JPS63201130A - Antitumor agent - Google Patents

Abstract

PURPOSE:To provide an antitumor agent containing extract of Solanum lyratum as an active component. CONSTITUTION:Raw, dried or semi-dried leaves, stalks and/or whole grass of Solanum lyratum are leached at room temperature or extracted under heating with water, a water-soluble organic solvent (e.g. methanol, ethanol, etc.) or their mixture to obtain an extract of Solanum lyratum. The extract is used as an active component of the objective antitumor agent. The agent has excellent suppressing effect against various tumors, exhibits specific suppressing action even to Sarcoma 180 and JTC-26 and has extremely low toxicity. The extract is administered as it is or in the form of powder, granule, capsule or injection such as aqueous solution or suspension, etc., by oral administration or parenteral administration such as subcutaneous injection. The dose is 1g/ person daily for oral administration and 60mg/person daily for parenteral administration.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention is directed to Solanum lyra.
The present invention relates to an anti-W1 tumor agent containing an extract of tumThunb (Japanese and English, white haired wisteria) as an active ingredient.

[Prior art and its problems (Background of the invention)] Hiyodorino tsugo is a vine-like herb of the Solanaceae family that grows wild in the mountains and roadsides of Japan, India, mainland China, etc., and has been used as a folk medicine for a long time. It is known to have antipyretic, diuretic, tonic, stomachic, and chain 1rtt effects, but there is a general tendency to avoid using it because it is said to be toxic.
Furthermore, its efficacy, ingredients, toxicity, etc. have not yet been sufficiently investigated. In view of this current situation, the present inventors have determined that J & Min (two-1
After conducting a detailed study of its physiological effects, we discovered that the extract from water, etc. of Bulbul vulgaris has an antitumor effect, and unexpectedly does not have any toxicity to the extent that it is toxic, thus completing the present invention. I ended up doing it.

[Structure and effect of the invention]

The key point of the present invention, which was developed against the background of such circumstances, is to construct an antitumor agent containing Bulbul vulgare extract as an active ingredient.

As used herein, the term "Bulbul heptadum extract" refers to an extract obtained by extracting the Bulbul heptadium with water, a water-soluble organic solvent, or a mixture of water and a water-soluble solvent.

The extract used as an active ingredient in the present invention can be prepared by extracting any of fresh, dried or semi-dried chestnuts, stems, foliage, and leaves of Chrysanthemum vulgaris in water or a water-soluble organic solvent (e.g., tutanol, ethanol, etc.). −
After filtration, the extract obtained by leaching at room temperature with other alcohol M) or a mixture thereof at room temperature, or by heating and extracting, is usually spray-dried, freeze-dried, concentrated to dryness, etc. Obtained by drying using the drying method.

Note that it is also possible to use other known water-soluble organic solvents. The extract obtained in this way can be used in a variety of ways. It has an excellent suppressive effect on tumors. For example, as described below, Sarcoma 18
It showed excellent suppressive effect against JTC-26 (
It also shows a specific suppressive effect on cervical cancer cells (infected cervical cancer cells), and its toxicity is extremely low.

Oral administration or parenteral administration such as dermal injection is used for Bulbul aurifolia extract. For oral administration, the extract can be used as it is, but it can be prepared into powders or granules by adding excipients, binders, lubricants, shavings, etc., which are normally used for 91M, and following the usual method of manufacturing preparations. It is administered in the form of tablets, capsules, syrups, etc., and for parenteral administration, it is dissolved in an aqueous solution, PA fluid, an injection such as an oily or aqueous emulsion, or an aqueous liquid medium such as sterile water or physiological saline. Alternatively, it may be administered in the form of an injection prepared by suspending it. Hyodorinotsugo extract can be further purified by conventional methods such as dialysis and various chromatography for use.The dosage varies depending on the symptoms, age, and dosage form, but it is usually administered orally to adults. It is desirable that the amount be about 1 g per day for administration, and about 6012 per day for parenteral administration. It is not something that will be done.

Example 1 60A of dried leaves of Prunus japonicum, 16001 of water
' was added and extracted at 100℃ for 1 hour using an extract extractor, and the insoluble matter was filtered, and the resulting extract was concentrated under reduced pressure at 40℃, and further spray-dried to obtain 4 kg of yellowish brown dry extract powder.
I got it. The yield was 6.7%.

Example 2 All 945 & of Dried Red-bellied Bulbul was extracted with hot water at 100°C for 1 hour, and the insoluble matter was filtered and the extract was concentrated under vacuum at 40°C to obtain a soft extract (fluid extract) with a concentration of 12%.
I got 381. Dry extract powder amount per 1M1 of extract is 0
．． It was 281F/J11'.

Formulation Example 1 (Tablets) 2 kg of the dry extract powder prepared in Example 1 above was mixed with 4 kg of lactose and about 30 g of magnesium stearate, and this mixture was compressed into tablets with a diameter of 9 mm% using a single-shot tablet tablet.
Weight approx.: 113019 tablets! I left it behind. Take 3-4 tablets at a time, 3 times a day, depending on your symptoms.

Formulation Example 2 (Capsule) 186 mg of the dried extract powder produced in Example 1 was filled into hard capsules. This capsule is taken 1 to 2 capsules at a time, 103 times, depending on the symptoms.

Test Example 1 Suppressive effect on Sarcoma 180 using mice (! & Adjustment of ingredients) 700 g of air-dried stems of Sarcoma 1-Ugo were extracted with hot water at 60°C, and the extract was divided into solvents between water and 1-butanol. 51 to the aqueous layer and butanol (BuOH) Jf4.
1 and then both parts of the water layer ^-berlite X^
The mixture was fractionated into a methanol (MeOH) eluate and an aqueous eluate by column chromatography using D-2 (IIli product name).

(Sample administration and effect) Using the test solution obtained from the above fractionation, Sarcoma 180
The activity against was investigated. That is, Sarcoma 1
80 was implanted subcutaneously in the groin area, and the aqueous layer and l1uOl were added to the mouse.
When both lR4, aqueous eluate, and MeOH eluate were administered once a day at a ratio of 2000.3000.3000.11000 i/Ay body 1, for a total of 10 times, AI,
As shown in Table ■, proliferation was suppressed in all cases compared to the control group (non-administered group). In particular, the BuOH layer showed the strongest suppressive effect when the tumor weight i was about 173 in the control group.

Table 1 Table ■ Test Example 2 Suppressive effect on JTC-26 (injected cervical cancer-derived cells) using mice (sample: A9) Bulbul was extracted as shown in Figure 1 to obtain a sample. .

That is, 1.1A of air-dried bulbul stems were extracted three times with warm water at 60°C (repeatedly, the extract was heated at 60°C).
The mixture was concentrated under reduced pressure to obtain 133 g of suikokisu. After treating this with 400zN of ethyl acetate (^cOEt), the residue 12
4 g was suspended in water 31 and extracted by shaking with BuOH3t'. On the other hand, the stems after hot water extraction were extracted twice with 4eOH under heat, the extract was concentrated under reduced pressure, and MeOH extract 38
I got g.

In addition, when we searched each of Suiko Kiss, ^cOEt soluble part, BaO2 layer, water layer, and HeOH extract by thin layer chromatography, we obtained the chromatogram shown in MS2 diagram, and found that the ^cOEL soluble part contains fat, etc. However, it was found that sugar was mainly transferred to the aqueous layer.

Then, the BaO2 layer was concentrated under reduced pressure to obtain 51 g of a residue, and as shown in Figure 1, 5ephadex LH-20
Repeat column chromatography using (trade name), silica sol column chromatography, recrystallization, etc.
Steroid saponin 5L-0 (2), SL-[(1),
Sl.

-II (3) respectively as crystals 7.9.1.6.
3.4g was obtained. The yields based on dry raw materials are each 0.
It was 72.0.15.0.31%. Also Me01
1 extract was subjected to repeated silica sol column chromatography to obtain SL-1(1)3. h (0.36% based on dry raw material) was obtained.

(Administration and effects of !&) Steroid dosabonine 5L-0 (2) obtained by the above method
, 5L-1 (1), 5L-II (3) Niraite JTc
-26 (entered cervical cancer cells) was investigated.・H
EM Eagles 90%, Feta-ICalf
5eru+s (Product name: Microbio-logic
a1 Company 9I) 10% medium, and the compound was further adjusted to each concentration and injected. Five samples were prepared for each sample, and the additive-free samples were used as a control group at 37°C and 144°C.
The number of surviving cells was counted in an air stream at C02 for an hour, and the average value was compared with that of the control group to determine the growth inhibition rate. The results are shown in Table ■. From this result, 5L-1 (1)
Oh! (/5L-n (3) was found to have a strong inhibitory effect. 5L-II (3) does not show activity at low concentrations, but when the concentration is higher than 8.0 μ2/11, it rapidly increases to 100%. On the other hand, no activity was observed for the 70-stanol compound 5L-0(2).

As can be seen from the above experimental results examining the inhibitory effect on J'FC-28, subirostanol glycoside showed activity, but no activity was observed for 70 stanoyl glycoside.

Table ■ Acute toxicity test Example 1.
Acute toxicity tests were carried out by administering orally or intraperitoneally the Bulbul 1 Ugo extract obtained in step 2.

In the case of oral administration, a dose of 5.6 ml of the Bulbul notarabo extract obtained in Example 2 was administered to a group of 10 8-week-old male mice.
On g7, the body weight was adjusted to 11.2y/Ag body weight. LD after 72 hours. is 11,211/
It was more than b. In the case of intraperitoneal administration, the extract obtained in Example 1 was given at an extract concentration of 0.381/&
g.

0.95g/b, 1.9g/ktt, 3. By/kg
, 7. WLfIL was suspended in physiological saline as follows and administered to 10 4-week-old male mice per group.
was 10z17Ag. 72 o'clock rl
LDs after llJl. was 2.05y/Ay.

The LD of the hot water soluble fraction of dried leaves of Kuma bamboo is more than 10y/If when administered orally, and 2.20g/Ag (1
In 1975, Mr. Shibata of Sato Pharmaceutical University published an article in the Japanese Pharmacological Journal:!
ft: Pharmacological research on bear bamboo), but the LD of oral administration and intraperitoneal administration of Bulbul vulgaris extract is almost the same.

In addition, 4-week-old and 8-week-old male and female mice (8^LB/c,
nwcrj), 6-week-old female rats (Crj, Mista
r) Oh! /4 week old female guinea pig) (Crj, Har
subacute toxicity IJ using tley),! 9 (When administered for 8 weeks, the internal intake per animal body weight is approximately 38 g/Ag in terms of dry rice extract for mice and approximately 12.2 g/Ag for rats.
kg, guinea pigs are 14.8g/λ, mice are 55g/λ, rats are 16.7g after 13 weeks of administration.
g/kg, guinea pigs 24.4y/kg), but 8 weeks and 13 weeks Il! In the dormitory, no changes were observed due to the difference in period, the membrane condition was normal, and there were no significant differences in body weight change or food intake compared to the control group. In addition, no gender-related changes were observed in histopathological findings, protein metabolism, etc., and no significant differences were observed in blood cell counts, etc. compared to the control group.

Furthermore, histopathological observations of major organs etc. were normal in mice, rats, and guinea pigs, and overall there were no significant abnormalities.

From the above tests, it is clear that the toxicity of Bulbul Shogo extract is extremely low.

In addition, in the above-mentioned examples, etc., dried bulbul is used, but fresh or semi-ripened bulbul is used! , it is possible to use dried ones.

[Brief explanation of the drawing]

Figure 1 is a diagram of the fractionation process of the test sample of the present invention;
The figure is a thin layer chromatogram of Hyodorino lI+7 Gogo extract.

Claims (1)

[Claims] An anti-tumor agent containing bulbul funnel extract as an active ingredient.