WO2013146734A1 - 抗癌剤 - Google Patents

抗癌剤 Download PDF

Info

Publication number
WO2013146734A1
WO2013146734A1 PCT/JP2013/058692 JP2013058692W WO2013146734A1 WO 2013146734 A1 WO2013146734 A1 WO 2013146734A1 JP 2013058692 W JP2013058692 W JP 2013058692W WO 2013146734 A1 WO2013146734 A1 WO 2013146734A1
Authority
WO
WIPO (PCT)
Prior art keywords
burdock
arctigenin
arctiin
extract
anticancer agent
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2013/058692
Other languages
English (en)
French (fr)
Japanese (ja)
Inventor
江角　浩安
公史 池田
千香 三好
重利 門田
敏樹 大窪
与茂田　敏
布施　貴史
孝則 川島
殖幹 千葉
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kracie Pharma Ltd
National Cancer Center Japan
University of Toyama NUC
National Cancer Center Korea
Original Assignee
Kracie Pharma Ltd
National Cancer Center Japan
University of Toyama NUC
National Cancer Center Korea
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kracie Pharma Ltd, National Cancer Center Japan, University of Toyama NUC, National Cancer Center Korea filed Critical Kracie Pharma Ltd
Priority to JP2014507890A priority Critical patent/JP5707534B2/ja
Priority to CN201380016479.1A priority patent/CN104379143A/zh
Priority to EP13767892.6A priority patent/EP2832355A4/en
Priority to HK15103344.0A priority patent/HK1203811A1/xx
Priority to IN1973MUN2014 priority patent/IN2014MN01973A/en
Priority to KR1020147029903A priority patent/KR20140139073A/ko
Publication of WO2013146734A1 publication Critical patent/WO2013146734A1/ja
Priority to US14/494,637 priority patent/US9700572B2/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
    • A61K31/341—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide not condensed with another ring, e.g. ranitidine, furosemide, bufetolol, muscarine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to an anticancer agent containing arctigenin. More specifically, the present invention relates to an anticancer agent containing architigenin so that the daily dose is 100 mg or more.
  • Pancreatic cancer is one of the intractable cancers, and the 5-year survival rate of all patients is estimated to be 2-3%.
  • the number of patients who die from pancreatic cancer has increased rapidly by approximately 2.5 times over the last 20 years, with 2009 statistics showing 26,791 deaths from pancreatic cancer.
  • the number of occurrences and deaths is almost the same, accounting for 6% of cancer deaths in Japan, and the fifth in cancer deaths by region, after lung, stomach, large intestine, and liver.
  • Surgical excision is the only treatment that can be expected to be cured.
  • pancreatic cancer is found in many patients with advanced cancer (stage III + IV), it is actually possible to remove the pancreas. It is said to be about 10-20% of all cancer patients.
  • the median survival time for each stage was about 12-30 months for Stage I and II, 9-11 months for Stage III, and 5-6 months for Stage IV. It is thought that there is almost no possibility of doing.
  • pancreatic cancer-derived cells such as PANC-1, AsPC-1, BxPC-1 and KP-3 have been found to be highly resistant even under extreme nutrient starvation conditions.
  • PANC-1 pancreatic cancer-derived cells
  • AsPC-1 AsPC-1
  • BxPC-1 BxPC-1
  • KP-3 KP-3
  • Non-patent Document 1 When the pancreatic cancer cell line PANC-1 was used to screen for substances capable of canceling the viability of tumor cells in a low nutrient state, it was reported that archigenin is effective (Non-patent Document 1).
  • burdock is defined as the fruit of Burdock Arctium lappa Linne (Compositae) in JP15.
  • burdock is a herbal medicine prescribed for Ginkakusan, Gakufu-Dokuto, Shifufu-san, etc., and is classified as a component essence used exclusively as a medicine.
  • the burdock contains about 7% of arctiin, which is classified as a lignan glycoside, and about 0.6% of its aglycone, arctigenin. From these findings, it is expected that the burdock extract containing arctigenin can be used as an anticancer agent for treating cancer.
  • An object of the present invention is to provide a novel anticancer agent having an effect on cancer.
  • the currently known burdock has a low architigenin content of about 0.6%. Also, it is difficult to dissolve in water. For this reason, it has been extremely difficult to produce a burdock extract containing a high content of arctigenin by a conventionally used hot water extraction method.
  • argotigenin is a main active ingredient, and a gobo cow extract containing architigenin and archtiin at a certain content is particularly excellent in anticancer effect.
  • a production method that can control architigenin and archtiin so as to have a constant content is desired.
  • a method is desired that can produce a burdock extract that contains arctigenin and arctiin in a weight ratio of about 1: 1.
  • the present inventors have found that the raw ⁇ -glucosidase enzyme activity as a raw material, the particle size for cutting the burdock, and the enzyme conversion of archtiin to architigenin.
  • the present inventors have found a technique for adjusting the content of arctigenin and arctiin, and a technique for adjusting the content ratio of arctigenin and arcthiin by adjusting the temperature and the temperature at which arctigenin and arctiin are extracted from burdock.
  • the present inventors administered an agent prepared so that the daily dose of archigenin was 100 mg or more using the gobo-bovine extract obtained by these techniques, the tumor was treated with a tumor. While observing the reduction effect, a decrease in the tumor marker was confirmed.
  • the present invention has been completed based on this surprising finding.
  • the present invention provides an anticancer agent containing archigenin so that the daily dose is 100 mg or more.
  • the present invention further provides the above-mentioned anticancer agent further containing alktiin so that the daily dose is 100 mg or more.
  • the present invention also provides the above anticancer agent, wherein the above arcticenin and the above arctiin are derived from burdock.
  • the present invention also provides the above anticancer agent for treating pancreatic cancer that is unresponsive to treatment with gemcitabine.
  • the present invention also provides an anti-cancer stem cell agent containing archigenin.
  • the present invention also provides an anti-cancer stem cell agent containing archigenin so that the daily dose is 100 mg or more.
  • a novel anticancer agent having an effect on cancer can be provided.
  • the figure which shows the result of the thoracoabdominal-contrast-enhanced CT before the granule administration of one Example of this invention (A), 1 month after the start of administration (B), 2 months later (C) and 3 months later (D) .
  • the figure which shows the result of the thoracoabdominal-contrast-enhanced CT before the granule administration of one Example of this invention (A), 1 month after the start of administration (B), 2 months later (C) and 3 months later (D) .
  • the figure which shows the result of the thoracoabdominal-contrast-enhanced CT before the granule administration of one Example of this invention (A), 1 month after the start of administration (B), 2 months later (C) and 3 months later (D) .
  • the figure which shows the result of anti-tumor property evaluation in a tumor model animal (CAPAN-1 Xenografts).
  • the anticancer agent of the present invention contains archigenin as an active ingredient so that the daily dose is 100 mg or more.
  • Arctigenin may be derived from a plant containing arctigenin, and may be derived from, for example, burdock. That is, the anticancer agent of the present invention may contain, as an active ingredient, arctigenin contained in an extract from a plant, for example, a burdock extract obtained from a burdock.
  • the anticancer agent of the present invention may further contain alktiin as an active ingredient so that the daily dose is 100 mg or more.
  • Arctiin may be derived from a plant containing arctiin, and may be derived from, for example, burdock. That is, the anticancer agent of the present invention may contain, as an active ingredient, arctiin contained in an extract from a plant, for example, a burdock extract obtained from a burdock.
  • architigenin may be contained so that the daily dose is 100 mg or more, and archtiin is contained so that the daily dose is 100 mg or more. Also good.
  • Plants containing arctigenin and arctiin are not particularly limited.
  • burdock sprouts, leaves, rhizomes
  • safflowers cornflowers
  • American thistles scorpionfish (Gifts thistles)
  • cardons sandworms
  • Anurocoreamis Ayokorengyou, Ayakorengi
  • Forsythia syringae, sesame, maple convolvulus, cinchiku himehagi, dung beetle katsura, taikakazura, munintakekakazura, himetakekakazura, tokyo oleander, keteka quail, rhododendron, okterade, yamazakuru Including Mexican cypress and kaya.
  • burdock and forsythia are preferred because of their high arctigenin content.
  • a burdock extract obtained using a method for producing a burdock extract described later can be used. Therefore, productivity at the time of manufacture can be improved, and an anticancer agent can be prepared inexpensively and easily. Moreover, also when using plants other than burdock, it is possible to easily obtain an extract containing arctigenin and arctiin by utilizing the production method described later.
  • the anticancer agent of the present invention can further contain an optional component.
  • the anticancer agent of the present invention can be provided in a form containing a pharmaceutically acceptable base, carrier, excipient, disintegrant, lubricant, colorant and the like.
  • Examples of carriers and excipients used for anticancer agents include lactose, glucose, sucrose, mannitol, dextrin, potato starch, corn starch, calcium carbonate, calcium phosphate, calcium sulfate and crystalline cellulose.
  • binder examples include starch, gelatin, syrup, tragacanth gum, polyvinyl alcohol, polyvinyl ether, polyvinyl pyrrolidone, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, carboxymethyl cellulose, and the like.
  • disintegrant examples include starch, agar, gelatin powder, crystalline cellulose, calcium carbonate, sodium hydrogen carbonate, sodium alginate, sodium carboxymethylcellulose, and carboxymethylcellulose calcium.
  • examples of lubricants include magnesium stearate, hydrogenated vegetable oil, talc and macrogol.
  • the colorant any colorant allowed to be added to a pharmaceutical product can be used.
  • Anticancer agents may be sucrose, gelatin, purified shellac, gelatin, glycerin, sorbitol, ethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, cellulose phthalate acetate, hydroxypropylmethylcellulose phthalate, methyl methacrylate and It may be coated with one or more layers with an acrylic acid polymer or the like. Moreover, you may add a pH adjuster, a buffering agent, a stabilizer, a solubilizer, etc. as needed.
  • anticancer agent can be provided as a preparation of any form.
  • anticancer agents include tablets such as sugar-coated tablets, buccal tablets, coated tablets and chewable tablets, capsules including troches, pills, powders and soft capsules, granules, suspensions, emulsions and dry syrups as orally administered preparations It can be a liquid such as a syrup or elixir.
  • anti-cancer agents are for parenteral administration, such as intravenous injection, subcutaneous injection, intraperitoneal injection, intramuscular injection, transdermal administration, nasal administration, pulmonary administration, enteral administration, buccal administration, and transmucosal administration.
  • the formulation for administration of can be an injection, a transdermal absorption tape, an aerosol, a suppository and the like.
  • the extract powder since the extract powder has a peculiar gumminess, it can be used as a preparation for masking the extract powder or a film coating agent coated with a coating agent.
  • the extract powder obtained by the method for producing a burdock extract described later can be used as an anticancer agent as it is.
  • the anticancer agent of the present invention may be produced by mixing purified arctigenin and other components, or may be produced by using a burdock extract produced by the method described below.
  • the burdock extract is produced through a herbal medicine cutting process, an extraction process (enzyme conversion process and extraction process using an organic solvent), a solid-liquid separation process, a concentration process, and a drying process.
  • the burdock as a raw material is cut into a size suitable for extraction.
  • Herbal medicines that are raw materials have various sizes, shapes, and hardness such as various parts of plants, minerals, and animals, and cutting according to their characteristics is necessary.
  • the burdock can be cut using any means known to those skilled in the art. For example, a commercially available cutting machine can be used.
  • the activity of ⁇ -glucosidase which is an enzyme inherent in the burdock, can be measured in advance, and a burdock suitable for the production of the present invention can be selected.
  • p-nitrophenyl- ⁇ -D-glucopyranoside C 12 H 15 NO 8 : molecular weight 301.25) (manufactured by SIGMA-ALDRICH) is used as a substrate to measure ⁇ -glucosidase activity.
  • SIGMA-ALDRICH p-nitrophenyl- ⁇ -D-glucopyranoside
  • U unit
  • burdock cut to an arbitrary particle size can be used. It is considered that the smaller the particle size of the cut burdock, the more the enzyme conversion is promoted and the extract yield is increased. On the other hand, if the particle size is too small, the enzyme conversion may be too fast, making process control difficult, and hindering accurate solid-liquid separation in subsequent steps.
  • the particle size of the burdock passes through the entire 9.5 mm sieve, for example, from 60 to 60 on a 0.85 mm sieve. It is desirable to cut so that 100% is distributed, and more preferably 65 to 80% is distributed on a 0.85 mm sieve.
  • extraction process is the most important process in terms of quality in the crude drug extract powder manufacturing process. This extraction step determines the quality of the herbal extract powder.
  • extraction is performed in two stages, an enzyme conversion process and an extraction process using an organic solvent.
  • the enzyme conversion process is an important process in the manufacturing method of the burdock extract used for the anticancer agent of this invention.
  • the enzyme conversion step is a step of converting arctiin contained in burdock into argtigenin by ⁇ -glucosidase, which is an enzyme inherent in burdock.
  • the burdock cut material prepared in the above step is maintained at an appropriate temperature to cause ⁇ -glucosidase to act, thereby causing the reaction from arctiin to arctigenin.
  • an arbitrary solution such as water is added to the cut burdock and stirred at a temperature of about 30 ° C., so that the burdock can be maintained at an arbitrary temperature.
  • the cut burdock is kept at a temperature around 30 ° C., for example, a temperature between 20 and 50 ° C.
  • the holding time is not particularly limited as long as it is held at the above temperature, and for example, it can be held for about 30 minutes.
  • an appropriate amount of archtiin is enzymatically converted to archigenin regardless of the retention time, resulting in a burdock extract containing about 1: 1 of archigenin: arctiin (weight ratio) be able to.
  • the extraction step with an organic solvent is a step of extracting arctigenin and arctiin from burdock using any appropriate organic solvent. That is, it is a step of extracting a burdock extract by adding an appropriate solvent in a state where the content of archigenin is increased by the enzyme conversion step.
  • a suitable solvent is added to the burdock extract, and the burdock extract is extracted by heating and stirring for a suitable time.
  • the burdock extract can be extracted using any extraction method known to those skilled in the art, such as heating reflux, drip extraction, immersion extraction, or pressure extraction.
  • arctigenin is sparingly soluble in water
  • the yield of arctigenin can be improved by adding an organic solvent.
  • Any organic solvent can be used as the organic solvent.
  • alcohols such as methanol, ethanol and propanol, and acetone can be used.
  • ethanol it is preferable to use ethanol as the organic solvent in the method for producing a burdock extract used for the anticancer agent of the present invention.
  • the time for heating and stirring is not particularly limited as long as the heating and stirring is performed at the above-mentioned temperature.
  • the time for heating and stirring is not particularly limited as long as the heating and stirring is performed at the above-mentioned temperature.
  • the yield of arctigenin and arcthiin increases as the heating and stirring time increases. However, if the heating and stirring time is long, a lot of unnecessary oils and fats are dissolved, and the load of the concentration process is increased. Therefore, what is necessary is just to determine the time of heat stirring suitably according to a condition.
  • the yield of arctigenin and arctiin increases as the amount of ethanol increases, so that the solubility of arctigenin and arctiin increases.
  • the burdock extract can be simultaneously sterilized and sterilized by heating and stirring in this step.
  • Solid-liquid separation process is a step of separating the burdock that has been extracted from the extract.
  • Solid-liquid separation can be performed using any method known to those skilled in the art. Examples of the solid-liquid separation method include a filtration method, a sedimentation method, and a centrifugal separation method. Industrially, a centrifugal separation method is desirable.
  • the concentration step is a step of removing the solvent from the burdock extract prior to drying. Removal of the solvent from the burdock extract can be performed using any method known to those skilled in the art.
  • the extract from the burdock obtained by the above process is not exposed to a high temperature for a long time.
  • the concentration of the burdock extract can be concentrated to a concentration at which a burdock extract having a desired concentration can be obtained.
  • dextrin can be added to the burdock extract obtained in this concentration step in order to prevent arctigenin and arctiin from adhering to the production apparatus.
  • the amount of dextrin added is preferably about 15 to 30% with respect to the solid content of the concentrate, for example.
  • Drying process It is a step of finishing the burdock extract obtained by the above step into a powder form. Drying can be performed using any method known to those skilled in the art. For example, freeze drying and spray drying are known as drying methods. The former is generally used at the laboratory level and the latter is used at the mass production level.
  • This method for producing burdock extract must include a step of performing enzyme conversion at a temperature of 20 ° C. to 50 ° C., but does not need to include all of the other steps.
  • the above-described production process makes it possible to obtain a burdock extract with a high concentration of arctigenin at low cost and in a simple manner. Therefore, by using the burdock extract obtained by this method, the anticancer agent of the present invention can be produced inexpensively and easily.
  • the arguchigenin concentration of the burdock extract obtained by the above manufacturing process is high, the total amount of the anticancer agent per day can be reduced as compared with the case of using the conventional burdock extract. Therefore, the burden on the patient can be reduced.
  • the present invention also provides a method for producing the above burdock extract, wherein the burdock is cut to a particle size of 0.85 mm to 9.5 mm in the cutting step.
  • the present invention provides a method for producing the above burdock extract, wherein the enzyme activity of ⁇ -glucosidase inherent in burdock is 0.4 U or more in 1 g of burdock.
  • the present invention also provides a method for producing the above-mentioned burdock extract, which comprises a step of extracting an extract containing arctigenin and arctiin by adding an organic solvent after the step of enzymatic conversion.
  • the present invention also provides a method for producing the above burdock extract, wherein the organic solvent is ethanol.
  • the present invention also provides a method for producing the above burdock extract, wherein the extraction step is performed at about 80 ° C.
  • the present invention provides a burdock extract obtained by the above method, which contains archigenin / arctiin in a weight ratio of 0.7 to 1.3.
  • the present invention provides an anticancer agent containing a gobo-bovine extract containing arctigenin / arctiin in a weight ratio of 0.7 to 1.3 obtained by the above method.
  • an anticancer agent having an antitumor effect which contains archigenin so that the daily dose is 100 mg or more.
  • an anticancer agent containing architigenin and archtiin at archigenin / arctiin 0.7 to 1.3 (weight ratio).
  • the anticancer agent of the present invention can provide a high anticancer effect with very few side effects.
  • arctigenin has a killing effect on pancreatic and liver cancer stem cells. This suggests that arctigenin exerts an anti-cancer effect on solid tumors by acting and killing not only the tumor body but also cancer stem cells. Arctigenin killed not only pancreatic cancer but also cancer stem cells of other cancers, and was shown to have an anticancer effect. Accordingly, the present invention provides anticancer agents for treating various cancers.
  • the present invention also provides an anticancer stem cell agent containing archigenin.
  • the anticancer stem cell agent refers to a drug having an effect of killing cancer stem cells.
  • Cancer stem cells refer to cells having stem cell properties among cancer cells.
  • the anticancer stem cell agent of the present invention can have the same configuration as the anticancer agent of the present invention described above. That is, the anticancer stem cell agent may contain archigenin so that the daily dose is 100 mg or more. In addition, the anticancer stem cell agent may further contain archtiin, and alktiin may be contained so that the daily dose is 100 mg or more.
  • Arctigenin and arctiin may be derived from a plant containing arctigenin and arctiin, for example, may be derived from burdock.
  • the anticancer stem cell agent of the present invention can be suitably used as an anticancer agent because it can suppress the growth of cancer cells by killing the cancer stem cells.
  • a substrate solution water was added to 0.15 g of p-nitrophenyl- ⁇ -D-glucopyranoside and the volume was adjusted to 25 mL to prepare a 20 mmol / L p-nitrophenyl- ⁇ -D-glucopyranoside aqueous solution.
  • a reaction mixture was prepared by adding 0.5 mL of p-nitrophenyl- ⁇ -D-glucopyranoside aqueous solution, and preheated at 37 ° C. for about 5 minutes.
  • Test Example 1 Add 7 mL of water to 1 g of cut burdock with an enzyme activity of 0.12, 0.27, 0.40 U / g (samples 1 to 3), and react at each reaction temperature at enzyme reaction temperature of 15 ° C and 20 ° C. The reaction time was set at 30 minutes, ethanol was added after the reaction, extraction was performed at 80 ° C., archigenin and alktiin in the obtained extract were quantified, and the weight ratio of archigenin / arctiin was determined. The results are shown in Comparative Examples 1-2 and Example 1 in Table 1.
  • Test Example 2 Add 7 mL of water to 1 g of cut burdock with an enzyme activity of 4.03 U / g (sample 5), and react at each reaction temperature under enzyme reaction temperatures of 30 ° C, 40 ° C, 50 ° C, and 60 ° C. The time was set to 15 minutes and 30 minutes (only at 30 ° C. and 60 ° C.), and after the reaction, extraction was performed with ethanol.
  • Example 3 The results are shown in Example 3 in Table 1.
  • architigenin / arctiin (weight ratio) 0.7
  • archigenin / arctiin (weight ratio) 1.0
  • the enzyme reaction temperature is preferably less than 60 ° C.
  • Example 3 Add 7 mL of water to 1 g of cut burdock with an enzyme activity of 1.42 U / g (sample 4), set the reaction time to 10 minutes and 30 minutes under the temperature condition of the enzyme reaction temperature 25 ° C, and after the reaction with ethanol Extraction was performed, and architigenin and alktiin in the obtained extract were quantified to obtain an architigenin / arctiin weight ratio.
  • Example 10 (Example 10 granule containing burdock extract powder) (1) Burdock extract powder of Example 7 33.3% (2) Lactose 65.2% (3) Hydroxypropylcellulose 1.5% Total 100%
  • burdock extract powder contained in the granule used in this test example contains 59.4 mg of arctigenin and 68.5 mg of arcutin. That is, the daily dose of architigenin administered to the patient is 148.5 mg and the archtiin is 171.25 mg.
  • Figures 1-3 show thoracoabdominal CT images before (A), 1 month after the start of administration (B), 2 months (C), and 3 months (D) in this patient.
  • FIG. Based on these images, the tumor shrinkage effect was determined over time according to the new guideline (RECISTguideline (version 1.1) for determining the treatment effect of solid cancer. As a result, the results are shown by arrows in FIGS. 2 and 3. On the other hand, no serious side effects were observed in the patient after administration of the granule of Example 10.
  • RECISTguideline version 1.1
  • the anticancer agent of the present invention is effective against pancreatic cancer.
  • it has been shown to be effective against pancreatic cancer that is unresponsive to treatment with gemcitabine.
  • the fact that the composition of the present invention is effective against pancreatic cancer has not been known in the past and cannot be easily conceived without actually administering it to humans. It is.
  • the anticancer agent of the present invention can be expected to be effective not only for pancreatic cancer but also for poorly vascular, hypoxic and undernutrition cancer such as colon cancer.
  • Example 5 For 3 patients with pancreatic cancer who did not respond to gemcitabine therapy as pretreatment, 3 g of granule of Example 10 (containing 1 g of gobo cow extract powder) was orally administered once daily after breakfast did. That is, in this test example, the daily amount of arctigenin administered to the patient was 59.4 mg, and the amount of arctiin was 68.5 mg.
  • Example 12 Production of burdock extract by chopping, cold extraction, and ethanol addition
  • the mixture was filtered through 4 pieces of gauze (100 mesh mesh), washed with 0.5 L of 30% ethanol, and the combined extract (1.5 L) was lyophilized.
  • the tumor model animal is a donor mouse tumor mass obtained by seeding the nude mouse (BALB-cAJnu / nu; CLEA JAPAN), a human pancreatic cancer cell line, CAPAN-1 or PSN-1, under the back of the skin.
  • the tumor model animal was a donor mouse tumor mass obtained by seeding the nude mouse (BALB-cAJnu / nu; CLEA JAPAN), a human pancreatic cancer cell line, CAPAN-1 or PSN-1, under the back of the skin.
  • Arctigenin (AG), arctiin (A), and gobo cow extract were dissolved in DMSO at a concentration of 10 mg / ml with physiological saline, and 50 ⁇ g per mouse 5 times a week. Orally.
  • Anti-tumor properties were evaluated by measuring the size of the tumor mass under the back of the skin.
  • Another 3 out of 15 patients were administered 7.5 g of the granule of Example 10 (containing 2.5 g of gobo cow extract powder, ie, 148.5 mg of arctigenin and 171.25 mg of arctiin) at a time. Further, another 9 patients were administered 12 g of the granule of Example 10 (containing 4 g of gobo cow extract powder, ie, 237.6 mg of arctigenin and 274 mg of arctiin) at a time.
  • the frequency of dose-limiting toxicity was 0 among 15 target patients.
  • the major Grade 3 or higher adverse events in the Phase I study of argommein-containing granules for patients with pancreatic cancer were serious GGT elevation, hyperglycemia, ALP elevation, and blood bilirubin elevation. There were no adverse events.
  • Glucose-containing medium is 4.75 g Dulbecco's Modified Eagle Medium 2 (Nissui Pharmaceutical) dissolved in water, sterilized by adding 12.5 ml 1M HEPES pH7.4 (DOJINDO, 342-01375), then 18.5 ml 10% NAHCO 3 , 10ml L-glutamine (SIGMA), 5ml Anti-Anti (Life technologies), 5ml MEM NON-ESSENTIAL AMINO ACID SOLUTION (SIGMA), 50ml FETAL BOVINE SERUM (biowest) deactivated in a warm bath at 56 ° C for 30 minutes And finally made 500 ml.
  • the glucose inhibition medium was prepared by adding 2-Deoxy-Glucose (2-DG) (Tokyo Chemical Industry) with a final concentration of 20 mM to a glucose-containing medium.
  • 2-DG 2-Deoxy-Glucose
  • a medium containing 3 ⁇ M arctigenin was prepared by adding arctigenin (Krasie Pharmaceutical Co., Ltd.) having a final concentration of 3 ⁇ M to a glucose-containing medium or a glucose inhibition medium.
  • the FACS buffer was prepared by dissolving 10 g of Bovine serum, alcohol-free protein free (Wako Pure Chemical Industries, Ltd.) in 1 L of PBS (-), adding sodium azide at a final concentration of 0.1%, and sterilizing by filtration.
  • Fluorescently labeled antibodies for FACS analysis were CD44 (338803 or 338807, biolegend), CD24 (311117, biolegend), ESA (324205, biolegend), and c-Met (11-8858, e-bioscience). The dyeing process was performed according to the data sheet attached to the product.
  • pancreatic cancer cell PANC-1 (ATCC No. CRL-1469) was inoculated in a glucose-containing culture medium and incubated overnight, a glucose-containing medium, a glucose-inhibiting medium, a glucose-containing medium containing 3 ⁇ M arctigenin, and glucose containing 3 ⁇ M arctigenin Cultured for 24 hours in each of the inhibition media.
  • PI staining dead cell staining
  • cancer stem cell marker staining were performed according to a conventional method, and analyzed by flow cytometry (FACS).
  • FACS flow cytometry
  • FIG. 6 shows the results of staining with PI staining and cancer stem cell markers (triple positive for CD44 + , CD24 + and ESA + (CD326)).
  • PI staining the cell viability was 78.20% in the glucose-containing condition, 68.53% in the glucose-inhibiting condition, and 69.50% in the presence of 3 ⁇ M archigenin in the glucose-containing condition, compared to 3 ⁇ M in the glucose-inhibiting condition.
  • the survival rate was 35.71%.
  • the ratio of CD44 + ESA + CD24 + cells (survival count) indicating pancreatic cancer stem cells as a result of cancer stem cell marker staining was 4.41% (441 cells) in glucose-containing conditions, and in glucose-inhibiting conditions in all analyzed cells. 6.51% (651) and 5.01% (501) in the presence of 3 ⁇ M archigenin in the glucose-containing condition, compared with 0.98% (98) in the presence of 3 ⁇ M archigenin in the glucose-inhibiting condition. Thus, arctigenin has been shown to have the effect of killing pancreatic cancer stem cells in glucose starvation conditions.
  • FIG. 7 shows the results of PI staining and staining of cancer stem cell markers (CD44 + , c-Met High double positive).
  • cancer stem cell markers CD44 + , c-Met High double positive.
  • the cell viability was 84.70% in the glucose-containing condition, 88.80% in the glucose-inhibiting condition, and 83.30% in the presence of 3 ⁇ M archigenin in the glucose-containing condition, compared to 3 ⁇ M in the glucose-inhibiting condition.
  • the survival rate was 27.50%.
  • Glucose-containing medium is 4.75 g Dulbecco's Modified Eagle Medium 2 (Nissui Pharmaceutical) dissolved in water, sterilized by adding 12.5 ml 1M HEPES pH7.4 (DOJINDO, 342-01375), then 18.5 ml 10% NAHCO 3 , 10 ml L-glutamine (SIGMA), 5 ml Anti-Anti (Life technologies), 5 ml MEM NON-ESSENTIAL AMINO ACID SOLUTION (SIGMA) And finally made 500 ml.
  • the glucose inhibition medium was prepared by adding 2-Deoxy-Glucose (2-DG) (Tokyo Chemical Industry) having a final concentration of 15 mM to a glucose-containing medium.
  • 2-DG 2-Deoxy-Glucose
  • a medium containing 4 ⁇ M arctigenin was prepared by adding arctigenin (Krasie Pharmaceutical Co., Ltd.) having a final concentration of 4 ⁇ M to a glucose-containing medium or a glucose inhibition medium.
  • pancreatic cancer cell Capan-1 (ATCC No. HTB-79) was seeded in a glucose-containing culture medium and incubated overnight, a glucose-containing medium, a glucose-inhibiting medium, a glucose-containing medium containing 4 ⁇ M arctigenin, and glucose containing 4 ⁇ M arctigenin The cells were cultured in an inhibition medium or 7 ⁇ M cisplatin (CDDP: Wako Pure Chemical Industries, Ltd.) for 24 hours.
  • CDDP Wako Pure Chemical Industries, Ltd.
  • PI staining dead cell staining
  • cancer stem cell marker staining the markers analyzed by flow cytometry (FACS) include CD44, which has been reported as a stem cell marker for pancreatic cancer , CD24 and ESA (CD326) triple positives were used.
  • the results of PI staining and cancer stem cell marker staining are shown in FIG.
  • the cell viability was 67.57% in the glucose-containing condition, 79.10% in the glucose-inhibiting condition, and 72.80% in the presence of 4 ⁇ M archigenin in the glucose-containing condition, compared to 4 ⁇ M in the glucose-inhibiting condition.
  • the survival rate was 48.73% in the presence of arctigenin and 58.08% with cisplatin treatment.
  • the ratio of CD44 + ESA + CD24 + cells (number of survivors) indicating pancreatic cancer stem cells as a result of cancer stem cell marker staining was 0.87% (65 cells) in glucose-containing conditions, and glucose inhibition conditions 1.40% (105) and 1.04% (78) in the presence of 4 ⁇ M arctigenin in the glucose-containing condition, compared to 0.24% (18) in the presence of 4 ⁇ M arctigenin in the glucose-inhibiting condition, In cisplatin treatment, it was 1.17% (88).
  • arctigenin has a killing effect on the cancer stem cell-like population under nutrient starvation conditions close to the environment in which cancer cells are placed in the body.
  • Arctigenin also acts on cancer stem cells that have the ability to supply cancer cells and reconstruct tumor tissue, as well as to act on the tumor body to suppress the growth of cancer cells. It was strongly suggested that it also has a killing effect.
  • the granule containing octigenin in Test Example 7 showed very low toxicity.
  • Arctigenin has gained low toxicity because it has a killing effect under nutrient starvation conditions and can reduce the effects on normal cells by acting not only on cancer body cells but also on cancer stem cells The possibility is suggested. Therefore, the present invention can provide an anticancer agent having a high anticancer activity with few side effects.
  • the present invention can be suitably used as an anticancer agent, particularly a pancreatic cancer therapeutic agent.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
PCT/JP2013/058692 2012-03-26 2013-03-26 抗癌剤 Ceased WO2013146734A1 (ja)

Priority Applications (7)

Application Number Priority Date Filing Date Title
JP2014507890A JP5707534B2 (ja) 2012-03-26 2013-03-26 抗癌剤
CN201380016479.1A CN104379143A (zh) 2012-03-26 2013-03-26 抗癌剂
EP13767892.6A EP2832355A4 (en) 2012-03-26 2013-03-26 CANCER AGENT
HK15103344.0A HK1203811A1 (en) 2012-03-26 2013-03-26 Anticancer agent
IN1973MUN2014 IN2014MN01973A (enExample) 2012-03-26 2013-03-26
KR1020147029903A KR20140139073A (ko) 2012-03-26 2013-03-26 항암제
US14/494,637 US9700572B2 (en) 2012-03-26 2014-09-24 Anti-cancer agent

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2012-069964 2012-03-26
JP2012069964 2012-03-26

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US14/494,637 Continuation US9700572B2 (en) 2012-03-26 2014-09-24 Anti-cancer agent

Publications (1)

Publication Number Publication Date
WO2013146734A1 true WO2013146734A1 (ja) 2013-10-03

Family

ID=49259985

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2013/058692 Ceased WO2013146734A1 (ja) 2012-03-26 2013-03-26 抗癌剤

Country Status (8)

Country Link
US (1) US9700572B2 (enExample)
EP (1) EP2832355A4 (enExample)
JP (1) JP5707534B2 (enExample)
KR (1) KR20140139073A (enExample)
CN (1) CN104379143A (enExample)
HK (1) HK1203811A1 (enExample)
IN (1) IN2014MN01973A (enExample)
WO (1) WO2013146734A1 (enExample)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2014224085A (ja) * 2013-04-27 2014-12-04 クラシエ製薬株式会社 抗癌幹細胞剤
JP2016079163A (ja) * 2014-10-14 2016-05-16 医療環境テクノ株式会社 腫瘍を処置するための組成物およびその製造方法
CN105832719A (zh) * 2016-04-28 2016-08-10 上海中医药大学附属龙华医院 牛蒡苷元在调节pp2a 活性中的应用
WO2017061559A1 (ja) * 2015-10-09 2017-04-13 クラシエ製薬株式会社 血管壁強化剤、血管壁を強化する方法および血管網形成抑制剤
JP2017195882A (ja) * 2016-04-25 2017-11-02 クラシエホールディングス株式会社 未分化細胞除去剤
JP2021080223A (ja) * 2019-11-21 2021-05-27 クラシエホールディングス株式会社 レンギョウ葉の加工方法および加工レンギョウ葉

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109045010B (zh) * 2018-09-19 2021-03-30 淮海工学院 一种具有抗肿瘤作用的药物组合物及其应用
US11412028B2 (en) 2019-02-08 2022-08-09 Bluepipes, Inc. Online platform and a method for facilitating sharing of data between users

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002065298A (ja) 2000-08-30 2002-03-05 Japan Science & Technology Corp 薬剤成分のスクリーニング方法
WO2010109961A1 (ja) * 2009-03-27 2010-09-30 クラシエ製薬株式会社 アルクチゲニン高含有ゴボウシエキス及びその製造方法

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101036644A (zh) * 2006-03-13 2007-09-19 海南盛科天然药物研究院有限公司 含牛蒡子苷元的药用组合物及其制备方法
KR20080107794A (ko) * 2007-06-08 2008-12-11 경남대학교 산학협력단 우방자 추출물을 함유하는 항암 조성물
KR101538450B1 (ko) 2010-09-27 2015-07-22 구라시에제약 주식회사 악티게닌 함유 우방자 추출물 및 그 제조방법
CN102440986B (zh) * 2010-10-08 2014-12-03 鲁南制药集团股份有限公司 牛蒡子苷元在制备防治辐射或化学品引起的骨髓抑制的药物中的用途

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002065298A (ja) 2000-08-30 2002-03-05 Japan Science & Technology Corp 薬剤成分のスクリーニング方法
WO2010109961A1 (ja) * 2009-03-27 2010-09-30 クラシエ製薬株式会社 アルクチゲニン高含有ゴボウシエキス及びその製造方法

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
AWALE,S. ET AL.: "Identification of arctigenin as an antitumor agent having the ability to eliminate the tolerance of cancer cells to nutrient starvation", CANCER RES., vol. 66, no. 3, 2006, pages 1751 - 1757, XP009179519 *
BOECK,S. ET AL.: "Second-line chemotherapy with pemetrexed after gemcitabine failure in patients with advanced pancreatic cancer: a multicenter phase II trial", ANN. ONCOL., vol. 18, no. 4, 2007, pages 745 - 751, XP055170329 *
LI,Y. ET AL.: "Implications of cancer stem cell theory for cancer chemoprevention by natural dietary compounds", J. NUTR. BIOCHEM., vol. 22, no. 9, 2011, pages 799 - 806, XP028259393 *
MUELLER,M.T. ET AL.: "Combined targeted treatment to eliminate tumorigenic cancer stem cells in human pancreatic cancer", GASTROENTEROLOGY, vol. 137, no. 3, 2009, pages 1102 - 1113, XP026793002 *
S. AWALE; J. LU; S. K. KALAUNI; Y. KURASHIMA; Y. TEZUKA; S. KADOTA; H. ESUMI, CANCER RES., vol. 66, no. 3, 2006, pages 1751 - 1757
See also references of EP2832355A4 *
YOO,J.H. ET AL.: "Lignans inhibit cell growth via regulation of Wnt/beta-catenin signaling", FOOD CHEM. TOXICOL., vol. 48, no. 8-9, 2010, pages 2247 - 2252, XP027166430 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2014224085A (ja) * 2013-04-27 2014-12-04 クラシエ製薬株式会社 抗癌幹細胞剤
JP2016079163A (ja) * 2014-10-14 2016-05-16 医療環境テクノ株式会社 腫瘍を処置するための組成物およびその製造方法
WO2017061559A1 (ja) * 2015-10-09 2017-04-13 クラシエ製薬株式会社 血管壁強化剤、血管壁を強化する方法および血管網形成抑制剤
JPWO2017061559A1 (ja) * 2015-10-09 2018-09-27 クラシエ製薬株式会社 血管壁強化剤、血管壁を強化する方法および血管網形成抑制剤
JP2017195882A (ja) * 2016-04-25 2017-11-02 クラシエホールディングス株式会社 未分化細胞除去剤
CN105832719A (zh) * 2016-04-28 2016-08-10 上海中医药大学附属龙华医院 牛蒡苷元在调节pp2a 活性中的应用
JP2021080223A (ja) * 2019-11-21 2021-05-27 クラシエホールディングス株式会社 レンギョウ葉の加工方法および加工レンギョウ葉
JP7526562B2 (ja) 2019-11-21 2024-08-01 クラシエ株式会社 レンギョウ葉の加工方法および加工レンギョウ葉

Also Published As

Publication number Publication date
HK1203811A1 (en) 2015-11-06
IN2014MN01973A (enExample) 2015-07-10
US20150196576A1 (en) 2015-07-16
JPWO2013146734A1 (ja) 2015-12-14
US9700572B2 (en) 2017-07-11
JP5707534B2 (ja) 2015-04-30
EP2832355A1 (en) 2015-02-04
CN104379143A (zh) 2015-02-25
KR20140139073A (ko) 2014-12-04
EP2832355A4 (en) 2015-12-23

Similar Documents

Publication Publication Date Title
JP5707534B2 (ja) 抗癌剤
CN114940487A (zh) 一种人参碳点及其制备方法和用途
KR101670590B1 (ko) Ｅｒｋ 신호활성 억제제를 유효성분으로 함유하는 암 줄기세포 성장 억제용 조성물
JP5696191B2 (ja) 抗癌幹細胞剤
KR20200143668A (ko) 구강암을 치료하기 위한 약제를 제조하기 위한 진세노사이드 m1의 용도
JP6234553B2 (ja) 抗癌剤および副作用軽減剤
CN101468061B (zh) 北青龙衣提取物、制备方法及制药用途
JP5946558B2 (ja) Ｐｓｆ１遺伝子発現抑制剤
TWI542347B (zh) The use of arctigenin for the preparation of anticancer agents
CN106309758B (zh) 一种抗胃肠癌的药物组合物
CN102477039B (zh) 瓜馥木内酰胺碱化合物的制备方法及其在制备抗肿瘤药物中的应用
US12144821B1 (en) Selective 3-prenyl 5-methyl ether, 7-o-rhamnoside flavone for the treatment of hepatic cancer
JP5441023B2 (ja) 腫瘍を処置するための組成物およびその製造方法
TWI515002B (zh) 用以預防及／或治療肝纖維化的方法及醫藥組合物
CN101690738A (zh) 牛筋果及其提取物在制备抗肿瘤药物中的新用途
TWI661831B (zh) 使用台灣白鳳菜之經水萃取的產物來治療肝細胞癌以及抑制肝細胞癌幹細胞的生長
CN109602775B (zh) 乳苣醇提物在制备抗乳腺癌药物中的用途
KR20040036092A (ko) 진세노사이드 Rh2 및 Rg3 항암 조성물
TWI235664B (en) Extract from the leaves of Toona sinensis Roem., and the preparation process and uses thereof
JP2017160129A (ja) 抗悪性腫瘍組成物
CN108096291B (zh) 一种杉科植物挥发油制备的注射液及其应用
KR20240041441A (ko) 제주 황칠나무 유래 화합물을 포함하는 유방암 예방 또는 치료용 약학 조성물
KR20250012943A (ko) 참좁쌀풀 추출물을 포함하는 간암 전이 억제용 조성물
CN102824398B (zh) 紫景天在制备治疗肿瘤药物中的应用
JP2008044905A (ja) 固形腫瘍を処置するための組成物およびその利用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13767892

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2014507890

Country of ref document: JP

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 20147029903

Country of ref document: KR

Kind code of ref document: A

REEP Request for entry into the european phase

Ref document number: 2013767892

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2013767892

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: DE