ORAL VIRAL VACCINE CARRIER

In the case of intestinal bacteria in oral virus vaccine for preventing virus infection, target field to virus vaccine has reached over long for the current must pass through the substrate. Unlike field, pH 1 to 3 represents a unit has a low pH, low pH conditions maintaining stability of virus vaccine needs to be disclosed. However, recorded and which virus vaccine is unstable in low pH so that the door number, has a small effect efficient vaccines of the virus does not have door number flow tides.

Thus, the present invention victims of the oral virus vaccine delivery door number point if the result of the study a kind, pH sensitive polymer cationic polymer and low pH under conditions oral virus vaccine particles capable of delivering to the present invention stably vaccines without damaging to confirm the arrears of work.

Thus, the present invention refers to cationic polymer layer and pH sensitive polymer layer injected into virus vaccine particles including oral virus vaccine polymer such as a number under public affairs substrate. According to one example of the present invention to also 1 shown to a manufacturing method and organization of the type oral virus vaccine delivery. In the present invention, pH sensitive polymer layer sequence in cationic polymer layer can be formed on oral virus vaccine particles.

In the present invention, the "formed on", meaning that only those in direct contact with the components formed in a layered manner and not to, the other components is further formed between between components comprising the meanings. E.g., "in a transfer liquid" as non-, number 1 component formed on the surface of the components as well as the meanings in direct contact with number 2, number 3 between the electrodes and said number 1 number 2 component component component includes the meanings.

According to one example of the present invention, oral virus vaccine particles; cationic polymer layer formed on said virus vaccine particles; and said pH sensitive polymer layer formed on cationic polymer layer including oral virus vaccine polymer such as a number under public affairs substrate.

Other according to one example of the present invention, oral virus vaccine particles; said virus vaccine particles formed on pH sensitive polymer layer; and said pH sensitive polymer layer formed on a polymer such as a cationic polymer layer including oral virus vaccine number under public affairs substrate.

In one embodiment, oral virus vaccine mean particle size of the particles, the number is a but not limited to, 0. 1 to implementation being 10um.

In the present invention, oral virus vaccine particles intestinal virus vaccines against only the number without available. For example, according to one example of the present invention oral virus vaccine be a porcine epidemic diarrhea virus particles.

Oral porcine epidemic diarrhea virus vaccine is oral vaccines for the prevention and treatment of porcine epidemic diarrhea virus only the number without available. E.g., oral porcine epidemic diarrhea virus vaccine is stipendiary cross pharmaceuticals can be purchased from. PH range of pH 7 to 8 oral administration of commercially available porcine epidemic diarrhea virus vaccine is crucial stability at ambient temperatures and, viral or pH 11 or more pH 5 hereinafter is to rapidly killing, destruction compound number exists in the flow tides. The, field to efficiently because difficult. However, said oral porcine epidemic diarrhea virus vaccine particles cationic polymer further comprises a carbide layer and polymer layer pH, low pH or pH 1 to pH 5 hereinafter 3 viral particles even in the base plate have been killed or destroyed the pin is (in the embodiment for reference).

Oral porcine epidemic diarrhea virus vaccines exhibit weak negative charge, said virus vaccine particles cationic polymer can be as an antireflective. By forming cationic polymer layer, even low pH can be contribute to maintaining the stability of virus vaccine. In addition, a serine proteinase present in cationic polymer layer field, e.g. trypsin, thus the virus vaccines such as decomposed by efficiently field can be transferred.

In one embodiment, the cationic polymer layer the number one but not limited to, poly lysine, poly piii and a selected from the group consisting at least one cationic polymer consisting of arginine may be disclosed.

In one embodiment, the cationic polymer represented by formula 1 can be represented.

[Formula 1]

K (poly - M)

In formula said, M is lysine, histidine or arginine and, k is 2 to 50 by a goniophotometer.

In one embodiment, cationic polymer layer has a thickness 1um to 1 nm, 5 nm to 500 nm, 10 nm to 300 nm, 10 nm to 200 nm, 10 nm to 100 nm, 10 nm to 90 nm, 10 nm to 70 nm, 10 nm to 50 nm, 10 nm to 30 nm, 10 nm to 20 nm or implementation being. Said range, in the range of pH 1 to pH 3 virus vaccine particles and effectively maintained stability, without immune action desired to go to delivery can be effectively virus vaccine particles. In addition, pH-sensitive polymer layer on when cationic polymer, cationic polymer layer has a thickness of said range a good-quality, pH sensitive polymer layer formed well.

In the present invention, pH of pH 1 to pH sensitive polymer layer 3 is maintained in a low frequency range structure, pH range of pH 7 to 9 ionization with each other. For example, according to pH sensitive polymer of the present invention in one example 4 pKa value. 66 and, pKb value 7. 0 are disclosed. Said pKa value lower than the pKa maintained into a hydrogen bonding polymer layer in the structure is stable in a low frequency range, said pKb higher pKa value generating anions are ionized polymer in a low frequency range and, as a result achieved on the decomposition of the polymer induced electrostatic repelling, virus vaccine to the piston to be coated. The, by pH sensitive polymer layer, a low-range of pH for inducing cancer cell death and/or destruction in virus vaccine particles simultaneously coating stability, pH conditions can be intestinal of emitting virus vaccine.

In one embodiment, the pH sensitive polymer layer characterized ionized pH 7 to 9.

In one embodiment, the pH-sensitive polymer layer the number one but not limited to, acrylate polymer can be block.

Acrylate polymer, the number is one but not limited to, poly (- co - methacrylic acid-ethyl acrylate), poly (methacrylic acid - co - methyl methacrylate) and poly (methyl methacrylate - co - methyl acrylate - co - methacrylic acid) can be one or more selected from the group consisting of. L 30 of commercially available acrylate polymer include EUDRAGIT®[...] provided 55, L 100 - 55, L 100, L12, 5, S 100, S 12, 5, FS 30 D can be using. Said acrylate polymer has a weight average molar mass (weight average molar mass) to 400000 g/mol is 100000, 100000 to 350000 g/mol, implementation being 330000 g/mol to 120000.

In one embodiment, pH-sensitive polymer layer has a thickness 1um to 1 nm, 5 nm to 500 nm, 10 nm to 300 nm, 10 nm to 200 nm, 10 nm to 100 nm, 10 nm to 90 nm, 10 nm to 70 nm, 10 nm to 50 nm, 10 nm to 30 nm, 10 nm to 20 nm or implementation being. Said range, in the range of pH 1 to pH 3 virus vaccine particles effectively maintained stability and, in the range of pH 5 to pH 9 can be effectively released particles virus vaccine. In addition, pH-sensitive polymer layer has a thickness also said when, for immune action desired to go without effectively virus vaccine particles can be delivering. In addition, these effects and pH sensitive polymer layer has a thickness of said range when the same time as a cationic polymer can be photoresist.

The present invention refers to in addition, oral virus vaccine particles and cationic polymer by mixing cationic polymer layer on oral virus vaccine particles; and said cationic polymer layer pH sensitive polymer by mixing said oral virus vaccine particles and cationic polymer to form a pH sensitive polymer layer on a manufacturing method including oral virus vaccine delivery of number under public affairs substrate.

As discussed above, pH sensitive polymer layer laminated order number is not one cationic polymer layer. Thus, the present invention refers to in addition, oral virus vaccine particles and pH sensitive polymer by mixing pH sensitive polymer layer on oral virus vaccine particles; and said pH sensitive polymer layer is formed by mixing said pH sensitive polymer layer on oral virus vaccine particles and cationic polymer cationic polymer layer manufacturing method including forming a number of oral virus vaccine delivery under public affairs substrate.

In the present invention, oral virus vaccine particles intestinal virus vaccines against only the number without available. For example, according to one example of the present invention oral virus vaccine be a porcine epidemic diarrhea virus particles. Efined porcine epidemic diarrhea virus for the discussed above.

In one embodiment, oral virus vaccine mean particle size of the particles, the number is a but not limited to, 0. 1 to implementation being 10um.

In one embodiment, pH-sensitive polymer layer formation pH 7 hereinafter, or pH 5 to pH of 6 can be carried out. PH of pH 7 to 9 of the present invention pH sensitive polymer arranged for ion, virus vaccine particles without compromising stability, pH sensitive polymer layer can be formed smoothly in the range pH, pH range of pH conditions can be said polymer layer. In addition, as well as pH polymer layer formation, cationic polymer layer formation and oral virus vaccine delivery process done in all said pH range number tank may be filled.

In one embodiment, the pH-sensitive polymer layer the number one but not limited to, acrylate polymer can be block.

Acrylate polymer, the number is one but not limited to, poly (- co - methacrylic acid-ethyl acrylate), poly (methacrylic acid - co - methyl methacrylate) and poly (methyl methacrylate - co - methyl acrylate - co - methacrylic acid) can be one or more selected from the group consisting of. L 30 of commercially available acrylate polymer include EUDRAGIT®[...] provided 55, L 100 - 55, L 100, L12, 5, S 100, S 12, 5, FS 30 D can be using. Said acrylate polymer has a weight average molar mass (weight average molar mass) to 400000 g/mol is 100000, 100000 to 350000 g/mol, implementation being 330000 g/mol to 120000.

In one embodiment, pH sensitive polymer 100 parts by weight of contrast oral virus vaccine particles 0. 5 to 20 parts by weight, 1 to 15 parts by weight, or 5 to 10 parts by weight can be mixed. Said parts by weight range, pH sensitive polymer layer of suitable thickness can be.

In one embodiment, the number one is supported by but not limited to cationic polymer, poly lysine, arginine can be one or more selected from the group consisting poly piii and poly.

In one embodiment, the cationic polymer represented by formula 1 can be represented.

[Formula 1]

K (poly - M)

In formula said, M is lysine, histidine or arginine and, k is 2 to 50 by a goniophotometer.

In one embodiment, 100 parts by weight of a cationic polymer contrast oral virus vaccine particles 0. 5 to 20 parts by weight, 1 to 15 parts by weight, or 5 to 10 parts by weight can be mixed. Said parts by weight range, cationic polymer layer of suitable thickness can be.

Hereinafter, the present invention through a in the embodiment detailed as follows. In the embodiment of the present invention is to limited to the present invention is exemplified ephemeral range are not correct.

[In the embodiment]

[Comparison example1] oral Virus vaccine delivery number of bath: virus vaccine particles -Cationic Polymer layer

30 mg 1 ml DMEM (Dulbecco Modified Eagle Medium) was reconstituted with a lysine group composed as a cationic polymer. A 600 rpm which mixed solution 1 ml stirred aqueous solution (pH 5 to 6) oral virus vaccine (PEDV; porcine epidemic diarrhea virus, be stipendiary cross pharmaceuticals) was added to 5 ml leak in small amounts. 1 hours then stirred oral virus vaccines to 600 rpm coated polymer such as a cationic polymer-configurated.

[Comparison example2] oral Virus vaccine delivery number of bath: virus vaccine particles pH sensitive polymer layer

PH sensitive polymer (Eudragit S100) 20 mg 1 ml ethanol, consisting of DMEM 1 ml every (cermets solvent) was reconstituted with quiet. A mixed solution stirred aqueous solution 5 ml 50 ul 600 rpm which oral virus vaccine (PEDV, be stipendiary cross pharmaceuticals) to leak in a small amount addition of him. 1 time to 600 rpm and then stirred ethanol solvent pH sensitive polymer coated polymer such as a stand-alone number oral virus vaccines-configurated.

[Number attainments1] oral Virus vaccine delivery number of bath: virus vaccine particles -Cationic Polymer layer - pH sensitive polymer layer

PH sensitive polymer (Eudragi S100) 20 mg 1 ml ethanol, consisting of DMEM 1 ml every (cermets solvent) was reconstituted with quiet. A comparison example 1 for the synthesis solution 5 ml to 600 rpm mixed solution 50 ul premature delivery of small amounts of a drip was added. 1 600 rpm then stirred ethanol solvent to a stand-alone time and number - - pH sensitive polymer layer oral virus vaccine particles cationic polymer layer number his oral virus vaccine polymer such as a high pressure liquid coolant. The present invention according to the number of oral virus vaccine delivery tank 1 also a predetermined level.

[Number attainments2] oral Virus vaccine delivery number of bath: virus vaccine particles - pH sensitive polymer layer -Cationic Polymer layer

30 mg 1 ml DMEM (Dulbecco Modified Eagle Medium) was reconstituted with a lysine group composed as a cationic polymer. To a stirred solution 5 ml 1 ml mixed solution delivery of comparison example 2 600 rpm which leak in a small amount addition of him. 1 - pH sensitive polymer layer - 600 rpm then stirred time oral virus vaccine particles to cationic polymer layer number his oral virus vaccine polymer such as a high pressure liquid coolant.

[Experiment example 1] stability check of oral virus vaccine delivery

The present invention according to oral virus vaccine delivery of pH according to change behavior in order to identify, using stability has been confirmed that the porcine epidemic diarrhea virus full wave rapid kit (PEDV Rapid kit).

Coating layer in pH 7 to 8 of oral PEDV (porcine epidemic diarrhea virus; porcine epidemic diarrhea virus) vaccine, vaccines and pH sensitive polymer coating layer in pH 5 to 6 of oral PEDV (comparison example 2) using only coated PEDV vaccine delivery body comparing the group, attainments number 1 and number 2 (first cationic polymer layer if the vaccine delivery) attainments (pH sensitive polymer layer if the first vaccine delivery) to a experiment group, using rapid kit, regulated and 2 of total 3 of experiment confirmed that the stability of qualifications with respect to the oral virus vaccine delivery.

Experiment conditions change in pH 1 to pH 2 to pH according to behavior to identify thereon to intestinal environment are integrated by again lowers the pH 7. 6 to 8. 5 then rapid kit based on the using stability has been confirmed.

Stability check prior to, changes in pH using pH indicator lead thiophenol therefrom. Each group of polymer such as a phenol red is contained in the DMEM pH 1 (in pH on) processing and lowering pH 8 (quantitate a pH) after response to the height, color change therefrom. As a result, as shown in fig. 2, phenol red (pH 6. 8 hereinafter yellow, pH 6. 8 to 8 in red (purple)) combined with the color change of a carrier environment altered pH conditions above and quantitate a group were not inserted.

Experiments have shown to result Rapid kit also 3. The positive control using the ray appearing in Figure 3 Control rapid kit to natural when actually substrate. PEDV TEST line is whether the presence of the line, ray appearing PEDV when identifying the presence of can.

In Figure 3, 1 once coating layer in pure oral PEDV vaccine (pH 8. 77), 2 once coating layer in oral PEDV vaccine (pH 5. 8), at a pH of 1 to 2 times 2 (low ->high) is reduced after the results of the response height 8 exhibits.

3 of the first cationic polymer poly lysine (pLys) PEDV vaccines 2 1 for a difference when coated, 3 3 1 for a pH sensitive polymer layer 2 for a difference when the difference further coating, at least two 2 3 3 (low ->high) once again based on the resulting pH 1 is divided into two the difference exhibits pH 8.

4 2 1 for a pH sensitive polymer coated PEDV vaccines where difference when, at least two 2 4 1 4 of the first cationic polymer poly lysine (pLys) difference difference when further coating, pH 1 2 4 4 (low ->high) once the difference based on the resulting pH 8 is reduced once the horizontal exhibits.

5 2 1 for a pH sensitive polymer coated PEDV vaccines recognizes a difference when, at pH 1 5 1 5 (low ->high) is based on the resulting pH 8 exhibits once again is divided into two difference.

As a result, cationic polymer layer and pH sensitive polymer layer when both formed, regardless of coating layer order (3 and 4 in the case of low ->high at once), stable at low pH under conditions virus vaccine particles were confirmed. Alternatively, coating layer in inactivated, pH sensitive polymer to form only on the basis of identifying capable of reliability and stability.