KR101259688B1 - Pharmaceutical anti-herpesviral composition comprising angelicin - Google Patents

Abstract

The present invention provides a pharmaceutical composition for anti-herpes virus containing angelicin (angelicin) or a pharmaceutically acceptable salt thereof as an active ingredient. According to the present invention described above there is an advantage that can be efficiently treated or prevented diseases or diseases caused by the herpes virus, such as gamma herpes virus without drug toxicity or drug resistance.

Description

Pharmaceutical anti-herpesviral composition comprising angelicin}

The present invention relates to a pharmaceutical composition for anti-herpes virus containing angelicin as an active ingredient. More specifically, the present invention relates to a pharmaceutical composition for anti-herpes virus containing angelicin as an active ingredient capable of effectively treating or preventing diseases or diseases caused by the herpes virus such as gamma herpes virus.

Most human herpes viruses are 70-80% (See Table 1. Characteristics of human chronic viruses). It is a typical chronic virus that maintains infection for a lifetime by repeating latent infection and reactivation in the host.

Human herpes virus Kinds Latent infection A major disease HHV-1 Herpes simplex virus-1 Alpha Nervous system Herpes simplex, encephalitis HHV-2 Herpes simplex virus-2 Alpha Nervous system Genital herpes (STD), encephalitis HHV-3 Varicells-Zoster virus Alpha Nervous system Chickenpox, shingles HHV-4 Epstein-Barr virus gamma B lymph system Infectious mononucleosis, Bucket lymphoma, Nasopharyngeal carcinoma, B lymph tumor, Gastric cancer HHV-5 Human cytomegalovirus beta Lymphatic system Infectious mononucleosis, congenital malformations, retinitis, blindness HHV-6 Human herpesvirus-6 beta T lymph system Rash, meningitis, infectious mononucleosis, blunt hepatitis, atypical lymphocytosis HHV-7 Human herpesvirus-7 beta T lymph system HHV-8 Kaposi's
sarcoma-associated herpesvirus
gamma
B lymphocyte
Kaposi's sarcoma, B lymphoma

Herpes viruses are a typical opportunistic infection Reactivation due to decreased immune function or congenital infection causes various diseases and it shows a fatal infection prognosis according to the immune status of the patient. Prevention of Varicells-Zoster Virus (VZV) There are no preventive or therapeutic vaccines against the seven human herpes viruses except the live vaccine.

Control of reactivated herpes virus infection due to immune degradation, such as organ transplant and hematopoietic stem cell transplant, HIV-1 infection, is becoming an important issue.

In addition, the immune decline in the elderly population due to persistent infections of the herpes virus (see review article: Koch et al, Cytomegalovirus infection: a driving force in human T cell immunosenescence, Ann NY Acad Sci 2007 1114: 23-35) It will be an important health and social burden in Korea, which is entering an aging society.

Among herpes viruses, gamma herpesvirus is known to cause latent infection with lymph cells, causing tumors, and lytic infection in epithelial cells and fibroblasts. Gamma-herpes viruses include Epstein-Barr Virus (hereinafter referred to as EBV) and Kaposi's sarcoma associated virus (hereinafter referred to as KSHV).

Normally, the herpes virus is characterized by a high infection rate. Especially, in EBV, more than 90% of the world's population, including Korea, is seropositive. EBV is associated with nasopharyngeal carcinoma (NPC), gastric carcinoma (GC), Burkitt's lymphoma BL, post-transplant B cell lymphomas and PTBL during chronic infection by latent infection. ), And Hodgkin's disease (HD), which is closely associated with the formation of multiple tumors (Roizman, 2007).

KSHV is the most recent human infectious herpesvirus found in 1994. Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL), a type of B cell lymphoma (PEL) or multicentric casts Castleman's disease) (Alkan et al, 1997; Huang et al, 1995; Liberman et al, 1990; Moore et al, 1994).

Most of the existing anti-herpesvirus preparations were developed targeting the viral DNA polymerase (DNA pol). Among them, acyclovir and its derivatives are specifically phosphorylated by thymidine kinase (hereinafter, referred to as 'TK') of the herpes virus, and then enter the viral genome, which is replicated by DNA pol. It is a drug that induces premature termination and has been used most in the last 40 years after development. In addition, drugs such as forcarnet, such as a pyrophosphate analogue, are commonly used.

While they are effective in the treatment of alpha-herpes viruses, their clinical potency is limited for early infections (eg, infectious mononucleosis) or reactivation infections of beta or gamma-herpes viruses. In particular, there is no antiviral agent that can be useful for the control of gamma herpesvirus, and thus it relies mainly on steroid-based drugs for the treatment of hypertension.

In addition, recently, in patients with immunodeficiency due to organ transplantation, hematopoietic stem cell transplantation, and HIV infection, drug toxicity is severe, and more importantly, the occurrence of drug-resistant mutations in these patients is increasing (Danve-Szatanek C et al. Surveillance network for herpes simplex virus resistance to antiviral drugs: 3-year follow- up.J Clin Microbiol. 2004 42 (1): 242-9 .; Limaye AP et al. Emergence of ganciclovir-resistant cytomegalovirus disease among recipients of solid- organ transplants.Lancet 2000; 356: 645-649; Marfori JE et al. Development of new cytomegalovirus UL97 and DNA polymerase mutations conferring drug resistance after valganciclovir therapy in allogeneic stem cell recipients.J Clin Virol 2007; 38: 120-125).

Most drug-resistant mutations are mutations in TK or viral DNA pols, and it is very urgent to develop antihernospiral drug candidates with novel targets and mechanisms.

The present invention has been made to solve the problems of the prior art, the present invention is to provide a pharmaceutical composition having anti- herpesvirus activity without drug toxicity and drug resistance.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a pharmaceutical composition for anti-herpes virus containing angelicin (angelicin) represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

The pharmaceutical compositions of the present invention are useful for the treatment of infections of the herpes viruses such as the HSV-1, HSV-2, VZV alpha-helper viruses and beta-herpes viruses such as HCMV. Is preferably useful for gamma herpesviruses in which no particular antiviral agent is present and more preferably for Epstein-Barr Virus (EBV) or Kaposi's sarcoma associated virus (KSHV).

Although not limited thereto, the composition according to the present invention is characterized by having a therapeutic effect on a disease selected from the group consisting of infectious mononucleosis, Burkitt's lymphoma, nasopharyngeal carcinoma, B lymphoma, gastric cancer, Kaposi's sarcoma and Blymphoma. .

Angelicin used in the present invention may be used to purchase a commercially purified compound or synthesized, but preferably can be used to separate from the extract of the plant containing the compound, especially golgol ( Psoralea corylifolia ), golgol extract The separation method from can be used as long as it can separate the angelicin from the bone bones. For example, a polar or non-polar solvent soluble extract of angelicin may be purified by fractionation and washing several times with distilled water or the like, followed by a conventional fractionation process (Harborne J.B. et al., Phytochemical methods: A guide to modern techniques of plant analysis., 3rd Ed., pp 6-7, 1998).

 Although not limited to this, it is preferable that the angelicin is contained at a concentration of 15 to 120 µg / ml. Within this range, antiviral efficacy can be effectively exerted without toxicity.

The pharmaceutically acceptable salts of the above-mentioned formulas include salts of hydroxy groups which may exist in the compounds of the formulas, unless otherwise indicated. Examples of the salts of the hydroxy group include salts of alkali metals such as lithium, sodium, potassium, And alkaline earth metal salts such as calcium. Preferably, physiologically acceptable salts with sodium, potassium, calcium and the like can be cited, and these salts can be produced through the production method or the production process of salts known in the art.

In addition, the angelicin of the present invention may be prepared with pharmaceutically acceptable non-toxic salts and solvates according to methods conventional in the art. As the pharmaceutically acceptable salt, acid addition salts formed by free acid are useful. The acid addition salt is prepared by a conventional method, for example, by dissolving the compound in an excess amount of an acid aqueous solution, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The molar amount of the compound and the acid or alcohol (e.g., glycol monomethyl ether) in water may be heated and then the mixture may be evaporated to dryness, or the precipitated salt may be subjected to suction filtration.

Pharmaceutically acceptable salts of angelicin of the invention include salts of acidic or basic groups which may be present in the angelicin of the invention, unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts, including, but not limited to, salts known in the art. It may be manufactured through a manufacturing method or a manufacturing process.

 In the composition of the present invention, preferably from 0.0001 to 90% by weight of the angelicin relative to the total weight of the composition. In the composition of the present invention, preferably, the composition may have a form of powder, granule, tablet, capsule, injection, cream, gel, patch, spray or ointment formulation. The pharmaceutical composition containing the angelicin of the present invention as an active ingredient may further include appropriate carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

The pharmaceutical composition containing the angelicin as an active ingredient may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., or sterile injectable solutions, respectively, according to a conventional method. have.

In addition, carriers, excipients and diluents that may be included in the composition containing angelicin as an active ingredient include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate , Gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, Or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the compound of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention is preferably administered at 0.0001 to l00 mg / kg, preferably at 0.001 to 10 mg / kg. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The compounds of the present invention can be administered by various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

Hereinafter, the present invention will be described in more detail.

The development of antiviral agents against human gamma herpes virus is a costly and time-consuming method because the effects of drug inhibition are observed by Western blot or real-time PCR after reactivating the latent B cell line. The reality is that it depends entirely on systems that require labor. The convenience and efficiency are so poor that they are not suitable for screening systems.

Mouse gamma herpes virus (MHV-68) among gamma herpes viruses is an animal model of human gamma herpes virus with high genetic homology, and can easily identify latent infection as well as active replication in laboratory cell lines.

The inventors of the reporter virus (BC-3G, Patent Application No. 10-2009-0056926) inserted with enhanced green fluorescence protein, GFP expression of the MHV-68 / EGFP virus as an effective antiviral agent We have a screening system.

The cell-based screening system was used to detect anti-Herpis virus activity in 116 natural compounds-derived single compounds. As a result, angelicin effectively inhibited virus growth.

Angelicin discovered by the present inventors using the above system is a single compound isolated from the natural botanical ' bosgolji ' ( Psoralea corylifolia ) as an isomer of psoralen (Formula 1). ).

Angelicin has been reported to inhibit mitosis under UVA light and inhibit the proliferation of retroviruses such as HIV, but the mechanism is unknown (G. Miolo et al. 1994). In addition, the biological function of angelicin (angelicin) has been reported that it can be used to treat thalassemia by promoting the proliferation of γ-globin mRNA of human red blood cells (I. Lampronti et al . 2009). In addition, a derivative of angelicin 6a (Formula 2) has recently been reported to have anti-influenza activity (Yeh et al, 2010).

However, the literature review of the known papers and patents did not report that angelicin exhibited anti-herpes virus function. Thus, the present invention suggests the function of angelicin as a new anti-herpes virus agent. In addition, it can further provide new antiviral agents for the control of gamma herpes virus infection in which no specific antiviral agents are present.

According to the present invention configured as described above, angelicin has a relatively low side effects and toxic expression rate as a natural product extract component, has the effect of treating a disease or disease caused by the herpes virus without bio-friendly and drug resistance.

In particular, the present invention can very efficiently treat or prevent various diseases or disorders caused by gamma herpes virus without an antiviral agent, which can be usefully used for the control of gamma herpes virus.

In addition, angelicin according to the present invention is a useful substance having a new concept of anti-herpes virus activity, and can be used as a concomitant agent that can significantly reduce the dose of an anti-herpes virus agent having toxicity and side effects without degrading the activity. It may be.

BRIEF DESCRIPTION OF THE DRAWINGS It is a figure which shows the result of MHV-68 / EGFP virus replication control by angelicin treatment by this invention.
Figure 2 is a view showing the results of verifying the antiviral efficacy of the mouse gamma herpes virus of angelicin according to the present invention.
Figure 3 of Angelicin according to the present invention It is a figure which shows the result of having analyzed the viral gene expression inhibition to MHV-68 / EGFP.
4 is an angelicin of the present invention The figure which shows the result of having analyzed the antiviral effect on KSHV.
5 is an angelicin of the present invention The figure which shows the result of having analyzed the antiviral effect on EBV.
Figure 6 is a view showing the results of toxicity analysis for BHK-21 cell line of angelicin according to the present invention.
7 is a diagram showing the results of analyzing the effects on the angelicin KSHV infected cell lines (BC-3G, BCBL-1), EBV infected cell lines (B95.8), non-infected cell lines (DG75) according to the present invention.
8 is a view showing the results of analyzing the antiviral action of angelicin according to the present invention.

Hereinafter, the present invention will be described in detail with reference to Examples. However, the following examples illustrate the present invention, and the scope of the present invention is not limited by the following examples.

Example 1. Screening of Natural Compounds Derived from Antiviral Efficacy Against Mouse Gamma Herpes Virus: Discovery of Angelicin

We searched for anti-Herpisvirus activity in 116 natural compounds-derived single compounds with our cell-based screening system (BC-3G, see patent application 10-2009-0056926). ) Effectively prevented the growth of the virus.

More specifically, the present inventors observed the effect of a single compound library isolated from natural products on the replication of MHV-68 / EGFP virus infected with BHK21 (baby hamster kidney fibroblast cells) cell line by fluorescence microscopy. Screening based on the expression level, angelicin was found to inhibit the replication of the reporter virus without severe cytotoxicity.

A virus-treated sample and a sample treated only with DMSO, which is a solvent of a single compound, were treated as a negative control without antiviral effect, and ganciclovir (GCV), an antiviral agent that has already been demonstrated as an antiviral agent of gamma herpes virus: 20 μg / ml) and a sample treated with phosphoanoic acid (PAA: 100 μg / ml), an inhibitor of DNA polymerase, were determined as a positive control to determine the antiviral effect. All compounds were treated in culture of BHK-21 cell line at 3 hours prior to virus infection (incubation for 1 hour) and then at cell culture.

As a result, as shown in Figure 1, the anti-viral effect of the angelicin was increased, through which it was confirmed that the angelicin has anti-gamma herpes virus activity. In other words, when the MHV-68 / EGFP virus was infected with BHK-21 cell line with MOI = 0.5, an angelicin-treated sample was compared with the virus infection alone or when DMSO, a solvent of angelicin, was treated. In EGFP expression was reduced. In addition, it was confirmed that GFP expression was reduced by the inhibition of virus replication in a concentration-dependent manner (15 µg / ml, 30 µg / ml, 60 µg / ml) by angelicin. GFP shows GFP expression signal due to MHV-68 / GFP replication. This confirmed that angelicin has a concentration-dependent anti-gamma herpes virus activity (Fig. 1).

Example 2 Effect of Angelicin with Antiviral Efficacy on Mouse Gamma Herpes Virus

1) In order to verify the effect of angelicin found by the screening of Example 1 MHV-68 / EGFP virus was infected with BHK21 cell line so that MOI was 0.5 in the presence of various concentrations of angelicin cells after 2 days incubation period Western blot comparing the amount of MHV-68 / EGFP genes ORF45 and M9 protein, the culture supernatant is infected by Vero cells (plaque assay) by a plaque assay The extent of infectious virion production was measured.

Figure 2 shows the analysis of MHV-68 / EGFP virus protein expression (A) and infectious virion production (B) reduction by angelicin treatment. Protein expression levels of ORF45 and M9, the Lytic genes of MHV-68, were investigated by antiviral agents PAA and GCV and angelicin. As a result, it was observed that the expression of protein was reduced when PAA, GCV and angelicin were treated. In particular, the virus levels of infectious virions decreased by angelicin treatment compared to DMSO (negative control) virus only sample and DMSO. ) And it was confirmed that the band thickness of a specific protein is reduced through western blot (FIG. 2). The supernatant was infected with Vero cells, and the titer of the virus was confirmed by PFU (Plaque forming unit) appearing per 100 µl of the supernatant. Treatment with angelicin was found to decrease concentration of infectious virion in a dose dependent manner. Angelicin was treated with 15 µg / ml, 30 µg / ml and 60 µg / ml, respectively, and 100 µg / ml PAA and 20 µg / ml GCV. This confirmed that angelicin has an antiviral effect on MHV-68.

2) Real-time PCR was performed on ORF56 sites of MHV-68 genomic DNA to determine whether angelicin affects viral genome replication. In order to determine the effect on the viral gene expression by comparing the virus transcripts, the progress of RT-PCR, RT-Q-PCR to analyze the gene expression degree difference was confirmed and the results are shown in FIG. The upper panel of FIG. 3 contains an ORF56 gene of MHV-68 by angelicin, PAA, and GCV per 50 ng of genomic DNA to determine the ratio in the genomic DNA of BHK-21 cells. The ORF56 was measured by real-time PCR and the relative amounts were compared based on the untreated samples. The lower panel of Figure 3 shows the result of comparing the amount of the transcript with the thickness of the band on the gel by proceeding the RT-PCR for the ORF50, ORF29 after separating the RNA to synthesize the cDNA to compare the difference in the amount of transcript . Relative expression of RTA, a representative early transcript of the virus, was compared by RT-Q-PCR. In both experiments, actin expression was used as a loading control.

As shown in FIG. 3, it was confirmed that the viral load or transcript production was reduced in the sample treated with angelicin as compared to the negative control. Compared with the samples treated with the positive control PAA and GCV, it was confirmed that the amount of viral genome was reduced and the amount of transcripts decreased for ORF50, ORF29, and RTA depending on the concentration of angelicin. Therefore, it was verified that angelicin inhibits DNA replication and reduces gene expression in the viral genome.

Example 3 Antiviral Effect of Angelicin on Human Gamma-2 Herpesvirus, KSHV

In order to confirm that the antiviral effect of angelicin identified in the model of mouse gamma herpes virus has the same effect on similar human gamma herpes virus, KSHV was progressed to BC-3G cell line and BCBL-1 cell line which were latent infected cells. KSHV and MHV-68 both belong to gamma-2 herpesvirus.

Figure 4 A shows the results of analysis by flow cytometry (angel flow cytometry) whether angelin can inhibit the ability to induce virus reactivation by TPA in BC-3G cells. B and C of Figure 4 shows the results confirmed by Western blot expression of the virus Lytic protein RTA. At this time, 1 hour after treatment with angelicin was treated with TPA (20ng / ㎖).

BC-3G obtained in this laboratory is a reporter cell line expressing GFP by lytic replication of the virus. When BC-3G was treated with angelicin and TPA, which is a substance that induces virus reactivation, the percentage of GFP expressing cells was confirmed, and it was confirmed that the rate of reactivation was reduced when angelicin was treated (FIG. 4A). ).

In order to prove this by molecular biological methods, Western blot was used to identify the difference in the expression level of RTA protein, an early gene of KSHV activator virus replication of BC-3G, a KSHV latent infection cell line. As shown in B of FIG. 4, it was confirmed that the amount of RTA expression decreased in the sample treated with angelicin as compared to the amount of protein expressed in the TPA treated sample.

The same effect was confirmed in BCBL-1, another KSHV + cell line, with the same results of decreased RTA protein expression when angelicin was treated with TPA (FIG. 4C). Through this, it was verified that angelicin has an antiviral effect of inhibiting the virus proliferation of human gamma herpes virus KSHV.

Example 4. Human Body Antiviral Effect of Angelicin on Gamma-1-Herpisvirus, EBV

Another clinically important human gamma herpesvirus and gamma-1 herpesvirus, EBV, were also confirmed to have antiviral effects.

Angelicin virus was induced by EBV-incubated B95.8, Raji, HR-1 cell lines treated with angelicin (60 or 120 μg / ml) for 1 hour before inducing virus reactivation with TPA (20 ng / ml). Western blot and immunofluorescence assay were performed to determine the expression level of EA-D, a representative lytic gene of EBV. In mouse gamma herpes virus or KSHV, The antiviral effect shown was the same (FIG. 5).

A, B and C of Figure 5 is analyzed by Western blot in the B95.8, Raji, HR-1 cell line by the expression of EA-D, the lytic protein of EBV by angelicin One result is shown, and D of FIG. 5 shows the result confirmed by immunofluorescence assay in Raji cell line.

Example 5 Cytotoxicity of Angelicin

1) Toxicity test on BHK-21 cells

To determine whether the antiviral effect of angelicin is toxic to cells, we performed Tryphan blue exclusion assay and MTT assay on BHK-21 cells capable of viral replication. More specifically, after treatment with angelicin, PAA, and GCV dissolved in DMSO in the BHK-21 cell line, a daily Trypan blue exclusion assay and MTT assay were performed from day 1 to day 3 to 100% of the untreated sample. After determination, relative cell growth was measured (FIG. 6). As a result, as shown in Figure 6, it was confirmed that no apparent cytotoxicity at a concentration of less than 120㎍ / ㎖ angeline. This confirmed that the antiviral efficacy of angelicin below 60 ㎍ / ㎖ concentration is not due to cytotoxicity.

2) Toxicity test on BC-3G, BCBL-1, B95.8 and B95.8 cell lines

MTT assay and Trypan blue exclusion assay were performed to determine whether anti-viral angelicin was cytotoxic to KSHV latent cell lines BC-3G and BCBL-1 cells and EBV latent cell line B95.8 cells. . In other words, after treatment with the angelicin extract dissolved in DMSO in BC-3G, BCBL-1, B95.8 cell line and B95.8 cell line, MTT assay was performed from day 1 to day 3 to measure no sample. Was set to 100% and the relative cell growth was measured (Fig. 7). The results of the Trypan blue exclusion assay did not represent the same results as the MTT assay. As a result, as shown in Fig. 7, the cytotoxicity of BC-3G and BCBL-1 cells and B95.8 cells was weak compared to the effect of angelicin on BHK-21 cell line. Not shown.

Example 6. Antiviral Mechanism of Angelicin

To determine which stages of the virus's life history have antiviral effects, pre-treatment with and pre-treatment with angelicin three hours prior to infection with the animal model of gamma herpes virus, MHV-68 After 1 hour after the virus was removed and replaced with a medium (media) containing normal FBS, the treatment was carried out (post treatment) to the treatment with angelicin. When all treated samples were referred to as All treatment, Western blot and plaque assays were performed with each pellet and supernatant to confirm the protein level of the virus and the degree of infectious virion production.

FIG. 8A shows the difference in viral protein expression between two samples, FIG. 8B shows the result of real-time PCR checking the amount of ORF56 in genomic DNA, and FIG. 8C shows the supernatant using The result of confirming the infectious virion production difference is shown, and FIG. 8D shows the result of confirming the expression level of the ORF50 gene by RNA extracted from the cell.

As shown in FIG. 8, the pretreatment-only sample appeared to be slightly different from the negative control, but the post-treatment only showed viral protein expression and infectious virion. Production, the amount of the viral genome in genomic DNA (Gnomic DNA), it was confirmed that the gene expression of the RTA is significantly reduced (Fig. 8). This means that the antiviral effect of inhibiting the function of the virus is better when the virus is treated with angelicin after infection than before the infection, and can be used as a treatment for gamma herpes virus infection.

As described above, gamma herpes virus infection usually causes lifelong infection in the lymphocytes of the host, and in immunocompromised patients due to old age or organ transplantation, a problem may be caused by reactivation of gamma herpes virus. . The development of antiviral agents of gamma herpes virus will be clinically important in terms of both preventing and preventing these diseases.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is obvious that the same is by way of illustration and example only and is not to be construed as limiting the scope of the invention. It is therefore intended that the scope of the present invention be defined by the appended claims and their equivalents.

Claims (8)

Epstein-Barr Virus (EBV) or Kaposi's sarcoma-associated herpesvirus (KSHV) containing angelicin represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient Pharmaceutical composition for anti-herpes virus having antiviral activity.
(Formula 1)

The pharmaceutical composition of claim 1, wherein the angelicin is derived from golgol ( Psoralea corylifolia ). delete delete delete delete The method of claim 1, wherein the composition is characterized in that it has a therapeutic effect on a disease selected from the group consisting of infectious mononucleosis, Burkitt's lymphoma, nasopharyngeal carcinoma, B lymphoma, gastric cancer, Hodgkin's lymphoma, Kaposi's sarcoma and primary exudative lymphoma. Pharmaceutical composition for anti- herpes virus. According to claim 1, wherein the angelicin is a pharmaceutical composition for anti- herpes virus, characterized in that it contains a concentration of 15 ~ 120 ㎍ / ㎖.

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