JPH037166A - Extra-body circulating circuit - Google Patents

Abstract

PURPOSE:To treat malignant tumor simply, safely, and continuously by equipping a blood plasma processing means with a mixing machine for promotion of the contact with the plasma processing liquid, and therewith processing the blood sampled from a living body effectively. CONSTITUTION:An extra-body circulation circuit includes a blood plasma separating means 1, a plasma processing means 200 equipped with mixing machine, an adjusting means 300, a mixer 4 for the processed plasma and blood corpuscle components, and a circulation means 600 consisting of pumps 61-63 and tubes 71-75. The blood taken from an artery of a living body of for ex. house rabbit 5 is delivered to the plasma separating means 1 by the circulation means 600 for ex. pump 61, and part of the plasma is segregated. The blood components having passed through this segregation is returned back to the living body by a vein 54, etc. The segregated plasma components are fed to the plasma processing means 200 and put in contact with the plasma processing liquid. It is then adjusted for coordination with the intra-body environment by the adjusting means 300, circulated back by the circulation means 600 for ex. pump 63, and mixed in the mixer 4 with the blood components not segregated, and this mixture is returned into the body.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to an extracorporeal circulation circuit used for treatment of malignant tumors and the like.

(Prior technology) "Double filtration plasma separation method" as a treatment method for intractable diseases such as malignant tumors, autoimmune diseases, liver failure, and DIC1 hyperlipidemia (edited by Tetsuzo Agishi; Igaku Shoin, 1984) ) has been proposed. In this method, blood is continuously drawn from the patient and separated into plasma and blood cells using a separation membrane.
After the large molecular weight fraction is removed from the obtained plasma using another separation membrane (blood fractions such as albumin fraction are left behind), the blood is returned to the patient together with the blood cell components.

The above-mentioned large molecular weight fractions contained in plasma are thought to be various specific and non-specific immunosuppressive substances present in the blood of cancer-bearing patients, for example. Immunosuppressive substances belonging to such a large molecular weight fraction, for example, form a large matrix by binding various substances such as antigens, antibodies, and complement to antibodies produced by the surface of malignant tumor cells as specific antigens. It is thought to be an immune complex. In patients with malignant tumors, immunosuppressive substances such as these cause a decline in immune function, and as a result of a complex interplay of these substances and other factors, 1ffii cells are no longer able to maintain their normal immune surveillance mechanism. It is said that it deviates and proliferates and metastasizes. Therefore, malignant tumors can be improved by selectively removing large molecular weight fractions as in the above method. However, such methods of removing immunosuppressive substances do not have the effect of actively attacking malignant tumor cells and causing necrosis. Furthermore, when removing large molecular weight fractions using a separation membrane, 1. There is also a possibility that a portion of plasma proteins necessary for the living body may be removed.

In addition, as an example of improvement of malignant tumors by blood treatment, see this other paper [Acute tumor necrosis obtained by intravenous administration of serum from tumor-bearing rabbits treated with hypertonic salt] (Takashi Yamamoto, Ri! Xbed Immunization, June 1986 issue, pages 544-547). According to this report, serum obtained from tumor-bearing rabbits is mixed with a concentrated sodium chloride aqueous solution, the sodium chloride concentration is lowered by dilution or dialysis, and the treated serum is returned by intravenous injection. . It has been confirmed that cancer shrinkage is caused by such treatment.

In addition, the treatment method for this malignant tumor was published in Japanese Patent Application Laid-Open No. 63-9133.
It is also disclosed in Publication No. 0. That is, [(1) contacting the plasma or serum obtained from a living body with a malignant tumor with a hypertonic salt solution, and (2) contacting the plasma or serum obtained in the step (1) above with the same or A method for treating a malignant tumor, comprising the step of administering it to a living body of the same species. It is J. The mechanism of action is believed to be that the immune complex is denatured or dissociated and the activity of the antitumor antibody is restored. Furthermore, this inventor has developed a device for use in this treatment in Japanese Patent Application Laid-Open No. 63-102763.
It is disclosed in the publication No. However, this device does not provide a means to promote contact between plasma and hypertonic salt solution, and therefore does not efficiently mix plasma and hypertonic salt solution, which have significantly different specific gravities. Therefore, sufficient therapeutic effects may not be obtained.

In addition, as a treatment method for such malignant tumors,
3-93730 discloses that as a method for treating plasma, it is effective to treat it not only with hypertonic salts but also with acidic solutions.

As an extracorporeal circulation circuit for this treatment, JP-A-63-
No. 200768 is disclosed, but there is no disclosure of means for efficiently mixing plasma and processing liquid.
In this case as well, a sufficient therapeutic effect may not be obtained.

(Problems to be Solved by the Invention) The present invention solves the above-mentioned conventional drawbacks, and its purpose is to provide a system that can effectively treat malignant tumors, etc. without removing active biological components; It is an object of the present invention to provide the above-mentioned system which can treat malignant tumors etc. in a simple, safe and continuous manner by efficiently processing blood collected from a living body.

(Means for Solving the Problems) The extracorporeal circulation circuit of the present invention includes a plasma separation means for separating plasma from blood collected from a living body, and a plasma separation means for separating plasma from blood collected from a living body, and an inorganic salt solution, an organic salt solution, and an organic salt solution. A plasma treatment means for treating plasma by contacting it with at least one selected from the group of plasma treatment solutions consisting of acidic solutions, a conditioning means for adjusting the treated plasma to the in-vivo environment, and a conditioned plasma. and a reflux means capable of continuously or intermittently refluxing plasma into the living body, the plasma processing means comprising:
The circuit is equipped with a mixer for promoting contact between the plasma and the plasma processing solution, and the above objective is achieved by this circuit.

For example, as shown in FIG. 1, the circuit of the present invention includes a plasma separating means 1, a plasma processing means 200 (encircled by a dotted line) equipped with a mixer, a regulating means 300, and a mixer 4 for mixing treated plasma and blood cell components. , pumps 61, 62゜63-, etc. and tube 7
It has a reflux means 600 consisting of 1, 72, 73, 74, 75, etc. Blood from an artery 51 of a living body such as a domestic rabbit 5 is sent to the plasma separation means 1 by a reflux means 600 such as a pump 61, and a part of the plasma is separated. The unseparated blood components are returned to the living body through a vein 54 or the like.The separated plasma components are supplied to a plasma processing means 200 equipped with a mixer, where they are mixed with blood. After being brought into contact with the treatment liquid and then adjusted to the in-vivo environment by the adjustment means 300, refluxed by the reflux means 600 such as the pump 63, and mixed with unseparated blood components by the mixer 4. , is returned to the body.

The plasma separation means 1 may be of a hollow fiber type or a centrifugal type, and preferably a hollow fiber type.

This plasma processing means 200 includes a plasma processing liquid tank 21, a pump 2
2. It is composed of a processing reaction tank 23, a mixer 24, and a cock 25, and the processing liquid tank 21 stores a plasma processing liquid. The shape of the mixing device 24 is not particularly limited as long as it can be placed outside the plasma processing reaction tank 23 and used. Examples include an overturning rocker platform mixer, a rotary shaker, and an ultrasonic mixer with a cooler. The shape of the container of the plasma processing reaction tank 23 is not particularly limited as long as it can be used with the mixer described above. Examples include a box-shaped reaction tank, a syringe-shaped reaction tank, a bag-shaped reaction tank, and the like.

Plasma processing is performed by this plasma processing means 200, for example, as follows. With the cock 25 closed, plasma and plasma processing liquid are fed into the plasma processing reaction tank 23 by the pump 62 (reflux means) and the pump 22, respectively, and the mixer 24 is started at the same time as the feeding starts, and both are mixed. promotes mixing.

As the plasma treatment liquid, at least one of an inorganic salt solution, an organic salt solution, and an acidic solution is used. Inorganic salts used include sodium chloride, potassium chloride, magnesium chloride, magnesium sulfate, sodium phosphate, potassium phosphate, ammonium phosphate, ammonium sulfate, sodium borate, potassium borate, and the like. Organic salts include, for example, sodium citrate, potassium citrate, sodium acetate, potassium acetate, sodium birophosphate, potassium pyrophosphate, sodium phthalate, potassium phthalate, sodium fumarate, potassium fumarate, sodium tartrate, potassium tartrate. , sodium succinate, potassium succinate, sodium formate, potassium formate, sodium lactate, and potassium lactate. Furthermore, r
PIPES- is known as a component of Good's buffer solution.
Sodium, PrPE5-potassium, MOPS-sodium, MOPS-potassium, HEPES-sodium,
HEPES-potassium, Tris-hydrochloride, glycine-
Hydrochloride, Tricine Hydrochloride, TAPS-Sodium, TAPS-Potassium, CAPS-Sodium, CAP
Examples include S-potassium, TBS-sodium, TES-potassium, and Bicine hydrochloride. The above-mentioned inorganic salts and organic salts have a pH of 6.0 or less depending on their paired acid or base or a combination thereof. Provide a buffering effect in the range of 0 to 8.0, or use an appropriate buffer solution to adjust the pH to 6,0 to 8.0.
These inorganic salts or organic salts are preferably dissolved in an aqueous solution of 0.5M or more, preferably about 1 to 4M (
Hypertonic salts) and plasma 1#11! per 0.1-100m
, preferably at a ratio of 0.5 to 30 tnl.

As the acidic solution, common aqueous inorganic or organic acids such as hydrochloric acid, phosphoric acid, acetic acid, and citric acid can be used, and various acidic buffers are preferably used. Examples of acidic buffers include glycine-hydrochloric acid buffer, citrate buffer, acetate buffer, phosphate buffer, Boron M buffer, potassium phthalate-hydrochloric acid buffer, and the like. The pH of the acidic aqueous solution is adjusted to be 5.0 or less, preferably 3.5 or less (usually 10-3 mol or more) when mixed with plasma.

The adjusting means 300 is, for example, a desalting device or a diluting device, and has a function of adapting the plasma, which has been processed in the plasma processing means 200 and has a high salt concentration or low pH, to the original in-vivo environment. have

As the desalting device, a dialysis device, an ion exchanger, a gel filtration device, etc. are used. For example, a hollow fiber type dialyzer is suitable as the dialysis device.

In addition, to remove NaCJ using an ion exchange resin, a cation exchange resin column excluding Na and an anion exchange resin column excluding CI are sequentially arranged. A liquid feeding path (not shown) is provided to remove the salt eluted from the circuit.

The shape of the mixer 4 for mixing treated plasma and blood cells is not particularly limited, but a common air trap or the like may be used.

Further, the mixer 4 may be simply connected to the blood cell component flow path and the plasma flow path without using a device having a special shape.

In addition, pressure detectors 81, 82, 83 are installed at appropriate locations in the circuit.
An example of these is a billows pressure sensor.

(Example) Below, experimental examples and examples of the present invention will be given and further explained.

Using the apparatus shown in Figure 2 above, an experiment was conducted to confirm the effect of mixing plasma and plasma processing solution in vitro.

As a plasma model, an aqueous bovine albumin solution (-/C = 7%) (hereinafter referred to as BSA solution) whose protein concentration was matched to that in the blood of a living body was used. In FIG. 2, the BSA solution described above is placed in the container 50, a blood bag (Chill Seven Separation Pang Chil Flex 150mj!, Chill Seven Co., Ltd.) is used as the plasma processing reaction tank 23, and the plasma processing liquid tank 21 is A 3M aqueous sodium chloride solution was placed in the container.

Sodium chloride aqueous solution and BSA? The solution was supplied to the reaction vessel 23 at a flow rate of 8 II dl/sin and mixed using a double peristaltic pump 22 (thus, both solutions were supplied at the same rate). For mixing, a rocker platform for DNA extraction (Shiraki Seisakusho) was used as the mixer 24, and while both liquids were flowing in, they were mixed by inversion.

The mixing time was 6 minutes, during which time both solutions were mixed for 50 d.
There was an influx of people. At this time, the cock 25 was kept closed. As a control, an experiment was also conducted in which the reaction tank 23 was allowed to stand still (both liquids were flowed in from the bottom of the bag) (hereinafter, the mixed solution will be referred to as a treatment example, and the one left still will be referred to as a control).
After the reaction mixing operation was completed, the outflow operation was immediately started.

Open the cock 25 and use the pump 64 to pump 15 ml of the reacted liquid with the outlet facing down for both the treatment example and the control! /w
It was leaked in. Fractions of each of the five components were collected using a fraction collector 90, and the absorbance at 280 nm was examined. The results are shown in Table 1. In the control, unmixed B5A1 solution was detected at large fraction numbers, but in the treated example, no unmixed BSA solution was detected even at large fraction numbers.

Table 1 (blank below) Transplantable cancer VX2 was subcutaneously transplanted into the backs of nine domestic rabbits. 1
On the 5th day, a tumor of approximately 2c+aX2c■ was found by palpation. On the 16th day, an administration experiment was conducted on three domestic rabbits using the circuit shown in FIG. Each bomb 61.62, 22.6
For No. 3, a Pharmacia Verister pump was used. For tubes 71, 7273, 74, and 75, sterilized vinyl chloride tubes were used. The plasma separation means 1 includes a plasma separation means CR.

flo Type H-2 (Asahi Medical Co., Ltd.), the dialyzer 300 is the Microflo Type H-2 mentioned above.
The hollow fiber portion of Asahi Medical Plasma Flow AP-0311 (Asahi Medical Co., Ltd.) was filled with the housing. Steam sterilized 3M Na is used for plasma processing solution.
A Cl solution was used. The plasma processing reaction tank 23 contains blood.
Ng (Terumo Separation Ba Ng Teru Frenks 150mf.

Chill Shichisha) was used. Plasma separator l before extracorporeal perfusion
The internal portion of the hollow fiber, the dialyzer 300, and each portion of the tubes 71, 72, 73, and 75 of the reflux means were primed with physiological saline. 511+4 from rabbit auricular artery 51! /wi
n led the blood into the circuit. 0.3ml of heparin (blood clotting inhibitor) from the heparin supply device 52! /win was added. Plasma pump 62, plasma processing liquid pump 22
is operated on llR1/win and the reaction time is 20m1n
And so. Cock 25 must be closed in advance.2011
Released after in. Physiological saline was used as the external dialysis fluid and was refluxed into the dialyzer 300 at 100 me/s+in. After dialysis, the plasma was mixed with blood cells in the mixer 4, protamine was added from the protamine supply device 53 to neutralize heparin, and the blood was returned to the auricular vein 54 on the opposite side. A rocker platform for DNA extraction (Shiraki Seisakusho) was operated as a mixer 24 during the reaction period. After the administration experiment was completed, the rabbits were continued to be observed and the average survival days after tumor implantation were determined, and the results shown in Table 2 were obtained.

Of the nine rabbits transplanted with VX2 in Example 1, three of the rabbits that were not used in the administration experiment in Example 1 were tested without conducting an administration experiment using a circuit. Observation was continued, and the average number of days of survival after tumor transplantation was determined. This result is also shown in Table 2.
It was shown to.

Comparison■↓21 Among the rabbits transplanted with VX2 in Example 1, the mixer 24 was not operated for the three rabbits that were not used in Example 1 and Comparative Example 1-1 (i.e., the plasma processing During the reaction,
The experiment was conducted in the same manner as in Example 1, except that no mixing operation was performed, and the average survival days after transplantation of the Mffl tumor was determined. The results are also shown in Table 2.

Table 2 From Table 2, it can be seen that the average number of days of survival was longer for those in which the mixing operation was performed during the plasma treatment reaction.

Cunning■1 Using the same BSA solution as used in Experimental Example 1,
A mixing experiment was conducted using the apparatus shown in FIG.

The above-mentioned BSA solution was placed in the container 50, a calibrated chamber (Terumo quantitative infusion set, Chill Shichisha) was used as the plasma processing reaction tank 23, and a 3M sodium chloride aqueous solution was placed in the plasma processing liquid tank 21. I put it in. An aqueous sodium chloride solution and a BSA solution were supplied to the reaction tank 23 at a flow rate of 8 d/sin using a pair of Verister pumps 22.

As a control, an experiment was carried out in which the reaction tank 23 was left still (both liquids were flowed in from above the chamber), and as a processing example, a rocker platform for DNA extraction (Shiraki Seisakusho) was used as the mixer 24, and both liquids were flowed in. An experiment was conducted in which the mixture was mixed for a while. In both cases, the pressure inside the chamber was made equal to atmospheric pressure by opening the cock 26. The mixing time was 6 minutes, during which time both solutions were introduced at a rate of 50 d.

At this time, the cock 25 was closed. Immediately after the zero reaction mixing operation was completed, the same operation as in Experimental Example 1 was performed. The results are shown in Table 3.

As a result, as in Experimental Example 1, it was found that the treatment example in which the mixing operation was performed was better mixed.

Table 3 A graduated chamber used in Experimental Example 2 instead of the blood bag (Terumo Separation Bag Chillflex 150d, Chill Shichisha) in the plasma processing reaction tank 23 in the circuit shown in Figure 1 used in Example 1. (The reaction tank 23 and the cock 26 shown in FIG. 3, and the conch 26 were released through a sterile filter to make the pressure inside the chamber the same as atmospheric pressure.) A circuit was created.

An experiment was conducted in the same manner as in Example 1, except that the above circuit was used in place of the circuit in Example 1, and the average survival days were determined. The results are shown in Table 4.

In comparison, among the nine rabbits transplanted with VX2 in soil Example 2, the other three that were not used in the administration experiment in Example 2 were observed without conducting an administration experiment using a circuit. Next, the average number of days of survival after tumor transplantation was determined. This result is also shown in Table 4.
It was shown to.

Comparison (1) Of the rabbits transplanted with VX2 of Example 2, the mixer 24 was not operated for the three rabbits that were not used in Example 2 and Comparative Example 2-1 (i.e., the plasma processing reaction During,
The experiment was conducted in the same manner as in Example 2, except that the mixing operation was not performed, and the average number of days of survival after transplantation of 11!1ffi was determined. The results are also shown in Table 4.

Table 4 From Table 4, it can be seen that the average number of days of survival was longer for those in which the mixing operation was performed during the plasma treatment reaction.

Back layer (1) The experiment was carried out in the same manner as in Example 1, except that 3M sodium citrate aqueous solution was used as the plasma treatment solution instead of 3M NaCl aqueous solution, and the average survival days were determined. The results are shown in Table 5.

Northern comparison■Uni↓ Among the nine rabbits transplanted with VX2 in Example 3, three of them that were not used in the administration experiment in Example 3 were observed without conducting an administration experiment using a circuit. The average number of days of survival after tumor transplantation was determined. This result is also shown in Table 5.
It was shown to.

Among the rabbits transplanted with VX2 of Example 3, the mixer 24 was not operated for the three rabbits that were not used in Example 3 and Comparative Example 3-1 (i.e., the plasma processing reaction During,
The experiment was conducted in the same manner as in Example 3, except that the mixing operation was not performed, and Ilf! The average number of days of survival after tumor transplantation was determined. The results are also shown in Table 5.

Table 5 From Table 5, it can be seen that the number of days of survival was longer for those that underwent a mixing operation during the plasma treatment reaction.

In the previous Example 2, 0.1 M containing 0.15 M NaCl was used instead of the 3 M NaCl aqueous solution as the plasma treatment solution.
The experiment was conducted in the same manner as in Example 2, except that an acetic acid aqueous solution was used, and the average survival days were determined.

The results are shown in Table 6.

Comparison 1 (Nu) Of the nine rabbits transplanted with VX2 in Example 4, three were not used in the administration experiment in Example 4, and were observed without conducting an administration experiment using a circuit. The average number of days of survival after tumor transplantation was determined.The results are also shown in Table 6.
It was shown to.

Comparison Basis (21) Among the rabbits transplanted with VX2 in Example 4, the mixer 24 was not operated for the three rabbits that were not used in Example 4 and Comparative Example 4-1 (i.e., the plasma processing During the reaction,
The experiment was conducted in the same manner as in Example 4, except that the mixing operation was not performed, and the average number of days of survival after tumor transplantation was determined.

The results are also shown in Table 6.

Table 6 From Table 6, it can be seen that the number of days of survival was longer for those that underwent a mixing operation during the plasma treatment reaction.

(Effects of the Invention) By using the mixer in this manner, plasma and plasma processing liquid are mixed uniformly and quickly, so that malignant tumors and the like can be efficiently treated.

The basic operation method is to process the patient's blood and return it, so it does not put a burden on the patient's body like a surgical operation, and there is almost no loss of plasma proteins during the process. Since the circuit is isolated from the outside environment, it is safe and free from contamination by germs.

Using this circuit, patients who are difficult to undergo surgery or patients with malignant tumors for whom anticancer drug administration is inappropriate can be continuously and effectively treated.

FIG. 1 is an explanatory diagram showing one example of a circuit of the present invention, and FIGS. 2 and 3 are explanatory diagrams showing an experimental example apparatus of the present invention.

■ -・- Plasma separation means, 4- Mixer, 5- Rabbit, 21
-Plasma processing liquid tank, 22-pump, 23--plasma processing reaction tank, 24...mixer, 61, 62.63 pump, ?
1,72,73,74.75-tube, 200...
Plasma processing means, 300.--Adjustment means, 600.--Reflux means.

No. figure

Claims (1)

[Scope of Claims] 1. A plasma separation means for separating plasma from blood collected from a living body, and separating the separated plasma from a group of plasma processing solutions consisting of an inorganic salt solution, an organic salt solution, and an acidic solution. Plasma processing means for processing by contacting with at least one selected species; conditioning means for adjusting the treated plasma to the environment of the living body; and continuous or intermittent introduction of the adjusted plasma into the living body. and a reflux means capable of refluxing the blood plasma, and the plasma processing means is equipped with a mixer for promoting contact between the plasma and the plasma processing solution. circulation circuit.