Compositions and devices for vaccine release and their use

Abstract

The present invention provides a device and method for delivering an effective amount of therapeutic agent, such as a vaccine, to a target that needs it. [Solution] Microneedles and microneedle devices, kits, and methods for manufacturing and using them, including implantable tips (e.g., silk-based tips) for sustained skin delivery of coronavirus and / or influenza vaccines, are described herein. In other embodiments, compositions and methods for controlled or sustained administration of coronavirus and / or influenza vaccines are also described for providing improved immunogenicity and / or broad-spectrum immunity.

Claims

1. A microneedle device (e.g., a microneedle patch) comprising multiple microneedles, A first microneedle comprising a coronavirus antigen (e.g., one or more coronavirus antigens, e.g., SARS-CoV-2 antigen) or a vaccine preparation thereof ("coronavirus vaccine"), Optionally, a second microneedle containing an influenza antigen (e.g., one or more influenza antigens, or a vaccine preparation thereof ("influenza vaccine")) and Includes, A microneedle device configured to deliver coronavirus antigen or coronavirus vaccine, and optionally influenza antigen or influenza vaccine, to a target in an amount sufficient to induce, for example, an immune response (e.g., humoral and / or cellular immune response).

2. In a plurality of microneedles, the first and / or second microneedles are (i) a base (for example, a soluble base) (ii) A tip applied to the base (e.g., a transplantable tip), (iii) Optionally, the backing applied to the base and The microneedle device according to claim 1, including the following:

3. The microneedle device according to claim 1 or 2, wherein the first and / or second microneedles comprise silk fibroin, for example, regenerated silk fibroin and / or recombinant silk fibroin.

4. A microneedle device according to any one of the prior claims, wherein the tip of the microneedle is a silk fibroin tip (for example, an implantable silk fibroin tip).

5. A microneedle device (e.g., a microneedle patch) comprising multiple silk fibroin-based microneedles, Multiple microneedles (e.g., multiple first microneedles) include coronavirus antigens, e.g., one or more coronavirus antigens (e.g., one or more of SARS-CoV-2 antigen, SARS-CoV antigen, or MERS-CoV antigen), or a vaccine preparation thereof ("coronavirus vaccine") A microneedle device is configured to deliver coronavirus antigen or coronavirus vaccine to a target in an amount sufficient to induce, for example, an immune response (e.g., humoral and / or cellular immune response).

6. Microneedles (i) a base (for example, a soluble base) (ii) A silk fibroin tip containing silk fibroin applied to a base (e.g., an implantable silk fibroin tip), (iii) Optionally, the backing applied to the base and The microneedle device according to claim 5, including the following:

7. The microneedle device according to claim 5 or 6, further comprising a second microneedle or a plurality of microneedles containing an influenza antigen, for example, an influenza vaccine.

8. A microneedle device according to any one of the prior claims, wherein the tip of the microneedle contains a coronavirus vaccine or an influenza vaccine.

9. A microneedle device according to any one of claims 1 to 4 or 7, wherein the tip of the microneedle contains a coronavirus vaccine and an influenza vaccine.

10. A microneedle device according to any one of the prior claims, wherein the microneedle base (e.g., a soluble base) comprises a coronavirus vaccine and / or an influenza vaccine.

11. A microneedle device according to any one of the prior claims, wherein the first and / or second microneedles contain one antigen per microneedle, for example, one vaccine per microneedle.

12. The microneedle device according to any one of claims 1 to 4 or 7 to 11, wherein the first microneedle includes a combination of antigens derived from the same coronavirus (e.g., a combination of SARS-CoV-2 antigens) or a combination of antigens derived from different coronaviruses, e.g., different betacoronaviruses.

13. The microneedle device according to any one of claims 1 to 4 or 7 to 12, wherein the second microneedle contains a combination of antigens derived from the same influenza virus or different influenza viruses.

14. The microneedle device according to any one of the prior claims, wherein the device or plurality further comprises a plurality of additional microneedles ("additional plurality"), wherein one or more microneedles in the plurality of additional microneedles (e.g., each microneedle) comprises an influenza vaccine, e.g., one, two, three, four or more influenza vaccines.

15. The microneedle device according to claim 14, wherein the additional multiple comprises one or more microneedles containing, for example, a co-formulated influenza vaccine combined with a coronavirus vaccine, within the same microneedle.

16. The microneedle device according to claim 14 or 15, wherein each additional microneedle contains a coronavirus vaccine and an influenza vaccine, for example, one, two, three, four or more influenza vaccines within the same microneedle.

17. The microneedle device according to claim 14, comprising, for example, two, three, four, or more additional influenza vaccines co-formulated, for example, in the same microneedle, separate from the coronavirus vaccine, wherein multiple (for example) additional microneedles exist in combination.

18. The microneedle device according to claim 14, wherein one or more influenza vaccines and coronavirus vaccines are each formulated separately in individual microneedles.

19. The plurality of microneedles comprising at least two, three, four, five, six, or more antigens (for example, at least five antigens) according to any one of the prior claims Microneedle devices.

20. The microneedle device according to claim 19, wherein some or all of the multiple microneedles are the same.

21. A microneedle device according to any one of claims 1 to 4 or 7 to 20, wherein the plurality of microneedles include coronavirus antigens and influenza antigens, for example, at least one, two, three, four or five, or more influenza antigens, for example, at least four influenza antigens.

22. The microneedle device according to claim 21, wherein some or each of the multiple microneedles include a combination of coronavirus antigen and influenza antigen, for example, at least one, two, three, four or five or more influenza antigens, for example, at least four influenza antigens.

23. The microneedle device according to claim 22, wherein each of the multiple microneedles contains a combination, for example, coronavirus and influenza antigens co-formulated, and for example, all microneedles are the same.

24. The microneedle device according to claim 21, wherein at least two, three, four, or five or more antigens, for example, each antigen is individually formulated, for example, one antigen per microneedle.

25. The microneedle device according to claim 24, comprising at least five different microneedles, each microneedle comprising at least one different antigen.

26. A microneedle device according to any one of claims 1 to 4 or 7 to 25, wherein multiple of the devices comprise at least one coronavirus antigen and at least two, three, four or five or more influenza antigens, for example, at least four influenza antigens.

27. The microneedle device according to claim 26, wherein each antigen is formulated individually.

28. A microneedle device according to any one of the prior claims, wherein the plurality of microneedles comprise at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more (e.g., at least 20%) of the microneedles containing a coronavirus antigen, for example, one or more coronavirus antigens.

29. A microneedle device according to any one of claims 1 to 4 or 7 to 28, wherein the plurality of microneedles comprise at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more (e.g., at least 80%) of the microneedles, each containing an influenza antigen, for example, one, two, three, or four influenza antigens.

30. Multiple microneedles, (i) at least 10% microneedles each containing coronavirus antigens, e.g., one or more coronavirus antigens; and at least 90% microneedles each containing influenza antigens, e.g., one, two, three or four influenza antigens; (ii) at least 20% microneedles each containing coronavirus antigens, e.g., one or more coronavirus antigens; and at least 80% microneedles containing influenza antigens, e.g., one, two, three or four influenza antigens; or (iii) Microneedles containing at least 30% each coronavirus antigen, e.g., one or more coronavirus antigens; and microneedles containing at least 70% each influenza antigen, e.g., one, two, three or four influenza antigens. A microneedle device according to any one of claims 1 to 4 or 7 to 29, including the following:

31. A microneedle device according to any one of the prior claims, wherein the coronavirus vaccine comprises SARS-CoV-2 antigen (e.g., SARS-CoV-2-S1 or its subunits), MERS-CoV antigen (e.g., MERS-CoV-S1 or its subunits), SARS-CoV antigen (e.g., SARS-CoV-S1 or its subunits), or a combination thereof.

32. A microneedle device according to any one of the prior claims, wherein the coronavirus vaccine comprises a SARS-CoV-2 vaccine, for example, a SARS-CoV-2 gene product (e.g., SARS-CoV-2 protein, or nucleic acid encoding the SARS-CoV-2 protein (e.g., mRNA, DNA)), a virus or viral particle (e.g., an inactivated or attenuated SARS-CoV-2 virus), or a viral vector, or a combination thereof.

33. A microneedle device according to any one of the prior claims, wherein the SARS-CoV-2 vaccine is selected from SARS-CoV-2 spike protein or its subunits, SARS-CoV-2 nucleocapsid protein, inactivated virus (e.g., inactivated SARS-CoV-2, e.g., UV-inactivated SARS-CoV-2), viral vector (e.g., non-replicating viral vector or replicating viral vector), virus-like particles, DNA (e.g., DNA plasmid), RNA (e.g., messenger RNA (mRNA), self-amplified mRNA (saRNA), nucleoside-modified mRNA (modRNA), or uridine-containing mRNA (uRNA)), adenovirus vector (e.g., adenovirus type 5 vector, e.g., Ad5-nCoV), or a combination thereof.

34. The coronavirus vaccine contains: coronavirus spike protein (e.g., pre-fusion SARS-CoV-2 spike protein), inactivated SARS-CoV-2 (e.g., UV-inactivated SARS-CoV-2, or PiCoVacc); mRNA (e.g., mRNA encoding the coronavirus protein, e.g., mRNA encoding the SARS-CoV-2 spike protein, or its subunits, e.g., mRNA-1273); and adenovirus vectors (e.g., adenovirus type 5 vector expressing the coronavirus protein (e.g., SARS-CoV-2 spike protein or its subunits)). A microneedle device according to any one of the prior claims, selected from the group consisting of: -, for example, Ad5-nCoV; DNA plasmid (for example, a DNA plasmid encoding the SARS-CoV-2 spike protein or its subunits), for example, INO-4800); dendritic cells (for example, dendritic cells modified with a lentiviral vector to express the SARS-CoV-2 minigene), for example, LV-SMENP-DC); artificial antigen-presenting cells (aAPCs), for example, aAPCs modified with a lentiviral vector to express the SARS-CoV-2 minigene), or a combination thereof.

35. A microneedle device according to any one of the prior claims, wherein the coronavirus vaccine comprises a spike protein, for example, a pre-fusion SARS-CoV-2 spike protein or a subunit thereof.

36. Coronavirus vaccine, (i) the whole spike protein, stabilized spike protein, locked spike protein, spike protein subunit, or a receptor-binding domain (RBD) derived from the spike protein; or (ii) A microneedle device according to any one of the prior claims, comprising a spike protein or its subunits, for example, a whole spike protein, a stabilized spike protein, a locked spike protein, a spike protein subunit, or a vector encoding a receptor-binding domain (RBD) derived from a spike protein (e.g., adenovirus type 5 vector), RNA (e.g., mRNA, saRNA, modRNA, or uRNA), or DNA (e.g., a DNA plasmid).

37. A microneedle device according to any one of the prior claims, wherein the coronavirus vaccine comprises inactivated SARS-CoV-2, for example, UV-inactivated SARS-CoV-2.

38. A microneedle device according to any one of the prior claims, wherein the coronavirus vaccine comprises a recombinant protein, for example, recombinant SARS-CoV-2 spike protein, or a subunit thereof.

39. A microneedle device according to any one of the prior claims, wherein the coronavirus vaccine comprises a coronavirus-derived protein, for example, a pre-fusion SARS-CoV-2 spike protein including a trimer structure.

40. A microneedle device according to any one of the prior claims, wherein the coronavirus vaccine comprises a lipid nanoparticle (LNP) formulation, for example, a coronavirus vaccine encapsulated by LNPs, for example, an mRNA vaccine encapsulated by LNPs.

41. A microneedle device according to any one of the prior claims, wherein the coronavirus vaccine comprises mRNA-1273, BNT162, INO-4800, Ad26 SARS-CoV-2, TNX-1800, or PiCoVacc, or a combination thereof.

42. A microneedle device according to any one of claims 1 to 4 or 7 to 41, wherein the influenza vaccine comprises a monovalent (e.g., single-valent) or polyvalent influenza vaccine (e.g., bivalent, trivalent, tetravalent, or pentavalent influenza vaccine).

43. A microneedle device according to any one of claims 1 to 4 or 7 to 42, configured to deliver (e.g., release) two or more influenza antigens (e.g., three or more, or four or more antigens) to protect, for example, from two or more different influenza viruses.

44. A microneedle device according to any one of claims 1 to 4 or 7 to 43, wherein the plurality of microneedles include a third microneedle containing an influenza antigen different from the influenza antigen in the second microneedle.

45. Claim 44, wherein the plurality of microneedles include a fourth microneedle containing an influenza antigen different from the influenza antigens in the second and third microneedles. The microneedle device described in [reference].

46. The microneedle device according to any one of claims 44 or 45, wherein the plurality of microneedles include a fifth microneedle containing an influenza antigen different from the influenza antigens present in the second, third, and fourth microneedles.

47. (i) The combination of influenza antigens in the second and third microneedles includes a bivalent influenza vaccine; (ii) The combination of influenza antigens in the second, third, and fourth microneedles comprises a trivalent influenza vaccine; and / or (iii) The combination of influenza antigens in the second, third, fourth, and fifth microneedles includes a quadrivalent influenza vaccine. A microneedle device according to any one of claims 44 to 46.

48. A microneedle device according to any one of claims 1 to 4 or 7 to 47, wherein the influenza vaccine comprises an influenza A vaccine, an influenza B vaccine, an influenza C vaccine, and / or an influenza D vaccine.

49. A microneedle device according to any one of claims 1 to 4 or 7 to 48, wherein the influenza vaccine comprises an influenza A vaccine (e.g., an H1N1 vaccine and / or an H3N2 vaccine) and an influenza B vaccine (e.g., a B / Yamagata lineage and / or a B / Victoria lineage vaccine).

50. A microneedle device according to any one of claims 1 to 4 or 7 to 49, wherein multiple microneedles contain an influenza vaccine versus a coronavirus vaccine in a 4:1 ratio.

51. A microneedle device according to any one of claims 1 to 4 or 7 to 50, wherein the combination of coronavirus vaccine and influenza vaccine includes a pentavalent vaccine.

52. A microneedle device according to any one of the prior claims, configured to provide single-dose protection, e.g., immunity, against coronavirus and / or influenza virus.

53. A microneedle device according to any one of claims 1 to 4 or 7 to 52, configured to deliver to a target an amount of influenza vaccine sufficient to induce an immune response, for example, a humoral and / or cellular immune response.

54. A microneedle device according to any one of claims 3 to 53, wherein the silk fibroin comprises recycled silk fibroin and / or recombinant silk fibroin.

55. A microneedle device according to any one of the prior claims, configured for the sustained release of a coronavirus vaccine and / or influenza vaccine.

56. Sustained release involves substantially continuous low-dose administration of the coronavirus vaccine and / or influenza vaccine, for example, the coronavirus vaccine and / or influenza vaccine between about 1 ug and about 500 ug over a period including at least about 4 days (e.g., about 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more, for example, between about 5 and about 25 days, between about 10 and about 20 days, between about 10 and about 15 days, between about 12 and about 16 days, or between about 14 and about 15 days). The microneedle device according to claim 55.

57. A microneedle device according to any one of the prior claims, wherein the microneedle is configured to release a coronavirus vaccine and / or influenza vaccine to the target skin over a period of time including at least about four days (for example, about four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen days or more, for example, between about five and about twenty-five days, between about ten and about twenty-two days, between about ten and about fifteen days, between about twelve and about sixteen days, or between about fourteen and about fifteen days).

58. A microneedle device according to any one of the prior claims, wherein the microneedle comprises at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25% or more of a dose of the coronavirus vaccine, for example, a standard dose.

59. A microneedle device according to any one of the prior claims, wherein the microneedle tip (e.g., silk fibroin tip) contains silk fibroin at a concentration of about 1% w / v to about 10% w / v (e.g., silk fibroin having a molecular weight distribution of about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10% w / v as shown in Figure 4), or contains, for example, an amount of silk fibroin between about 20 μg and about 245 μg per 121 microneedle array.

60. The base (for example, a soluble base) (i) Polysaccharides (e.g., dextran); (ii) Disaccharides (e.g., sucrose, maltose, and trehalose); (iii) Polymers (e.g., methylcellulose, polyethylene glycol (PEG), carboxymethylcellulose (CMC), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), and hyaluronate); (iv) Proteins (e.g., gelatin); (v) Plasticizers (e.g., glycerol, propanediol); and (vi) Surface-active substances (e.g., octylphenol ethoxylate (e.g., Triton-X), polysorbate, poloxamer, and / or polyethoxylated alcohols) A microneedle device according to any one of the prior claims, comprising two or more of the above.

61. A microneedle device according to any one of the prior claims, wherein the base comprises one or more of the following: gelatin, dextran, glycerol, polyethylene glycol (PEG) (e.g., including low molecular weight PEG), sucrose, trehalose, maltose, carboxymethylcellulose (CMC), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), hyaluronate, methylcellulose, and / or surfactants (e.g., octylphenol ethoxylate (e.g., Triton-X), polysorbate, poloxamer, e.g., P188, and / or polyethoxylated alcohol), and optionally, the microneedles are configured for sustained release.

62. Designed for sustained release, (i) between approximately 20% and approximately 40%, for example, 30% of 70 kDa dextran; (ii) Between approximately 5% and approximately 15%, for example, approximately 10% sucrose; (iii) Glycerol in an amount between approximately 0.5% and approximately 2.5%, for example, about 1%; and (iv) Between approximately 0.001% and approximately 1%, for example, approximately 0.01% of Triton-X; ( (Optionally, this is a solution used for casting and / or for the composition of a dry-solidification base.) A microneedle device according to any one of the prior claims, including the following:

63. The tip of the microneedle (for example, the tip of the silk fibroin) (i) Disaccharides (e.g., sucrose, maltose, and trehalose); (ii) Polymers (e.g., methylcellulose, polyethylene glycol (PEG), carboxymethylcellulose (CMC), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), hyaluronate); (iii) Amino acids (e.g., threonine); (iv) Plasticizers (e.g., glycerol, propanediol); (v) Buffer solution (e.g., PBS); (vi) Surface-active substances (e.g., octylphenol ethoxylate (e.g., Triton-X), polysorbate (e.g., Tween 20), poloxamer, and / or polyethoxylated alcohols); and / or (vii) Adjuvant A microneedle device according to any one of the prior claims, including the following:

64. A microneedle device according to any one of the prior claims, wherein the microneedle (e.g., the tip of the microneedle) is configured to further include and / or release a non-vaccine molecule, e.g., a molecule useful for confirming dose delivery, e.g., a dye molecule (e.g., a biocompatible dye molecule) or a reporter molecule that can be visualized (e.g., by illumination with UV irradiation).

65. A method for providing immunity, e.g., broad-spectrum immunity, to a subject against a virus (e.g., SARS-CoV-2, SARS-CoV, MERS-CoV, and / or influenza), comprising the step of bringing the skin of the subject into contact with a microneedle device described in any one of the prior claims.

66. A method for providing controlled or sustained release of an antigen, for example, a coronavirus vaccine and / or influenza vaccine (e.g., SARS-CoV-2 antigen, SARS-CoV antigen, MERS-CoV antigen, and / or influenza antigen, or a vaccine preparation thereof) to a subject, comprising the step of bringing the skin of the subject into contact with a microneedle device according to any one of claims 1 to 64.

67. A method for enhancing an immune response to a virus (e.g., coronavirus (e.g., SARS-CoV-2, SARS-CoV, and / or MERS-CoV) and / or influenza virus) in a subject, comprising the step of bringing the skin of the subject into contact with a microneedle device according to any one of claims 1 to 64.

68. The immune response (i) Elevated coronavirus-specific antibody titers detectable in the blood of the subject, for example, at least 3, 4, 5, or 6 days after immunization, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, and / or 16 weeks (e.g., SARS-CoV-2 specific antibodies, e.g., antibodies specific to the SARS-CoV-2 spike protein or its subunits); (ii) Elevated anti-coronavirus IgG (e.g., anti-SARS-CoV-2 IgG) titer detectable in the blood of the subject, for example, at least 4, 5, or 6 days after immunization, or at least 1, 2, 3, or 4 weeks, or at least 1, 2, 3, 4, 5, and / or 6 months; (iii) Elevated coronavirus-specific antibody titers detectable in the subject's blood over the entire post-immunization coronavirus season (e.g., SARS-CoV-2 specific antibodies, e.g., antibodies specific to the SARS-CoV-2 spike protein or its subunits); (iv) For example, an increase in the level of IFNγ secreting cells in the subject's blood at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and / or 12 weeks after immunization; and / or (v) For example, increased production of IFNγ per pre-selected number of cells in the subject's blood at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and / or 12 weeks after immunization. The method according to claim 67, wherein the response is a humoral and / or cellular immune response comprising the above.

69. A method for providing broad-spectrum immunity to coronaviruses (e.g., SARS-CoV-2, SARS-CoV, and / or MERS-CoV) and / or influenza viruses in a subject, comprising the step of bringing the skin of the subject into contact with a microneedle device according to any one of claims 1 to 64, for example, to produce an immune response (e.g., a cellular immune response and / or a humoral immune response) in the subject to drift strains of coronaviruses and / or influenza viruses.

70. (i) The coronavirus vaccine contains antigens associated with a first coronavirus strain, and administration of a certain dose of the coronavirus vaccine to a subject results in broad-spectrum immunity against a second coronavirus strain (e.g., drift SARS-CoV-2 strain) that is not present in or associated with the composition or vaccine; (ii) The influenza vaccine contains a first influenza strain, and administration of a certain dose of the first influenza strain to a subject results in broad-spectrum immunity against a second influenza strain (e.g., a drift influenza strain) that is not present in the composition or vaccine; (ii) The influenza vaccine contains a first influenza A strain, and administration of a certain dose of the first influenza A strain to a subject results in broad-spectrum immunity to drift influenza strains (e.g., drift influenza A, B, C, and / or D strains) that are not present in the composition or in the vaccine; (iii) The influenza vaccine contains a first influenza B strain, and administration of a certain dose of the first influenza B strain to a subject results in broad-spectrum immunity against drift influenza strains (e.g., drift influenza A, B, C, and / or D strains) that are not present in the composition or in the vaccine; (iv) The influenza vaccine contains a first influenza C strain, and administration of a certain dose of the first influenza C strain to a subject results in broad-spectrum immunity against drift influenza strains (e.g., drift influenza A, B, C, and / or D strains) that are not present in the composition or in the vaccine; and / or (v) The influenza vaccine contains a first influenza D strain, and administration of a certain dose of the first influenza D strain to a subject results in broad-spectrum immunity against drift influenza strains (e.g., drift influenza A, B, C, and / or D strains) that are not present in the composition or in the vaccine. The method according to any one of claims 65 to 69.

71. Claim 6: An elevated coronavirus-specific antibody titer (e.g., a SARS-CoV-2 specific antibody, e.g., an antibody specific to the SARS-CoV-2 spike protein or its subunits) is detectable in the subject's blood throughout the entire coronavirus season after immunization, and optionally, the elevated coronavirus-specific antibody titer is against a drift coronavirus strain (e.g., a drift SARS-CoV-2 strain). The method described in any one of items 5 to 70.

72. The method according to any one of claims 65 to 71, wherein the seroconversion rate in the blood of the subject is greater than approximately 20% (e.g., 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% or more, e.g., 100%), based on, for example, an elevated coronavirus-specific antibody titer (e.g., SARS-CoV-2 specific antibody, e.g., an antibody specific to the SARS-CoV-2 spike protein or its subunits), detectable in the blood of the subject six months after immunization.

73. The method according to any one of claims 68, 71, or 72, wherein the elevated titer of coronavirus-specific antibodies (e.g., SARS-CoV-2-specific antibodies, e.g., antibodies specific to the SARS-CoV-2 spike protein or its subunits), the elevated titer of anti-coronavirus IgG (e.g., anti-SARS-CoV-2 IgG), and / or the level of antibody-secreting plasma cells (ASCs) against coronavirus (e.g., SARS-CoV-2 virus) is higher (e.g., 1x, 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, 10x, 11x, 12x, 13x, 14x, or 15x or more) compared to a single dose or bolus dose of coronavirus vaccine (e.g., SARS-CoV-2 vaccine).

74. Immune response and / or broad-spectrum immunity, (i) an increase in hemagglutination inhibition (HAI) antibody titer detectable in the blood of the subject, for example, at least 3, 4, 5, or 6 days after immunization, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 51, and / or 52 weeks or longer, which optionally is against drift influenza A, B, C, and / or D strains; (ii) an increase in anti-influenza IgG titer detectable in the blood of the subject, for example, for at least 4, 5, or 6 days after immunization, or at least 1, 2, 3, or 4 weeks, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and / or 12 months or longer, which optionally is against drift influenza A, B, C, and / or D strains; and / or (iii) Levels of antibody-secreting plasma cells (ASCs) against influenza virus, for example, against drift influenza A, B, C, and / or D strains, detectable in the bone marrow of the subject, for example, detectable for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 51, and / or 52 weeks or more after immunization. The method according to any one of claims 65 to 73, comprising a cellular and / or humoral immune response.

75. A method for providing an immune response (e.g., a cellular immune response and / or a humoral immune response) and / or broad-spectrum immunity to coronaviruses (e.g., SARS-CoV-2, SARS-CoV, and / or MERS-CoV) and / or influenza viruses in a subject, comprising the step of bringing the skin of the subject into contact with a microneedle device according to any one of claims 1 to 64, Coronavirus vaccines and / or influenza vaccines are, in the case of A method of administering a drug in an amount (e.g., a dose) sufficient to induce an immune response (e.g., a cellular immune response and / or a humoral immune response) against the ronavirus and / or influenza virus, and / or over a period of about 5 to 25 days (e.g., about 10 to 20 days, about 10 to 15 days, about 12 to 18 days, or about 14 to 15 days), for example, about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 days.

76. The method according to any one of claims 65 to 75, wherein the subject (for example, a human subject) is a child.

77. The method according to any one of claims 65 to 75, wherein the subject (for example, a human subject) is an adult subject.

78. The method according to any one of claims 65 to 75, wherein the subject (for example, a human subject) is an elderly subject.

79. The method according to any one of claims 65 to 78, wherein a coronavirus vaccine and / or an influenza vaccine is administered prophylactically.

80. The method according to any one of claims 65 to 79, wherein a single dose of a coronavirus vaccine and / or influenza vaccine provides, in a subject, protection against, for example, infection by a coronavirus and / or influenza virus.

81. The method according to any one of claims 65 to 80, wherein the coronavirus vaccine and / or influenza vaccine is administered as a patch, for example, a single patch.

82. The method according to claim 81, wherein the patch is administered to the subject by a medical professional (e.g., a doctor or nurse).

83. The method according to claim 81, wherein the patch is self-administered.

84. The method according to any one of claims 81 to 83, wherein the patch is administered using an applicator.

85. The method according to any one of claims 65 to 84, further comprising the release (e.g., controlled or sustained release) of a non-vaccine molecule in the target, such as a molecule useful for confirming dose delivery, such as a dye molecule (e.g., a biocompatible dye molecule) or a reporter molecule that can be visualized (e.g., by illumination with UV irradiation).

86. A method for producing microneedle devices, The steps include providing a mold that includes a mold body having an array of needle cavities of a predetermined shape, for example, pyramidal and / or conical needle cavities formed inside; The steps include filling the tip of a needle cavity with a composition (e.g., a composition comprising silk fibroin), a coronavirus antigen, for example, a coronavirus vaccine (e.g., one or more of SARS-CoV-2 antigen, SARS-CoV antigen, or MERS-CoV antigen, or vaccine preparations thereof), and / or an influenza antigen, for example, an influenza vaccine (e.g., one or more influenza antigens, or vaccine preparations thereof); The filled tip of the needle cavity is dried to create a microneedle tip (e.g., a silk fibroin tip), and the microneedle tip is optionally annealed. Steps to take; The steps include: filling the needle cavity of the mold with a base (e.g., a soluble base) solution; The steps include: drying the base solution to create a base layer for the tip of the microneedle (e.g., the tip of the silk fibroin); The steps include: applying a backing to the base layer as an option to create a microneedle device; and Methods that include...

87. The method according to claim 86, further comprising the step of removing the microneedle device from the mold before optionally applying a backing.

88. The method according to claim 86 or 87, wherein the microneedle device is removed by bending the mold away from the microneedle device.

89. The method according to any one of claims 86 to 88, further comprising the step of packaging a microneedle device in a container with low water vapor transmission with a desiccant that maintains a relative humidity in the package between approximately 0% and approximately 50% (for example, between approximately 0% and 10%, between approximately 10% and approximately 20%, between approximately 20% and approximately 30%, between approximately 30% and approximately 40%, or between approximately 40% and 50%, for example, approximately 25%).

90. The method according to any one of claims 86 to 89, wherein a composition (e.g., a composition comprising silk fibroin), a coronavirus vaccine, and / or an influenza vaccine solution are dispensed into each needle cavity in a mold via nanoliter printing.

91. The method according to any one of claims 86 to 90, wherein the step of filling the tip of the needle cavity includes dispensing a solution, for example, silk fibroin, coronavirus vaccine, and / or influenza vaccine solution, into each needle cavity.

92. A microneedle, microneedle device, or method according to any one of the prior claims, wherein a coronavirus vaccine and / or influenza vaccine is adjuvanted.

93. A microneedle, microneedle device, or method according to any one of the prior claims, wherein the coronavirus vaccine and / or influenza vaccine are not adjuvated.

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