JPS6012071A - Body fluid treating agent having anti-tumor action - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] (Technical field of invention) TECHNICAL FIELD The present invention relates to a blood treatment agent having antitumor effects.

(Prior art and its problems) The number one cause of death in developed countries is due to cancer, but so far no cancer-fighting drugs as powerful as antibiotics have been found to fight infectious diseases. The ribopolysaccharides present in the cell walls of gram-negative bacteria are substances that are also called bacterial endotoxins, endotoxins, or pyrogens, and have a thermogenic effect.

-C is known to exhibit harmful effects such as Schwartzmann activity and lethal toxicity, but on the other hand, anti-II! It is also known as a substance that exhibits a tumor effect.

At present, there is no specific effect on malignant DI tumors, and substances with antitumor effects are worthy of attention, but in the case of the ribopolysaccharide,
Because its lethal dose is close to the minimum effective antitumor dose, it cannot be used safely.

(Objective of the invention) To attenuate the lethal toxicity of the ribopolysaccharide while maintaining its antitumor effect.

(Structure 2 of the Invention) The present invention provides a body fluid treatment agent in which a Gram-negative bacterial cell wall, the cell having endotoxin activity, and wall constituent components are immobilized within a semipermeable membrane or gel.

What is the cell wall component having Gram-negative bacterium 11111 cell Ap and En1-toxin activities in the present invention?

Gly\-negative cocci, represented by N eisseria gonorrboeae, P SeudOmOnaS
aeruginosa, 3 rucel 1aab
ortus, Bordetella pertussi
Gram-negative aerobic bacilli, such as F sc
bericl)iacoli, 3a1monella
tyl+l+i, 31+igelladyscnta
riae, K lel+5iclla 1+neu+
noniae.

3erratia marccscens, F'rot
cus vulgaris,, YOrSlnla el
lIel'0cOltUIca, V It) rloCl
Cell walls of Gram-negative bacteria such as Gram-negative facultative bacilli such as 1OIerae and e, and complexes of lipids and polysaccharides or complexes of lipids and polysaccharides and proteins localized in the outer layer of the cell walls. For example, Gram-negative bacteria can be cultured in a Hill homogenizer (with a Hill homogenizer). It can be made by centrifugation after it has been destroyed, and
Ribopolysaccharides (including complexes with proteins), which are typical examples of cell wall components with IS properties (1 and 1 hexin), can be extracted from Gram-negative bacteria by known methods. A typical example of this method is the method of extraction with 71Nol water (Van Otto Westphalat al.
,,Z,Naturforscl+,,7 B: 1
48-155 (1952)), method of extraction with lychloroacetic acid (A, Boivin and l-
, Mesero, 1) eanu, , Comp.

Rc++d,Soc, +3 iol,,12 B!
5 (19go(8)).

Gino method of extraction with butanol (1,), C, Mo
rriso++ and L, Leive, J.
B iol, cl+en32yi O(8)2911
(1975)), J3 and ethylenedino7mine-N
, N, N', N'-de1-method of extraction with acid-resistant water (L, Leive et al. 1.), Biol,
CI+em, .

2/+3 6384 (1968)).

In the present invention, the semipermeable membrane used for embedding cell wall components in Gram-negative bacterial cell walls is capable of eluating the cell walls and cell wall components.

In addition, it has the ability to pass plasma components, so it is suitable for use! If J3 is below 1 and is chemically, 13J, and physically stable, there are particular restrictions on 1j (yes.) Examples include (1) acrylamide 1~/meburene bisacrylamide copolymer. , (2) Poly”-dylenimine 1
Pichlorohydrin reactant, (3) isotuffic and syndi A° tactical polymethacrylate-1-.
Stereo complex, (4) Ionic complex of acrylic 12 ester copolymer containing sulfuric acid group and acrylic acid ester copolymer containing quaternary am7nium group, +5) Acid-resistant ester of cellulose, (6) Polymer Hinirun Full Call Conductor, OJ: Bi, (7) Acrylic 21
- There are semipermeable membranes or gels such as Lyle copolymers.

The preparation of the body fluid treatment agent of the present invention includes (1) Gram-negative bacterial cell wall or endo-1 to cell wall 411 having endoxin activity;
A method of forming a gel in a solution, turbidity, or emulsion of one component; or (2) dissolving a polymer in the liquid and extruding it into a non-solvent of the polymer; C)
The method for avoiding the formation of IIC is as follows: (a) the cell wall or the cell wall components are adsorbed onto the fibers or membranes already formed, or the cell wall components are coated with a gel d1. Alternatively, it can be constructed by (4) the 7J method in which the cell wall or the cell wall component is attached together with a gel onto the fiber or membrane. In (3) above.

Fibers made of a polymer having an isoelectric point of 7 or higher, preferably 9 or higher, or 11Q, are preferably used because they have a high affinity with the cell wall or the cell wall component.

If the density of the cell wall or the cell wall component in which the present invention 5 is immobilized in the body fluid treatment agent is too low, a large amount of body fluid treatment agent will be required, which is not preferable.

0.1n+g/g or more, more preferably lm+7g or more is required.

The shape of the body fluid treatment agent of the present invention is not particularly limited as long as it has a suitable surface area and sufficient shallow strength to withstand external forces during use.

Generally, 1-particle shape, I-fiber shape, hollow fiber shape, and membrane shape C are used.

Use of the body fluid treatment agent of the present invention: In the dino method, a column for extracorporeal circulation is filled with blood, and in the fJ method, blood is brought into contact with plasma continuously or intermittently. but.

After filling the inside of the syringe and aspirating the entire surface or plasma into it, refill the syringe with blood (31 plasma□
There are methods such as returning the body fluid treatment agent to the body, or leaving the body fluid treatment agent in the body.

(Mechanism of action of the invention) Although the antitumor effect of the body fluid treatment agent of the present invention is not a clear Mr. -U shimeru' () (1), and in this case, since the ribopolysaccharide is immobilized on the carrier, it is thought that there will be no lethality.

(Effects of the Invention) The body fluid treatment agent of the present invention is characterized in that it exhibits antitumor properties similar to lipopolysaccharide and does not exhibit a lifesaving effect.

Examples are shown below.

Example 1 T-ray sterilization (Co, 12.000 rad/min, 1 hour Terukawa) Escl+cricbia coli, 5t
rain B moisture 1711 pieces 3C1 120+nl F/
A. Float the glass beads (diameter 0.17 to 0.1
Shake for 3 minutes at below 5℃ with 120g of 8mm cell
00 cycles/min) destroyed. Beads were removed by glass filter filtration, and centrifuged at 1,200xg for 30 minutes.''The clear liquid was further centrifuged at IO,0OOXQ for 20 minutes to remove the supernatant.
Collect the cell walls and add O, iM phosphate M buffer (pl” 7.0).
After washing twice with IM-saline and once with distilled water,
Freeze-drying yielded cell wall 1Q.

50 ml of the above Ip11 cell wall was finely dispersed in 25 ml of 0.1M phosphate buffer (pH 7,4), and acrylamide IC1 was added to the same buffer ah 25 +111.
8 Hγ, J>, J: Bi, N, N-methylenebisino/kryl i) 2.5 g of the same buffer solution 10 ml
Add 0.111111 paste Bonolahin Q and 1 mg of potassium persulfate to the mixture and irradiate with high 1 mercury SO:I (500W) for 10 minutes under a nitrogen stream.
-IL/'cpolymerized I,:. The generated gel has a diameter of 0
．． 15) Crush and 60'C
Rinse 10 times with 500ml of warm water, and then add 0.1M
Phosphate buffer (++ H7,4>3(') 10 in Oml
After washing twice, the cell walls encapsulated in polyacrylamide foil were removed (11k).

Example 2 Fiber 811 sarcoma M induced by inoculating BAL 13/C mice with Medilni 1 lanthrene
J.
Tumor-bearing masks were prepared. The diameter of the canker grew to 2.1 mm 50 days after inoculation, 6.3 mm after 10 days, and 8.9 strokes after 12 days.

Add 2 liters of the cell wall-immobilized gel prepared in Example 1 to 60 ml of BΔ11-3/C mouse serum (dry weight, cells vB+
After shaking at 37°C for 180 minutes, centrifugation was performed to collect the C9 supernatant (activated serum).

Twenty-one tumor-bearing mice, 9 days after inoculation with one tumor cell, were prepared and divided into 3 groups of 7 mice each. Food j2. j
Diluted with water and administered intravenously once a day, 5 times over 5 days43
L, for the second group, 0.5 ml of the above activated serum was diluted with 2,5111 saline and 21t, I
, intraperitoneally administered 5 times at interval 1 (]A, and the third group was observed without any treatment. Results.

The average tumor diameter 45 days after inoculation with tumor cells was 4.7 mm and 24 mm in the first group, respectively.
．． 1mm, 2nd? 4.7mm and 21.8m for IY
m, 4 in the third group. ．． 8mm and 29.0mm.

In addition, tumor shrinkage was observed in 1 out of 7 animals in the 1st group, 3 out of 7 animals in the 2nd group, and O out of 7 G and t animals in the 3rd group. The gun that was refurbished was the 1st group J3.
There were 0 mice in the 3rd group and 2 mice in the 2nd group.

Claims (1)

[Claims] A body fluid treatment agent comprising a Gram-negative bacterial cell wall or a component of the cell wall that exhibits endotoxin activity immobilized within a semipermeable membrane or gel.