TW201632192A - A kit for influenza vaccine and use thereof - Google Patents

Abstract

The present invention relates to a kit for influenza vaccine, comprising: a first vaccine composition including a Flu A of H1N1, a Flu A of H3N2, and a first Flu B; and a second vaccine composition including a second Flu B different from the first Flu B. A use method of the kit is also provided herein, wherein the kit is administrated to an individual. The method comprises the following steps: a. administrating a first vaccine composition to the individual; and b. administrating a second vaccine composition to the individual after 1 to 10 days of the administration of the first vaccine composition. Thereby, the prevention from Influenza will be more effective and broader.

Description

Influenza vaccine kit and method of use thereof

The present invention relates to an influenza vaccine kit and method of use thereof, and more particularly to a second vaccine composition comprising: a first vaccine composition comprising a trivalent influenza vaccine and a monovalent influenza vaccine.

According to the influenza virus as the main human pathogen, it causes respiratory diseases (commonly known as "flu" or "influenza"), and the severity can range from sub-clinical infection to the primary cause of death. Within the scope of viral pneumonia. It is estimated that approximately 1 billion people worldwide suffer from influenza-infected diseases each year, causing serious illnesses in 3 to 5 million cases and causing an estimated 300,000 to 500,000 influenza-related deaths.

Most of these infectious diseases can be attributed to influenza A viruses carrying H1 or H3 hemagglutinin subtypes, a small part of which is attributed to influenza B virus, so representatives of these three types of influenza viruses are included in seasonal In the vaccine.

Influenza A viruses are widely distributed in nature and can infect a variety of birds and mammals. The influenza virus is an enveloped RNA virus belonging to the family of the Orthomyxoviridae family. Although all known influenza A virus subtypes are found in poultry, the currently circulating human influenza A subtypes are H1N1 and H3N2. Influenza B strains are strictly directed against humans.

The antigenic variation of HA in influenza B virus strain is less than that in type A virus The antigenic variation observed in the strain. Two strains of influenza B virus: Victoria lineage (Virus line B, Victorias/2/87) and Yamagata lineage (B/Yamagata/16/88) have been popular worldwide since the late 1980s. The health authorities are known to be prevalent every year for the four major influenza strains, causing most of the flu diseases, but the flu vaccine used in the past three decades can only fight against three strains. Influenza B viruses become the main seasonal infectious disease every 2-4 years. The above two types of influenza B viruses use the Hemagglutination-inhibiting (HAI) test, which is mostly lacking antigen cross-reaction and cannot be cross-protected.

Current immunization practices rely on early identification of a circulating influenza virus to allow timely production of an effective seasonal influenza vaccine. Because the two influenza B viruses are not cross-protective, global influenza experts have to decide which strain to include as a seasonal viral flu vaccine among the strains covered in each season, but this decision is difficult and difficult. Precisely, in the flu season of the past 11 years (2001-2012), there are six years, and the main strains that are prevalent in the season do not match the seasonal flu vaccine. There are inherent difficulties in predicting strains that will dominate during the next season.

In order to deal with the inability of this type B influenza virus vaccine to match the major strains that are prevalent in the season, vaccine manufacturers have begun to prepare "tetravalent" vaccines containing the following four influenza strains: two influenza A strains. And two strains of influenza B virus. This four-valent vaccine can provide a wider range of protection. For example, there is currently an intranasal tetravalent influenza vaccine (FluMist Quadrivalent MedImmune, LLC) and three injectable influenza vaccines (including: Fluarix Quadrivalent GlaxoSmithKline Biologicals, Fluzone Quadrivalent Sanofi Pasteur Inc. And FluLaval Quadrivalent ID Biomedical Corporation.) was approved by the US authorities.

The seasonal influenza vaccine must be updated annually because of the variability of the virus. Due to the constant mutational evolution of influenza viruses, there are different new strains of influenza each year, so there is still a need for safe and effective influenza vaccines to stimulate a robust and broadly neutralizing antibody response, and widely targeted at current And future influenza strains (seasonal and pandemic) provide protection for the effective prevention and treatment of influenza. .

On the other hand, the applicant's previous study found that after the injection of the tetravalent vaccine, the additional influenza B virus may be inhibited by H1N1 and H3N2 in the influenza A virus in the tetravalent vaccine, thus producing a specific rate of influenza B virus. The vaccine has a lower HAI titer. Therefore, in order to solve the problem that the current tetravalent vaccine is difficult to induce sufficient Flu B antiviral HAI titer, it is necessary to develop a more effective tetravalent vaccine to induce high antibody titer to meet the protection criterion of HAI titer ≧40.

In view of the deficiencies of the prior art, it is an object of the present invention to provide an influenza vaccine kit and a method of using the same that provides a more effective and comprehensive protection than conventional influenza vaccines.

Another object of the present invention is to provide an influenza vaccine kit and a method for using the same, which can have higher antiviral HAI titer against influenza virus B, and its antiviral HAI titer against influenza A virus is also maintained at a stable rate. degree.

The present invention provides an influenza vaccine kit comprising: a first vaccine composition comprising: an H1N1 influenza A virus, an H3N2 influenza A virus, and a first influenza B virus; A second vaccine composition comprising: a second influenza B virus different from the first influenza B virus.

In one embodiment, the H1N1 influenza A virus may be: A/California/7/2009 virus or A/California/7/2009 (H1N1) pdm09 virus (A/California/7/ 2009 (H1N1) pdm09-like virus).

In one embodiment, the H3N2 type A influenza virus may be: A/Texas/50/2012 type virus or virus antigenic similar to the cell proliferation prototype A/Victoria/361/2011 type virus.

In one embodiment, the first influenza B virus or the second influenza B virus may be a Victoria lineage or a Yamagata lineage influenza B virus.

In one embodiment, the first influenza B virus can be: a B/Brisbane/60/2008 virus or a B/Massachusets/2/2012 virus.

In another embodiment, the second influenza B virus can be: a B/Brisbane/60/2008 virus or a B/Massachusets/2/2012 virus.

In one embodiment, the H1N1 influenza A virus, the H3N2 influenza A virus, the Victoria lineage influenza B virus, and the Yamagata lineage influenza B virus The dose can be from 1 to 30 μg, respectively.

In one embodiment, the influenza vaccine kit may further comprise a pharmaceutically acceptable carrier, which may comprise one or more groups selected from the group consisting of: a solvent, an emulsifier ( Emulsifier), suspending agent, decomposer, binding agent, Excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, lubricant, surfactant Surfactant, adjuvant, and other carriers similar or suitable for use in the present invention.

In one embodiment, the influenza vaccine kit can be prepared as a group consisting of one or more of the following dosage forms: an injection form, an oral form, a surface administration type, and an inhalation type.

The present invention also provides a method of using the influenza vaccine kit described above, wherein the influenza vaccine kit is applied to a body and comprises the steps of: a. administering a first vaccine composition to the individual; and b. administering A second vaccine composition is administered to the individual about 1 to 10 days after the first vaccine composition.

In one embodiment, the method of use may further comprise the step of administering a tetravalent influenza vaccine to the individual prior to the step a. For example, in one embodiment, the tetravalent influenza vaccine can be pre-administered to the individual about at least two weeks prior to the step a.

In one embodiment, the tetravalent influenza vaccine may comprise: an H1N1 influenza A virus, an H3N2 influenza A virus, a Victoria lineage influenza B virus, and a Yamagata type ( Yamagata lineage) type B flu virus.

In one embodiment, the H1N1 influenza A virus, the H3N2 influenza A virus, the Victoria lineage influenza B virus, and the Yamagata lineage in the tetravalent influenza vaccine The dose of influenza B virus can be 1 to 30 μg, respectively.

In one embodiment, the H1N1 influenza A virus may be: A/California/7/2009 virus or A/California/7/2009 (H1N1) pdm09 virus (A/California/7/ 2009 (H1N1) pdm09-like virus).

In one embodiment, the H3N2 type A influenza virus may be: A/Texas/50/2012 type virus or virus antigenic similar to the cell proliferation prototype A/Victoria/361/2011 type virus.

In one embodiment, the Victorian lineage influenza B virus can be a B/Brisbane/60/2008 virus.

In one embodiment, the Yamagata lineage influenza B virus can be a B/Massachusets/2/2012 virus.

First Fig. In the test example of the present invention, the vaccine of the experimental group and the control group elicited an immune response in the mouse. Among them, the mice were vaccinated with a tetravalent vaccine of the control group or a vaccine set of the experimental group (the first vaccine composition containing the trivalent vaccine + the second vaccine composition containing the monovalent influenza virus B). Mouse antiserum was collected to assess HAI titers for the following different influenza viruses: Figure (A) H1N1 (A/California/7/2009), Figure (B) H3N2 (A/Texas/50/2012), Figure (C) Flu B/Brisbane/60/2008 and (D) Flu B/Massachusetts/02/2012.

Next, embodiments of the present invention are described in detail by the following examples, but are not limited thereto. The above and other objects, features and advantages of the present invention will become more apparent from

In the present invention, the following noun definitions are used, but it should be understood that This definition is only used to help explain and explain the contents of the present invention, and is not intended to limit the scope of the present invention.

The term "influenza vaccine" as used in the present invention refers to any preparation intended to be administered to a subject to produce or artificially increase the immune response to a particular disease (eg, any disease caused by the influenza virus in the present invention) in the subject. . The term "trivalent" or higher as referred to herein means the immunity against three (or more) vaccines against three (or more) viruses (three or more) for three (or more) diseases or serotypes. . These trivalent (or higher) vaccines are prepared from several different types of antigens.

The term "....." used in the present invention (for example, A/California/7/2009 virus, B/Brisbane/60/2008 virus or B/Massachusets/2/2012) Viral) refers to a strain or isolate of influenza virus, which can encompass natural isolates as well as artificial mutants or recombinant strains and analogs thereof, since they are usually produced by mutations and exhibit different pathogenic profiles. Thus, as used herein, the terms "viral strain" and "isolated strain" are used interchangeably. The current nomenclature for human influenza strains or isolates includes the type of virus (genus) (ie, A, B, or C), the geographic location of the first isolation, the number of strains, and the year of isolation, usually given in brackets for HA and NA. Description of the antigen, for example: A/California/7/2009 (H1N1) pdm09. Non-human strains also include the originating host in the nomenclature.

The term "individual" as used in the present invention includes animals, generally mammalian species, such as humans or rodents, and domesticated animals such as dogs, cats, horses, and the like.

The term "pharmaceutically acceptable carrier" as used in the present invention may contain inert ingredients which do not excessively inhibit the biological activity of the compound. Pharmaceutically acceptable carriers should be biocompatible, for example, non-toxic, non-inflammatory, non-immunogenic or when administered Does not produce other undesirable reactions or side effects. Standard pharmaceutical blending techniques are available. Some examples of materials that can be used as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (eg, human serum albumin), buffer substances (eg, Twin 80, phosphate, glycine, sorbic acid or potassium sorbate), a mixture of glycerides of saturated plant fatty acids, water, salt or electrolyte (eg potassium sulphate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride) Or zinc salt), colloidal vermiculite, magnesium tricaprate, polyvinylpyrrolidone, polyacrylate, wax, polyethylene-polypropylene oxide-block polymer, methylcellulose, hydroxypropylmethyl Cellulose, lanolin, sugar (such as lactose, glucose and sucrose); starch (such as corn starch and potato starch); cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate) Powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; Such as propylene glycol or polyethylene glycol; esters, such as ethyl oleate and Ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol and phosphate buffer solution, and other non-toxic phases Lubricating agents, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavoring and flavoring agents, preservatives and antioxidants may also be present in the compositions, It depends on the judgment of the staff.

The term "adjuvant" as used in the present invention refers to a substance which is added to an immunological composition such as a vaccine and which does not have any single antigenic action itself, which stimulates the immune system and increases the immune response to the immune composition, adjuvant Examples may include, but are not limited to, oily adjuvants (eg, mineral oil, vegetable oil, animal oil, complete Freund's adjuvant, Freund's incomplete adjuvant, etc.), aqueous adjuvants (eg, aluminum hydroxide), biphasic Oily adjuvants (eg water-in-water-in-water dosage form, w/o/w), bio-type adjuvants (eg CpG-oligomers) Nucleotides, bacterial toxoids, etc.). Wherein the dual phase oily adjuvant comprises a surfactant and an oil phase material; the surfactant comprises one or more of the following selected groups: sorbitol fatty acid ester; sorbitol fatty acid Ester and ethylene oxide or propylene oxide concentrate; mannitol fatty acid ester; mannitol fatty acid ester and ethylene oxide or propylene oxide concentrate; mannitol fat Acid esters with the following selected hydrophilic groups: carboxylic acid, amine, amide, alcohol, polyester, ether, oxygen Oxide conjugate; anhydrous mannitol fatty acid ester; anhydrous mannitol fatty acid ester with the following selected hydrophilic groups: carboxylic acid, amine, decylamine, alcohol, polyester polyol, ether a conjugate of an oxy group; a saccharose fatty acid ester; a sucrose fatty acid ester with an ethylene oxide or propylene oxide concentrate; a glycerin fatty acid ester; a glycerin fatty acid ester with ethylene oxide or propylene oxide Concentrate; fatty acid and ethylene oxide or propylene oxide concentrate; fatty alcohol and epoxy Dioxane or propylene oxide concentrate; and glycerophospholipids (glycerophospholipid). The oil phase material includes one or more of the following selected groups: mineral oil, vegetable oil, and animal oil.

The term "dosage form" as used in the present invention can be classified into an injection type (e.g., an injection solution), an oral type (e.g., a tablet, a capsule, a granule, a powder, an oral suspension), and a surface application type (transdermal). Patch or ointment, gel, balm, lotion or eye wash, etc.) and inhalation type. Even sublingual, buccal, intratracheal, intraocular, intranasal, atrial dosage forms, rectal dosage forms or implants may be used as appropriate. For example, the vaccine kit of the present invention may be administered by a method such as oral, sublingual, subcutaneous, intramuscular, intravenous, superficial, topical, intratracheal, intranasal, transdermal, rectal, intraocular or atrial. Different dosage forms are administered by means of internal administration or the like.

First, the preparation example

The steps of the preparation examples are carried out in the following steps: 1. Calculate the theoretical requirements of each MPH in Table 1 below; 2. Calculate the theoretical requirement of 0.01 M PBS pH 7.1; 3. Transfer the calculated theoretical requirement of 0.01 M PBS pH 7.1 to a 5 mL Vial bottle, and then calculate the theoretical requirement of each MPH into 5 mL containing 0.01 M PBS pH 7.1. The Vial bottle was mixed and filled, and after the product label was attached, the samples of the control group and the experimental group were completed. *All samples are prepared before immunization

(1) Experimental materials

The materials used in the preparation examples are shown in Table 2 below.

Table 2 Materials used in the preparation examples

(2) Experimental process:

1. Animal breed BALB/c female mice (about 7-8 weeks old, 16-18 g).

2. Prophylactic injection: On the 0th day (first injection) and the 14th day (additional injection) of the vaccination period, the mice were intramuscularly injected with a dose of 0.05 ml of the vaccine sample.

3. Blood collection: Antiserum was collected from the injected mice on the 34th day of the vaccination period.

Second, the test case

(1) The experimental group used in the test case and the vaccine composition used therein, the antiserum obtained in the above preparation example was subjected to the following hemagglutination inhibition test to measure the immunoreactivity and neutralizing titer of the vaccine-induced HA reactivity. . The mouse group in the control group and the experimental group used in the test example and the vaccine composition used therein are shown in Table 2 below.

Table 2 Mouse group and vaccine composition in the test case

(2) Hemagglutination inhibition test

The serum was double-diluted at an initial dilution ratio of 1:10 to carry out the following test. This serum was then mixed with 8 HA units of influenza virus A/California/7/2009, A/Texas/50/2012, B/Brisbane/60/2008 and B/Massachusetts/02/2012, respectively. After incubation at room temperature for 1 hour, freshly prepared 0.5% turkey red blood cells were added, and after 1 hour, hemagglutination conditions were visually evaluated. The hemagglutination inhibition (HAI) titer is defined as the reciprocal of the highest dilution factor showing greater than 50% hemagglutination inhibition. If there is no inhibition at a serum dilution ratio of 1:10, the titer is recorded as 5.

(3) Experimental results

This test case is designed to test the efficacy of the experimental group's kit. Please refer to Table 2, each group of 10 mice to contain H1N1 (A/California/7/2009), Commercially available tetravalent vaccines (provided with the brand name) of H3N2 (A/Texas/50/2012), Flu B/Brisbane/60/2008 and Flu B/Massachusetts/02/2012 for vaccination, antibiotics of each group Serum was analyzed for the (Flu B) HAI titer against H1N1, H3N2 and influenza B viruses, respectively.

The results of this test example show that the sera of the vaccination with the tetravalent vaccine (control group) yielded the following data: The titer of the H1N1 (A/California/7/2009) virus in the first panel A was: HAI ≧ 80 -160, GMT = 108.5; the titer of H3N2 (A/Texas/50/2012) virus in the first panel B is: HAI ≧ 320-640, GMT = 436; in Figure C, Flu B/Brisbane/ The titer of the 60/2008 virus was: HAI ≧ 40-80, GMT = 51.26; the titer of the Flu B/Massachusetts/02/2012 virus in the first panel D was: HAI ≧ 40-80, GMT = 78.5.

Interestingly, the experimental group of the present invention was injected with a trivalent vaccine (H1N1 (A/California/7/2009), H3N2 (A/Texas/50/2012), and after the first injection. Flu B/Brisbane/60/2008) first vaccine composition, followed by injection of a second vaccine composition of monovalent vaccine on day 1, day 3 and day 6 after additional injection (Flu B/Massachusetts/ 02/2012), it can be confirmed from the first figure that the experimental group in this case elicited a higher anti-Flu B/Brisbane/60/2008virus or Flu B/Massachusetts/02/2012 Flu compared to the control group's tetravalent vaccine. B antibody titer.

The results of the experiment can confirm that the influenza vaccine kit provided by the present invention can solve the problem that the current tetravalent vaccine is difficult to induce sufficient influenza virus B antiviral HAI titer, can induce high antibody titer, and can meet the HAI effect. The protection criteria for price ≧40. Furthermore, compared with the tetravalent vaccine of the control group, the vaccine kit of the experimental group of this case can achieve stable anti-influenza A (H1N1 and H3N2) virus titers, and Compared with the control group, the two strains of influenza virus B: Victoria line (Victoria lineage, Viruses B/Victoria/2/87) and Yamagata line (Yamagata lineage, B/Yamagata/16/88) are better. HAI.

It will be apparent to those skilled in the art that various changes can be made in accordance with the embodiments of the present invention without departing from the spirit of the invention. Therefore, it is to be understood that the invention is not limited by the scope of the invention, and is intended to cover the modifications of the spirit and scope of the invention.

Claims (18)

An influenza vaccine kit comprising: a first vaccine composition comprising: an H1N1 influenza A virus, an H3N2 influenza A virus, and a first influenza B virus; and a second vaccine A composition comprising: a second influenza B virus different from the first influenza B virus. The influenza vaccine kit according to claim 1, wherein the H1N1 influenza A virus is a virus of A/California/7/2009 or A/California/7/2009 (H1N1) pdm09. Virus (A/California/7/2009 (H1N1) pdm09-like virus). The influenza vaccine kit according to claim 1, wherein the H3N2 influenza A virus A/Texas/50/2012 virus or virus antigenicity is similar to the cell proliferation prototype A/Victoria/361/ 2011 virus. The influenza vaccine kit of claim 1, wherein the first influenza B virus or the second influenza B virus is a Victoria lineage or a Yamagata lineage influenza B virus. The influenza vaccine kit of claim 4, wherein the first influenza B virus is a B/Brisbane/60/2008 virus or a B/Massachusets/2/2012 virus. The influenza vaccine kit of claim 4, wherein the second influenza B virus is a B/Brisbane/60/2008 virus or a B/Massachusets/2/2012 virus. The influenza vaccine kit according to claim 1, wherein the H1N1 influenza A virus, the H3N2 influenza A virus, the Victoria lineage influenza B virus, and the Yamagata type (Yamagata lineage) The dose of influenza B virus is 1 to 30 μg, respectively. The influenza vaccine kit of claim 1, further comprising a pharmaceutically acceptable carrier selected from the group consisting of one or more of the following: solvent, emulsifier (emulsifier) ), suspending agent, decomposer, binding agent, excipient, stabilizing agent, chelating agent, diluent (diluent), glue A gelling agent, a preservative, a lubricant, a surfactant, and an adjuvant. The influenza vaccine kit according to any one of claims 1 to 8, which is prepared by the group consisting of one or more of the following dosage forms: injection type, oral type, surface administration type And inhalation type. A method of using the influenza vaccine kit according to any one of claims 1 to 8, wherein the influenza vaccine kit is applied to a body and comprises the steps of: a. administering to the individual a first vaccine composition; and b. administering a second vaccine composition to the individual within about 1 to 10 days after administration of the first vaccine composition. The method for using the influenza vaccine kit according to claim 10, further comprising pre-administering the individual to the individual before the step a One step of the tetravalent influenza vaccine. The method of using the influenza vaccine kit of claim 11, wherein the tetravalent influenza vaccine is pre-administered to the individual about at least two weeks before the step a is performed. The method for using the influenza vaccine kit according to claim 10, wherein the tetravalent influenza vaccine comprises: an influenza A virus of the H1N1 type, an influenza A virus of the H3N2 type, and a Victorian type (Victoria). Lineage type B influenza virus and a Yamagata lineage type B influenza virus. The method for using the influenza vaccine kit according to claim 13, wherein the H1N1 influenza A virus, the H3N2 influenza A virus, and the Victoria lineage are used in the tetravalent influenza vaccine. The doses of the influenza B virus and the Yamagata lineage influenza B virus are 1 to 30 μg, respectively. The method for using the influenza vaccine kit according to claim 13, wherein the H1N1 influenza A virus strain: A/California/7/2009 virus or A/California/7/2009 (H1N1) pdm09 Class of virus (A/California/7/2009 (H1N1) pdm09-like virus). The method of using the influenza vaccine kit according to claim 12, wherein the H3N2 influenza A virus A/Texas/50/2012 virus or virus antigenicity is similar to the cell proliferation prototype A/Victoria/ 361/2011 type of virus. The method of using the influenza vaccine kit according to claim 13 of the patent application, wherein the Victoria lineage type B influenza disease Virus B/Brisbane/60/2008 virus. The method of using the influenza vaccine kit according to claim 13, wherein the Yamagata lineage influenza B virus is a B/Massachusets/2/2012 virus.