WO2025101515A1 - Compositions and methods for single injections - Google Patents

Abstract

Disclosed are injectable needles. Also disclosed are methods of delivering a therapy to a subject in need thereof using the injectable needles.

Description

COMPOSITIONSAND METHODS FOR SINGLE INJECTIONS

RELATED APPLICATION

This application claims the benefit of priority to U.S. Provisional Application No. 63/547,469, filed November 6, 2023; the contents of which are fully incorporated by reference herein in their entirety.

BACKGROUND

Numerous drugs and vaccines require multiple doses to provide sufficient treatment and protection. Similarly, lipid nanoparticle (LNP)-based mRNA vaccines for SARS-CoV-2 necessitate several doses at predetermined time intervals for effective immunizations. However, requiring multiple doses poses challenges of missed or mistimed doses and can lead to serious diseases and, in some cases, death. Considering that nearly half of the unimmunized children received at least one dose of vaccine, a delivery system that can mimic the current multiple bolus administrations in a single injection could significantly improve global immunization coverage. Thus, there is an ongoing unmet need for new compositions and methods for administering vaccines and other bioactive agents.

SUMMARY OF THE INVENTION

In certain aspects, disclosed herein are injectable needles for administering vaccine and other therapeutic agents that require multiple doses (e.g, vaccine boosters).

In one aspect, the present disclosure provides injectable needles comprising a needle body, a needle tip, a compartment, a first composition, and a second composition; wherein: the needle body and the needle tip are connected to one another; the first composition and the compartment are encapsulated by the needle tip and the needle body; the compartment comprises a compartment base and a compartment cap; and the second composition is encapsulated in the compartment.

The injectable needle may comprise a plurality of compartments (e.g., 2, 3, 4, or more compartments), wherein each compartment encapsulates a composition, and each compartment releases its encapsulated composition at a different point in time. Each of said compositions may be the same or different.

In another aspect, the present disclosure provides methods of delivering a therapy to a subject in need thereof comprising contacting the subject with an injectable needle disclosed herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGs. 1A & IB show schematic representations of a self-boosting lipid nanoparticle-based mRNA (mRNA-LNP) vaccination system.

FIG. 2 shows an exemplary design of an injectable needle disclosed herein.

FIGs. 3A & 3B show exemplary dissolvable needle tips and their mechanical properties. Different polymeric materials were screened to find the best mechanical properties. Once the polymer needles were made, we performed Instron compression test on the polymer needles to find the maximum force they can endure before breaking. The load cells were compressed at a 5mm/min rate and stopped at 40% load change and recorded the maximum load [N] before breakage. For all needle sizes, there was no significant difference in max load between materials 1 thru 4, but material 5 experienced breakage with less force. For needle gauges, 21 G and 23 G endured compression force a lot better than the other smaller needle sizes. However, since it takes only about 2 N to puncture human skin, and because all 5 materials endured force larger than 2N for 21 Gand 23G, we focused on designing the needle to be in that size range.

FIGs. 3C & 3D show further exemplary needle tips and their properties. Out of the five materials, Material #5 dissolved the fastest.

FIGs. 3E & 3F show the results of mRNA encapsulation in certain exemplary needle tips. We fabricated needles with five different vaccine inks (vaccine ink formulation) and redissolved them for running RiboGreen assay to assess LNP encapsulation efficiency (EE) and for running Femto Pulse RNA fragment analysis to assess mRNA integrity. With RiboGreen assay, we studied how well the LNP structure remain intact (=EE). Vaccine ink made with material 5 showed the highest EE in this study. With Femto Pulse RNA fragment analysis, we studied how well the mRNA fragments inside LNPs remain intact (=mRNA integrity). Vaccine ink made with material 5 showed the highest mRNA integrity in this study. FIG. 4 shows a schematic representation illustrating the importance of retractable injectable needles.

FIGs. 5A-5C show the dissolution of exemplary needles. We made a dissolvable needle with material 5 for the needle tip and material 3 for the needle body, and submerged the entire needle in water, the tip material dissolved and was gone after 5 minutes, but the base remained and held the intact MP after 15 minutes. We repeated this test but with ex vivo human skin. We administered the 2-step (material 5 and 3) fabricated MP-embedded dissolvable needle to ex vivo human skin and retracted the needle 5, 15, and 30 minutes after administration. Prior to insertion, the tip and body were observable. After 5 minutes, the needle tip was gone. After 15 minutes, the needle body still held the MP and is retractable. After 30 minutes, the MP is administered to the subcutaneous region of the human skin.

FIGs. 6A & 6B show the investigation of exemplary SEAL base materials. We screened different PLGAs (Poly(lactic-co-gly colic acid)) with different molecular weights (PLGA material names - 502H, 502, 503H, 505, 756S, 858S, AP22) for their mechanical integrity and moldability. For mechanical robustness study, we performed Instron tensile test with PLGA films to find the maximum force they can endure before breaking. Each film sample was about 2cm x 1cm and roughly lOOum thick. The samples were pulled at a 5mm/min rate and the maximum load [N] before breakage was recorded. PLGA films with lower molecular weight generally teared more easily, meaning they are less suitable as a protective shell material of the SEAL particle. There was no significant difference for higher molecular weight PLGA examples. We also performed a moldability test to qualitatively find which PLGA material has a better shape after particle molding. PLGAs with lower molecular weight were too brittle to form the elongated box shape. PLGA 858s exhibited the best moldability results.

FIGs. 7A & 7B show the investigation of exemplary SEAL base materials. Before selecting the right MP cap material, we first had to optimize the sealing technique for different PLGA caps because we need to seal MPs with different PLGA materials to test their release kinetics. The principle of sealing technique involves temporarily melting PLGA cap and attaching it to the PLGA base so that the two parts meet and fuse, thereby encapsulating the cargo inside. Each PLGA cap needs different sealing temperatures as each material has different melting points. The temperature and sealing technique were optimized for our cap material candidates. We also assess mRNA-LNP vaccine block’s stability after exposing them to sealing temperatures. It is important that the mRNA-LNP remain potent and active even after being exposed to heat. When exposed to 40, 50, and 60 °C, which is the range of temperature required for sealing, both EE and mRNA integrity remained good (i.e., no reduction compared to control group).

FIGs. 7C & 7D show the further investigation of exemplary SEAL base materials. We fabricated blocks with 100 nm fluorescent nanobeads to mimic the 100 nm mRNA-LNPs. We sealed them in MPs with 858s base + 6 different PLGA caps (502, 503H, 505, AP22, 756s, 858s) to study their release kinetics. To study release kinetics, we are using 2 techniques. Firstly, we incubate sealed MPs in PBS at 37C and continuously take images to visually see the release of the cargo. Second, we incubated sealed MPs in PBS at 37C and take supernatant and measure fluorescence to quantify the fluorescence beads released from the core.

FIG. 8 shows an exemplary design of an injectable needle disclosed herein.

FIG. 9A shows the workflow for the fabrication of an exemplary needle, such as that discussed in Example 4.

FIG. 9B shows an exemplary injectable needle. 2-doses are loaded in a single injectable needle. The first dose at the needle tip and the second in the CS MP, which is engineered to release the 2nd dose 4 weeks after injection.

FIGs. 10A & 10B show leakage tests of an exemplary needle. We incubated the needle in PBS at 37 °C to see if there was any immediate notice of leakage or crack, which would be indicated by early release of the dye in the core. When we measured the absorbance of the supernatant and nothing was observed for at least 20 days.

FIG. 11 shows the imaging of an exemplary needle once it had been injected into skin. With uCT, it was clearly visible with high resolution, but the downside of uCT is that patients would have to go in a micro-CT scanner. With ultrasound imaging, it was visible, but with a lower resolution. The positive side of ultrasound imaging is, though, they come as a hand-held device, so it’s more convenient for patients. These results are favorable toward the potential tracing and retracting feature of the product.

FIG. 12A shows the evaluation of exemplary vaccine compositions. We used EER% and mRNA integrity (i.e. intact mRNA%) for evaluation. The values on the X-axis refer to the ratio of PVP:PVA. PVP:PVA 0: 1: EER% = 84.8% (+/- 4.9%) & intact mRNA% = 97.7% (+/- 3.3%); PVP:PVA 1:2: EER% = 86.2% (+/- 5.3%) & intact mRNA% = 98.4% (+/- 2.1%); and PVP:PVA 1: 1: EER% = 78.8% (+/- 1.5%) & intact mRNA% = 97.2% (+/- 5.5%). FIG. 12B shows the evaluation of exemplary vaccine compositions. The values on the X- axis refer to the ratio of PVP:PVA. PVP:PVA 1:2 group presented higher total mRNA and deliverable mRNA.

FIG. 12C shows the evaluation of exemplary vaccine compositions. The values on the X- axis refer to the ratio of PVP:PVA. The polymer composition was PVA only (0:1). We used EER% and mRNA integrity (i.e. intact mRNA%) for evaluation. We found that EE recovery was improved as the ink ratio increased. mRNA:polymer 1:50: EER% = 66.3% (+/- 11.0%); mRNA:polymer 1: 150: EER% = 71.8% (+/- 10.9%); and mRNA:polymer 1 :320: EER% = 84.8% (+/- 4.9%) & Intact mRNA% = 97.7% (+/- 3.3%).

FIG. 12D shows the evaluation of exemplary vaccine compositions. The values on the X- axis refer to the ratio of PVP:PVA. Here, we focused on two ink ratio groups: 1:50 and 1 :320. Note that in this study, we used polymer ratio of PVP:PVA 1 :2. Consistent with previous slide, we observed a decrease in EE recovery for 1:50 group compared to 1:320 group. However, given that the volume of each block was fixed, using less polymer (in the case of 1 :50) allowed a higher density of mRNA-LNP complex, which therefore increased the total amount of mRNA included in each block. Further, we identified that 1:50 group presented higher deliverable mRNA mass.

FIG. 12E shows the evaluation of exemplary vaccine compositions. We characterized the two groups with and without PLGA contacting. The formulation used was ink ratio 1:320, and polymer ratio 0: 1 (PVA only). The results showed no significant difference regarding EE recovery and total mRNA mass for the two groups.

FIG. 12F shows the evaluation of exemplary vaccine compositions. Use of PLGA as contacting surface for blocks did not introduce significant change in EER for the mRNA-LNP complex and PVP:PVA 1:2 group showed higher deliverable mRNA mass. PVP:PVA 0:1 (PVA only): EER% = 80.6% (+/- 5.2%), mRNA mass = 1.34 (+/- 0.17) pg, and deliverable mRNA mass = 1.08 (+/- 0.13) pg; andPVP:PVA 1:2: EER% = 76.6% (+/- 4.9%), mRNA mass = 1.86 (+/- 0.30) ug, and deliverable mRNA mass = 1.43 (+/- 0.23) pg.

FIG. 12G shows the evaluation of exemplary vaccine compositions. In this study, we tested the effect of sealing step of the PLGA particles on mRNA-LNP complex. Briefly, the cap of a SEAL particle was heated to the glass-transition temperature of the corresponding PLGA polymer, and then the cap was brought to touch the base of the SEAL particle. The cap and base components of the SEAL particles were combined via heat transfer. The heated cap touched with the base for approximately 5 minutes. This step could potentially put heat stress on the mRNA- LNP complex in the vaccine block. Here we used PLGA 858s for bases, and PLGA 502 for caps. EE recovery, total mRNA mass, and encapsulated mRNA mass were used to evaluate the effect of sealing step. The results showed no statistically significant change in the three evaluated parameters for control blocks (without sealing step) and the blocks experienced the sealing step. Without sealing: EER% = 81.2% (+/- 3.7%); mRNA mass = 2.24 (+/- 0.39) pg, and deliverable mRNA mass = 1.81 (+/- 0.32) pg. With sealing: EER% = 82.3% (+/- 2.2%), mRNA mass = 2.01 (+/- 0.37) pg, and deliverable mRNA mass = 1.66 (+/- 0.30) pg.

FIG. 12H shows the evaluation of exemplary vaccine compositions. We also tested the effect of redissolving solution on mRNA-LNP complex. Briefly, to evaluate the mRNA-LNP stability, we needed to redissolve the dried vaccine blocks. Here, we studied the effect of solution used for redissolving. We selected PBS buffer and water, and evaluated the EE recovery. Surprisingly, we noticed that the LNP showed a significantly lower EE when redissolved in PBS. The EE remained considerably high when redissolved in water. Additionally, we studied the effect of redissolving solution on mRNA-LNP complex without polymer. Here, we used pipette to fill concentrated LNP solution (no polymer) to SEAL base and left it dry thoroughly. Next, we redissolved the dried LNP in PBS and water. Consistent with the case of core blocks, here we observed a drastic decrease in EE recovery when water was used for redissolving compared to water.

FIG. 121 shows the evaluation of exemplary vaccine compositions. We further explored the effect of pH of the redissolving solution on EE recovery.

Here, we noticed that low pH aqueous solution (pH = 5 in this experiment) preserved high EE recovery. When the pH reached and surpassed neutral condition (pH = 7 and 10), low EE recovery was observed.

FIG. 13 shows the results of treating a mouse with an exemplary injectable needle system.

FIGs. 14A & 14B show the results of in vivo expressions with different vaccine ink formulations.

FIGs. 15A & 15B show the characterization exemplary lipid nanoparticles.

FIGs. 16A-16C show the internal nanostructure of exemplary lipid nanoparticles. LNPs, composed of L5ZDOPE/Cholesterol/PEG, were made with different NP ratios. FIG. 16A shows the mean size as measured by DLS. LNP size decreased with increasing NP ratio. FIG. 16B shows the mRNA copy number in each NP ratio as calculated based on the molecular volume model discussed in the method section. The number of mRNA copies is inversely proportional to NP ratio. FIG. 16C shows the mRNA encapsulation efficiency. Encapsulation efficiency increased at a higher NP ratio.

FIGs. 17A-17D show the Fabrication and retrieval of mRNA-LNP loaded MAPs. FIGs. 17A & 17B show a Schematic of mRNA-loaded microneedle (MN) fabrication process. FIG. 17A Modular vaccine ink is prepared by mixing mRNA-LNPs with a mixture of PVP:PVA. FIG. 17B Vacuum drying steps involve dispensing the vaccine ink in MN PDMS mold and applying a vacuum through PDMS to load the polymer-vaccine solution into the microneedle mold. FIG. 17C show HR-SEM micrographs of transverse cross section of a MN tip fabricated of different mRNA/ polymer ratios and kept under vacuum during sample transfer and preparation. Increasing mRNA/polymer ratio reduces LNPs aggregation and improves their dispersion within polymer matrix. FIG. 17D shows Cryo-TEM micrographs of mRNA-LNP in ink at different mRNA/ polymer ratios showing the morphology of mRNA-LNP before drying and after redissolution. FIG. 17E shows the LNP diameter after re-dissolution derived from analysis of cryo-TEM images.

FIGs. 18A-18F show mRNA Integrity. FIG. 18A shows LNP diameter (number average DLS diameter) decreased with increasing polymer: mRNA ratio. FIGs. 18A-18E show total mRNA recovery (FIGs. 18B-18D) and encapsulation efficiency (FIG. 18E) of LNPs of different NP ratios 1.5 (FIG. 18B), 3.1 (FIG. 18C) and 5.4 (FIG. 18D) embedded in different polymer: mRNA ratio vaccine ink. mRNA recovery increased as polymermRNA ratio increased, and mRNA encapsulation efficiency was the highest in vaccine ink composed of LNPs of NP ratio 5.4. (FIG. 18F) Histogram of mRNA integrity analysis obtained by capillary gel electrophoresis after re-dissolution.

FIGs. 19A-19F show the structure of mRNA-LNPs upon redissolution in representative cryo-TEM and corresponding FTT for single LNPs following redissolution from the PVPPVA polymer matrix.

FIGs. 20A-20C show in vivo imaging studies with Flue mRNA-LNPs loaded in MAPs at different polymer: mRNA ratios of 32, 160, 320, 640 and 960. MAPs were applied to BALB/c mice at mRNA dose of 1 pg. FIG. 20A shows images of mice showing in vivo bioluminescence analyzed 24 h after injection. FIGs. 20B & 20C show quantification of the biohiminescent intensity of photons emitted from each MAP in image A. Statistical significance was calculated by Student t-test: *P < 0.05 and **P < 0.01. Data are shown as means ± SEM n=4.

FIGs. 21A-21J show mRNA MAP drying cycle reduction and application in vivo. FIG. 21A shows a schematic depicting MAP with and without polymer solution backing, resulting in either one or two drying cycles. Drying cycles do not significantly affect mRNA-LNP diameter (n = 1-5 independent samples) (FIG. 21B) but decrease mRNA integrity (n = 5 independent samples) (C) Ordinary two-way ANOVA (Sidak’s multiple comparisons test). (FIG. 21C) Loading of encapsulated mRNA in microneedles is not significantly changed by ink concentration or multiple drying cycles through polymer backing application (n = 5 independent samples). Ordinary two- way ANOVA (Sidak’s multiple comparisons test) (FIGs. 21D & 21E) 3X ink concentration is required to fully fill mechanically stable microneedles without the addition of a polymer backing. (FIG. 21F) Removing the additional drying step results in MAPs with greater mRNA expression in BALB/c mice (n = 10 biologically independent samples). Non-parametric one-way ANOVA (Kruskal-Wallis multiple comparisons test). (FIG. 21G) MAPs loaded with mRNA-LNPs expressing the SARS-CoV-2 S protein receptor binding domain (RBD) were used to immunize BALB/c mice with footpad application and compared against intramuscular (IM) administration from 0 to 10 pg. An electrochemiluminescent assay for serum anti -RBD binding responses was used. (FIG. 21H) MAP size and mRNA loading were increased by depositing more vaccine ink in a larger microneedle mold. MAPs were used to immunize Wistar rats and serum was analyzed by ELISA for anti-RBD binding titers (FIG. 211) and pseudovirus neutralizing antibody titers (FIG. 21J). *P < 0.05; **P < 0.01; ***P < 0.001; ****p < 0.0001. Data represent means ± s.d.

FIG. 22 representative cryogenic transmission electron microscopy (cryo-TEM) images of mRNA LNPs of NP ratios of 1.5, 3.1, and 5.4 made with unlabeled or Au-labeled RNA (constituting 1/3 of the total mRNA) showing the distribution of Au-RNA within LNPs.

FIGs. 23A shows the DLS analysis of mRNA-LNPs of NP ratios of 1.5 and 3.1 in solution and after vacuum-drying.

FIG. 23B shows that empty microneedles show no spherical morphologies resembling LNPs.

FIG. 23C shows individual microneedles were sectioned near the tip of the microneedle.

FIG. 23D shows LNP size determined by analyzing SEM images of MAP cross sections (n = 17-467 independent observations over three images). FIG. 23E shows the theoretical percentage of an SEM cross section occupied by mRNA- LNPs decreases as polymer: mRNA increases. However, the actual coverage based on the number and size of LNPs observed in the SEM cross section may not reach the theoretical value due to stability issues, such as cholesterol crystallization, bleb formation, or aggregation into large, nonspherical morphologies. At high polymer: mRNA ratios, the two converge, indicating that dried LNP integrity is better preserved.

FIGs 24A-24D show analysis of mRNA-LNPs as vaccine ink dries. FIG. 24A shows the thermogravimetric analysis was used to determine the water content in samples of PVPVA ink containing mRNA-LNPs over 24 hours. Encapsulation efficiency was also assessed at each time point. FIG. 24B shows alternative drying materials such as sucrose/maltose buffer and PBS alone were added to the drying time assessment of encapsulation efficiency. FIGs. 24C & 24D show recovery efficiency (FIG. 24C) and LNP diameter (FIG. 24D) were also assessed at each time point.

FIGs. 25A & 25B show the effect of surface-area-to-volume ratio of vaccine device on mRNA-LNPs. mRNA-LNP devices were fabricated with different sizes in the bottom of a test tube. Surface area and volume were estimated based on the dimensions of the test tube. The device was redissolved and mRNA-LNP (FIG. 25A) encapsulation efficiency and (FIG. 25B) diameter were analyzed.

FIG. 26 shows mRNA encapsulation efficacy (EE) was measured by ribogreen assay. Effect of various polymer: mRNA ratios on mRNA recovery and free mRNA.

FIGs. 27A-27G show mRNA-LNPs formulations containing PVPVA that were compared against sucrose/maltose buffer for vacuum drying and lyophilizing processes. Devices were initially redissolved immediately after fabrication, and mRNA-LNPs were assessed for (FIG. 27A) recovery efficiency, (FIG. 27B) encapsulation efficiency, (FIG. 27C) diameter, and (FIG. 27D) luminescence 24 h after i.m. administration of 1 pg of total FLuc mRNA in BALB/c mice. Then, devices were stored for one month at -20°C, 4°C, and 25°C and assessed for (FIG. 27E) diameter, (FIG. 27F) encapsulation efficiency, and (FIG. 27G) luminescence 24 h after i.m. administration of 1 pg of encapsulated FLuc mRNA in BALB/c mice. Some formulations could not be injected due to high volume of redissolution buffer needed for injection.

FIG. 28 shows that concentrated ink yields one cycle MAPs with improved mechanical properties. No backing MAPs fabricated using a concentrated 3X vaccine ink have comparable stiffness to IX MAPs with a PVPVA backing. As expected, stiffness increases when a backing is applied.

FIG. 29 shows the immunogenicity of MAPs in comparison to ID and IM controls. MAPs loaded with mRNA-LNPs expressing the SARS-CoV-2 S protein receptor binding domain (RBD) were used to immunize BALB/c mice with footpad application and compared against intradermal (ID) and intramuscular (IM) administration from 0 to 10 pg. An electrochemiluminescent assay for serum anti-RBD binding responses was used.

DETAILED DESCRIPTION OF THE INVENTION

In certain aspects, disclosed herein are injectable needles for administering vaccine and other therapeutic agents that require multiple doses (e.g., vaccine boosters). The system uses ‘SEAL’ (StampEd Assembly of polymer Layers) technology to produce core-shell microparticles (MP), with tuneable release kinetics based on the materials used. SEAL technology allows for any antigen to be encapsulated into biodegradable polymer particles that have delayed pulsatile releases at predetermined boost points. By engineering the shell material (which is made of PLGA (Poly(lactic-co-glycolic acid)) with different molecular weights), we one can control when the cargo gets released. By encapsulating vaccine doses into microparticles with different PLGA shells and administering all of them with a single injection, we can deliver multiple vaccine doses with one administration. Therefore, patients do not need to go back to clinic for second or third booster shot.

Here, we are embedding a SEAL particle in a readily dissolvable polymer needle (made of PVP-PVA mixture (Polyvinylpyrrolidone (PVP)) (Poly(vinyl alcohol) (PVA))) to deliver enough mRNA vaccine dose (in a microgram range) without necessitating hypodermic metal needles. First, we make elongated core-shell (CS) SEAL particle (450um x 450um x 4mm), we loaded vaccine in the core and in SEAL the particle. Then, we load the SEAL particle into a needle-shaped PDMS (Polydimethylsiloxane) negative mold, and dispense polymer mixture (PVP-PVA) on top of the mold. This polymer mixture has mRNA vaccine mixed in it as well. We call this vaccine ink (mRNA-LNP + PVP + PVA). This formed the outer needle and will dissolve within 10-15 minutes once administered, and deliver the 1st vaccine dose (prime dose). Because PDMS is air permeable but not liquid permeable, when we applied vacuum directly under the PDMS mold, it sucked out the air pocket in the mold, and filled the cavity with the vaccine ink, thus allowing it to be molded into a needle shape. We dried the needle in the mold. Once it was demolded, it was now a solid dissolvable needle that contains 1 st dose (prime dose) in the dissolvable needle part and 2nd dose (booster dose) in the CS MP (core-shell microparticle). The needle was designed to be retractable in case the patient has an adverse reaction to the vaccine (e.g., and allergic reaction).

The readily dissolvable needle contains the prime dose, so it can dissolve and deliver prime dose within minutes. The SEAL macroparticle inside the needle contains the booster dose, and its release timepoint can be programmed by changing the particle material. And the entire “microparticle-embedded dissolvable needle” can be injected subcutaneously using a spring applicator. This approach offers many advantages, such as: delivering two mRNA vaccine doses in a single shot, a large core space for containing a large mRNA vaccine dose, tunable design for different applications. Because this entire needle is intact in a dry state, we can ensure injectability without any residue and more accurate release kinetics compared to when administering microparticles with solvent, as our design experiences no shear stress or early exposure to fluid. This dryness also offers thermostability. mRNA-LNP vaccine has shown excellent stability/potency even at room temperature when solidified in a PVP-PVA matrix, meaning there is no need for cold-chain (sub-zero infrastructure during transportation or storage). Because we are injecting the whole dissolvable needle without needing extra hypodermic metal needle, it makes the administration simpler and allows administration with smaller needle sizes (21-25G), which is less painful and adequate for children. Critically, the injectable needles also create no sharps waste.

The injectable needle may comprise a plurality of compartments (e.g., 2, 3, 4, or more compartments), wherein each compartment encapsulates a composition, and each compartment releases its encapsulated composition at a different point in time. Each encapsulated composition may be the same as or different to the others.

In one aspect, the present disclosure provides injectable needles comprising a needle body, a needle tip, a compartment, a first composition, and a second composition; wherein: the needle body and the needle tip are connected to one another; the first composition and the compartment are encapsulated by the needle tip and the needle body; the compartment comprises a compartment base and a compartment cap; and the second composition is encapsulated in the compartment. In certain embodiments, the length of the injectable needle is 1 - 10 mm. In certain embodiments, the length of the injectable needle is about 1 mm, about 2 mm, about 3 mm, about 4 mm, about 5 mm, or about 6 mm. In certain embodiments, the length of the injectable needle is about 4 mm, about 5 mm, or about 6 mm.

In certain embodiments, the width of the injectable needle is 100 - 1,500 gm. In certain embodiments, the width of the injectable needle is about 200 gm, about 300 gm, about 400 gm, about 500 gm, about 600 gm, about 700 gm, about 800 gm, about 900 gm, about 1,000 gm, about 1,100 gm, about 1,200 gm, about 1,300 gm, about 1,400 gm, or about 1,500 gm. In certain embodiments, the width of the injectable needle is about 450 gm.

In certain embodiments, the height of the injectable needle is 100 - 1,000 gm. In certain embodiments, the height of the injectable needle is about 200 gm, about 300 gm, about 400 gm, about 500 gm, or about 600 gm. In certain embodiments, the height of the injectable needle is about 450 gm.

In certain embodiments, the injectable needle is a 16 gauge, 18 gauge, 19 gauge, 20 gauge, 21 gauge, 22 gauge, 23 gauge needle, 28 gauge, or 30 gauge needle. In certain embodiments, the injectable needle is a 21 gauge, 22 gauge, or 23 gauge needle.

In certain embodiments, the needle body comprises a polymer or a copolymer. In certain embodiments, the needle body comprises a blend of polymers (e.g., a blend of 2, 3, 4, or 5 polymers, preferably a blend of 2 polymers).

In certain embodiments, the needle body comprises a blend of polyvinylpyrrolidone (PVP) and polyvinylalcohol (PVA), or a blend of PVP, PVA, and sucrose; preferably a blend of PVP and PVA. In certain embodiments, the PVP is PVP10 (i.e., PVP with an average molecular weight of 10 kDa) or PVP60 (i.e., PVP with an average molecular weight of 60 kDa). In certain embodiments the ratio of PVP:PVA in the polymer blend is 1: 1, 2: 1, or 5: 1. In certain preferred embodiments, the needle body comprises a copolymer comprising PVP60 and PVA at a ratio of 1: 1. In certain embodiments, the needle body comprises a random copolymer. In certain embodiments, the needle body comprises a block copolymer.

In certain embodiments, the needle tip comprises a polymer or a copolymer. In certain embodiments, the needle tip comprises a blend of polymers (e.g., a blend of 2, 3, 4, or 5 polymers, preferably a blend of 2 polymers). In certain embodiments, the needle tip comprises a blend of PVP and PVA or a blend of PVP, PVA, and sucrose, preferably a blend of PVP and PVA. In certain embodiments, the PVP is PVP10 (i.e., PVP with an average molecular weight of 10 kDa) or PVP60 (i.e., PVP with an average molecular weight of 60 kDa). In certain preferred embodiments, the PVP is PVP 10. In other preferred embodiments, the PVP is PVP60. In certain embodiments, the ratio of PVP:PVA in the polymer blend is 1: 1, 2: 1, or 5:1. In certain embodiments, the w/w ratio of PVP:PVA in the polymer blend is 1: 1, 2: 1, or 5: 1. In certain preferred embodiments, the needle tip comprises a blend of PVP60 and PVA at a w/w ratio of 5: 1. In certain preferred embodiments, the needle tip comprises a blend of PVP 10 and PVA at a w/w ratio of 1:1. In certain embodiments, the needle tip comprises a block copolymer. In certain embodiments, the needle tip comprises a random copolymer. In certain preferred embodiments, the first composition is encapsulated in the needle tip.

In certain embodiments, the compartment base comprises a polymer. In certain embodiments, the compartment base comprises a copolymer. In certain embodiments, the compartment base comprises poly lactic-co-glycolic acid (PLGA). In certain preferred embodiments, the compartment base comprises PLGA 858 (i.e., a copolymer of lactide: glycolide at a w/w ratio of 85:15). In certain embodiments, the compartment base comprises a random copolymer. In certain embodiments, the compartment base comprises a block copolymer.

In certain embodiments, the compartment cap comprises a polymer. In certain embodiments, the compartment cap comprises a copolymer. In certain preferred embodiments, the compartment cap comprises PLGA 502 (i.e., a copolymer of lactide: glycolide at a w/w ratio of 1: 1). In certain embodiments, the compartment cap comprises a random copolymer. In certain embodiments, the compartment cap comprises a block copolymer.

In certain embodiments, the first composition or the second composition comprises a nanoparticle (e.g., a polymeric nanoparticle of a gold nanoparticle or a lipid nanoparticle). In certain embodiments, the first composition comprises a nanoparticle (e.g., a polymeric nanoparticle of a gold nanoparticle or a lipid nanoparticle). In certain embodiments, the second composition comprises a nanoparticle (e.g., a polymeric nanoparticle of a gold nanoparticle or a lipid nanoparticle). In certain embodiments, the first composition or the second composition comprises a lipid nanoparticle comprising an ionizable lipid, a PEG lipid, a phospholipid, or a combination of the foregoing. In certain embodiments, the first composition comprises a lipid nanoparticle comprising an ionizable lipid, a PEG lipid, a phospholipid, or a combination of the foregoing. In certain embodiments, the second composition comprises a lipid nanoparticle comprising an ionizable lipid, a PEG lipid, a phospholipid, or a combination of the foregoing.

In certain preferred embodiments, the first composition or the second composition comprises: i) a polymer or a copolymer and a lipid nanoparticle, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is at least 100: 1; or ii) a polymer or a copolymer and a bioactive agent, wherein the w/w ratio of the polymer or the copolymer to the bioactive agent is at least 1000: 1.

In certain embodiments, the first composition or the second composition comprises a polymer or a copolymer and a lipid nanoparticle, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is at least 100: 1. In certain embodiments, the first composition comprises a polymer or a copolymer and a lipid nanoparticle, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is at least 100: 1. In certain embodiments, the second composition comprises a polymer or a copolymer and a lipid nanoparticle, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is at least 100: 1.

In certain embodiments, the second composition comprises a polymer or a copolymer and a lipid nanoparticle, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is at least 100: 1. In certain embodiments, the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is about 100: 1, 150: 1, 200:1, 250:1, 300: 1, 350: 1, 400: 1, 450: 1, or 500: 1. In certain embodiments, the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is about 333:1.

In certain embodiments, the lipid nanoparticle comprises an ionizable lipid, cholesterol, a polyethyleneglcyol lipid, a phospholipid, or a combination thereof. In certain embodiments, the lipid nanoparticle comprises 1 -octylnonyl 8-[(2-hydroxyethyl)[8-(nonyloxy)-8- oxooctyl] amino] octanoate (i.e., lipid 5, CAS Ref. No.: 2089251-33-0). In certain embodiments, the lipid nanoparticle consists essentially of 1-octylnonyl 8-[(2-hydroxyethyl)[8-(nonyloxy)-8- oxooctyl] amino] octanoate (i.e., lipid 5). In certain embodiments, the lipid nanoparticle comprises 3,6-bis(4-(bis(2-hydroxydodecyl)amino)butyl)piperazine-2,5-dione (i.e., cKK-E12). In certain embodiments, the lipid nanoparticle consists essentially of 3,6-bis(4-(bis(2- hydroxydodecyl)amino)butyl)piperazine-2, 5-dione (i.e., cKK-E12). In certain embodiments, the diameter of the lipid nanoparticle is about 50 nm, about 75 nm, about 100 nm, about 125 nm, about 150 nm, about 175 nm, about 200 nm, about 225 nm, about 250, about 275 nm, or about 300 nm. In certain embodiments, the diameter of the lipid nanoparticle is about 125 nm. In certain embodiments, the diameter of the lipid nanoparticle is about 225 nm.

In certain preferred embodiments, the injectable needed further comprises a bioactive agent.

In certain embodiments, the w/w ratio of the polymer or the copolymer to the bioactive agent is at least 1000: 1. In certain embodiments, the w/w ratio of the polymer or the copolymer to the bioactive agent is about 600: 1, about 700: 1, about 800: 1, 900: 1, or about 1000: 1. In certain embodiments, the w/w ratio of the polymer or the copolymer to the bioactive agent is about 1,250: 1, about 1500: 1, about 1750: 1, about 2000:1, about 2,250:1, about 2,500: 1, about 2,750: 1, or about 3000: 1. In certain embodiments, the w/w ratio of the polymer or the copolymer to the bioactive agent is about 4,000: 1, about 5,000: 1, about 6,000: 1, about 7000: 1, about 8,000: 1, about 9,000: 1, or about 10,000: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is about 20: 1, about 40:1, about 60: 1, about 80: 1, about 100: 1, 120: 1, 140:1, 160:1, 180: 1, 200: 1, 220: 1, 240: 1, 260:1, 280: 1, or about 300:1, about 320:1, about 340:1, about 360:1, about 380: 1, or about 400:1. In certain preferred embodiments, the w/w ratio of copolymer to bioactive agent is about 50: 1. In certain preferred embodiments, the w/w ratio of copolymer to bioactive agent is about 160:1. In certain preferred certain embodiments, the w/w ratio of copolymer to bioactive agent is about 320: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 100: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 150: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 200: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 250: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 300: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 320: 1.

In certain embodiments, the first composition or the second composition further comprises a polymer. In certain embodiments, the first composition further comprises a polymer. In certain embodiments, the second composition further comprises a polymer. In certain embodiments, the polymer is PVP. In certain embodiments, the PVP is PVP10. In certain embodiments, the PVP is PVP60. In certain embodiments, the polymer is polyvinylalcohol.

In certain embodiments, the first composition or the second composition further comprises a copolymer. In certain embodiments, the first composition further comprises a copolymer. In certain embodiments, the second composition further comprises a copolymer. In certain embodiments, the copolymer comprises a plurality of repeat units of vinylalcohol; a plurality of repeat units of vinylpyrrolidinone; and the copolymer is a block copolymer or a random copolymer. In certain embodiments, the copolymer consists essentially of a plurality of repeat units of vinylalcohol; and a plurality of repeat units of vinylpyrrolidinone; and the copolymer is a block copolymer or a random copolymer. In certain embodiments, the mass ratio of polyvinylalcohol to polyvinylpyrrolidone is about 1 :1, about 2:1, about 3:1, about 4:1, about 5:1, or about 6: 1. In certain embodiments, the mass ratio of polyvinylalcohol to polyvinylpyrrolidone is about 1: 1, about 2: 1, or about 3:1. In certain preferred embodiments, the mass ratio of polyvinylalcohol to polyvinylpyrrolidone is about 1: 1. In certain preferred embodiments, the mass ratio of polyvinylalcohol to polyvinylpyrrolidone is about 2:1. In certain embodiments, the copolymer comprises a plurality of repeat units of vinylalcohol; a plurality of repeat units of sucrose; and the copolymer is a block copolymer or a random copolymer. In certain embodiments, the copolymer consists essentially of a plurality of repeat units of vinylalcohol; a plurality of repeat units of sucrose; and the copolymer is a block copolymer or a random copolymer. In certain embodiments, the mass ratio of polyvinylalcohol to sucrose is about 1: 1, about 2: 1, about 3:1, about 4:1, about 5: 1, or about 6: 1. In certain embodiments, the mass ratio of polyvinylalcohol to sucrose is about 1: 1 or about 2: 1. In certain embodiments, the copolymer comprises a plurality of repeat units of vinylalcohol, a plurality of repeat units of polyvinylpyrrolidone, a plurality of repeat units of sucrose; and the copolymer is a block copolymer or a random copolymer. In certain embodiments, the copolymer consists essentially of a plurality of repeat units derived from vinylalcohol, a plurality of repeat units derived from polyvinylpyrrolidone, a plurality of repeat units derived from sucrose; and the copolymer is a block copolymer or a random copolymer. In certain embodiments, the mass ratio of polyvinylalcohol to polyvinylpyrrolidone to sucrose is about 1: 1: 1, about 1: 1:2, or about 1 : 1 :3. In certain embodiments, the mass ratio of polyvinylalcohol to polyvinylpyrrolidone to sucrose is about 1: 1:2. In certain embodiments, the copolymer is a block copolymer. In certain embodiments, the copolymer is a random copolymer.

In certain embodiments, the polymer comprises 50 - 500 repeat units. In certain embodiments, the polymer comprises 50 - 250 repeat units. In certain embodiments, the polymer comprises 75 - 125 repeat units. In certain embodiments, the polymer comprises about 70, about 80, about 90, about 100, about 110, or about 120 repeat units. In certain embodiments, the polymer comprises about 90 repeat units.

In certain embodiments, the copolymer comprises 250 - 1,500 repeat units. In certain embodiments, the copolymer comprises 500 - 1250 repeat units. In certain embodiments, the copolymer comprises 500 - 1000 repeat units. In certain embodiments, the copolymer comprises 600 - 800 repeat units. In certain embodiments, the copolymer comprises about 600, about 650, about 700, about 750, about 800, about 850, or about 900 repeat units. In certain embodiments, the copolymer comprises about 700 repeat units.

In certain embodiments, the first composition or the second composition further comprises a bioactive agent. In certain embodiments, the first composition further comprises a bioactive agent. In certain embodiments, the second composition further comprises a bioactive agent. In certain preferred embodiments, the first composition and the second composition further comprise a bioactive agent.

In certain embodiments, the concentration of the bioactive agent is about 50 pg/mL, about 100 pg/mL, about 150 pg/mL, about 200 pg/mL, about 250 pg/mL, about 300 pg/mL, about 350 pg/mL, about 400 pg/mL, about 450 pg/mL, about 500 pg/mL, about 550 pg/mL, about 600 pg/mL, about 650 pg/mL about 700 pg/mL, about 750 pg/mL, about 800 pg/mL, about 850 pg/mL, about 900 pg/mL, about 950 pg/mL or about 1,000 pg/mL. In certain embodiments, the concentration of the bioactive agent is about 650 pg/mL, about 700 pg/mL, or about 750 pg/mL. In certain embodiments, the concentration of the bioactive agent is about 700 pg/mL. In certain embodiments, the weight of the bioactive agent is about 2 pg, about 4 pg, about 6 pg, about 8 pg, about 10 pg, about 12 pg, or about 14 pg, about 16 pg. In certain preferred embodiments, the weight of the bioactive agent is about 6 pg. In certain preferred embodiments, the weight of the bioactive agent is about 7 pg.

In certain embodiments, the w/w ratio of copolymer to bioactive agent is about 20: 1 , about 40:1, about 60: 1, about 80: 1, about 100: 1, 120: 1, 140:1, 160: 1, 180: 1, 200:1, 220:1, 240: 1, 260: 1, 280: 1, or about 300: 1, about 320:1, about 340: 1, about 360: 1, about 380: 1, or about 400: 1. In certain preferred embodiments, the w/w ratio of copolymer to bioactive agent is about 50: 1. In certain preferred embodiments, the w/w ratio of copolymer to bioactive agent is about 160: 1. In certain preferred embodiments, the w/w ratio of copolymer to bioactive agent is about 320: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 100: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 150: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 200: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 250: 1. In certain embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 300: 1. In certain preferred embodiments, the w/w ratio of copolymer to bioactive agent is greater than about 320: 1.

In certain embodiments, the nitrogen to phosphate ratio of the lipid nanoparticle is about 1.5. In certain embodiments, the nitrogen to phosphate ratio of the lipid nanoparticle is about 3.1. In certain embodiments, the nitrogen to phosphate ratio of the lipid nanoparticle is about 5.4

In certain embodiments, the encapsulation efficiency of the lipid nanoparticle is greater than about 40%, about 50%, about 60%, about 70%, about 80%, or about 90%. In certain embodiments, the encapsulation efficiency of the lipid nanoparticle is greater than about 60%.

In certain embodiments, the bioactive agent defined in the compositions herein has improved stability (e.g., improved thermostability) as compared to the bioactive agent alone or the bioactive agent in a composition not defined in the embodiments recited herein.

In certain embodiments, each bioactive agent is independently selected from the group consisting of a protein, an antibody, a nucleic acid, a small molecule, and a vaccine. In certain embodiments, at least one bioactive agent is a mRNA, siRNA, RNA, or DNA. In certain embodiments, the at least one bioactive agent is an antibody. In certain embodiments, the at least one bioactive agent is a small molecule (e.g., a drug). In certain embodiments, the at least one bioactive agent is a protein. In certain preferred embodiments, the at least one bioactive agent is a vaccine. In certain further preferred embodiments, the at least one bioactive agent is a nucleic acid. In certain very preferred embodiments, the at least one bioactive agent is an RNA (e.g., an mRNA).

In certain embodiments, the first composition or the second composition further comprises a pharmaceutically acceptable carrier. In certain embodiments, the first composition or the second composition further comprises an adjuvant. In certain embodiments, the injectable needle is dissolvable. In certain embodiments, the injectable needle has been formed using a single drying step.

In certain embodiments: the needle body comprises a copolymer comprising PVP60 and PVA at a w/w ratio of 1 : 1 ; the needle tip comprises a blend of PVP10 and PVA at a w/w ratio of 1 : 1; the compartment base comprises PLGA 858 (i.e., a copolymer of lactide: glycolide at a ratio of w/w 85: 15); the compartment cap comprises PLGA 502 (i.e., a copolymer of lactide: glycolide at a ratio of 1: 1); the first composition comprises:

PVP 10 and PVA at a w/w ratio of 1 : 1 ; a bioactive agent further wherein: the weight of the bioactive agent is about 7 pg; and the w/w ratio of copolymer to bioactive agent is about 320: 1; the second composition comprises:

PVP 10 and PVA at a w/w ratio of 1 :2; a bioactive agent further wherein: the weight of the bioactive agent is about 6 pg; and the w/w ratio of copolymer to bioactive agent is about 50: 1.

In another aspect, the present disclosure provides methods of delivering a therapy to a subject in need thereof comprising contacting the subject with an injectable needle disclosed herein. In certain embodiments, the therapy is a vaccine. In certain embodiments, the therapy is an mRNA vaccine. In certain embodiments, the therapy is an anticancer therapy, an antibacterial therapy, or an anticancer therapy. In certain embodiments, the therapy is delivered in the form of a first dose and a second dose. In certain embodiments, the first dose and the second dose are delivered 1 day to six months apart. In certain embodiments, the first dose and the second dose are delivered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 days apart. In certain embodiments, the therapy is delivered in the form of a plurality of doses (e.g., 2, 3, 4, or more doses). Certain single-injection and pulsatile-release microdevices are disclosed in US 10,300,136, US 10,960,073, US 2021/0205444 Al, and US 2019/0076631 Al, the contents of each of which are incorporated by reference herein. Related LNPs are disclosed in WO 2023/038892, the contents of which are fully incorporated by reference herein.

Pharmaceutical Compositions

The compositions and methods of the present invention may be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or the compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In preferred embodiments, when such pharmaceutical compositions are for human administration, particularly for invasive routes of administration (i.e., routes, such as injection or implantation, that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipients can be chosen, for example, to effect delayed release of an agent or to selectively target one or more cells, tissues or organs.

A pharmaceutically acceptable carrier can contain physiologically acceptable agents that act, for example, to stabilize, increase solubility or to increase the absorption of a compound such as a compound of the invention. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a self-emulsifying drug delivery system or a self-micro- emulsifying drug delivery system. The pharmaceutical composition (preparation) also can be a liposome or other polymer matrix, which can have incorporated therein, for example, a compound of the invention. Liposomes, for example, which comprise phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about ninety-nine percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

For use in the methods of this invention, active compounds can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier. Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound(s) being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the pharmaceutical composition or compound at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. By “therapeutically effective amount” is meant the concentration of a compound that is sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the weight, sex, age, and medical history of the subject. Other factors which influence the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound of the invention. A larger total dose can be delivered by multiple administrations of the agent. Methods to determine efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison’s Principles of Internal Medicine 13 ed., 1814-1882, herein incorporated by reference).

In general, a suitable daily dose of an active compound used in the compositions and methods of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above. The patient receiving this treatment is any animal in need, including primates, in particular humans; and other mammals such as equines, cattle, swine, sheep, cats, and dogs; poultry; and pets in general.

In certain embodiments, compounds of the invention may be used alone or conjointly administered with another type of therapeutic agent.

The present disclosure includes the use of pharmaceutically acceptable salts of compounds of the invention in the compositions and methods of the present invention. In certain embodiments, contemplated salts of the invention include, but are not limited to, alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, L-arginine, benenthamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, IH-imidazole, lithium, L-lysine, magnesium, 4-(2- hydroxyethyljmorpholine, piperazine, potassium, 1 -(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, 1 -hydroxy -2- naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4- acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, 1-ascorbic acid, 1-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane- 1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, 1-malic acid, malonic acid, mandelic acid, methanesulfonic acid , naphthalene-l,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, proprionic acid, 1-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, 1-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid acid salts.

The pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent.

Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alphatocopherol, and the like; and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Definitions

Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art. Generally, nomenclature used in connection with, and techniques of, chemistry, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, pharmacology, genetics and protein and nucleic acid chemistry, described herein, are those well known and commonly used in the art.

The methods and techniques of the present disclosure are generally performed, unless otherwise indicated, according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout this specification. Chemistry terms used herein, unless otherwise defined herein, are used according to conventional usage in the art, as exemplified by “The McGraw-Hill Dictionary of Chemical Terms”, Parker S., Ed., McGraw-Hill, San Francisco, C.A. (1985).

All of the above, and any other publications, patents and published patent applications referred to in this application are specifically incorporated by reference herein. In case of conflict, the present specification, including its specific definitions, will control.

The term “agent” is used herein to denote a chemical compound (such as an organic or inorganic compound, a mixture of chemical compounds), a biological macromolecule (such as a nucleic acid, an antibody, including parts thereof as well as humanized, chimeric and human antibodies and monoclonal antibodies, a protein or portion thereof, e.g., a peptide, a lipid, a carbohydrate), or an extract made from biological materials such as bacteria, plants, fungi, or animal (particularly mammalian) cells or tissues. Agents include, for example, agents whose structure is known, and those whose structure is not known.

A “patient,” “subject,” or “individual” are used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (including bovines, porcines, etc.), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats).

“Treating” a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.

The term “preventing” is art-recognized, and when used in relation to a condition, such as a local recurrence (e.g., pain), a disease such as cancer, a syndrome complex such as heart failure or any other medical condition, is well understood in the art, and includes administration of a composition which reduces the frequency of, or delays the onset of, symptoms of a medical condition in a subject relative to a subject which does not receive the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment relative to an untreated control population, and/or delaying the appearance of detectable cancerous growths in a treated population versus an untreated control population, e.g., by a statistically and/or clinically significant amount.

“Administering” or “administration of’ a substance, a composition or an agent to a subject can be carried out using one of a variety of methods known to those skilled in the art. For example, a composition or an agent can be administered, intravenously, arterially, intradermally, intramuscularly, intraperitoneally, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (by absorption, e.g., through a skin duct). A composition or agent can also appropriately be introduced by rechargeable or biodegradable polymeric devices or other devices, e.g., patches and pumps, or formulations, which provide for the extended, slow or controlled release of the composition or agent. Administering can also be performed, for example, once, a plurality of times, and/or over one or more extended periods.

Appropriate methods of administering a substance, a composition or an agent to a subject will also depend, for example, on the age and/or the physical condition of the subject and the chemical and biological properties of the composition or agent (e.g., solubility, digestibility, bioavailability, stability and toxicity). In some embodiments, a composition or an agent is administered orally, e.g., to a subject by ingestion. In some embodiments, the orally administered composition or agent is in an extended release or slow release formulation, or administered using a device for such slow or extended release.

As used herein, the phrase “conjoint administration” refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered therapeutic agent is still effective in the body (e.g., the two agents are simultaneously effective in the patient, which may include synergistic effects of the two agents). For example, the different therapeutic compositions can be administered either in the same formulation or in separate formulations, either concomitantly or sequentially. Thus, an individual who receives such treatment can benefit from a combined effect of different therapeutic agents.

A “therapeutically effective amount” or a “therapeutically effective dose” of a drug or agent is an amount of a drug or an agent that, when administered to a subject will have the intended therapeutic effect. The full therapeutic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. The precise effective amount needed for a subject will depend upon, for example, the subject’s size, health and age, and the nature and extent of the condition being treated, such as cancer or MDS. The skilled worker can readily determine the effective amount for a given situation by routine experimentation.

As used herein, the terms “optional” or “optionally” mean that the subsequently described event or circumstance may occur or may not occur, and that the description includes instances where the event or circumstance occurs as well as instances in which it does not.

The phrase “pharmaceutically acceptable” is art-recognized. In certain embodiments, the term includes compositions, excipients, adjuvants, polymers and other materials and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The phrase “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filter, diluent, excipient, solvent or encapsulating material useful for formulating a drug for medicinal or therapeutic use.

EXAMPLES

The invention now being generally described, it will be more readily understood by reference to the following examples which are included merely for purposes of illustration of certain aspects and embodiments of the present invention and are not intended to limit the invention.

Ex ample 1: Preparation of Exemplary Lipid Nanoparticles

Lipids were dissolved in ethanol at a molar ratio of 35: 16:46.5:2.5 (cKK-E12 (Organix):DOPE (Avanti): Cholesterol (Sigma):C14-PEG2000 (Avanti)) or 50: 10:38.5: 1.5 (Lipid 5(Organix):DOPE:Cholestorol:C14-PEG2000) when using respectively cKK-E12 or Lipid 5 as the ionizable lipid. To prepare the LNPs, the ethanoic solution was rapidly added to and pipette- mixed with a mRNA solution buffered with citrate at pH 3 at volume ratio 3:1 (aqueous: ethanol). The ionizable lipid to mRNA weight ratio was set to 10 and the final mRNA concentration was 0.1 mg mL-1. All nucleic acids were stored at - 80 °C and were allowed to thaw on ice prior to use. The LNPs were then dialyzed for at least 2 hours in PBS at 4°C in a 20,000 MWCO cassette.7 Lor LNPs in deionized (DI) water, the solution was dialyzed against DI water for an additional minimum of 2 hours at 4°C. When needed, the LNPs were concentrated on an Amicon filter by centrifugation at 3000 x g.19 All solutions were kept at 4°C and used within a week. mRNA concentration and encapsulation efficiency in the LNPs was estimated using a Quant- iT RiboGreen assay (ThermoLisher) and a modified procedure described elsewhere.7 Briefly, LNPs were diluted in either Tris-EDTA (IE) or IE mixed with triton X-100 buffer (TX). Then, the procedure recommended by the manufacturer was used to quantify the mRNA that is not encapsulated (when diluted with TE) and the total mRNA concentration (when diluted with TX). Lor size and surface potential, LNPs were diluted 200 times in PBS and measurement was achieved using [REP Malvern], When measuring the mRNA loading in MNPs, needles were cut, and dissolved in TE and TX. The total mRNA concentration found was used to estimate the encapsulated mRNA concentration based on its encapsulation efficiency.

Example 2: Preparation of Exemplary Polymer;Lipid Nanoparticle formulations

Polymers were solubilized in PBS or DI water at a concentration ranging from 10% to 30% w/w depending on their solubility. These solutions were then weighted and mixed with LNPs solution to reach the appropriate polymer to mRNA mass ratio. The mixture was immediately dried in a Low-Bind Eppendorf tube in a desiccator under -0.5 bar vacuum. After 24 hours drying, the pellet was redissolved inPBS and incubatedfor 10min at 37°C. The PBS volume used was adjusted so that 15 pL of solution contains 50 ng of encapsulated mRNA. This solution was used to transfect cells.

Example 3: Quantification of Properties of Exemplary Polvmer:Lipid Nanoparticle formulations

HeLa cells were cultured in high glucose Dulbecco’s Modified Eagles Medium with phenol red (DMEM, Invitrogen) supplemented with 10% FBS (Invitrogen) and 1% antibiotic (Invitrogen). 10,000 cells were seeded in wells of a white 96-well plate in full growth medium. 20 hours after seeding, 15 pL of fresh LNPs or dissolved formulation were added to the growth medium. In all cases, 50 ng of encapsulated mRNA were added to each well. 24 hours after transfection, 100 pL of Bright-Glo Luciferase Assay Kit (Promega) reagent was added and luminescence was measured in the following 2 minutes using a Tecan multiplate reader.

Example 4: Fabrication of an exemplary injectable needle

The CS MP fabrication method is based on SEAL MP fabrication that was previously reported. Science, 2017, 357(6356), 1138-42. However, we enlarged the MP dimensions to deliver effective dose of mRNA vaccine. We then made the PLGA shell bases and shell caps separately. For the cargo, we made a “mRNA vaccine block” that is composed of LNP-encapsulated mRNA, polymer mixture (e.g., PVP-PVA), and water. We dispense mRNA vaccine ink first to a negative PDMS mold, use vacuum-thru technique to mold the ink to block shapes, dry overnight, demold, and cut individual vaccine blocks. This method improves mRNA-LNP stability, deliverable mass, reproducibility, scalability (e.g., we can make the mold larger and make hundreds of thousands of vaccine blocks using the same technique and the same amount of time), while reducing labor. Once we have the shell base, shell cap, and a dry vaccine block, we load the block into the shell base. Then, SEAL the base and cap together and cut the sealed microparticle. The CS MP is now ready. In a negative PDMS needle mold, we first inject 1st vaccine dose into the needle tip part and then load the CS MP which contains the 2nd dose. Let it dry for a few hours, and then load the rest of the needle body material. Let the whole needle dry for a few days, and once you demold the needle, we have the injectable needle. We use this 2-step loading vacuum-thru technique to fabricate our needle, but the fabrication method is not limited to only this one. There can be different ways to fabricate the same injectable dissolvable needle. For example, instead of molding vaccine ink into a vaccine block and then loading the block into the core-shell MP, we can also directly fill the core with vaccine using a pipette. Another example is, instead of using vacuum-thru to mold blocks and needles, we can also centrifuge, etc.

Example 5: Preparation of Further Exemplary Nanoparticle Formulations

The effects of drying processes and solid matrix formulation on mRNA-LNP integrity are poorly understood. While many studies have shown generally that the affinity between polymer materials, excipients, and drug cargo affect diffusion in interstitial fluid, the potential for polymers and other excipients to specifically affect encapsulation efficiency, mRNA-LNP aggregation, or coalescence after re-dissolution has not been well investigated. Studies have primarily focused on functional characterization using in vitro or in vivo assays. Determining an effective framework for characterization of mRNA-LNPs at the nanoscale in the context of re-dissolution is critical to the future development and evaluation of dried, solid matrix mRNA vaccines with high stability for use in low resource settings without cold chain access.

To better understand how drying mRNA-LNPs in a dissolvable polymer matrix affects the integrity and stability of mRNA-LNPs, we performed a series of nanomaterial characterization studies as a function of various system parameters. We report that the stability of mRNA vaccines within a polymer matrix is primarily determined by the inherent characteristics of the LNPs, along with interactions between the polymer and LNPs during drying and redissolution. The affinity between the polymer and mRNA-LNPs induces physiochemical changes in mRNA-LNPs while maintaining mRNA activity. Sufficient polymer is required to prevent LNP aggregation and excessive cholesterol crystallization. A minimum ratio of 160: 1 polymer to mRNA was determined to preserve the integrity of the LNPs and maintain their membrane interaction and luciferase expression in vivo. Future lipid design to bridge the property requirements of high endosomal escape, while simultaneously, exhibiting improved resistance to structural changes observed could lead to future improvements in mRNA-LNP MAP therapies. Analysis of mRNA-LNP structure mRNA-LNPs were prepared via the nanoprecipitation of lipids dissolved in ethanol in an aqueous solution containing mRNA. ILs play a critical role in nanoprecipitation, condensing mRNA into LNPs at low pH during synthesis, protecting mRNA from degradation at neutral pH, and facilitating the release of mRNA once the LNP is internalized by the cell. We therefore sought to investigate the impact of IL to mRNA ratio before, during, and after preparation of vaccine ink. mRNA-LNPs of three different nitrogen-to-phosphate (NP) ratios (1.5, 3.1 and 5.4) were fabricated using firefly luciferase (FLuc) mRNA. mRNA-LNP diameter and the number of mRNA copies per LNP decreased as NP ratio was increased (FIGs. 16A & 16B). In turn, encapsulation efficiency (EE) increased as excess IL allowed for more efficient condensation of mRNA (FIG. 16C). Cryo-TEM of mRNA-LNPs unlabeled and labeled with Au-tagged mRNA showed relatively homogeneous spherically shaped LNPs (FIG. 22).

Characterization of mRNA-LNPs dried in a printable polymer-vaccine ink

Vaccine inks are prepared by mixing mRNA-LNP suspension with dissolved polymers (FIG. 17A). The vaccine inks are then dispensed into microneedle molds, filled into the molds using a vacuum process, and dried to form cohesive devices, such as MAPs (FIG. 17B). Upon application, microneedle tips penetrate the skin and dissolve in the intradermal space, releasing mRNA-LNPs. An in vitro screen previously identified a 50:50 blend of PVP and PVA (PVPPVA) as a formulation that can balance in vitro protein expression with mechanical properties and drying time, with properties that are strongly dependent on the polymer: mRNA ratio in the printed vaccine ink. PVPVA also has direct advantages in vaccine applications due to lower cost, and reduced risk of immune responses associated with impurities in sugars often utilized in lyophilization (Shahad ref).

The effect of PVPVA: mRNA ratio on mRNA-LNPs was studied by mixing PVPVA with mRNA-LNPs at ratios ranging from roughly 10 to 1000. To facilitate characterization, dried films of vaccine ink were prepared in vials for rapid re-dissolution and analysis unless otherwise specified. As the vaccine ink dries, it gradually becomes concentrated, eventually yielding a polymer-composite of mRNA-LNPs embedded within a PVPVA matrix. Since LNP size affects cell uptake, and encapsulation efficiency reflects the successful entrapment of mRNA by the positively charged IL after nanoprecipitation, we assessed these critical parameters before making vaccine inks, within the PVPVA matrix, and after re-dissolution to capture the effect of microneedle matrix drying on mRNA-LNPs.

MAPs with various polymermRNA ratios were sectioned near the tip of a microneedle to allow for examination of LNP size and distribution within the dried composite material (FIG. 17C). Analysis of LNP diameter suggests a polymer:mRNA-ratio-dependent increase in the size of LNPs within the dried matrix, compared to sizing in suspension using DLS (FIGs. 23A-23E). Above 320: 1, PVPVA mitigates LNP aggregation, yielding matrices where the majority of LNPs present are smaller than 500 nm. We used cryo-TEM to evaluate mRNA-LNP morphology in vaccine inks before drying and after re-dissolution (FIG. 17D). Before drying, mRNA-LNPs are consistent in size and morphology across all ratios. mRNA-LNP size quantification using cryo- TEM image analysis shows slight but significant increases compared to mRNA-LNPs alone at high PVPVA ratios before drying and at low PVPVA ratios after drying (FIG. 17E). The PVPVA matrix also locally alters the apparent electron density of LNPs, affecting their contrast in cryo- TEM. mRNA-LNP of different NP ratios were then mixed at different ratios with PVPVA, dried overnight and rehydrated to assess physiochemical properties, EE, and mRNA stability. DLS measurements revealed that the mRNA-LNP size increased significantly after rehydration (FIGs. 23A-23E). Likewise, the size of mRNA-LNPs in PVPVA increased following rehydration and decreased as PVPVA concentration increased (FIG. 18A). Ribogreen (RG) analysis indicates that increasing polymermRNA ratio to >320 improved mRNA recovery (FIGs. 18B-18D). NP ratio 1.5 exhibited a significantly higher free mRNA compared to the other two ratios (FIGs. 18B-18D).

In neat mRNA-LNPs, NP ratio affects particle size (FIG. 17A), encapsulation efficiency (FIG. 17C), and mesostructured, which may have downstream effects on the mRNA-LNPs after re-dissolution from vaccine ink. While all NP ratios showed a high EE (>60%) at polymer: mRNA ratios greater than 320, total mRNA recovery was lower in NP 5.4 compared to the two other ratios (FIGs. 18B-18D). However, NP 5.4 exhibited higher EE than other ratios (FIG. 18E), indicating the role of IL in maintaining high EE. mRNA integrity was also assessed using capillary gel electrophoresis (FIG. 18F) The poor LNP stability at low polymer: mRNA ratios results in more unencapsulated mRNA, and the mRNA present is more susceptible to degradation. Collectively, mRNA integrity is preserved at polymermRNA ratios greater than 160. During drying the colloidal solution becomes concentrated, decreasing the separation between mRNA-LNPs, and inducing changes in lipid fluidity as water leaves the system. Within 12 hours, almost all water has been removed the ink (FIG. 24A) and encapsulation efficiency drops starkly for formulations without PVPVA, despite particle size remaining relatively consistent (FIGs. 24B-24D). mRNA-LNPs in PVPVA are also sensitive to the geometry of the mold used for drying, with increased mold surface area increasing efficiency of redissolution (FIG. 25A) but decreasing LNP encapsulation efficiency (FIG. 25B)

The relative distribution of LNPs of empty LNPs was correlated to NP ratio, while mRNA copies per particle was inversely correlated to NP ratio (FIG. 16B). As a result, in formulations with higher NP ratios coalescence may be more likely to occur with non-mRNA loaded LNPs. Conversely, mRNA-loaded LNPs were more likely to fuse together at lower NP ratios. This in part results in the expulsion of mRNA during the coalescence process. As such, coalescence at higher NP ratios is less likely to reduce the number of mRNA-loaded carriers.

Cryo-TEM shows the variety mRNA-LNPs morphologies present after MAP matrix redissolution (FIG. 19A-19F). In some instances, small spheres fusing with the LNP (FIG. 19A), or blebs (FIG. 19E), or extended protrusions are observed (FIG. 19C).

In terms of total dye delivered, NP2 delivered less fluorescent dye to the membrane than NP4 and NP8 for both neat and rehydrated LNPs (FIG. 20A). In vivo imaging study of luciferase following administration of mRNA-LNPs loaded in MAPs at varying polymermRNA ratios indicates a significant leap in flux from increasing the polymer: mRNA from 32 to 160+ (FIG. 5D & 5E). Beyond this ratio, performance was similar, in general agreement with colloidal characterization in FIGs. 17A-17E, 18A-18F, and 19A-19F.

MAPs require a polymeric excipient such as PVPVA to give individual microneedles enough strength to penetrate the skin. However, several other excipients for mRNA-LNPs based on blends of sugars such as sucrose, trehalose, and maltose have been studied for lyophilization (references). To compare the effectiveness of PVPVA a promising sucrose/maltose blend and to PBS buffer, we prepared films of mRNA-LNPs from these three different formulations and analyzed their ability to preserve mRNA-LNP integrity before and after drying (FIGs. 27A-27D), and after one month of storage at three different temperatures (FIGs. 27E-27G). While LNP size was preserved across all formulations, LNPs in PVPVA were redissolved more efficiently and maintained their encapsulation efficiency and in vivo activity throughout the process. For PVPVA, lyophilization was comparable to vacuum drying, indicating that the microneedle mold fabrication process, which uses vacuum drying to remove water from the vaccine ink, does not inherently inhibit mRNA-LNP activity. After drying, both PVPVA and sucrose/maltose were stable for one month of storage.

Optimizing Microneedle Patch Fabrication

Conventionally, MAPs often integrate several fabrication steps with multiple drying cycles to create complex, multilayered microneedle structures and to reduce drug waste (references). However, because of the apparent sensitivity of lipids within mRNA-LNPs to dehydration, converting MAP fabrication to a process with a single drying step (FIG. 21A) yields several benefits. While LNP size and encapsulated mRNA loading is not significantly affected (FIGs. 21B & 21C), RNA integrity decreases dramatically after the second drying cycle (FIG. 21D), perhaps due to the structural changes described above. The structure of the MAPs is reinforced by the backing, but using a concentrated ink can overcome the tendency to produce partially filled needles (FIG. 21E), resulting in mechanically stable MAPs fabricated without a backing step (FIG. 21).

Despite holding a similar amount of mRNA in the microneedles, mRNA MAPs fabricated with a one drying cycle process induce significantly greater Flue expression in BALB/c mice than those made with two drying cycles and a backing (FIG. 21). We then compared the immunogenicity of one cycle MAPs loaded with mRNA expressing the SARS-CoV-2 S protein receptor binding domain to increasing doses of the same mRNA administered intramuscularly (FIG. 21G) or intradermally (FIG. 29). Finally, we developed microneedle patches loaded with more mRNA by using a larger volume of vaccine ink on a larger microneedle mold (FIG. 21H), and vaccinated Wistar rats with two patches containing two different doses (FIG. 211 & 21J). These produce SARS-CoV-2 pseudo virus neutralizing titers comparable to IM injections of similar or greater doses.

INCORPORATION BY REFERENCE

All US and PCT patent application publications and US patents mentioned herein are hereby incorporated by reference in their entirety as if each individual patent application publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control. EQUIVALENTS

While specific embodiments of the subject invention have been discussed, the above specification is illustrative and not restrictive. Many variations of the invention will become apparent to those skilled in the art upon review of this specification and the claims below. The full scope of the invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations.

Claims

CLAIMS We claim:

1. An injectable needle, comprising a needle body, a needle tip, a compartment, a first composition, and a second composition; wherein: the needle body and the needle tip are connected to one another; the first composition and the compartment are encapsulated by the needle tip and the needle body; the compartment comprises a compartment base and a compartment cap; and the second composition is encapsulated in the compartment.

2. The injectable needle of claim 1, wherein the length of the injectable needle is 1 - 10 mm.

3. The injectable needle of claim 1, wherein the length of the injectable needle is about 1 mm, about 2 mm, about 3 mm, about 4 mm, about 5 mm, or about 6 mm.

4. The injectable needle of claim 1, wherein the length of the injectable needle is about 4 mm, about 5 mm, or about 6 mm.

5. The injectable needle of any one of claims 1-4, wherein the width of the injectable needle is 100 - 1,500 pm.

6. The injectable needle of any one of claims 1-4, wherein the width of the injectable needle is about 200 pm, about 300 pm, about 400 pm, about 500 pm, about 600 pm, about 700 pm, about 800 pm, about 900 pm, about 1,000 pm, about 1,100 pm, about 1,200 pm, about 1,300 pm, about 1,400 pm, or about 1,500 pm.

7. The injectable needle of any one of claims 1-4, wherein the width of the injectable needle is about 450 pm.

8. The injectable needle of any one of claims 1-7, wherein the height of the injectable needle is 100 - 1,000 gm.

9. The injectable needle of any one of claims 1-7, wherein the height of the injectable needle is about 200 gm, about 300 gm, about 400 gm, about 500 gm, or about 600 gm.

10. The injectable needle of any one of claims 1-7, wherein the height of the injectable needle is about 450 gm.

11. The injectable needle of any one of claims 1-10, wherein the injectable needle is a 16 gauge, 18 gauge, 19 gauge, 20 gauge, 21 gauge, 22 gauge, 23 gauge needle, 28 gauge, or 30 gauge needle.

12. The injectable needle of any one of claims 1-10, wherein the injectable needle is a 21 gauge, 22 gauge, or 23 gauge needle.

13. The injectable needle of any one of claims 1-12, wherein the needle body comprises a polymer or a copolymer.

14. The injectable needle of any one of claims 1-12, wherein the needle body comprises a blend of polymers (e.g., a blend of 2, 3, 4, or 5 polymers, preferably a blend of 2 polymers).

15. The injectable needle of any one of claims 1-12, wherein the needle body comprises a blend of polyvinylpyrrolidone (PVP) and polyvinylalcohol (PVA), or a blend of PVP, PVA, and sucrose; preferably a blend of PVP and PVA.

16. The injectable needle of claim 15, wherein the PVP is PVP10 (i.e., PVP with an average molecular weight of 10 kDa) or PVP60 (i.e., PVP with an average molecular weight of 60 kDa).

17. The injectable needle of claim 15 or 16, wherein the ratio of PVP:PVA in the polymer blend is 1 :1, 2: 1, or 5:1.

18. The injectable needle of any one of claims 1-17, wherein the needle body comprises a copolymer comprising PVP60 and PVA at a ratio of 1 : 1.

19. The injectable needle of claim 13, wherein the needle body comprises a random copolymer.

20. The injectable needle of claim 13, wherein the needle body comprises a block copolymer.

21. The injectable needle of any one of claims 1-20, wherein the needle tip comprises a polymer or a copolymer.

22. The injectable needle of any one of claims 1-20, wherein the needle tip comprises a blend of polymers (e.g., a blend of 2, 3, 4, or 5 polymers, preferably a blend of 2 polymers).

23. The injectable needle of any one of claims 1-20, wherein the needle tip comprises a blend of PVP and PVA or a blend of PVP, PVA, and sucrose, preferably a blend of PVP and PVA.

24. The injectable needle of claim 23, wherein the PVP is PVP10 (i.e., PVP with an average molecular weight of 10 kDa) or PVP60 (i.e., PVP with an average molecular weight of 60 kDa).

25. The injectable needle of claim 23, wherein the PVP is PVP10.

26. The injectable needle of claim 23, wherein the PVP is PVP60.

27. The injectable needle of any one of claims 24-26, wherein the ratio of PVP:PVA in the polymer blend is 1: 1, 2:1, or 5: 1.

28. The injectable needle of any one of claims 24-26, wherein the w/w ratio of PVP:PVA in the polymer blend is 1: 1, 2:1, or 5: 1.

29. The injectable needle of any one of claims 1-23, wherein the needle tip comprises a blend of PVP60 and PVA at a w/w ratio of 5: 1.

30. The injectable needle of any one of claims 1-23, wherein the needle tip comprises a blend of PVP10 and PVA at a w/w ratio of 1 : 1.

31. The injectable needle of any one of claims 1-30, wherein the needle tip comprises a block copolymer.

32. The injectable needle of any one of claims 1-30, wherein the needle tip comprises a random copolymer.

33. The injectable needle of any one of claims 1-32, wherein the first composition is encapsulated in the needle tip.

34. The injectable needle of any one of claims 1-33, wherein the compartment base comprises a polymer.

35. The injectable needle of any one of claims 1-33 wherein the compartment base comprises a copolymer.

36. The injectable needle of any one of claims 1-33, wherein the compartment base comprises poly lactic-co-gly colic acid (PLGA).

37. The injectable needle of any one of claims 1-33, wherein the compartment base comprises PLGA 858 (i.e., a copolymer of lactide: glycolide at a w/w ratio of 85: 15).

38. The injectable needle of any one of claims 1-37, wherein the compartment base comprises a random copolymer.

39. The injectable needle of any one of claims 1-37, wherein the compartment base comprises a block copolymer.

40. The injectable needle of any one of claims 1-29, wherein the compartment cap comprises PLGA 502 (i.e., a copolymer of lactide: glycolide at a w/w ratio of 1 : 1).

41. The injectable needle of any one of claims 1-40, wherein the compartment cap comprises a random copolymer.

42. The injectable needle of any one of claims 1-40, wherein the compartment cap comprises a block copolymer.

43. The injectable needle of any one of claims 1-42, wherein the first composition or the second composition comprises a nanoparticle (e.g., a polymeric nanoparticle of a gold nanoparticle or a lipid nanoparticle).

44. The injectable needle of any one of claims 1-42, wherein the first composition comprises a nanoparticle (e.g., a polymeric nanoparticle of a gold nanoparticle or a lipid nanoparticle).

45. The injectable needle of any one of claims 1-42, wherein the second composition comprises a nanoparticle (e.g., a polymeric nanoparticle of a gold nanoparticle or a lipid nanoparticle).

46. The injectable needle of any one of claims 1-42, wherein the first composition or the second composition comprises a lipid nanoparticle comprising an ionizable lipid, a PEG lipid, a phospholipid, or a combination of the foregoing.

47. The injectable needle of any one of claims 1-42, wherein the first composition comprises a lipid nanoparticle comprising an ionizable lipid, a PEG lipid, a phospholipid, or a combination of the foregoing.

48. The injectable needle of any one of claims 1-42, wherein the second composition comprises a lipid nanoparticle comprising an ionizable lipid, a PEG lipid, a phospholipid, or a combination of the foregoing.

49. The injectable needle of any one of claims 1-48, wherein the first composition or the second composition comprises: i) a polymer or a copolymer and a lipid nanoparticle, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is at least 100: 1; or ii) a polymer or a copolymer and a bioactive agent, wherein the w/w ratio of the polymer or the copolymer to the bioactive agent is at least 1000: 1.

50. The injectable needle of claim 49, wherein the first composition or the second composition comprises a polymer or a copolymer and a lipid nanoparticle, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is at least 100: 1.

51. The injectable needle of claim 49, wherein the first composition comprises a polymer or a copolymer and a lipid nanoparticle, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is at least 100: 1.

52. The injectable needle of claim 49, wherein the second composition comprises a polymer or a copolymer and a lipid nanoparticle, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is at least 100:1.

53. The injectable needle of any one of claims 49-52, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is about 100: 1, 150: 1, 200:1, 250: 1, 300: 1, 350:1, 400: 1, 450: 1, or 500: 1.

54. The injectable needle of claim 53, wherein the w/w ratio of the polymer or the copolymer to the lipid nanoparticle is about 333: 1.

55. The injectable needle of any one of claims 43-54, wherein the lipid nanoparticle comprises an ionizable lipid, cholesterol, a polyethyleneglcyol lipid, a phospholipid, or a combination thereof.

56. The injectable needle of any one of claims 43-55, wherein the lipid nanoparticle comprises 1-octylnonyl 8-[(2-hydroxyethyl)[8-(nonyloxy)-8-oxooctyl]amino]octanoate (i.e., lipid 5, CAS Ref. No.: 2089251-33-0).

57. The injectable needle of any one of claims 43-55, wherein the lipid nanoparticle consists essentially of 1-octylnonyl 8-[(2-hydroxyethyl)[8-(nonyloxy)-8-oxooctyl]amino]octanoate (i.e., lipid 5).

58. The injectable needle of any one of claims 43-55, wherein the lipid nanoparticle comprises 3, 6-bis(4-(bis(2-hydroxydodecyl)amino)butyl)piperazine-2, 5-dione (i.e., cKK-E12).

59. The injectable needle of any one of claims 43-55, wherein the lipid nanoparticle consists essentially of 3, 6-bis(4-(bis(2-hydroxydodecyl)amino)butyl)piperazine-2, 5-dione (i.e., cKK- E12).

60. The injectable needle of any one of claims 43-59, wherein the diameter of the lipid nanoparticle is about 50 nm, about 75 nm, about 100 nm, about 125 nm, about 150 nm, about 175 nm, about 200 nm, about 225 nm, about 250, about 275 nm, or about 300 nm.

61. The injectable needle of any one of claims 43-59, wherein the diameter of the lipid nanoparticle is about 125 nm.

62. The injectable needle of any one of claims 43-59, wherein the diameter of the lipid nanoparticle is about 225 nm.

63. The injectable needle of any one of claims 43-62, further comprising a bioactive agent.

64. The injectable needle of claim 63, wherein the w/w ratio of the polymer or the copolymer to the bioactive agent is at least 1000: 1.

65. The injectable needle of claim 63, wherein the w/w ratio of the polymer or the copolymer to the bioactive agent is about 600:1, about 700:1, about 800:1, 900: 1, or about 1000: 1.

66. The injectable needle of claim 63, wherein the w/w ratio of the polymer or the copolymer to the bioactive agent is about 1,250:1, about 1500: 1, about 1750: 1, about 2000:1, about 2,250: 1, about 2,500:1, about 2,750: 1, or about 3000:1.

67. The injectable needle of claim 63, wherein the w/w ratio of the polymer or the copolymer to the bioactive agent is about 4,000:1, about 5,000:1, about 6,000:1, about 7000: 1, about 8,000: 1, about 9,000: 1, or about 10,000: 1.

68. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is about 20:1, about 40: 1, about 60: 1, about 80:1, about 100:1, 120: 1, 140:1, 160: 1, 180: 1, 200:1, 220: 1, 240: 1, 260: 1, 280:1, or about 300: 1, about 320: 1, about 340: 1, about 360: 1, about 380: 1, or about 400:1.

69. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is about 50:1.

70. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is about 160: 1.

71. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is about 320: 1.

72. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is greater than about 100: 1.

73. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is greater than about 150: 1.

74. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is greater than about 200: 1.

75. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is greater than about 250: 1.

76. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is greater than about 300: 1.

77. The injectable needle of any one of claims 63-67, wherein the w/w ratio of copolymer to bioactive agent is greater than about 320: 1.

78. The injectable needle of any one of claims 1-77, wherein the first composition or the second composition further comprises a polymer.

79. The injectable needle of any one of claims 1-77, wherein the first composition further comprises a polymer.

80. The injectable needle of any one of claims 1-77, wherein the second composition further comprises a polymer.

81. The injectable needle of claim 80, wherein the polymer is PVP.

82. The injectable needle of claim 80, wherein the PVP is PVP10.

83. The injectable needle of claim 80, wherein the PVP is PVP60.

84. The injectable needle of claim 80, wherein the polymer is polyvinylalcohol.

85. The injectable needle of any one of claims 1-77, wherein the first composition or the second composition further comprises a copolymer.

86. The injectable needle of any one of claims 1-77, wherein the first composition further comprises a copolymer.

87. The injectable needle of any one of claims 1-77, wherein the second composition further comprises a copolymer.

88. The injectable needle of claim 87, wherein the copolymer comprises a plurality of repeat units of vinylalcohol; a plurality of repeat units of vinylpyrrolidinone; and the copolymer is a block copolymer or a random copolymer.

89. The injectable needle of claim 87, wherein the copolymer consists essentially of a plurality of repeat units of vinylalcohol; and a plurality of repeat units of vinylpyrrolidinone; and the copolymer is a block copolymer or a random copolymer.

90. The injectable needle of claim 87 or 88, wherein the mass ratio of polyvinylalcohol to polyvinylpyrrolidone is about 1: 1, about 2: 1, about 3:1, about 4: 1, about 5:1, or about 6: 1.

91. The injectable needle of claim 87 or 88, wherein the mass ratio of polyvinylalcohol to polyvinylpyrrolidone is about 1: 1, about 2: 1, or about 3: 1.

92. The injectable needle of claim 87 or 88, wherein the mass ratio of polyvinylalcohol to polyvinylpyrrolidone is about 1: 1.

93. The injectable needle of claim 87 or 88, wherein the mass ratio of polyvinylalcohol to polyvinylpyrrolidone is about 2: 1.

94. The injectable needle of claim 87, wherein the copolymer comprises a plurality of repeat units of vinylalcohol; a plurality of repeat units of sucrose; and the copolymer is a block copolymer or a random copolymer.

95. The injectable needle of claim 87, wherein the copolymer consists essentially of a plurality of repeat units of vinylalcohol; a plurality of repeat units of sucrose; and the copolymer is a block copolymer or a random copolymer.

96. The injectable needle of claim 94 or 95, wherein the mass ratio of polyvinylalcohol to sucrose is about 1 :1, about 2:1, about 3: 1, about 4: 1, about 5:1, or about 6:1.

97. The injectable needle of claim 94 or 95, wherein the mass ratio of polyvinylalcohol to sucrose is about 1 : 1 or about 2:1.

98. The injectable needle of claim 87, wherein the copolymer comprises a plurality of repeat units of vinylalcohol, a plurality of repeat units of polyvinylpyrrolidone, a plurality of repeat units of sucrose; and the copolymer is a block copolymer or a random copolymer.

99. The injectable needle of claim 87, wherein the copolymer consists essentially of a plurality of repeat units derived from vinylalcohol, a plurality of repeat units derived from polyvinylpyrrolidone, a plurality of repeat units derived from sucrose; and the copolymer is a block copolymer or a random copolymer.

100. The injectable needle of claim 98 or 99, wherein the mass ratio of polyvinylalcohol to polyvinylpyrrolidone to sucrose is about 1: 1: 1, about 1 :1 :2, or about 1 :1 :3.

101. The injectable needle of claim 98 or 99, wherein the mass ratio of polyvinylalcohol to polyvinylpyrrolidone to sucrose is about 1: 1:2.

102. The injectable needle of any one of claims 85-101, wherein the copolymer is a block copolymer.

103. The injectable needle of any one of claims 85-101, wherein the copolymer is a random copolymer.

104. The injectable needle of any one of claims 38-85, wherein the polymer comprises 50 - 500 repeat units.

105. The injectable needle of any one of claims 38-85, wherein the polymer comprises 50 - 250 repeat units.

106. The injectable needle of any one of claims 38-85, wherein the polymer comprises 75 - 125 repeat units.

107. The injectable needle of any one of claims 38-85, wherein the polymer comprises about 70, about 80, about 90, about 100, about 110, or about 120 repeat units.

108. The injectable needle of any one of claims 38-85, wherein the polymer comprises about 90 repeat units.

109. The injectable needle of any one of claims 87-103, wherein the copolymer comprises 250

- 1,500 repeat units.

110. The injectable needle of any one of claims 87-103, wherein the copolymer comprises 500

- 1250 repeat units.

111. The injectable needle of any one of claims 87-103, wherein the copolymer comprises 500

- 1000 repeat units.

112. The injectable needle of any one of claims 87-103, wherein the copolymer comprises 600

- 800 repeat units.

113. The injectable needle of any one of claims 87-103, wherein the copolymer comprises about 600, about 650, about 700, about 750, about 800, about 850, or about 900 repeat units.

114. The injectable needle of any one of claims 87-103, wherein the copolymer comprises about 700 repeat units.

115. The injectable needle of any one of claims 1-114, wherein the first composition or the second composition further comprises a bioactive agent.

116. The injectable needle of any one of claims 1-114, wherein the first composition further comprises a bioactive agent.

117. The injectable needle of any one of claims 1-114, wherein the second composition further comprises a bioactive agent.

118. The injectable needle of any one of claims 49-117, wherein the concentration of the bioactive agent is about 50 pg/mL, about 100 pg/mL, about 150 pg/mL, about 200 pg/mL, about 250 pg/mL, about 300 pg/mL, about 350 pg/mL, about 400 pg/mL, about 450 pg/mL, about 500 pg/mL, about 550 pg/mL, about 600 pg/mL, about 650 pg/mL about 700 pg/mL, about 750 pg/mL, about 800 pg/mL, about 850 pg/mL, about 900 pg/mL, about 950 pg/mL or about 1,000 pg/mL.

119. The injectable needle of any one of claims 49-117, wherein the concentration of the bioactive agent is about 650 pg/mL, about 700 pg/mL, or about 750 pg/mL.

120. The injectable needle of any one of claims 49-117, wherein the concentration of the bioactive agent is about 700 pg/mL.

121. The injectable needle of any one of claims 49-117, wherein the weight of the bioactive agent is about 2 pg, about 4 pg, about 6 pg, about 8 pg, about 10 pg, about 12 pg, or about 14 pg, about 16 pg.

122. The injectable needle of any one of claims 49-117, wherein the weight of the bioactive agent is about 6 pg.

123. The injectable needle of any one of claims 49-117, wherein the weight of the bioactive agent is about 7 pg.

124. The injectable needle of any one of claims 49-123, wherein the w/w ratio of copolymer to bioactive agent is about 20:1, about 40: 1, about 60: 1, about 80:1, about 100:1, 120: 1, 140:1, 160: 1, 180: 1, 200:1, 220: 1, 240: 1, 260: 1, 280:1, or about 300: 1, about 320: 1, about 340: 1, about 360: 1, about 380: 1, or about 400:1.

125. The injectable needle of any one of claims 49-123, wherein the w/w ratio of copolymer to bioactive agent is about 50:1.

126. The injectable needle of any one of claims 49-123, wherein the w/w ratio of copolymer to bioactive agent is about 160: 1.

127. The injectable needle of any one of claims 49-123, wherein the w/w ratio of copolymer to bioactive agent is about 320: 1.

128. The injectable needle of any one of claims 49-123, wherein the w/w ratio of copolymer to bioactive agent is greater than about 100: 1.

129. The injectable needle of any one of claims 49-123, wherein the w/w ratio of copolymer to bioactive agent is greater than about 150: 1.

130. The injectable needle of any one of claims 49-123, wherein the w/w ratio of copolymer to bioactive agent is greater than about 200: 1.

131. The injectable needle of any one of claims 49-123, wherein the w/w ratio of copolymer to bioactive agent is greater than about 250: 1.

132. The injectable needle of any one of claims 49-123, wherein the w/w ratio of copolymer to bioactive agent is greater than about 300: 1.

133. The injectable needle of any one of claims 43-132, wherein the w/w ratio of copolymer to bioactive agent is greater than about 320: 1.

134. The injectable needle of any one of claims 43-132, wherein the nitrogen to phosphate ratio of the lipid nanoparticle is about 1.5.

135. The injectable needle of any one of claims 43-132, wherein the nitrogen to phosphate ratio of the lipid nanoparticle is about 3.1.

136. The injectable needle of any one of claims 43-132, wherein the nitrogen to phosphate ratio of the lipid nanoparticle is about 5.4

137. The injectable needle of any one of claims 43-136, wherein the encapsulation efficiency of the lipid nanoparticle is greater than about 40%, about 50%, about 60%, about 70%, about 80%, or about 90%.

138. The injectable needle of any one of claims 49-136, wherein the encapsulation efficiency of the lipid nanoparticle is greater than about 60%.

139. The injectable needle of any one of claims 49-138, wherein the bioactive agent has improved stability (e.g., improved thermostability) as compared to the bioactive agent alone or the bioactive agent in a composition not recited in any one of claims 49-138.

140. The injectable needle of any one of claims 49-139, wherein each bioactive agent is independently selected from the group consisting of a protein, an antibody, a nucleic acid, a small molecule, and a vaccine.

141. The injectable needle of any one of claims 49-140, wherein at least one bioactive agent is a mRNA, siRNA, RNA, or DNA.

142. The injectable needle of any one of claims 49-140, wherein the at least one bioactive agent is an antibody.

143. The injectable needle of any one of claims 49-140, wherein the at least one bioactive agent is a small molecule (e.g., a drug).

144. The injectable needle of any one of claims 49-140, wherein the at least one bioactive agent is a protein.

145. The injectable needle of any one of claims 49-140, wherein the at least one bioactive agent is a vaccine.

146. The injectable needle of any one of claims 49-140, wherein the at least one bioactive agent is a nucleic acid.

147. The injectable needle of any one of claims 49-140, wherein the at least one bioactive agent is an RNA (e.g., an mRNA).

148. The injectable needle of any one of claims 1-147, wherein the first composition or the second composition further comprises a pharmaceutically acceptable carrier.

149. The injectable needle of any one of claims 1-148, wherein the first composition or the second composition further comprises an adjuvant.

150. The injectable needle of any one of claims 1-149, wherein the injectable needle is dissolvable.

151. The injectable needle of any one of claims 1-150, wherein the injectable needle has been formed using a single drying step.

152. The injectable needle of any one of claims 1-12, wherein: the needle body comprises a copolymer comprising PVP60 and PVA at a w/w ratio of 1 : 1 ; the needle tip comprises a blend of PVP10 and PVA at a w/w ratio of 1 : 1; the compartment base comprises PLGA 858 (i.e., a copolymer of lactide: glycolide at a ratio of w/w 85: 15); the compartment cap comprises PLGA 502 (i.e., a copolymer of lactide: glycolide at a ratio of 1: 1); the first composition comprises:

PVP 10 and PVA at a w/w ratio of 1 : 1 ; and a bioactive agent, further wherein: the weight of the bioactive agent is about 7 pg; and the w/w ratio of copolymer to bioactive agent is about 320: 1 ; the second composition comprises:

PVP 10 and PVA at a w/w ratio of 1 :2; a bioactive agent, further wherein: the weight of the bioactive agent is about 6 pg; and the w/w ratio of copolymer to bioactive agent is about 50: 1.

-SO-

153. The injectable needle of claim 152, wherein the bioactive agent is an RNA (e.g., an mRNA).

154. A method of delivering a therapy to a subject in need thereof, comprising contacting the subject with the injectable needle of any one of claims 1-153.

155. A method of delivering a therapy to a subject in need thereof, consisting essentially of contacting the subject with the injectable needle of any one of claims 1-153.

156. The method of claims 154 or 155, wherein the therapy is a vaccine.

157. The method of claims 154 or 155, wherein the therapy is an mRNA vaccine.

158. The method of claim any one of claims 154-157, wherein the therapy is an anticancer therapy, an antibacterial therapy, or an anticancer therapy.

159. The method of any one of claims 154-158, wherein the therapy is delivered in the form of a first dose and a second dose.

160. The method of claim 159, wherein the first dose and the second dose are delivered 1 day to six months apart.

161. The method of claim 159, wherein the first dose and the second dose are delivered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 days apart.

162. The method of any one of claims 154-161, wherein the therapy is delivered in the form of a plurality of doses (e.g., 2, 3, 4, or more doses).