KR20100128508A - Dual core tablets with pharmaceutical composition for preventing and treating disease related to renal failure - Google Patents

Abstract

PURPOSE: A dual core tablet for preventing and treating renal insufficiency is provided to enhance and protect gastrointestinal movement and to maximize treatment effects. CONSTITUTION: A dual core tablet for preventing and treating renal insufficiency contains an enteric core tablet and gastric external tablet. An enteric coat is applied between the enteric core table and gastric external tablet. The enteric core tablet contains herb extract of Salviae miltiorrhizae radix, Pinelliae rhizome, Coptis rhizome, Evodiae fructus, Epimedii herba, Rhei radix et rhizome, and Polyporus. The gastric external tablet contains herb mixture extract of Codonopsis pilosulae radix, Perillae folium, Glycyrrhizae radix, Artemisiae capillaris herba, Alismatis rhizome, Poria, Atractylodis macrocephalae rhizome, and Cinnamomi ramulus.

Description

Dual core tablets with pharmaceutical composition for preventing and treating disease related to renal failure}

The present invention relates to a double-nucleated tablet for the prevention and treatment of renal failure, and more specifically, enteric core tablets comprising a mixed extract of Chinese herbal medicine consisting of Salvia, Banja, Hwangyeon, Osuyu, Eumyangkwa, Rhubarb, and herb Gastric protective and strengthening ingredients surrounded by gastric external tablets containing a mixed extract of Chinese medicine consisting of ginseng, lobular, licorice, injin, taxach, baekbokyeong, Baekchul and Gyeji, tablets, enteric between the enteric core tablets and stomach tablets It relates to a double-nucleated tablet for the prevention and treatment of renal failure characterized in that the blood is coated.

In general, the chief function of the kidneys is to filter waste and make urine while balancing the body's water, electrolytes, acids and bases to keep body fluids in proper condition, and calcium and phosphorus to keep bones strong. And, hematopoietic hormones to prevent anemia, while secreting hormones that control blood pressure to maintain normal blood pressure.

However, for a variety of reasons, the function of these kidneys can be reduced. In case of chronic kidney failure, the excretory function of the kidneys is reduced to 25% or less of normal. Nausea, anorexia, nocturnal enuresis that occurs early in the uremia It is difficult to prevent or diagnose at first because it is easy to pass, such as sleep disorders, fatigue, indigestion, etc., and if kidney failure develops severely, you may smell urine when you breathe, swelling, itching, memory loss, etc. Symptoms such as difficulty, convulsions, and lethargy occur.

Although chronic renal failure is so severe, there are currently no suitable treatments for treating such renal failure (Wesołowski P, Saracyn M, Nowak Z, Wakokocz Z, Peritoneal dialysis and insulin resistance in patients with chronic kidney disease due to nondiabetic nephropathy , Pol.Merkur.Lekarski. , 24, 359-363, 2008).

Chronic renal failure is mainly due to inflammatory reactions due to fibrosis of kidney cells and excessive increase of macrophages, and it is known that inhibiting inflammation can prevent renal cell destruction by reducing fibrosis of kidney cells (Naiki). Y, Maeda Y, Matsuo K, Yonekawa S, Sakaguchi M, Iwamoto I, Hasegawa H, Kanamaru A, Involvement of TGF-beta signal for peritoneal sclerosing in continuous ambulatory peritoneal dialysis, J. Nephrol. , 16, 95-102, 2003 Therefore, the treatment of chronic renal failure is currently using a therapeutic agent that can exhibit an anti-inflammatory effect. When the anti-inflammatory effect is used as a treatment for chronic renal failure, there is a disadvantage of continuously taking anti-inflammatory drugs, as well as side effects.

Therefore, the present inventors have made a diligent research to treat chronic renal failure through research on herbal medicines that can inhibit the production of inflammatory mediators secreted to cause inflammation during macrophage inflammatory reactions. It was possible to invent a pharmaceutical composition for the treatment of herbal renal failure, which comprises extracts extracted from Chinese medicine, which are mixed with Hwangyeon, Osuyu, Yin-Yang, Rhubarb, Lobules, Licorice, Injin, Taxa, Paekbokyeong, Baekchul, Seolji and Gyeji at a specific ratio. .

The pharmaceutical compositions of the herbal treatment for chronic renal failure may be eluted and absorbed at the same time, but especially in patients with renal failure who have weak gastrointestinal function, activation of gastrointestinal function needs to be performed first.

Therefore, the present inventors first look at the functions of the respective herbal medicines constituting the pharmaceutical composition for the treatment of oriental renal failure, and among them, selected herbal medicines effective in the activation of gastrointestinal function. The corresponding functions of herbal medicines are as follows.

Ginseng (Codonopsis pilosulae radix, scientific name: Codonopsis pilosula FR. NANNF) is used as a medicinal herb by drying its roots. It is non-toxic and keeps spleen, produces essences, and grows brown. It acts as a kind of tonic to stop, and is known to be used for symptoms such as weak stomach weakness, loss of appetite, limb disability, mental anxiety, fatigue, ruined seawater, and smut.

Lobules (pharmaceutical name: Perillae folium, scientific name: Perilla frutescens L. BRITT.) Are known for their effects on blood circulation, elimination of inflammation, and digestion.

Licorice (medicinal name: Glycyrrhizae radix, scientific name: Glycyrrhiza uralensis FISCH) is a licorice commonly used as a medicinal herb. In Dongbogam, it controls the heat and fraud of the five viscera and the eyes, nose, mouth, ears and faeces. It is known to normalize physiological phenomena, communicate all blood vessels, strengthen muscles and bones, improve nutrition, detoxify the toxicity of all medicines, and show the efficacy of the combined herbal medicines.

Injin (Pharmaceutical name: Artemisiae capillaris herba, scientific name: Artemisia capillaris THUNB) is used to treat fever in Chinese medicine, fever, moonshine, faeces, and urine The efficacy of is known.

Taxa (medicinal name: Alismatis rhizome, scientific name: Alisma plantago-aquatica var. Orientale SAMUELS) is non-toxic and mainly acts on the kidneys and bladder to release urine, release moisture and reduce heat.

Baek Bok-Ryung (People's name: Poria, scientific name: Poria cocos SCHW) is known to be used for the symptoms of swollen or indigestion or difficulty urinating due to weak spleen or stomach.

The medicinal herb (Atractylodis macrocephalae rhizome, scientific name: Atractylodes macrocephala KOIDZ) is an aromatic medicinal agent used in indigestion.It is used for indigestion or to drain excess water from the body. It is known to be used to treat symptoms.

Gyeji (Chemical name: Cinnamomi ramulus, scientific name: Cinnamomum cassia PRESL) is an aromatic sensation agent, which is used as an appetite enhancer by combining the powder with other powders, or in the Chinese medicine, including a cold. It is used for symptoms such as circulatory diseases and acute fever.

As mentioned above, ginseng, lobule, licorice, injin, taxa, baekbokyeong, Baekchul, gyeji etc. can be used to improve indigestion and gastrointestinal function as a dry placebo to relax and warm the stomach, thereby protecting and strengthening gastrointestinal movement Since it induces elution and absorption of sweet ginseng, hwangja, rhubarb, sorghum, eumyanggwa, rhubarb, and ghosts, it was necessary to elute separately from stomach and intestine to exert these effects.

Thus, the present inventors, so that the ginseng, lobules, licorice, licorice, injin, tack, baekbokyeong, baekryeok, gyegeok and the like in the pharmaceutical composition of the herbal treatment for chronic renal failure in the first place, and then eluted and absorbed in the stomach, dansam, Banja, Hwangyeon, Ohsuyu In the case of using a double-nucleated tablet formulation, so that yin and yang, rhubarb, and lysate can be eluted and absorbed in the duodenum and small intestine, it was confirmed that the above object can be achieved and completed the present invention.

Therefore, the main object of the present invention is a pharmaceutical composition for preventing and treating renal failure, consisting of extracts of ginseng, red ginseng, halved, yellow lotus, sesame oil, yin and yang, rhubarb, lobule, licorice, jinjin, taksa, baekbokyeong, baekrye, yeongji, gyeji The ginseng, leaflet, licorice, injin, taksa, baekbokyeong, baekchul, gyeji components that have the effect of protecting and strengthening the gastrointestinal motility, the remaining components of the sweet ginseng, halve, rhubarb, sessile, yin and yang, rhubarb, and herb are duodenum and small intestine. By eluting in to maximize the prevention and treatment of renal failure.

According to an aspect of the present invention, the present invention is enteric core tablet comprising a mixed extract of herbal medicine consisting of Salvia, Banja, Hwangyeon, Ohsyuyu, Eumyangkwa, Rhubarb and Sesame as an ingredient for preventing and treating renal failure and gastrointestinal protection surrounded by Gastric external tablets containing a mixed extract of Chinese herbal medicine consisting of ginseng, lobule, licorice, injin, taxa, baekbokyeong, Baekchul and Gyeji are compressed as a reinforcing ingredient, and enteric coating is coated between the enteric core tablet and the stomach external tablet. It provides a dual core tablet for the prevention and treatment of renal failure characterized in.

In the double-nucleated tablet for preventing and treating renal failure of the present invention, when the enteric core tablet is 100% of the total length of the enteric core tablet and the gastric external tablet, 6 to 10% by weight of salviae, 4 to 8% by weight, and 3 to 5 kg of yellow lotus It is preferable to include a mixed extract of the Chinese herbal medicine consisting of 5% by weight, sewage oil 3-5% by weight, 8-12% by weight of Eumyangkwa, 4-8% by weight of rhubarb and 4-6% by weight.

In the double-nucleated tablet for preventing and treating renal failure of the present invention, when the total external tablets for the stomach are 100% of the total length of the enteric nuclear tablets and the stomach external tablets, 6 to 10% by weight of ginseng, 4 to 8% by weight of lobules, and licorice 2 It is preferred to include a mixed extract of the herbal medicine consisting of 4 to 4% by weight, phosphorus 15 to 20% by weight, 10 to 14% by weight of tack, 4 to 6% by weight of Baekbokyeong, 4 to 6% by weight of Baekchul and 2-3% by weight of ginseng. Do.

In the present invention, the extract is preferably prepared by freeze-drying the hot water extract of the herbal medicine.

Hereinafter, the dual nuclear tablet for preventing and treating renal failure of the present invention will be described in more detail step by step.

The present invention protects and enhances gastrointestinal motility among extracts of 15 kinds of mixed medicinal herbs consisting of Mansam, Dansam, Banja, Hwangyeon, Osuyu, Yinyanggok, Rhubarb, Lobules, Licorice, Injin, Taxa, Paekbokyeong, Baekchul, Haeyoung and Gyeji. Ginseng, lobules, licorice, Injin, Taeksa, Baekbokryeong, Baekchul, Gyeji extract as stomach external tablets, and the rest of Dansam, Banja, Hwangyeon, Osuyu, Yin Yang Guk, rhubarb, sesame extract as a double core tablet When prepared by taking ginseng, lobule, licorice, injin, taxa, baekbokyeong, baekchul, gyecheol components in the stomach to protect the stomach and enhance the function and in the duodenum and small intestine, salvia, hwangja, rye lotus, oosuyu, yin and yang, Rhubarb, aging component is eluted to the dual nucleus tablet for the prevention and treatment of renal failure that can enhance the prevention and treatment of renal failure.

In particular, the herbal extract of the present invention is extracted from natural products, even if taken for a long time, the side effects are insignificant, TGF-β (tumor growth factor-β), which induces fibrosis of inflammatory cytokines and tissues that cause inflammation, Treatment of renal failure by the antioxidant effect of inhibiting the expression of Smad2 (Sma and Mad-related protein2), α-Smooth muscle actin (α-SMA) and proliferative cell nuclear antigen (PCNA) Prevent.

At this time, the water used to extract the mixed herbal medicine in the present invention refers to water suitable for drinking standards based on the Ministry of Health, generally water or tap water, such as purified water, sterile water, anion water filtered through a filter or water purifier, etc. , Bottled water and the like can be used. However, the solvent used to extract the active ingredient of the mixed herbal medicine in the present invention is not limited to water, and if using a device that removes to the extent that it is not harmful to the human body, organic solvents such as methanol, ethanol, acetone, diethyl ether, etc. Can be used.

In addition, ginseng (Codonopsis pilosulae radix, scientific name: Codonopsis pilosula FR. NANNF) is used as a herbal medicine by drying the roots. It is known to be used as a kind of tonic to stop the symptoms such as weak stomach weakness, loss of appetite, limb force, mental anxiety, fatigue, ruined seawater, and smut. In the present invention plays a role in addition to the above efficacy. At this time, in the present invention, it is usually preferable to use one that has been regulated with wheat flour.

Salviae (salviae miltiorrhizae radix, scientific name: Salvia miltiorrhiza BGE) is used as a herbal medicine by drying its roots.It is a tonic, painkiller, analgesic, anti-inflammatory, and hemostatic drug in Chinese medicine, dysmenorrhea, abdominal pain, menopause, postpartum pain, It is known to be used for rheumatic symptoms. In the present invention plays a role in addition to the above efficacy. At this time, in the present invention, it is preferable to use a thing that legally regulated sweet ginseng.

Banha (pharmaceutical name: Pinelliae rhizome, scientific name: Pinellia ternata THUNB. In general, it is preferable to use a half-drug that is regulated with ginger to remove toxicity. In the present invention, it serves as an expectorant together with Baekbokryeong.

It is known that Hunan (Coral, Coptis rhizome, scientific name: Coptis chinensis FRANCH) is used as an anti-inflammatory agent or sedative for nervous anxiety, hyperemia, and inflammation. In the present invention serves to suppress the occurrence of inflammation derived from the renal failure.

Oh Su-Yu (Evodiae fructus, scientific name: Evodia rutaecarpa (Juss.) Benth.] Is a medicine that comforts and warms the stomach in agreement. It is known that, in the present invention, it plays a role in activating the function of the kidney with other herbal medicines.

Eum Yang-Kwak (Epimedii herba, scientific name: Epimedium koreanum NAKAI) has jeongjeong, anti-inflammatory, analgesic, diuretic, strengthens the kidneys, strengthens the kidneys, bones, strength, sexual function (Hyangjeombangbang), It is known to have an improvement effect (agreement) on infertility, cold sickness, wind sickness, weakness and forgetfulness. In the present invention serves to protect the body weakened by kidney failure.

Rhubarb (pharmaceutical name: Rhei radix et rhizoma, scientific name: Rheum palmatum L.) is used as an indigestion, chronic gastroenteritis, flatulence, ataxia and diarrhea as a medicine in the Chinese medicine, and it is used for the symptoms of constipation and habitual constipation. In addition to acts as an antipyretic. Usually, it is preferable to use the thing which burned and burned in ash.

Lobules (pharmaceutical name: Perillae folium, scientific name: Perilla frutescens L. BRITT.) Is known to smooth blood circulation, eliminate inflammation and digestion well, in the present invention is a substance that causes inflammation in the kidney along with rhubarb, rhubarb, etc. It serves to suppress the activity of.

Licorice (medicinal name: Glycyrrhizae radix, scientific name: Glycyrrhiza uralensis FISCH) is a licorice commonly used as a medicinal herb. In Dongbogam, it controls the heat and fraud of the five viscera and the eyes, nose, mouth, ears and faeces. It is known to normalize physiological phenomena, communicate all blood vessels, strengthen muscles and bones, improve nutrition, detoxify the toxicity of all medicines, and show the efficacy of the combined herbal medicines. In the present invention, the effects of rhubarb, lobules, phosphorus and the like appear well, and play a role of not appearing significant side effects even if continuously taken.

Injin (Pharmaceutical name: Artemisiae capillaris herba, scientific name: Artemisia capillaris THUNB.) Can be used as an antipyretic, herbal repellent, faeces, and urine sulphate in Chinese medicine. Efficacy is known, such as rhubarb in the present invention serves to facilitate the antipyretic and excretion.

Taxa (pharmaceutical name: Alismatis rhizome, scientific name: Alisma plantago-aquatica var. Orientale SAMUELS) is non-toxic and acts mainly on the kidneys and bladder, which is used to excrete urine, release moisture, and reduce heat. It plays a role of making it smooth. At this time, in this invention, it is preferable to use what was legally regulated with vinegar.

Baek Bok-Ryung (People's name: Poria, scientific name: Poria cocos SCHW.) Is known to be used for the symptoms of swelling or indigestion or difficulty in urination due to weak spleen or stomach in oriental medicine. Along with age, it provides a synergistic effect on urination and facilitates kidney function. Baekbokyeong in particular, with the half provides the effect of expectoration (祛痰).

Acupuncture (Atractylodis macrocephalae rhizome, scientific name: Atractylodes macrocephala KOIDZ.) Is an aromatic sensitizing agent used in Korean medicine for indigestion or to drain excess water from the body. It is known to be used to treat the symptoms, in the present invention serves to facilitate the renal function with the above mentioned Jinjin, Taeksaek, Baekbongryeong and low age.

Low age (medicinal name: Polyporus, scientific name: Polyporus umbellatus PERS. FRIES) is known to be mainly used for urinary disorders of the urinary system such as diuresis, antibacterial, anti-tumor, antipyretic, acute urethritis, in the present invention together with other herbal medicines The kidneys function to function properly.

Gyeji (cod name: Cinnamomi ramulus, scientific name: Cinnamomum cassia PRESL) is a fragrant dry stomach agent, which is used as an appetite enhancer by combining the powder with other powders. It is used for symptoms such as circulatory disease, acute fever, etc. In the present invention, in particular, it serves to make it inconvenient to eat with other herbal medicines.

However, the ingredients used in the present invention are not limited to the above-stated medicinal herbs, and may be used in other ways as long as the side effects are minimized as long as they meet the object of the present invention.

In the present invention, when the total amount of the enteric core tablets and the stomach external tablets is 100% as the herbal medicine extract of the enteric core tablets of the double core tablets, 6 to 10% by weight of salviae, 4 to 8% by weight, and 3 to 5 weight of yellow lotus %, Sewage oil 3 to 5% by weight, 8 to 12% by weight of Eumyangkwa, 4 to 8% by weight of rhubarb and 4 to 6% by weight, and 6 to 10% by weight of ginseng as a herbal extract of stomach antler, 4 to 8 leaflets By weight, licorice 2-4% by weight, phosphorus 15-20% by weight, 10-14% by weight of taxi, 4-6% by weight of Baekbokryeong, 4-6% by weight of Baekchul and 2-3% by weight, respectively, If it is used in excess of the above-mentioned maximum amount per herbal medicine, the other necessary ingredients cannot be used sufficiently, and if it is used below the minimum amount, the effective effect of each herbal medicine is reduced, and the above-mentioned range of use for each herbal medicine has the maximum effect on kidney failure. To me This is because naeneunde preferred. At this time, more specifically, enteric core tablets of Chinese herbal medicine extracts such as salsa 150g, half 100g, rhubarb 60g, sewage oil 60g, Yinyangkok 160g, rhubarb 100g, 80g of mixed herbal medicine and stomach medicinal herb extracts ginseng 150g, lobular 100g, It is advisable to use a mixture of herbal medicine consisting of 50 g of licorice, 300 g of phosphorus, 200 g of talc, 80 g of Baekbokryeong, 80 g of Baekchul, and 40 g of ginseng and hot water extracted by adding water.

However, in the present invention, in order to prevent or treat renal failure in addition to water, while extracting the active ingredient of the herbal medicine sufficiently harmless to the human body may be extracted using an organic solvent, and then subjected to a process of removing the organic solvent.

The double core tablet of the present invention is preferably prepared by coating the enteric shell between the enteric core tablet and the stomach external tablet so that the components inside the enteric core tablet are not eluted from the stomach and the small intestine. You can use any coating that is harmless to the human body that can withstand the strong acid in the gastric juice and then decompose under conditions of the duodenum and small intestine and elute the components of the enteric core tablet, but either Eudragit L100 or Eudragit S100 can be used. It is preferable to select or use Eudragit L100 and Eudragit S100 together.

In addition, excipients that are acceptable to the formulation criteria, such as microcrystalline cellulose, colloidal silicon dioxide, corn starch and disintegrants such as corn starch, crospovidone, pregelatinized starch and glidants such as glyceryl It may be formulated with behenate and epidermis such as Opadry and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example 1 Preparation of Extract for Preventing and Treating Renal Failure

end. material

Ginseng 150g, Salvia 150g, Half 100g, Rhubarb 60g, Sorghum 60g, Yinyangkok 160g, Rhubarb 100g, Small leaf 100g, Licorice 50g, Injin 300g, Taxa 200g, Baekbokyeong 80g, Baek 80g, Seon 80g, Gyeji 40g

I. Extract manufacturer

In about 1,710 g of the mixed herbal medicine prepared above, water was added about twice the volume of the herbal medicine, heated for 5 hours to extract hot water, and the extract obtained by filtering the extract with gauze was lyophilized to treat the chronic renal failure of the present invention. The composition was prepared. At this time, the yield was 18.5%.

The composition was named WHW extract and used as a medicament for effect verification while storing it at 4 ° C.

Example 2. Clinical Trial Effects

Eight volunteers (all men) of renal failure patients were given a composition of 171 g per day for three months (one person / 171 g / day / 3 months).

Then, volunteer blood was collected and measured for blood creatine and BUN (Blood Urea Nitrogen) before and after taking.

As a result, as shown in FIG. 1, BUN is not shown), and in chronic renal failure patients, creatine (Creatine) is 1.4 or more, and BUN is expressed in 24 or more. When the WHW extract of the present invention was taken, it was found to fall to normal.

Example 3. Effect through Cell Line and Animal Experiments

end. Anti-inflammatory effect

① Inhibitory effect of active nitrogen species

Nitric oxide (NO) is a free radical produced from macrophages activated after pathogen infection, and the production of an appropriate amount of active nitrogen species maintains vascular homeostasis and shows anticancer effects, but excessive amounts of active nitrogen species Rather, it is known to accelerate the development of acute and chronic inflammatory diseases by mediating the inflammatory response due to its strong toxicity to cells and tissues.

In this embodiment, the amount of active nitrogen species secreted from macrophages is reduced after treating the WHW extract of Example 1 with macrophages isolated directly from the abdominal cavity, RAW264.7 cells, which are mouse macrophages. We tried to check whether or not.

First, RAW264.7 cells and intraperitoneal macrophages were incubated for 24 hours, and then treated with various concentrations of WHW extract for 30 minutes. In addition, lipopolysaccharide (LPS) was treated to activate the cells to induce secretion of reactive nitrogen species. After culturing for 24 hours, the culture medium was collected and the concentration of activated nitrogen species secreted from the cells was measured by the Griess reaction method. We investigated how much the treated drug could inhibit the production of active nitrogen species, which are inflammation-inducing substances released from macrophages.

As a result, as shown in FIG. 2, the pharmaceutical composition of the present invention effectively reduced the production of excessively secreted activated nitrogen species from activated macrophages in both macrophages and macrophages isolated directly from the abdominal cavity of rats. Appeared.

② Inhibitory effect of inflammatory cytokine secretion

In general, when exposed to external pathogens, large amounts of cytokines, which mediate inflammatory reactions such as TNF-α or IL-1β, are released from macrophages to accelerate the inflammatory response. It is known to be closely associated with the development of various inflammatory diseases such as arthritis and degenerative brain diseases. In addition, an increase in inflammatory cytokines such as TNF-α has been reported to contribute to disease development by inducing cancer resistance as well as inducing insulin resistance in metabolic diseases such as cachexia, diabetes and obesity.

Thus, in the present Example, the WHW extract of Example 1 was treated with RAW264.7 cells, which are mouse macrophage lines and macrophages isolated directly from the abdominal cavity, and then excessively secreted from macrophages activated by external stimulation. The aim of this study was to determine whether TNF-α is effectively inhibited and the amount of TNF-α and IL-1β secreted from macrophages is decreased.

First, RAW264.7 cells and intraperitoneal macrophages were incubated for 24 hours and then treated with WHW extract at various concentrations for 30 minutes, and then treated with LPS to activate cells for 24 hours. The cell culture was collected and the concentration of cytokines secreted from the cells was measured by an enzyme-linked immunosorbent assay (ELISA). In this case, the ELISA kit for detecting each cytokine was purchased from eBioscience, and the cytokine concentration (pg / ml) in the culture medium was determined using a standard quantitative curve using recombinant proteins of each cytokine. (standard curve) was produced and then converted based on this.

As a result, as shown in FIG. 3, the WHW extract of the present invention was shown to strongly reduce the production of excessively secreted inflammatory cytokines from activated macrophages in proportion to the dose.

I. Inhibitory Effects of Kidney Fibrosis on Chronic Renal Failure

Progressive interstitial fibrosis usually leads to destruction of the tubules and capillaries, resulting in unstable diabetes (diabetes mellitus), hypertension, renal artery stenosis, kidney abnormalities and ureter obstruction ( Induced ureteral obstruction, especially when the growth factor (tumor growth factor) -β, which is a growth factor that promotes the growth and differentiation of blood vessels, is known to accelerate tissue fibrosis (renal fibrotic events).

In the case of chronic renal failure, renal cell destruction and fibrosis are accompanied, resulting in irreversible changes in the renal parenchyma, and the cells change into a myofibroblast-like phenotype. While the expression of TGF-β1 mRNA and α-SMA are increased in parallel, Smad2 (Sma- and Mad-) is a signal transducer that induces expression of the TGF-β1 mRNA and α-SMA. related protein 2) is known.

① TGF-β

Thus, in the present embodiment, TGF-β is induced to treat fibroblasts in MDCK cells (proximal tubule cell origin), which is a renal fibroblast cell line, and the WHW extract is treated by concentration according to the concentration of the WHW extract according to the present invention. The following experiment was conducted to determine whether it can be suppressed.

First, MDGF cells were treated with TGF-β (2.5 ng / ml) to induce renal fibrosis, and then treated with various concentrations of the WHW extract of the present invention for 24 hours, followed by 0.2% trypan blue staining method. Inhibition of renal cell fibrosis by extracts was examined by measuring cell viability.

As a result, as shown in Figure 4, when treated with MDG kidney cells 1mg / ㎖ of WHW extract of the present invention, renal fibrosis by inhibiting the cell viability of kidney cells caused by TGF-β treatment Appeared to be able to prevent.

② Smad2 (Sma- and Mad-related protein 2)

In addition, in the present embodiment, the following experimental method was performed to determine whether intracellular expression of Smad2, which is a signal transmitter of α-SMA and TGF-β, could be inhibited by the treatment of the WHW extract of the present invention.

First, MDGF cells were treated with TGF-β (2.5 ng / ml) and WHW extracts and cultured for 24 hours, and then the culture medium was removed and only cells were collected. In addition, RIPA buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% NP-40, 0.5% deoxycholic acid, 0.1% SDS) containing 1 mM PMSF and protease inhibitor cocktail was added to the collected cells. Protein was eluted. 30 ug of the extracted protein was separated by SDS-PAGE, and then transferred to a nitrocellulose membrane. After the reaction of Smad-2 antibody (anti-Smad-2 monoclonal antibody; mAb) and alkaline phosphatase conjugated mAb, the expression level of Smas-2 was measured by Chemiluminescence method.

As a result, as shown in Fig. 5, stimulating cells with TGF-β increases the gene expression of α-SMA and Smad in MDCK cells, and treatment of WHW extract effectively reduces their expression, resulting in fibrosis of kidney cells. Appeared to be able to prevent.

In view of the above results, the pharmaceutical composition according to the present invention can protect kidney cells by inhibiting the cytotoxicity and fibrosis induced by TGF-β, and gene expression of α-SMA and Smad, factors which accelerate fibrosis. It can be strongly suppressed.

In addition, the extract of the present invention was able to prove that the nephritis can be treated by preventing renal cell fibrosis in patients with chronic kidney failure.

All. Therapeutic Effect of Ischemic Acute Renal Ischemia

In general, reactive oxygen species (ROS) are known to cause various acute and chronic kidney diseases such as ischemic acute renal failure in the kidney, acute renal failure caused by antibiotics or toxic substances, and glomerulonephritis.

And all cells that consume oxygen under normal conditions convert 2-5% of the oxygen metabolized in the mitochondria into reactive oxygen groups such as superoxide and hydrogen peroxide (H ₂ O ₂ ). It has enzymes and substances that break down, and it has a mechanism to protect cells from its toxicity.

However, if the cell fails to perform normal protective mechanisms, the reactive oxygen groups left in the cell attack various components of the cell such as lipids, proteins, and DNA. Especially, in the case of the cell membrane of the cell, there are many unsaturated fatty acids to attack the reactive oxygen groups. Lipid peroxidation occurs easily and increases permeability of cell membranes, resulting in cell death resulting in cell death.

In addition, reactive oxygen groups are stimulated to secrete large amounts of inflammatory cytokines and fibroblast growth factors in the cell, thereby exacerbating nephritis through cell fibrosis.

Therefore, in this embodiment, after feeding the WHW extract of the present invention to the rat (renal ischemia / reperfusion mouse model) that induced ischemic acute renal failure, the degree of activity inhibition and reactive nephritis treatment of reactive oxygen groups was verified.

① Inhibitory effect of creatinine production

First, 10, 50, and 100 mg / kg of the WHW extract of the present invention was administered orally to a nephritis rat (Balb / c male mice, 8 week old) at 30 minutes and 1 day intervals for 14 days.

Then, blood was collected from the nephritis rats, and plasma was separated from the blood, and then creatinine levels were measured.

As a result, when nephritis is induced, the level of creatinine in plasma is increased, and as shown in FIG. 6, mice receiving the pharmaceutical composition according to the present invention have found that plasma creatinine is decreased.

② Inhibitory effect on kidney tissue deformation caused by nephritis

In general, when nephritis is induced, tubules are contracted and extracellular part of the interstitum is expanded to occupy the kidney, and the pharmaceutical composition of the present invention can be confirmed that it can reduce abnormal changes in the kidney. To this end, the morphological changes in the kidney tissues were analyzed by tissue staining, and the degree of contraction of tubules in the kidney tissues was analyzed by Masson trichrom staining.

First, kidneys extracted from rats were washed well with physiological saline, and then fixed with PLP (2% paraformaldehyde, 75 mM L-lysine, 10 mM sodium periodate) solution for one day. After embedding it again with paraffin solution, it was cut into 4㎛ thickness and attached to the slide to prepare tissue samples for staining. Tissue samples were stained using periodic acid shift (PAS) staining kit (DAKO) to observe the morphological changes of kidneys under a microscope.

In addition, the degree of contraction of the tubules in the renal tissue was measured by the Masson trichrom staining method of the number of tubules in the outer medular region. In other words, if there was no damage, grade 0, mild injury (circular endothelial cells, dilated tubular lumen) was grade 1, and moderate damage such as flat endothelial cells and expanded cells and tubules was grade 2 And a histological score dividing the severe damage characterized by endothelial cells flattened by the collapse of the nucleus, expanded tubules, and flattened into three grades.

As a result, as shown in Figure 7, the rat administered the pharmaceutical composition of the present invention, compared to the rat not fed the composition of the present invention was found to have less morphological deformation of the kidney tissue and swelling of the epileptic part, Tubular contraction and damage also decreased. At this time, Figure 7 A is a photograph showing a change in the shape of the kidney tissue, Figure 7 B is a measure of the degree of shrinkage of the tubules in the kidney tissue.

③ Inhibitory Effect of Active Oxygen Enzymes on Kidney of Nephritis Rat

In general, the generation of reactive oxygen groups is known to destroy kidneys by inducing oxidative stress in the cells to promote fibrosis of the kidneys.

Thus, in the present embodiment, catalase, a enzyme that is popular for the production of reactive oxygen species by fractionating cytoplasm and mitochondria of kidney cells to confirm whether the pharmaceutical composition of the present invention can protect the kidney of rat nephritis from ROS ), CuZnSOD (copper-zinc superoxide dismutase) and MuSOD (manganase superoxide dismutase) activity was measured.

As a result, as shown in Figure 8, when the nephritis rat was fed the WHW extract of the present invention (ischemia / reperfusion), the catalase (A), CuZnSOD (B) and MnSOD (C) activity by the WHW extract of the present invention All appeared to fall.

In other words, it was found that the WHW extract of the present invention protects kidney cells from renal cell fibrosis by inhibiting the activity of active oxygen enzymes involved in active oxygen production and consequently inhibiting the production of reactive oxygen species.

④ inhibitory effect on hydrogen peroxide and fatty peroxidation

As described above, if the production of reactive oxygen groups in the cell increases, destruction of the cell membrane structure by peroxidation or lipid oxidation, DNA damage by hydroxyl group (OH-), and DNA double bond structure or oxidation reaction including SH Degeneration of proteins has been a cause of kidney cell death (apoptosis).

Thus, in the present embodiment, after administering the pharmaceutical composition of the present invention to a nephritis rat, it was confirmed how it affects the peroxidation or lipid oxidation due to the generation of free radicals.

First, the kidneys extracted from the rats were put into a sucrose buffer (0.32M sucrose, 10mM Tris-HCl, pH 7.4) and crushed with a homogenizer, followed by centrifugation at 1,000 g for 5 minutes to recover the supernatant. The supernatant was again centrifuged at 15,000g for 30 minutes to collect cytosolic fractions, and then catalase activity and copper-zinc superoxide dismutase (CuZnSOD) activity were measured. In addition, the mitochondrial fraction obtained by recovering the supernatant and recovering the remaining precipitate was suspended in PBS added with 0.1% Triton X-100 and then crushed in a sonicator. The activity of CuZnSOD was measured from the supernatant obtained by centrifugation for 15 minutes at 15,000 g again. In other words, the activity of catalase and CuZnSOD was examined by measuring the absorbance due to the decomposition of hydrogen peroxide at 240 nm wavelength in mitochondria and cytosolic fraction, and SOD activity was measured using pyrogallol assay kit. .

In addition, the amount of hydrogen peroxide in the kidney was measured using xylenol orange, an iron-sensitive dye. In other words, hydrogen peroxide is oxidized by iron (II) and iron (III) in the presence of sorbitol, resulting in a complex form with the xyleneol origin of iron (III).

Lipid peroxidation in the kidney was measured by using a spectrophotometric assay to determine the amount of MDA (malodialdehyde) using thiobarbituric acid-reactive substances (TNARS).

As a result, as shown in Figure 9, the normal rats did not change the degree of peroxidation and lipid oxidation in the kidney cells regardless of administration of the composition of the present invention, but in the case of nephritis rats fatty acidization and peroxidation It has been shown to be inhibited by the composition of the present invention.

⑤ Expression effect of heat shock protein

In addition, oxidative stress, such as hydrogen peroxide injury or glucose deprivation, or cytotoxicity caused by glutamate may be caused by the expression of a heat shock protein (HSP-70). It is known to protect cells.

Thus, in this example, the composition of the present invention was administered to nephritis rats, and the expression level of HSP expressed in kidney cells was measured.

First, the kidneys extracted from rats were ground with a homogenizer, and then the expression level of HSP-70 was examined by Western blot analysis. That is, the protein obtained from the ground kidney tissue was separated by SDS-PAGE method and transferred to nitrocellulose membrane, and the membrane was treated with anti-HSP-27, anti-HSP-72 antibody and alkaline phosphatase-conjugated secondary antibody. Immunostaining. The expression of HSP-70 in the kidney cells was examined by reacting it with an ELC kit and exposing it to an X-ray film.

Result of this. As shown in FIG. 10, when the composition of the present invention was administered to nephritis-induced rats, the expression level of HSP was increased and the cells were protected from stress such as peroxidation and fatty acidization.

Example 4 Preparation of Enteric Extract

As described above, the Chinese herbal medicine extracts, which are preferably eluted from the duodenum and small intestine, were prepared for extract of the enteric medicinal extract.

end. material

Salvia 150g, Half 100g, Yellow Ginseng 60g, Sewage 60g, Yin Yang Guk 160g, Rhubarb 100g, Age 80g

I. Extraction process

Into the prepared enteric mixed herbal medicine 10 times the volume of the total medicinal herb volume of water, and heated at 100 ℃ for 3 hours to extract the hot water, the extract obtained by filtering the extract with gauze inlet temperature 120 ℃, extract flow rate 2 The enteric extract of the present invention was prepared by spray drying under a condition of ml / min and atomizing air pressure of 10 kPa.

Example 5. Preparation of Gastric Extract

As described above, it is preferable to select the herbal medicines that are preferably eluted from the above-mentioned herbal medicine extracts for which the prevention and treatment of renal failure has been confirmed.

end. material

Ginseng 150g, Leaflet 100g, Licorice 50g, Injin 300g, Taxa 200g, Baekbokyeong 80g, Baekchul 80g, Gyeji 40g

I. Extraction process

The total volume of herbal medicines 10 times the volume of the herbal medicine mixture prepared in the above-mentioned gastric medicinal herbs, heated at 100 ℃ for 3 hours to extract the hot water, and the extract obtained by filtering the extract with gauze, the inlet temperature 120 ℃, extract flow rate 2 The enteric extract of the present invention was prepared by spray drying under a condition of ml / min and atomizing air pressure of 10 kPa.

Example 6 Preparation of Double Nucleation Tablets

Using the enteric and gastrointestinal extracts prepared in Examples 4 and 5, the enteric extract was prepared as a double nucleus tablet so that the gastric extract was eluted in the duodenum and small intestine.

end. Furtherance

-Double core tablet (in 810mg tablet): Enteric tablet 210mg, stomach tablet 600mg

Enteric core tablets (in 210 mg of tablets): enteric extract 123.5 mg, microcrystalline cellulose 102 30 mg, colloidal silicon oxide 2.1 mg, corn starch 12.4 mg, crospovidone 6.2 mg, pregelatinized starch 15.2 mg, glyceryl behenate 10.3mg, Eudragit L100 6.3mg, Triethylcitrate 0.6mg, Talc 3.1mg

-Gastrointestinal external tablet (out of 600mg per tablet): Gastrointestinal extract 176.0mg, microcrystalline cellulose 102 312.0mg, colloidal silicon oxide 5.0mg, corn starch 30.0mg, crospovidone 15.0mg, pregelatinized starch 37.0mg, glyceryl Behenate 25.0mg

I. Manufacturing process

5% enteric coating was carried out after mixing, tableting (diameter 8mm, and hardness 4kp) at the enteric core tablet composition ratio. Enteric coated nuclear tablets and gastric external tablets were compressed according to a conventional method (diameter 12.5 mm, hardness 8 kp) to prepare a double tablet having a total weight of 810 mg.

Comparative Example 1. Preparation of Enteric Core Tablets

end. Furtherance

Enteric core tablets (in 210 mg of tablets): enteric extract 123.5 mg, microcrystalline cellulose 102 30 mg, colloidal silicon oxide 2.1 mg, corn starch 12.4 mg, crospovidone 6.2 mg, pregelatinized starch 15.2 mg, glyceryl behenate 10.3mg, Eudragit L100 6.3mg, Triethylcitrate 0.6mg, Talc 3.1mg

I. Manufacturing process

After mixing the materials weighed in the ratio was subjected to a tableting process (diameter 8mm, hardness 8kp) and then coated with 5% Eudragit L100 to prepare a enteric core tablet with a total weight of 210mg.

Comparative Example 2. Preparation of stomach tablets

end. Furtherance

Gastric external tablets (in 600 mg of tablets): Gastric extract 176.0 mg, microcrystalline cellulose 102 312.0 mg, colloidal silicon oxide 5.0 mg, corn starch 30.0 mg, crospovidone 15.0 mg, pregelatinized starch 37.0 mg, glyceryl behe Nate 25.0mg

I. Manufacturing process

After mixing the materials weighed in the ratio was compressed into tablets (diameter 11mm, hardness 8kp) to prepare a total external tablet of 600mg weight.

Example 7 Dissolution Experiment

For each tablet prepared in Comparative Examples 1, 2 and Example 3, using a solution having a pH similar to the gastric juice, duodenum and small intestine that is actually taken when taken by a real person, it is determined whether the solution is eluted with pH and time. It was.

To provide conditions similar to the actual human digestive process, first eluted in artificial gastric juice at pH 1.2 for 2 hours, followed by 2 hours in artificial duodenum at pH 6.8 and 6 hours in artificial small intestine at pH 7.4. Primary elution was performed. Icarin, which is a component of yin and yang, and cinnamic acid, which is a component of gage, among enteric materials, were analyzed using high performance liquid chromatography.

-Icarin Analysis Condition

Detector: UV absorbance photometer (wavelength 214 nm)

Column: inner diameter 4.6 mm, length 150 mm, octadecyl silica gel

Column Temperature: Room Temperature

Mobile phase: Acetonitrile water mixture (35.2: 64.8)

Flow rate: 1.0 mL / min

-Analysis of Sinnamsan

Detector: UV absorbance photometer (wavelength 280 nm)

Column: inner diameter 4.6 mm, length 150 mm, octadecyl silica gel

Column temperature: room temperature

Mobile phase: Acetic acid solution (1 → 15), acetonitrile mixture (56.4: 43.6)

Flow rate: 1.0 mL / min

As a result, as shown in FIGS. 11 and 12, in the case of gastric external tablets of Comparative Example 2, it was confirmed that all amounts of cinnamic acid, which is one of the gastric agents, were eluted over 90 minutes in artificial gastric juice of pH 1.2. In the enteric core tablet of Example 1, a small amount of Icarin, one of the enteric-materials, began to elute from the gastric juice of pH 1.2 starting at 60 minutes, and about 80% was eluted for 2 hours in the artificial duodenal solution of pH 6.8. It was confirmed that all remaining amounts were eluted from the artificial small intestine for 5 hours.

In addition, in the double core tablet of Example 6, cinnamic acid was eluted over 90 minutes in the artificial gastric juice of pH 1.2, and Icarin began to elute at 60 minutes, and then eluted for 5 hours in the artificial small intestine of pH 7.4. It became.

When the person actually taking the dual core tablet for preventing and treating renal failure of the present invention, the components of mansam, lobular, licorice, injin, taxa, baekbokyeong, baekchul, and gyejang, including cinnamic acid, are eluted to protect and strengthen gastrointestinal movement. In other words, through the duodenum and small intestine, Icarin and other types of salviae, Banja, Hwangyeon, Osuyu, Yin Yang Kwak, rhubarb, and herb are eluted, which means that the prevention and treatment of renal failure is maximized.

As described above, taking the double-nucleated tablet for preventing and treating renal failure of the present invention elutes ginseng, lobular, licorice, injin, taxa, baekbokyeong, baekchul, gyeji, including cinnamic acid to protect and enhance gastrointestinal movement. In the course of the duodenum and small intestine, Icarin, including Salvia, Banja, Huang Yan, Osuyu, Yin Yang Kwak, Rhubarb, and other ingredients are eluted to maximize the prevention and treatment of kidney failure.

1 is a graph showing the effect of reducing creatine following the administration of the composition of the present invention to patients with renal failure.

Figure 2 is a graph showing the effect of inhibiting the release of active nitrogen species from macrophages RAW264.7 cells (A) and intraperitoneal macrophages (B) after treatment of the composition of the present invention.

Figure 3 shows the effect of inhibiting the production of TNF-α (A, C) and IL-1β from macrophage lines RAW264.7 cells (A, B) and intraperitoneal macrophage lines (C, D) after treatment of the composition of the present invention The graph shown.

Figure 4 is a view showing the inhibitory effect of TNF-β on the treatment of the composition according to the invention in MDCK kidney cells.

Figure 5 shows the effect of inhibiting the expression of α-SMA (A) and Smad (B) after treatment with the composition of the present invention in TCK-stimulated MDCK kidney cells.

Figure 6 is a graph showing the effect of reducing creatinine in plasma for the composition of the present invention.

7 is a view showing the change in the shape of the kidney tissue (A) and the degree of tubular contraction (B) after treating the composition of the present invention for nephritis-induced kidney tissue.

8 is a composition showing the inhibitory effect of the active oxygen enzymes in kidney cells of nephritis rats of the present invention.

Figure 9 is a composition showing the inhibitory effect on the peroxidation (A) and lipid oxidation (B) in the kidney of rat nephritis rats.

Figure 10 is a view showing the effect of the composition of the present invention on the expression of heat shock protein peroxidation (A) and lipid oxidation (B) in kidney cells of nephritis rats.

FIG. 11 is a chromatogram showing the results of analysis using high performance liquid chromatography of Icarin or Cinnamic acid after 90 minutes or 9 hours after elution of enteric core tablets, gastric external tablets and double core tablets.

12 is a graph showing the elution% of Icarin or cinnamic acid with time after elution of enteric core tablets, gastric external tablets and double core tablets.

Claims (4)

Enteric core tablets containing mixed extracts of Chinese herbal medicine consisting of Salvia Radix, Panaxillas, Fructus, Fructus, Eumyanggok, Rhubarb, and Lung, as well as gastrointestinal protection and reinforcement as Gastric external tablets containing a mixed extract of the herbal medicine consisting of Taeksa, Baekbokryeong, Baekchul and Gyeji are tableted, but between the enteric core tablets and the stomach external tablets, enteric skin is coated, characterized in that the prevention and treatment of renal failure. The enteric core tablet according to claim 1, wherein when the total length of the enteric core tablet and the stomach external tablet is 100%, 6 to 10% by weight of salviae, 4 to 8% by weight, 3 to 5% by weight of yellow lotus, sesame oil Dual core tablet for renal failure prevention and treatment, characterized in that it comprises a mixed extract of the Chinese medicine consisting of 3 to 5% by weight, 8 to 12% by weight, yin and yang rhubarb 4 to 8% by weight and age 4 to 6% by weight. According to claim 1, wherein the stomach external tablets, 6 to 10% by weight of ginseng, 4 to 8% by weight lobular, 2 to 4% by weight licorice when the total amount of the enteric core tablet and the gastric external tablet is 100% Preventing and treating kidney failure, characterized in that it comprises a mixed extract of the Chinese medicine consisting of 15 to 20% by weight, 10 to 14% by weight, 4 to 6% by weight, 4 to 6% by weight, and 2 to 3% by weight of Baekjeung. Dual core tablet. The dual core tablet for preventing and treating renal failure according to any one of claims 1 to 3, wherein the extract is freeze-dried hot water extract of the herbal medicine.

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