WO2015182865A1 - Pharmaceutical composition exhibiting effects of preventing decrease in white blood cells and restoring white blood cells, containing angelica gigas nakai extract as active ingredient - Google Patents

Abstract

The present invention relates to an Angelica gigas Nakai extract and, more specifically, to an Angelica gigas Nakai extract having effects of preventing a decrease in the number of white blood cells and restoring the number of white blood cells, containing decursin and decursinol angelate as active ingredients. Preferably, the Angelica gigas Nakai extract of the present invention contains 70 wt% or more of decursin and decursinol angelate.

Description

A pharmaceutical composition effective in preventing leukocyte reduction and restoring leukocytes using the extract of Angelica gigas

TECHNICAL FIELD The present invention relates to extracts of Angelica gigas, and more particularly, to extracts of Angelica gigas, which are mainly composed of decusin and decosinol angelate, which are effective in preventing the reduction of leukocytes and restoring leukocytes.

Chemotherapy (chemotherapy), which is a method of removing cancer cells generated in the body, is a systemic treatment method that works on cancer cells spread throughout the body by treating cancer using drugs, that is, anticancer drugs. Since anticancer drugs are designed to remove rapidly growing cancer cells, normal cells are also affected. As a result, after chemotherapy, there are side effects such as anemia, white blood cell and platelet count, mouth loosening, nausea, vomiting, diarrhea, hair loss, and reproductive dysfunction. In the case of chemotherapy, the patient is allowed to rest for an average of two to three weeks to wait for normal cells to recover. Then, the next treatment is performed.

In this case, the level of white blood cells dropped by chemotherapy often does not recover. In this case, granulocyte colony stimulating factor (G-CSF) is administered to increase leukocytes to reach a certain level. It was. However, the administration of G-CSF can cause cold body pain, headache (20%), joint pain (15%), high fever (12%), and seriously cause side effects such as shortness of breath and temporary shock. Limited dose only if none. In addition, despite the administration of G-CSF, white blood cell counts do not recover and chemotherapy cannot be continued. Therefore, there is an urgent need for the development of effective and no side effect leukocyte reduction and recovery agents or substances.

Decusin is a natural component of the pyranocoumarin family first isolated from ether extracts of Angelica decursiva Fr. et Sav. In Japan and has a molecular weight of 328.35 and a molecular formula of C ₁₉ H ₂₀ O ₅ (Tetrahedron Letters No. 14, 1461-1465, 1966). In particular, Angelica gigas, Angelica gigas Nakai, contains a large amount of decusin (J. Pharm. Soc. Korea 11, 22-26, 1967) and J. Pharm. Soc. Korea, 13, 47-50, 1969) and a large amount of decusin has also been isolated from the fruit of the oil sprouts (Peucedanum terebinthaceum Fischeret Turcz.). In addition, the inventors have found that a large amount of decusin is also contained in Angelica gigas Jiri, a product of Jiri (Korean Patent Publication No. 10-2009-0078267).

Decasinol angelate is a structural isomer having the same molecular weight and the same chemical formula as decusin, and the molecular formula below shows the structure of decusin and decosinol angelate.

[Revisions under Rule 91 19.05.2015]
Formula 1

[Revisions under Rule 91 19.05.2015]
Formula 2

Decusin and decosinol angelate can be extracted from Korean Angelica gigas, Trigella giorum, Body Sprouts, Oil Sprouts, and other plants, or produced using tissue culture techniques and organic synthesis techniques.

A real donkey is a Korean donkey, called a native donkey or a native donkey. It refers to a donkey native to Korea. It has reddish color and leaves that are large and round, while donkeys of other countries are white flowers. The range of its use has been used as a herbal medicine as a blood donor, and recent studies have reported that decursin, an indicator of Korean donkey, and dekerusin, a major component, improve blood flow and anti-helicobacter.

As for the refining method, since the Angelica gigas has a very high content of decansin and decosinol angelate in the dried matter (about 9% of the dry matter) and can be used without purification, it can cause hypertension. Newly established 'Method for Extracting Decusin and Decusinol Angelate from Angelica Angelica' and Registered it as Korean Patent (Korean Patent Publication No. 10-0509843) and also contains 95% or more of the main ingredient The extraction method of the extract and the antioxidant effect of the extract using it was studied and registered as Korean Patent No. 10-0893779 (name of the invention: extraction method of true Angelica extract with harmful active oxygen scavenging effect), 'geological metabolism' About the Angelica and Tricuspid Angelica Extracts Using Decusin and Decusinol Angelate as Effective Ingredients and Their Extraction Methods It has been registered in the Republic of Korea (Korea Patent Publication No. 10-0995891).

In addition, the inventor has registered an invention patent for the increase of sperm number of the Angelica gigas extract, which contains decursin and decursinol angelate as main ingredients (Korean Patent Registration No. 10-1144715), Single and repeated toxicity of this tablet (Molecular & Cellular Toxicology, 5 (2) 153-159), Genotoxicity (Molecular & Cellular Toxicology 5 (1), 83-87), Pharmacokinetics (absorption, distribution, metabolism, excretion) (JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY 19 (12), 1569-1572), and published research results on standardization (Korean Journal of Food and Nutrition Science, 38 (5), 653 ~). 656).

The inventors of the present invention, while conducting a multi-faceted study on the efficacy of the Angelica Angelica extract as described above, while the Angelica Angelica extract has an excellent effect on the reduction and recovery of leukocytes due to the administration of anticancer drugs, compared to the conventional drugs such as G-CSF It has been found to be effective and free of side effects, which has led to the completion of the present invention.

The present invention has been invented to solve the above-mentioned conventional problems, and provides a pharmaceutical composition comprising deucin and decosinol angelate as an active ingredient extracted from true donkey as a leukocyte reduction preventing and restoring substance which is effective and has no side effects. It aims to do it.

The present invention for achieving the above object is implemented by the following solving means.

The present invention provides a pharmaceutical composition useful for the prevention of leukocyte reduction and leukocyte recovery using the decansin and decosinol angelate extracted from Angelica gigas.

In another aspect, the present invention provides a pharmaceutical composition useful in preventing leukocyte reduction and leukocyte recovery, which comprises a real Angelica extract containing more than 70% by weight of deckerin and decusin angelate.

In addition, the dosage of the decosin and the decosinol angelate is characterized in that 0.5 mg / kg to 10 mg / kg, preferably the dosage is 0.5 mg / kg to 2 mg / kg .

The present invention has the following effects by the above configuration.

The present Angelica extract as an active ingredient of decosin and decosinol angelate provided by the present invention has an excellent effect on preventing the reduction of leukocyte levels and restoring the reduced leukocyte levels after chemotherapy as shown in clinical and animal experiments. Has

1 is a photograph of granules prepared using the extract of Angelica gigas extract in an airtight container. The purified product per gram of granules contains 166.7 mg (deckerin and decosinol angelate content is 120 mg, 72% pure).

Figure 2 is a diagram showing the results of experiments in a manner to reduce the leukocyte count by administering benzene to the blood vessels to prevent the reduction and recovery of leukocyte count in the blood.

Figure 3 is a diagram showing the results of experiments in a manner to reduce the leukocyte count by oral administration of benzene to the leukocyte reduction and recovery effect of the present invention in the blood.

The applicant will be described below in detail the solution of the present invention. Details of known technologies that may unnecessarily obscure the subject matter of the present invention will be omitted and related embodiments will be described in detail with reference to examples. The following examples illustrate the invention in more detail, but do not limit the scope of the invention.

The present invention relates to a pharmaceutical composition useful for the prevention of leukocyte reduction and leukocyte recovery using the decancin and decosinol angelate extracted from Angelica gigas.

The content of this embodiment is the decansin and deckersinol angelate extracted by the method using ethanol extracted by the method according to the Republic of Korea Patent Registration No. 10-0893779 registered by the present inventors, specifically, the decansin and Decker which are the main components of the extract of Angelica gigas This is the result of studying synol angelate. If the content of the main components of deckercin and decosinol angelate is low and the impurities are high, there is a side effect of blood pressure increase, and according to the experiment by purity, the extract having purity from 70% to 98% of purity without affecting blood pressure Was used.

The pharmaceutical composition of the present invention is prepared according to a method that can be easily carried out by a person skilled in the art, and sucrose, lactose mannose, dextrose, fructose, starch commonly used in the preparation. , Algin, gelatin, cellulose, etc., and polyhydric alcohols such as sorbitol, mannitol, xylitol, lactitol, ribitol, and the like to prevent agglomeration when preparing liquids, and surfactants such as polysorbitan and triton X-100. It includes, and may include acacia rubber, pectin, xanthan gum, agar and the like as a thickener. In addition, stabilizers, preservatives, flavors, sweeteners, lubricants and the like can be further added,

Citric acid, succinic acid, tartaric acid, succinic acid, malic acid, boric acid, acetic acid, hydrochloric acid, ascorbic acid, etc. may be used as acidity regulators to prevent degradation.

In the present invention, the preparation may be administered in oral and non-oral preparations, and oral preparations may be administered in tablets, capsules, solutions, granules, powders and the like. The dosage of the formulation can be from 0.5 mg / kg to 10 mg / kg.

The following examples are to confirm the effect on the prevention and recovery of the reduction of white blood cell count using the Angelica Angelica extract containing more than 70% by weight of deckerin and decusin angelate.

Angelica Angelica Extract Tablets

As a method of separating and concentrating Decusin and Decusinol Angelate from Angelica gigas, it was carried out using Korean Angelica gigas, Korean Angelica gigas. Originating in China, originating in Japan, and originating in Vietnam are of low content and are not economically valuable.

For the food and medicinal purposes, the extraction solvents were ethanol and ethanol, and purified water was used. Grind the raw Angelica Angelica to 40 mesh (40 mesh) or less, dry it to 5% or less of water content, and add 2 to 4 times the amount of Angelica Angelica pulverized with medicinal alcohol or alcohol (hereinafter referred to as' ethanol). More than 12 hours were extracted sufficiently and this was quantified by filtration. In this case, the content of decusin and decosinol angelate in the concentrate was 33%.

The above concentrate was evaporated to dryness to obtain a highly viscous material. One liter of ethanol per 1 kg of this substance was dissolved in the main component, and the resultant precipitate was centrifuged to remove the resulting precipitate for 10 hours at minus 20 degrees to obtain a supernatant. The supernatant was evaporated to dryness to give a concentrate.

The above concentrate was eluted with 50 liters of 60% ethanol (diluted ethanol). In this case, the concentrations of Decusin and Decusinol Angelate in the concentrate were 330 grams, and the amount of the main component that can be dissolved in 50 liters of 60% ethanol was also used.

After elution, centrifugation gave only a 60% ethanol layer. This was evaporated to dryness to give a concentrate. This was dissolved in 99% alcohol and the supernatant was taken to give a final concentrate.

Representative results of liquid chromatography (HPLC) showed that the content was 54.52% deckerin, 41.26% decosinol angelate, 0.299% decosinol and 96% of the total content.

Granule manufacture

12.5% of Angelica gigas extract was weighed and dissolved in 10-fold alcoholic alcohol. Sucralose 0.0522%, citric acid 1.11%, magnesium stearate 1%, silicon dioxide 2%, mannitol 38.75%, corn starch 44.59% of the mixture was mixed and added to the Angelica extract refined ethanol solution and kneaded and 20 mesh through an automatic granulator Prepared to a particle size of. The prepared granules were dried at 80 ° C. for at least 4 hours and stored in an airtight container (see FIG. 1, and FIG. 1 shows granules prepared using the extract of Angelica gigas). The tablets per gram of granules were 120 contains mg).

Dosing Results for Female Cancer Patients

The patient of Example 3 is a female woman with colorectal cancer who lives in Busan, South Korea. She was in a serious condition that was unable to detect leukocytes due to three cancer treatments while being admitted to Seoul National University Hospital. It was a patient. The female patient received G-CSF for leukocytosis, but the leukocyte count did not recover and showed 0 cell / ul. The female patient was then administered granules of Angelica gigas extract for 3 weeks. One gram of granules (120 mg as extract tablets) was administered once daily and showed significant efficacy of raising leukocyte levels to 1800 cells / ul 3 weeks after dosing. Afterwards, he was re-admitted to receive the 4th and 5th anticancer drugs. At this time, the granules of True Angelica extract were administered once a day. The leukocyte count was 2,800 cells / ul after the fifth cancer treatment.

Dosing Results for Male Patients (Clinical Results of Patients Formulated with Granulated Angelica Angelica Extract)

The patient of Example 4 is a man in his 50's who is a lymphatic patient living in Busan, South Korea. During the treatment of cancer, he received a white blood cell count of 600 cells / ul. It was a patient. The male patient was asked to take granules of Angelica gigas extract after the first anticancer drug for leukocyte increase. One gram of granules (120 mg as extract tablets) was taken once a day and showed significant efficacy of increasing white blood cell count to 1100 cells / ul after 3 weeks of administration. Since then, 1 g of granulated Angelica extract was administered once daily during chemotherapy, and the leukocyte count before the third treatment was 1,600 cells / ul and before the fourth treatment, the level was over 7,000 cells / ul. About one week after the fourth treatment, the male patient showed leukocyte counts of 10,000 cells / ul and the number of leukocytes after chemotherapy up to 6 was 15,000 cells / ul. In addition, a liver and kidney function test was performed on this patient at Pusan National University Hospital. Blood tests and biochemical tests showed no functional impairment.

Preventing Leukocyte Reduction in Animals

1) Preparation of experimental animals

SD rats (SD white rats) were purchased at about 200 grams, adjusted for one week, and turned off and on every 12 hours. Breeding using a sterile animal testing apparatus (SPF chamber, Oriental Signs), the humidity was maintained at 50 ± 5%, temperature 24 ℃ ~ 26 ℃. When the animals were in stock, they were used for experiments after 7 days of adaptation.

In the IV administration of benzene, 4 SD rats were used per group. In the oral administration of benzene, three animals were used in the normal group and the control group (benzene administration group), respectively, and five in the oral administration group.

2) Preparation of Suspending Agent

A certain amount of Angelica gigas extract, including the main components decosin and decosinol angelate, was taken and dissolved in ethanol. In addition, xylitol, sucralose, citric acid, D-sorbitol, and Tween-80 were dissolved in water to prepare a solution. Each concentration was 20% xylitol, 0.02% sucralose, 0.025% citric acid, 10% D-sorbitol, 0.05% Tween-80, and 1% ethanol.

In the specific manufacturing method, a tungsten was added to the ethanol solution in which the main component was dissolved, and then uniformly melted and mixed with the aqueous solution in which the remaining substances were dissolved. After mixing, the suspension was slowly prepared while stirring to uniformly distribute the precipitated main component. This is what we call "Agnex suspension."

3) Experimental Method by Intravenous Administration of Benzene

Benzene was administered intravenously and 0.2 ml / kg was administered for 3 weeks at 2 days intervals. Angelica Angelica extract was administered orally at a dose of 2 mg / kg before and after benzene administration.

The control group was a free diet after benzene administration, and the positive control group was 0.2 ml / kg benzene administered intravenously for three weeks at two-day intervals.

The experimental and preventive groups received oral administration of Agnex suspension at 2 mg / kg for 7 days, followed by benzene (0.2 ml / kg intravenously every two days) and Agnex suspension (orally daily). Group administered for 3 weeks,

Normal group was a free diet group.

Recovery control group is a group that maintains a free diet of white rats with reduced white blood cells by intravenously administering 0.2 ml / kg of benzene for 3 weeks at two-day intervals.

The recovery group is an oral administration of an Agnex suspension for 7 days to white rats that have reduced white blood cells by intravenously administering 0.2 ml / kg of benzene for 3 weeks at intervals of 2 days.

4) Experimental method by oral administration of benzene

Benzene was orally administered and 200 mg / kg was administered once daily for two weeks. Benzene was uniformly mixed with the same amount of corn oil at the time of administration. Angelica gigas extract (Agnex, Agnex) was mixed with benzene at a dose of 10 mg / kg was administered once a day, two weeks.

The control group is a group of natural diet after oral administration of benzene,

Positive control group received oral Benzene 200 mg / kg for 2 weeks daily

The oral administration group was an oral administration of Agnex suspension at 10 mg / kg dose for 2 weeks with benzene (200 mg / kg) simultaneously.

5) Experiment result

(1) Experimental Results by Intravenous Administration of Benzene

White blood cell count,

The control group (6,500 cells / ul) had a decrease of about 32% compared to the normal group (9,500 cells / ul).

The experimental and preventive groups (10,000 cells / ul) were found to be slightly higher than the normal group, increasing by 5.2%.

In the recovery group, the recovery control group (6,000 cells / ul) decreased about 8% more than the control group, while the recovery group (8,700 cells / ul) recovered to 91.6% of the normal group. This shows that the recovery experimental group showed a 45% increase in white blood cell count compared to the recovery control group (see FIG. 1).

(2) Experimental result by oral administration of benzene

White blood cell count,

The control group (3,660 cells / ul) had a 56% reduction compared to the normal group (6,460 cells / ul).

The experimental group (4,610 cells / ul) was restored to 71% level compared to the normal group and showed a 26% increase compared to the control group (see Figure 3).

The present Angelica extract as an active ingredient of decosin and decosinol angelate provided by the present invention has an excellent effect on preventing the reduction of leukocyte levels and restoring the reduced leukocyte levels after chemotherapy as shown in clinical and animal experiments. Has

Claims (4)

1. A pharmaceutical composition useful for the prevention of leukocyte reduction and leukocyte recovery using the decancin and decusinol angelate extracted from Angelica gigas.
2. A pharmaceutical composition useful for preventing leukocyte reduction and restoring leukocytes, the extract comprising Angelica gigas extract containing at least 70% by weight of decusin and decusin angelate.
3. The method according to claim 1 or 2,

   The pharmaceutical composition useful for preventing leukocyte reduction and leukocyte recovery, characterized in that the dosage of the decusin and decosinol angelate is 0.5 mg / kg to 10 mg / kg.
4. The method of claim 3,

   The decosin and decosinol angelate is more preferably from 0.5 mg / kg to 2 mg / kg dose of leukocyte reduction and leukocyte recovery useful for the recovery of leukocytes.

Images