Note: Descriptions are shown in the official language in which they were submitted.
~7'~
1 This invention relates to an oxygen-transferable
fluorocarbon compound emulsion for injection and per-
fusion, which is used in the lifesaving of a patient
suffering from massive hemorrhage and in the preserva~tion
of internal organs in transplantation and process for
preparing same.
It has already been reported by a number o-f
research people that fluorocarbon compound emulsions
may possibly be used as an artifical blood substitute
for mammals and as a perfusion fluid for preservation
of internal organs to be transplanted 9 particularly
as a substitute infusion fluid capable of transporting
oxygen ~eland C. Clark, Jr., ~. Becattini, and S.
Kaplan: The physiology of synthetic blood., Journal
of Thoracic Cardiovascular Surgery, 60, 757-773 (1970);
R. P. Geyer: ~luorocarbon-polyol artificial blood
substitutes., ~ew ~ngland Journal of Medicine, 289,
1077 - 1082 (1973)~.
These emulsions, however, cannot be assumed
as satisfactory enough for practical use in view of
their pharmaceutical stability and of safety of living
animal body. In order that fluorocarbon compound
emulsions be qualified for practical use as an artificial
blood substitute, it is necessary to develop a preparation
which is sufficiently stable to be kept for a long period
of time without change in particle size.
In fluorocarbon compound emulsions, the size
of particle plays an important role in the to~icity and
efficacy of the ernulsion ~K. Yokoyama, K. Yamanouchi,
M. Watanabe, R. Murilshirna, 'L'. Matsurnoto, T. Hamnno,
~'7Z~
1 H. Okamoto, T. Suyama, R. Watanabe 7 and R. ~aito:
Preparation of perfluorodecalin emulsion, an approach
to the red cells substitute., Federation Proceedings,
34, 1478-1483 (May, 1975) ~. An emulsion of larger
5 particle size is more toxic and shorter in retention
time of the particles in the blood stream. According-
ly, when the fluorocarbon compound emulsion is intended
for use as an artificial blood substitute for saving
the life of a patient suffering from massive hemorrhage,
its average particle size should be 0. 3~ or less in
diameter, preferably 0.2~ or less [Japanese Patent
Application Kokai (~aid Open) ~o. 22612/73].
Besides the particle size, in order that the
fluorocarbon compound emulsion be usable as artificial
15 blood substitute, it is necessary that after being
eliminated from the blood stream, the intravenously
administered fluorocarbon compound must be excre-ted
from the body as rapidly as possible. Some of the
present inventors had previously studied on~the
excretion rate and toxicity of several sorts of
fluorocarbon compound emulsions and, as a result,
found that perfluorocarbon compounds of 9 to 11 carbon
atoms are usable as the material of artificial blood
substitute, particularly perfluorodecalin being the
best of all [K. Yokoyama, K. Yamanouchi, and R.
Murashima: ~xcretion of perfluorochemicals after
intravenous injection of their emulsion., Chemical
Pharmaceutical Bulletin, 23, 1368-1373 ( June, 1975) ~.
In addition, some of the present inventors
3~ had also found tha-t the fine and stable fluorocarbon
c
~L~'7Z~
1 compound emulsion can be prepared from these selected
fluorocarbon compounds of 9 to 11 carbon atoms by
emulsifying said fluorocarbon compounds with the
mixture of egg yolk phospholipids or soybean phos-
pholipids and a small amount of fatty acids of ~to 22 carbon atoms or salts thereof or monoglycerides
thereof [Japanese Patent Application Kokai (Laid Open),
No. 69,219/75].
However, compared with a perfluorotributylamine
emulsion stabilized with a high-molecular-weight
polyoxyethylene-polyoxypropylene copolymer emulsifier
(R. P. Geyer, loc. cit.)~ the above-noted emulsion
stabilized with both phospholipids and fatty acids
is superior in the rate of excretion, bu-t inferior in
15 the stability in circulating blood stream after intra- ::
venous injection, the half life being about two-thirds
of that of the former.
Further, a fluorocarbon compound emulsion
prepared with a high-molecular-weight nonionic surface
20 active agent such as perfluorotributylamine emulsion,
can be used as a mixture of any proportion with the
commercial plasma expanders such as dextran, or
hydroxyethylstarch, or modified gelatin solution,
whereas the perfluorodecalin emulsion described in
Japanese Patent Application Kokai (~aid Open) No.
69,219/75 cannot be used in combination with said
plasma expander because of formation of precipitates
when mixed with the latter. It seems that the
precipitation is due to the destruction of emulsified
particles caused by interaction between the phospholipid~
~0724~L6
1 contained in a high concentration in the emulsion and
the plasma expander, such as dextran or hydroxyethyl-
starch, which is a high-molecular-weight colloidal
substance.
~hen it is intended to use a fluorocarbon
eompound emulsion as an infusion fluid or an artificial
blood substitute to save the life of patient in the
ease of massive hemorrhage, the combined used with
a plasma expander becomes important in order to make
isotonicity, that is, to equalize oncotic pressures of
both colloidal solutions, i.e. the emulsion and the
blood; the fluorocarbon compound emulsion supplies
oxygen, while the plasma expander makes it possible
to maintain the circulating blood volume at a proper
level. Therefore, it is preferable to use a high-
moleeular-weight nonionic surface active agent which
is inactive with the plasma expnader, in preparing
a fluorocarbon emulsion intended for use as an artificial
blood. Although these high-molecular-weight nonionic
surface active agents are effeetive for some fluoro-
carbon compounds such as perfluorotributylamine and
other amine-type fluorocarbons as an emulsifier,
they are not suitable for fluorocarbon compounds of
9 to 11 carbon atoms, such as perfluorodecalin, which
have a high rate of excretion.
Under the circumstances, the present in-
ventors eonducted extensive pharmaceutical investigations
on preparing the emulsions of those fluorocarbons of
9 to 11 carbon atoms represented by perfluorodecalin,
which have a high rate of excretion, which emulsions
1~72~
1 are stable in the circulating blood strcam and a.re
able to mix with the plasma expander without any
destruction of emulsified particles. As a result,
the present invention has now been accomplished.
According to this invention, there is pro-
vided a stable emulsion in a physiologically acceptable
aqueous medium of an oxygen-transferable perfluorocarbon
compound having a particle size of about 0.05 to 0.3,~ ,
which comprises (A) at least one perfluorocarbon com-
pound having 9 - 11 carbon atoms selected *rom the
group consisting of perfluorodecalin, perfluoromethyl-
decalin, perfluoro alkylcyclohexanes having 3 to 5
carbon atoms in the alkyl, perfluoro alkyltetrahydro-
furans having 5 to 7 carbon atoms in the alkyl,
perfluoro alkyltetrahydropyrans having 4 to 6 carbon
atoms in the alkyl, perfluoroalkanes having 9 to 11
carbon atoms; ~B) at least one perfluoro tert-amine
having 9 to 11 carbon atoms selected from the group
consisting of perfluoro tert-alkylamines having 9 to
11 carbon atoms, perfluoro N-alkylpiperidines having
4 to 6 carbon atoms in -the alkyl, and perfluoro N-
alkylmorpholines having 5 to 7 carbon atoms in the
alkyl; a high-molecular-weight nonionic surfactant
having a molecular weight of about 2,000 to 207000; a
phospholipid; and at least one fatty acid compound
selected from the group consisting of fatty acids
having ~ to 22 carbon atoms 9 physiologically acceptable
salts and monoglycerides thereof; the ratio of the said
perfluorocarbon compound and the said ~erfluoro-tert-
~0 amine being 95 - 50 to 5 - 50 by weight.
~T 5
~)7~
the sum of the concentration of the perfluorocarbon compound (A) and
the perfluoro tert-amine (B) belng 10 to 50% (W/V), the concentration
of the high molecular weight nonionic surfactant being 2.1 to 5.0%
(~/V), the concentration of the phospholipid being 0.1 to 1.0% (W/V) and
the concentration of the fatty acid compound being 0.004 to 0.1% (W/V).
- 5a -
~07Z~
l The "high-molecular-weight nonionic surfactant",
as herein referred to, has a molecular weight of 2,000
to 20,000 and includes polyoxyethylene-polyoxypropylene
copolymers, polyoxyethylene alkyl ethers, and poly-
oxyethylene alkyl aryl ethers. The concentration of
said surfactant in the emulsion is about 2.0 to about
5.0, preferably 3.0 to 3.5, ~o (W/V).
The symbol "% (W/V)" referred to in the
specification and claim of this application means
the amount proportion of a material by weight (gram)
based on lO0 ml of the resulting emulsion.
Examples of the perfluorocarbons (A) having
9 to ll carbon atoms are a perfluorocycloalkane or
perfluoro alkylcycloalkane which includes, for example,
perfluoro C3_s-alkylcyclohexanes such as perfluoro-
methylpropylcyclohexane, perfluorobutylcyclohexane,
perfluorotrimethylcyclohexane, perfluoroethylpropyl-
cyclohexane, perfluorodecalin and perfluoromethyl-
decalin; a perfluoro C4_6-alkyltetrahydropyran such as
perfluorohexyltetrahydropyran; a perfluoro Cs_7-
alkyltetrahydrofuran such as perfluoro pentyltetra-
hydrofuran, perfluoro hexyltetrahydrofuran and per-
fluoro heptyltetrahydrofuran; and a perfluoroalkane
having 9 - ll carbon atoms such as perfluorononane
and perfluorodecane.
Examples of the perfluoro tert-amine (B)
having 9 to ll carbon atoms are a perfluoro tert-
alkylamine having 9 to ll carbon atoms which includes,
for example, perfluorotrialkylamines such as perfluoro
~0 N,N-dibutylmonomethylamine, perfluoro N,N-diethyl-
-- 6 --
~C~72~
1 penty]amine, perfluoro N,N-dlethylhexylamine, per-
fluoro N,N-dipropylbutylamine and per~luorotripropyl-
amine; a perfluoro N,N-dialkylcyclohexylamlne having
9 - 11 carbon atoms such as perfluoro N9N-die-thyl-
cyclohexylamine; a perfluoro N-C~_6~alkylpiperidine
such as perfluoro N-pentylpiperidine, perfluoro ~-
hexylpiperidine and perfluoro N-butylpiperidine; and
~a perfluoro ~-C5_7-alkylmorpholine such as perfluoro
N-pentylmorpholine, perfluoro ~-hexylmorpholine and
perfluoro N-heptylmorpholine.
The ratio of the perfluorocarbon compound
(A) to the perfluoro tert-amine (B) to be used is
50 - 95 to 50 - 5 by weight and the total amount of
(A) and (B) contained in the emulsion is about 10 to
about 50 % (W/V).
The phospholipids used as emulsifier adjuvant
in the invention are ones commonly used in the art,
and those comprising yolk phospholipid or soybean
phospholipid are preferable. The amount present in
the emulsion ranges from about 0.1 to about 1.0 ~0 (W/V),
and preferabIy about 0.~ to about 0.6 % (W/V).
The fatty acid compound used as emulsifying
adjuvant is a fa-tty acid having 8 to 22 carbon atoms,
a physiologically acceptable salt such as sodium or
potassium salt or a monoglyceride thereof, ~hich in-
cludes, for example, caprylic acid, capric acid, lauric
acid, myristic acid, palmitic acid, stearic acid,
behenic acid, palmitoleic acid, oleic acid, linoleic
acid, arachidonic acid and sodium or potassiurn salt
and monoglyceride thereof. These ~atty acid compounds
7Z~
1 may be used alone or as a mixture of two or more kinds
thereof in such a minor amount of 0.004 to 0.1 % (W/V),
and preferably about 0.02 to 0.04 ~o (W/V). Among these
fa.tty acid compounds the preferable ones are those having
14 to 20 carbon atoms and their physiological.ly acceptable
salts, and the most preferable are potassium palmitate
and potassium olea-te, taking into consideration of their
good solubility and ease of the preparation of the emulsion.
The fluorocarbon compound emulsion of this
invention is prepared by homogeneously mixing prescribed
amounts of the aforesaid components in any order in a
physiologically acceptable aqueous medium, such as
distilled water 9 or isotonic solution to obtain a crude
emulsion, and then emulsifying ~he crude emulsion by
injecting it at a temperature of up to 55C through a
slit under a pressure of about 100 kg/cm2 to 500 kg/cm2
thereby subjecting it to shearing force and mixing action
based on a strong velocity gradient, until the desired
.particle size previously mentioned is obtained
The homogeneously mixing of the materials
used is carried out by the use of a conventional mixer
such as homoblender or propeller stirrerO
The emulsification of the crude emulsion is
attained by means of a high pressure homogenizer, which
is a high pressure pump which homogenizes a mixture of
two immiscible liquids by injecting through a slit under
a high pressure at a very high velocity to give a shear
and mixing to the liquids. The typical homogenizer on
market is Manton-Gaulin type homogenizer (Trldema:rk
of this type of homogenizer sold by Manton-Gaulin
.
~7~fZ~
1 Manufacturing Co., Inc., U.S.A.) which has a multiple-
stage valve in combination of -two or more valves each
having a spring therein by which the slits are formed.
The mlxture is circulated in this type of
homogenizer several times under the total pressure of
about 500 kg/cm2 thereby to obtain the stable emulsion
of the invention. The operating temperature is kept
in a range of up to 55C, and preferably 25 to 40C.
The present emulsion has a dispersed phase
of ultrafine par-ticles whose diameter is less than
0.2~4 or at most less than 0.3 ~ . Moreover, it is
stable, showing no growth in particle size, even when
heated or stored for a long period of time. Therefore,
the present emulsion thus secures the administered
animal to a high degree against harmful effect due to
agglomeration of the emulsion particles.
The present emulsion, moreover, has a long
retention time in the circulating blood stream, so
that the oxygen-carrying capacity is maintained for a
long period.
For instance 7 as compared with a fluorocarbon
compound emulsion prepared by use of phospholipids as
emulsifier according to Japanese Patent Application
Kokai (~aid Open) No. 69219/75, the present emulsion
retains for much longer time in the blood stream of
animal. The excretion of the present emulsion from
the body is far faster than that of a perfluorotri-
bu-tylamine emulsion.
The present emulsion may be used as infusion
fluid, after having been made physiologically isotonic.
~(:77;~4~6
1 It may be used also as mixtures with commercial plasma
expanders such as Dextran, hydroxyethylstarch, and
modified gelatin. ~urther, it can be used as a blood
substitute for mammals and as a perfusate for preserving
internal organs.
The present invention is further illustrated
by the following Examples which should not be construed
to limit the invention thereto.
In ~xamples the particle size was measured
by the centrifugal sedimentation method proposed by
K. Yokoyama, A. Suzuki, I. Utsumi and R. ~aito.
~hem. Pharm. ~ull. 22 (12), 2966 - 2971 (1974)~
Example 1
In 8 liters of distilled water, was dissolved
300 g of a polyoxyethylene polyoxypropylene copolymer
Q~ lO~ ~o~
(molecular weight: 8,35~). To the solution were added
40 g of soybean phospholipids, 2 g of potassium oleate,
and a mixture comprising 3 kg of perfluorodecalin and
300 g of perfluorotripropylamine. The resulting mixture
was stirred in a mixer to form a crude emulsion. The
resulting crude emulsion was charged into the liquor
tank of a jet emulsifier (made by Manton-Gaulin Co.)
and emulsified by passing it twelve times through a
valve at a high pressure of 200 to 500 kg/cm2, while
maintaining the liquor temperature at 35 ~ 5C, to
effect emulsification. The resulting emulsion contained
30,5 ~0 (W/V) of perfluorodecalin and 2.9 ~0 (W/V) of
perfluorotripropylamine. The average particle diameter
was 0.09 to 0.1 ~, as measured by the centrifugal
- 10 - `,
''.' ~ :
Z~a4~;
1 sedlmentation method. The emulsion showed substantially
no growth in particle size, when enclosed in a vial for
injection and subjected to thermal sterilization at 115C
for 12 minutes in the special designed rotary sterilizer.
In Table 1 are shown the particle size dis-tribution of
this emulsion and -that of an emulsion of perfluorodecalin
alone prepared without using perfluorotripropylamine.
As seen from Table 1, when stored at 4C for
6 months, the present emulsion did not show any
agglomeration, the mean particle diameter having been
substantially unchanged.
The above procedure was repeated, excep-t
that perfluoropentyltetrahydrofuran was used in place
of the perfluorodecalin, and similar results to those
shown above were obtained.
Example 2
In 8 liters of distilled water, was dissolved
330 g of a polyoxyethylene octyl ether (average molecular
weight: 3,500). To the solution were added 40 g of
soybean phospholipid and 2 g of potassium oleate, and
the resulting mixture was stirred in a mixer to prepare
a dispersion. To the dispersion was added a mixture
comprising 3 kg of perfluoromethyldecalin and 600 g
of perfluoro-~-pentylpiperidine and the resulting
mixture was stirred in a mixer to prepare a crude
emulsion. The crude emulsion was thoroughly emulsified
in the same manner as in Example 1, and the resulting
emulsion was filled in small vials~ The vial containing
the emulsion was subjected to thermal steriliæation at
1Ci~7~
1 115C for 12 minutes in the rotary sterilizer. The
emulsion contained 29.7 ~ (W/V) of perfluoromethyl-
decalin and 5.8 % (W/V) of perfluoro-N-pentylpiperidine.
The particle size distribution after sterilization and
the average particle diameter after storage at 4C ~or
6 months, as well as those of` a reference emulsion of
perfluorodecalin alone were as shown in Table 1.
Example 3
In 2 liters of distilled water, was dissolved
100 g of a polyoxyethylene-polyoxypropylene having an
A~ average molecular weight of ~84~. To the resulting
solution were added 20 g of yol~ phospholipids and
0.5 g of oleic acid, and the mixture was stirred in
a mixer to prepare a dispersion. To the dispersion was
added a mixture comprising 6~0 g of perfluorodecalin
and 250 g of perfluorodibutylmonomethylamine, and the
resulting mixture was stirred in a mixer to obtain a
crude emulsion. The crude emulsion was emulsified in
the same manner as in Example 1 and sterilized by heating
at 115C for 12 min. in the rotary sterilizer. The
emulsion contained 25.3 % (W/V) of perfluorodecalin
and 9.8 % (W/V) of perfluorodibutylmonomethylamine.
The avsrage particle diameter and -the particle size
distribution of the present emulsion and those of a
reference sample of an emulsion prepared by use of
perfluorodecalin alone were as shown in Table 1. In
Table 1 is also shown the average particle diameter
of the present emulsion after storage at ~C f`or 6
months.
~72~
1 ~xample 4
In 800 ml of distilled water, was dissolved
35 g of a polyoxyethylene-polyoxypropylene copolymer
havi.ng an averag~e molecular weight of 15,800. To the
solution were added 4 g of yolk phospholipids and
0.1 g of monoglyceride of lauric acid, and the resulting
mixture was stirred in a mixer to prepare a dispersion.
To the dispersion was added a mixture comprising 350 g
of perfluorohexyltetrahydropyran and 40 g of per1uoro-
N,N-diethylcyclohexylamine, and the resulting mixture
was stirred in a mixer to prepare a crude emulsion.
The crude emulsion was further emulsified in the same
manner as in Example 1, and the resulting emulsion was
subdivided into small portions which were enclosed in
vials. The vial containing the emulsion was subjected
to thermal sterilization at 115C for 12 min. in a rotary
sterilizer. The emulsion contained 35.7 ~ (W/V) of
perfluorohexyltetrahydropyran and 4.1 % (W/V) of
perfluoro-N,N-diethylcyclohexylamine.
The average particle diameter of the present
emulsion and that of a reference sample of emulsion
prepared by use of perfluorohyxyltetrahydropyran alone
after sterilization is shown in Table 1. The present
emulsion showed no change of particle size after storage
at 4C for 6 months.
- 13 -
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- 16 -
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r~ O ~r-l O ~dr-l O ~ r I O
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u~ ~u~ ~ ~ o u~ ~ o
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- 17 -
~7;~4~
1 Experimental Example 1
Test for mixing with plasma expander
For the clinical use as infusion fluid, the
present emulsion is used preferably in combination with
a plasma expander to replenish the defficiency in
oncotic pressure. When the present emulsion was mixed
with a plasma expander, the reversible precipitation
which migh-t be caused by interaction between both
colloidal solutions was not detected, indicating that
one of the difficulties which might be encountered
in using the present emulsion as an infusion fluid
has been eliminated.
The emulsions used in the experiment were
perfluorodecalin-perfluoro-~,~-dibutylmethylamine ;~
(5 : 2) emulsions of various concen-trations prepared
in the same manner as in Example 3 [polyoxyethylene-
polyoxypropylene copolymer, 3.4 % (W/V); yolk phos-
pholipids, 0.6 % (W/V); potassium oleate, 0.04 ~0 (W/V)]
and, as a reference, perfluorodecalin emulsions of
various concentrations prepared according to Japanese
Patent Application Kokai (Laid Open) ~o. 69219/75
[yolk phospholipid, 4 % (W/V); potassium oleate,
0.02 % (W/V)]. Each emulsion was made isotonic with
the lactated Ringer's solution or Krebs-Ringer bi-
carbonate solution, then admixed with a plasma expanderso that the final concentration of the latter may
become 1 to 6 ~ (W/V), and forma-tion of precipitates
was visually observed during 6 hrs. after mixing at
room temperature. Plasma expanders used are hydroxy-
~0 ethyistarch (HE~) (average molecular weight: 200,000
- 18 -
~7;~44~
1 20 ~ (W/V) in saline, supplied by Ajinomoto Co., IJtd.)
and Dextran 40 [Dextran (average molecular weight,
40,000) 10 ~ (W/V) in saline in water; supplied by
The Green Cross Corp.]~
The results are shown in Tables 2 and 3.
Table 2
_
\ Final fluoro- Emulsion, Jap.
\ carbon Emulsion of the Pat. Appl. ~aid
\ conten-t, invention Open No.
Einal \ ~0 (W/V) 69219/ 75
Dextran 40 con~
Table
\ Final fluoro- _ ~ Emulsion, Jap.
\ carbon content Emulsion of the Pat. Appl. I,aid
\ % (W/V) invention Open No.
Final \ 69219/75
~ 1~
~ 19 -
~ ~ 7 ~
1 Note: -: non precipita-tes
+: formation of precipitates
Form the results obtained above, it was made
clear that the p-resent emulsion is much less affected
by the presence of a plasma expander compared wlth
the emulsion according to Japanese Patent Application Kokai
(~aid Open) ~o. 69219/75, indicating that -the present
emulsion can be mixed wlth the Dextran 40 and HES
porf/on
preparation in any~ to make it the physiologically ~: .
colloidal isotonicity which obtained by addition
of Dextran 40 and HES as a final concentration of
2 % (W/V) and 3 ~0 (W/V), respectively.
Similar results to those mentioned above were
also found in the emulsions prepared in Examples 1,
2, and 4.
Experimental ~xample 2
In order to evaluate the efficacy of the
present emulsion, the study on the exchange-transfusion
in rats was done.
- 20 -
~72~
o _
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~1 ~ 0~ ~D O O ~ ) O r I
~ ~ N ~ O O~O N O I O O ~i0~ O
_ _ _ ~ '
* -~ ~ ~
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.~ O O P~ ~Q~ ~ \I (I) Q~
~ C)O ~ ~i r-l
E-l O ~ +~ ~ ~, r~ ~ ~ ~ ~1) N O O
r I r I ~¦ r~ U~ O
tH 4-1 05 l ~I r~l tl) r-l V ~r l r-l r-l r-l r~
~I h ~ :5r~ O+~V ~:q r-l ~ V V~ID O
a.) O O r~O ,!~ O ~ ~ V bD C~ I ~4
P 1 ~4 ~ 1~4 ~ ~ P I ~; ~ ~ U2 r~l V F~ a)
1:> _ _ . .~
r-
~r l h
r~ ~1 ~I td ~q
H ¢l ~V~ +) ~ ~ X
O ~ r~
,r~ ^ r~ *
o r~ ~r-l r-l r~
O ,D r ~
- 21 -
~C~7~4~1~
1 Two kinds of fluorocarbon compound emulsion
which were the present emulsion, the mixture of per-
fluorodecalin and per-fluorodibutylmonome-thylamine
prepared in Example 3 and per~luorodecalin emulsion
stabilized with yolk phospholipids according to
Japanese Patent Application Kokai (Laid Open) No.
69219/75 were used in this experiment.
The ingredients of both emulsion were shown
in Table 4.
To make the emulsion electrolyte and colloidal
isotonification, one volume of hypertonic electrolytes
solution shown Table 4 is added to 9 volume of the
emulsion, and then9 1 volume of resultant emulsion
containing electrolytes were mixed with 3 volume
f 6 % hydroxyethyl starch (molecular weight 40 000 -
50,000) in Ringer's lactated solution or rat plasma
as reference, prior to use.
sJ~er
The rats (~ri~ strain weighing 200 to
250 g) were exchange transfused with the emulsion
containing electrolytes and hydroxyethyl starch or
plasma by repeated bleeding from carotid artery and
replacement transfusion through tail vein alternately
up to hematocrit 1 %, 4 % and 7 %, respectively~ under
the 100 % oxygen atomosphere.
Then, the survival time of rats exchange~
transfused were determined.
The results are shown in Table 5. As is
evident in Table 5, the present emulsion was far more
effective in saving the life o-f animal hemorraged
massively in comparing with the perfluorodecalin
- 22 -
~7~
1 emulsion st~bilized ~lith yolk phospholipids.
Table 5
. Jointly withJointly with
Flnal plasma ~ES
hematocrit _ _
value % ~urvival time
7> 72 hours> 72 hours
The present 4 50 0O min. ~ 72
emulsion
1 29 02 61 oo min.
_
~mulsion 7 23 hUrs 10 min. 5 hours 53 min-
acc. to Jap.
Pat. Appl. 4 11 5 5 2
~aid Open
~o. 69219/75 1 8 10 2 56
. _
~ote: The values obtained from 5 rats in each
group.
~xperimental Example 3
The present fluorocarbon compound emulsion
obtained in Example 3 and the emulsion obtained accord-
ing to Japanese Patent Application Kokai (~aid Open)
No. 69219/75 were subjected to acute toxicity test.
In order -to make these emulsion isotonifica-
tion, one volume of electrolytes is added to 9 volume
f the emulsion prior to use The ingredients of these
emulsions were shown in Table 4. As test animals were
used wister strain male rats weighing 100 to 120 g.
The rats were intravenously injected with the emulsion
and the survival rate was observed during one week
after injection. The results obtained are shown in
4~
1 Table 6. As seen in Table 6, the ~Dso of both emul-
sions was around 130 ml/kg body weight, indicating
they were quite low-toxic.
- 2l~ -
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o
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a) u~
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OS~ td ~ LS~
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ha) r~ a) ~1
C~~ ~ ~ ~r~
c~~ ~ a
O~ rJ ~ ~
hP u~ O O O O O
Or~ O ~ ~1 -1 ~1 ~1~1
::~~d ~ ~ a
P ~ ~ o ~ o~ ~D
o o ~ ~
u~ ~ a) ~ o o o o o
a) ~ ~ ~1 ~1 ~ ~1~1
~H~' F~ ~ N
P~~ ~ ~ l ~ ~
rl
C) O O O O O
.
~C ~1
~0 ~0 ~ ~ ~ ~
$ _ _
c~ ~D
c~ o ~ 1~ O u~
~o o~ c~ ~ ~ ~ ~1 '
~ :
H
a
a) ta ~
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E3 h ~rl
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- 25 -
~72
__ __
o ~
o a~ ~ Lf~ ~
o ~ o o o
~ o~ ~ ~ ~
._ o o o o o
~) ~ a~ c~ ~ ~
o o o o o
~ ~ ~ ~ ~1
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~D O O O O O
a) ~ ~1
o o o
V
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0 ~o ~ ~ ~
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C~ o U~
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~ O ~,o
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- 26 --
~L~7Z4~Lb;
1 ~xperimental ~xample 4
In order to examine hemolytic effect of the
fluorocarbon preparations in the c~tracorporea:L circula-
tion system, experiments in vitro were carried out by
using red blood cells of rabbit.
Two emulsion preparations used in Experi-
mental Example 2 as shown in Table 4 were admixed
with a lactated Ringer's solution so as to become
substantially isotonic physiologically. The result-
ing isotonic emulsion preparations were mixed withthe heperinized rabbit blood in a ratio of 3 : 1,
1 : 1, and 1 : 3 to prepare sample solutions for the
tests. The hemolytic effect was evaluated by measuring
the free hemoglobin content of 8 ml of the blood after
having been kept at 37C for 6 hours. Determination
of hemolyzed hemoglobin was determined by the cyanomethemo-
globin method (Kampen, ~. J. and Ziilstram, W. J.
Clin. Chim. Acta. 6, 538, 1961).
The results obtained are shown in Table 7.
Table 7
______ ~ _ ~ _
Free hemoglobin, mg
. _ . _
Fluorocarbon : blood ratio 1 : 3 1 : 1 3 : 1
Fmulsion of this invention 36 81 180
Emulsion acc. to Jap. Pat.
Appl. ~aid Open No. 298.5> 4,000 3,380
69219/75
~actated Ringer's solution 36 128 141
(r~r~ ' _. _
~7~4~
1 As seen from Table 7, the hemolytic effect
of the present emulsion is far smaller than that of
prior art and not much different from that of the
lactated Ringer's solution used as reference.
- 28
