JPH0521090B2 - - Google Patents

Description

[Detailed description of the invention]

(Industrial Application Field) The present invention relates to a hemolysis inhibitor. Specifically, the present invention relates to a hemolysis inhibitor that has extremely high safety and is capable of extremely effectively suppressing the hemolytic phenomenon that occurs during blood storage. (Prior Art) When a solution containing red blood cells, such as whole blood or a concentrated red blood cell (CRC), is stored for a long period of time, a so-called hemolysis phenomenon occurs in which the red blood cell membrane is destroyed and hemoglobin is released to the outside world. The main factors that cause hemolysis include changes in osmotic pressure caused by differences in ionic composition in the blood, differences in colloid osmotic pressure due to protein components such as hemoglobin, changes in membrane proteins and lipids of red blood cells, and changes in Na ⁺ and K ⁺ failure of active transport,
Examples include the effects of drugs and poisons. By the way, it has recently been revealed that di-2-ethylhexyl phthalate (DOP), which is commonly used as a plasticizer for vinyl chloride resin, has an effect of suppressing hemolysis (Blood 64 6 1270-(1984) [Blood
64 6 1270-(1984)]). However, this
When blood is stored in a storage container made of di-2-ethylhexyl phthalate plasticized vinyl chloride resin, which is conventionally used as a medical vinyl chloride resin composition, the di-2-ethylhexyl phthalate eluted into the blood reduces platelet aggregation ability. was reported to be suppressed (Journal of the Japanese Society of Blood Transfusion, 28(3)282).
(1981)), it is undesirable for di-2-ethylhexyl phthalate to enter the body together with stored blood during blood transfusion because of its effect on platelets. When I replaced it with a material that does not contain ethylhexyl phthalate,
This was discovered because hemolysis of red blood cells occurred significantly during storage. Therefore, as a countermeasure to this phenomenon, di-2-ethylhexyl phthalate is added to blood stored in a storage container made of a material that does not elute plasticizers (or does not contain plasticizers) to suppress hemolysis. Although contradictory methods have been proposed (US Pat. No. 6,326,025),
The use of 2-ethylhexyl phthalate as an antihemolytic agent was not desirable from the viewpoint of physiological safety. (Means for Solving the Problems) Therefore, an object of the present invention is to provide a novel hemolysis inhibitor. Another object of the present invention is to provide a hemolysis inhibitor that has excellent preservation properties for red blood cells and excellent physiological safety. A further object of the present invention is to provide a hemolysis inhibitor that can be incorporated into a resin composition or directly added to a liquid containing red blood cells. (Means for Solving the Problems) The above objects are achieved by a hemolysis inhibitor comprising an unsaturated chain hydrocarbon compound having at least one double bond and having 8 to 40 carbon atoms. . The present invention also provides an antihemolysis agent, wherein the unsaturated chain hydrocarbon compound is selected from the group consisting of 7-tetradecene, 8-octadecene, 9-eicosene and squalene. The present invention further provides a hemolysis inhibitor that is incorporated into the synthetic resin composition. The present invention also provides an antihemolytic agent that is added directly into the red blood cell containing solution. The invention further provides an anti-hemolysis agent which is added to the red blood cell-containing solution in the form of an emulsion. (Function) The hemolysis inhibitor of the present invention is characterized by being composed of an unsaturated chain hydrocarbon compound having at least one double bond and having 8 or more and 40 or less carbon atoms. Surprisingly, the above-mentioned unsaturated chain hydrocarbon compounds have the same effect of preventing hemolysis of red blood cells as di-2-ethylhexyl phthalate, and on the other hand, unlike di-2-ethylhexyl phthalate, they have no platelet aggregation ability. It was found that no inhibitory effect was observed. Furthermore, the unsaturated chain hydrocarbon compound described above can be dispersed in a solution containing red blood cells, and in particular, if it is in the form of an emulsion, it can be dispersed more uniformly, so it can also be added directly to a solution containing red blood cells. on the other hand,
The unsaturated chain hydrocarbon compounds mentioned above have sufficient compatibility with various synthetic resins including vinyl chloride resins, so the unsaturated chain hydrocarbon compounds can be used in synthetic resin compositions. The action of the hemolytic inhibitor can also be exerted by incorporating it into the hemolytic agent. That is, when a synthetic resin composition containing the unsaturated chain hydrocarbon compound comes into contact with a solution containing red blood cells, the unsaturated chain hydrocarbon compound is eluted and transferred from the resin composition to the solution containing red blood cells. Furthermore, the same effect as when directly added to a red blood cell-containing solution can be obtained. Hereinafter, the present invention will be explained in more detail based on embodiments. The hemolytic inhibitor of the present invention is composed of an unsaturated chain hydrocarbon compound having at least one double bond and having 8 or more and 40 or less carbon atoms. In the above-mentioned unsaturated chain hydrocarbon compound, the number and position of double bonds are not particularly limited, and the presence of at least one double bond can be expected to prevent hemolysis. Also, increase the number of carbon atoms to 8 or more and 40
If the number of carbon atoms is less than 8, it may cause hemolysis, while if the number of carbon atoms is more than 40, unsaturated chain hydrocarbon compounds may be used in whole blood, concentrated red blood cells, etc. This is because it becomes solid in the storage temperature range of red blood cell-containing solutions, making it difficult to disperse well in red blood cell-containing solutions, making it impossible to expect hemolysis prevention effects. Specifically, the unsaturated chain hydrocarbon compounds include octenes, nonenes, decenes, undecenes, dodecenes, tridecenes, tetradecenes, pentadecenes, hexadecenes, heptadecenes, octadecenes, and nonadecenes. , eicosene, octadecadiene, octadecatriene, eicosatetraene, ocimene, myrcene, squalene and the like, with 7-tetradecene, 8-octadecene, 9-eicosene and squalene being particularly preferred. Hemolytic inhibitors made of such unsaturated chain hydrocarbon compounds are used by being directly added to solutions containing red blood cells, such as whole blood and concentrated red blood cells.
dextrose) solution and CPD (citrate phosphate)
(including cases in which it is stored in advance with an anticoagulant/preservative solution such as dextrose solution and added to a solution containing red blood cells), or it can be placed in a blood storage container such as a blood bag, a catheter, a blood transfusion set, a blood circuit, etc. It can also be used by being blended into a medical melting synthetic resin composition constituting a medical device that comes into contact with a red blood cell-containing solution. When a hemolysis inhibitor made of an unsaturated chain hydrocarbon compound as described above is added directly to a solution containing red blood cells, the unsaturated chain hydrocarbon compound may be added as is, but it is preferable that the unsaturated chain hydrocarbon compound be added directly to the solution containing red blood cells. It is desirable to add it in the form of an emulsion so that uniform dispersion and mixing can be achieved. To prepare the emulsion of the unsaturated hydrocarbon compound, a surfactant that does not have an adverse effect on blood components, such as polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene solvitamin monopalmitate ( Tween40), polyoxyethylene sorbitan monostearate (Tween60), polyoxyethylene sorbitan monoesters (Tween series, etc.) such as polyoxyethylene sorbitan monooleate (Tween80), polyoxyethylene sorbitan monoesters (Tween series, etc.), polyoxyethylene pyrioxypropylene block copolymers (Pluradot HA-
430 BASF, etc.), sorbitan monoacyl laurate (Span20), sorbitan monoacyl palmitate (Span40), sorbitan monoacyl stearate (Span60), sorbitan monoacyl oleate (Span80) series, etc.) or hydrogenated castor oil, etc., and dispersed in an appropriate aqueous medium such as physiological saline or buffer solution, or in an anticoagulant/preservative solution such as ACD solution or CPD solution. . The amount added when directly added to a red blood cell-containing solution will vary depending on the red blood cell-containing solution and the type of unsaturated chain hydrocarbon compound, but for example, for whole blood, the final concentration is
100μM to 10mM, more preferably 400μM to 4m
It is desirable that it is M. On the other hand, when the hemolysis preventive agent made of the unsaturated chain hydrocarbon compound is blended into a medical synthetic resin composition, the unsaturated chain hydrocarbon compound is, for example, vinyl chloride resin, polyolefin resin, polystyrene resin, etc. It is possible to uniformly disperse and mix the unsaturated chain hydrocarbon compound into various resin compositions such as polyester resins, poly(meth)acrylic resins, and polycarbonate resins. It may be added to the synthetic resin composition. The synthetic resin composition blended with the unsaturated chain hydrocarbon compound in this way can be formed by any conventional molding method, such as calendar molding, extrusion molding, injection molding, plastisol molding, etc. It can be molded, and conventionally used high frequency fusion and heat fusion can be used as an adhesion method. When the above-mentioned unsaturated chain hydrocarbon compound is blended into such various synthetic resin compositions, the unsaturated chain hydrocarbon compound eluted and transferred from the synthetic resin composition comes into contact with the red blood cell-containing solution and dissolves the red blood cells. Since it provides a protective effect, its amount varies depending on the synthetic resin composition and the type of unsaturated chain hydrocarbon compound, but it is generally 1 to 20% by weight in the resin composition. , more preferably 3 to 10
It is said to be about % by weight. If the blending amount of the unsaturated chain hydrocarbon compound is within this range, an effective red blood cell protection effect can be exerted when a solution containing red blood cells comes into contact with a medical device formed from the synthetic resin composition. In addition, the physical properties of the synthetic resin composition are not substantially deteriorated. In addition, in an embodiment in which a hemolysis preventive agent made of such an unsaturated chain hydrocarbon compound is blended into a medical synthetic resin composition, a particularly preferred example is when the medical synthetic resin composition is a soft vinyl chloride resin. For example, it is a composition. In other words, conventionally, soft vinyl chloride resin compositions constituting medical devices that come into contact with red blood cell-containing solutions, such as blood bags, have been used as plasticizers, even though it is known that they have a platelet aggregation inhibiting effect to prevent hemolysis. −
2-ethylhexyl phthalate was used, but if the hemolysis preventive agent according to the present invention is blended, it is possible to use a plasticizer with higher safety and more preferably low elution properties, such as di-n-octyl phthalate and di-n-octyl phthalate. di-normal alkyl phthalates such as normal nonyl phthalate, di-normal decyl phthalate, di-normal undecyl phthalate, and dilauryl phthalate; trialkyl trimellitates such as tri-2-ethylhexyl trimellitate and tri-normal octyl trimellitate; Tetraalkylpyromellitates such as tetra-2-ethylhexylpyromellitate can be used, and if such plasticizers are used, di-
Unlike when 2-ethylhexyl phthalate is used as a plasticizer, there is no adverse effect such as inhibition of platelet aggregation due to elution and transfer of plasticizer.
On the other hand, since the hemolysis preventing effect can be brought about by the hemolysis preventing agent according to the present invention, it is possible to obtain an extremely excellent medical soft vinyl chloride resin composition. (Examples) Hereinafter, the present invention will be described in more detail based on Examples. Example 1 2000μ of 7-tetradecene (manufactured by Tokyo Kasei Co., Ltd.)
g/ml polyoxyethylene monooleate (Tween 80, manufactured by Wako Pure Chemical Industries, Ltd., special grade)/physiological saline to a concentration of 10 mM. Add 0.2 ml of this emulsion to 1.8 ml of human CPD red blood cell concentrate (hereinafter referred to as CRC) adjusted to a hematocrit value of approximately 70% to give the final concentration shown in Table 1, and place in a polypropylene tube with a stopper. It was stored for 4 weeks at 4°C. Thereafter, the plasma hemoglobin concentration was measured using the TMB method (Clinical Chemistry 23 749~ (1977) [Clin,
Chem. 23 749-(1977)]). The results are shown in Table 1. Example 2 An emulsion was prepared in the same manner as in Example 1, except that 7-tetradecene was dispersed in a polyoxyethylene monooleate/physiological saline mixture to a concentration of 4 mM, and it was added to a CRC and allowed to stand still. Plasma hemoglobin concentration was measured after storage. The results are shown in Table 1. Example 3 An emulsion was prepared in the same manner as in Example 1, except that 8-octadecene was dispersed in the polyoxyethylene monooleate/physiological saline mixture at a concentration of 20 mM instead of 7-tetradecene.
It was added to CRC and the plasma hemoglobin concentration was measured after static storage. The results are shown in Table 1. Examples 4-5 Same as Example 3 except that 8-octadecene was dispersed in the polyoxyethylene monooleate/saline mixture to a concentration of 10mM (Practical Example 4) or 4mM (Example 5). An emulsion was prepared in the same manner, added to CRC, and the plasma hemoglobin concentration was measured after static storage. The results are shown in Table 1. Example 6 Squalene (Sigma
S3626) as polyoxyethylene monooleate/
An emulsion was prepared in the same manner as in Example 1 except that the emulsion was dispersed in a physiological saline mixture to a concentration of 20 mM, and the emulsion was added to CRC and the plasma hemoglobin concentration was measured after static storage. The results are shown in Table 1. Examples 7-8 Same as Example 6 except that squalene was dispersed in the polyoxyethylene monooleate/physiological saline mixture to a concentration of 10mM (Practical Example 7) or 4mM (Example 8). An emulsion was prepared, added to CRC, and the plasma hemoglobin concentration was measured after static storage. The results are shown in Table 1. Comparative Example 1 An emulsion was prepared in the same manner as in Example 1, except that di-2-ethylhexyl phthalate was dispersed in the polyoxyethylene monooleate/physiological saline mixture to a concentration of 5 mM instead of 7-tetradecene. It was prepared, added to CRC, and the plasma hemoglobin concentration was measured after static storage. The results are shown in Table 1. Comparative Example 2 Only a 2000 μg/ml polyoxyethylene monooleate (Tween 80, manufactured by Wako Pure Chemical Industries, Ltd., special grade)/physiological saline mixture was added to the CRC, and the plasma hemoglobin concentration was measured after static storage. . Results first
Shown in the table.

[Table] Examples 9-10 and Comparative Examples 3-4 After rolling-kneading the soft vinyl chloride resin compositions having the compositions shown in Table 2 at 160°C for 10 minutes,
It was molded into a 0.4 mm thick sheet using an extruder. A 20 ml mini-blood bag was created by overlapping two of the obtained sheets and applying high-frequency sealing to predetermined areas. Approximately 20 ml of CRC adjusted to a hematocrit value of approximately 70% was dispensed into the bag and stored at 4°C for 4 weeks. Thereafter, plasma hemoglobin concentration was measured in the same manner as in Example 1. The results are shown in Table 3.

[Table] Table 3 Plasma hemoglobin concentration [mg/dl] Example 9 67 Example 10 58 Comparative example 3 46 Comparative example 4 91 As is clear from the results shown in Tables 1 and 3, hemolysis prevention of the present invention The plasma hemoglobin concentration values of CRC (Examples 1 to 10) to which the agent unsaturated chain hydrocarbon compound was added in any form were as follows:
This value was almost close to the plasma hemoglobin concentration of CRCs to which di-2-ethylhexyl phthalate was added in any form (Comparative Examples 1 and 3), indicating a good hemolysis prevention effect. Examples 11-13 and Comparative Examples 5-6 40mM methanol solution of 7-tetradecene (Example 11), 40mM methanol solution of 8-octadecene (Example 12), 40mM methanol solution of squalene (Example 13), di- 4mM methanol solution of 2-ethylhexyl phthalate (Comparative Example 5) or methanol without any addition (Comparative Example 6)
was added to human platelet-rich plasma (platelet count: approximately 300,000/mm ³ ) in 1/100 amount, and after being left at room temperature for 2 hours, platelet aggregation ability was measured using an Agricorder (manufactured by Kyoto Daiichi Kagaku Co., Ltd.). . The percentage of the light transmittance of each sample-added platelet-rich plasma solution relative to the maximum light transmittance of the platelet-rich plasma solution alone (blank) was expressed as the aggregation rate. Furthermore, 5 μM ADP (adenosine diphosphate) and 5 μg/ml collagen were used as aggregation-inducing substances. The results obtained are shown in Table 4.

[Table] As shown in Table 4, di-2-ethylhexyl phthalate has an inhibitory effect on recovery of platelet aggregation ability. This suggests that di-2-ethylhexyl phthalate may inhibit platelet function when it enters the body, indicating that it is not suitable as a hemolysis inhibitor. On the other hand, 7-tetradecene, 8-octadecene, and squalene, which are unsaturated chain hydrocarbon compounds related to the hemolysis inhibitor of the present invention, have no such inhibitory effect and are considered to be safer substances. This indicates that the hemolysis inhibitor of the present invention has little effect on platelets. (Effects of the Invention) As described above, the present invention is a hemolysis inhibitor made of an unsaturated chain hydrocarbon compound having at least one double bond and having 8 to 40 carbon atoms. It has an extremely high level of safety, as it has low oxidation and does not inhibit platelet aggregation ability, and extremely effectively suppresses the hemolysis phenomenon that occurs during storage of red blood cell-containing solutions such as whole blood or red blood cell concentrate (CRC). Furthermore, the hemolysis inhibitor of the present invention can be added directly to the red blood cell-containing solution, preferably in the form of an emulsion, or can be added directly to the synthetic resin constituting the medical device with which the red blood cell-containing solution comes into contact. It can be used in a wide range of situations, including being able to be incorporated into compositions, and can be said to be an extremely excellent hemolysis inhibitor. Further, in the hemolysis preventing agent of the present invention, further effects can be expected when the unsaturated chain hydrocarbon compound is selected from the group consisting of 7-tetradecene, 8-octadecene, 9-eicosene, and squalene. becomes.

Claims (1)

[Claims] 1 Having at least one double bond and having 8 carbon atoms
A hemolysis inhibitor comprising an unsaturated chain hydrocarbon compound having a molecular weight of 40 or more. 2. The hemolysis inhibitor according to claim 1, wherein the unsaturated chain hydrocarbon compound is selected from the group consisting of 7-tetradecene, 8-octadecene, 9-eicosene, and squalene. 3. The hemolysis inhibitor according to claim 1 or 2, which is blended into a synthetic resin composition. 4. The hemolysis inhibitor according to any one of claims 1 to 3, which is added directly to a red blood cell-containing solution. 5. The hemolysis inhibitor according to claim 4, which is added to the red blood cell-containing solution in the form of an emulsion.