HK1236817B - Rhinovaccination system of influenza vaccine - Google Patents

Description

Flu vaccine nasal delivery system

Technical Field

The present invention relates to a transnasal vaccination system for use in conjunction with a medical syringe for administering an influenza vaccine composition to the nasal mucosa.

Background Art

Influenza is an acute respiratory infection caused by the influenza virus, particularly prevalent during the winter months. Furthermore, influenza can sometimes cause widespread outbreaks and even lead to fatalities. Flu vaccination is known to be effective in preventing influenza, and it can be widely administered before the epidemic season.

The only influenza vaccine licensed in Japan consists of inactivated protein components of influenza virus antigens administered subcutaneously. Currently, split vaccines are used as seasonal influenza vaccines. While these vaccines are highly effective in preventing severe influenza, such as pneumonia, they lack sufficient protection against infection due to low antibody induction activity in the upper respiratory tract mucosa, the site of influenza virus infection. Furthermore, these injections pose a risk of side effects such as pain and inflammation caused by localized administration.

Numerous trials have been conducted to address these issues with influenza vaccination, with nasal vaccines attracting attention due to their novel delivery method. However, despite being widely used in clinical practice, split vaccines, when administered directly nasally to experimental animals and humans, fail to induce a high immune response against influenza viruses.

Against this backdrop, the world's first intranasal split influenza vaccine using Escherichia coli heat-labile enterotoxin as an adjuvant was licensed in Switzerland (manufactured by Berna Biotech (Switzerland) under the trade name Nasalflu) and launched in October 2000. However, clinical use was discontinued in February 2004 due to adjuvant toxicity. Furthermore, Patent Document 1 discloses an adjuvanted intranasal influenza vaccine, demonstrating that the use of an adjuvant can enhance immune induction. However, concerns remain regarding adjuvant toxicity in practical applications.

For nasal administration, the complex structure of the nasal cavity must also be considered, and it is expected that the influenza vaccine will diffuse widely in the nasal cavity, adhere to the nasal cavity, and remain there for a long time. For example, Patent Document 2 discloses that a base can be used for spray administration.

Pump sprayers, such as the airless sprayer used in Patent Document 2, can achieve good formulation spray applicability (spray dispersibility and formulation particle size uniformity, etc.), thereby being expected to obtain the desired sufficient drug effect. However, due to structural reasons in the spray container, it is difficult to just encapsulate a single administration dose, and thus it is difficult to make a disposable drug delivery system. Therefore, in order to use this pump sprayer for nasal administration of influenza vaccine, an excess of vaccine preparation can only be encapsulated in the spray container. After administration to one person, the remaining preparation is discarded together with the sprayer, or the remaining preparation is shared by multiple people to save costs. However, the nozzles of this airless sprayer are inserted into the nasal cavities of multiple patients or vaccinees respectively, which makes most people feel psychologically repulsive. Moreover, this use is unhygienic and has the risk of causing other infectious diseases (hospital infection).

As mentioned above, there is currently great hope for the development and implementation of a nasal influenza vaccine as a next-generation influenza vaccine, replacing traditional influenza vaccines administered subcutaneously or intramuscularly. However, practical applications present numerous challenges, such as reducing the toxicity of adjuvants used to enhance immune induction and designing drug delivery devices to enhance their effectiveness.

Prior art literature

Patent Literature

[Patent Document 1] WO 2010/114169

[Patent Document 2] WO 2007/123193

Summary of the Invention

Problems to be solved by the invention

One of the purposes of the present invention is to provide a delivery system for an influenza vaccine composition for nasal spray administration. The influenza vaccine is prepared from whole inactivated influenza virus particles as a licensed antigen and does not contain an adjuvant. Despite having a low antigen content, it exhibits high efficiency and low side effects, and is used in combination with a delivery device.

In addition, the related application of the present application has been published, and its international publication number is WO2014/103488. However, the priority date of the present application is earlier than the publication date of the related application, so the related application should not be regarded as prior art of the present application.

Means of solving the problem

The inventors of this application conducted extensive research to address the above-mentioned issues and discovered that a composition comprising (i) a colloid base for transnasal administration comprising a carboxyvinyl polymer treated with external shear force to impart spray properties and (ii) inactivated whole influenza virus particles can enhance human immunogenicity without the use of an adjuvant. Furthermore, they developed a drug delivery system by packaging this composition into a metered-dose syringe-type dispenser with a nozzle having an optimal shape and structure. This new discovery led to the completion of the present invention. The present invention can provide the following examples.

[1] A nasal influenza vaccine administration system comprising a syringe-type ejector filled with an influenza vaccine composition; the influenza vaccine composition comprises (i) inactivated whole influenza virus particles and (ii) a colloid base containing a carboxyvinyl polymer and treated with external shear force to impart spray properties, and the colloid base is characterized by being adjuvant-free.

[2] The influenza vaccine transnasal inoculation system according to [1], wherein the syringe-type ejector is a medical syringe equipped with a nasal spray nozzle and having a front end opening in fluid communication with the syringe barrel, wherein the nasal spray nozzle comprises:

a hollow nozzle body having a front end portion on which a nozzle ejection hole is formed,

a solid filler rod disposed in the nozzle body, and

A nozzle cavity is formed between the filling rod and the nozzle body to achieve fluid communication between the front end opening and the nozzle ejection hole,

The diameter of the nozzle ejection hole is in the range of 0.25 mm to 0.30 mm.

[3] The influenza vaccine transnasal vaccination system according to [1] or [2], wherein the amount of the inactivated whole influenza virus particles (i) is 1-500 μg HA/mL per influenza virus strain.

[4] The influenza vaccine transnasal inoculation system according to any one of [1] to [3], wherein the influenza vaccine composition contains 0.1 w/v% to 1.0 w/v% of a carboxyvinyl polymer.

[5] The influenza vaccine nasal inoculation system according to any one of [1] to [4], wherein the spray performance is to control (1) the particle size distribution of the composition forming the spray, (2) the uniformity of the spray density and/or (3) the spray angle.

[6] The influenza vaccine transnasal inoculation system according to any one of [1] to [3], wherein the influenza vaccine composition is prepared by: applying an external shear force to a colloidal base comprising 0.5 w/v% to 2.0 w/v% of a carboxyvinyl polymer to control (1) the particle size distribution of the composition forming the spray, (2) the uniformity of the spray density, and/or (3) the spray angle as spray properties, thereby obtaining the colloidal base; and then

The obtained colloidal base is uniformly mixed with a virus stock solution containing inactivated whole influenza virus particles in a short time under stress-free conditions.

[7] The influenza vaccine transnasal inoculation system according to any one of [1] to [6], wherein the influenza vaccine composition is prepared by using a colloidal base containing a carboxyvinyl polymer and subjected to an external shearing force treatment to impart a spraying property, wherein the spraying property is controlled as follows: (1) in terms of the particle size distribution of the composition forming the spray, the average particle size is within the range of 30 μm to 80 μm and the proportion of particles distributed within the range of 10 μm to 100 μm is 80% or more;

(2) The spray density is uniform, thus forming a homogeneous full cone, and

(3) The spray angle is adjusted within the range of 30°-70°.

[8] The influenza vaccine transnasal inoculation system according to any one of [1] to [6], wherein the influenza vaccine composition is prepared by using a colloidal base containing a carboxyvinyl polymer and subjected to an external shearing force treatment to impart a spraying property, wherein the spraying property is controlled such that (1) the average particle size of the composition forming the spray is within the range of 40 μm to 70 μm and the proportion of particles distributed between 10 μm and 100 μm is 90% or more,

(2) The spray density is uniform, thus forming a homogeneous full cone.

(3) The spray angle is adjusted within the range of 40°-60°.

[9] The influenza vaccine transnasal inoculation system according to any one of [2] to [8], wherein the nozzle ejection hole does not substantially contain a curved portion.

[10] The influenza vaccine transnasal inoculation system according to any one of [2] to [9], wherein the thickness of the front end portion where the nozzle ejection hole is formed is in the range of 0.20 mm to 0.30 mm along the ejection direction of the preparation.

[11] The influenza vaccine transnasal inoculation system according to any one of [2] to [10], wherein at least a portion of the inner wall of the nozzle body is formed into a cylindrical shape, and at least a portion of the outer wall of the filling rod is formed into a cylindrical shape having a plurality of grooves spaced apart in the circumferential direction.

the nozzle cavity being formed between the at least a portion of the inner wall of the nozzle body and the at least a portion of the outer wall of the fill rod; and

The filling rod includes a vortex forming member disposed opposite the front end portion of the nozzle body.

[12] The influenza vaccine transnasal inoculation system according to [11], wherein, in the vortex forming member, the flow direction of the preparation from the groove of the filling rod deviates from the central axis, thereby forming a vortex of the preparation.

[13] The influenza vaccine nasal inoculation system according to [11] or [12], wherein at least a portion of the inner wall of the nozzle body is formed so that a cross section within a plane substantially perpendicular to the ejection direction continuously or in stages decreases toward the ejection direction.

Effects of the Invention

The present invention provides an influenza vaccine composition comprising inactivated whole influenza virus particles as an active ingredient but without an adjuvant, which can induce a high immune response even with a small amount of antigen. Moreover, since the composition does not contain an adjuvant, side effects are minimal. Furthermore, by using a dosing system equipped with a metered-dose syringe-type dispenser having an optimally shaped nasal spray nozzle, the influenza vaccine composition can be suitably used to respond to influenza epidemics.

The influenza vaccine composition of the present invention can diffuse widely in the nasal mucosa and retain for a long time because the composition contains a colloidal base containing a carboxyvinyl polymer and is treated by applying external shear force to impart spray properties. Therefore, the influenza vaccine composition of the present invention can induce a high immune response with a small amount of antigen.

According to the preparation process of the influenza vaccine composition of the present invention, since the virus particles are processed in a short time without applying stress, the provided influenza vaccine composition well maintains the antigenicity of the inactivated whole virus particles and can induce a high immune response with few side effects.

Although the present invention does not contain an adjuvant as an immunopotentiator, it can provide immune induction equivalent to or stronger than that of a composition containing an influenza virus vaccine and an adjuvant to the upper respiratory tract mucosa and the whole body.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is a partially broken side view of the overall structure of a medical syringe including a nasal spray nozzle according to one embodiment of the present invention.

2( a ) and 2 ( b ) are partially cutaway perspective views of the overall structure of a nasal spray nozzle according to an embodiment of the present invention, respectively showing the configurations before and after the filling rod is inserted into the nozzle body.

Figure 3(a) is a vertical cross-sectional view of the nasal spray nozzle of Figure 2(b), and Figures 3(b) to 3(d) are horizontal cross-sectional views of the nasal spray nozzle when viewed along lines B-B, C-C, and D-D in Figure 3(a), respectively.

4( a ) and 4 ( b ) are enlarged cross-sectional views of the front end portion of the nozzle body, wherein the front end portion of FIG. 4( a ) is provided with a bent portion, and the front end portion of FIG. 4( b ) is not provided with a bent portion.

FIG5 shows the results of measuring the particle size distribution of the preparation of Example 4 sprayed using the syringe-type ejector of the present invention using a laser diffraction particle size analyzer.

6 shows the measurement using a high-speed microscope of the spray angle of the preparation sprayed from the front end of the syringe-type ejector of Example 4 of the present invention. The spray angle of the sprayed preparation was 52.27°.

7 shows the spray behavior of the preparation sprayed from the syringe-type ejector of the present invention in Example 4, measured using a spray-type test paper. The result is a uniform full cone.

DETAILED DESCRIPTION

The present invention provides a nasal influenza vaccine inoculation system, comprising:

A medical syringe filled with an influenza vaccine composition, the medical syringe having a front end opening in fluid communication with a syringe barrel and equipped with a nasal spray nozzle, the nasal spray nozzle comprising: a hollow nozzle body having a front end portion with a nozzle ejection hole formed thereon, a solid filling rod disposed in the nozzle body, and a nozzle cavity formed between the filling rod and the nozzle body for achieving fluid communication between the front end opening and the nozzle ejection hole, wherein the nozzle ejection hole has a diameter within a range between 0.25 mm and 0.30 mm.

The influenza vaccine composition comprises a colloid base containing a carboxyvinyl polymer and treated by applying external shear force to impart nebulization properties, and inactivated whole influenza virus particles, and is characterized by being adjuvant-free.

As used herein, the term "colloid base containing a carboxyvinyl polymer and treated with external shear force to impart sprayability" refers to the "colloid base containing a skin/mucosal adhesive," as disclosed in WO 2007/123193. This base comprises a carboxyvinyl polymer and, optionally, gellan gum, whose viscosity is adjusted by applying external shear force. This base is characterized in that its viscosity can be adjusted to various viscosities by applying external shear force, thereby enabling control of the spray diffusion angle and spray density from the spray container to meet specific objectives. Furthermore, a drug delivery system using a metered-dose syringe-type sprayer equipped with an optimally shaped nasal spray nozzle of the present invention can achieve superior formulation sprayability (spray dispersibility and formulation particle size uniformity, etc.), similar to that achieved using a pump sprayer (such as the airless sprayer disclosed in WO 2007/123193). This allows for a wider and longer-term diffusion of inactivated flu-like virus particles within the nasal mucosa, thereby enhancing the immunogenicity of the antigen.

The carboxyvinyl polymer as a raw material component of the colloid base of the present invention is a hydrophilic polymer obtained by polymerizing acrylic acid as a main component and can be selected from pharmaceutical additives generally used for preparing aqueous colloids without limitation.

The colloid base containing a carboxyvinyl polymer and treated by applying an external shear force to impart spraying properties has a carboxyvinyl polymer content of 0.1-1.0 w/v%, preferably 0.3-0.7 w/v%.

The vaccine of the present invention is characterized by containing inactivated whole influenza virus particles as antigens. As used herein, inactivated whole influenza virus particles are virions produced by purifying a virus suspension obtained by culturing influenza virus under conditions that maintain the virus's morphology. Therefore, the influenza vaccine of the present invention excludes split vaccines containing subvirions and subunit vaccines containing purified HA or NA, and is also referred to as a whole virus vaccine.

The inactivated whole influenza virus particles are preferably obtained by purifying a virus suspension that does not contain surfactants and ethers. The virus stock solution used herein refers to a virus solution containing inactivated whole influenza virus particles, which is purified or concentrated and then mixed with the colloidal base of the present invention. In the vaccine of the present invention, the concentration of inactivated whole influenza virus particles is preferably 1-500 μg HA/mL (converted as HA) per vaccine virus strain, and more preferably 20-250 μg HA/mL (converted as HA). This concentration can be obtained by measuring the concentration of HA protein.

Influenza virus used here comprises all virus types and subtypes thereof of currently known influenza virus and all virus types and subtypes thereof separated and identified in the future.In addition, although large-scale infection in the human race has not been observed so far, from the viewpoint of the necessity that may be prevalent in the human race in the future, it is preferably selected from the influenza A virus subtype of the group consisting of the H1-H16 subtype that does not comprise H1 and H3 subtypes (that is, H2 and H4-H16) and the combination of the influenza A virus subtype of the group consisting of the N1-N9 subtype. These subtypes are also referred to as novel influenza virus. In the aforementioned subtype, it is preferably: the combination of the subtype of the group consisting of the H5, H7 and H9 subtypes and the subtype of the group consisting of the N1-N9 subtype. Influenza virus can be from a kind of virus strain, or from two or more virus strains belonging to the same subtype, or from two or more virus strains belonging to different subtypes.

Influenza virus used herein includes virus strain that separates from infected animals or people and recombinant virus that is constructed in cultured cells by gene method.For the culture method of influenza virus, virus can be inoculated in the allantoic cavity of chicken egg and cultured, or cultured cells are cultured after infection.

Adjuvants are a general term for substances that have the ability to regulate immune responses (such as enhancement and inhibition), and are used as immunopotentiators added to vaccines to increase the immunogenicity of antigens. A variety of adjuvants have been studied so far. The use of adjuvants improves the immune effect of vaccines, but has the disadvantage of producing side effects such as inflammation. Of course, there are several adjuvants available as alternatives for nasal vaccines, but since there is no adjuvant with broad safety, no nasal vaccine containing an adjuvant has yet to be approved.

The inventors have discovered that by using a colloidal base with the aforementioned excellent spray properties (such as high adhesion to the nasal mucosa) in the aforementioned whole-virus vaccine, a highly effective vaccine with fewer side effects can be prepared with a smaller amount of antigen without the use of an adjuvant. In addition, the inventors have also discovered that with the help of a device that can even spray a high-viscosity colloidal base, the influenza vaccine composition can be sprayed onto the nasal mucosa, and the average particle size of the resulting spray-forming composition is within a suitable range of 30 μm-80 μm (preferably within the range of 40 μm-70 μm), and the particle size distribution between 10 μm and 100 μm is more than 80% (preferably more than 90%). The spray angle from the device is set in the range of 30°-70° (preferably in the range of 40°-60°), so that the composition can be administered to the desired location in the nasal cavity and the spray density is uniform, thereby forming a homogeneous full cone. The inventors have also discovered a process and method for using the composition to prevent influenza. Based on the above new findings, the present invention has been completed.

The vaccine of the present invention, in addition to inactivated whole influenza virus particles and a colloidal base, may also contain a pharmaceutically acceptable carrier. The carrier used herein can be any carrier commonly used in the preparation of vaccines and intranasal formulations, including, for example, saline, buffered saline, glucose, water, glycerol, isotonic aqueous buffer solutions, and combinations thereof. Furthermore, the vaccine of the present invention may optionally contain a preservative (e.g., sodium thiomersalate), an isotonicity agent, a pH adjuster, a surfactant, and an inactivating agent (e.g., formalin).

The vaccine of the present invention is intended for spray administration into the nasal cavity.

The vaccine of the present invention can prevent influenza or alleviate influenza symptoms.

Regarding the method of administering the vaccine, the spray may be delivered to a single nostril or both nostrils using the optimized nasal spray nozzle of the present invention which can be used as a disposable device.

The dosage of the vaccine should be determined based on the patient's age, sex, weight or other factors. In practice, the vaccine is usually administered at 1 μg-150 μg HA per vaccine virus strain, calculated as antigen. Preferably, 5 μg-50 μg HA is administered per vaccine virus strain.

With reference to the accompanying drawings, an embodiment of a nasal spray nozzle for use in a metered-dose syringe-type dispenser including a nasal spray nozzle according to the present invention will be described below. In the following description, directional terms (such as front, back, near, and far) are generally used for ease of understanding and are not intended to limit the present invention. Furthermore, identical components are designated by the same reference numerals throughout the drawings.

[Medical syringe]

FIG1 is a partially cutaway side view of a medical syringe 1 including a nasal spray nozzle 10 according to one embodiment of the present invention. As shown in FIG1 , the medical syringe 1 generally comprises a syringe body 4 made of synthetic resin or glass and having a syringe barrel 3 capable of storing a pharmaceutical preparation, and a plunger rod 5 inserted into the syringe barrel 3 of the syringe body 4. The medical syringe 1 also comprises a piston 7 having a fixing member 5a disposed at the distal end of the plunger rod 5 and sliding within the syringe barrel 3 to pump the preparation in the syringe barrel 3 out of the distal front opening of the syringe body 4; a grip 8 disposed around the proximal end of the syringe body 4; and a plunger end member 9 for transmitting force applied by an operator (e.g., a physician) to the plunger rod 5. The medical syringe 1 may be similar to the metered-dose syringe-type dispenser disclosed in WO 2013/145789.

It should be noted that the nasal spray nozzle 10 of the present invention is applicable to any type of medical syringe 1 that pumps a formulation from a syringe barrel 3 by pushing the plunger rod 5 (and piston 7). Therefore, the present invention is not limited to medical syringes of known construction. Therefore, this disclosure omits a detailed structural description of the medical syringe (nasal metered dose syringe-type ejector) 1 and instead provides a more detailed description of the structure and function of the nasal spray nozzle 10 used therein. It should also be noted that the disclosure of WO2013/145789 is incorporated herein by reference.

[Nasal spray nozzle]

As shown in Figure 1, the medical syringe 1 also includes a nasal spray nozzle 10 positioned opposite the front opening 6 of the syringe body 4, and a protective cap 50 for protecting the sterilized front end 22 of the nasal spray nozzle 10 from contaminants and mechanical impact. Figures 2(a) and 2(b) are partially cutaway perspective views illustrating the general structure of the nasal spray nozzle 10 according to one embodiment of the present invention. As shown, the nasal spray nozzle 10 generally includes a hollow nozzle body 20 having a front end 22 formed with a nozzle ejection hole 21, and a solid filling rod (filling rod) 30 disposed within the nozzle body 20. Figures 2(a) and 2(b) respectively illustrate the nozzle body 20 before and after the filling rod 30 is installed or inserted into the nozzle body 20. The front end 22 of the nozzle body 20 is circular and has the nozzle ejection hole 21 at its center.

Figure 3(a) is a vertical cross-sectional view of the nasal spray nozzle 10 shown in Figure 2(b). Figures 3(b), 3(c), and 3(d) are horizontal cross-sectional views of the nasal spray nozzle 10, taken along lines B-B, C-C, and D-D, respectively, in Figure 3(a). The hollow nozzle body 20 defines a substantially cylindrical interior space 24. As shown in Figures 3(c) and 3(d), this interior space 24 includes a nozzle narrow portion 25 located adjacent to the nozzle ejection orifice 21 of the hollow nozzle body 20, a nozzle wide portion 26 positioned opposite the front end opening 6 of the syringe body 4, and a nozzle shoulder 27 formed by continuously or stepwise decreasing diameter from the nozzle wide portion 26 toward the nozzle narrow portion 25.

On the other hand, the outer wall 33 of the solid filling rod 30 inserted into the nozzle body 20 is constructed to substantially complement the inner wall 23 (internal space 24) of the nozzle body 20. As shown in Figures 2(a), 3(c), and 3(d), the rod thin section 35 and the rod thick section 36 include a rod shoulder 37, the diameter of which decreases continuously or in stages from the rod thick section 36 toward the rod thin section 35.

Preferably, as shown in FIG3( a ), the inner wall 23 of the nozzle body 20 is provided with a protrusion 23 a, and the outer wall 33 of the filling rod 30 is provided with a recess 33 a for accommodating the protrusion 23 a. When the filling rod 30 is fully inserted into the interior space 24 of the nozzle body 20, the protrusion 23 a fits tightly into the recess 33 a, thereby ensuring the connection between the filling rod 30 and the nozzle body 20.

As shown in Figures 2(a)-2(b) and 3(a)-3(d), the filling rod 30 includes a plurality of grooves 38 and 39 spaced circumferentially on the thin rod portion 35 and the thick rod portion 36. Furthermore, the filling rod 30 is inserted into the nozzle body 20, thereby forming a gap 40 between the nozzle shoulder 27 and the rod shoulder 37 (Figure 3(a)). Therefore, the nasal spray nozzle 10, as shown in the assembled Figure 2(b), has a nozzle cavity 42 defined by the grooves 38 and 39 and the gap 40, establishing fluid communication. Specifically, the preparation 2 dispensed from the front opening 6 of the syringe body 4 passes through the nozzle cavity 42 and reaches the front end 22 of the nasal spray nozzle 10.

Furthermore, as shown in FIG3(b), the filling rod 30 includes a vortex-forming member 44 positioned opposite the front end 22 of the nasal spray nozzle 10. This vortex-forming member 44 is configured to create a vortex in the preparation 2 flowing from the grooves 38 of the rod's thin-diameter portion 35 before being ejected from the nozzle orifice 21 of the nozzle body 20. In particular, the end of the rod's thin-diameter portion 35, which constitutes the vortex-forming member 44, extends in a direction offset from the vertical center axis of the nozzle orifice 21. This vortex-forming member 44 increases the spray angle of the preparation 2 before it is ejected from the nozzle orifice 21, thereby ensuring a more uniform spray of the preparation 2.

As shown in Figures 3(c)-3(d), the grooves 38 of the thin stem portion 35 are preferably designed to be fewer in number than the grooves 39 of the thick stem portion 36, thereby increasing the pressure of the preparation 2 within the vortex-forming member 44 before spraying from the nozzle outlet 21. Furthermore, because the diameters of the thick stem portion 36 and the thin stem portion 35 decrease continuously or in stages from the thick stem portion 36 toward the thin stem portion 35, the nasal spray nozzle 10 can be easily inserted deep into the patient's nasal cavity, allowing the preparation to be sprayed to the patient's inferior turbinate and even deeper. Therefore, the diameter of the thin stem portion 35 is preferably sufficiently smaller than the patient's nostrils to avoid any fear.

Examples

According to the following method, a colloidal base and three virus stock solutions were prepared, and the two were mixed as follows to prepare the influenza vaccine composition shown in the Examples. Each viscosity was measured at 20°C using an E-type viscometer.

Preparation of colloid base

Colloidal base example 1

<Preparation of Virus Stock Solution Containing Inactivated Whole Influenza Virus Particles>

Virus stock solution example 1

Virus stock solution example 2

Virus stock solution example 3

<Mixture of colloid base and virus stock solution>

Each of the above-mentioned colloidal base example (1) and viral stock solution examples (1)-(3) was stirred and mixed in a ratio of 1:1 to prepare homogeneous influenza vaccine composition examples 1, 2 and 3, respectively. The composition of the composition of each example and the physical properties/spray performance exhibited when sprayed using a spray device or a syringe-type ejector are shown below. The stirring and mixing process can be completed gently in a short time and does not exert stress on the inactivated whole virus antigen. In the resulting influenza vaccine composition, the amount of each component and the spray performance exhibited by spraying the composition using an appropriate device are shown below.

Example 1

Example 2

Example 3

Example 4

Alternatively, the inactivated whole antigen used in the above examples was used to prepare influenza vaccine compositions without a colloid base according to the composition in the following table, as Comparative Examples 1-4.

Comparative Example 1

Comparative Example 2

Comparative Example 3

Comparative Example 4

Tests to assess immune response (1)

The influenza vaccine compositions prepared in Example 1 and Comparative Example 1 were administered via nasal spray using appropriate disposable devices to two groups of four adult volunteers, each receiving 0.25 mL of HA per nostril (45 μg HA for both nostrils combined). The inoculations were administered twice, three weeks apart.

Then, blood and nasal wash fluid were collected continuously to determine and analyze the neutralizing antibody titers of the vaccine virus strains. The results of Example 1 are shown in Table 1, and the results of Comparative Example 1 are shown in Table 2.

Table 1

Table 2

Comparisons of the vaccine of Example 1 (viral stock solution + colloid base) with the vaccine of Comparative Example 1 (a composition containing inactivated split influenza virus antigens but no colloid base) showed no increase in neutralizing antibody titers in the serum of three-quarters of the subjects vaccinated with the Comparative Example 1 vaccine, while four-quarters of the subjects vaccinated with the Example 1 vaccine showed a significant increase in neutralizing antibody titers in the serum. Neutralizing antibody titers in the nasal wash fluid of all subjects vaccinated with both the Example 1 and Comparative Example 1 vaccines increased, but the increase was greater with the Example 1 vaccine.

Tests to assess immune response (2)

The influenza vaccine compositions prepared in Example 2 and Comparative Example 2 were administered by nasal spray inoculation using an appropriate disposable device. The Example 2 vaccine group consisted of 25 adult volunteers, and the Comparative Example 2 vaccine group consisted of 24 adult volunteers. The single nostril inoculation volume was 0.25 mL (45 μg HA in both nostrils combined), and the inoculations were administered twice, three weeks apart, and then again approximately six months later, for a total of three doses.

Three weeks after the third vaccination, blood and nasal wash fluid were collected to measure and analyze the neutralizing antibody titers against the vaccine virus strain. The results are shown in Table 3.

Table 3

The vaccine of Example 2 (virus stock solution + colloid base) was compared with the vaccine of Comparative Example 2 (containing only virus stock solution). The results showed that the immune reactivity of the vaccine of Example 2 containing the colloid base was greatly improved compared with the vaccine of Comparative Example 2.

It is previously known that humans who have never been exposed to influenza virus antigens (e.g., infants/children) have a low immune response. It is conceivable that the immune response of these susceptible populations to influenza vaccines can be evaluated by assessing the immune response of healthy adults vaccinated with highly pathogenic avian influenza virus (H5N1 strain), since most healthy adults have never been exposed to this avian influenza virus (i.e., are in a naive state).

As shown in the above results, it was found that even for susceptible people, high levels of neutralizing antibody titers in serum and nasal washes could be induced by nasal inoculation of the vaccine of Example 2 (virus stock solution + colloid base) three times.

Immune response analysis test (3)

The influenza vaccine compositions prepared in Example 3, Comparative Example 3, and Comparative Example 4 were administered nasally using a syringe-type dispenser. The Example 3 vaccine group consisted of 47 adult volunteers, and the Comparative Example 3 vaccine group also consisted of 47 adult volunteers. Each nostril was inoculated with 0.25 mL (15 μg HA/strain/0.5 mL for both nostrils), with a total of two inoculations three weeks apart. Additionally, a test group consisting of 38 adult volunteers received a single subcutaneous inoculation of 0.5 mL (15 μg HA/strain/0.5 mL) of the influenza vaccine composition prepared in Comparative Example 4 (the current vaccine).

Three weeks after the last vaccination (the second and first doses, respectively), blood and nasal washes were collected to measure and analyze neutralizing antibody titers against the vaccine strains. The results for the different influenza vaccine types are shown in Tables 4 and 5, respectively.

Table 4

Table 5

Comparison of the results of nasal administration of the vaccine of Example 3 (viral stock solution + colloid base), nasal administration of the vaccine of Comparative Example 3 (containing only viral stock solution), and subcutaneous administration of the vaccine of Comparative Example 4 (currently used subcutaneous vaccine) showed that nasal administration of the colloid-based vaccine of Example 3 significantly increased the immune reactivity of the vaccine compared to nasal administration of the vaccine of Comparative Example 3. Furthermore, analysis of nasal wash fluids showed that the nasal administration of the vaccine of Example 3 induced neutralizing antibodies in the nasal mucosa, while the subcutaneous administration of the vaccine of Comparative Example 4 (currently used vaccine) did not show such an induction effect.

In summary, a nasal influenza vaccine nasal injection system was prepared by filling a medical syringe with the nasal influenza vaccine formulation using the colloid base treated with external shear force in Example 4, wherein the medical syringe has a front opening in fluid communication with the syringe barrel, and is equipped with a nasal spray nozzle including a hollow nozzle body (having a front end portion with a nozzle ejection hole formed thereon), a solid filling rod within the nozzle body, and a nozzle cavity formed between the filling rod and the nozzle body to achieve fluid communication between the front opening and the nozzle ejection hole, wherein the nozzle ejection hole has a diameter within a range between 0.25 mm and 0.30 mm;

The spray performance of the system is that it can be controlled so that: (1) the particle size distribution of the composition forming the spray, that is, the average particle size is in the range of 30μm-80μm (e.g., 59.6μm), and the distribution ratio of particles within 10μm-100μm is greater than 80% (e.g., 85.6%); (2) the spray density is uniform to form a homogeneous full cone; and (3) the spray angle is adjusted within the range of 30°-70° (e.g., 52.27°).

Reference numerals:

1: Medical syringe, 2: Pharmaceutical preparation, 3: Syringe barrel, 4: Syringe body, 5: Piston rod, 5a: Fixing part, 6: Opening, 7: Piston, 8: Hand grip, 9: Piston end member, 10: Nasal spray nozzle, 20: Nozzle body, 21: Nozzle ejection hole, 22: Front end, 23: Inner wall, 23a: Protrusion, 24: Internal space, 25: Nozzle thin diameter part, 26: Nozzle thick diameter part, 27: Nozzle shoulder, 30: Filling rod, 33: Outer wall, 33a: Recess, 35: Rod thin diameter part, 36: Rod thick diameter part, 37: Rod shoulder, 38, 39: Groove, 40: Gap, 42: Nozzle cavity, 44: Vortex forming member, 46: Bend, 50: Protective cover.

Claims (13)

1. A nasal influenza vaccine administration system comprising a syringe-type sprayer filled with an influenza vaccine composition; said influenza vaccine composition comprising (i) inactivated whole influenza virus particles and (ii) a colloidal base containing a carboxyl vinyl polymer and subjected to external shear force to impart spray properties, and said colloidal base being characterized as being adjuvant-free. 2. The nasal inoculation system for influenza vaccines according to claim 1, wherein the syringe-type sprayer is a medical syringe equipped with a nasal spray nozzle and having a front opening in fluid communication with the syringe barrel, the nasal spray nozzle comprising: A hollow nozzle body having a front end portion on which a nozzle ejection orifice is formed. A solid filling rod disposed in the nozzle body, and A nozzle cavity is formed between the filling rod and the nozzle body to achieve fluid communication between the front opening and the nozzle discharge orifice. The nozzle orifice diameter ranges from 0.25mm to 0.30mm. 3. The nasal inoculation system for influenza vaccines according to claim 1 or 2, wherein the amount of the inactivated whole influenza virus particles (i) is 1-500 μg HA/mL per influenza virus strain. 4. The nasal inoculation system for influenza vaccine according to claim 1 or 2, wherein the influenza vaccine composition contains 0.1 w/v% to 1.0 w/v% of a carboxyvinyl polymer. 5. The nasal inoculation system for influenza vaccine according to claim 1 or 2, wherein the spray performance is characterized by controlling (1) the particle size distribution of the composition forming the spray, (2) the uniformity of the spray density and/or (3) the spray angle. 6. The nasal inoculation system for influenza vaccines according to claim 1 or 2, wherein the preparation of the influenza vaccine composition is achieved by: applying an external shear force to a colloidal base comprising 0.5 w/v%-2.0 w/v% of a carboxyvinyl polymer to control (1) the particle size distribution of the composition forming the spray, (2) the uniformity of the spray density, and/or (3) the spray angle as spray properties, thereby obtaining the colloidal base; then The obtained colloidal base was uniformly mixed with a viral stock solution containing inactivated whole influenza virus particles in a short time under stress-free conditions. 7. The nasal inoculation system for influenza vaccine according to claim 1 or 2, wherein the influenza vaccine composition is prepared by using a colloidal base agent containing a carboxyl vinyl polymer and subjected to external shear force to impart spray properties, wherein the spray properties are controlled such that (1) in terms of the particle size distribution of the composition forming the spray, the proportion of average particle size in the range of 30 μm to 80 μm and particle size distribution in the range of 10 μm to 100 μm is 80% or more. (2) The spray density is uniform, thus forming a homogeneous full cone shape, and (3) The spray angle is adjusted within the range of 30°-70°. 8. The nasal inoculation system for influenza vaccine according to claim 1 or 2, wherein the influenza vaccine composition is prepared by using a colloidal base agent containing a carboxyl vinyl polymer and subjected to external shear force to impart spray properties, wherein the spray properties are controlled such that (1) in terms of the particle size distribution of the composition forming the spray, the proportion of average particle size in the range of 40 μm-70 μm and particle size distribution in the range of 10 μm-100 μm is 90% or more. (2) The spray density is uniform, thus forming a homogeneous full cone shape, and (3) The spray angle is adjusted to the range of 40°-60°. 9. The nasal inoculation system for influenza vaccine according to claim 2, wherein the nozzle orifice substantially does not contain a bend. 10. The nasal inoculation system for influenza vaccine according to claim 2, wherein the thickness of the front end having the nozzle orifice along the spray direction of the preparation is in the range of 0.20 mm to 0.30 mm. 11. The nasal inoculation system for influenza vaccine according to claim 2, wherein at least a portion of the inner wall of the nozzle body is formed in a cylindrical shape, and at least a portion of the outer wall of the filling rod is formed in a cylindrical shape having a plurality of grooves spaced apart in the circumferential direction. The nozzle cavity is formed between at least a portion of the inner wall of the nozzle body and at least a portion of the outer wall of the filling rod; and The filling rod includes a vortex forming portion disposed opposite to the front end of the nozzle body. 12. The nasal inoculation system for influenza vaccine according to claim 11, wherein, in the vortex forming section, the flow direction of the formulation from the groove of the filling rod deviates from the central axis, thereby forming a vortex of the formulation. 13. The nasal inoculation system for influenza vaccines according to claim 11 or 12, wherein at least a portion of the inner wall of the nozzle body is formed such that the cross-section in a plane substantially perpendicular to the ejection direction continuously or progressively decreases toward the ejection direction.