JPH02240019A - Anti-carcinogenic promoter - Google Patents

Abstract

PURPOSE:To obtain an anti-carcinogenic promoter useful as a drug to prevent carcinogenesis, healthy food or functional food, comprising alpha-monolinolein as an active ingredient. CONSTITUTION:An anti-carcinogenic promoter comprising alpha-monolinolein having extremely inhibitory action on manifestation of early antigen of Epstein Barr virus by tetradecanoylphorbol acetate (carcinogenic promoter) as an active ingredient. alpha-Monolinolein is obtained by separation and purification from a vegetable oil such as soybean oil, adlay oil or seed cotton oil, esterification or ester exchange of glycerin using acid catalyst or enzyme or hydrolysis of triglyceride or diglyceride. alpha-Monolinolein is considered to be used as an antiviral agent and a carcinostatic agent since alpha-monolinolein inhibits manifestation of Epstein Barr virus.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to an anti-carcinogenic promoter used for the purpose of preventing carcinogenesis.

(Prior art and problems to be solved) The two-stage theory of carcinogenesis, which is generally accepted as a carcinogenic mechanism, is that an initiator damages the chromosomes of normal cells;
It is based on an experimental system that generates tumors through the action of promoters that exert an epigenetic effect on cells that have undergone this disorder and have turned into potential tumor cells, transforming them into tumor cells.

DMBA (dimethylbenzanthracene) and the like are known as typical initiators, and TPA (tetradecanoylphorbol acetate) and the like are known as promoters. Research is underway to clarify the mechanism of carcinogenesis through carcinogenesis experiments using these promoters and initiators.

In particular, the process of promotion caused by promoters is considered important in conjunction with the universality of oncogenes, and there are many attempts to derive new concepts of anticancer properties by clarifying this point. One of these is the search for anti-carcinogenic promoters that inhibit the action of oncogenic promoters, and so far we have used glue poxygenase inhibitors such as NDGA (nordihydroguaiaretic acid), calmodulin inhibitors such as W-7, glucocorticoids, and retinoids. However, the search for such inhibitors is still in an underdeveloped area.

It is also a naturally occurring anti-carcinogenic promoter. Many attempts have been made to explore the potential of carcinogenesis, and crude plant extracts are known to inhibit experimental carcinogenesis, but there are few examples of their active ingredients being clarified.

The problem to be solved by the present invention is to discover a compound (anti-carcinogenic promoter) from the plant kingdom that has a carcinogenic promotion inhibiting effect, and to provide a carcinogenic promotion inhibitor containing this compound as an active ingredient.

(Means for Solving the Problem) The present inventors inhibited the expression of the EBV (Epstein-Barr virus) genome in the Raji strain, which is a cultured cell derived from Purkitt's lymphoma and contains the EBV (Epstein-Barr virus) genome. Focusing on the fact that many of the compounds shown to act as anti-carcinogenic promoters in a two-stage mouse skin carcinogenesis experiment, we searched for virus/genome inactivating substances that inhibit EBV/genome expression among components contained in plants. The inhibitory effect on the expression of the EB virus genome is observed by the following method.

That is, in the Raji strain culture system mentioned above, n, which acts synergistically with TPA, a carcinogenic promoter, to express activity.
Monobutyric acid and the test substance were added and cultured, and EBV-EA (EBV-EA), which is activated by TPA and expressed on the cell surface, was cultured.
This is a method for detecting B. virus (early antigen) using an indirect fluorescent antibody method using antibodies derived from serum from patients with nasopharyngeal cancer.

Specifically, EBV latently infected human lymphoblastoid cell line R
RPMI 1640 with fetal serum and antibiotics added is used as a culture medium for aji cells.

Under this culture condition, EBV-E. 'l. Natural occurrence rate is 0
．． It is less than 1%. Raji cells adjusted to a concentration of I x 106 cells/ml were treated with 4mM n-monobutyric acid, 20ng
/ml of TPA and 50 times the molar amount of α-monolinolein to TPA in the above culture solution at 37°C.
, incubate for 48 hours. EBV-EA was stained using indirect fluorescent antibody method using nasopharyngeal cancer patient serum, and the percentage of positive cells was determined by α.
- Calculated against the control without monolinolein added and used as the viral genome expression inhibition activity.

The virus genome expression inhibition activity of α-monolinolein is 100%.

As described above, since the α-monolinolein of the present invention significantly inhibits the expression of EBV-EA by TPA, it can be expected to act as an anti-carcinogenic promoter.

The inhibitory effect of α-monolinolein on carcinogenesis can be clearly demonstrated through a two-step mouse skin carcinogenesis experiment.
Specifically, the following method is used.

First, the carcinogenesis promoting effect was found in the dorsal skin of mice.
, 12-dimethylbenzanthracene (DMBA)
After one week, promotion activity is examined by applying TPA twice a week.

On the other hand, in order to investigate the inhibitory effect on carcinogenesis promotion, TP
One hour before applying A, the test compound is applied and compared with the promotion activity of the control with only TPA applied.

Promotional activity is evaluated by the proportion of mice bearing tumors and the average number of tumors per mouse.

For example, in the case of α-monolinolein, DMBA+TP
While group A reached a tumor incidence of 100% after 8 weeks, the tumor incidence in the DMBA+α-monolinolein+TPA group pretreated with α-monolinolein was 40% even at 11 weeks. In addition, when comparing the average number of tumors per mouse, the DMBA + TPA group was 4.8 at week 13.
, whereas DMBA 10-monolinolein + T
The PA group had 2.7 pieces.

The above results indicate that α-monolinolein is an anti-carcinogenic promoter that significantly inhibits carcinogenic promotion by TPA.

α-Monolinolein, which is an anti-carcinogenic promoter of the present invention, can be regulated by the following method.

For example, inumameike, pearl barley oil,
It can be separated and purified from vegetable oils such as cottonseed oil and rice nougat oil. Further, known synthetic methods such as esterification and transesterification of glycerin using an acid catalyst or an enzyme, and partial hydrolysis of corresponding triglycerides and diglycerides can also be used.

The anti-carcinogenic promoter of the present invention is α. It can be used in the form of monolinolein or a composition containing it as an active ingredient in an effective and non-toxic amount.For example, as an oral preparation, solid forms such as tablets, capsules, troches, granules, powders, etc. It can be used as a preparation or a solution such as a solution or syrup. These various preparations can be manufactured by known methods using conventional inorganic or organic, solid or liquid pharmaceutical preparations.

(Example) Below, the present invention will be described in more detail based on Examples.

Example 1 87 g of oil was obtained by immersing 2 kg of threshed pigeon berry seeds in hexane, filtering the resulting leachate, concentrating the hexane, and distilling off the hexane.

Next, this was partitioned between hexane and methanol to obtain 39 g of oil from the hexane layer, which was subjected to silica gel column chromatography using hexane/ethyl acetate as the eluent to separate triglyceride fraction, diglyceride fraction, free fatty acid fraction, It was fractionated into a sterol fraction and a monoglyceride fraction.

The monoglyceride fraction was further purified by silica gel column chromatography to separate α-monolinolein and α-monolinolein.
．． It was possible to separate it into monoolein.

Thus isolated α. Monolinolein showed the following physical constants in various spectra measurements, and these values agreed with those of standard α-monolinolein.

IR (V): 1744 cm-1 LH-NMR (δppm): 5.34 (4H),
4.17 (2H), 3.93 (IH), 3.70 (L
H), 3.59 (LH), 2.79 (2H), 2
．． 35 (4H), 2.04 (4H), 1.63 (2H
), 1.31-1.25 (IOH), 0.88 (3
H) in CDC1313C-NMR (δppm)
: 174.3, 130.1, 129.7, 12
8.1, 127.9, 70.3, 65.2, 63
．． 4, 34.2, 31.9, 29.8, 29.
7, 29.5, 29.3, 29.3, 29.1,
27.2, 24.9, 22.7, 14.1 i
n CDCI3FD-MS (m/z): 354 (M
), 355 (M + H) Example 2 Three groups of 7-week-old female ICR mice (5 mice per group) were used, and 390 nmol of DMBA per mouse was applied only once to the back skin. After one week, 1.7 nmol of TPA was applied twice a week to examine the promotional activity and this was used as a control. On the other hand, 501 lg of α-monolinolein was applied 1 hour before applying TPA.
Compare the promotional activity of the TPA-only control with the method described above.

Tumors occurred in the DMBA + TPA group from the 6th week and reached 100% tumor incidence in the 8th week;
-The frequency of tumor occurrence in the monolinolein-pretreated DMBA10-monolinolein+TPA group was 40% at the 11th week.

Furthermore, the average number of tumors per mouse was 4.8 in the DMBAfTPA group at week 13, whereas
A+a-mo/')/Re{>ten'r'pA/i
'P was 2.71 hard.

(Effects of the Invention) Therefore, the anti-carcinogenic promoter of the present invention containing α-monolinolein as an active ingredient can be used as a pharmaceutical, health food, or functional food for the purpose of preventing carcinogenesis.

Furthermore, since α-monolinolein, which is an active ingredient of the cancer promotion inhibitor of the present invention, inhibits the expression of the EB virus genome, it can also be used as an antiviral agent and an anticancer agent.

Claims (1)

[Claims] Anti-carcinogenic promoter containing α-monolinolein as an active ingredient