JPS628415B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an antitumor agent containing saponin components present in medicinal ginseng as an active ingredient.

Medicinal ginseng, especially Araliaceae
Panax ginseng (Panax ginseng, C.A. Meyer, Panax ginseng C.
A. MEYER), also known as Panax ginseng, is widely known to have been used as a tonic, tonic, anti-inflammatory, diuretic, blood pressure lowering, and anti-diabetic drug since ancient times. Recently, research has been underway to investigate whether these medicinal effects may be due to the saponin components in Panax ginseng. However, it is completely unknown that this saponin component has antitumor and anticancer effects.

This invention was made based on the new finding that saponin components in medicinal ginseng unexpectedly have antitumor effects (including cancer).

According to one aspect of the present invention, there is provided an antitumor agent containing saponin components of medicinal ginseng as an active ingredient.

As the carrot containing the saponin component in this invention, Panax ginseng is the most preferred. Other related plants include Panax japonicus (Panax japonicus, Panax japonicus CA, Panax japonicus CA)
MEYER), American ginseng (Panax quinquefolium, Linnaeus, Panax quinquefolium)
LINNE), Panax pseudo-ginseng (Panax pseudo−)
ginseng WALICH or Panax notoginseng
BURKILL).

The saponin component of the present invention can be obtained by extracting, separating, and purifying the herbal medicine using the above carrot as a raw material, or by tissue culturing the above-mentioned carrot rhizome section, and then extracting, separating, and purifying it. can. In the present invention, the term "saponin component" refers to a mixture substantially consisting only of saponins obtained by these methods.

Saponin components can be obtained from the herbal medicine of carrots, for example, by the following method. That is, the carrots used as raw materials are degreased without being defatted, or after being defatted using a normal fat-soluble organic solvent, and the active ingredients are extracted using water, lower aliphatic alcohols, or hydrous lower aliphatic alcohols, and the extract is made into an extract. is evaporated and concentrated to obtain an extract. This is dissolved in n-butanol, water is added to the solution, shaken and left to stand to remove insoluble substances, and the n-butanol layer is evaporated to dryness. After dissolving the residue in a lower aliphatic alcohol, the resulting precipitate may be collected by stirring and pouring into ether (see Japanese Patent Publication No. 5016/1983).

The extract thus obtained contains substantially only saponin components and can be used as is as an active ingredient in the present invention.

The saponin component according to the present invention has some differences in the type and amount of the component depending on the type of medicinal ginseng used as raw material and the number of years of culturing, but the saponin component according to the following formulas (), () and () are used. It contains a compound. The overall properties of the saponin components are that they are all yellowish-white to colored powders with a bitter taste, easily soluble in water, methanol, and diluted methanol, soluble in ethanol, and insoluble in chloroform, ether, and carbon tetrachloride. . In addition, acid hydrolysis always yields glucose from the water-soluble portion, and panaxadiol (C ₃₀ H ₅₂ O ₃ , mp205°C) and/or panaxatriol (C ₃₀ H ₅₂ O _Four ,
mp238-239℃) is obtained.

In addition, to obtain saponin by tissue culture, for example, Knop culture solution (calcium sulfate 1000
mg/, potassium nitrate 250mg/, magnesium sulfate 250mg/, monobasic potassium phosphate 250mg/)
500ml, d'Heller mineral liquid 1ml/
, glucose 5%, vitamin B10 ^-6 g, biotin
A piece of carrot root tissue is placed in a medium containing 10 ^-6 g of kinetin, 10 ^-6 g of kinetin, and 1% agar, and cultured and grown at 26°C to obtain carrot callus. Next, this callus was grown in large quantities using the same medium,
A saponin component can be obtained by extracting and purifying this product in the same manner as the extraction method described above.

In addition, the saponin component in this invention contains at least one type of ginsenoside represented by the following formula () or (), and if necessary, β-D-
Further included is glucopyranosyloleanate-(3)-β-D-glucopyranosyl(1→2)-β-D-glucuronopyranoside.

formula(): In the formula, R ¹ is β-D-glucopyranosyl (1→
2) -β-D-glucopyranosyl group, R ² is β-D-glucopyranosyl (1→6)-β-D-
Glucopyranosyl group, α-L-arabinopyranosyl (1→6)-β-D-glucopyranosyl group, β
-D-xylopyranosyl (1-6)-β-D-glucopyranosyl group, α-L-arabinofuranosyl (1→6)-β-D-glucopyranosyl group, or -
Indicates a β-D-glucopyranosyl group.

formula(): In the formula, R ³ is α-L-rhamnopyranosyl (1→
2) -β-D-glucopyranosyl group, β-D-glucopyranosyl (1-2)-β-D-glucopyranosyl group, β-D-glucopyranosyl group or α
-L-rhamnopyranosyl (1→2)-β-D-glucopyranosyl group, and R ⁴ represents a hydrogen atom or a β-D-glucopyranosyl group.

formula(): In the formula, R ⁵ represents β-D-glucopyranosyl group, and R ⁶ represents β-D-glucopyranosyl (1-2)-
Indicates a β-D-glucuronopyranosyl group.

The saponins represented by formulas () and () belong to the dammarane glycosides of triterpenes. These saponins of formula () and formula () are currently known to be specifically contained only in medicinal ginseng, and are considered to constitute the main body of pharmacological activity as an antitumor agent according to the present invention. Conceivable.

Specific names of the compounds represented by formula () include 20S-protopanaxadiol-3-
[0-β-D-glucopyranosyl(1-2)-β-
D-glucopyranoside]-20-[0-β-D-glucopyranosyl (1→6)-β-D-glucopyranoside] (ginzenoside Rb ₁ ), 20S-protopanaxadiol-3-[0-β-D-glucopyranosyl (1→2)-β-D-glucopyranoside]-20
-[0-α-L-arabinopyranosyl (1→6)-
β-D-glucopyranoside] (ginzenoside
Rb ₂ ), 20S-protopanaxadiol-3-[0
-β-D-glucopyranosyl (1→2)-β-D
-glucopyranoside]-20-[0-α-L-arabinofuranosyl (1→6)-β-D-glucopyranoside] (ginzenoside Rc), 20S-protopanaxadiol-3-[0-β-D- Glucopyranosyl (1→2)-β-D-glucopyranoside]-20-
[0-β-D-xylopyranosyl (1→6)-β-
D-glucopyranoside] (ginsenoside Rb ₃ ) and 20S-protopanaxadiol-3-[0-
β-D-glucopyranosyl (1→2)-β-D-
glucopyranoside]-20-[0-β-D-glucopyranoside] (ginsenoside Rb).

What is the specific name of the compound represented by formula ()?
20S-protopanaxatriol-6-[0-α
-L-rhamnopyranosyl (1→2)-β-D-glucopyranoside] -20-0-β-D-glucopyranoside (ginzenoside Re), 20S-protopanaxatriol-6-0-β-D-glucopyranoside (1→ 2) -β-D-glucopyranoside (ginzenoside Rf), 20S-protopanaxatriol-6-,20-di-0-β-D-glucopyranoside (ginzenoside Rg ₁ ), 20S-protopanaxatriol-6-0 -α-L-rhamnopyranosyl (1→2)-β-D-glucopyranoside (ginzenoside Rg ₂ ) and 20S-protopanaxatriol-6-[0-β-D-glucopyranosyl (1→2)-β-D- Glucopyranoside〕-20-0
-β-D-glucopyranoside (ginzenoside 20
-Gluco-Rf).

In addition, Panax ginseng contains the aforementioned formula (),
In addition to saponins having the structural formulas () and (), saponins of undetermined structure called ginsenoside Ra, which is thought to have the skeleton of formula (), and ginsenoside Rh, which is thought to have the skeleton of formula (). These substances are also included in the saponin component of this invention (Chem Pharm,
Bull., 22(2) , 421-428 (1974) and Pharmaceutical Journal
94(2) , 252-260 (1974)).

The above-mentioned individual compounds can be prepared by mixing the saponin component obtained as described above with, for example, a chloroform/methanol/water system or n-butanol/acetic acid/
It can be obtained by separating and purifying each constituent saponin by silica gel column chromatography, high performance liquid chromatography, etc. using an aqueous developing solvent. However, from an economical point of view, it is preferable to use the saponins as a mixture rather than separating them into the individual constituent saponins.

Saponin components are extremely useful as antitumor agents for humans, have a wide range of application, and have almost no side effects.

The dosage of the antitumor agent in this invention varies depending on the disease state, but when administered internally to adults, the saponin component is 50 to 1000 mg per day, preferably
It can be effective by administering 100 to 300 mg in two to three doses. For external use, it is used in the form of a 1-10% hydrophilic ointment or hydrophobic ointment. Applicable ranges include stomach cancer, rectal cancer, breast cancer, uterine cancer, oral cavity cancer, esophageal cancer, bile cancer, bile duct cancer, biliary tract cancer, pancreatic cancer, kidney tumor, prostate cancer, subthyroid tumor, lung cancer, brain tumor, and liver cancer. It is effective against almost all tumors including cancer, such as tongue cancer, thymoma, skin cancer, and sarcoma.

The antitumor agent according to the present invention consists of the saponin component of the present invention alone, or the saponin component and a solid or liquid excipient. The administration method and dosage form are usually powders, tablets, emulsions, capsules, teas, granules, liquids (including spirits, tinctures, liquid extracts, syrups, etc.) and other oral medications. It has a shape. It may also be injected into the body in the form of injections or drops, or externally administered in the form of ointments, liquids, external powders, syrups, suppositories, sprays, nourishing enemas, emulsions, and the like. Solid or liquid excipients used here include:
Those known in the art are used. It may be desirable to formulate the compound to contain the required amount of the compound for a single dose, such as those described above.

Excipients in powders and other powders for internal use include lactose, starch, dextrin, calcium phosphate, calcium carbonate, synthetic and natural aluminum silicates, magnesium oxide, dry aluminum hydroxide, stearin, to name a few specific examples. Magnesium acid, sodium bicarbonate, dried yeast, etc., and for external powders, zinc oxide, talc, starch, kaolin, boric acid powder, zinc stearate, magnesium stearate, magnesium carbonate, precipitated calcium carbonate, hypogallic acid. Examples include bismuth and potassium aluminum sulfate powder. Excipients for liquid preparations include water, glycerin, propylene glycol, simple syrup, ethanol, fatty oils, ethylene glycol, polyethylene glycol, sorbitol, and the like. Furthermore, in the case of ointments, fats, fatty oils, lanolin, petrolatum, glycerin, beeswax, Japanese wax, paraffin, liquid paraffin, resins, higher alcohols, plastics, glycols, water,
Hydrophobic bases or hydrophilic bases (including emulsion bases, water-soluble bases, and suspension bases) prepared by combining surfactants and the like are used as excipients.

On the other hand, when the saponin component according to the present invention was administered intraperitoneally to mice, the toxicity was extremely low with an LD ₅₀ of 637 mg/Kg, and no hemolytic effect was observed.

Next, manufacturing examples and clinical trial examples according to this invention will be described.

Production example 10 kg of dried roots of Panax ginseng (4th grade) were cut into small pieces, heated and extracted three times for 3 hours each with 100 methanol, and the extracts were combined and concentrated to 10. The concentrated solution was gradually poured into 100% ether in small portions with stirring, and the precipitate was separated and dried until the ether odor disappeared. The product was dissolved with stirring on a steam bath three times for about 1 hour each with 10 parts of water-saturated n-butanol. The obtained solution is 3
Rinse three times with n-butanol saturated water to remove contaminating sugars and pigments by transferring them to the water.
The separated water-saturated n-butanol layer was distilled under reduced pressure at 80° C. or lower to dryness. The residue was dissolved in 3 parts of methanol and poured into 6 parts of ether with stirring.
After standing for one day, the precipitate was separated and dried under reduced pressure at 60° C. or lower to obtain 260 g of carrot saponin.

The following clinical trial examples were obtained using the carrot saponin thus produced.

Clinical trial case 1: A 62-year-old male was histopathologically diagnosed with gastric cancer, but he refused gastrectomy, so he did not undergo surgery and
200 mg of carrot saponin was administered in two doses on an empty stomach. At the start of medication, weight 36 kg, red blood cell count
4,150,000, white blood cell count 6,600, facial pallor, severe epigastric pain, and palpation revealed a mass. One month after starting the medication, the epigastric pain disappeared and the patient's weight increased to 41 kg. The patient's complexion improved, and gastric fluoroscopy revealed only a small tumor, which was diagnosed as an ulcer. Furthermore, after one month of administration, the patient's feeling of fatigue disappeared completely, his appetite increased, and he weighed 42.5 kg. At this time, endoscopy and biopsy revealed only an ulcer scar, and a biopsy of the scar area showed no malignant findings. After administration of ginseng saponin, there were no subjective symptoms or test findings that suggested acute or subacute toxicity due to ginseng saponin.
This is an example of an extremely effective effect after two months of administration.

Trial case 2: A 52-year-old female patient with gastric cancer accompanied by cancerous peritonitis. The cancer has spread throughout the stomach and has invaded the peritoneum, pancreas, tunica, and small intestine, making resection impossible. The patient complained of abdominal pain and a feeling of vomiting after eating, and was prone to constipation, with abdominal pain especially severe before defecation. Since the patient's face was pale and he was extremely weak, we administered 200 mg of carrot saponin per day in two divided doses on an empty stomach. Two weeks after administration, abdominal pain began to ease slightly, and an increase in intake was observed, albeit on a semi-liquid diet. Five weeks after administration, the body weight increased by 1 kg, and the abdominal pain was relieved to the extent that it was sometimes felt, but the pain before defecation was not alleviated, and no significant change was observed in the feeling of vomiting after eating.

Trial case 3 A 44-year-old female was diagnosed with breast cancer that had metastasized to the first lumbar vertebrae, and complained of severe lower back pain and difficulty walking. As carrot saponin therapy, 100 mg of carrot saponin per day was administered in two divided doses on an empty stomach. At the start of administration, the red blood cell count was 3.8 million, the white blood cell count was 6000, and the erythrocyte sedimentation rate was 20 for 1 hour and 46 for 2 hours. One week after taking the medication, the patient's lower back pain was slightly relieved and her complexion began to improve, and after five weeks, the pain completely disappeared and she was able to walk again. At this time, the number of red blood cells was 4 million, the number of white blood cells was 6400,
The erythrocyte sedimentation rate was 16 at 1 hour and 36 at 2 hours. Furthermore, X-ray examination findings of the bone metastasis site showed no progression of metastasis at all. After the administration of carrot saponin was started, an increase in appetite was observed, but no change in body weight was observed. Furthermore, during the 5-week administration period, no subjective or objective symptoms suggesting side effects caused by carrot saponin were observed.

Trial case 4: A 62-year-old male patient with rectal cancer with intra-abdominal metastasis.
A mass was palpated in the umbilical region, and the patient complained of lower back pain and pain in the anal region during defecation. Red blood cell count 415
The patient's white blood cell count was 7,300, and X-rays showed no metastasis to the lungs. As in Trial Examples 1 and 3, 100 mg of carrot saponin per day was administered in two divided doses on an empty stomach. Four weeks after administration, lower back pain began to ease,
My appetite also started to increase. However, the pain in the anal region during defecation did not subside. At this time, the number of red blood cells was 4.39 million,
White blood cell count was 8900 and an increase in lymphocytes was observed. From this time on, the dose was increased to 200mg per day (twice) and continuous administration was carried out for 4 weeks.As a result, the mass in the umbilicus shrank to the extent that it could be slightly palpated, and the lower back pain completely disappeared. Disappeared.

Claims (1)

[Scope of Claims] 1. An antitumor agent containing saponin components of medicinal ginseng as an active ingredient. 2. Medicinal carrots include Panax ginseng (Panatucus ginseng, C.A. Meyer), Horse chestnut ginseng (Panatchus japonicus, C.A. Meyer), American ginseng (Panatucus cinquefolium, Linnaeus), Panax ginseng (Panatucus pseudoginseng, C.A. Meyer), The antitumor agent according to claim 1, which is Panax or Panax notoginseng, Burkil). 3. The antitumor agent according to claim 1, wherein the medicinal ginseng is Panax ginseng. 4. The antitumor agent according to claim 2 or 3, wherein the saponin component of medicinal ginseng is an extract from ginseng according to claim 2 or 3.