Measles and rubella vaccine delivered by HD-map

Abstract

The present invention relates to stable vaccine formulations for administration via a microprojection array in which the microprojections are densely packed and in which the vaccine formulations are rapidly sprayed or layered on to the microprojections in relatively small amounts such that the formulations dry rapidly. The present invention relates in particular to stabilized measles and rubella vaccines delivered by high density micro-array MAPs (HDMAP).

Description

MEASLES AND RUBELLA VACCINE DELIVERED BY HD-MAPCross-Reference to Related Application

[0001] This application claims priority to US Provisional Patent Application No. 63 / 674,461 filed on 23 July 2024.Background of the Invention

[0002] The present invention relates to stable vaccine formulations for administration via a microprojection array in which the microprojections are densely packed and in which the vaccine formulations are rapidly sprayed or layered on to the microprojections in relatively small amounts such that the formulations dry rapidly. The present invention relates in particular to stabilized measles and rubella vaccines delivered by high density micro-array MAPs (HD- MAP).Description of the Prior Art

[0003] In recent years, attempts have been made to devise new methods of delivering drugs and other bioactive materials, for vaccination and other purposes, which provide alternatives that are more convenient and / or enhanced in performance to the customary routes of administration such as intramuscular and intradermal injection. Limitations of intradermal injection include: cross -contamination through needle-stick injuries in health workers; injection phobia from a needle and syringe; and most importantly, as a result of its comparatively large scale and method of administration, the needle and syringe cannot target key cells in the outer skin layers. This is a serious limitation to many existing and emerging strategies for the prevention, treatment and monitoring of a range of untreatable diseases. There is also a need to reduce the amount of material delivered due to toxicity of the material or due to the need to conserve the material because it is difficult or expensive to produce.

[0004] In an effort to solve some of the issues referenced above microprojection arrays or microneedle arrays have been utilized to deliver various materials through the skin. For example, WO 2005 / 072630 describes devices for delivering bioactive materials and other stimuli to living cells. The devices comprise a plurality of projections which can penetrate theskin so as to deliver a bioactive material or stimulus to a predetermined site. The projections can be solid and the delivery end of the projection is designed such that it can be inserted into targeted cells or specific sites on the skin.

[0005] One of the challenges of using devices that contain microneedles and / or microprojections is the need to coat the projections. Various coating techniques such as dipping the array into a coating solution or spraying the coating onto the projections have been described. For example, Gill and Prausnitz, J. Controlled Release (2007), 117: 227-237 describe coating microprojections by dipping the microprojections into a coating solution reservoir through dip holes that are spaced in accordance with the microprojection array. Cormier et al., J. Controlled Release (2004), 97: 503-511 describe coating microneedle arrays by partial immersion in an aqueous solution containing active compounds and polysorbate. WO 2009 / 079712 describes methods for coating microprojection arrays by spray coating the microprojections and drying the sprayed solution with gas.

[0006] Rapid spray coating of microprojection / microneedle drug delivery and vaccine platforms allow allocation of the coating to the delivery platform minimizing the inefficiencies associated with spray coating or dip coating that may overcoat or undercoat the microprojections. Moreover, dip coating or spray coating is less accurate than inkjet coating. Many vaccines are comprised of multiple valencies that may be for protection against a single pathogen such as a thirteen valent vaccine against pneumococcal infections or multiple pathogens such as MMR vaccine against measles mumps and rubella. Such vaccines containing more than one factor may have incompatibilities among the various factors or among the various excipients or solvents used to deliver the vaccine or to make the vaccine more efficacious. Moreover, designing a stable vaccine with multiple valencies that may be distributed on a surface such as a microneedle or microprojection and dried poses challenges. In addition, each component of the multivalent vaccine composition affects the viscosity, drop formation, dry time, adhesion and stability of the vaccine. Other challenges to delivering a complex vaccine via a microprojection / microneedle array include coating the microneedles / microprojections with enough vaccine to be efficacious when administered, formulating a vaccine such that the drop size is sufficiently small to permit penetration into the skin with each projection of the array. There is also a need to providemicroneedle / microprojection arrays that enable coating of the microneedle / microproj ection with compositions that have components that are incompatible with each other in solution. In other words, it may be desirable to have microneedle / microprojection arrays that can be coated by a device such that each of the components to be delivered is separately coated on to the microneedle / microprojections.

[0007] Although there are clear benefits with combination vaccines, the main challenge in their development is the risk that the efficacy or safety of the combination would be less than that seen with the administration of the vaccines separately. New combinations cannot be less immunogenic, less efficacious, or more reactogenic than the previously licensed uncombined vaccines. Immunological, physical, and / or chemical interactions between the combined components have the potential to alter the immune response to specific components. Finally, and ideally, the many advantages of combination vaccines should not be achieved at the cost of reduced product stability. From a practical standpoint, uncommon transport and storage conditions and could hamper the development of a combination vaccine. The present invention provides compositions and methods for making stable measles and rubella vaccines that may be coated onto a HD-MAP and delivered via a device for immunizations.

[0008] The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that the prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.Summary of the Present Invention

[0009] The present invention relates to devices and methods for coating microprojections on microneedle arrays with various substances. These substances may be liquid or non-liquid and may be coated onto the microprojection array by mjet spraying. The present invention also relates to microprojection arrays having a base and a plurality of microprojections.

[0010] The present invention also relates to devices, formulations and methods for coating vaccines onto microprojections of a microprojection array such that the vaccines are more stable than corresponding vaccines is solution. The present invention provides increasedstability of vaccine formulations based on antigen potency as measured by various methods including ELISA before and after rapid drying.

[0011] The present invention provides increased stability of vaccine formulations based on antigen potency as measured by various methods including ELISA after drying and storage at various temperatures such 2-8°C and 25°C and elevated temperatures such as 45°C.

[0012] Increase stability of vaccine formulations based on antigen potency as measured by various methods including ELISA due to the use of particular excipients at particular percentage ranges in combination with rapid drying due to the use of a rapid jetting device.

[0013] In one broad form an aspect of the present invention relates to vaccine compositions for coating a microprojection array having two or more microprojections comprising measles virus and rubella virus and sorbitol, L-histidine, trehalose dihydrate sodium phosphate dibasic dihydrate, sodium phosphate monobasic dihydrate and hydrolyzed porcine gelatin.

[0014] In one embodiment the present invention relates to vaccine compositions containing 0.5 to 5% w / v sorbitol, lOmM to lOOmM L-histidine, 0.5 to 4% w / v trehalose dihydrate w / v, ImM to lOmM sodium phosphate dibasic dihydrate, ImM to lOmM sodium phosphate monobasic dihydrate and 0.5 to 4% w / v hydrolyzed porcine gelatin.

[0015] In one embodiment the present invention relates to vaccine compositions containing 2% w / v sorbitol, 50mM L-histidine, 1% w / v trehalose dihydrate w / v, 5.4mM sodium phosphate dibasic dihydrate, 4.6mM sodium phosphate monobasic dihydrate and 1% w / v hydrolyzed porcine gelatin.

[0016] In one embodiment the present invention relates to vaccine compositions where the pH is between 6.5 and 7.0.

[0017] In one embodiment the present invention relates to vaccine compositions where the amount of rubella and measles virus is at least 1000 CCID50 for each virus and where rubella virus is between 3000 to 5000 CCID50 and measles virus is 2000 to 4000 CCID50.

[0018] In one embodiment the present invention relates to compositions where the formulation is sprayed onto the microprojections of the microprojection array and the composition dries on the microprojections.

[0019] In one embodiment the present invention relates to compositions which are sprayed onto the microprojection arrays where the microprojection array comprises a base and solid microprojections which project from the base and wherein the microprojections are 300 to 400pm in length and the array is made of a polymer including but not limited to liquid crystal polymers,

[0020] In one embodiment the present invention relates to microprojection arrays where the projections extending from the base wherein the projections comprise a targeting section for delivering the viruses to thereby cause an immunological response and a cylindrical support section for supporting the targeting section, wherein two or more steps exist between the targeting section and the support section and the support section is wider than the targeting section.

[0021] In one embodiment the present invention relates to composition of claim 7 wherein the composition is stable for at least 3 or 6 or 12 or 24 or 36 months.

[0022] In one embodiment the present invention relates to compositions where the stability is measured by titer or by genomic material.

[0023] In one embodiment the present invention relates to methods of immunizing a human against measles and rubella by administering to the human the measles-rubella composition by projecting the microprojection array into the human’s skin thereby penetrating the skin with the microprojections coated with the composition.

[0024] In one embodiment the present invention relates to methods of immunization where the seroconversion rate for neutralizing antibodies with respect to measles is between 10 to 50% for subjects that were previous exposed to measles either by contracting the disease or by previous vaccination and the seroconversion rate for neutralizing antibodies with respect to rubella is between 10 to 50% for subjects that were previous exposed to rubella either by contracting the disease or by previous vaccination.

[0025] Array as used herein refers to devices that include one or more structures such as microprojections capable of piercing the stratum corneum to facilitate transdermal delivery of therapeutic agents through or to the skin.

[0026] Microprojections, as used herein, refers to the specific microscopic structures associate with the array that are capable of piercing the stratum corneum to facilitate transdermal delivery of therapeutic agents through or to the skin. Microprojections may include needle or needlelike structures, micro-pins as well as solid projections.

[0027] It will be appreciated that the broad forms of the invention and their respective features can be used in conjunction, interchangeably and / or independently, and reference to separate broad forms is not intended to be limiting.Brief Description of the Drawings

[0028] Various examples and embodiments of the present invention will now be described with reference to the accompanying drawings, in which: -

[0029] Figure 1 A is a plot of long-term thermostability (potency) at 2-8 °C for measles on MR HD-MAP used for clinical study; Figure IB is a bar chart of stability at all temperatures at 0, 12 and 24 months for measles on MR HD-MAP; Figure 1C is a plot of long-term thermostability (potency) at 2-8 °C for rubella on MR HD-MAP used for clinical study; Figure ID is a bar chart of stability at all temperatures at 0, 12 and 24 months for rubella on MR HD- MAP.

[0030] Figure 2 A is a plot of neutralizing antibody concentrations for measles. Figure 2B is a plot of neutralizing antibody concentrations for rubella. Serum was collected from subjects at Day 0, 7, 28 and 56 and tested for neutralizing antibodies in the FRNT50 assay, normalized to WHO IS sera run in parallel. The geometric mean of the FRNT50 value (lU / mL) and 95% CI are shown for each group and day. The dotted line on the y-axis of each graph represents the protective threshold for each virus (0.120 lU / mL for measles, 10 lU / mL for rubella).

[0031] Figure 3A is a plot for measles FRNT50 DO to D28 Fold Change. FRNT50 fold change between DO and D28 for all groups; bar represents geometric mean. Figure 3B is a plot of therelationship between DO titre (x axis) and D28 fold change (y axis) is shown. In both graphs, each point represents a single subject, coloured by group. Dotted lines represent seroconversion (fold change > 4) or no change (fold change = 1).Detailed Description of the Preferred Embodiments

[0032] The present invention relates to devices and methods for coating microprojections on microneedle arrays with various substances. These substances may be liquid or non-liquid and may be coated onto the microprojection array by mjet spraying. The present invention also relates to microprojection arrays having a base and a plurality of microprojections.

[0033] Microprojection and microneedle arrays can be in the form of MAP having projections extending from a surface of a base. The projections and base may be formed from any suitable material, including but not limited to silicon and various polymers including liquid crystal polymers. The projections may be solid, non-porous and non-hollow.

[0034] In an array the MAP has a width W and a breadth B with the projections being separated by spacing. The projections may be provided in an array that is defined by a regular iteration of microprojections along a square or rectangular arrangement, but other arrangements of projections such as circular arrangement of the projections that are compatible with rotational spray coating may also be used. In order to further improve or enhance the targeting accuracy, the substrate may be designed such that the features to be coated are located on radial lines from the center point of the rotation or located on concentric circles or on a continuous spiral. The substrate may be designed such that the feature spacing on each arc is designed to match an integer number of steps of the motor for a given radius. Each projection includes a tip for penetrating tissue of the biological subject and projections will typically have a profile which tapers from the base to the tip.

[0035] The MAP is applied to the biological subject by positioning the MAP against a surface of a subject or by positioning the MAP near the subject if an applicator that can propel the MAP toward the skin is utilized. The tips of the projections penetrate the surface of the skin and may penetrate tissue beneath the surface of the skin to a given depth as the MAP is applied. The MAP may be used to deliver material or stimulus to internal tissues of a patient. The MAP may be delivered such that the projections pierce the Stratum Comeum (SC) and penetratethrough the Viable Epidermis VE to penetrate the Dermis DE) by a dermal penetration depth. The MAP may be used to deliver material or stimulus to any part or region in the subject. The MAP can be provided in a variety of different configurations to suit different material or stimulus delivery requirements. Accordingly, the specific configuration of the MAP can be selected to allow the delivery of material and stimulus to particular tissues, at a specific depth, to induce a desired response.

[0036] The microprojection arrays that the applicator of the present invention projects into the skin may have a variety of shapes and sizes. The microprojection array may be square, circular, rectangular or irregular depending on its use. In some embodiments the microprojection arrays are square and have an equal number of microprojections in each row. For example the microprojection array may have 30 rows of 30 microprojections for a 30 x 30 array of 900 microprojections or 40 rows of 40 microprojections for a 40 x 40 array of 1600 microprojections or 50 rows of 50 microprojections for a 50 x 50 array of 2500 microprojections or 60 rows of 60 microprojections for a 60 x 60 array of 3600 or 70 rows of 70 microprojections for a 70 x 70 array of 4900 microprojections. The microprojection arrays may be in the shape of a rectangle where the number of rows does not equal the number of microprojections in a row. For example the microprojection array may have 30 rows of 40 microprojections for a 30 x 40 array of 1200 microprojections or 40 rows of 50 microprojections for a 40 x 50 array of 2000 microprojections or 50 rows of 60 microprojections for a 50 x 60 array of 3000 microprojections.

[0037] The microprojection arrays may be divided into areas such that a different vaccine antigen or other substance such as an excipient may be coated in each area. For example, the microprojection array may be divided in half or into four equal quadrants where different vaccine antigens or other substances such as excipients may be applied. These areas may have equal numbers of microprojections or unequal numbers of microprojections. In other embodiments some of the microprojections may be uncoated.

[0038] The microprojection arrays can be varied in size depending on its use. The area of the MAP will have an impact on the ability to penetrate the subject, but this must be balanced by the need to induce cell damage over a sufficiently large area to induce a response. Consequentlythe MAPs typically have dimensions of between 0.5 x 0.5 mm and 20 x 20 mm, between 0.5 x 0.5 mm and 15 x 15 mm and more typically between 1 x 1 mm and 12 x 12 mm.

[0039] In one embodiment the microprojection array is 10.6x10.6mm. The microprojection arrays may have a density of projections of between 1,000 to 20,000 per cm2or from 1,000 to 15,000 per cm2, or from 1,000 to 10,000 per cm2for from 1,000 to 5,000 per cm2, or from 2,500 to 20,000 per cm2or from 2,500 to 15,000 per cm2or from 2,500 to 10,000 per cm2or from 2,500 to 7,500 per cm2or from 2,500 to 5,000 per cm2or from 5,000 to 20,000 per cm2or from 5,000 to 15,000 per cm2or from 5,000 to 10,000 per cm2or from 5,000 to 9,000 per cm2or from 5,000 to 8,000 per cm2or from 5,000 to 7,000 per cm2or from 5,000 to 6,000 per cm2. The applicators of the present invention are often utilized to project high density microprojection arrays into the skin. Such high density arrays are microprojection arrays of sufficient size and density such that forces that can be applied manually will be insufficient to overcome the elasticity of the skin. The projections are typically separated by between 10 pm and 200 pm, between 30 pm and 150 pm, between 50 pm and 120 pm and more typically between 70 pm and 100 pm, leading to MAPs having between 10 and 1000 projections per mm2and more typically between 100 and 3000 projections per mm2, and in one specific example approximately 20,000 per cm2.

[0040] The length of the projections may be from 100pm to 500pm or from 100pm to 400pm or from 100pm to 300pm or from 100pm to 250pm or from 100pm to 200pm or from 150pm to 500pm or from 150pm to 400pm or from 150pm to 300pm or from 150pm to 250pm or from 150pm to 200pm or from 200pm to 700pm or from 200pm to 600pm or from 200pm to 500pm or from 200pm to 400pm or from 200pm to 300pm or from 200pm to 250pm or from 225pm to 700pm or from 225pm to 600pm or from 225pm to 500pm or from 225pm to 400pm or from 225pm to 300pm or from 225pm to 250pm or from 250pm to 700pm or from 250pm to 600pm or from 250pm to 500pm or from 250pm to 400pm or from 250pm to 300 pm. In one embodiment of the HD-MAP the length of the microprojections is 350pm.

[0041] The projections may have a step shoulder (discontinuity) between the cone and pillar of the projection. In the event that a discontinuity is provided, this is typically located so that as the discontinuity reaches the dermis, penetration of the projection stops, with the tip extending into the dermal layer. Typically the discontinuity is located from the end of the tipat between 10 and 100 m, between 20 and 90 pm, between 30 and 80 pm, and more typically between 40 and 60 pm.

[0042] The microprojection array may be made of any suitable materials including but not limited to silicon, polymers, and plastic. In silicon embodiments the base thickness is about 60 um or silicon with a thin (1mm) polymer backing. The overall mass of some embodiments of the microprojection array is about 0.8 gm. The microprojection array may have bevelled edges to reduce peak stresses on the edge of the array. The MAP can be quartered or subdivided by other ratios to reduce the stress load on the MAP and mitigate MAP breakage. Polymer embodiments may have reduced mass. The microprojection array may also have an overall weakly convex shape of the MAP to improve the mechanical engagement with skin and mitigate the effect of high speed rippling application: a ‘high velocity / low mass’ system. The microprojection array may have a mass of less than 1 gram, or less than 0.9 grams or less than 0.8 grams or less than 0.7 grams, or less than 0.6 grams or less than 0.5 grams or less than 0.4 grams. The microprojection array may have a mass of from about 0.1 grams to about 1.0 grams or from about 0.1 grams to about 0.9 grams, or from about 0.1 grams to about 0.8 grams or from about 0.1 grams to about 0.7 grams, or from about 0.1 grams to about 0.6 grams or from about 0.1 grams to about 0.5 grams or from about 0.1 grams to about 0.4 grams, or from about 0.1 grams to about 0.3 grams or from about 0.1 grams to about 0.2 grams. In one embodiment of the applicator / microprojection system the mass of the array is about 0.3 grams, the array is projected at a velocity of about 20-26 m / s by the applicator.

[0043] The projection spacing is selected so that material from the projections is able to at least partially, and optionally at least completely, diffuse between the projections. Accordingly, the projections are typically separated by between 10 pm and 200 pm or between 10 pm and 190 pm or between 10 pm and 180 pm or between 10 pm and 170 pm or between 10 pm and 160 pm or between 10 pm and 150 or between 10 pm and 140 pm or between 10 pm and 130 pm or between 10 pm and 120 pm or between 10 pm and 110 pm or between 10 pm and 100 pm or between 10 pm and 90 pm or between 10 pm and 80 pm or between 10 pm and 70 pm or between 10 pm and 60 pm or between 10 pm and 50 pm or between 10 pm and 40 pm or between 10 pm and 30 pm or between 10 pm and 20 pm

[0044] In some embodiments, more than one coating may be applied to the same projection. For instance, different coatings may be applied in one or more layers to provide the same or different materials for delivery to the tissues within the subject at the same time or different times if the layers dissolve in sequence. A first coating may be applied to modify surface properties of the projection and improve the ability of the second coating to coat the projection in a desirable manner. Multiple layers of the same coating formulation may be used with drying between each layer to allow a progressive build-up of coating to achieve a specific thickness and thus modify the effective cross section of the projection even further.

[0045] Substances applied to the microprojections can be of various types including but not limited to small chemical or biochemical compounds including antigens, ligands, drugs, metabolites, amino acids, sugars, lipids, saponins, and hormones; macromolecules such as complex carbohydrates, phospholipids, peptides, polypeptides, proteins, peptidomimetics, and nucleic acids; or other organic (carbon containing) or inorganic molecules; and particulate matter including whole cells, bacteria, viruses, virus-like particles, cell membranes, dendrimers and liposomes or combinations thereof. Substances may also include contrast enhancing reagents or surface modifying materials.

[0046] The substances may be comprised of a single compound or multiple compounds. For example, in embodiments used for vaccination the microprojections may be coated with a vaccine compound that contains a single antigen or multiple antigens either to the same pathogen or to different pathogens. In another embodiment the substance may be a vaccine composition having an excipient and one or more antigens. In another embodiment the substance may be a vaccine composition having an adjuvant and one or more antigens. As described above vaccine compositions may be delivered by the MAP such that different antigens are located on different microprojections either independent one from another or in sections located on the MAP.

[0047] Coatings may be liquid or non-liquid. Liquid coating materials may aqueous, however other coating solutions are possible, and that the surface properties of the projection may need to be modified to accommodate a range of coating solutions. For an aqueous coating solution, the microprojections may be modified to be more "hydrophobic" in nature. A hydrophilic surface will cause an aqueous solution to completely wet it (assuming low viscosity). Thiswould result in a large fraction of the liquid coating material being wicked onto the base of the projection array, which would impede its delivery to the skin. Increasing the solution viscosity slows down the wicking (or surface wetting) process. If a dry coating process is accomplished rapidly in comparison to the surface wetting, a larger fraction of the liquid coating material can be localized to the projections. By changing the contact angle of the projection surface (by chemically modifying it), the liquid coating solution wetting properties may also be altered. In making the surface more "hydrophobic", an aqueous coating solution will be inhibited from wetting the projection surface down to the base. Furthermore, a surfactant can be added to an aqueous coating solution which is placed on a "hydrophobic" projection. The surfactant may assist in wetting the hydrophobic surface by orienting the polar and non-polar groups of the surfactant at the surface, thus facilitating the wetting. If appropriate drying conditions (either with or without surfactant) are achieved, the result is that a significant portion of the coating material is retained near the projection tips. Striking a balance between the surface wetting properties (i.e. contact angle), solution viscosity, and the presence or absence of a surfactant (among other solution properties) can change the degree and uniformity with which the coating solution is localized to the projection tips. In a further embodiment, the microprojection surface may be altered such that the tips are hydrophilic and the lower portion of the shaft and base are hydrophobic. This can be accomplished using bulk lithographic processes. In this embodiment, the hydrophilic tip surface is easily wet, while the lower portion of the projection inhibits liquid travel towards the base due to its hydrophobic nature. Other methods of coating the microprojections include but are not limited to differential coatings using plasma polymers, spin coating, microimprinting and dip coating.

[0048] The vaccines employed in the present invention may contain live, attenuated, modified or killed microorganisms or their toxins or tumor antigens which when administered into the body stimulate the body’s immune system to produce antigen -specific antibodies.

[0049] The amount of antigen used in the devices and methods of the present invention include amounts necessary to provide an immune response. At least one dose selected from the group consisting of a Ipg dose, 2pg dose, 3pg dose, 4pg dose, 5pg dose, 6pg dose, 7pg dose, 8pg dose, 9pg dose, lOpg dose, 15pg dose, 20pg dose, 25pg dose and a 30pg dose may be sufficientto induce an immune response in humans. The dose of the antigen may be administered to the human within a range of doses including from about Ipg to about 50pg, from about Ipg to about 30pg, from about Ipg to about 25 pg, from about I g to about 20pg, from about I g to about 15 g, from about Ipg to about 10 pg, from about 2 pg to about 10 pg, from about 2 pg to about 8pg, from about 3 pg to about lOpg, from about 3 pg to about 8pg, from about 3 pg to about 5pg, from about 4pg to about lOpg, from about 4pg to about 8pg, from about 5pg to about lOpg, from about 5pg to about 9pg, and from about 5pg to about 8pg.

[0050] The amount of antigen used in the devices and methods of the present invention include amounts necessary to provide an immune response and may include 1000 to 10000 or 1000 to 9000 or from 1000 to 8000 or from 1000 to 7000 or from 1000 to 5000 or from 1000 to 4000 or from 1000 to 3000 or from 1000 to 2000 or from 2000 to 10000 or 2000 to 9000 or from 2000 to 8000 or from 2000 to 7000 or from 2000 to 5000 or from 2000 to 4000 or from 2000 to 3000 or CCID50 of both M and R each loaded onto the HD-MAP.

[0051] The present invention also relates to devices, formulations and methods for increasing the stability of vaccine formulations including but not limited to influenza and inactivated polio vaccine due to the use of excipients which include but are not limited to cyclodextrins, amino acids, reducing agents carbohydrates and proteins and combinations thereof. Excipients include but are not limited to Histidine, Sodium acetate, Sodium chloride, Sodium citrate, Sodium phosphate, Sodium sulfate, Sodium succinate, Gelatin, Hydrolysed Gelatin,, Protamine sulfate, Arginine, Aspartic acid (sodium salt), Glutamic acid, Glycine, Isoleucine, Lactic acid, Lysine, Maleic acid, Malic acid (sodium salt), Methionine, Urea, EDTA, Magnesium chloride, Benzalkonium chloride, Brij 35, Poloxamer 188 (Pluronic F-68), Polysorbate 20, Polysorbate 80, Sodium docusate, Triton X-100, Lactose, Sucrose, Trehalose, Glycerol, Mannitol, Sorbitol, Gamma-Cyclodextrin, 2-OH propyl b-CD, Sulfobutyl ether betacyclodextrin, Carboxymethyl cellulose, Dextran sulfate, Dextran 40, PEG-3350, Sodium Hyaluronate, Sodium thioglycolate, Cysteine, and Glutathione and combinations thereof.

[0052] The measles rubella compositions of the present invention may contain 0.5 to 5% or 0.5 to 4% or from 0.5 to 3% or 0.5 to 2% w / v sorbitol, lOmM to lOOmM or 10 mM to 75mM or 10 to 50 mM L-histidine, 0.5 to 4% or 0.5 to 3% or from 0.5 to 2% or 0.5 to 1% w / v trehalose dihydrate w / v, ImM to lOmM or ImM to 7.5mM or ImM to 6mM or from ImM to 4mMsodium phosphate dibasic dihydrate, ImM to lOmM or ImM to 7.5mM or ImM to 6mM or from ImM to 4mM sodium phosphate monobasic dihydrate and optionally 0.5 to 4% or 0.5 to 3% or from 0.5 to 2% or 0.5 to 1% w / v hydrolyzed porcine gelatin. The pH of the composition may be from pH 6.5 to 7.0 or from 6.6 to 6.9.

[0053] Stability of vaccine compositions and components can be measured by a loss in antigen potency. This loss in potency can be determined under a variety of conditions, such as storage temperature and storage humidity at various time points. Typically, vaccines which are in solution are stored at 4°C or at room temperature (about 25°C). It would be preferable to be able to store vaccine at at least room temperature or higher temperatures (35°C - 45°C) such that cold storage would be unnecessary.

[0054] The methods and compositions of the present invention provide microprojection arrays that can be coated with multiple incompatible vaccine antigens that are stable over time. The vaccine compositions of the present invention are stable at at least 2-8 °C for at least 1 or at least 2 or at least 3 or at least 4 or at least 5 or at least 6 or at least 7 or at least 8 or at least 9 or at least 10 or at least 12 or at least 13 or at least 14 or at least 15 or at least 16 or at least 17 or at least 18 or at least 19 or at least 20 or at least 21 or at least 22 or at least 23 or at least 24 or at least 30 or at least 36 months at various temperatures and conditions. The stability of the vaccine formulations may be measured by a variety of techniques including but not limited to ELISA and SDS-PAGE silver stain.

[0055] The methods and compositions of the present invention provide microprojection arrays that can be coated with multiple incompatible vaccine antigens that are stable over time. The vaccine compositions of the present invention are stable at at least 25 °C for at least 1 or at least 2 or at least 3 or at least 4 or at least 5 or at least 6 or at least 7 or at least 8 or at least 9 or at least 10 or at least 12 or at least 13 or at least 14 or at least 15 or at least 16 or at least 17 or at least 18 or at least 19 or at least 20 or at least 21 or at least 22 or at least 23 or at least 24 or at least 30 or at least 36 months at various temperatures and conditions. The stability of the vaccine formulations may be measured by a variety of techniques including but not limited to ELISA and SDS-PAGE silver stain.

[0056] To evaluate vaccine stability following drying, antigen values of recovered vaccine were determined using the ELISA assay. The percent potency of recovered dried vaccine was calculated by normalizing the antigen values of recovered dried samples to the values of an inliquid stock vaccine stored at 2-8°C, which was considered to have 100% potency. The drying potency loss was calculated by subtracting the percent potency of freshly dried vaccine samples (recovered immediately after drying) from the in-liquid stock vaccine stored at 2-8°C (i.e., 100% - relative percent potency after drying = drying potency loss). Similarly, the storage potency loss was determined by subtracting the relative potency of the stored samples with the relative percent potency of the sample recovered immediately after drying (i.e., 100% - relative percent potency after storage - relative percent potency after drying = storage potency loss).

[0057] Reduction of potency for the formulations / antigens of the present invention upon rapid drying can be about 0% or less than about 5% or less than about 10% or less than about 15% or less than about 20% or less than about 25% or less than about 30% or less than about 35% or less than about 40% or less than about 45% or less than about 50% or less than about 55% or less than about 60% or less than about 65% or less than about 70% or less than about 75% or less than about 80% or less than about 85% or less than about 90%.

[0058] Reduction of potency for the formulations / antigens of the present invention upon rapid drying and storage at at least 2-8°C for at least 1 or at least 2 or at least 3 or at least 4 or at least 5 or at least 6 or at least 7 or at least 8 or at least 9 or at least 10 or at least 12 or at least 13 or at least 14 or at least 15 or at least 16 or at least 17 or at least 18 or at least 19 or at least 20 or at least 21 or at least 22 or at least 23 or at least 24 or at least 30 or at least 36 months can be about 0% or less than about 5% or less than about 10% or less than about 15% or less than about 20% or less than about 25% or less than about 30% or less than about 35% or less than about 40% or less than about 45% or less than about 50% or less than about 55% or less than about 60% or less than about 65% or less than about 70% or less than about 75% or less than about 80% or less than about 85% or less than about 90%.

[0059] Reduction of potency for the formulations / antigens of the present invention upon rapid drying and storage at at least 25 °C for at least 1 or at least 2 or at least 3 or at least 4 or at least 5 or at least 6 or at least 7 or at least 8 or at least 9 or at least 10 or at least 12 or at least 13 or at least 14 or at least 15 or at least 16 or at least 17 or at least 18 or at least 19 or at least 20 orat least 21 or at least 22 or at least 23 or at least 24 or at least 30 or at least 36 months can be about 0% or less than about 5% or less than about 10% or less than about 15% or less than about 20% or less than about 25% or less than about 30% or less than about 35% or less than about 40% or less than about 45% or less than about 50% or less than about 55% or less than about 60% or less than about 65% or less than about 70% or less than about 75% or less than about 80% or less than about 85% or less than about 90%.

[0060] The MR vaccines of the present invention may have a seroconversion rate for neutralizing antibodies with respect to measles of between 10 to 50% or from 10 to 40% or from 10 to 30% or from 10 to 20% for subjects that were previous exposed to measles either by contracting the disease or by previous vaccination. The MR vaccines of the present invention may have a seroconversion rate for neutralizing antibodies with respect to rubella of between 10 to 50% or from 10 to 40% or from 10 to 30% or from 10 to 20% for subjects that were previous exposed to rubella either by contracting the disease or by previous vaccination.

[0061] In preferred embodiments the microprojections of the microprojection array are coated by an aseptic print-head type device which rapidly provides small droplets which dry quickly on the microprojections. In preferred embodiments the coating such as a vaccine formulation rapidly dries on the top portion of the microprojection to increase the amount of vaccine that can be delivered. The aseptic print head device may deliver multiple drops to the microprojections either sequentially or in an alternating fashion. In one embodiment of the print head device the device comprises the housing is connected to the pumping chamber where the fluid to be dispensed is stored. The fluid flows into the pumping chamber through one or more ports and fluid is pushed by the plate membrane out through the nozzles in the nozzle plate to be distributed onto the microprojections. The housing may have ports for conducting fluid into the pumping chamber. The unimorph PZT impacts the plate membrane which is held in place by a restrictor plate. All of these parts are assembled with the housing and the descender plate and nozzle plate. The embodiments utilizing the unimorph PZT are assembled using a bio-compatible epoxy.

[0062] The microprojections of the array of the present invention may be of any shape including cylindrical or conical. Other geometries are also possible. The microprojection arrays may have substrate with a plurality of microprojections protruding from the substrate whereinthe microprojections have a tapering hexagonal shape and comprise a tip and a base wherein the base has two substantially parallel sides with a slight draught angle of approximately 1 to 20 degrees up to a transition point at which point the angle increases to from about 20 degrees to about 70 degrees. A sharp blade-like tip will allow for enhanced penetration of the microprojections into the skin while also generating an enhanced localized cell death / bystander interaction in the skin with a different profile than conical microprojection arrays. In a preferred embodiment the microprojections are made of a polymer and are slightly blunted at the tip with shoulders near the tip on which the coating material may attach such that the coating material does not drip down the microprojection and onto the base of the microprojection array.

[0063] In the present invention the density of the microprojections is relatively high which means the microprojections are spaced relatively close together. The density of the microprojection on the microprojection arrays may be about 1000 microprojections / cm2, or about 1500 microprojections / cm2, or about 2000 microprojections / cm2, or about 2500 microprojections / cm2, or about 3000 microprojections / cm2, or about 3500 microprojections / cm2, or about 4000 microprojections / cm2. The density of the microprojection on the microprojection arrays may be from about 500 to about 5000 microprojections / cm2, or from about 500 to about 4000 microprojections / cm2, or from about 500 to about 3000 microprojections / cm2, or from about 500 to about 2000 microprojections / cm2, or from about 5000 to about 71500 microprojections / cm2, or from about 500 to about 1000 microprojections / cm2.ExamplesExample 1Stability of Measles and Rubella

[0064] An excipient screen was conducted with the excipients listed below for measles and rubella as determined by an extracted genome assay and a Triton treated method. In the Triton screen, 1% Triton was used to lyse the virus envelope and release genomic RNA prior to qt- PCR quantitation. In the RNA extracted method a guanidine lysis and silica membrane (QIAmp) based extraction method which disrupts measles and rubella virus nucleocapsid and purifies RNA larger than 200 base pairs was employed.

[0065] Excipients: 50mM histidine, 1% trehalose, 0.1M histidine, 1% hydrolysed gelatin, 1.6% sucrose, 0.05M arginine + 0.05M glutamic acid, 50mM sodium phosphate, 0.1M arginine, 0.045M arginine + 0.045M glutamic acid, 0.2% sodium hyaluronate, 0.3% carboxymethyl cellulose, 2% sorbitol, 2% lactose, 1% HAS, 0.05% polaxamer 188, 1% BSA, 2% mannitol, 1% PEG-3350, 0.2% protamine sulfate, 50mM sodium citrate, lOmM magnesium chloride, ImM EDTA, 150mM sodium sulfate, 50mM sodium succinate, 0.1M glutamic acid, 60M Tris, 0.1M lactic acid, 4.5% sulfobutyl ether beta-cyclodextrin, 150mM sodium chloride, 0.1M lysine, 1% dextran-40, ImM DTT, 5% gamma-cyclodextrin, 0.1M aspartic acid, 0.1M urea, 0.1M maleic acid, 0.04% benzalkonium chloride and 2% sorbitol + 50mM phosphate + 1% hydrolysed gelatin + 1% trehalose.

[0066] During the preparation of samples for the four week stability study described above an additional plate was included in the MR samples incubated at 4°C, which covered approximately 50% of the excipient combinations tested in the study (Table 1).Table 1 - Excipient combinations

[0067] The excipients and viruses were tested on dried discs which comprised 0.25pL of measles virus bulk, 0.25pL of rubella virus bulk and 9.5pL of excipients in lOmM phosphate buffer (PB) (pH 6.8) or PB alone. The excipient combinations are were tested at To and at 40°C for 3 and 7 days and at 37°C for 1 and 2 weeks and at 25°C at 2 weeks and are tested at 1, 2, 3, 4, 5 and 6 months at 25°C.

[0068] The results demonstrate that: 1) To liquid measles / rubella in the eight formulation showed comparable potency to -80°C bulk; 2) when dried and stored at 37°C up to 2 weeks or40°C for 1 week the MeV was more stable in the histindine + gelatin + trehalose formulations except when combined with arginine; and 3) RuV was stable in all formulationsExample 2Stability Study of Clinical Lots

[0069] Clarified virus pools for both M and R were supplied by Serum Institute of India and processed. The clarified pools were combined to produce a combined MR bulk which was concentrated by tangential flow filtration so that the required dose could be loaded onto HD- MAPs, and to exchange the harvest buffer to one containing excipients (sorbitol 2%w / v, L- histidine 50mM, trehalose dihydrate 1% w / v, sodium phosphate dibasic dihydrate 5.4mM, sodium phosphate monobasic dihydrate 4.6mM, and hydrolyzed porcine gelatin 1%) adjusted to pH 6.8 by 10% HC1 to stabilize M and R vaccines on HD-MAPs. MR-Vac (Batch 0090N001B, expiry June 2022) was used as SC control. The potency of MR-Vac was determined to be 1,300 CCID50 measles per dose and 5,200 CCID50 rubella per dose.Table 2 - Excipient formulation

[0070] In addition to this, 1% hydrolysed gelatin w / v (from SIIL) is added after the concentration step. The step also concentrates gelatin from the starting material, so the gelatin concentration is best described as no less than 1% hydrolysed gelatin (w / v).

[0071] HD-MAPs were manufactured by injection moulding of a polymer, to produce HD- MAPs of 10.6 x 10.6 mm with approximately 1,600 projections per MAP. Each projection was approximately 350 pm high and 120 pm wide at the base. Vaccine was aseptically applied tothe tips of each projection of gamma-irradiated (> 25kGy, Steritech, Australia) HD-MAPs using the ‘M-jet’ process developed by Vaxxas. HD-MAPs were produced to deliver a single dose-level of > 1,000 CCID50 of each virus (M and R) per HD-MAP. The doses cited throughout this report refer to the estimated delivered dose. Preparatory studies using ex vivo and in vivo pig-skin assays determined the delivery efficiency of this MAP-vaccine combination to be approximately 50%; therefore, to deliver the desired dose, > 2,000 CCID50 of both M and R were loaded onto each HD-MAP. After HD-MAP manufacture, the actual delivered doses were estimated to be: 3,100 and 4,300 CCID50 per MAP for M and R respectively. After coating with MR vaccine, the HD-MAPs were contained within an integrated applicator containing a dome-spring with a foil seal covering the skin-facing side of the applicator and packed in a foil pouch.

[0072] MR-coated HD-MAPs from the clinical batch were stored at 2-8 °C for various timepoints up to 24 months (study ongoing). At the initial (TO), 12 month, and 24 month timepoints, five HD-MAPs that had been stored at 2-8 °C were transferred to 40 °C for three days to mimic conditions required for controlled temperature chain (CTC) qualification. Relative humidity for the 40 °C condition was 60 % for TO and 12 month and 75 % RH for 24 month testing. At each timepoint, the coating was eluted from the HD-MAPs (n=5) in 300 pl elution buffer (serum-free tissue culture media), then tested in the CCID50 potency assay. Prior to manufacture for the clinical trial, another thermostability study was conducted at 2-8 °C for various timepoints up to 30 months (study ongoing) and 25+5 °C for 12 months. Accelerated testing of 3 days at 40 °C (60 % RH), 7 days at 37°C (60 % RH) and 14 days at 37°C (60 % RH) were also included. Lyophilized MR- Vac was also included in accelerated and long-term conditions in this pre-clinical study. CCID50 assay was performed using Vero (ATCC CCL- 81) cells incubated for 6 days at 37 °C and RK13 (ATCC CCL-37) cells incubated for 10 days at 31 °C post-titration and inoculation of samples, for the detection of measles and rubella respectively. Cytopathic effect (CPE) was visually assessed after incubation and titer calculated using the Spearman-Karber method. Biological reference pathogen for each virus was used to assess assay validity for each set of testing. Simple linear regression was performed to trend data, plotted with a 95% confidence band (GraphPad Prism 9.5.0).

[0073] Real-time stability studies showed minimal loss of potency of M or R viruses, after storing MR HD-MAPs (clinical batch) at 2-8 °C for 24 months, with a degradation rate of 0.004 and 0.008 logCCIDso per MAP per month for measles and rubella, respectively (Figure 1A, 1C). Under CTC conditions of three days at 40 °C, 60 %RH, MR HD-MAPs (clinical batch) showed minimal loss (up to 0.28 logCCIDso / virus / MAP) compared to MAPs stored at 2-8 °C assayed in parallel and still met minimum potency specifications (3.3 logCCIDso / virus / MAP) under all temperature conditions at TO, 12, and 24 months (Figure IB, ID). Susceptibility to potency loss at 40 °C was similar despite length of storage at 2-8 °C.

[0074] At nine months, rubella testing did not meet assay validity criteria and insufficient MAPs were available to repeat testing. At TO and 12-months, five MAPs were stored at 40 °C, 60 % RH for three days prior to testing (grey bar), then assayed for measles and rubella potency in parallel with MAPs stored at 2-8 °C. The log loss relative to 2-8 °C MAPs assayed in parallel is shown above the bar. For all graphs, minimum specification (3.3 logCCIDso per virus per MAP) is shown as a dotted line. Linear regression was performed and 95% confidence bands are shown.

[0075] In pre-clinical stability studies the rubella virus was more stable under all conditions in both MR-Vac and HD-MAPs, in comparison to measles virus. Overall, MR HD-MAP stability for measles potency was improved on HD-MAPs, particularly for 14-day storage at 37 °C (60 %RH). Overall, stability of rubella was comparable between MR-Vac and HD-MAP. LogCCIDso loss was lower under some conditions for HD-MAPs compared to MR-Vac (7 days, 37 °C) but higher under others (30 months, 2-8 °C). Only small losses were detectable for both MR HD-MAPs and MR-Vac (up to 0.24 logCCIDso). The clinical MR HD-MAP stability program focused on long-term 2-8 °C storage and did not include 25 °C or accelerated testing beyond 7 days, as performed for the pre-clinical study, nor comparison to MR-Vac.Table 2 - LogCCIDso loss for MR HD-MAPs and MR-Vac (pre-clinical studies prior to clinical batches). Values are logCCIDso loss, compared to 2-8 °C samples assayed in parallel (for accelerated conditions) or compared to TO (for long-term conditions at 2-8 or 25 °C). All accelerated conditions were performed at 60 % RH.

[0076] At 24 months for the pre-clinical MR HD-MAPs and 6 months for the clinical MR HD- MAPs, other product attributes were also assessed including sterility, applicator performance, and vaccine coating appearance. All testing met specifications set for product release into trials.Example 3Clinical Trial

[0077] A randomized, partially double-blind, placebo -controlled trial was conducted in which clinical staff and participants were blind as to which HD-MAP treatment was administered. All laboratory investigators were blind to treatment and participant allocation. The primary objective was to measure the safety and tolerability of MR vaccines delivered by HD- MAP in comparison to an uncoated HD-MAP and SC injection of a MR vaccine (MR-Vac, Serum Institute of India Ltd, Pune, India). Exploratory outcomes were to evaluate the immune responses to HD-MAP application by foci-reducing neutralization titre (FRNT) and IgG ELISA.

[0078] Healthy males and females (non-pregnant and non-nursing) aged 18-50 years, with a BMI in the range of 18-32 kg / m2(N = 63), were recruited and randomly allocated into one offour vaccination groups with > 15 participants per group. Randomization was pre-determined, and sealed participant-specific code break envelopes were produced by the statistician responsible for preparing the randomization. The randomization was provided to the unblinded pharmacist for re-labelling the investigational products. The four treatment groups were: placebo (uncoated HD-MAPs); low dose, (-3,100 and -4,300 CCID50 measles and rubella, respectively); high dose, (-9,300 and -12,900 CCID50 measles and rubella, respectively. SC control (> 1,000 CCID50 per virus). MR Vac tested during the study was determined to contain 1,300 and 5,200 CCID50 measles and rubella per dose. The sample size was not based on any formal statistical calculations, as is typically the case for Phase I vaccination studies. However, the 15 participants in a group would have an 80% probability of showing at least one adverse event if the true rate of that event was more than 10.2%, and over the 45 participants receiving any MAP there was an 80% probability of showing at least one adverse event if the true rate of that event was more than 3.6%.

[0079] Three HD-MAPs were applied to all HD-MAP recipients. The high and low doses were achieved by applying three or one MR-coated HD-MAPs to a participant, plus none or two uncoated HD-MAPs. Participants in the placebo group received three uncoated HD-MAPs.

[0080] Participants were vaccinated on day 0. Application sites were selected to be free from scarring, tattoos, skin conditions, sunburn, and heavy hair. The area for application was marked and photographed. The foil seal on the HD-MAP was removed and the device was applied to the skin of the upper arm overlying the deltoid muscle. A slight pressure was applied to the top of the HD-MAP applicator device to activate the internal dome spring that propels the HD- MAP to the skin. The device was held in place for 60 seconds before being removed. All applications were performed by trained study team members.

[0081] Participants were monitored by clinic safety assessment visits at days 3, 7, 28, and 56; and phone calls at days 1 and 14. On day 0, all vaccination sites were assessed at prevaccination, 10 minutes, 1, and 2 hours after HD-MAP or SC administration. Photographs of the treatment sites were taken at every clinic review. Skin reactions were assessed for erythema, oedema, induration, tenderness, bruising, skin flaking, visibility, itching, and bleeding.

[0082] Serum blood samples were collected from participants at day 0 (pre-dose), 7, 28 and 56 post-vaccination. Aliquots of serum were prepared using serum separation tubes and stored at -80 °C until analysis.

[0083] For analysis of measles and rubella IgG titres serum was measured at Sullivan Nicolaides Pathology. For measles IgG, a chemiluminescence immunoassay was run using the Liaison KL instrument (measles IgG kit), and for rubella IgG, a two-step chemiluminescent microparticle immunoassay was run using the Abbott Architect i2000 instrument. The result for measles IgG (AU / mL) was Negative if <13.50, Equivocal if from 13.50 to 16.49 and Positive if >16.50. The result for rubella IgG (lU / mL) was Negative if <5.0, Grayzone if 5.0 to 9.9, Low Positive if 10 to 20 and Positive if >20.

[0084] A foci-reduction neutralization (FRN) assay was performed by 360biolabs for each virus to measure functional antibodies against measles and rubella. Briefly, heat-inactivated human serum was titrated and mixed with a set concentration of measles or rubella virus (vaccine strains, equivalent to 50 foci forming units inoculated per well) and incubated for 2 hours at 37 °C. Following this, serum / virus solutions were inoculated onto Vero monolayers prepared one day prior. After a one-hour incubation, overlay media of 0.5 % carboxymethylcellulose in 2% FBS MEM was added to all wells. Assay plates were incubated for 2 days at 37 °C or 5 days at 33 °C for measles and rubella, respectively. After the incubation period, plates were fixed with ice-cold acetone then immunostained using anti-measles nucleoprotein ms antibody (Abeam 106292) and anti-ms IgG HRP (Abeam 97023) for antimeasles and anti-rubella capsid antibody.

[0085] For neutralising antibodies, the two main analyses were two linear mixed regression models, one for measles and one for rubella, assessing for a change in log titre values from baseline. Categorical predictors were treatment group, visit (Days 7, 28 and 56), and a group by visit interaction term. Baseline log titre value was also included as a continuous predictor. Titre values were modelled on the log-scale so that the model residuals were normally distributed. The reference categories to which results were compared were the Active control group (vs other treatment groups) and Day 7 (vs Day 28 and 56). Models were fit using restricted maximum likelihood estimation, with the exception of the likelihood ratio test models, which were generated using maximum likelihood estimation. A compound symmetrycovariance structure was used. Analyses were performed in SAS version 9.4, plots were prepared in GraphPad Prism version 9.5.0.

[0086] Nearly all of the participants had protective levels of measles and rubella neutralizing antibodies before vaccination (Figures 2A and 2B and Table 3).Table 3 - Titer and seroconversion comparison of the study conditions over time

[0087] There was no significant increase in FRNT or ELISA titres and no seroconversions against measles or rubella in the uncoated MAP group after vaccination. In contrast, FRNT titres against measles and rubella significantly increased in all active groups following vaccination, peaking at day 28 post vaccination. Titres declined slightly by day 56 but remained above baseline. At Day 28, titres for all active groups were significantly above placebo (p < 0.05) for both viruses, nor was there a statistically significant difference between MAP groups and MR Vac (p < 0.05) for both viruses. Seroconversion rates for neutralizing antibodies against measles at day 28 were: 37.5%, 18.8% and 35.7% in the low-dose HD-MAP, high-dose HD-MAP and MR- Vac groups respectively. The corresponding seroconversion rates for rubella neutralizing antibodies were 37.5%, 25.0% and 35.7%.

[0088] The fold-increase in titre, and therefore the seroconversion rates, were dependent on the pre-vaccination titre, with four-fold increases only being seen in participants with prevaccination titres < 580 mIU / ml for measles (Figure 3). For measles, there were 9, 4 and 8 such subjects in the low-dose, high-dose and MR- Vac groups respectively, including the 6, 3 and 5 subjects that seroconverted in each group respectively (Figure 3A). A similar pattern was seen for rubella. Across all groups, 15 subjects seroconverted for rubella antibodies at day 28; of these, 12 had pre-vaccination titres of < 32 lU / ml. At DO, 9 subjects in each group had a titre < 32 lU / mL for Rubella (Figure 3B).

[0089] Anti-measles and anti-rubella IgG measured by ELISA showed a similar pattern of response to neutralizing antibodies. Antibody concentrations peaked at day 28 and decreased slightly by day 56. As with neutralizing antibodies, the fold-increase in titer was dependent on the baseline, pre-vaccination titer with greater fold increases being seen in participants with low IgG concentrations pre-vaccination.

[0090] There were no life threatening or serious treatment emergent adverse events (TEAEs), no TEAEs resulting in study withdrawal and no TEAEs resulting in death. TEAEs (28) deemed related to study treatment were experienced by 20 (31.7%) subjects, with 16 (25.4%) subjects experiencing 19 localized study treatment-related TEAEs and 7 (11.1%) subjects experiencing 9 systemic study treatment-related TEAEs. Most adverse events were mild or moderate in severity, with only 1 (1.6%) subject in the active control group (MR-Vac) experiencing a severe TEAE (gastroenteritis).

[0091] Some HD-MAP application sites remained visible at Day 7, with visibility noted in 48 (100.0%) sites for uncoated HD-MAP, 46 (95.8%) sites for low dose HD-MAP and 48 (100.0%) sites for high dose HD-MAP. By Day 28 and Day 56, a minority of application sites were visible for uncoated and low dose HD-MAP, while 46 (95%) and 26 (54.2%) of sites were visible in high-dose HD-MAP recipients at days 28 and 56 respectively. In contrast, for MR-Vac, 13 (86.7%) sites were visible at 10 minutes post-injection but by day 3, only 3 (20.0%) sites were visible. By day 7, none of the 15 injection sites were visible for the MR- Vac.

[0092] This trial was the first clinical trial of a live attenuated virus vaccine administered using the HD-MAP. The MR HD-MAP administration was well tolerated and induced immune responses similar to those seen with SC injection. In addition, MR vaccines coated onto HD- MAPs were at least as thermostable as standard, lyophilized vaccine.

[0093] The seroconversion rates to measles and rubella in this trial were relatively low regardless of delivery method. This is most likely due to the participants having high antibody titres at baseline. Other trials of novel delivery devices for measles vaccines have also found that seroconversion rates were inversely correlated with baseline titre, and seroconversion rates were in the range of 7 - 17% for measles when participants with high starting titres were included (MVDP author group, Cape S, Chaudhari A, Vaidya V, Mulay R, Agarkhedkar S, et al. Safety and immunogenicity of dry powder measles vaccine administered by inhalation: a randomized controlled Phase I clinical trial. Vaccine 2014;32:6791-7. https: / / doi.Org / 10.1016 / j.vaccine.2014.09.071). A better indication of the immunogenicity of MR HD-MAPs will be provided from trials in naive subjects.

[0094] A previous study of transcutaneous (TC) delivery using skin abrasion followed by application of a projection-free MAP found that the TC delivery induced cell-mediated and mucosal immunity, but was a poor inducer of neutralising or ELISA antibodies in the serum. Cell-mediated and mucosal immunity were not measured in this trial, so it is not known whether HD-MAPs are strong inducers of these arms of the immune response, in addition to the serum antibody responses that were detected.

[0095] The local reactogenicity seen with MR HD-MAPs was similar to that seen in a Phase I trial with influenza vaccine and might be at least in part due to the fact that recipients had been vaccinated previously with MR vaccines. All reactions resolved completely, but this took weeks rather than days. The ability of MAPs to decrease the pain and anxiety associated with immunization was a major expectation of caretakers participating in a usability study.

[0096] MR vaccines have been shown to have improved thermostability on other MAP formats compared with the standard, lyophilized presentation. The controlled temperature excursion data indicates that MR HD-MAPs is suitable for use in the controlled temperature chain (CTC), facilitating their use in outreach settings.

[0097] This trial was the first clinical trial using the HD-MAP as integrated device, combining the MAP and the single-use applicator. It was also the first HD-MAP trial with a live-attenuated vaccine.

[0098] Within this disclosure, any indication that a feature is optional is intended provide adequate support (e.g., under 35 U.S.C. 112 or Art. 83 and 84 of EPC) for claims that include closed or exclusive or negative language with reference to the optional feature. Exclusive language specifically excludes the particular recited feature from including any additional subject matter. For example, if it is indicated that A can be drug X, such language is intended to provide support for a claim that explicitly specifies that A consists of X alone, or that A does not include any other drugs besides X. "Negative" language explicitly excludes the optional feature itself from the scope of the claims. For example, if it is indicated that element A can include X, such language is intended to provide support for a claim that explicitly specifies that A does not include X. Non-limiting examples of exclusive or negative terms include "only," "solely," "consisting of," "consisting essentially of," "alone," "without", "in the absence of (e.g., other items of the same type, structure and / or function)" "excluding," "not including", "not", "cannot," or any combination and / or variation of such language.

[0099] Similarly, referents such as "a," "an," "said," or "the," are intended to support both single and / or plural occurrences unless the context indicates otherwise. For example "a dog" is intended to include support for one dog, no more than one dog, at least one dog, a plurality of dogs, etc. Non-limiting examples of qualifying terms that indicate singularity include "asingle", "one," "alone", "only one," "not more than one", etc. Non-limiting examples of qualifying terms that indicate (potential or actual) plurality include "at least one," "one or more," "more than one," "two or more," "a multiplicity," "a plurality," "any combination of," "any permutation of," "any one or more of," etc. Claims or descriptions that include "or" between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context.

[0100] Where ranges are given herein, the endpoints are included. Furthermore, it is to be understood that unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or subrange within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0101] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention.

[0102] While this invention has been particularly shown and described with references to example embodiments thereof, it will be understood by those skilled in the art that the various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.

[0103] Throughout this specification and claims which follow, unless the context requires otherwise, the word “comprise”, and variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated integer or group of integers or steps but not the exclusion of any other integer or group of integers. As used herein and unless otherwise stated, the term "approximately" means ±20%.

Claims

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:1) A vaccine composition for coating a microprojection array having two or more microprojections comprising measles virus and rubella virus and sorbitol, L-histidine, trehalose dihydrate sodium phosphate dibasic dihydrate, sodium phosphate monobasic dihydrate and hydrolyzed porcine gelatin.2) The vaccine composition of claim 1 wherein the composition contains 0.5 to 5% w / v sorbitol, lOmM to lOOmM L-histidine, 0.5 to 4% w / v trehalose dihydrate w / v, ImM to lOmM sodium phosphate dibasic dihydrate, ImM to lOmM sodium phosphate monobasic dihydrate and 0.5 to 4% w / v hydrolyzed porcine gelatin.3) The vaccine composition of claim 1 wherein the composition contains 2% w / v sorbitol, 50mM L-histidine, 1% w / v trehalose dihydrate w / v, 5.4mM sodium phosphate dibasic dihydrate, 4.6mM sodium phosphate monobasic dihydrate and 1% w / v hydrolyzed porcine gelatin.4) The vaccine composition of claim 1 wherein the composition has a pH of between 6.5 and 7.0.5) The vaccine composition of claim 1 wherein the amount of rubella and measles virus is at least 1000 CCID50 for each virus.6) The vaccine composition of claim 5 wherein the amount of rubella virus is between 3000 to 5000 CCID50 and the amount of measles virus is 2000 to 4000 CCID50.7) The composition of claim 1 wherein the composition is sprayed onto the microprojections of the microprojection array and the composition dries on the microprojections.8) The composition of claim 1 wherein the microprojection array comprises a base and solid microprojections which project from the base and wherein the microprojections are 300 to 400pm in length.9) The composition of claim 1 wherein the microprojection array is made from a polymer.10) The composition of claim 9 wherein the polymer is a liquid crystal polymer.11) The composition of claim 8 wherein the projections extending from the base wherein the projections comprise a targeting section for delivering the viruses to thereby cause an immunological response and a cylindrical support section for supporting the targeting section, wherein two or more steps exist between the targeting section and the support section and the support section is wider than the targeting section.12) The composition of claim 7 wherein the composition is stable for at least 3 months.13) The composition of claim 7 wherein the composition is stable for at least 6 months.14) The composition of claim 7 wherein the composition is stable for at least 12 months.15) The composition of claim 7 wherein the composition is stable for at least 24 months.16) The composition of claim 7 wherein the composition is stable for at least 36 months.17) The composition of claim 12 wherein stability is measured by titer.18) The composition of claim 12 wherein stability is measured by genomic material.19) A method of immunizing a human against measles and rubella by administering to the human the composition of claim 7 by projecting the microprojection array into the human’s skin thereby penetrating the skin with the microprojections coated with the composition.20) The method of claim 19 wherein a seroconversion rate for neutralizing antibodies with respect to measles is between 10 to 50% for subjects that were previous exposed to measles either by contracting the disease or by previous vaccination.21) The method of claim 19 wherein a seroconversion rate for neutralizing antibodies with respect to rubella is between 10 to 50% for subjects that were previous exposed to rubella either by contracting the disease or by previous vaccination.

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