Note: Descriptions are shown in the official language in which they were submitted.
HA160a
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Method of Treat ng Hypertension and
Medicaments Thereor
; The present invention relates to a method for
reducing or alleviating hypertension with a combination
comprising an effective amount of a compound of the
formula
(I)
4 ~1 ~
~ R~-S-(CH)n- CH- CO- N _ COR
: wherein R is hydroxy, lower alkoxy or NH2;
; Rl and R4 each is hydrogen, lower
alkyl ox phenyl-lower alkyl;
R2 is hydrogen or R5-CO;
R3 is hydrogen, hydroxy or lower alkyl;
R5 is lower alkyl, phenyl or phenyl-
lower alkyl; and
n is 0, 1 or 2.
with an effective amount of a diuretic compound and
such a combination of medicaments~
.~
l~ZV~
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The compounds of formula I have been reported
to be angiotensin converting enzyme inhibitors which
intervene in the angiotensinogen ~ renin ~
angiotensin I ~ angiotensin II mechanisn and are
effective in reducing or alleviating hypertension.
See U.S. Patent 4,046,889, September 6, 1977;
Science 196, 441-443 (1977). It has been found that
such compounds can be used in an oral dosaqe range
of about 0.1 to 100 mg/kg per day and are most
effective when provided at a total daily dosage of
about 60 to 600 mg. Dosages within this range achie~e
a substantial reduction in arterial blood pressure
and, in most instances, little, if any significant
reduction is obtained by further increasing the
dosage. Although certain peptides, teprotide (SQ20,881)
for example, have ~een reported to have angiotensin
converting enzyme activity, they are not of practical
use for such an indication because of the cost
and particularly since they are ineffective when
orally administered [Rubin et al., 204, Jour. Pharm.
Exper. Ther. 271-280, 1978; Laffan et al., Jour.
Pharm. Exper. Ther. 204, 281-288, 1978; Brit. Med.
Jour. 2(6141~:866, 1978].
Hypertension is also frequently treated by the
administration of a diuretic. Typically, treatment
with an antihypertensive agent alone results in a
compensatory retention of sodium and water which
concomitant administration of a diuretic
prevents (Wollam et al., Drugs 14:420-460, 1977).
However, administration of a compound of
formula I does not result in sodium and water
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retention when administered alone and, in fact, may
by itself cause natriuresis and diuresis (Bengis et al,
Circulation Research, Vol. 43 I-45-I-53, 1978).
Therefore, a diuretic woul~ not be expected to enhance
the antihyper~ensive action of compounds o~ formula I.
However, it has been demons~rated that the administra-
tion of a diuretic in combination with compounds of
formula I is more effective than either drug alone.
The combination of such compounds with a diuretic as
described below results in a potentiation of the
reduction in blood pressure significantly beyond that
level which either substance can achieve itself at a
dosage within the acceptable range and also at
lower dosage levels.
This invention therefore relates to a combination
of a compound having formula I above and a diuretic
of the group consisting of the thiazide class, e.g.,
chloro~hiazidel hydrochlorothiaæide, flumethiazide,
hydro1umethiazide, bendroflumethiazide, methychlo-
thiazide, trichlorme~hiazide, polythiazide or
b~nzthiazide, as well as ethacrynic acid, ticrynafen,
chlorthalidone, furosemide, bumetanide, triamterene,
amiloride and spironolactone, and salts of such
- compounds, compositions comprising a combination of
such compounds and to a method for alleviating hyper-
tension with a combination of compounds.
Preferred are those compounds of formula I
wherein R is hydroxy or lower alkoxy, especially
Cl~C4 lower alkoxy; Rl is hydrogen or lower alkyl,
especially methyl, R2 is hydrogen or lower alkanoyl,
especially C2-C4 lower alkanoyl; R3 is hydrogen
'~ ~ ~A160a
or hydroxy/ especially 4-hydroxy; R~ is hydrogen or
lower alkyl, especially Cl-C~ lower alkyl; and n is
0 or 1. Especially pre~erred in this group are com-
pounds of Eormula I wherein ~ is hydroxy; Rl is
hydrogen or methyl; R2 is hydrogen or acetyl; R3 is
hydrogen; ~4 is hydrogen or methyl; and n i5 0
or 1. The especially preferred embod:iment includes
a compound of formula I wherein R is hydroxy; Rl is
methyl; R2, R3 and R4 each is hydrogen; and n is 1,
most especially tD-3~mercapto-2-methyLpropanoyl)-L
proline.
Preferred as the second component of the
combination is chlorothiazide, hydrochlorothiazide,
furosemide, ticrynafen or triamterene, especially
1~ hydrochlorothiazide or furosemide.
The especially preferred embodiments are
compositions comprising (D-3-mercapto-2-methvl-
propanoyl)-L-proline with either hydrochlorothia~ide
or furosemlde.
~ ~0 The compounds of formula I can be produced as
- described in U.S. Patent 4,046,889, September 6,
1977. The diuretic members of the combination are
known compounds which are produced by methods
described in the literatureO
According to this invention, a combination of
a compound of formula I and a diuretic is administered
in an effective amount which comprises a total
daily dosage of about 30 to 600 mg., preferably
30 to 300 mg. of a compound of formula I and about
15 to 300 mg. preferably 15 to 200 mg. of the diuretic
to a mammalian species which has elevated blood
~ ~IA160a
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pressure. Such total daily dosages can be used in a
single administration of the total amount or in
divided doses two to four times dai~y. Generally,
a t.i.d. or q.i.d~ regimen is preferred. This pre
5 ferred dosage is about 10 to 100 mg. of the compound
of formula I and about 5 to 75 mg. o~ the diuretic
three times daily or about 5 to 125 mg. of the compound
of formula I and about 2.5 to 50 mg. of the diuretic
four times daily. The preferred route of admini~tration
is oral.
According to one preferred embodiment, the
substances can be formulated in a single pharmaceu-tical
dosage form for oral administration such as tablet,
capsule; solution or suspension comprising an effective
lS amount of each of the active ingredients in a physiolo-
gically acceptable carrier therefor.
The active substances in the dosage unit are
present in a ratio of about 1:2 to about 12:1,
preferably about 2.5:1 to about 10:1, of the compound
of formula I with respect to the diuretic ~by weigh~).
Generally, about 10 to 200 mg. of a compound o~
formula I and about 2.5 to 100 my. of the second
component can be readily formulated in the composition.
Tablets of various sizes can be prepared, e.g.,
2~ of about 50 to 700 mg. in total weight, containing the
active substances in the ranges described above,
with the remainder being a physiologically acceptable
caxrier or other materials according to accepted
pharmaceutical practice. These tablets can~ o~
course, be scored to provide for fractional doses.
Gelatin capsules can be similarly formulated.
~A160a
Liquid formula~ions can also be prepared by
dissolving or suspending khe combination of active
substances in a conventional liquid vehicle acceptable
Eor pharmaceutical administration so as ~o provide the
desired dosage in one to four teaspoonsful.
Such dosage forms can be administered to the
patient on a regimen of one to four doses per day.
According to another modifica-tion, in order to
more finely regulate the dosage schedule, the substances
may be administered separately in individual dosage
uni~s at the same time or carefully coordinated times.
Since blood levels are built up and maintained by a
regulated schedule of administration, the same result
is achieved by the simultaneous presence o~ the two
substances. The respective substances can be
individually formulated in separate unit dosage
forms in a mannex similar to that described above.
~ ixed combinations of the compound o~ formula I
and the diuretic are more convenient and are preferred,
expecially in tablet or capsule form for oral
administration.
In formulating the compositions of this invention
the active substances, in the amounts described above,
are compounded according to accep~ed pharmaceutical
practice with a physiologically acceptable vehicle,
carrier, excipient, binder, preservative, stabilizer,
flavor, etc., in the particular type o~ unit dosage
form.
Illustrative of the adjuvants which may be
incorporated in tablets are the following: a binder
such as gum tragacanth, acacia, corn starch or gelatin;
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an excipient such as dicalcium phosphate or cellulose;
a disintegra-ting agent such as corn starch, potato
starch, alginic acid or the like; a lubricant such
as stearic acid or magnesium stearate; a sweetening
agent such as sucrose, lactose or saccharin; a flavor-
ing agent such as orange, peppermint, oil of
wintergreen or cherry. ~hen the dosage unit form is
a capsule, it may contain in addition to materials of
the above type a liquid carrier such as a fatty oil.
Various o-ther materials may be present as coatings or
to otherwise modify the physical orm o the dosage
unit. For instance, tablets or capsules may be
coated with shellac, sugar or both. A syrup or elixir
may contain the active compound, water, alcohol or the
like as the carrier, glycerol as solubilizer, sucrose
as sweetening agent, methyl and propyl parabens as
preservatives, a dye and a 1avoring such as cherry
or orange.
Many of the active substances described above
2V form commonly known, pharmaceutically acceptable
salts such as alkali metal and other common basic
salts or acid addition salts, etc. References to the
base substances are therefore intended to include those
common salts known to be substantially equivalent to
the parent compound.
The following examples are illustrative of the
invention and constitute especially preferred
embodiments. They also serve as models for the
preparation of ~ther members o the group which
can be produced by suitable substitution of
lngredients as described above.
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Example 1
6000 tablets each containing the following
ingredients:
(D-3-mercapto-2-methylpropanoyl)-
L-proline 100 mg.
*Avicel (microcrystalline cellulose) 100 mg.
Hydrochlorothiazide 12.5 mg.
Lactose U.S.P. 113 mg.
Corn starch U.S.P. 17.5 mg.
Stearic acid U.S.P. 7 mg.
350 mg.
are produced (from sufficient bulk quantities) by
slugging the (D-3-mercapto-2-methylpropanoyl)-L-
~ proline,*Avicel and a portion of the stearic acid.
- 15 The slugs are ground and passed through a ~2 screen,
- then mixed with the hydrochlorothiazide, lactose,
corn starch and remainder of the stearic acid. The
mixture is compressed into 350 ~g. capsule shaped
tablets in a tablet press. The table~s are scored
for dividing in half.
Example 2
10,000 tablets each containing the following
ingredients:
(D-3-mercapto 2-methylpropanoyl)-
25 L-proline 200 mg.
Corn starch U.S.P. 17.5 mg.
Lactose U.S.P. 215.4 mg.
Acacia U.S.P. 10.6 mg.
Water qs (ca. 0.03 ml.~
30 Hydrochlorothiazide 25 mg.
Corn starch U.S.P. 17.5 mg.
* Trade Mark
HA160a
_g_
Avicel 200 mg.
Stearic Acid 14 mg.
700 mg.
are produced from sufficient bulk quantities as
follows:
The acacia is dissolved in wa-terO 17.5 mg.
of corn starch~ the (D-3-mercapto-2-methylpropanoyl)-
L-proline and lactose are mixed thoroughly. The dry
mixture is granulated using ~he aqueous solution of
acacia. The granulation is wet screened, dried at
120F~ and reduced. Th~ reduced, dry granulation is
mixed with the hydrochlorothiazide and the remaining
excipients are then added and mixed. The mixture i5
compressed into tablets of 700 mg. each.
Example 3
Tablets each containing the following ingredients
are made as descrlbed in Example 2:
(D-3-mercapto-2-methylpropanoyl)~
L-proline 75 mg.
Corn starch U.S.P. 8 mg.
Lactose U.S.P. 120 mg.
Acacia U.S~P. 6 mg.
Water qs. (ca. 0.03 ml.
Chlorothiazide 50 mg.
Corn starch U.S.P. 8 mg.
Avicel 75 mg.
Stearic acid 8 mg.
350 mg.
Exam~le ~
1000 capsules, each containing the following
ingredients:
HA160a
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(D-3-mercapto-2-methylpropanoyl)-
L-proline lO0 mg.
Lac~ose U.S.P. 211.8 mgO
Magnesium stearate 3.2 mg.
Hydrochlorothiazide 10 mg.
325 mg.
are produced by dry blending the hulk materials
(except the magnesium stearate) in a Hobart mixer,
then passing the blend through a #20 screen. The
materials are mixed again in the Hobart mixer with
the magnesium stearate. The mixture is then filled
into #2 two-piece gelatin capsules.
Example 5
By substituting 10 mg. of furosemide for the
hydrochlorothiazide in Example 4, capsules containing
furosemide and (~-3-mercapto-2-methylpropanoyl)-L-
proline are similarly produced.
Example 6
By following the proceudre of Example 2 but
substituting 20 mg. of furo~emide for the hydrochloro-
thiazide and using 220.4 mg. of lactose, 700 mg.
tablets each containing 20 mg. of furosemide and
200 mg. of (D-3-mercapto-2-methylpropanoyl)-L-
proline are similarly produced.
Example 7
By substituting 10 mg. of furosemide for the
hydrochlorothiazide and using 115.5 mg.iof lactose
in the procedure of ~xample 1, 350 mg. scored
tablats each containing 10 mg. of furosemide and
100 mg. of (D-3-mercapto-2-methylpropanoyl)-~-
proline are similarly produced.
3~ Oa
Exam~
6000 ~cored ~ablets of 400 mg. each and containing
the following ingredients:
(D 3-mercapto-2~methylpropanoyl)-
L-praline 125 mg.
; Corn starch 8 mg.
Lactose U.S.P. 95 mg.
Acacia 7 mg.
Water qs. (ca. 0.03 ml.
; 10 Triamterene 50 mg.
Corn starch U.S.P. 8 mg.
A~icel 100 mg.
Stearic acid 7 mg.
400 mg.
are produced as described in Example 2.
6000 scored tablets of 350 mg. each and containing
the following ingredients:
(D-3-mercapto-2-methylpropanoyl)-
L-proline 100 mg.
~vicel 100 mg.
Triamterene 25 mg.
Lactose U.S.PO 100 mg.
Corn starch U.S.P. 17 mg.
Stearic acid 8 mg.
350 mg.
are produced as described in Example 1.
Example 10
5000 scored tablets of 180 mg. each and
containing the following ingredients:
~D-3-mercapto-2-methylpropanoyl)-
L-proline 10 mg.
.. ~
HA160a
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Avicel 50 mg.
HydrochlQrothiazide 5 mg.
Lactose U.S.P. 101 mg.
Corn starch U.S.P. 10 mg.
Stearic acid 4 mg.
180 mg.
are produced as described in Example 1.
Example 11
By substituting the same amount of ticrynafen ~or
the hydrochloro~hiazide in Example 1, tablets
containing 100 mg. of (D-3-mercapto-2-methylpropanoyl)-
L-proline and 12.5 mg. of ticrynafen are similarly
obtained.
Representative of the results obtained with
combinations of agents of this invention are data
obtained from studies in spontaneously hypertensive
ra-ts and two kidney renal hypertensive rats.
A) In an acute study with spontaneously
hypertensive rats, ten to fourteen week old male
Wistar-Kyoto spontaneously hypextensive rats
(190-210 gm.) of the Okamoto-Aoki strain (obtained
from Taconic Farms, Germantown, N.Y.) were given
food and water ad libitum and intubated according
to the method of Weeks and Jones, Proc. Soc. Exp.
Biol. Med. 104, 646-648 ~1960), to prepare them
for blood pressure and heart rate determination by
implanting indwelling abdominal aortic ca~heters under
sodium pentobarbital anesthesia.
Three weeks later their direct blood pressure and
heart rate were recorded by the method of Laffan
et al., Cardiovasc. Res. 6, 319-324 (1972), modified
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as follows. The signal from the transducer was
digiti~ed in a 10 bit A/D con~erter and input to a
PDP 11/05 computer~ The computer was programmed to
sense and store samples at a rate of 125/sec for
each rat, as well as the number of pressure pulses
~uring 10 sec. of each scan on each rat. These
parameters were averaged and stored as the MBP
(mean blood pressure, mm Hg) and heart rate (beats/min.)
for that time. Data were acquired from each rat every
five minutes. Six such sets o~ data were averaged
to give a mean value repxesenting a 30 minute
sample and this 30 minute figure was stored for
subsequent analysis. Each time a 48 hour cycle was
compleked (or sooner if demanded) the data were
transferred serially to a host computer (PDP 11/40)
for further analysis and the data were printed out
on a Versatec Printer/Plotter ~or at least 16 hours
after each dose.
The spontaneously hypertensive rats were segregated
into four groups of five rats each (except group 3
which included six rats). The ~ollowing was adminis-
tered to the rats in the respective group~:
1. (Control) Agar - 5 ml./kg + agar - 5 ml./kg
2. Water - 5 ml./kg + Compound A - 30 mg./kg
3. Compound F - 50 mg./kg + Agar - 5 ml./kg
** *
4. Compound F 50 mg./kg + Compound A
30 mg./kg
* Compound A = (D-3-mercapto-2-methylpropanoyl)-
L-proline
** Compound F = Furosemide
S~ HA160a
Compound F was suspended in 0.25% agar and
Compound A was in aqueous solution. All substances
were administered by gavage and there was a one
hour interval between drugs. Test resul-ts were
evaluated 2.5 hours after single oral doses.
The followiny resulks were obtained;
TABLE I
Mean Blood Pressure (mm/Hg)
Before 2.5 hours after
sinc~le oral dose
(1) 173 169
(2) 175 158
(3) 184 172
(4) 177 128
15 ^ In thase studies Compound F alone, 50 mg./kg.
p.o., produced a 9.7% decrease in SHR blood pressure.
Compound A alone, 30 mg~/kg., produced 6.5~ decrease
; in blood pressure. The combination of Compound A,
30 mg.~kg., p.o., + Compound B, 50 mg./kg., p.o.,
reduced blood pressure in SHR rats by 27~7~o
B) In chronic studies with renal hypertensive
rats, male rats (]15-150 g.) of the Charles River
Sprague Dawley (COBS-CO) s~rain were anesthPtized with
ether and a silver clip (0.22 m~ i.d.) was placed
on the left renal artery through a flank incision. The
contralateral kidney was left intact (two-kidney
Goldblatt model: 2-K RHR). Each rat was fitted with a
tail cuff for air inflation and a Korotkoff sound
microphone for the detection of arterial pulsa~ion.
An oscilloscope was used for a visual appearance and
disappearance of ~he pulse. Blood pressure measurements
IA16Oa
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were determined after a minimum of six inflations withsystolic pressures observed on a Narco physiograph
manome~er. Blood pressures were determined initially
just prior to dosing and twice weekly at 4 hours after
S dosing.
The number of rats in each group was 15. Single
daily treatments were made by gavage with crossover
treatments as indicated in the table below. The
c~ntrol gro~p received distilled water. Compo~md
was administered in distilled water, 30 mg./kg~
Compound H was administered in 0.25% methylcellulose.
The mean blood pressure ~mm/Hg.) for each group
before dosing and on day 119 (4 hours after dosing)
and the number of survivors on day 120 is shown
in the table~
HAl 6 0a
--16--
S._
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O ~ ~D ~Oi` ~ ~ ,i Ul
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a~ ~ ~ ~ o ~ o
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1 5 ~ ,~
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.
HA160a
-17-
The foregoing data show that on long term
treatment compound ~I shows no significant decrease
in blood pressure. Compound A alone shows approximately
a 10 to 15~ reduction in blood pressure. The
combination dosing with Compound A and Compound H shows
approximately a 30~ reduction in bloocl pre~sure.
~oreover, the combination is the only one howing
a 100~ survivor rate.
~0
:~0
