JP3088680B2 - Synthetic method of antitumor substance by intestinal bacteria and ray bacteria - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention
A method for synthesizing an antitumor substance.

[0002]

BACKGROUND OF THE INVENTION Conventional anti-cancer drugs are chemically synthesized or are extracts from non-tumor organisms. It has not yet been reported that tumors and tumor products were decomposed by bacteria to synthesize antitumor substances.

[0003]

DISCLOSURE OF THE INVENTION According to the present invention, tumors and tumor products to be symmetric are decomposed by intestinal bacteria such as Ley bacteria and Escherichia coli to synthesize antitumor substances (denaturation and growth inhibitory substances) against symmetric tumors. That is. This technique can be used to help cancer patients.

[0004]

The antitumor according to the first aspect of the present invention.
The method of synthesizing the substance is based on Ehrlich tumor cells and their culture solution.
Is heat-sterilized and transplanted with intestinal bacteria and Ley bacteria at around 36.5 ° C.
For about one week.

[0005]

DESCRIPTION OF THE PREFERRED EMBODIMENTS [Synthesis method. 36. Tumor cells (Ehrlich tumor cells are used in this experiment) and 500 ml of a culture thereof are heat-sterilized, and the above bacteria are transplanted.
Incubate at about 5 ° C for about one week. [Reason for temperature condition. ] When the bacteria were cultured at 30 ° C and 3
When cultured at 7 ° C., the function of the bacteria is different. Example. Ray produces prodigiosin at 30 ° C (red)
At 37 ° C., produce white bacterial colonies (no prodigiodin). The surface temperature of the living body is about 36.5 ° C.

[Production Extraction Method] Hereinafter, extraction and purification methods will be exemplified in order. The culture solution is heat-sterilized, alcohol is added (final concentration: 80%), and the solution is filtered through Toyo filter paper. The alcohol is evaporated and filtered again at room temperature to remove water-insoluble materials. After addition of acetone (acetone final concentration: 80%), acetone is removed by coagulation, and the acetone is evaporated off. After adding chloroform twice as much as the stock solution and shaking well to remove chloroform coagulation, the aqueous layer was removed, and the chloroform layer was placed at a temperature of 10 ° C. or lower, and the formed cloudy coagulation was separated by filter paper. The filter paper is placed in water to extract chloroform-soluble cold aggregates. Ether is added twice as much as the chloroform-containing solution containing the cold aggregate, and the mixture is shaken well to transfer the ether-soluble matter, and then the ether is removed. The stock solution thus obtained is developed with a butanol: glacial acetic acid: water = 4: 1: 2 system by a paper chromatography descending method to obtain a ninhydrin reaction-positive substance around Rf = 0.16 (raw material stock solution 500).
When the amount is ml, prepare 2 ml of the above-mentioned substance-containing solution and conduct the experiment). In addition, the crystal structure as shown in the attached photographs (see FIGS. 1 and 2) is shown.

[0007] The properties of the substance discovered this time can be summarized as follows: non-coagulable, soluble in alcohol, acetone and chloroform, insoluble in ether, and cold-coagulating in chloroform. The most characteristic feature of the above extraction method is that a method of cold-aggregating chloroform-soluble matter has been found.

[Examples of Use and Results] (Example) A mouse (experimental body) on the third day of Ehrlich cell intraperitoneal transplantation is injected with 0.5 ml of the above extract by subcutaneous injection once every three days. After Ehrlich cell transplantation 1
1.14. Ascites was collected on day 17 and the cells were observed by Giemsa staining. The above experimental procedure is shown in FIG .

(Results) Photographs attached to cell destruction (FIG. 4)
See FIG. ), And the nucleus in which the cells are swollen and ruptured loosely binds.

The photograph in FIG. 4 shows that the cells are stained uniformly.
Is shown (symmetric). The photograph in FIG.
On the second day after three administrations of the strike solution, the cell membrane swells and
Is shown. The photograph in FIG. 6 shows the test solution
5 days after administration, swelling of cells became remarkable, and
The cells are shown torn. Picture of FIG.
Is on the 8th day after three administrations of the test solution.
Ruptures without shrinkage, with debris visible at center and right end
It is. The photograph in FIG. 8 shows that the cell membrane has been lost and the nucleus has been decomposed.
The state is shown.

[0011] Conventional antitumor substances mainly suppress the growth of tumor cells, but the substance according to the present application is characterized in that it mainly acts to destroy lung ulcer cells. Note that the amount of tumor cells generated is suppressed to １ ／ or less of the symmetric group. No degeneration of various other organs of the experimental body was observed.

As described above, there is no report on a method of biosynthesizing an antitumor substance by decomposing a tumor cell and its product by a bacterium with respect to a target tumor cell.
Although a method for purifying materials using chemical properties has already been established, the target substance was obtained using the property of being soluble in chloroform and causing cold aggregation, as described in the extract method. This is a major feature of the purification method.
We believe that a new field of antitumor therapy will be opened using the synthetic and purification methods claimed herein.

[Brief description of the drawings]

FIG. 1 is a diagram showing a micrograph showing a columnar crystal.

FIG. 2 is a diagram showing a micrograph showing a state in which crystals are accumulated.

FIG. 3 is a diagram showing a procedure of a usage example.

FIG. 4 shows a symmetric micrograph showing the stained cells.

FIG. 5 is a diagram showing a micrograph of cells two days after administration of a test solution.

FIG. 6 is a diagram showing a micrograph 5 days after administration of a test solution.

FIG. 7 is a diagram showing a micrograph on the 8th day after test solution administration.

FIG. 8 is a micrograph of a cell whose cell membrane has been lost and whose nucleus has been decomposed.

──────────────────────────────────────────────────の Continued on front page (51) Int.Cl. ⁷ Identification code FI (C12P 1/04 C12R 1:43)

Claims (1)

(57) [Claims] 1. An Ehrlich tumor cell and its culture solution
Sterilize by heating, inoculate with intestinal bacteria and ray bacteria
A method for synthesizing an antitumor substance that is cultured for about one week.