JPH10276790A - Synthetic method and purification method of anti-tumor substance by degradation of tumor cells and tumor products by enterobacteria and Ray bacteria - Google Patents

Abstract

(57) [Summary] (Corrected) [PROBLEMS] To provide a method for synthesizing and purifying an antitumor substance by decomposing tumor cells and tumor products by enterobacteria and Ray bacteria. A tumor and a tumor product of interest are Ray bacilli,
It is decomposed into Escherichia coli or the like to synthesize an antitumor substance (denaturation and growth inhibitory substance) for the target tumor. The above product is extracted and purified as follows. The culture solution is sterilized by heating, alcohol is added, filtration is performed, and the alcohol is evaporated and then filtered again to remove water-insoluble substances. Acetone was added and acetone coagulated matter was removed. Chloroform was added twice as much as the stock solution to remove chloroform coagulation, and then chloroform-soluble cold aggregates were extracted. To the above is added twice the amount of ether to transfer the ether-soluble matter. The stock solution thus obtained is butanol: glacial acetic acid: water =
Developed by paper chromatography descending method in 4: 1: 2 system and R
Obtain a ninhydrin reaction positive substance around f = 0.16

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is to synthesize antitumor substances (degeneration and growth inhibitory substances) against symmetric tumors by decomposing tumors and tumor products to be symmetrical into Ley bacteria, Escherichia coli and the like. It has not yet been reported that tumors and tumor products were decomposed into bacteria to synthesize antitumor substances. Conventional anticancer drugs are chemically synthesized or extracts from non-tumorous organisms. We believe that this approach can be used to help cancer patients.

2. Synthesis method. The tumor cells (Ehrlich tumor cells are used in this experiment) and 500 ml of a culture solution thereof are heat-sterilized, transplanted with the above bacteria, and cultured at about 36.5 ° C. for about one week.

Reason for temperature condition. When the bacteria were cultured at 30 ° C and 37
The function of the bacterium is different when cultured at ℃.

Example. Ray bacteria produce prodegiodin at 30 ° C. (red), but form white colonies at 37 ° C. (not produce prodegiodin), and the surface temperature of the living body is around 36.5 ° C.

Method for extracting the product The extraction and purification methods are listed below in order.

The culture solution is heat-sterilized, and alcohol is added (final concentration: 80%), followed by filtration through Toyo filter paper. The alcohol is evaporated and filtered again at room temperature to remove water-insoluble materials.

After the addition of acetone (final concentration of acetone: 80%), the solidified acetone is removed, and the acetone is evaporated off.

Chloroform was added twice as much as the stock solution, and the mixture was shaken well to remove the chloroform coagulation. The aqueous layer was removed. Extract the chloroform soluble cold aggregates in water.

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 undiluted solution thus obtained is developed by paper chromatography in a butanol: glacial acetic acid: water = 4: 1: 2 system to obtain a ninhydrin reaction positive substance at around Rf = 0.16. 2m of the above substance-containing liquid
The experiment is carried out by creating a l). A crystal structure like a photograph attached is shown. The properties of the substances discovered this time are summarized as non-coagulable, soluble in alcohol, acetone and chloroform, insoluble in ether, and cold aggregating during chloroform dissolution.

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] (Examples) A mouse on the third day of Ehrlich cell intraperitoneal transplantation is administered with 0.5 ml of the above extract by subcutaneous injection once every three days. 11.14 after Ehrlich cell transplantation.
17． On the day, ascites was collected and cells were observed by Giemsa staining. The above experimental procedure is illustrated below.

(Results) Cell destruction is observed as shown in the attached photo,
The nucleus where the cells are swollen and ruptured loosely binds. Conventional anti-tumor substances mainly suppress the growth of tumor cells,
The substance that has been filed this time is characterized by its main function of destroying tumor 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 biosynthesis of an antitumor substance by decomposing a tumor cell and its product into bacteria 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 here.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification code FI C12R 1:43)

Claims (3)

[Claims] 1. A method for decomposing tumor cells and tumor cell products by enterobacteria including Ray and Escherichia coli. 2. A method for purifying the antitumor substance produced in [1] and the antitumor substance 3. The chemical, physical properties and crystal structure of the antitumor substance produced in [1]