution can be
examined through the length and force changes (increase or
decrease) of the organic strip. At the beginning of the experi-
ment, the organic strips are contracted with an appropriate
standard compound (e.g., carbachol). After the contraction of the
organic strips is completed, an inhibitor of phosphodiesterase I
is now added in incremental dosage (10-9, 10-8, 10-7 etc. mol/l) to
the organ bath solution, and the relaxation triggered thereby is
measured. The results obtained are essentially applicable to the
whole organism since human tissue had been used and the metabolic
processes studied proceed faster in the whole organism and thus
the compounds will act even more pronounced.
Figures 1 to 6 illustrate the results of these organ bath
experiments.
Figure 1 shows the relaxing effect of cumulatively increasing
concentrations of papaverine, an non-specific phosphodiesterase
inhibitor, on human detrusor strips precontracted with 1 ~M
carbachol. The curve shows the average values of measurements each
performed on 12 detrusor strips. This no drug, because of its
severe side-ef~ects due to interaction with all PDEs.
Figure 2 shows the relaxing effect of cumulatively increasing
concentrations of vinpocetine, an inhibitor o~ PDE I, on human
detrusor strips precontracted with 1 ~M carbachoL The curve shows
the average values of measurements each performed on 12 detrusor
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g
strips.
Figure 3 shows the relaxing effect of cumulatively increasing
concentrations of milrinone, an inhibitor of PDE III, on human
detrusor strips precontracted with 1 ~M carbachol. The curve shows
the average values of measurements each performed on 10 detrusor
strips.
Figure 4 shows the relaxing effect of cumulatively increasing
concentrations of rolipram, an inhibitor of PDE IV, on human
detrusor strips precontracted with 1 ~M carbachol. The curve shows
the average values of measurements each performed on 8 detrusor
strips.
Figure 5 shows the relaxing effect of cumulatively increasing
concentrations of zaprinast, an inhibitor of PDE V, on human
detrusor strips precontracted with 1 ~M carbachol. The curve shows
the average values of measurements each performed on 10 detrusor
strips.
Figure 6 shows the relaxing effect of cumulatively increasing
concentrations of dipyridamole, an inhibitor of PDE V, on human
detrusor strips precontracted with 1 ~M carbachol. The curve shows
the average values of measurements each performed on 10 detrusor
strips. Inhibitors of PDE II have not been known to date and
therefore could not be tested.
The proof as to whether a compound is suitable for the purpose
according to the invention, i.e. is an inhibitor of PDE I, is
furnished by known methods, such as described by Galwan et al.,
Arch. Pharmacol. 1990, 342, 221-227; or Nicholson, Br. J.
Pharmacol. 1989, 79, 889-897; for example, according to the
following general procedure:
Fresh tissue obtained during an operation is homogenized and then
ultracentri~uged. Next, the supernatant is filtered, pipetted off
and chromatographed. The determination of PDE isoenzymes is
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- 10 -
performed as described in M. Truss et al.: Urology 45(5): 893-901,
1995. The determination of the amount of radioactivity permits
to calculate the enzyme activity in pmol/ ml/min. A plot of the
activity curve allows to identify fractions in which the phospho-
diesterase activity is particularly high. The phosphodiesterase
activity of each of the peaks exhibits a different composition
with respect to the activity of the different substrates. This
special composition of the phosphodiesterase activity allows for
the assignment to a phosphodiesterase isoenzyme (PDE). A substance
is considered as PDE inhibitor lf the concentration thereof which
is necessary for inhibiting 50~ of the substrate hydrolysis (IC50)
is at least 20 times lower in the respective peak fraction con-
taining the correspoonding phosphodiesterase isoenzyme than in
other peak fractions. For this purpose, enzyme preparations are
again carried out, as described above. Now, however, the compound
to be tested is added prior to the incubation of the enzyme
mixtures according to peak fractions. Then, renewed determination
and plotting of the enzyme activity allows to identify a substance
as being an inhibitor of the phosphodiesterase isoenzyme according
to the above-mentioned definition.
