KR101380970B1 - Pharmaceutical composition for the treatment or the prevention of arthritis and leukemia comprising medicinal herb extract - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating arthritis, which comprises extracts of herbal medicine such as astringent, dried and salted, and a pharmaceutical composition for treating an anti-cancer agent.
The composition of the present invention can inhibit the production of TNF-? Which plays an important role in the pathogenesis of arthritis as a signal transduction substance, and thus can be widely used for the prevention and treatment of arthritis. Furthermore, it exhibits anti-blood-cancer activity against a human-derived blood cancer cell line, and can be widely used as an anticancer agent made of a natural substance.

Description

[0001] The present invention relates to a pharmaceutical composition for the prevention or treatment of arthritis and an anti-blood-derived cancer comprising an herbal extract and a pharmaceutical composition for treating or preventing arthritis and leukemia comprising medicinal herb extract,

The present invention relates to a pharmaceutical composition for preventing or treating arthritis, which comprises extracts of herbal medicine such as astringent, dried and salted, and a pharmaceutical composition for treating an anti-cancer agent.

Arthritis occurs in various forms, such as rheumatoid arthritis and joint inflammation-related diseases, especially chronic polyarthritis caused by inflammatory changes in the synovial membrane of the lining of the articular capsule can cause swelling and pain in the joints of the body, and in severe cases can become a physically disabled person. Is a chronic disease. Arthritic conditions, such as rheumatoid arthritis, are also progressive and can cause joint disorders such as deformity and joint indomitability, often resulting in severe physical disability due to a lack of effective treatment and continued deterioration.

The direct cause of arthritis is not clear yet, and researches on it have been continuing. To date, no effective preventive measures against arthritis such as rheumatoid arthritis have been known. There are a lot of things to do.

Traditionally, the treatment of arthritis includes steroids such as cortisone and other corticosteroids, non-steroidal anti-inflammatory agents such as aspirin, piroxycam and indomethacin, gold agonists such as Orotioctic acid, chloroquinone agents and D- Anti-rheumatic agents such as cilamine, anti-rheumatic agents such as silamine, gout inhibitors such as colchicine, and immunosuppressants such as cyclophosphamide, azathioprine, methotrexate and rebamisole.

However, the above-mentioned therapies are not a fundamental therapy, and steroid hormones have been widely used as a remedy for arthritis, but their use has become difficult as their side effects become clear.

In addition, arthritis, especially chronic rheumatoid arthritis, causes severe pain to the patient. Therefore, it is necessary to take anti-inflammatory drugs and so on. So far, aspirin agent and botazoline agent have been widely used for relieving such pain and swelling of joints Has been used. However, since aspirin, etc., have a fatal impact on humans, it is difficult to continue taking the amount needed to treat arthritis.

These existing chemotherapeutic agents have drawbacks such as side effects that interfere with long-term use of drugs, deficiency of anti-inflammatory effects, and lack of efficacy against arthritis that has already occurred, and currently, indomethacin Ginsenosides, furpenic acid and non-steroidal anti-inflammatory agents have been prepared and used.

Therefore, it is desired to develop a therapeutic agent for arthritis which shows a symptomatic effect on acute inflammatory symptom and pain and a solution to such a problem. Although most of the arthritic drugs currently used are different in degree or individual, And especially for the treatment of rheumatoid arthritis, it is very important to develop medicines with low side effects because they need to be taken for a long time.

In addition, some of the above medicines may be administered by intravenous injection or intraperitoneal injection. In this case, not only troublesome but repeated intravenous injection may cause not only allergy, shock but also hygiene problems. Therefore, there is a need for a safe and easy-to-eat therapeutic agent.

On the other hand, the development of anticancer drugs is the most important point in the development of biotechnology-related technology. Cancer is one of the causes of death in the United States and Japan as well as Korea, and accounts for more than half of the mortality rate in large hospitals. As part of efforts to conquer these cancers, many advances have been made in the diagnosis of early detection of cancer, and the development of surgery, radiation therapy, anticancer drugs, and biological methods have been developed in cancer therapy. Among them, many anti-cancer drugs have been tried with various kinds of cancer cells. However, until now, there has been no anti-cancer drug as a true therapeutic agent. However, only an adjuvant treatment or a drug for helping life extension for a short time has been developed.

The anti-cancer agents used mainly in the clinic are substances that exhibit cytotoxicity by reacting with nucleic acid synthesis inhibitors such as 5-fluorouracil and citrarabine or nucleic acids such as alkylated drugs, mitomycin, cisplatin, procarbazine, etc. Can be classified. However, since these anticancer drugs have various side effects, they have many problems, and there is an urgent need for the development of new cancer treatments that solve these problems.

Accordingly, the present inventors have studied the preventive or therapeutic effects of arthritis and blood cancer in natural medicinal herbs. As a result, the present inventors have found that a combination composition of mussel, alfalfa and salmon inhibits TNF-α production and inhibits anti- , Thereby completing the present invention.

Accordingly, an object of the present invention is to provide a combination composition containing astringent, astringent and bongline which can be used for prevention or treatment of arthritis by inhibiting the production of TNF-α.

It is another object of the present invention to provide a composition containing astringent, astringent and bongline which can be used for prevention or treatment of cancer by exhibiting antitumor activity against a blood cancer cell line.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for preventing or treating arthritis, which comprises astaxanthin, lysine and salicylic acid as an active ingredient.

The present invention also provides a pharmaceutical composition for anti-hematological malignancies comprising as an active ingredient astaxanthin, salicylate and salicylate.

The composition of the present invention can inhibit the production of TNF-? Which plays an important role in the pathogenesis of arthritis as a signal transduction substance, and thus can be widely used for the prevention and treatment of arthritis. In addition, the composition exhibits anti-blood-cancer activity against a human-derived blood cancer cell line, and can be widely used as an anticancer agent made of a natural substance.

Hereinafter, the present invention will be described in detail.

The present invention is characterized by a pharmaceutical composition for preventing or treating arthritis and a pharmaceutical composition for treating an anti-hematocarcinoma, the composition comprising rumen, alopecia and saliva as an active ingredient.

The Achyranthis Bidentatae Radix is a root of Achyranthes fauriei LEVEILLE et VANIOT or Achyranthes bidentata Blume, which is a plant of Amarantha ceae and a height of about 90 cm. The wax contains water-decomposable oleanolic acid and saponin decomposed into glucuronic acid, and alkaloids such as aralantine (C6H11O2N2H2O), which is soluble in water, and other alkaloids, and a large amount of mucus. The ash content of the water extract is about 8% and the potassium salt is 24.5%. In addition, it contains polybasic acids including amino acids such as sitosterol, stigmasterol, and aspartic acid and succinic acid. There are analgesic effects such as analgesic effect, antispasmodic effect, diuretic effect, antiallergic effect, etc. (Jang Chang Soo et al., Modern Medicine, Seoul, Korea, 24-128, 1993)

The medicines used in the treatment of bronchial asthma are medicines such as medicines for the treatment of schizophrenia, drugs for urinary incontinence, medicines for the use of medicines, and medicines for medicinal products such as medicines.

Achyranthes japonica (MIQ.) NAKAI and Achyranthes bidentata Blume can be used for the above-mentioned nutrients.

In addition, the component of the composition of the present invention refers to a rootstock or juniper of Atractylodes japanica Koidzumi , Compositae or Atractylodes ovata Koidzumi, Compositae . In oriental medicine, it is said to be used as a directional cardiopulmonary agent to reinforce the spleen, and it is known to have sedative, diuretic, gifted, nurturing, and anti-anesthetic effects. Also, it can be applied to Sajunjae Tang, Double Tang, Bojungwang Tang, Dangwi Mountain, Kwakyangjeongsan Mountain, Ohryeongsan, Myeongryeon Mountain, Yiwangtang, and Yangsuwangtang.

And another component of the composition of the present invention, balsam ( Impatiens balsamina ), also known as balsam, is native to India and Southeast Asia, grows well in sunlight and grows relatively well in bad environments. It grows well in wetlands, so it is planted in wet clay and does not dry in summer. There are high grade seeds which are more than 60cm in height and dwarf seeds which are as low as 25 ~ 40cm. They grow straight and are fleshy and the nodes of the bottom are especially prominent. Leaves are alternate phyllotaxis, with petiole, narrow on both ends, with sawtooth on edge. If you sow in April ~ May, flowers will start to flower after June. There are 2 ~ 3 flowers on each axilla, and there is a peduncle. It is sagged downward. The petals spread wide to the left and the right. There are pink, red, vermilion, purple, and white, and the flower shape is single flower, double flower. There are 5 stamens, anther is connected to each other, and hair is in the ovary.

In the present invention, the bongline may be a whole plant, and a part such as roots, stems, leaves or flowers may be used, and preferably leaves, stems, and flowers are preferably used.

The composition of the present invention is preferably composed of extracts obtained by drying, finishing, and then extracting with water, a C ₁ -C ₄ alcohol or a mixed solvent thereof, but it is preferred that each of the extracts, Single extracts may be mixed.

The extract according to the present invention can be produced by a conventional extraction method such as supercritical extraction, subcritical extraction, high-temperature extraction, high-pressure extraction, ultrasonic extraction or other adsorption methods using XAD and HP-20 May be used. Preferably, the mixture may be heated and refluxed or extracted at room temperature. However, the present invention is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 times, but is not limited thereto.

For example, the complex extract according to the present invention is prepared by pulverizing the dried, dried and boiled starch to an appropriate size, placing it in an extraction container, adding an extraction solvent, boiling the solution, refluxing the extract, allowing it to stand for a predetermined time, Can be obtained. The extraction time is preferably 1 to 20 hours, more preferably 5 to 15 hours. Thereafter, a method such as concentration or lyophilization may be additionally performed.

At this time, the blending ratio of the wax, the dried wax and the boiled wax is not particularly limited, but preferably 20 to 500 parts by weight, more preferably 50 to 200 parts by weight, respectively, based on 100 parts by weight of the wax .

Meanwhile, the composition of the present invention may further include at least one selected from the group consisting of sea tangle, hemp blood, etc., and the content thereof is preferably 30 to 200 parts by weight based on 100 parts by weight of the sea tangle .

The composition of the present invention can be prepared by fermenting the raw material by natural fermentation or by treating a fermenting microorganism such as malt, yeast, grape, wine fermentation enzyme source or yeast. In particular, fermentation of the composition of the present invention can have a positive effect on the maintenance of the efficacy, taste and form of the final formulation. In addition, the fermented rice with malt or malt, and then filtered to take a supernatant may be fermented into the extract.

When using malt, a large amount of starch already contained by the malt enzyme can be saccharified to obtain an extract suitable for fermentation, and as a supplement, malt starch supplements are used up to 50% of malt. In this case, the use of inexpensive supplementary materials can be expected to reduce the fermentation time and increase the fermentation efficiency economically such as increase in the fermentation extraction amount, as well as a side effect such as improving the flavor and reducing turbidity. Examples of the additive that can be used include refined starch such as corn, rice, wheat, papain (obtained from the liquid of Carica papaya), kiwi or pear, refined starch, There is also a glucose syrup like corn syrup.

Nuruk (grain) is wheat and rice flour is usually used as a raw material, green beans, glutinous rice, barley, etc. are also used.

Yeast sources include air, dokmari, barley straw, straw, mulberry, wormwood, resident, lotus, young leaves of pine needles and pine needles. There are roasted, steamed, lightly steamed, and wheatgrass, and the grain size of the raw material can be used from 12% to differential. Types of additives include mulberry leaves, wormwood, lactose juice, peach seed powder, apricot seed powder and melon powder, which are prepared by treating with a coenzyme. The coenzymatic agent is the one which cultivated bacterium such as Rhyzopus, Usami and 0ryzae which are the bacterium of the native leukocyte and increased the saccharification power, and pursues the complex taste and the saccharification power of natural leu.

In the present invention, the pharmaceutical composition may be prepared by various methods.

The most typical manufacturing process in the present invention will be described as an embodiment.

For example,

(1) 140-180 parts by weight of wax, 140-180 parts by weight of boiling water, 140-180 parts by weight of boiling water and 80-120 parts by weight of malt are used, boiling, boiling, and boiling water are heated to boiling for 2-5 hours do. Adding an enzyme solution to about 15-30% by weight, heating for 6-10 hours, and then filtering and taking a filtrate;

(2) adding 40 to 60% alcohol to the residue obtained in the above step (1), allowing to stand for 10-15 hours, filtering, removing the residue and heating the filtrate to obtain an alcohol solution;

(3) adding about 15 to 30% by weight of the enzyme solution to the solution obtained in the step (2), fermenting the solution for 6 to 12 hours, and then filtering to obtain a filtrate; And

(4) The filtrate obtained in the above step (1) and the filtrate obtained in the above step (3) are combined in a ratio of 40 to 60 wt% and 60 to 40 wt%, respectively, and 20 to 30 wt% Fermenting for 8 to 24 hours and concentrating the filtrate;

The object of the present invention can be produced.

Over-production of TNF-α (Tumor necrosis factor-alpha) has been implicated in progression of inflammation, cell differentiation, and destruction of joints, leading to the onset of arthritis (Douni E. J. Inflamm. 1995, 47, 27). The composition of the present invention is effective for preventing and treating arthritis because it can inhibit the production of TNF-a.

In addition, the composition of the present invention has been shown to exhibit strong activity against cancer cell lines, and exhibits excellent anticancer activity against blood cancer cell lines, and thus can be used as an anticancer drug.

In addition, the present invention was observed to have an anti-inflammatory, anti-viral and immuno-enhancing effect through the above effects.

Accordingly, the present invention includes a therapeutic agent for arthritis, an anti-blood cancer agent, an antiviral agent, an anti-inflammatory agent, and an immunity enhancer, which comprises the above pharmaceutical composition as an active ingredient.

The pharmaceutical composition of the present invention may be formulated into various oral or parenteral dosage forms described below, but the present invention is not limited thereto.

Formulations for oral administration include, for example, tablets, pills, hard / soft capsules. Liquid. Suspensions, emulsifiers, syrups. Granules, elixirs, and the like, these formulations may be used one or more diluents or excipients, such as fillers, extenders, wetting agents, disintegrants, lubricants, binders, surfactants, etc. that are commonly used in addition to the active ingredient. As a disintegrant, agar, starch, alginic acid or its sodium salt, calcium monohydrogen phosphate anhydride, etc. may be used, and as lubricant, silica, talc, stearic acid or its magnesium salt or calcium salt, polyethylene glycol, etc. may be used. As a binder, magnesium aluminum silicate. Starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidine, low-substituted hydroxypropylcellulose, and the like can be used. In addition to lactose, dextrose, sucrose, mannitol, sorbitol, cellulose. Glycine or the like may be used as a diluent, and in some cases, generally known boiling mixtures, absorbents, colorants, flavors, sweeteners, and the like may be used together.

In addition, the pharmaceutical composition may be administered parenterally, and parenteral administration may be performed by injecting subcutaneous injection, intravenous injection, intramuscular injection, or intra-thoracic injection. In this case, for formulation into parenteral administration form, the extract or its fraction may be mixed with water or a stabilizer or buffer to prepare a solution or suspension, which may be prepared into a unit dosage form of an ampule or vial.

The composition may be sterile or contain preservatives, stabilizers, hydrating or emulsifiers, salts for controlling osmotic pressure, buffers and other auxiliaries and other therapeutically useful materials, which are conventional methods of mixing, granulating or coating methods. It can be formulated accordingly.

When the pharmaceutical composition of the present invention is formulated in a unit dosage form, it is preferable that the mixed extract of raspberry, vermiculite and persimmon as an active ingredient contains a unit dose of about 0.1 to 1,500 mg. The dosage depends on the physician's prescription depending on factors such as the patient's weight, age and the particular nature and severity of the disease. However, the dosage required for adult treatment usually ranges from about 1 to 500 mg per day, depending on the frequency and intensity of administration. For intramuscular or intravenous administration to adults, a single dose separate from the usual dosage of usually about 5 to 300 mg per day would be sufficient, but for some patients a higher daily dose may be desirable.

Hereinafter, the present invention will be described in detail with reference to Examples and Production Examples.

EXAMPLES The following examples and preparations are merely illustrative of the present invention, and the contents of the present invention are not limited by the following examples and preparative examples.

Example 1: Preparation of a complex extract

160 g of washed and dried shrubs, 160 g of balsam and 160 g of balsam were cut into a 5 L round-bottomed flask, and 2 L of purified water was added thereto. The resulting mixture was refluxed with a reflux condenser and extracted with a 80-100 ° C water bath for 10 hours to obtain an extract.

Example 2: Preparation of a complex extract

160 g of washed and dried shrubs, 160 g of balsam and 160 g of balsam were cut into a 5 L round-bottomed flask, and 2 L of purified water was added thereto. The resulting mixture was refluxed with a reflux condenser and extracted with a 80-100 ° C water bath for 10 hours to obtain an extract. The extract was cooled to about 50 ° C., and then 12 g of malt and 32 g of rice were fermented at room temperature for 12 hours. The fermented extract was heated to 100 DEG C and filtered at high temperature, and the solvent was evaporated and concentrated to obtain a final extract.

Example 3: Preparation of a complex extract

An extract was obtained in the same manner as in Preparation Example 2, except that 160 g of dried sea tangle (160 g) was further added to 160 g of dried persimmon, 160 g of dried seaweed and 160 g of salted seaweed.

Example 4: Preparation of complex extract

(1) First, water, liquor, and boiling water are added to water in the above composition, heated and boiled for 3 hours and cooled. The enzyme solution was added thereto to about 20% by weight and heated for 8 hours, followed by filtration to obtain a filtrate.

(2) 40-60% alcohol is added to the residue obtained in the above step (1) and left for 10-15 hours. Then, the residue is discarded and the filtrate is heated to blow off the alcohol to obtain a solution.

(3) About 20% by weight of the enzyme solution is added to the solution obtained in the step (2) and fermented for 8 hours and then filtered to obtain a filtrate.

(4) The filtrate obtained in the above step (1) and the filtrate obtained in the above step (3) were combined at a ratio of 50:50, and 25% by weight of the malt was mixed and then fermented for 12 hours and the filtrate was concentrated The extract is prepared.

Experimental Example 1: Measurement of inhibitory activity of LPS-induced TNF-α production

Six weeks old C57BL / 6 mice (average body weight 15.0-19.0 g, total 20) supplied from Orient Co. were purified for one week. The mice were divided into 5 groups and the extract solution of the above example was administered at 10 mg / kg twice in 24 hours at intervals of 4 hours. After 2 hours from the administration, 400 ml / L of lipopolysaccharide (lipid polysaccharide) was intraperitoneally administered to four groups of mice to which the extract of the above example was administered, in an amount of 0.1 ml per mouse. 90 minutes after the administration of the LPS, blood was collected from the orbital vein and centrifuged (6000 rpm, 5 minutes) to separate the plasma.

The separated plasma was dispensed into an immune-plate coated with anti-TNF-α antibody to bind TNF-α in the plasma to the antibody, and color development was induced using a TMB substrate solution Then, the color development was stopped with stop solution (1M HSO). The concentration of TNF-α was then determined by subtracting the absorbance at 540 nm from 450 nm with ELISA-reader. The results are shown in Table 1 below.

group TNF-a (pg / ml) Suppression efficiency (%) LPS non-inferiority group -107.6 ± 94.3 LPS induction No treatment 4885.3 ± 371.5 Example 1 Administration 2291.7 ± 175.4 53.1 Example 2 Administration 2352.2 ± 377.4 51.9 Example 3 Administration 2403.8 ± 326.3 50.8

As shown in Table 1, it was confirmed that the extract of the Example can inhibit TNF-α production by 50% or more in LPS-induced mice. Therefore, it was confirmed that the composition of the present invention inhibits the generation of TNF-α, which is an inflammation-inducing factor, to prevent or treat the development of arthritis or to have anti-inflammatory and antiviral therapeutic effects.

Experimental Example 2: Measurement of anti-blood cancer activity

Human-derived blood cancer cell line HL60, which was frozen in liquid nitrogen, was thawed and cultured in a CO ₂ incubator at 37 ° C and 5% CO ₂ concentration.

The cultured cell line was adjusted to a cell concentration of 4 × 10 ⁷ cells / ml with PBS (Phosphate Buffered Saline) to prepare a cell culture solution.

On the other hand, five-week-old SPF nude mice (average body weight 16.0 to 19.0 g, total 30) supplied from Orient Co., Ltd. were purified for one week. The mice were divided into three groups, and 0.3 ml of the cell culture solution was injected into the mice of all the groups under the subcapsular region between the right clavicular and chest wall.

Then, when the volume of the average amount of each group reached about 50.0 mm ³ , mice were fixed by the subcutaneous skin fixation method, and the extract of Example 4 was orally administered to one group of mice at a dose of 250 mg / kg every day, As a positive control group, adriamycin was administered to mice of other groups at a dose of 2 mg / kg every day.

The size of the tumor was determined for 4 days after the administration of the drug. The size of the tumor was measured by measuring the length, height, and width in three directions using a veneer caliper.

[Equation 1]

Tumor size = (length × width × height) / 2

Tumor measurement results are shown in Table 2 below.

group Tumor size (mm ³ ) 0 days 2 days 4 days No treatment 0 67.5 ± 65.5 175.8 ± 122.2 Example 4 Administration 0 44.9 ± 16.3 157.6 ± 59.4 ADR administration 0 35.1 ± 14.3 147.3 ± 59.4

As shown in Table 2, the tumor growth inhibitory effect was about 33% similar to that of the positive control in the early stage (2 days) of tumor growth. The composition of the present invention showed anti-blood cancer activity, It was confirmed that there was an effect due to On the other hand, no abnormal symptoms were observed upon administration of the composition of Example 4 in the above experiment.

Claims (14)

delete delete delete delete delete delete delete Pharmaceutical composition for an anti-blood cancer drug, characterized in that it comprises an extract of the fermentation products of the fermentation products of dew, baekchul and Bongseon as malt.
The method of claim 8 wherein wooseul, baekchul bongseon and water, wherein comprising the extract extracted with an alcohol or a mixed solvent of a C ₁ ~ C ₄ jeyong blood cancer pharmaceutical composition.
The pharmaceutical composition of claim 8, wherein the baekchul and bongseon each include 20 to 500 parts by weight based on 100 parts by weight of wedge.
delete The pharmaceutical composition of claim 8, wherein the fermentation is performed using one or two or more fermentation sources selected from yeast, grape and wine fermentation enzyme sources and yeast.
The pharmaceutical composition for anti-blood cancer drug according to claim 8, further comprising thawed blood.
(1) 140-180 parts by weight of wax, 140-180 parts by weight of boiling water, 140-180 parts by weight of boiling water and 80-120 parts by weight of malt are used, boiling, boiling and boiling are heated in boiling for 2-5 hours Adding the enzyme solution so that the enzyme solution is 15 to 30 wt%, heating the solution for 6 to 10 hours, and then filtering to take the filtrate;
(2) adding 40 to 60% alcohol to the residue obtained in the above step (1), allowing to stand for 10-15 hours, filtering and heating the filtrate to remove alcohol to obtain a solution;
(3) adding 15 to 30% by weight of an enzyme solution to the solution obtained in the step (2), fermenting the solution for 6 to 12 hours, and then filtering to obtain a filtrate; And
(4) The filtrate obtained in the above step (1) and the filtrate obtained in the above step (3) are combined in a ratio of 40 to 60% by weight and 60 to 40% by weight, respectively, and 20 to 30% Followed by fermentation for 8 to 24 hours and concentration of the filtrate;
Method for producing a pharmaceutical composition for anti-blood cancer comprising a.