A drug combination preparation for preventing or treating chronic hepatitis B, its use, and combination preparation kit

Abstract

The present invention relates to a pharmaceutical composition, a pharmaceutical combination preparation, and a combination preparation kit, each of which contains an oral antiviral drug as an active ingredient and a therapeutic vaccine containing a lipopeptide and a poly (I: C) adjuvant. When the pharmaceutical composition, the pharmaceutical combination preparation, and the combination preparation kit are administered / used in hepatitis B patients, a significant synergistic effect occurs in the therapeutic index of chronic hepatitis B, compared with patients who have received standard treatment including administration of conventional antiviral drugs, showing the possibility of completely curing the disease.

Description

Background of the Invention 1. Technical Field

[0002] The present invention relates to a pharmaceutical composition, a pharmaceutical combination preparation and a combination preparation kit for preventing or treating chronic hepatitis B, each of which comprises an oral antiviral drug and a therapeutic vaccine containing lipopeptide and poly (I: C) adjuvant as active ingredients.

[0003] 2. Description of Related Technology

[0004] Currently, as shown in Table 1 below, there are a total of 8 oral antiviral drugs and 1 injection (pegylated interferon α) for the treatment of chronic hepatitis B, and according to the incidence of drug resistance, oral antiviral drugs are divided into drugs with high genetic barriers and drugs without high genetic barriers.

[0005] [Table 1] Types of therapeutic agents used for chronic hepatitis B

[0006]

[0007] Oral antiviral drugs are nucleoside analogs that act on the process of converting the pregenomic RNA of the hepatitis B virus into DNA and interfere with normal DNA production, thereby inhibiting the proliferation of the hepatitis B virus.

[0008] The injection of pegylated interferon alpha is known to play a role in destroying cccDNA and viral mRNA, inhibiting viral DNA replication, and regulating the immune response to virus-infected hepatocytes. However, the injection has the disadvantage that the response rate to the therapeutic effect is low, and it cannot be used in people with reduced liver function due to the risk of liver failure. In addition, the use of pegylated interferon alpha should be carefully considered because it can cause serious side effects such as depression. Combination therapy of pegylated interferon alpha and oral antiviral drugs is considered to be a more effective method for HBsAg loss, but the effect is unclear in genotype C, which accounts for the majority of Korean patients.

[0009] Therefore, considering the advantages and disadvantages of each treatment method, domestic and international guidelines for the treatment of chronic hepatitis B recommend oral antiviral drugs with high genetic barriers to resistance as first-line treatment.

[0010] Although oral antiviral drugs can effectively inhibit the proliferation of HBV, HBsAg loss, which is known to be an indicator of functional cure, rarely occurs in chronic HBV patients because they are not therapeutic agents that completely remove cccDNA in the cell nucleus.

[0011] When the results of the main clinical trials of oral antiviral drugs were confirmed, in the case of HBeAg-negative patients, there was no HBsAg loss after 1 year of treatment, as shown in the following Table 2 (Non-patent Document 1, EASL 2017 Clinical Practice Guidelines on the management of hepatitis B virus infection).

[0012] [Table 2] Evaluation results of the efficacy of oral antiviral drugs in HBeAg(-) patients

[0013]

[0014] In the case of HBeAg-positive patients, the HBsAg loss rate after 1 year of treatment was only 3% at most (TDF), as shown in Table 3 below.

[0015] [Table 3] Evaluation results of the efficacy of oral antiviral drugs in HBeAg(+) patients

[0016]

[0017]

[0018] HBeAg loss and seroconversion as well as HBsAg loss (or reduction) and seroconversion rate are the efficacy evaluation items of oral antiviral drugs known so far, as shown below. It can be seen from the following content that HBsAg loss is difficult to achieve with oral antiviral drugs.

[0019] (1) Results of Phase 3 clinical trials of tenofovir alafenamide (TAF)

[0020] Results from the second year of phase 3 trials of the recently approved chronic hepatitis B therapeutic TAF (nucleotide family) have been published (Non-patent reference 2, Tenofoviralafenamide versus tenofovirdisoproxilfumarate for the treatment of patients with HBeAg-negative chronic hepatitis B virus infection: a randomised, double-blind, phase 3, non-inferiority trial (Lancet Gastroenterol Hepatol 2016; 1:196-206); Non-patent reference 3, Tenofoviralafenamide versus tenofovirdisoproxil fumarate for the treatment of HBeAg-positive chronic hepatitis B virus infection: a randomised, double-blind, phase 3, non-inferiority trial (Lancet Gastroenterol Hepatol 2016; 1:185-95); Non-patent reference 4, 96 weeks treatment of tenofoviralafenamide vs. tenofovir disoproximal fumarate for hepatitis B virus infection (Journal of Hepatology, 2018). The selection criteria for the phase 3 clinical trial of TAF include patients with experience in antiviral treatment and patients who are receiving treatment for the first time.

[0021] 1.1 Summary of Key Efficacy Results

[0022] [Table 4] Results of Phase 3 clinical trials of tenofovir alafenamide (TAF)

[0023]

[0024]

[0025] As shown in Table 4, the HBeAg loss rate in the TAF group was 14% in the first year and 22% in the second year, and the HBeAg loss rate in the TDF group was 12% in the first year and 18% in the second year, which were lower than those in the TAF group. In addition, the HBsAg loss rate in both groups was very low, about 1%, in the first and second years.

[0026] (2) Phase 3 clinical trial results and long-term follow-up efficacy results of tenofovir disoproxil fumarate (TDF)

[0027] For TDF (nucleotide family), which has the largest market share in the chronic hepatitis B treatment market, even the 10-year follow-up results of Phase 3 clinical trials have been published ((Non-patent reference 5, Tenofovir Disoproxil Fumarate versus Adefovir Dipivoxil for Chronic Hepatitis B (Nengl ​​j med 359; 23, 2008); Non-patent reference 6, Three-Year Efficacy and Safety of Tenofovir Disoproxil Fumarate Treatment for Chronic Hepatitis B (GASTROENTEROLOGY 2011; 140: 132-143); Non-patent reference 7, Seven-Year Efficacy and Safety of Treatment with Tenofovir Disoproxil Fumarate for Chronic Hepatitis B Virus Infection (Dig Dis Sci. 2015 May; 60(5): 1457-64); Non-patent reference 8, Ten-year efficacy and safety of tenofovir disoproxilfumarate treatment for chronic hepatitis B virus infection (Liver International. 2019; 00: 1-8.)). The main selection criteria for the Phase 3 clinical trial were patients with chronic hepatitis B who had been taking nucleoside-based antiviral drugs for less than 12 weeks. In the HBeAg-negative group, up to 120 of the 382 patients who had been taking lamivudine or emtricitabine (nucleoside family) for more than 12 weeks were allowed to participate. In addition, HBeAg-positive and HBeAg-negative were classified and treated, and adefovir dipivoxil (nucleotide family) was used as an active control group.

[0028] 2-1. Summary of key efficacy results

[0029] [Table 5] Phase 3 clinical trial results and long-term follow-up study efficacy results of tenofovir disoproxil fumarate (TDF)

[0030]

[0031]

[0032] As shown in Table 5, for the loss rate of HBeAg, there was no result mentioned in the first year, 34% in the third year, 54.5% in the seventh year, and 52.2% in the tenth year. The HBeAg seroconversion rate was 21% in the first year (TDF group), 26% in the third year, 39.6% in the seventh year, and 27.3% in the tenth year. The HBsAg loss rate in the HBeAg-positive group was 3.2% in the first year (TDF group), 8% in the third year (Kaplan-Meier analysis), 11.8% in the seventh year (Kaplan-Meier analysis), and 4.9% in the tenth year. In the HBeAg-negative group, only one patient lost HBsAg in the fifth year, and the loss rate of HBsAg in the tenth year was 3.4%. Most importantly, it is worth noting that there was no HBsAg loss in Asians (race was not mentioned in the seventh year results, but there were no Asians with HBsAg loss in the tenth year results). The Kaplan-Meier analysis method for analyzing HBsAg loss at the third and seventh years is a method used for survival analysis in clinical trials of anticancer drugs to estimate the probability of survival over a period of time using the outcome of death during the observation period. In long-term follow-up studies, patients with HBsAg loss and seroconversion withdraw or stop antiviral treatment, resulting in the loss of subjects with HBsAg loss during efficacy evaluation. To correct for this, the Kaplan-Meier analysis method was used for the third and seventh year follow-up study results of the TDF trial.

[0033] 2-2. Detailed efficacy results by year

[0034] a. Phase 3 clinical trial (1 year) results:

[0035] As a result of the first-year efficacy evaluation, 21% of patients in the HBeAg-positive group had HBeAg seroconversion, 3.2% of patients had HBsAg loss, and 1.3% of patients had HBsAg seroconversion.

[0036] b. 3-year follow-up results:

[0037] Of the subjects who completed the Phase 3 clinical trial, 585 patients participated in the long-term follow-up study and 542 patients completed the 3-year follow-up study. In addition, patients assigned to the active control group also participated in this study, where TDF was used instead of antiviral drugs. As a result of the efficacy evaluation of the 3-year follow-up study, 34% of patients in the HBeAg-positive group showed HBeAg loss.

[0038] Over the 3 years, 20 patients experienced HBsAg loss, all of whom were HBeAg-positive subjects and non-Asians, and 14 of them stopped taking antiviral drugs.

[0039] c. Results of 7-year follow-up:

[0040] A total of 437 patients completed seven years of follow-up. The HBeAg loss rate was 54.5%, and 28 patients experienced HBsAg loss over the seven years, of whom 25 stopped taking antiviral drugs. Only 1 of the 28 patients belonged to the HBeAg-negative group.

[0041] d.10-year follow-up results:

[0042] A total of 203 patients completed ten years of follow-up. HBeAg serum analysis results were confirmed in only 23 patients, thus, HBeAg loss was found in 52.2% of patients. In addition, HBsAg loss was also observed in 4.0% (8 / 198) of patients.

[0043] 2-3. Declining trend of HBsAg level

[0044] In addition, the results of the TDF Phase 3 clinical trial confirmed that not only HBsAg is lost, but also the reduction of HBsAg quantitative levels is not easy. In the TDF Phase 3 clinical trial, patients who started oral antiviral treatment for the first time showed a slight reduction in HBsAg levels at the beginning of treatment, but the degree and speed of the reduction slowed down after taking antiviral drugs for 1 year, and after 2 years (96 weeks), there was no significant change thereafter (see Figure 1). This result confirms that long-term oral antiviral drugs do not cause a continuous reduction in HBsAg.

[0045] (3) Phase 3 clinical trial results of entecavir and efficacy results of long-term follow-up studies

[0046] In the case of TDF (nucleotide family), which has the largest market share in the treatment of chronic hepatitis B, even the 10-year follow-up results of the Phase 3 clinical trial have been announced.

[0047] Entecavir ranked first in market share before the development of TDF, and its Phase 3 clinical trial results have been published, including 5-year follow-up results ((Non-patent reference 9, Entecavir Therapy for up to 96 Weeks in Patients With HBeAg-Positive Chronic Hepatitis B (GASTROENTEROLOGY 2007; 133: 1437-1444); Non-patent reference 10, Entecavir Treatment for up to 5 Years in Patients with Hepatitis B e Antigen-Positive Chronic Hepatitis B (HEPATOLOGY, Vol. 51, No. 2, 2010)). The Phase 3 clinical trial of entecavir was conducted on patients who had no experience taking nucleoside-based antiviral drugs. Like clinical trials of other antiviral drugs, clinical trials were conducted by dividing HBeAg positive and HBeAg negative, but in the case of the HBeAg negative group, HBeAg loss or HBsAg loss results were rarely mentioned in the literature.

[0048] [Table 6] Phase 3 clinical trial results of entecavir and efficacy results of long-term follow-up studies

[0049]

[0050]

[0051] As shown in Table 6, the HBeAg seroconversion rate in the second year was 31%, and the seroconversion subjects did not participate in the 5-year long-term follow-up study because they corresponded to the treatment response in the scheme. The HBeAg seroconversion rate in the 5-year follow-up was 23%. The HBsAg loss rate was 5% in the second year and 1.4% in the fifth year. For entecavir, in addition to the Phase 3 clinical trial, a seven-year real-world cohort study of the Chinese population was conducted in Hong Kong. As a result, the 7-year HBeAg seroconversion rate was 82.1% by Kaplan-Meier analysis. HBsAg loss was observed in only 4 (2.5%) of the 160 patients (Non-patent reference 11, Seven-Year Treatment Outcome of Entecavir in a Real-World Cohort: Effects on Clinical Parameters, HBsAg and HBcrAg Levels (Clinical and Translational Gastroenterology, 2017)).

[0052] (4) Summary of clinical trial results of oral antiviral drugs: limitations of monotherapy

[0053] By summarizing the results of major clinical trials using the three most commonly used drugs as first-line treatment for chronic hepatitis B, it is confirmed once again that oral antiviral drugs are difficult to cause HBsAg loss, and HBsAg loss means functional cure of chronic hepatitis B. Patients with chronic hepatitis B who are difficult to cure must take oral antiviral drugs to suppress the virus for almost the rest of their lives. If these patients stop taking antiviral drugs without complete cure, the hepatitis B virus proliferation will become active again, so treatment cannot be stopped in the middle.

[0054] However, long-term use of antiviral drugs causes other problems such as the development of drug resistance, or the occurrence of side effects such as abnormal renal function or bone metabolic diseases, so patients with chronic hepatitis B have difficulties in any case. Therefore, it is necessary to develop an effective therapeutic agent that can quickly and completely cure chronic hepatitis B.

[0055] Therefore, the present inventors have made highly creative efforts to completely cure chronic hepatitis B, which is currently impossible to cure. As a result, the present inventors have completed the present invention by demonstrating in a clinical trial that the combined administration of an oral antiviral drug and a therapeutic vaccine containing a lipopeptide and a poly (I: C) adjuvant results in a significant incremental effect (i.e., a synergistic effect) on the therapeutic index of chronic hepatitis B in patients receiving standard treatment with antiviral drugs, compared with patients receiving standard treatment with antiviral drugs alone, thereby leading to cure. Summary of the invention

[0056] The object of the present invention is to provide a pharmaceutical composition for preventing or treating chronic hepatitis B (which can completely cure chronic hepatitis B due to the significant incremental effect (i.e., synergistic effect) of the therapeutic index of chronic hepatitis B), a pharmaceutical combination preparation, a combination preparation kit and a method for generating an immune response against chronic hepatitis B in a subject, the method comprising the step of administering the pharmaceutical composition, pharmaceutical combination preparation, and combination preparation kit to the subject.

[0057] To achieve the above objectives, in one aspect of the present invention, the present invention provides a pharmaceutical combination preparation for preventing or treating chronic hepatitis B, which comprises an antiviral drug for hepatitis B; and a therapeutic vaccine composition containing an antigen, a lipopeptide and a poly (I: C) adjuvant.

[0058] In another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating chronic hepatitis B, which comprises an antiviral drug for hepatitis B; and a therapeutic vaccine composition containing an antigen, a lipopeptide and a poly (I: C) adjuvant.

[0059] In another aspect of the present invention, the present invention provides a method for producing an immune response against chronic hepatitis B in a subject, the method comprising the step of administering a drug combination preparation for preventing or treating chronic hepatitis B, the drug combination preparation comprising an antiviral drug for hepatitis B; and a therapeutic vaccine composition comprising an antigen, a lipopeptide and a poly (I:C) adjuvant.

[0060] In another aspect of the present invention, the present invention provides a combination preparation kit for preventing or treating chronic hepatitis B, which comprises a first component containing a hepatitis B antiviral drug; and a second component containing a vaccine composition, which vaccine composition comprises an antigen, a lipopeptide and a poly (I: C) adjuvant.

[0061] Beneficial Effects

[0062] When the pharmaceutical composition, pharmaceutical combination preparation and combination preparation kit provided by one aspect of the present invention are administered / used in hepatitis B patients, a significant synergistic effect is observed in the therapeutic index of chronic hepatitis B, compared to patients who have received standard treatment including the administration of conventional antiviral drugs, indicating the possibility of completely curing the disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] Figure 1a is a graph showing the changes in HBsAg levels (3-year results) in the TDF Phase 3 clinical trial of HBsAg(-) patients. TDF-TDF: TDF trial group, taking TDF in the follow-up study; ADV-TDF: ADF trial group, taking TDF in the follow-up study.

[0064] Figure 1b is a graph showing the changes in HBsAg levels (3-year results) in the TDF Phase 3 clinical trial of HBsAg(+) patients. TDF-TDF: TDF trial group, taking TDF in the follow-up study; ADV-TDF: ADF trial group, taking TDF in the follow-up study.

[0065] Figure 2 is a graph showing the importance of T cell activation in chronic hepatitis B patients.

[0066] Figure 3 This is a graph analyzing the 1 / 2a efficacy results of CVI-HBV-002.

[0067] Figure 4 This is a graph showing the results of comparison of HBeAg loss rates in the 1 / 2a clinical trial of CVI-HBV-002 and the TAF Phase 3 clinical trial.

[0068] Figure 5 is a graph demonstrating maintenance of T cell activation after administration of CVI-HBV-002.

[0069] Figure 6 It is a graph showing the results of comparison of HBeAg loss rate in the CVI-HBV-002 long-term follow-up study and the TAF Phase 3 clinical trial. DETAILED DESCRIPTION OF THE INVENTION

[0071] Hereinafter, the present invention will be described in detail.

[0072] The embodiments of the present invention may be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below. It should be understood by those skilled in the art that the embodiments of the present invention are given to explain the present invention more accurately.

[0073] In addition, throughout the specification, “comprising” one element does not exclude other elements, but may include other elements unless specifically stated otherwise.

[0074] In one aspect of the present invention, the present invention provides a pharmaceutical combination preparation for preventing or treating chronic hepatitis B, which comprises an antiviral drug for hepatitis B; and a vaccine composition comprising an antigen, a lipopeptide and a poly (I: C) adjuvant.

[0075] At this time, the antiviral drugs used for hepatitis B can be oral antiviral drugs. As specific examples of oral antiviral drugs, entecavir, tenofovir disoproxil fumarate (TDF), tenofovir rapenamide fumarate (TAF), besifovir dipivoxil maleate, lamivudine, telbivudine, rolevudine, adefovir dipivoxil, etc. can be used alone or in combination of two or more without restriction.

[0076] Below, the development trends of new chronic hepatitis B treatments are described.

[0077] Since it is difficult to cure (functionally cure) chronic hepatitis B with existing oral antiviral drugs, new chronic hepatitis B treatment methods are being actively developed. Recently, direct-acting antiviral drugs with new mechanisms have been developed, and representative types include RNA interfering agents that inhibit protein synthesis by binding to viral mRNA and breaking it down into small units or capsid inhibitors that prevent the normal production of capsids covering genetic material. On the other hand, immunotherapeutic agents with strategies to reduce HBsAg by activating the patient's weakened immune cells are being developed, and representative types include cytokines, tol-like receptor ligands that activate innate immunity, and therapeutic vaccines that control adaptive immunity by administering antigens. The development trend and types of therapeutic agents for chronic hepatitis B are shown in Table 7 in the specification of the present invention.

[0078] Regarding the current status of clinical trials of newly developed treatments for chronic hepatitis B, almost all clinical trials are being conducted with a design that confirms the therapeutic effect of combined administration of clinical trial drugs while continuing to take antiviral drugs as standard treatment. This is to protect patients by combining with standard treatment, and because it has been confirmed through clinical trials of oral antiviral drugs or combined administration studies of oral antiviral drugs and interferon that it is difficult to completely cure chronic hepatitis B with only one type of treatment. The development stages of treatments for chronic hepatitis B and whether or not antiviral drugs are administered in combination are summarized in Table 7 below.

[0079] [Table 7] Stages of development of chronic hepatitis B treatment and whether antiviral drugs are used in combination

[0080]

[0081]

[0082]

[0083] On the other hand, since the combined preparation is used for the prevention or treatment of chronic hepatitis B, the antigen is preferably the whole surface antigen (L-HBsAg) of HBV (hepatitis B virus).

[0084] In addition, an adjuvant is a substance or combination of substances that increases or induces an immune response to a vaccine antigen in a desired direction so as to enhance the clinical effect of a vaccine when used with a vaccine antigen. The main function of an adjuvant is to enhance and improve the clinical effectiveness of a vaccine, such as by acting on direct or indirect immunostimulation and antigen delivery to increase and regulate the immune response to the vaccine antigen or prolong the duration of the protective effect. When infected with pathogenic bacteria or viruses, the surface receptors (pattern recognition receptors) of immune cells recognize the unique patterns (pathogen-associated molecular patterns, PAMPs) of pathogenic microorganisms and induce an innate immune response. Toll-like receptors (TLRs) are representative surface receptors, of which about 13 types are known in humans. TLR ligands that respond to toll-like receptors are being developed as adjuvants because they directly stimulate immune cells to activate innate immune responses and induce humoral and cellular immune responses, i.e., acquired immunity against vaccine antigens to protect the human body from infectious agents or to help tissue recovery.

[0085] Lipopeptides, a specific example of adjuvants, were first synthesized by J. Metzger et al. as synthetic analogs of lipopeptides derived from bacteria and mycoplasmas (Metzger, J. et al., 1991, Synthesis of novel immunologically active tripalmitoyl-S-glycerylcysteinyllipopeptides as useful intermediates for immunogen preparations. Int. J. Peptide Protein Res. 37: 46-57). The molecular structure of the compound represented by the following formula (1) is N-palmitoyl-S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-[R]-cysteine-SKKKK (pam3Cys-SKKKK), and various other analogs have been synthesized.

[0086] [Formula (1)]

[0087]

[0088] According to H.Schild et al., when Pam3Cys-Ser-Ser is bound to an influenza virus T cell epitope and administered to mice, virus-specific cytotoxic T lymphocytes (CTLs) are induced. Generally, lipopeptides are known as ligands for TLR2. The use of such lipopeptides is not limited to Pam3Cys-SKKKK, and the lipopeptide can be composed of fatty acids and several amino acids bound to a glycerol molecule. Specific examples thereof include PHC-SKKKK, Ole2PamCys-SKKKK, Pam2Cys-SKKKK, PamCys (Pam) -SKKKK, Ole2Cys-SKKKK, Myr2Cys-SKKKK, PamDhc-SKKKK, PamCskKKK, Dhc-SKKKK, etc. The number of fatty acids in the molecule can be one or more. The number of amino acids in the lipopeptide can be one or more. In addition, fatty acids and amino acids can be chemically modified. Furthermore, the lipopeptide may be a lipoprotein, as a part of a molecule or as a whole molecule, derived from a Gram-positive or Gram-negative bacterium or a mycoplasma.

[0089] In addition, poly (I: C) has been used as an effective inducer of type 1 interferon in in vitro and in vivo studies. In addition, it is known that poly (I: C) stably and maturely forms dendritic cells, which are the most effective antigen presenting cells in mammals (Rous, R. et al 2004. poly (I: C) used for human dendritic cell maturation preserves their ability to secondarily secrete bioactive IL-12, International Immunol. 16: 767-773). According to these previous reports, poly (I: C) is an effective inducer of IL-12, which is an important cytokine that induces cellular immune responses and the formation of IgG2a or IgG2b antibodies by driving immune responses to produce Th1. In addition, poly (I: C) is known to have strong adjuvant activity against peptide antigens (Cui, Z. and F. Qui. 2005. Synthetic double stranded RNA poly I: C as a potent peptide vaccine adjuvant: Therapeutic activity against human cervical cancer in a rodent model. Cancer Immunol. Immunotherapy 16: 1-13). The length of poly (I: C) may be in the range of 50 to 5,000 bp, preferably 50 to 2,000 bp, more preferably 100 to 500 bp, but is not always limited thereto.

[0090] The lipopeptide and poly (I: C) may be included in the vaccine composition at a weight ratio of 0.1 to 10: 1, at a weight ratio of 1.25 to 2: 1, at a weight ratio of 1.25 to 1.5: 1, or at a weight ratio of 1.25: 1, but are not always limited thereto. However, the ratio may be adjusted to an appropriate level according to the patient's condition. In addition, the vaccine composition may be an aqueous solution preparation.

[0091] The vaccine composition may further include at least one selected from a pharmaceutically acceptable carrier, a diluent and an adjuvant. For example, the vaccine composition may include a pharmaceutically acceptable carrier and may be formulated for human or veterinary use and administered by various routes. The vaccine composition may be administered orally, intraperitoneally, intravenously, intramuscularly, subcutaneously and intradermally. Preferably, it is formulated into an injection and administered as an injection. Injection can be prepared using aqueous solvents such as physiological saline and Ringer's solution, vegetable oils, higher fatty acid esters (such as ethyl oleate, etc.) and non-aqueous solvents such as alcohols (such as ethanol, benzyl alcohol, propylene glycol, glycerol, etc.), and can include pharmaceutical carriers, such as stabilizers to prevent deterioration (such as ascorbic acid, sodium sulfite, sodium pyrosulfate, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for pH control, preservatives for preventing microbial development (such as chimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.) and preservatives for inhibiting microbial growth (such as phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.). Vaccine compositions can be administered in a pharmaceutically effective amount.

[0092] At this time, the term "pharmaceutically effective amount" refers to an amount sufficient to show the vaccine effect but not causing side effects or severe or excessive immune responses. The exact dosage concentration depends on the antigen to be administered and can be easily determined by a person skilled in the art based on factors well known in the medical field, such as the patient's age, weight, health condition, sex, sensitivity to drugs, route of administration and method of administration. The composition of the present invention can be administered once or multiple times. In addition, adjuvants can be further included in the vaccine composition as other known adjuvants in addition to lipopeptides and poly (I: C).

[0093] In another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating chronic hepatitis B, which comprises an antiviral drug for hepatitis B; and a vaccine composition comprising an antigen, a lipopeptide and a poly (I:C) adjuvant.

[0094] In another aspect of the present invention, the present invention provides a method for producing an immune response against chronic hepatitis B in a subject, the method comprising the step of administering to the subject a drug combination preparation for preventing or treating chronic hepatitis B, the drug combination preparation comprising an antiviral drug for hepatitis B; and a vaccine composition comprising an antigen, a lipopeptide and a poly (I:C) adjuvant.

[0095] If the vaccine composition is administered to humans (patients), it can be administered in an amount effective to stimulate immune response in vivo, for example, it can be administered to humans once or several times at a dose of 1-250 μg, more preferably 10-100 μg, but is not always limited thereto.

[0096] In another aspect of the present invention, the present invention provides a combination preparation kit for preventing or treating chronic hepatitis B, which comprises a first component containing an antiviral drug for hepatitis B; and a second component containing a vaccine composition, which vaccine composition includes an antigen, a lipopeptide and a poly (I: C) adjuvant.

[0097] In another aspect of the present invention, the present invention provides a method for preventing, improving or treating chronic hepatitis B, which comprises the steps of administering to a subject an antiviral drug for hepatitis B; and a vaccine composition containing an antigen, a lipopeptide and a poly (I:C) adjuvant.

[0098] In another aspect of the present invention, the present invention provides an antiviral drug for hepatitis B; and use of a vaccine composition comprising an antigen, a lipopeptide and a poly (I:C) adjuvant in the preparation of a drug for preventing, improving or treating chronic hepatitis B.

[0099] When the pharmaceutical composition, pharmaceutical combination preparation and combination preparation kit provided by one aspect of the present invention are administered / used in patients with hepatitis B, a significant synergistic effect is observed in the therapeutic index of chronic hepatitis B compared to patients who have undergone standard treatment including administration of conventional antiviral drugs, indicating the possibility of complete cure of the disease. This is directly supported by the clinical trials of the following embodiments and experimental examples.

[0100] Hereinafter, the present invention will be described in detail through the following Preparation Examples, Examples and Experimental Examples.

[0101] However, the following preparation examples, examples and experimental examples are only used to illustrate the present invention, and the contents of the present invention are not limited thereto.

[0102] Preparation of a vaccine composition containing antigen, lipopeptide and poly (I:C) adjuvant (CVI-HBV-002 therapeutic vaccine)

[0103] Recently, there has been a growing demand for the development of immunotherapeutic agents that can lead to a reduction in the virus in the body by addressing the reduction in immune tolerance in patients with chronic hepatitis B. The therapeutic vaccine CVI-HBV-002 consists of a third-generation antigen L-HBsAg with excellent immunogenicity and a potent adjuvant L-pampo [Pam3Cys-SKKKK+poly(I:C)]. When it is administered in combination with antiviral drugs, a synergistic effect of inhibiting hepatitis B virus proliferation, addressing the patient's immune tolerance, and activating T cells can be expected. The ultimate goal of developing CVI-HBV-002 is to induce adaptive immunity to hepatitis B virus, thereby achieving rapid cure (see Figure 2More specifically, 200 or 400 μg of poly(I:C) was added to 20 or 40 μg of total surface antigen (L-HBsAg), mixed thoroughly, and then 250 or 500 μg of Pam3Cys-SKKKK (a lipopeptide) was added to the mixture to prepare the therapeutic vaccine CVI-HBV-002.

[0104] Combination therapy for hepatitis B patients receiving standard therapy

[0105] A clinical trial was conducted in which oral antiviral drugs and the therapeutic vaccine CVI-HBV-002 prepared above were co-administered to chronic hepatitis B patients who had already received standard treatment including the administration of antiviral drugs. In each experimental example described below, specific details such as the number of patients actually recruited, gender, age, and the type and dose of antiviral drugs that had been prescribed are described. On the other hand, unless otherwise stated, in the experimental examples described below, chronic hepatitis B patients took antiviral drugs orally once a day, and the therapeutic vaccine CVI-HBV-002 prepared above was administered by intramuscular injection three or six times every four weeks or the therapeutic vaccine CVI-HBV-002 prepared above was administered six times every two weeks together with the oral antiviral drug from the start of the combined administration.

[0106] 1 / 2a Clinical Trial Results

[0107] 1-1.1 / 2a Clinical trial efficacy results

[0108] The safety and tolerability of the therapeutic vaccine CVI-HBV-002 for hepatitis B were confirmed by a Phase 1 / 2a clinical trial in which oral antiviral drugs and the therapeutic vaccine CVI-HBV-002 prepared as described above were co-administered to patients with chronic hepatitis B who had already received standard treatment including the administration of conventional antiviral drugs.

[0109] A total of 53 patients were recruited for the 1 / 2a clinical trial, of which 51 patients were included in the efficacy evaluation analysis except for 2 patients who withdrew. Among them, 35 were male (68.6%) and 16 were female (31.4%), with an average age of 46 years old. The average length of time the subjects took oral antiviral drugs was about 70 months. Regarding the type of oral antiviral drugs, 21 patients took TDF alone, 14 patients took entecavir alone, and 11 patients took adefovir and other types of oral antiviral drugs (entecavir, lamivudine or telbivudine) at the same time. In addition, there was 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 drugs (clovudine, adefovir and TDF) at the same time. The drug is administered orally once daily, with the dose determined by each antiviral drug (equivalent: entecavir 0.5 mg, TDF 300 mg, adefovir 10 mg).

[0110] The antigen dose of the hepatitis B therapeutic vaccine CVI-HBV-002 was 20 μg or 40 μg, 3 times every 4 weeks, 6 times every 4 weeks, or 6 times every 2 weeks, depending on the grouping of the subjects.

[0111] The results are shown in Figure 3 middle.

[0112] Figure 3 This is a graph showing the results of analyzing the 1 / 2a effectiveness of CVI-HBV-002.

[0113] like Figure 3 As shown, it was confirmed that HBV-specific T-cell immune responses were induced in 41 of 48 patients (85.4%) after vaccination, resulting in overcoming immune tolerance by vaccination. In addition, a very encouraging result was obtained, in which 8 of 35 subjects (23%) who remained HBeAg-positive after long-term use of antiviral drugs induced HBeAg serum loss, and 36 of 51 patients (70.6%) showed a quantitative reduction in serum HBsAg. Figure 3 The above results are highlighted in red. From these results, it can be confirmed that HBV-specific T cells were activated by the administration of CVI-HBV-002, which greatly affected the quantitative reduction of HBsAg and the loss of HBeAg.

[0114] 1-2. Comparison of the results of Gilead's therapeutic vaccine (GS4774) and clinical trial controls

[0115] To make up for the lack of a control group that only took antiviral drugs in the Phase 1 / 2a clinical trial, the control group results of Gilead's therapeutic vaccine (GS4774) Phase 2 clinical trial were compared (Randomized phase II study of GS-4774 as a therapeutic vaccine in virally suppressed patients with chronic hepatitis B (Journal of Hepatology 2016 vol.65 509-516)).

[0116] The results are shown in Tables 8 and 9 below.

[0117] [Table 8] Comparison of clinical trial results with competing products

[0118]

[0119]

[0120] As shown in Table 8, Gilead's Phase 2 clinical trial failed because it did not show a significant difference from the control group in the HBsAg change rate, which was the primary endpoint. The HBeAg loss rate in the GS4774-administered group was only 11%, which was significantly different from the 23% HBeAg loss rate in the CVI-HBV-002 1 / 2a clinical trial.

[0121] [Table 9] Comparison with the results of the control group in the Phase 2 clinical trial of GS4774

[0122]

[0123]

[0124] As shown in Table 9, as a result of comparing the results of the control group of the Phase 2 clinical trial using GS4774 with the results of the CVI-HBV-002 clinical trial, the control group of the Phase 2 clinical trial of GS4774 showed no HBeAg loss in the 1-year evaluation, which was significantly different from the 23% HBeAg loss rate in the 1 / 2a clinical trial of CVI-HBV-002. In addition, in the case of HBsAg reduction, the 1 / 2a results analyzed at the same evaluation time point as the efficacy evaluation time point of the GS4774 clinical trial were used for comparison. The results confirmed that the reduction in HBsAg in the 1 / 2a clinical trial was greater than that in the control group of the Phase 2 clinical trial of GS4774.

[0125] Comparison with the clinical trial results of GS4774 confirmed that it is difficult to overcome the immune tolerance of patients with chronic hepatitis B with oral antiviral drugs alone, and co-administration of CVI-HBV-002 is more effective in HBeAg loss or HBeAg reduction.

[0126] 1-3. Comparison of TAF Phase 3 Clinical Trial Results (1 Year)

[0127] Secondly, the results of the Phase 3 clinical trial (1 year) of TAF were compared with the results of the 1 / 2a clinical trial of CVI-HBV-002. In the Phase 3 clinical trial of TAF, TDF was used as an active control group, and the detailed results of this clinical trial were the same as the results of the Phase 3 clinical trial of tenofovir alafenamide (TAF) described in the description of the above-mentioned related art [Table 4]. The results of the TAF Phase 3 clinical trial (including the active control TDF results) were used to compare the 1 / 2a efficacy results of CVI-HBV-002 because the subjects in the oral antiviral drug clinical trial were most similar to those in the 1 / 2a clinical trial of CVI-HBV-002, including patients with existing antiviral drug treatment experience in the TAF Phase 3 clinical trial. In the Phase 3 clinical trial of TAF, the proportion of subjects with antiviral treatment experience was approximately 20% to 26%, although it varied slightly by group.

[0128] The results are as follows Figure 4 As shown in Table 10.

[0129] Figure 4 This is a graph showing the results of comparison of HBeAg loss rates in the CVI-HBV-002 1 / 2a clinical trial and the TAF Phase 3 clinical trial.

[0130] [Table 10] Comparison of CVI-HBV-002 Phase 1 / 2a and TAF Phase 3 clinical trial results (1 year)

[0131]

[0132]

[0133] like Figure 4As shown in Table 10, when comparing the efficacy, CVI-HBV-002 showed the highest HBeAg loss rate of 23%, followed by TAF at 14% and TDF at 12%. Considering that the efficacy evaluation time points of the Phase 1 / 2a clinical trial were significantly earlier than the TAF clinical trial evaluation time points (Week 48, 1 year) by 20 weeks, 22 weeks and 32 weeks compared with the baseline time points depending on the group, the above results are very encouraging and show the synergistic effect of the combined administration of CVI-HBV-002. In addition, no subject in the CVI-HBV-002 group showed HBsAg loss, the HBsAg loss rate in the TAF group was less than 1%, and only 1 of 428 patients in the TDF group showed HBsAg loss. At the same time, HBV-specific T cell responses are not included in the evaluation items of clinical trials of oral antiviral drugs, so they cannot be compared.

[0134] Results of the Phase 1 / 2a Long-term Follow-up Study of CVI-HBV-002

[0135] 2-1. Efficacy results of the long-term follow-up study of the 1 / 2A clinical trial

[0136] A long-term follow-up study was conducted to confirm the efficacy of the hepatitis B therapeutic vaccine (CVI-HBV-002) in subjects participating in the Phase 1 / 2a clinical trial, and a total of 24 patients participated in the study.

[0137] The results are shown in Table 11.

[0138] [Table 11] Summary of long-term follow-up efficacy evaluation results

[0139]

[0140]

[0141] As shown in Table 11, the results of quantitative analysis of HBsAg confirmed that HBsAg levels were further reduced in 15 patients, of which 5 patients showed HBsAg reduction rates of 50% or more and 2 patients showed HBsAg reduction rates of 99% or more. The results of analyzing HBeAg serum loss showed that two more patients showed HBeAg serum loss compared with the results 12 weeks after the last CVI-HBV-002 administration in the 1 / 2a clinical trial.

[0142] Next, when HBV-specific T cell immune responses were analyzed, 8 of 14 subjects (57.1%) showed increased formation of IFN-γ spots compared with the 1 / 2a clinical trial baseline, confirming that T cell activation was sustained (see Figure 5). Among them, 6 patients (42.9%) showed the formation of IFN-γ spots more than twice the baseline. Through these results, it was confirmed that the immune activation of the therapeutic vaccine CVI-HBV-002 lasted for a long time, once again confirming the possibility of treating chronic hepatitis B.

[0143] 2-2. Comparison of the long-term follow-up results of CVI-HBV-002 and the results of the TAF Phase 3 clinical trial (2 years)

[0144] The results of the long-term follow-up study of subjects enrolled in the Phase 1 / 2a clinical trial approximately 4 years later were compared with the results of the second-year efficacy evaluation of the TAF Phase 3 clinical trial. Figure 6 As shown in Table 12.

[0145] Figure 6 It is a graph showing the results of comparison of HBeAg loss rate in the CVI-HBV-002 long-term follow-up study and the TAF Phase 3 clinical trial.

[0146] [Table 12] Comparison of long-term follow-up results of CVI-HBV-002 and TAF Phase 3 clinical trial results (2 years)

[0147]

[0148]

[0149] *Subjects with very high HBV DNA and HBsAg levels were excluded due to low compliance with antiviral medications.

[0150] like Figure 6 As shown in Table 12, the results of the long-term follow-up study conducted on subjects enrolled in the Phase 1 / 2a clinical trial an average of 4 years later were compared with the second-year efficacy evaluation results of the Phase 3 clinical trial of TAF. The HBeAg loss rate after administration of CVI-HBV-002 was still the highest.

[0151] Regarding the HBeAg loss rate, the HBeAg loss rate was 30% in the CVI-HBV-002 group, 22% in the TAF group, and 18% in the TDF group.

[0152] In order to make up for the lack of a control group in the 1 / 2a clinical trial using CVI-HBV-002, other clinical trials that could be replaced by possible control groups were studied and the efficacy was compared. The results confirmed that the combined administration of CVI-HBV-002 was more effective than the administration of antiviral drugs alone. In the clinical trial of Gilead's therapeutic vaccine (GS4774), no patients in the group that only administered antiviral drugs showed HBeAg loss, and the degree of T cell activation was lower than that in the group administered GS4774, while after CVI-HBV-002 administration, 23% of patients showed HBeAg loss and 85.4% of patients showed T cell activation. These results confirm that CVI-HBV-002 overcomes the immune tolerance of patients with chronic hepatitis B.

[0153] In addition, compared with the efficacy results of the Phase 3 clinical trial of TAF, which also included patients taking antiviral drugs, although the efficacy evaluation time point after CVI-HBV-002 administration was earlier, the HBeAg loss rate was the highest (23%). From the results, it can be confirmed that the combined administration of CVI-HBV-002 has a higher therapeutic effect than TAF (14%) or TDF (12%) alone. When comparing the results of 2-year administration of oral antiviral drugs and the long-term follow-up results of CVI-HBV-002, CVI-HBV-002 had the highest HBeAg loss rate of 30%. The 1 / 2a clinical trial using CVI-HBV-002 was initially conducted on a small scale, which made it difficult to confirm HBsAg loss, but since HBeAg loss is preceded by HBsAg loss, a higher HBeAg loss rate than oral antiviral drugs is an encouraging result. These results support the possibility of combination therapy, which achieves rapid cure by administering CVI-HBV-002 and antiviral drugs together so that the antiviral drugs can have a synergistic effect on the inhibition of HBV proliferation.

Claims

1. A pharmaceutical combination preparation for preventing or treating chronic hepatitis B, comprising an antiviral drug for hepatitis B; and a vaccine composition containing an antigen, a lipopeptide and a poly (I: C) adjuvant, wherein the antiviral drug for hepatitis B is an oral antiviral drug, wherein the oral antiviral drug is selected from at least one of entecavir, tenofovir disoproxil fumarate (TDF), tenofovir alafenamide fumarate (TAF), besifovir dipivoxil maleate, lamivudine, telbivudine, clovudine and adefovir dipivoxil, and the The antigen is the whole surface antigen (L-HBsAg) of HBV (hepatitis B virus), and wherein the lipopeptide is at least one selected from Pam3Cys-SKKKK, PHC-SKKKK, Ole2PamCys-SKKKK, Pam2Cys-SKKKK, PamCys(Pam)-SKKKK, Ole2Cys-SKKKK, Myr2Cys-SKKKK, PamDhc-SKKKK, PamCSKKK and Dhc-SKKKK.

2. The pharmaceutical combination preparation for preventing or treating chronic hepatitis B according to claim 1, wherein the lipopeptide and the poly (I:C) are contained in a weight ratio of 0.1 to 10:

1.

3. The pharmaceutical combination preparation for preventing or treating chronic hepatitis B according to claim 1, wherein the vaccine composition is an aqueous solution preparation.

4. The pharmaceutical combination preparation for preventing or treating chronic hepatitis B according to claim 1, wherein the vaccine composition further comprises at least one selected from a pharmaceutically acceptable carrier, a diluent and an adjuvant.

5. The pharmaceutical combination preparation for preventing or treating chronic hepatitis B according to claim 1, wherein the vaccine composition is administered by any one of the following administration routes: oral, transdermal, intramuscular, intraperitoneal, intravenous, subcutaneous and nasal administration.

6. A combination preparation kit for preventing or treating chronic hepatitis B, comprising a first component containing a hepatitis B antiviral drug; and a second component containing a vaccine composition, the vaccine composition comprising an antigen, a lipopeptide and a poly (I: C) adjuvant, wherein the antiviral drug for hepatitis B is an oral antiviral drug, wherein the oral antiviral drug is selected from entecavir, tenofovir disoproxil fumarate (TDF), tenofovir alafenamide fumarate (TAF), besifovir dipivoxil maleate, lamivudine, telbivudine, clovudine and adefovir dipivoxil At least one of the above, the antigen is the whole surface antigen (L-HBsAg) of HBV (hepatitis B virus), and wherein the lipopeptide is at least one selected from Pam3Cys-SKKKK, PHC-SKKKK, Ole2PamCys-SKKKK, Pam2Cys-SKKKK, PamCys(Pam)-SKKKK, Ole2Cys-SKKKK, Myr2Cys-SKKKK, PamDhc-SKKKK, PamCSKKK and Dhc-SKKKK.

7. Use of the pharmaceutical combination preparation according to any one of claims 1 to 5 in the preparation of a medicament for preventing or treating chronic hepatitis B.

8. Use of the pharmaceutical combination preparation according to any one of claims 1 to 5 in the preparation of a medicament for improving chronic hepatitis B.

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