Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
This applicati~n is a division ~f Canadian
Application No. 389,484 filed November 5, 1981 for
Pharmaceutical Composition ~or Injection.
The present invention relate~ to pharmaceutical
preparations comprising9 as a main ingredient, 4-carbamoyl-
imidazolium-5-oleate or its salt or hydrate thereof (here-
inafter referred to as "Compound A") and to a method for
the productio~ thereof.
It has been known that the Compound A can be
prepared by the method described, for example, in J. Am.
Chem. Soc. Vol. 74, 2892 (1952), and that the Compound A i~
useful as a therapeutic agent for the treatment of tumor,
rheumatism and nephritis (U.S. Patent No. 4,181,731;
French Patent No. 77271~4; Canadian Patent No. 1,078,736).
It is also known that the Compound A can be formu-
lated into various dosage forms such as tablets, capsules,
and injections.
It has now been foundg however, that the Compound
A is so unstable that it is easily colored upon expo~ure
to oxygen, heat, or light, and that, when the Compound A
i9 formulated into pharmaceutical preparations such as
in~ections, oral dosage forms or suppository, the coloration
o~ the ~ompound A i ~urther increased presu~ably due to the
interaction of the Compound A with some other components
such as solvents, diluents or baseq t}~ough co~plicated
reaction.
In addition, the Compound ~ has a poor solubility
in water ~about 4.7 mg/ml at ?5~ as free base form3.
Thi~ poor solubility o~ the oompound creates difficultie~
in the prsduction of in~ections of the compound in that the
~olumes o~ the in~ection~ become too large to be practical
for intra~enous or intramu~cular in~ection, local ln~ection
to tumor and the like.
_ 1 _
;9~
In order to overcome above mentioned difficultles9
we have extensively studied, and as the results, have found
that sulfur compolnd~ capable of producing ~S03 , S032
or S2052 ions are very effective for the control or
prevention of coloration. A~ shown in the Experimental
Example 1 below9 various stabilizers were examined in
expectation of anticoloration by their antioxidative effectR
or chelating of heavy metal ions therewith, but the material~
tested proved to have no e~fect or even increase said colora-
10 tion. ~e have found that the sulfur compounds capable of
producing ~S03 , S032 or S2052 ions can effectively
prevent the coloration of the co~pound. ~en the prepara-
tions containing said sulfur compounds were prepared and
tested, it was found that excellent stabilization effect
was realized in addition to the anti-coloration effect.
For further improvement in a long period stabi-
lity thereo~, studies were continued and the following wa~
found out. That is, an excellent anti-coloration effect
~as obtained by the combination of sodium metabisulfite
20 and ~-cysteins hydrochloride. ~his was indeed a surpris-
ing finding, because the effect is synergistic only in this
particular combination. Furtherm~re, an aqueous injection
of the compound in combination with sodium metabisulfite
and ~-cysteine hydrochloride did not show any coloration
after storing at a room temperature for a long period of
time even under severe conditions, as shown in Experimental
~xample 5.
From these facts~ the inventors have also found
that by the addition of sulfur compound capable of produc-
30 ing one ions selected from HS03 9 S032 and S2052 in com-
- 2 -
bination with ~-cysteine hydrochloride to the compound,
it is possible to obtain a stable preparation showing
no coloration even storing for a long period of time)
and have succeeded in attaining the anti-coloration and
stabilization in full.
Though the invention has been explained in con-
nection with aqueous injection which is believed to be most
difficult to be stabilized, the invention can likewise be
applied to various other pharmaceutical preparations includ-
ing oral dosage form, o.intment, suppository and the like.
Examples of said sulfur compounds include bisul-
fites (e.g. alkali metal bisulfite as sodium bisulfite
~Ta~S03~, potassium bisulfite (KHS03) or the like, and
ammonium bisulfite (NH4HS03)), aqueous sulfurous acid or
sulfites (e.g. alkali metal sulfite as sodium sulfite
(~a2SO~), potassium sulfite (K2S03) and the like, and
alkaline earth metal sulfite as calcium sulfite (CaS03),
barium sulfite (BaS0~) and the like), metabisulfites (e.g.
alkali metal metabisulfite as potassium metabisulfite
(E2S205~, sodium ~etabisulfite (Na2S205)) and the likeO
A~ to the weight ratio of sulfur compound capable
of producing ions selected from HS03 , S032 and S2052
and L-cysteine hydrochloride in the prepar~tion of thi~
invention, it is preferred ~rom the standview of stabili~a-
tion effect de~ired and safety use of additives, to use
0.001 to 0.500 part of the sulfur compound and 0.01 to
0.50 part of ~-cysteine hydrochloride per one part of the
Compound A.
In ca~e of injection, a daily dose of the Compound
A is 50 to 2000 mg for an adult person
-- 3 --
Next, studies have been continued to find out
means to improve solubility of the compound. Various
surfactants usually employed as solubilizing means,
additives in anticipation of intramolecular action, and
non-aqueous vehicles have been tried, but failed to attain
the de~ired effect, but very surprisingly, we have found
that marked improvement in solubility of the compound i9
obtained by the addition of basic subs-tance. On the basis
of this finding, improving solubility which is another
object of this invention has been attained in full, thereby
succeeding in providing an injection of the compound with
wide applicability.
The term "basic material" as used herein may
include alkali metal hydro~ides such as NaOH and the like;
alkali metal ca:rbonate such as Na2CO3 and the like; alka-
line earth meta:L hydroxides such as Ca(OH)2 and the like;
compound~ having alkali metal or alkaline earth metal ion
as cation, for example, borax and the like; and organic
amine3 such as ethanolamine, trishydroxymethyl amino
methane, basic amino acid (e.g. ~-arginine, ~-lysine) and
the like. Anyone of the abovesaid basic materials are
successfully used, but from a p~actical sense of view,
organic amines are preferable.
The solubility of the compound tends to be in-
creased with the amount of said basic material added and
the increase in pH. However, at the same time, cares
shoula be paid to the safety i~ connection with the absolute
adding amount of the basic material, and to select optimum
pH range of the injection solution giving no local stimula-
tion and the like. In thi~ regard, the most desirable pH
ran~e of the injection solution is 8.0 to 9.5 and undersuch condition~, the preferable basic material~ are organic
amines because of easiness in handling and quality of
the product obtained. Among them~ the most preferable one
~ arginine since.it ha~ an excellent 301ubility, safety
factor as additive and operational easiness in handling
thereof. If desired, said basic materials may be used
in the form ~f mixture thereof.
Th~ amount of basic material is to be determined
princi.paly by the amount of the active ingredient, the
Compound A and the total quality evaluation of the prepa-
ration thus obtained including safety factor, but prefer-
ably it i~ in a range of 0.9 to 3 ~oles per mole of the
c ompound .
Thus, the present invention makes it possible
to provide pharmaceutical preparation~ of the Compound A
with improved anti-coloration and stabilization effects
by the addition o~ sulfur compound capable of producing
at least one of the ions selected from HS0~ , S032 and
S2052 ; to attain more complete anti-coloration and
stabili~ation e~fects even in the preparation which is
very liable to be colored, and which is unstable like an
aqueous injection by the addition of ~-cysteine hydrochloride
in combination with the said sulfur com?ounds; and to
improve solubility of the compou~d by the addition of
basic material and especially L-arginine, thereby furnish-
ing wide variety of injections with up to higher concent-
ration.
In preparing the present preparations, the above~
said stabili er and/or solubilizer is used depending on
-- 5 --
6~7
the requirements of the respective dosage form and
using other pharmacologically permissible additives as
diluents, bases, solvent or the like as occasion demands,
various dosage forms are formulated following conventional
means.
Unique properties and usefulness of the invention
shall be now explained in the following Experimental
Examples.
perime ~
To an aqueous solution of the Compound A (lO mg/ml)
containing ~-arginlne, was added each of the stabilizers
listed in the following Table l in a concentration of
0.2 %. ~ach 5 ml of the solution was taken in a 8 ml
ampule, and after replacing the remaining air with nitro-
gen gas, the ampule was stored at 50 C. for l week and the
degree of coloration of the content was measured by light
absorbance at 420 nm (layer length lO mm). In Table l,
was also ~iven color which was determined following JIS Z
QlO2-l457 "Color name", Kogyo-yo Shikimeicho, K.K. Nihon
Shikisai Sha by visual inspeotion.
Table
Sample material ~ ght Color
~ _ . ~
control 0.383 pale yellowish
0.2% sodi.um sulfite 0.038 colorles~
0.2~ sodium bisulfite 0.040 colorless
0.2% sodium metabisulfite 0.035 colorless
-- 6 --
Table l (cont'd)
0.2~ ED~A-2Na 0.520 bright yellowish
green
0.2~ L-ascorbic acid 0.092 faint yellowish
green
0.2~ Erysorbic acid 2.7~6 yellow ocher
0.2% thiourea 1.016 light yellowish
green
0.2~ Taurine 0.903 light yellowish
green
0.2~ acetylthiourea 0.726 light yellowish
green
Experimental Example 2
-
Freeze~dried preparation containing 20 mg/vial of the
compound A alone (control) and together with 10 mg/vial of
sodium bisulfite were prepared. After storing at 40C. or
50 C~ under light shielding condition or room temperature under
1000 Lux, for 2 months, the degree of coloration was measured
by visual inspection and shown in Table 2, in conformity to
JIS Z 8102-1957 "Color name", Kogyoyo Shikimeicho, K. K. Nihon
Shikisai Sha.
Table 2
Storing condition
Sample Light Light 1000 Lux
20material shielding shielding room temp.
at 40C. at 50C.
.
control white - faint faint faint
bluish purple bluish purple bluish green
10 mg sodium white white white
bisulfite
~x~erimental Exam~le 3
Freeze dried preparation containing 100 mg
of the Compound A, 200 mg of I,arginine as solubilizer
and 5 mg of sodium bisulfite per vial and being placed
in a vial, the inner air of which ~as replaced with
nitroge~ gas, was prepared and after storing at 503C.
under light shielding condition for 2 months, the conte~t
of the compound was measured by using light absorbance
ratio at 276 nm. The results are shown~in Table 3.
Table
Sample material ~-oleate originall idazolium
co~trol (not added with
10 sodium bisul~ite) 89
added with 5 mg of
sodium bisul~ite 95
~xperimental EXample 4
. . _
To an aqueous solution of the Compound A
(10 mg/ml) containing I,arginine~ various stabilizers
were added each alone or in the combination formt in
an amount o~ 0.2 %. Each 5 ml of these solutions wer~
taken in the respective 18 ml vial, applied with
stopper, stored at 50C. for 24 hours, and the degree
of coloration was measured by light absorbance at
420 nm (layer length 10 mm). A1SO, the final color
~Jas determined in conformity to JIS Z 8102-1957 "Color
name", Kogyoyo Shikimeicho, K.K. I~Tihon Shikisai Sha,
by visual inspection.
Table 4
Sa~ple material ~ight Color
control 2.392 golden color
C.2~ sodium sulfite 1.472 bright yellow
0.2~ sodium bisulfite 1.120 bright yellow
0.2~ sodium ~etabisulfite 1.532 bright yellow
0.2~ ~DTA-2Na 1.832 golden color
0.2% ~-ascorbic acid 1.582 dark yellow
0.2~ ~-cysteine hydro-
chloride 1.683 golden color
0.2% sodium metabisulfite
plus 0.2% EDTA-2Na 0.833 brlght yellow
0.2~ sodium metabisulfite
plu3 O. 601 pale yellow
0.2% l-ascorbic acid
0.2~ sodium metabisfulfite
0.2% ~-cysteine hydroch- 0.286 faint yellow
loride
~xperlmental_ ~
Aqueous injection each containing the following
in indicated amount~ per ampule were prepared and stored
under severe condition of 50C., 1000 ~UX 9 or at room tem-
perature, and the degree of coloration ~as determined by
measuring light absorbance at 420 nm (layer length 10 ~m),
and compared with each other. In this example, in prepar~
ing the injection solution, oxygen-free water was used
and the inner space of the ampule was replaced with nitrogen
gas.
_ g _
Prescription (per æm ule~
4-carbamoyl-imidazolium-5-oleate100 mg
~-arginine 200 mg -
sodium metabisulfite 4 mg
~-cysteine hydrochloride 10 mg
benzyl alcohol 50 mg
sterilized distilled water
for injection to make 5 ml
Tab e 5
Storing condition
~lght absorbance
initial 0.020
50Co - 2 months 0.002
1000 ~ux 2 months 0.007
room temp. - 6 months 0.015
Experimental Exam le 6
Excess amounts of the Compound A were dispersed
in aqueous solutions containing various surfactants and
additives, respectively. In each case, solubility of the
compound was determined by measuring li~ht absorbance at
277 nm with the filtrate. For non-aqueous vehicles marked
with *, the corresponding solubility of the compound was
judged by eye-measurement.
Ta~le 6
Solubility Solubility
mg/ml of 4- mg/ml o~ 4-
Solvent carbamoyl- Solvent carbamoyl-
imldazolium- imidazolium-
5-oleate 5-oleate
~ _ . _ _ . _ .
5% Amycol No. 1
water 4 7 (manufactured
by Nichiden 5.1
Kagaku K.K.)
5~o polyoxyethy-
nated YCaStgr oil 4 5 deoxycholate white gel
5% polyoxyethy-
lene sorbitan 4.6 saturated glu-
monooleate tamic acid 5-3
5% sod um lauryl 4 7 saturated aspartic 5.4
* ethanol less than 2 saiUdrated gentisic whit
* propylene
glycol about 5 5C~ levul.inic acid 6.
~o * polyethylene less
glycol than 2 5~o tartaric acid 7.0
* glycerine about 3 lOC/o mannitol 4.6
* N,N-dimethyl- less
acetamide than 2 5,~ sodium salicylate 6.5
* sesame oil less 5~o nicotinic amide 7.2
than 2
* cotton seed_ less 5~ ethylurea 5.6
* corn oil less
than 2
~e~ac~
Using the same procedures as stated in EXperi-
mental Example 6~ solubility measurement was carried out.
11 -
,
~able 7
. .
Solubilit~ of 4-
carbamoyl-imi- PH of
Solvent dazolium-5- solvent
oleate, mg~ml
1% ethanolamine 24~9 8.64
50thrishydroxy methylamino 25-7 8.56
5% Irarginine 33.5 8.69
5% Irlysine 10.7 8.20
0.5% NaOH 21.4 8.50
NaHC03 Na2C03 buffer 26.7 8.42
5% borax 22.2 8.44
The invention shall be now more fully explained
in the following examples, which, however, should not be
taXen as being limitative in any sense.
Example
A mixture of 4-carbamoyl-imidazolium-5-oleate
(100 g), sodium bisulfite (5 g) and l,arginine (200 g)
was dissolved in distilled water for injection to give the
total ~olume 10 Q of the solution. The solution was fil-
tered to remove bacteria, filled in each 10 ml into vials
(24 ml in volume) and freeze-dried (the inner space of vial
being replaced with nitrogen gas) to obtain freeze-dried
injection, which was stable and capable of being reconstituted
in higher concentration in less than 10 ml of distilled water
for injectio~ before use.
- 12 -
~B~i~7
Exam~le 2
~ he same procedures as stated in Example 1
~rere repeated, excepting substituting sodium sulfite for
sodium bisulfite, to obtain freeze-dried injec~ion, which
was stable and capable of being reconstituted in higher
concentration in less than 10 ml of distilled water for
injection before use.
~ he sa~e procedures as stated in ~xample 1
were repeated, excepting substituting sodium metabisulfite
for sodium bisulfite, to obtain freeze-dried injection~
which was stable and capable of being reconstituted in
higher concentration in less than 10 ml of distilled ~ater
for injection before use.
Exa,m~ e,,~
A mixl,ure of 4-car'bamo~l-imidazolium-5-oleate
(100 g), sodium bisulfite (2.5 g), sodium metabisulfite
(2.5 g) and I,arginine (200 g) was dissolved in distilled
water for injection to give the total volume '0 ~ of the
solution. The solution was filte~ed to remove b~cteria,
filled in each 10 ~1 into vials (24 ml in vol-lne), and
subjected to ~reeze-drying to obtain freeze-dried injec-
tion being stable and capable of being reconstituted inhigher concertration in les~ than 10 ml of distilled
~ater for injection before use.
, - 13 -
156 g of 4-carbamoyl-imidazolium-5-oleate
hydrochloride, 3.1 Q of 1 % aqueous sodium hydroxide solu-
tion and 200 g of L-arginine were dissolved, under stirring,
in distilled water for injection, to make the total volume
to 10 liters. This was filtered to remove bacteria, filled
in each 10 ml into vials (24 ml i~ volume) and subjected
to freeze-drying -to obtain freeze-dried injection being
stable and capable of being reconstituted in higher con-
centration in less than 10 ml of distilled water for
injection before use.
Exam~le 6
into 5 liters of oxygen-free, sterilized, distilled
water for injection, were dissolved gently 4 g of sodium
metabisulfite, 10 g of L-cysteine hy~rochloride and then
100 g of 4-carbc~moyl-imidazolium-5-oleate, 200 g of ~-
arginine and 50 g of benzyl alcohol. Thus obtained solution
was filtered under sterile condition, each 5 ml of which was
then filled into ampules (5 ml in volume), and after replac~
ing air in vessel with nitrogen gas, the a~pule was sealed
to obtain an aqueous injection of 4-carbamoyl-imidazolium-
5-oleate which was stable for a longer period of time.
Compound ~ 2000 g
crystalline cellulose 2400 g
ma~nesium stearate 50 g
sodium bisulfite 150 g
The above-mentioned ingredients were ~ixed
together a~d thus obtained powdery mixture was formed
into tablets each weighing about 230 mg. This was
proven to be very stable for a longer period of time.
B ample 8
Compound A 400 g
milk sugar 600 g
sodium metabisulfite 20 g
l-cysteine h~drochloride 40 g
~ he above-mentioned ingredients were mixed
together to obtain uniformly mixed powder, which was
10 stable for a longer period of time.
- 15
