KR20030063308A - B-type infectious hepatitis medicine using of a phyllanthus and method of the same - Google Patents

Abstract

PURPOSE: A method of preparing an agent for treating Type B hepatitis using an extract of Phyllanthus ussuriensis or Phyllanthus urinaria is provided. By the administration of the Phyllanthus ussuriensis extract to a patient suffering from Type B hepatitis, HBV-DNA is detected to be negative within 3 months after the administration thereof and medical findings of transaminase have been improved. CONSTITUTION: An agent for treating type B hepatitis comprises an extract of Phyllanthus ussuriensis or Phyllanthus urinaria and an excipient containing microcrystalline cellulose, hydroxypropyl cellulose or magnesium stearate. Phyllanthus ussuriensis or Phyllanthus urinaria is washed with water, cut into 1 to 2cm, washed with purified water at 30 to 40deg.C and extracted with 20 times of purified water at 93deg.C for 10hr. Thereafter, the extract is filtered, concentrated at 45deg.C for 4hr in a vacuum concentrator, cooled to -70deg.C or less for 24hr and freeze-dried at -15 to -75deg.C after centrifugation.

Description

B-type INFECTIOUS HEPATITIS MEDICINE USING OF A PHYLLANTHUS AND METHOD OF THE SAME}

The present invention relates to a therapeutic agent for non-hepatitis comprising the extract of the pearl plant and a method for producing the same, more specifically, the aqueous extract of Phyllanthus (Phyllanthus) pearlescent clinically effective against hepatitis B in humans The present invention relates to a therapeutic agent for non-hepatitis comprising a extract of pearlescent plants obtained by preparing a pharmaceutical composition useful for the treatment of hepatitis B by adding various excipients to the treatment solution.

As is well known, hepatitis B virus (hereinafter abbreviated as HBV) is a virus of the Hepadnaviridae strain that is specifically infected with the human body, and has an incubation period of about 60 to 110 days and undergoes various degrees of clinical trial. 90-95% recover completely from infection. In patients who do not recover from infection, HBV DNA is assimilated into genomic DNA of human hepatocytes, leading to chronic active hepatitis, cirrhosis, liver cancer, and the like. Chronic hepatitis caused by HBV is a disease with a high mortality rate that causes chronic viral infections, lymphomas and chronic kidney disorders, and eventually leads to death.

Humans began to become infected with HBV in B.C. It is estimated to be around 2000, and there is already a description of jaundice in BC. The hepatitis B virus was discovered by Bloomberg (B.S.) in 1964, and it is also known that hepatitis B can cause liver cirrhosis and liver cancer. In Korea, Chung (W.K.) has studied the pathology and clinical aspects of hepatitis B viral hepatitis, and especially found a viral hepatitis that does not express jaundice.

Currently, about 200 million people worldwide are carriers of HBV, and 80% of them are concentrated in the Asia-Pacific region. In Korea, hepatitis B surface antigen (HBsAg) is reported in 7% of the population. (Lee Se-hoon et al., A Study on the Relationship between the Positive Rate of HBsAg and HBs and Liver Function Tests in Patients with Comprehensive Health Examination, Journal of the Korean Society of Immunology, 7: 265-273, 1965).

Despite the development of HBV vaccines since 1981 and immunizations for those who may be exposed to newborns and hepatitis, the number of carriers has not been significantly reduced to date. This is due to the vertical reduction of HBV (infection from the mother to the fetus). This carrier is the main cause of development (Chung, DK et al., Vertical transmission of hepatitis B antigen, J. Catholic Med. Coll., 27: 256-267, 1976; Chung, WK et al., Preventin of perinatal transmission of hepatitis B virus: Acomparison between the efficacy of passive and passive-active immunization in Korea, J. Infec. Dis., 151: 280-286, 1985). In the case of children born to HBeAg-positive persons, especially those who are born to HBeAg-positive, although infection through the placenta is not common, they may be immune to later HBV if exposed to HBV during birth or placenta damage or sensitized to HBeAg through placenta. It can cause defects and, as a result, it is more likely that newborns will become chronic carriers. Therefore, women who are already carriers of hepatitis B should be cured before their age of 20 at the latest, or at least HBeAg lost in their blood.

Recently, PCR (Polymerase Chain Reaction) technology has been introduced into this field, and it is now possible to measure HBV DNA in carriers' blood, so the time when HBeAg was the only indicator of hepatitis B has passed. In order to reduce the absolute number of hepatitis B carriers and to prevent the occurrence of hepatitis, liver cirrhosis and liver cancer, not only in Korea, but also in Asia-Pacific, HBV is given to newborns. There is a need to develop a method for significantly lowering the amount of DNA itself.

Although several drugs have been developed as a treatment for hepatitis B, few have satisfactory efficacy, and the fact that currently used drugs such as Ara-A and Acyclovir are also highly toxic. It is becoming. Phosphonoformate (Foscarnet), Suramin, Prostaglandin, 2-carbocyclic analogue of 2-deoxyguanosine, and Famciclovir are known to be promising. Much verification is required. To date, very few drugs have been used in humans to improve HBsAg, HBeAg, and HBV-DNA, which are indicators of hepatitis B without side effects, and even more drugs with fewer side effects are available for mothers or women of childbearing age. It's hard to see.

Benkateswaran et al. (1987), Bloomberg et al. (1988), and Yanagi (1989) used the Phyllanthus plant (Euphorbiaceae), which was used early in India, to treat hepatitis B. Was intended. According to Unander, DW et al., Journal of Ethnopharmacology, 34 (1991) 97-133), which summarized data from around the world on the use and biochemical analysis of Pilandus plants in 1991, Among the 550 species of Pilandus, such as jaundice or hepatitis, P. niruri, P. airy-shawii (amarus (?)), P. amarus, P. fraternus webster, P. urinaria, P. gasstroemi Muell-Arg and It consists of seven species including P. thymoides. P. amarus and P. niruri are described in Venkateswaran, PS et al., Effect of an extract from Phyllanthus niuri on hepatitis B and woodchuck hepatitisvirus: in vitro and in vivo studies, Proc. Natl. Acad. Sci , 84: 274, 1987) and hepatitis B virus in ducks (Nis, J., et al., Effect of Phyllanthus amarus on duck hepatitis B virus replification in vitro, J. of Med. Virol., 32: 212- 218, 1990), indicating its effectiveness but failing in actual clinical use (Leelarasamee, A., et al., Failure of Phyllanthus amarus to eradicate hepatitis B surface antigen from symptomless carriers, Lancet, 1: 335, 1990). In addition, P. thymoides had the most effective binding ability with HBsAg in vitro, but it was due to the specific lectin binding to HBsAg. (Shead, A. et al., Neutralization but not cure of duck hepatitis B by Australian Phyllanthus extracts.Abstract 602 In: Scientific Program and Abstracts volume, the 1990 International Symposium on Viral Hepatitis and Liver Disease, April 4-8, 1990, Houston , Texas). In view of the above, some examples of P. amarus, P. nururi, etc. among the plants of Pilandus have been used as folk remedies for jaundice or hepatitis, and they have suggested the effectiveness of hepatitis B in vitro. Indeed, the effectiveness of hepatitis B in humans has not been proven.

The present invention has been made in view of the above-described prior art, and concentrates an aqueous extract of Phyllanthus (Jinchocho) which is clinically effective against human hepatitis B, and injects various excipients into the treatment liquid. It is an object of the present invention to provide a therapeutic agent for non-hepatitis comprising a extract of pearlescent plants obtained by preparing a pharmaceutical composition useful for the treatment of hepatitis B and a method for producing the same.

1 is a view showing a manufacturing process of a therapeutic agent for hepatitis hepatitis comprising an extract of the pearl plant according to an embodiment of the present invention.

In order to achieve the above object, according to a preferred embodiment of the present invention, a therapeutic agent for hepatitis hepatitis comprising an extract of a pearl plant comprising an extract of Phyllanthus and an excipient of microcrystalline cellulose, hydroxypropyl cellulose, magnesium stearate Is provided.

On the other hand, the present invention is the process of washing each plant with water collected and cultivated phyllanthus (Phyllanthus) as a raw material by using a shredding cutter after washing each plant with water and air-dried at a constant temperature so that the stem is 1 to 2 Cm and; Washing and washing the leaves and stems again with purified water having a temperature of 30 ° C. to 40 ° C., and again washing twice with purified water once the mixed washing is completed; Adding purified water as much as 20 times the amount of Pearl Candle (Phyllanthus), and heating for 10 hours while maintaining the temperature of 93 ° C .; When the pearl herb (Phyllanthus) is extracted through heating with purified water, passing through a 400 mesh filter to yield 1100 L; Concentrating by using a vacuum concentrator to extract 200 liter / hr per hour while maintaining the concentration at 25Brix (yield: 40 liter) for 4 hours at a temperature of 45 ℃ once the pearl vinegar (Phyllanthus) is extracted completely; If the concentrate is pigmented, rapidly freezing the concentrate using a rapid freezer at a temperature of −70 ° C. for 24 hours; Centrifuging and extracting the extract and freeze-drying for 36 hours (yield 10-12Kg) at -15 ° C to -75 ° C; The freeze-dried extract is pulverized again through a grinder having a particle size of 16 mesh; Method for preparing a hepatitis therapeutic agent comprising an extract of pearl vinegar, characterized in that the excipient of microcrystalline cellulose, hydroxypropyl cellulose, magnesium stearate is added to the milled product and mixed in about 10 minutes. This is provided.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail with reference to drawings.

1 is a view showing a manufacturing process of a therapeutic agent for hepatitis hepatitis comprising an extract of the pearl plant according to an embodiment of the present invention.

[Gathering of pearl vinegar]

Jinju used in the present invention is an annual herb belonging to the Euphorbiaceae family, and the pearl candles are collected wild from the wild fields of Gyeongsangbuk-do and Jeju-do respectively, and their seeds are obtained and grown to obtain a test material. .

[Production of Hepatitis Treatment Agent Using Pearl Herbal Medicine]

First, each plant was collected or cultivated with Phyllanthus (Phyllanthus) was washed with water and air-dried at a constant temperature, and then chopped using a shredding cutter so that the stem is 1 to 2 Cm, 30 ℃ ~ 40 After washing the leaves and stems again with clean water at a temperature of ℃ ℃, and washed separately twice in purified water once the mixed wash is completed.

Then, 20 times more purified water was added to the input of Phyllanthus and heated for 10 hours while maintaining the temperature of 93 ° C. That is, 1200 liters of purified water is added per 60 kg of pearl candles in one batch.

When the pearl herb (Phyllanthus) is extracted through heating with purified water, it is passed through a 400 mesh filter to yield 1100 liters.

When the pearl plant (Phyllanthus) is extracted completely, it is concentrated using a vacuum concentrator to 200 L / hr per hour while maintaining the concentration at 25 Brix (40 L yield) for 4 hours at a temperature of 45 ℃ again.

When the concentrate is pigmented, the concentrate is rapidly frozen using a rapid freezer for 24 hours at a temperature of −70 ° C. or lower, and the extract is centrifuged and filtered, followed by 36 hours (−10 ° C. to −75 ° C.). 12 kg) lyophilized.

The lyophilized extract is pulverized again through a crusher having a particle size of 16 mesh, and mixed by adding an excipient to the pulverized product. The excipient is to mix microcrystalline cellulose, hydroxypropyl cellulose and magnesium stearate in about 10 minutes.

The material based on Phyllanthus thus prepared shows the ability to bind HBsAg in serum and hepatitis B vaccine in vitro. Due to this binding capacity, these plant extracts inhibit the binding of HBsAg and HBsAb, and each has an antagonistic inhibitory effect at a concentration of 1.25mg / ml.

In addition, the extract of Phyllanthus inhibits HBV DNA polymerase activity in vitro, and inhibits HBV DNA polymerase activity in proportion to concentration at about 34.1 μg / ml or more. In particular, weary extract (Phyllanthus) is effective in causing severe degeneration of the virus at 0.1% concentration.

Phyllanthus has also shown remarkable effects in several clinical trials: HBV-DNA was negative in serum and positive for HBsAb in serum when P. urinary extract was administered to healthy HBV carriers for 3 months. Came out (ELISA method). Even in cases where mild chronic hepatitis was suspected as a carrier of HBV, HBsAb was positive after 3 months of P. urinary administration.

However, HBsAb was negative when radioisotope was used, probably because a substance that specifically binds HBsAg appears in the blood when absorbed in Phyllanthus. In the case of hepatic cirrhosis combined with chronic active hepatitis due to HBV infection, HBV-DNA became negative only 3 months after administration of Phyllanthus, and the activities of GOT and GPT in serum were significantly improved.

In view of the various test results, it can be seen that the aqueous extract of Phyllanthus can be used as an effective hepatitis B treatment with little toxicity or side effects.

The hepatitis B therapeutic composition of the present invention comprises an aqueous extract of Phyllanthus as an active ingredient, and other conventionally acceptable pharmaceutically acceptable excipients such as carriers, diluents, fillers, anti-flocculation agents, lubricants, and flavoring agents. In addition, it can be prepared into pharmaceutical preparations such as oral, subcutaneous, and intravenous injections according to known methods. The hepatitis B therapeutic agent may be administered orally or parenterally, oral dosage may be used as an aqueous extract in an amount of 100 to 1,000 mg, preferably 200 to 600 mg per kg body weight per day. It can be adjusted according to various conditions such as patient's symptoms, age, sex, and weight.

(1) Test of binding capacity using HBsAg positive patient serum

The extract was dissolved in distilled water, and 400 μl of HBsAg positive serum was mixed with 400 μl of solution prepared at final concentrations of 10, 5, 2.5, 1.25 and 0.63 mg / ml, and reacted at 20 ° C. for 1 hour and centrifuged at 3000 rpm for 15 minutes. After separation, the titer of HBsAg in the upper layer was measured to evaluate the degree of reduction of HBsAg. The degree of inhibition of the binding of HBsAg and HBsAb in serum by Phyllanthus extract is shown in Table 1 below.

TABLE 1

plant Binding inhibition according to the concentration of plant extract (mg / ml) (%) 10 5 2.5 1.25 0.63 Pearl Candle (Phyllanthus) 98.9 92.5 92.7 90.8 72.0

[Efficacy test 4-in vitro]

[Electromicroscopic Observation of Phyllanthus Extracts on HBV]

The HBV particles were obtained by layering the serum of HBsAg and HBeAg positive patients on a 20% sucrose cushion and ultrafast centrifugation (Sorvall) for 4 hours at 4 ° C. and 100,000 g for 4 hours, and then pelleting the first with phosphate buffer (PBS). Suspended to 1/20 of the amount. 2% aqueous solution of Phyllanthus extract was diluted 10-fold and 20-fold again with distilled water (0.2 and 0.1% solution)

Determination of the inactivation of HBV of the phyllanthus extract was performed by distilled water as a control, mixing the diluted P. urinaria 0.2 and 0.1% solution with the same amount of HBV particle solution at room temperature for 30 minutes. After standing for a while, the morphological changes of HBV particles were observed by electron microscopy.

Electron microscopic observation showed that a drop of the prepared sample solution was dropped on the grid, negatively stained with 1% phosphotungstic acid, washed twice with PBS, and the excess moisture remaining on the grid was measured in Wattman No. As a result of thorough removal, the result of observing with a electron microscope (Zeiss) showed that the extract of Phyllanthus caused severe degeneration of the virus at 0.1% concentration.

[Hepatitis B Virus Carrier of Chronic Active Hepatitis with Liver Cirrhosis]

The liver function test showed GOT / GPT = 129/184, HBsAg (+) HBeAg (+) anti-HBcAb (+) anti-HBsAb (-), and there was no mass on the ultrasonography and there was a slight coarse shadow. . The physical findings were relatively healthy, but there were arachnoid angiomas and palmar hematomas in the chest, and hepatic cirrhosis of 2 cm below the right rib margin was found. Liver biopsy revealed that cirrhosis combined with chronic active hepatitis could not be applied to interferon.

To this patient, the change of serologic findings is shown in Table 2 below, with 200 mg / kg body weight per day of Phyllanthus extract.

TABLE 2

Time-lapse Indicators of HBV Infection Transaminase HBsAg HBsAb HBeAg HBeAb HBV / DNA GOT GPT Before treatment + - + - + 269 342 1 month after treatment + - + - + 143 242 3 months after treatment + (+) + - - 49 112

As shown in Table 2, HBV-DNA became negative and 3 months after the administration of the extract of Pearlflower (Phyllanthus) extract and the transaminase was also found to be very improved. However, due to cirrhosis of the liver, continuous observation is required.

As described above, it can be seen that the pearl leaf (Phyllanthus) extract can be used as an effective hepatitis B treatment with little toxicity or side effects.

On the other hand, non-hepatitis therapeutic agent and its preparation method comprising the extract of the pearl plant according to an embodiment of the present invention is not limited only to the above embodiments, various modifications can be made within the scope not departing from the technical gist. .

As described above, the treatment for non-hepatitis hepatitis comprising the extract of the pearlescent plant according to the present invention and a method for producing the same, HBV-DNA became negative only 3 months after the administration of the extract of Pearlflower (Phyllanthus) and the appearance of transaminase It can be seen that the improvement was very good. Therefore, the pearl leaf (Phyllanthus) extract has the effect that can be used as an effective hepatitis B treatment with little toxicity or side effects.

Claims (2)

A therapeutic agent for hepatitis hepatitis comprising an extract of a pearlescent plant comprising an extract of a phyllanthus and an excipient of microcrystalline cellulose, hydroxypropyl cellulose and magnesium stearate. Collecting or cultivating primrose, which is a primitive vinegar, by washing each plant with water, air-dried at a constant temperature, and slicing using a shredding cutter to make the stem 1 to 2 Cm; Washing and washing the leaves and stems again with purified water having a temperature of 30 ° C. to 40 ° C., and again washing twice with purified water once the mixed washing is completed; Adding purified water as much as 20 times the amount of Pearl Candle (Phyllanthus), and heating for 10 hours while maintaining the temperature of 93 ° C .; When the pearl herb (Phyllanthus) is extracted through heating with purified water, passing through a 400 mesh filter to yield 1100 L; Concentrating by using a vacuum concentrator to extract 200 liter / hr per hour while maintaining the concentration at 25Brix (yield: 40 liter) for 4 hours at a temperature of 45 ℃ once the pearl vinegar (Phyllanthus) is extracted completely; If the concentrate is pigmented, rapidly freezing the concentrate using a rapid freezer at a temperature of −70 ° C. for 24 hours; Centrifuging and extracting the extract and freeze-drying for 36 hours (yield 10-12Kg) at -15 ° C to -75 ° C; The freeze-dried extract is pulverized again through a grinder having a particle size of 16 mesh; Method for preparing a hepatitis therapeutic agent comprising extracts of pearlescent plants characterized in that the excipient of microcrystalline cellulose, hydroxypropyl cellulose, magnesium stearate is added to the ground product and mixed in about 10 minutes. .