Patent Description:
Currently, there are a total of <NUM> oral antiviral agents and <NUM> injection (peginterferon ex) as shown in Table <NUM> below for the treatment of chronic hepatitis B, and oral antiviral agents are divided into drugs with a high genetic barrier and drugs without, depending on the incidence of resistance.

Oral antiviral agents are nucleot(s)ide analogues that act on the process of converting pregenomic RNA of hepatitis B virus into DNA and interfere with normal DNA production, thereby inhibiting hepatitis B virus proliferation.

Peginterferon α, an injection, is known to have roles in destroying cccDNA and viral mRNA, inhibiting viral DNA replication, and regulating the immune response to virus-infected hepatocytes. However, the injection has a disadvantage in that the response rate for the therapeutic effect is low and that it cannot be used for people with reduced liver function due to the risk of liver failure. In addition, the use of peginterferon α should be considered carefully, as it can cause severe side effects such as depression. Combination therapy with peginterferon α and an oral antiviral agent has been suggested to be a more effective method for HBsAg loss, but the effect was not clear in genotype C, which accounts for the majority of Korean patients.

Therefore, domestic and international guidelines for chronic hepatitis B treatment recommend oral antiviral agents with a high genetic barrier to resistance as a first-line treatment, considering the advantages and disadvantages of each treatment.

Although oral antiviral agents can effectively inhibit the proliferation of hepatitis B virus, it is known that HBsAg loss, which is an index of a functional cure, rarely occurs in patients with chronic hepatitis B because they are not therapeutic agents that completely remove cccDNA in the nucleus.

When confirming the results of major clinical trials of oral antiviral agents, in the case of HBeAg-negative patients, there was no HBsAg loss after <NUM> year of treatment, as shown in Table <NUM> below (Non-patent reference <NUM>, <NPL>).

In the case of HBeAg-positive patients, the HBsAg loss rate after <NUM> year of treatment was only <NUM>% (TDF) at most, as shown in Table <NUM> below.

HBeAg loss and seroconversion, and HBsAg loss (or reduction) and seroconversion rate, which are the efficacy evaluation items of oral antiviral agents known to date, are as follows. It can be seen that it is difficult to achieve HBsAg loss with oral antiviral agents through the following contents.

The results of the second year of the phase <NUM> clinical trial for TAF (nucleotide family), the most recently approved therapeutic agent for chronic hepatitis B, have been announced (Non-patent reference <NUM>, Tenofoviralafenamide versus tenofovirdisoproxil fumarate for the treatment of patients with HBeAg-negative chronic hepatitis B virus infection: a randomised, double-blind, phase <NUM>, non-inferiority trial (<NPL>); Non-patent reference <NUM>, Tenofoviralafenamide versus tenofovirdisoproxil fumarate for the treatment of HBeAg-positive chronic hepatitis B virus infection: a randomised, double-blind, phase <NUM>, non-inferiority trial (<NPL>); Non-patent reference <NUM>, <NUM> weeks treatment of tenofoviralafenamide vs. tenofovirdisoproxil fumarate for hepatitis B virus infection (<NPL>)). The selection criteria for the phase <NUM> clinical trials of TAF included both patients with antiviral treatment experience and patients receiving treatment for the first time.

As shown in Table <NUM>, the HBeAg loss rate was <NUM>% at the first year and <NUM>% at the second year in the TAF group, and <NUM>% at the first year and <NUM>% at the second year in the TDF group, which were lower than those in the TAF group. In addition, the HBsAg loss rate was very low at around <NUM>% in both groups at both year <NUM> and year <NUM>.

In the case of TDF (nucleotide family), which has the largest share in the chronic hepatitis B treatment market, even the <NUM>-year follow-up results of phase <NUM> clinical trials have been announced (Non-patent reference <NUM>, <NPL>); Non-patent reference <NUM>, <NPL>); Non-patent reference <NUM>, <NPL>); Non-patent reference <NUM>, <NPL>. The main selection criteria for the phase <NUM> clinical trial were chronic hepatitis B patients who had been taking nucleot(s)ide-based antiviral drugs for less than <NUM> weeks. In the HBeAg-negative group, up to <NUM> out of <NUM> patients who had been taking Lamivudine or Emtricitabine (nucleoside family) for more than <NUM> weeks were allowed to enroll. In addition, HBeAg positive and HBeAg negative were classified and proceeded, and AdefovirDipivoxil (nucleotide family) was used as an active control group.

As shown in Table <NUM>, for the HBeAg loss rate, there was no mention of the result at the first year, and the rate was <NUM>% at the third year, <NUM>% at the seventh year, and <NUM>% at the tenth year. The HBeAg seroconversion rate was <NUM>% (TDF group) at the first year, <NUM>% at the third year, <NUM>% at the seventh year, and <NUM>% at the tenth year, respectively. The HBsAg loss rate in the HBeAg-positive group was <NUM>% at the first year (TDF group), <NUM>% at the third year (Kaplan-Meier analysis), <NUM>% at the seventh year (Kaplan-Meier analysis), and <NUM>% at the tenth year. In the HBeAg-negative group, only one patient had HBsAg loss until the fifth year, and the HBsAg loss rate was <NUM>% at the tenth year. Above all, it is worth noting that the loss of HBsAg did not appear in Asians (There was no mention of race in the results at the seventh year, but there were no Asians with HBsAg loss in the results at the tenth year. Kaplan-meier analysis, which was used to analyze HBsAg loss at the third and seventh years, is a method used for survival analysis in anticancer drug clinical trials to estimate the probability of survival over a period of time using death outcomes during the observation period. During the long-term follow-up study, patients with HBsAg loss and seroconversion dropped out or discontinued antiviral treatment, resulting in a loss of HBsAg loss subjects at the time of efficacy evaluation. To correct for this, Kaplan-meier analysis was used for follow-up study results at the third and seventh year of the TDF trial.

As a result of the efficacy evaluation at the first year, <NUM>% of patients in the HBeAg-positive group had HBeAg seroconversion, <NUM>% of patients had HBsAg loss, and <NUM>% of patients had HBsAg seroconversion.

Among the subjects who completed the phase <NUM> clinical trial, <NUM> patients participated in the long-term follow-up study, and <NUM> patients completed the <NUM>-year follow-up study. In addition, the patients assigned to the active control group also participated in the study by replacing the antiviral agent with TDF. As a result of evaluating the efficacy of the <NUM>-year follow-up study, <NUM>% of patients in the HBeAg-positive group showed HBeAg loss.

HBsAg loss occurred in <NUM> patients over <NUM> years, and all of them were HBeAg positive group subjects and non-Asians. Of these, <NUM> stopped taking antiviral agents.

Seven-year follow-up was completed for a total of <NUM> patients. The HBeAg loss rate was <NUM>%, and <NUM> patients showed HBsAg loss over <NUM> years, and <NUM> of them stopped taking antiviral agents. Only <NUM> of <NUM> patients belonged to the HBeAg-negative group.

Ten-year follow-up was completed for a total of <NUM> patients. The HBeAg seroanalysis results could be confirmed in only <NUM> patients, and as a result, HBeAg loss was found in <NUM>% of patients. In addition, HBsAg loss was also observed in <NUM>% (<NUM>/<NUM>) of patients.

Furthermore, the results of the TDF phase <NUM> clinical trial confirmed that not only HBsAg loss, but also reduction of HBsAg quantitative levels is not easy. In the TDF phase <NUM> clinical trial, patients who started receiving oral antiviral agent treatment for the first time showed a slight decrease in HBsAg levels at the beginning of treatment, but the degree and speed of the decrease slowed after <NUM> year of taking the antiviral agent, and after <NUM> years (<NUM> weeks), there has been no significant change since then (see <FIG>). This is a result confirming that long-term administration of oral antiviral agents does not cause a continuous reduction of HBsAg.

In the case of TDF (nucleotide family), which has the largest share in the chronic hepatitis B treatment market, even the <NUM>-year follow-up results of phase <NUM> clinical trials have been announced.

The results of phase <NUM> clinical trials for Entecavir, which had the No. <NUM> market share before TDF development, were announced up to the <NUM>-year follow-up results (Non-patent reference <NUM>, <NPL>); Non-patent reference <NUM>, <NPL>)). The phase <NUM> clinical trial for Entecavir was conducted on patients who had no experience taking nucleosidebased antiviral agents. Like the clinical trials for other antiviral drugs, the clinical trials were conducted by dividing HBeAg positive and HBeAg negative, but in the case of the HBeAg negative group, the results of HBeAg loss or HBsAg loss were rarely mentioned in the papers.

As shown in Table <NUM>, the HBeAg seroconversion rate was <NUM>% at the second year, and the seroconversion subjects did not participate in the <NUM>-year long-term follow-up study because they corresponded to the treatment response in the protocol. The HBeAg seroconversion rate in the <NUM>-year follow-up was <NUM>%. The HBsAg loss rate was <NUM>% at the second year and <NUM>% at the fifth year. In the case of Entecavir, apart from the phase <NUM> clinical trial, a seven-year real-world cohort study was conducted in Hong Kong for Chinese. As a result, the HBeAg seroconversion rate for <NUM> years analyzed by Kaplan-meier analysis was <NUM>%. HBsAg loss was seen in only <NUM> of <NUM> (<NUM>%) patients (Non-patent reference <NUM>,<NPL>)).

Through the summary of major clinical trial results for the three most commonly used drugs as the first-line treatment of chronic hepatitis B, it was confirmed once again that HBsAg loss, which means a functional cure for chronic hepatitis B, is difficult to occur with oral antiviral agents. Patients with chronic hepatitis B, who are difficult to cure, must take oral antiviral agents for almost the rest of their lives to suppress the virus. If these patients stop taking antiviral agents without being completely cured, the hepatitis B virus proliferation becomes active again, so treatment cannot be stopped in the middle.

However, long-term use of antiviral agents causes other problems such as resistance development, or occurrence of side effects like abnormal renal function or bone metabolic disease, so patients with chronic hepatitis B are in difficulties under any circumstances. Therefore, it is necessary to develop an effective therapeutic agent that can rapidly lead to a complete cure of chronic hepatitis B.

Accordingly, the present inventors have performed a highly inventive effort for the purpose of completely curing chronic hepatitis B, which is currently impossible to cure. As a result, the inventors completed the present invention by confirming in clinical trials that co-administration of an oral antiviral agent and a therapeutic vaccine comprising a lipopeptide and a poly(I:C) adjuvant resulted in a significant escalation effect (i.e., a synergistic effect) in the treatment index of chronic hepatitis B in patients receiving the standard treatment of antiviral agents, compared to patients receiving the standard treatment of antiviral agents alone, resulting in a cure.

It is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of chronic hepatitis B that enables a complete cure due to a significant escalation effect (i.e., a synergistic effect) in the treatment index of chronic hepatitis B, a pharmaceutical combined formulation, a combined formulation kit, and a method for generating an immune response against chronic hepatitis B in a subject comprising a step of administering the same to the subject.

To achieve the above object, in an aspect of the present invention, the present invention provides a pharmaceutical combined formulation for the prevention or treatment of chronic hepatitis B comprising an antiviral agent for hepatitis B; and a therapeutic vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of chronic hepatitis B comprising an antiviral agent for hepatitis B; and a therapeutic vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant.

In another aspect of the present invention, the present invention provides a method for generating an immune response against chronic hepatitis B in a subject comprising a step of administering a pharmaceutical combined formulation for the prevention or treatment of chronic hepatitis B comprising an antiviral agent for hepatitis B; and a therapeutic vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant to the subject.

In another aspect of the present invention, the present invention provides a combined formulation kit for the prevention or treatment of chronic hepatitis B comprising a first component containing an antiviral agent for hepatitis B; and a second component containing a vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant.

When the pharmaceutical composition, the pharmaceutical combined formulation, and the combined formulation kit provided in one aspect of the present invention are administered/used in hepatitis B patients, a remarkable synergy occurs in terms of therapeutic index for chronic hepatitis B, compared to patients who have undergone standard therapy including the administration of conventional antiviral agents, exhibiting the possibility of completely curing the disease.

The embodiments of this invention can be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below. It is well understood by those in the art who has the average knowledge on this field that the embodiments of the present invention are given to explain the present invention more precisely.

In addition, the "inclusion" of an element throughout the specification does not exclude other elements, but may include other elements, unless specifically stated otherwise.

In an aspect of the present invention, the present invention provides a pharmaceutical combined formulation for the prevention or treatment of chronic hepatitis B comprising an antiviral agent for hepatitis B; and a vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant.

At this time, the antiviral agent for hepatitis B can be an oral antiviral agent, and as a specific example of the oral antiviral agent, Entecavir, Tenofovirdisoproxil fumarate (TDF), Tenofoviralafenamide fumarate (TAF), Besifovirdipivoxil maleate, Lamivudine, Telbivudine, Clevudine, Adefovirdipivoxil, and the like can be used alone or in combination of two or more without limitation.

Hereinafter, the development trend of a new chronic hepatitis B treatment is described.

Since it is difficult to cure (functional cure) chronic hepatitis B with existing oral antiviral agents, the development of a new chronic hepatitis B treatment is actively underway. Recently, direct-acting antiviral agents with a new mechanism have been developed, and the representative types include RNA interferences that inhibit protein synthesis by binding to viral mRNA and breaking it down into small units, or capsid inhibitors that prevent normal production of a capsid covering genetic materials. On the other hand, immunotherapeutic agents of strategies with strategies to reduce HBsAg by activating weakened immune cells of patients are being developed, and the representative types include cytokines, tol-like receptor ligands that activate innate immunity, and therapeutic vaccines that control adaptive immunity by administering antigens. Development trends and types of chronic hepatitis B therapeutic agents are shown in Table <NUM> in the specification of the present invention.

Regarding the current status of clinical trials of newly developed chronic hepatitis B treatments, almost all clinical trials are being conducted with a design that confirms the treatment effect of the combination administration of clinical trial drugs while continuing to take antiviral agents, which are standard treatments. This is to protect patients by combining standard treatment, and because it was confirmed that it is difficult to completely cure chronic hepatitis B with only one type of treatment through clinical trials of oral antiviral agents or combined administration studies of oral antiviral agents and interferon. The development phase of chronic hepatitis B treatment and whether or not to administrate antiviral agents in combination are summarized in Table <NUM> below.

On the other hand, since the combied formulation is for the prevention or treatment of chronic hepatitis B, the antigen is preferably the entire surface antigen (L-HBsAg) of HBV (Hepatitis B Virus).

In addition, the adjuvant is a substance or a combination of substances that increases or induces an immune response to a vaccine antigen in a desirable direction in order to enhance the clinical effect of a vaccine when used together with a vaccine antigen. The main function of an adjuvant is to enhance and improve the clinical effectiveness of a vaccine, such as increasing and regulating an immune response to a vaccine antigen or extending the duration of the protective effect by acting on direct or indirect immune stimulation and antigen delivery. When a pathogenic bacterum or virus is infected, the surface receptor (pattern recognition receptor) of an immune cell recognizes the unique pattern (pathogen-associated molecular pattern, PAMP) of a pathogenic microorganism and causes an innate immune response. Toll like receptors (TLRs) are representative surface receptors, and about <NUM> types are known in humans. TLR ligands, which respond to tall-like receptors, are being developed as adjuvants because they directly stimulate immune cells to activate innate immune responses and induce humoral immunity and cellular immune responses, the acquired immunity against vaccine antigens, to protect the human body from infectious agents or contribute to tissue recovery.

Lipopeptide, a specific example of the adjuvant, was first synthesized by J. Metzger et al as a synthetic analogue of a lipopeptide derived from bacteria and mycoplasma (<NPL>). The molecular structure of the compound represented by the following formula (<NUM>) is N-palmitoyl-S-[<NUM>,<NUM>-bis(palmitoyloxy)-(2RS)-propyl]-[R]-cystein-SKKKK (pam3Cys-SKKKK), and various other analogues have been synthesized. <CHM>
<CHM>.

According to H. Schild et al. , when Pam3Cys-Ser-Ser was combined with an influenza virus T cell epitope and administered to mice, virus-specific cytotoxic T lymphocytes (CTLs) were induced. In general, lipopeptides are known as ligands for TLR2. The use of such lipopeptides is not limited to Pam3Cys-SKKKK, and a lipopeptide can consist of a fatty acid bound to a glycerol molecule and several amino acids. Specific examples thereof include PHC-SKKKK, Ole2PamCys-SKKKK, Pam2Cys-SKKKK, PamCys(Pam)-SKKKK, Ole2Cys-SKKKK, Myr2Cys-SKKKK, PamDhc-SKKKK, PamCSKKKK, Dhc-SKKKK, and the like. The number of fatty acids in a molecule can be one or more. The number of amino acids in a lipopeptide can be one or more. In addition, the fatty acid and amino acid can be chemically modified. Furthermore, the lipopeptide can be a lipoprotein, either as a part of a molecule or as a whole molecule, derived from a gram-positive or gram-negative bacteria or mycoplasma.

In addition, the poly(I:C) has been used as a potent inducer of type <NUM> interferon in in vitro and in vivo studies. Moreover, poly(I:C) is known to stably and maturely form dendritic cells, the most potent antigen-presenting cells in mammals (<NPL>). According to these previous reports, poly(I:C) is a potent IL-<NUM> inducer, and IL-<NUM> is an important cytokine that induces cellular immune response and formation of IgG2a or IgG2b antibody by driving the immune response to develop Th1. In addition, poly(I:C) is known to have strong adjuvant activity against peptide antigens (<NPL>). The poly (I:C) can have a length in a range of <NUM> to <NUM>,<NUM> bp, preferably <NUM> to <NUM>,<NUM> bp, and more preferably <NUM> to <NUM> bp, but not always limited thereto.

The lipopeptide and poly(I:C) can be included in the vaccine composition at a weight ratio of <NUM> to <NUM>:<NUM>, a weight ratio of <NUM> to <NUM>:<NUM>, a weight ratio of <NUM> to <NUM>:<NUM>, or a weight ratio of <NUM>:<NUM>, but not always limited thereto. However, the ratio can be adjusted to an appropriate level according to the patient's condition. In addition, the vaccine composition can be an aqueous solution formulation.

The vaccine composition can further include at least one selected from the group consisting of pharmaceutically acceptable carriers, diluents and adjuvants. For example, the vaccine composition can include a pharmaceutically acceptable carrier, and can be formulated for human or veterinary use and administered through various routes. The vaccine composition may be administered through oral, intraperitoneal, intravenous, intramuscular, subcutaneous, and intradermal routes. Preferably, it is formulated and administered as an injection. Injections can be prepared using aqueous solvents such as physiological saline and Ringer's solution, vegetable oils, higher fatty acid esters (e.g., ethyl oleate, etc.), and non-aqueous solvents such as alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.) and can include pharmaceutical carriers such as stabilizers (e.g., ascorbic acid, sodium sulfite, sodium pyrosulfate, BHA, tocopherol, EDTA, etc.) to prevent deterioration, emulsifiers, buffers for pH control, preservatives for preventing microbial development (e.g., chimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.) and preservatives (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.) to inhibit microbial growth. The vaccine composition can be administered in a pharmaceutically effective amount.

At this time, the term "pharmaceutically effective amount" means an amount sufficient to exhibit a vaccine effect but an amount not to cause side effects or serious or excessive immune responses. The exact dosage concentration depends on the antigen to be administered, and can be easily determined by those skilled in the art according to factors well known in the medical field, such as the patient's age, weight, health, gender, sensitivity to drugs, administration route, and administration method. The composition of the present invention can be administered once or several times. In addition, the adjuvant can be further included in the vaccine composition as other known adjuvants other than lipopeptide and poly(I:C).

In another aspect of the present invention, the present invention provides a pharmaceutical composition for the prevention or treatment of chronic hepatitis B comprising an antiviral agent for hepatitis B; and a vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant.

In another aspect of the present invention, the present invention provides a method for generating an immune response against chronic hepatitis B in a subject comprising a step of administering a pharmaceutical combined formulation for the prevention or treatment of chronic hepatitis B comprising an antiviral agent for hepatitis B; and a vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant to the subject.

If the vaccine composition is administered to a human (patient), it can be administered in an amount effective to stimulate an immune response in vivo, for example, it can be administered to humans once or several times, and the dosage is <NUM>-<NUM> µg, more preferably <NUM>-<NUM> µg, but not always limited thereto.

In another aspect of the present invention, the present invention provides a method for preventing, ameliorating or treating chronic hepatitis B comprising a step of administering an antiviral agent for hepatitis B; and a vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant to a subject.

In another aspect of the present invention, the present invention provides a use of [an antiviral agent for hepatitis B; and a vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant] for the preparation of a medicament for preventing, ameliorating or treating chronic hepatitis B.

When the pharmaceutical composition, the pharmaceutical combined formulation, and the combined formulation kit provided in one aspect of the present invention are administered/used in hepatitis B patients, a remarkable synergy occurs in terms of therapeutic index for chronic hepatitis B, compared to patients who have undergone standard therapy including the administration of conventional antiviral agents, exhibiting the possibility of completely curing the disease. This is directly supported by clinical trials of examples and experimental examples described below.

Hereinafter, the present invention will be described in detail by the following preparative examples, examples and experimental examples.

However, the following preparative examples, examples and experimental examples are only for illustrating the present invention, and the contents of the present invention are not limited thereto.

Recently, there is a growing need for the development of immunotherapeutic agents that can cause virus reduction in the body by resolving the lowered immune tolerance of patients with chronic hepatitis B. The therapeutic vaccine CVI-HBV-<NUM> is composed of L-HBsAg, a third-generation antigen with excellent immunogenicity, and L-pampo [Pam3Cys-SKKKK + poly(I:C)], a powerful adjuvant. When this is administered in combination with an antiviral agent, a synergistic effect of suppressing hepatitis B virus proliferation, resolving immune tolerance in patients and activating T cells can be expected. The ultimate goal of developing CVI-HBV-<NUM> is to induce adaptive immunity against hepatitis B virus, leading to a rapid cure (see <FIG>). More specifically, <NUM> or <NUM> µg of poly(I:C) was added to <NUM> or <NUM> µg of total surface antigen (L-HBsAg), mixed well, and then <NUM> or <NUM> µg of Pam3Cys-SKKKK, a lipopeptide, was added to the mixture to prepare the theapeutic vacine CVI-HBV-<NUM>.

A clinical trial was conducted in which an oral antiviral agent and the therapeutic vaccine CVI-HBV-<NUM> prepared above were co-administered to chronic hepatitis B patients who have undergone standard therapy including the administration of antiviral agents. In each experimental example described below, specific details such as the number of actually recruited patients, gender, age, and the type and dose of antiviral agents that have been prescribed are described. On the other hand, unless otherwise stated, patients with chronic hepatitis B were given antiviral agents through oral administration once a day, and the therapeutic vaccine CVI-HBV-<NUM> prepared above was administered by intramuscular injection three or six times every four weeks, or six times every two weeks together with an oral antiviral agent from the start of the combined administration in experimental examples described below.

The safety and tolerability of the therapeutic vaccine CVI-HBV-<NUM> for hepatitis B was confirmed through a <NUM>/2a-phase clinical trial in which an oral antiviral agent and the therapeutic vaccine CVI-HBV-<NUM> prepared above were co-administered to chronic hepatitis B patients who have undergone standard therapy including the administration of conventional antiviral agents.

A total of <NUM> patients were recruited for the <NUM>/2a clinical trial, of which <NUM> patients, excluding <NUM> patients who dropped out, were included in the efficacy evaluation analysis. Among them, <NUM> were male (<NUM>%) and <NUM> were female (<NUM>%), and their average age was <NUM> years old. The average length of time subjects took oral antiviral agents was approximately <NUM> months. As for the types of oral antiviral agents being taken, <NUM> patients took TDF alone, <NUM> patients took Entecavir alone, and <NUM> patients took Adefovir and other types of oral antiviral agents (Entecavir, Lamivudine or Telbivudine) together. Other than that, there were one patient taking TDF and Entecavir, one patient taking TDF and Telbivudine, one patient taking Lamivudine, one patient taking Telbivudine, and one patient taking three oral antiviral agents (Clevudine, Adefovir and TDF) together. They were orally administered once a day at a dose determined according to each antiviral agent they were taking (eq. Entecavir <NUM>, TDF <NUM>, Adefovir <NUM>).

The hepatitis B therapeutic vaccine CVI-HBV-<NUM> was administered with an antigen dose of <NUM> ug or <NUM> ug <NUM> times every <NUM> weeks, <NUM> times every <NUM> weeks, or <NUM> times every <NUM> weeks, depending on the group to which the subjects were assigned.

<FIG> is a diagram showing the results of analyzing the <NUM>/2a effectiveness of CVI-HBV-<NUM>.

As shown in <FIG>, it was confirmed that HBV-specific T cell immune response was induced in <NUM> of <NUM> patients (<NUM>%) after vaccine administration, resulting in overcoming immune tolerance by vaccine administration. In addition, a very encouraging result was obtained in which HBeAg serum loss was induced in <NUM> out of <NUM> (<NUM>%) subjects who maintained HBeAg-positive status even after taking antiviral drugs for a long time, and <NUM> out of <NUM> patients (<NUM>%) showed a quantitative decrease in serum HBsAg. In <FIG>, the results mentioned above are highlighted in red. From these results, it can be confirmed that HBV-specific T cells were activated by the administration of CVI-HBV-<NUM>, which greatly affected the quantitative reduction of HBsAg and the loss of HBeAg.

In order to compensate for the absence of a control group that took only antiviral agents in the <NUM>/2a clinical trial, the control group results of the phase <NUM> clinical trial of Gilead's therapeutic vaccine (GS4774) were compared (Randomized phase II study of GS-<NUM> as a therapeutic vaccine in virally suppressed patients with chronic hepatitis B (<NPL>)).

The results are shown in Tables <NUM> and <NUM> below.

As shown in Table <NUM>, Gilead's phase <NUM> clinical trial failed because it did not show a significant difference from the control group in the HBsAg change rate suggested as the primary endpoint, and the HBeAg loss rate in the GS4774 administration group was only <NUM>%, which was significantly different from the HBeAg loss rate of <NUM>% in the CVI-HBV-<NUM><NUM>/2a clinical trial.

As shown in Table <NUM>, as a result of comparison with the results of the CVI-HBV-<NUM> clinical trial using the results of the control group of the phase <NUM> clinical trial of GS4774, the control group of the phase <NUM> clinical trial of GS4774 did not show HBeAg loss in the <NUM>-year evaluation, which was a significant difference from the HBeAg loss rate of <NUM>% in the CVI-HBV-<NUM><NUM>/2a clinical trial. In addition, in the case of HBsAg reduction, comparison was made using the <NUM>/2a results analyzed at the same evaluation time point as the efficacy evaluation time point of the GS4774 clinical trial. As a result, it was confirmed that the amount of HBsAg reduction was greater in the <NUM>/2a clinical trial than in the control group of the GS4774 phase <NUM> clinical trial.

Through the comparison with the results of the GS4774 clinical trial, it was confirmed that it is difficult to overcome immune tolerance in chronic hepatitis B patients only by taking oral antiviral agents, and that the combined administration of CVI-HBV-<NUM> is more effective in HBeAg loss or HBeAg reduction.

Second, the results of the phase <NUM> clinical trial (<NUM> year) of TAF were compared with the results of the <NUM>/2a clinical trial of CVI-HBV-<NUM>. In the phase <NUM> clinical trial of TAF, TDF was used as an active control group, and the detailed results of the clinical trial are the same as the results of the phase <NUM> clinical trial of Tenofoviralafenamide (TAF) described above in the description o the related art [Table <NUM>]. The TAF phase <NUM> clinical trial results (including active control TDF results) were used to compare the <NUM>/2a efficacy results of CVI-HBV-<NUM> because the subjects were the most similar to the <NUM>/2a clinical trial of CVI-HBV-<NUM> among oral antiviral agent clinical trials, including patients with existing antiviral agent treatment experience in the TAF phase <NUM> clinical trial. In the phase <NUM> clinical trial of TAF, the proportion of subjects who had antiviral treatment experience was about <NUM>% to <NUM>%, although it varied slightly by group.

The results are shown in <FIG> and Table <NUM>.

<FIG> is a graph showing the results of comparing the HBeAg loss rate in the CVI-HBV-<NUM><NUM>/2a clinical trial and the TAF phase <NUM> clinical trial.

As shown in <FIG> and Table <NUM>, when comparing efficacy, CVI-HBV-<NUM> showed the highest HBeAg loss rate of <NUM>%, followed by TAF of <NUM>% and TDF of <NUM>%. Considering that the efficacy evaluation time points of the phase <NUM>/2a clinical trial were week <NUM>, week <NUM>, and week <NUM> significantly ahead of the TAF clinical trial evaluation time point (Week <NUM>, <NUM> year) compared to the baseline time point depending on the group, the above results were very encouraging and showed a synergistic effect of the combined administration of CVI-HBV-<NUM>. In addition, no subjects showed HBsAg loss in the CVI-HBV-<NUM> group, the HBsAg loss rate was less than <NUM>% in the TAF group, and only <NUM> out of <NUM> patients showed HBsAg loss in the TDF group. Meanwhile, HBV-specific T cell response was not included in the evaluation items in the oral antiviral agent clinical trial, so it could not be compared.

A long-term follow-up study was conducted to confirm the efficacy of the hepatitis B therapeutic vaccine (CVI-HBV-<NUM>) in subjects who participated in the phase <NUM>/2a clinical trial, and a total of <NUM> patients participated in this study.

As shown in Table <NUM>, as a result of quantitative analysis of HBsAg, it was confirmed that the HBsAg level was additionally reduced in <NUM> patients, <NUM> of which showed the HBsAg reduction rate of <NUM>% or more, and <NUM> patients showed the HBsAg reduction rate of <NUM>% or more. As a result of analyzing HBeAg serum loss, two additional patients showed HBeAg serum loss compared to the results at <NUM> weeks after the last CVI-HBV-<NUM> administration in the <NUM>/2a clinical trial.

Next, when the HBV-specific T cell immune response was analyzed, <NUM> out of <NUM> subjects (<NUM>%) showed the formation of more IFN-γ spots compared to the <NUM>/2a clinical trial baseline, confirming that T-cell activation was continuously maintained (see <FIG>). Among them, <NUM> (<NUM>%) patients showed the formation of more than twice as many IFN-γ spots as the baseline. Through these results, it was confirmed that the immune activation by the therapeutic vaccine CVI-HBV-<NUM> continued for a long time, and the possibility as a treatment for chronic hepatitis B was confirmed again.

The results of a long-term follow-up study conducted about <NUM> years after the subjects enrolled in the <NUM>/2a phase clinical trial were compared with the results of the second-year efficacy evaluation of the TAF phase <NUM> clinical trial. The results are shown in <FIG> and Table <NUM>.

<FIG> is a graph showing the results of comparing the HBeAg loss rate in the CVI-HBV-<NUM> long-term follow-up study and the TAF phase <NUM> clinical trial.

As shown in <FIG> and Table <NUM>, as a result of comparing the results of the long-term follow-up study conducted after an average of <NUM> years after subjects enrolled in the phase <NUM>/2a clinical trial and the second year efficacy evaluation result of the phase <NUM> clinical trial of TAF, the HBeAg loss rate after CVI-HBV-<NUM> administration was still the highest.

Regarding the HBeAg loss rate, the HBeAg loss rate in the CVI-HBV-<NUM> group was <NUM>%, <NUM>% in the TAF group, and <NUM>% in the TDF group.

To compensate for the absence of a control group in the <NUM>/2a clinical trial using CVI-HBV-<NUM>, other clinical trials that can be replaced with a possible control group were investigated and the efficacy was compared. As a result, it was confirmed that the combined administration of CVI-HBV-<NUM> was more effective than the administration of antiviral drug alone. In the clinical trial of Gilead's therapeutic vaccine (GS4774), no patients showed HBeAg loss in the group adminisyered with an antiviral agent alone, and the degree of T cell activation was lower than that of the group administered with GS4774, whereas <NUM>% of patients showed HBeAg loss, and <NUM>% of patients showed T-cell activation after CVI-HBV-<NUM> administration. These results confirm that CVI-HBV-<NUM> overcomes immune tolerance in chronic hepatitis B patients.

Claim 1:
A pharmaceutical combined formulation for use in the prevention or treatment of chronic hepatitis B comprising an antiviral agent for hepatitis B; and a vaccine composition including an antigen, a lipopeptide and a poly(I:C) adjuvant, wherein the antiviral agent for hepatitis B is an oral antiviral agent, wherein the oral antiviral agent is at least one selected from the group consisting of Entecavir, Tenofovirdisoproxil fumarate (TDF), Tenofoviralafenamide fumarate (TAF), Besifovirdipivoxil maleate, Lamivudine, Telbivudine, Clevudine and Adefovirdipivoxil, wherein the antigen is an entire surface antigen (L-HBsAg) of HBV (Hepatitis B Virus), and wherein the lipopeptide is at least one selected from the group consisting of Pam3Cys-SKKKK, PHC-SKKKK, Ole2PamCys-SKKKK, Pam2Cys-SKKKK, PamCys(Pam)-SKKKK, Ole2Cys-SKKKK, Myr2Cys-SKKKK, PamDhc-SKKKK, PamCSKKKK and Dhc-SKKKK.