JP2629856B2 - Antitumor agent - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to an antitumor agent containing α-carotene as an active ingredient.

2. Description of the Related Art Problems to be solved by the prior art and the present invention Currently, surgical treatment, radiation therapy, chemotherapy (drug administration), and the like are generally performed as cancer treatment methods.

Among these, as chemotherapy, treatments that administer drugs that kill tumor cells by directly acting on tumor cells have been widely applied, and proposals for antitumor drugs used in this type of therapy have been made. Many.

However, drugs that act on this type of tumor cells are:
Since it acts on normal cells as well as kills tumor cells, it has a high therapeutic effect on cancer, but has the disadvantage of very strong side effects.

Therefore, as a new antitumor agent, a drug that enhances the immune ability of normal cells and indirectly suppresses the growth of tumor cells has attracted attention, and for example, krestin, interferon, and the like have been developed. Although these drugs have a weak effect on tumor cells, they have the advantage of having few side effects.

More recently, decancerization of tumor cells (de-carcinogenesi
s), that is, normal cancer cells are not irreversible but reversible, and from the viewpoint that tumor cells lose their cancer characteristics and approach or return to normal cells, There is interest in applying drugs having a differentiation-inducing action to restore normal cells to cancer treatment. Although such drugs act directly on tumor cells, they have few side effects, are expected to have high therapeutic effects, and can be promising antitumor agents.

Currently, c-AMP is a drug having a differentiation inducing action.
Although prostaglandin E ₁ or the like having a function of increasing the amount has been proposed, for prostaglandin E ₁ is unstable in the human body, when using the cancer treatment must drip into the affected area directly In addition, there is a problem that it needs to be used in combination with other drugs. Therefore, a substance which has an excellent differentiation-inducing effect, is stable, has low toxicity, and is effectively used as an antitumor agent is desired.

Means and Actions for Solving the Problems The present invention has been studied diligently for substances having a differentiation-inducing action and having an excellent antitumor activity. As a result, α-carotene has a strong tumor cell growth inhibitory effect, Has been found to be excellent in inducing differentiation into normal cells, and can be effectively used as a so-called decancer, and have accomplished the present invention.

Heretofore, it has been known that β-carotene has an antitumor activity in a lymphocyte system (“Oncology 39: 33-37”).
(1982) "and" BIOCHEMICL AND BIOPHYSICAL RESEARC "
H COMMUNICATION, 1130-1135 (1986) ”)
There is no report that carotene has antitumor activity. As is clear from the description of the experimental examples described later, β-carotene has almost no tumor cell growth inhibitory effect,
-Carotene has a remarkable effect, has excellent properties as an antitumor agent, and also has an excellent decancer effect as described above, and is effectively used in chemotherapy for returning tumor cells to normal cells. This relates to a new finding of the inventor.

 Hereinafter, the present invention will be described in more detail.

The antitumor agent of the present invention contains α-carotene as an active ingredient.

Here, α-carotene may be a synthetic product or a natural product. Natural α-carotene is usually present as a mixture with various carotenes such as β-carotene and r-carotene,
In the present invention, α-carotene can be blended in a state of a mixture with such various carotene. As natural carotene, carotene extracted, concentrated and purified from palm oil is particularly suitable. The carotene obtained from this palm oil is a mixture of carotene such as α, β, etc., and about 25 to 40% of α-carotene, about 50 to 70% of β-carotene, and about 10% or less of other components such as carotene oxide. Is a red opaque paste. This natural carotene has high solubility in oil and has good absorption in the body when administered to the human body.

Furthermore, in the present invention, a mixture of various carotene can be purified and isolated α-carotene can be used. In particular, when isolated α-carotene is used as the active ingredient of the present invention, a stronger tumor cell growth inhibitory effect and differentiation inducing effect are obtained than when a mixture of carotene containing the same amount of α-carotene is used. be able to.

In addition, the method of purifying and isolating α-carotene may be an ordinary method.

The antitumor agent according to the present invention contains α-carotene as an active ingredient as described above, and a carotene mixture containing α-carotene or α-carotene may be used alone or in combination with other pharmaceutical ingredients as necessary. It is administered by intravenous injection, subcutaneous injection, intramuscular injection, oral administration, rectal administration using suppositories, and the like. The dose varies depending on the route of administration, the number of administrations, etc.
In addition, it can be changed widely depending on the severity of the symptoms and the like, for example, 15 μg to 3 g / kg body weight or 1 mg to 1 day per adult.
Can be 10g.

When the antitumor agent according to the present invention is formulated, α-
An effective amount of carotene may be mixed with an appropriate amount of a non-toxic carrier, and prepared for administration using any conventional formulation method. That is, when prepared for oral administration, soft capsules,
Hard capsules, tablets, granules, fine granules, powders, sustained release agents for active ingredients, solutions, suspensions, etc. For parenteral administration, injections, drops, narcotics, etc. . In this case, in formulating the preparation, a non-toxic carrier such as a surfactant such as sucrose fatty acid ester, fatty acid monoglyceride, propylene glycol fatty acid ester, sorbitan fatty acid ester, or lecithin; Binder, sucrose, lactose, starch, crystalline cellulose, mannitol, light anhydrous silicic acid, magnesium aluminate, magnesium metasilicate aluminate, synthetic aluminum silicate, calcium carbonate, sodium hydrogen carbonate,
Excipients such as calcium hydrogen phosphate and calcium carboxymethylcellulose, lubricants such as magnesium stearate, talc, hydrogenated oil, salt, saccharin, orange oil, licorice extract, citric acid, glucose, menthol, eucalyptus oil, malic acid Flavoring agents, flavoring agents, coconut oil, olive oil, sesame oil, peanut oil, soybean oil, medium chain fatty acid triglycerides, safflower oil, suspending agents such as soybean phospholipids, wetting agents, cellulose acetate phthalate (CAP), etc. Carbohydrate derivatives such as cellulose and sugars, acrylic copolymers such as methyl acrylate / methacrylic acid copolymers, methyl methacrylate / methacrylic acid copolymers, and polyvinyl derivatives such as diester monoesters and the like It is prepared by a conventional method using components such as a film forming agent and a coating aid. It is subjected to. Preparations for mucosal application and also injections are prepared by conventional methods. However, when a method of suspending or emulsifying in distilled water for injection is adopted, soybean oil, peanut oil, medium Chain fatty acid triglycerides and the like can be used, and sucrose fatty acid esters, fatty acid monoglycerides, propylene glycol fatty acid esters, sorbitan fatty acid esters, lecithin and the like can be used as emulsifiers.

Effect of the Invention α-carotene used as an active ingredient in the antitumor agent according to the present invention has a strong tumor cell growth inhibitory effect and has an excellent differentiation inducing effect of returning tumor cells to normal cells. It is preferably used. Furthermore, this antitumor agent containing α-carotene as an active ingredient has low toxicity, and is 10 g / kg body weight (3.5 g in terms of α-carotene) when administered as palm carotene in mice.
/ kg body weight). Furthermore, since it is relatively stable in the body, it is also possible to administer a large amount of it alone to the human body without using it in combination with other drugs. It is used effectively for the treatment of primary tumors such as cervical cancer, and is particularly effective for neuroblastoma.

Hereinafter, the effects of the active ingredient of the present invention will be specifically shown by experimental examples.

[Experimental Example 1] Tumor cell growth inhibitory effect Human neuroblastoma (GOTO cells) was seeded on a dish having a diameter of 35 mm at a ratio of 4 × 10 ⁴ cells / 2 ml medium / dish and cultured for 2 days.

Separately, 2.15 volumes of a 1% emulsion of palm carotene extracted, concentrated and purified from palm oil was mixed with 7.85 volumes of 95% ethanol and sterilized.

Next, palm carotene 10μ thus sterilized
(Α-carotene 6μ out of final concentration 20μM palm carotene)
M, β-carotene (14 μM) was added to the above dish that had been cultured for 2 days, and the culture was continued for another 5 days. As a control, the same operation was carried out for 10 μl of the same emulsion / 95% ethanol (volume ratio: 2.15 / 7.85) except that carotene was not contained.

After the cultivation, the number of tumor cells in each dish was measured, and the ratio of the number of tumor cells to the control group was determined to examine the effect of palm carotene on tumor cell growth inhibition (measurement number twice).

In addition, instead of palm carotene, α-carotene (2 μM) and β-carotene (2 μM) treated in the same manner as palm carotene were respectively added to the above dishes and cultured in the same manner, and the effect of each carotene on tumor cell growth inhibition was examined. Was.

Differentiation Inducing Effect The morphological changes of GOTO cells in the dish to which the above α-carotene (2 μM) was added and the control dish were observed, and the induction rate of α-carotene to induce tumor cell differentiation was determined. In addition, the differentiation induction rate is the one that can measure the length of the protrusion in the cells in the dish.
500 cells were observed, and those having a protrusion length of twice or more the major axis of the cell body were determined as differentiated cells, and determined according to the following formula.

The above results are shown in Tables 1 and 2.

From the results in Table 1, it was confirmed that palm carotene containing α-carotene and β-carotene exhibited an inhibitory effect on the growth of tumor cells (GOTO cells).

Also, from the results in Table 2, β-carotene showed only a slight tumor cell growth inhibitory effect, whereas α-carotene showed a stronger tumor cell growth inhibitory effect as compared to β-carotene. It was confirmed that α-carotene was mainly involved in the same effect of palm carotene.

Furthermore, from the results in Table 2, it was confirmed that in the cultured cells to which α-carotene was added, neuroblast-specific projections appeared, and the tumor cells were differentiated into normal cells.
M) revealed that morphological differentiation was induced.

The morphology of GOTO cells (neuroblasts) to which α-carotene has been added is as shown in Reference Figures 1 and 2. here,
Reference FIG. 1 shows the form of the control group, and FIG. 2 shows α-carotene 2 μm.
5 is a micrograph (400-fold magnification) showing the morphology of a group to which M was added and cultured for 5 days. From the reference figure, it can be seen that α-carotene returns tumor cells to normal cells.

[Experimental Example 2] Using the human tumor cells shown in Table 3 and using α-carotene and β-carotene at a ratio of 20 μM, the same operation as in Experimental Example 1 was performed to measure the number of tumor cells. The tumor cell growth inhibitory effects of α-carotene and β-carotene were evaluated by determining the ratio of the number of tumor cells to those without any of α-carotene and β-carotene (blank). The results are shown in Table 3.

From the results in Table 3, it can be seen that α-carotene effectively suppresses the growth of various tumor cells and acts effectively on various tumor cells as compared with β-carotene.

[Experimental Example 3] Using human tumor cells shown in Table 4, α-carotene was
Using the ratio of μM, the number of tumor cells was measured in the same manner as in Experimental Example 1, and the ratio of the number of tumor cells to the control group (one to which neither α-carotene nor β-carotene was added, blank) was determined. The effect of α-carotene and β-carotene on tumor cell growth was evaluated. The results are shown in Table 4.

From the results in Table 4, it can be seen that α-carotene effectively inhibits the growth of various tumor cells regardless of their germ source, that is, whether they are ectodermal or endodermal, and that solid tumors are also leukemia Is also found to be effectively treated with α-carotene.

[Experimental Example 4] 30 mice were divided into 15 mice per group, and an acetone solution of dimethylbenzoanthracene (DMBA) was applied to the depilated back skin of each mouse (100 µg DMBA / mouse). One week later,
As a control, one group of mice received TPA (12-0-T
Acetone solution of etradecanogl Phorbol 13-acetate) was applied twice a week (0.5 μg TPA / mouse / time), and the other group was given a TPA acetone solution supplemented with palm carotene twice a week.
Apply twice (162 nmol palm carotene / mouse / times, TPA
The amount of application was the same as described above), and the carcinogenesis rate was examined over time. The results are shown in the drawing.

Note that the same palm carotene as in Experimental Example 1 was used.

From the results shown in the drawings, it is confirmed that palm carotene has a favorable skin carcinogenesis inhibitory effect. In addition, all the mice were alive during the above experimental period.

Next, Formulation Examples of the antitumor agent of the present invention will be described with reference to Examples.

Example 1 Capsule 100 g of palm carotene was suspended in 1 kg of wheat germ oil,
Fill into individual gelatin capsules. One per day
Administer ~ 10 orally.

Example 2 Injection Palm Carotene 100 g Stearic acid monoglyceride 100 g Peanut oil 200 g Sucrose glycerin stearin ester 50 g Ascorbic acid stearate 20 g Distilled water for injection 9530 g Prepare an injection with the above composition and fill each ampoule 10 ml.

Example 3 External ointment Polyethylene glycol 4000 150 g Polyethylene glycol 400 150 g α-carotene 1 g The above composition was humidified and mixed, and used as an ointment for skin cancer.

[Example 4] Suppository Methyl salicylate 0.0350 g Famazole T-115 (Nissan Chemical Co., Ltd.) 2.0000g α-carotene 0.0100g

[Brief description of the drawings]

FIG. 1 is a graph showing the effect of palm carotene on skin cancer suppression.

Claims (1)

(57) [Claims] 1. An antitumor agent comprising α-carotene as an active ingredient.