Method of intranasal delivery of lipid nanoparticles

Abstract

The present disclosure provides devices and methods for intranasal delivery of a formulation comprising a delivery vehicle. Devices and methods of the present disclosure may provide convenient and effective ways to accomplish targeted intranasal delivery of various delivery vehicles.

Description

Mintz Ref.: 062706-519001 WOMETHOD OF INTRANASAL DELIVERY OF LIPID NANOPARTICLESCROSS REFERENCE

[0001] This PCT application claims the benefit to U.S. Provisional Application No. 63 / 725,789, filed November 27, 2024, U.S. Provisional Application No. 63 / 866,511, filed August 19, 2025, the contents of which are incorporated herein in their entirety by reference.FIELD

[0002] The present disclosure relates to methods of delivering delivery vehicles, more specifically, methods of delivering delivery vehicles intranasally.BACKGROUND

[0003] Delivery vehicles can be used to deliver drugs and other molecules to a subject. The use of a delivery vehicle can improve stability of the molecules, reduce toxicity, etc. For example, lipid nanoparticles can improve the stability of biologic drugs and can direct targeting to specific sites, improving their efficacy and reducing the quantity of the biologic needed for the desired therapeutic effects. There remains a need for improved methods of delivering delivery vehicles to a subject.SUMMARY

[0004] In one aspect, provided herein is a method of delivering a delivery vehicle to a subject in need thereof, the method comprising: delivering a formulation comprising the delivery vehicle to a targeted region of a nasal cavity of the subject, wherein the formulation is delivered as a liquid jet.

[0005] In some embodiments, the liquid jet is a laminar jet.

[0006] In some embodiments, the delivery vehicle comprises a macromolecule delivery vehicle.

[0007] In some embodiments, the delivery vehicle comprises an exosome.

[0008] In some embodiments, the delivery vehicle comprises a nucleic acid delivery vehicle.

[0009] In some embodiments, the nucleic acid delivery vehicle comprises a viral vector.

[0010] In some embodiments, the viral vector comprises an adeno-associated vector (AVV).

[0011] In some embodiments, the delivery vehicle comprises a lipid nanoparticle.

[0012] In some embodiments, the lipid nanoparticle comprises a nucleic acid molecule.

[0013] In some embodiments, the lipid nanoparticle comprises a vaccine.

[0014] In some embodiments, administering of the formulation results in an increased antigenicity in the subject compared to where the formulation is delivered to the subject as a spray at about the same dose.Mintz Ref.: 062706-519001 WO

[0015] In some embodiments, the delivery vehicle comprises a cargo.

[0016] In some embodiments, the cargo comprises a nucleic acid molecule, a small molecule, a protein, a peptide, or any combination thereof.

[0017] In some embodiments, the formulation is delivered by a device, the device comprising: (a) a housing comprising an insertable portion comprising a distal end, and a proximal end; and (b) a subject-engaging portion which engages a columella region of the subject to seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject, wherein the device is configured to deliver the formulation as a laminar jet.

[0018] In one aspect, provided herein is a device for intranasal delivery of a delivery vehicle to a targeted region of a nasal cavity of a subject, the device comprising: (a) a housing comprising an insertable portion comprising a distal end, and a proximal end; and (b) a subject-engaging portion which engages a columella region of the subject to seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject; wherein the device is configured to deliver a formulation comprising the delivery vehicle to the targeted region of the subject.

[0019] In some embodiments, the device dispenses the formulation as a laminar jet.

[0020] In some embodiments, the device comprises a compliant dispensing tip comprising a compliant and flexible soft nib.

[0021] In some embodiments, the device is configured to deliver the formulation at a velocity of between about 1 m / s and about 30 m / s or between about 1 m / s and about 5 m / s.

[0022] In some embodiments, application of pressure by the subject-engaging portion to the columella region of the subject enables and / or causes delivery of the formulation to the subject from the insertable portion.

[0023] In some embodiments, the insertable portion comprises a dispensing element for delivery of the formulation to the targeted region of the subject.

[0024] In some embodiments, the device is configured to deliver at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% of the formulation to the targeted region of the nasal cavity of the subject.

[0025] In some embodiments, the ejection zone is: (a) 0mm to 30mm superior to a horizontal line that intersects the anterior aspect of the internal nasal valve, and (b) 0mm to 20mm anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule.

[0026] Other features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and theMintz Ref.: 062706-519001 WO specific examples while indicating preferred embodiments of the disclosure are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description.INCORPORATION BY REFERENCE

[0027] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The novel features of the present disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description and accompanying drawings, that set forth illustrative embodiments in which the principles of the present disclosure are utilized.

[0029] FIG. 1A depicts a bottom view of an exemplary embodiment of a subject’s nose.

[0030] FIG. IB depicts a side view of an exemplary embodiment of a subject’s nose.

[0031] FIG. 1C depicts a side view of a representative subject’s nasal channel.

[0032] FIG. ID depicts a side view of an exemplary embodiment of a representative subject’s nasal channels from the vestibule to the olfactory cleft based on a posteriorly oriented plane.

[0033] FIG. IE depicts a side view of an exemplary embodiment of a representative subject’s nasal channels from the vestibule to the olfactory cleft based on an anteriorly oriented plane.

[0034] FIG. IF depicts a side view of an exemplary embodiment of a representative target ejection zone, according to some embodiments.

[0035] FIG. 1G depicts a front, base and perspective view of an exemplary embodiment of a subject’s nose and the columella region.

[0036] FIG. 1H depicts a side view of an exemplary embodiment of a representative subject’s nasal cavity, including internal anatomical features.

[0037] FIG. II depicts a side view of an exemplary embodiment of a representative subject’s sinus, according to some embodiments.

[0038] FIG. 1J depicts a side view of a subject’s nasal cavity.

[0039] FIG. 2A depicts an exemplary embodiment of an Exemplary Device in a second configuration, according to some embodiments.Mintz Ref.: 062706-519001 WO

[0040] FIG. 2B depicts an exemplary embodiment top view of an Exemplary Device in the second configuration, according to some embodiments.

[0041] FIG. 2C depicts an exemplary embodiment trigger button side view of an Exemplary Device in the second configuration, according to some embodiments.

[0042] FIG. 2D depicts an exemplary embodiment chassis side view of an Exemplary Device in the second configuration, according to some embodiments.

[0043] FIG. 2E depicts an exemplary embodiment bottom view of an Exemplary Device in the second configuration, according to some embodiments.

[0044] FIG. 2F depicts an exemplary embodiment perspective view of an Exemplary Device in the first configuration, according to some embodiments.

[0045] FIG. 2G depicts an exemplary embodiment front view of another Exemplary Device in the first configuration, according to some embodiments.

[0046] FIG. 2H depicts an exemplary embodiment front view of an Exemplary Device in the second configuration, according to some embodiments.

[0047] FIG. 21 depicts an exemplary embodiment front view of another Exemplary Device in the first configuration, according to some embodiments.

[0048] FIG. 2J depicts an exemplary embodiment front view of an Exemplary Device in the second configuration, according to some embodiments.

[0049] FIG. 3A depicts an exemplary embodiment of an Exemplary Device in the second configuration with one dispensing element revealed and a side view of an exemplary embodiment of a subject’s nose, according to some embodiments.

[0050] FIG. 3B depicts an exemplary embodiment of an Exemplary Device in the second configuration with one dispensing element revealing along the passageway and a side view of an exemplary embodiment of a representative subject’s nasal channels from the vestibule to the olfactory cleft based on an anteriorly oriented plane, according to some embodiments.

[0051] FIG. 4A depicts an exemplary embodiment of a cartesian reference plane of (left to right) a front, a top, and a side view of an Exemplary Device, according to some embodiments.

[0052] FIG. 4B depicts an exemplary embodiment of the subject plane, according to some embodiments.

[0053] FIG. 4C depicts an exemplary embodiment of (left to right) a front Coronal Plane, a top Transverse Plane, and a back Sagittal Plane view, according to some embodiments.Mintz Ref.: 062706-519001 WO

[0054] FIG. 4D depicts an exemplary embodiment of an Exemplary Device Sagittal Angle positioning in the subject, according to some embodiments.

[0055] FIG. 4E depicts an exemplary embodiment of an Exemplary Device Coronal-Medial Angle positioning in the subject, along a front view of the Coronal Plane, and a top view along a transverse plane, according to some embodiments.

[0056] FIG. 4F depicts an exemplary embodiment of an Exemplary Device Depth positioning along a front view of the Coronal Plane, and along a side view of the Sagittal Plane in the subject, according to some embodiments.

[0057] FIG. 4G depicts an exemplary embodiment of an Exemplary Device along a side view of the sagittal plane of delivery to the target region in the subject, according to some embodiments.

[0058] FIG. 4H depicts an exemplary embodiment of an Exemplary Device along a side view of the sagittal plane aiming from the respiratory region in the subject, according to some embodiments.

[0059] FIG. 41 depicts a side view of a target ejection zone, according to some embodiments.

[0060] FIG. 4J depicts a side view of a target ejection zone with respect to other nasal cavity anatomy, according to some embodiments.

[0061] FIGS. 5A-C depict images of a transparent model nasal cavity with a colored solution demonstrating delivery to an olfactory region of the model.

[0062] FIG. 6A depicts a mouse in an olfactory delivery study with a catheter inserted into the nostril to deliver a formulation to the olfactory region.

[0063] FIG. 6B depicts a diagram of the targeted olfactory delivery shown in FIG. 6A.

[0064] FIG. 7A depicts a mouse in an intranasal delivery study with a pipet tip inserted into the nostril to deliver a formulation to the nasal cavity.

[0065] FIG. 7B depicts a diagram of the non-targeted intranasal delivery shown in FIG. 7A.

[0066] FIG. 8 shows images of flow time series for olfactory delivery of technetium-99.

[0067] FIGs. 9A-9B are graphs showing particle size after processing by the RSH device and a nebulizer. (A) mRNA-LNP formulation. (B) saRNA-LNP formulation. Control refers to formulations not processed by either device.

[0068] FIGs. 10A-10B are graphs showing PDI (polydispersity index) after processing by the RSH device and a nebulizer. (A) mRNA-LNP formulation. (B) saRNA-LNP formulation. Control refers to formulations not processed by either device.Mintz Ref.: 062706-519001 WO

[0069] FIGs. 11A-11B are graphs showing raw readouts from Ribogreen assays for measuring encapsulation after processing by the RSH device and a nebulizer. (A) mRNA-LNP formulation. (B) saRNA-LNP formulation. Control refers to formulations not processed by either device.

[0070] FIGs. 12A-12B are graphs showing encapsulation efficiency after processing by the RSH device and a nebulizer. (A) mRNA-LNP formulation. (B) saRNA-LNP formulation. Control refers to formulations not processed by either device.

[0071] FIGs. 13A-13B are graphs showing zeta potential after processing by the RSH device and a nebulizer. (A) mRNA-LNP formulation. (B) saRNA-LNP formulation. Control refers to formulations not processed by either device.

[0072] FIGs. 14A-14D are graphs showing results of luciferase cell expression assays after processing by the RSH device and a nebulizer. (A) mRNA-LNP formulation. (B) saRNA-LNP formulation. Control refers to formulations not processed by either device. (C) and (D) are normalized data of (A) and (B), respectively.

[0073] FIG. 15 depicts device setup for delivery using a low-shear device (RSH device) according to some embodiments.

[0074] FIG. 16 depicts a device setup for spray delivery according to some embodiments.

[0075] FIGs. 17A-17B are graphs depicting bioluminescence levels post-dosing. (A) At the prone position. (B) At the supine position.

[0076] FIGs. 18A-18G depict representative images from whole-body bioluminescence imaging of the prone position or the supine position of mice at the specific time points post-dosing. (A) 1 hour post-dosing. (B) 3 hours post-dosing. (C) 6 hours post-dosing. (D) 8 hours post-dosing. (E) 24 hour post-dosing. (F) 48 hour post-dosing. (G) 72 hours post-dosing.DETAILED DESCRIPTION

[0077] This disclosure provides methods and devices for delivery of delivery vehicles to a target region of a nasal cavity, such as an olfactory region, of a subject.

[0078] This disclosure provides an introducer device for targeted delivery of a composition to a target region of a nasal cavity of a subject, which can be readily actuated and positioned by users of various patient populations, and reliably deliver the composition to the target region of the nasal cavity. Exemplary devices utilize a dual nostril inserter with a columella engaging portion positioned therebetween in order to quickly and reliably seat the insertable portions of the device within a target ejection zone of a subject’s nasal cavity. The exemplary devices disclosed herein can permit for quick,Mintz Ref.: 062706-519001 WO easy, and reliable positioning of a dispensing element within the nasal channel so as to permit for accurate and targeted deposition of compositions to target regions of the nasal cavity. Further, exemplary devices disclosed herein can enable quick, easy, and reliable positioning and targeted deposition across diverse patient populations (e.g., users that are elderly, cognitively impaired, dexterity impaired, or have variations in nasal anatomy), by users of varying skill (e.g., untrained bystanders), and under high stress circumstances (e.g., a medical emergency), which may otherwise prevent proper use of intranasal delivery devices.

[0079] The devices of the present disclosure may comprise housing comprising one or a combination of the following: a subject engaging portion, one or more insertable portions, one or more dispensing elements, and a trigger.

[0080] FIG. 1A depicts a bottom view of an exemplary embodiment of a subject’s nose. FIG. IB depicts a side view of an exemplary embodiment of a subject’s nose. In some embodiments the nose 1 has a columella region 10 between the entrance to two nasal channels 20. In some embodiments, the nose 1 has an external nasal valve 12 coupled to the nasal channel 20. In some embodiments, the nose 1 has an internal nasal valve 13 (INV) coupled to the nasal channel 20.

[0081] FIG. 1C depicts a side view of an exemplary embodiment of a subject’s nasal cavity showing an inferior turbinate 16, a middle turbinate 15, and a superior turbinate 14. FIG. ID depicts an angled side view of an exemplary embodiment of a representative subject’s nasal channels 20 from the vestibules 21 to the olfactory clefts 23 based on a posteriorly oriented plane 17 showing the middle turbinates 15. FIG. IE depicts an angled side view of an exemplary embodiment of a representative subject’s nasal channels 20 from the vestibules to the olfactory clefts based on an anteriorly oriented plane 18. FIG. IF depicts a side view of an exemplary embodiment of a representative subject’s target region 19. FIG. 1G depicts a front, base and perspective view of an exemplary embodiment of a representative subject’s nose exposing the columella region 10. The respiratory regions comprise turbinates that present physical obstacles to delivery to the upper reaches of a nasal channels 20, e.g., the olfactory clefts 23. Each respiratory region comprises at least one superior turbinate 14. Each respiratory region comprises at least one middle turbinate 15. Each respiratory region comprises at least one inferior turbinate 16. Each respiratory region comprises at least one posterior pathway 17 that involves at least one middle turbinate 15. Each respiratory region comprises at least one anterior pathway 18 that does not involve at least one middle turbinate 15.

[0082] In some cases, the middle turbinate 15 comprises a physical obstruction for composition delivery to an olfactory cleft. In some cases, the middle turbinate 15 comprises a most anterior aspectMintz Ref.: 062706-519001 WO about aligned with the cheek bone. In some embodiments, the middle turbinate 15 comprises a most anterior aspect not aligned with the cheek bone. In some cases, the nasal channels 20 simplify anteriorly, and comprise angled pathways without one or more turbinates presenting physical obstacles to delivering a composition 111 to the upper nasal channels, including the olfactory clefts 23, or directing compositions 111 down one or more meatuses, e.g., the middle meatus 30, to the nasopharynx. In some cases, the nasal channel 20 comprises one pathway from the vestibule 21 to the olfactory cleft 23 based on an anteriorly oriented plane with a target ejection point 19. In some embodiments, the nose comprises the nasal septum 24, upper lateral cartilage 25, and lower lateral cartilage 26.

[0083] FIG. 1H depicts a side view of an exemplary embodiment of a representative subject’s nasal cavity. In some cases, the subject’s nasal cavity 11 comprises the nasal vestibule 21, inferior turbinate 16, middle turbinate 15, superior turbinate 14, cribriform plate 31 or a combination thereof.

[0084] FIG. II depicts a side view of an exemplary embodiment of a representative subject’s sinus. In some cases, the subject’s sinus comprises the inferior turbinate 16, middle turbinate 15, superior turbinate 14, cribriform plate 31, middle meatus 30, or a combination thereof.

[0085] FIG. 1J depicts a side view of an exemplary embodiment of a representative subject’s nasal cavity. In some embodiments, the nasal cavity 11 comprises the nasal bone 33, septal-lateral cartilage junction 27, lower lateral cartilage 26, anterior nasal spine 32, or a combination thereof.I. Methods

[0086] It is demonstrated herein that lipid nanoparticles delivered by a proprietary low-shear device can result in superior quality of the lipid nanoparticles as compared to a high-shear device, such as a nebulizer.

[0087] Accordingly, in one aspect, provided herein is a method of delivering a delivery vehicle to a subject in need thereof, the method comprising: delivering a formulation comprising the delivery vehicle to a targeted region of a nasal cavity of the subject, wherein the formulation is delivered as a liquid jet. In one aspect, provided herein is a method of delivering a delivery vehicle to a subject in need thereof. The method can comprise delivering a formulation. The formulation can comprise the delivery vehicle. Delivering the formulation can comprise delivering to a targeted region. The targeted region can be a targeted region of a nasal cavity of the subject. Delivering the formulation can comprise delivering as a liquid jet.

[0088] The method can deliver at least 50% of the formulation to the target region.Mintz Ref.: 062706-519001 WO

[0089] In some embodiments, at least about 50% of the formulation is delivered to the target region of the subject. In some embodiments, at least about 55% of the formulation is delivered to the target region of the subject. In some embodiments, at least about 60% of the formulation is delivered to the target region of the subject. In some embodiments, at least about 65% of the formulation is delivered to the target region of the subject. In some embodiments, at least about 70% of the formulation is delivered to the target region of the subject. In some embodiments, at least about 75% of the formulation is delivered to the target region of the subject. In some embodiments, at least about 80% of the formulation is delivered to the target region of the subject. In some embodiments, at least about 85% of the formulation is delivered to the target region of the subject. In some embodiments, at least about 90% of the formulation is delivered to the target region of the subject. In some embodiments, at least about 95% of the formulation is delivered to the target region of the subject.

[0090] The formulation can be ejected from a dispensing element. In some embodiments, at least about 70% of the formulation can pass through a circular opening having a diameter of between about 5 mm and 6 mm placed about 25 mm from where the formulation is ejected from the dispensing element. In some embodiments, at least about 75% of the formulation can pass through a circular opening having a diameter of between about 5 mm and 6 mm placed about 25 mm from where the formulation is ejected from the dispensing element. In some embodiments, at least about 80% of the formulation can pass through a circular opening having a diameter of between about 5 mm and 6 mm placed about 25 mm from where the formulation is ejected from the dispensing element. In some embodiments, at least about 85% of the formulation can pass through a circular opening having a diameter of between about 5 mm and 6 mm placed about 25 mm from where the formulation is ejected from the dispensing element. In some embodiments, at least about 90% of the formulation can pass through a circular opening having a diameter of between about 5 mm and 6 mm placed about 25 mm from where the formulation is ejected from the dispensing element. In some embodiments, at least about 95% of the formulation can pass through a circular opening having a diameter of between about 5 mm and 6 mm placed about 25 mm from where the formulation is ejected from the dispensing element.

[0091] In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 100 cP. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 90 cP. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 80 cP. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 70 cP. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 60 cP. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 50 cP. In someMintz Ref.: 062706-519001 WO embodiments, the formulation has a viscosity of between about 0.5 cP and about 40 cP. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 30 cP. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 20 cP. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 10 cP. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 5 cP. In some embodiments, the formulation has a viscosity of between about 5 cP and about 10 cP. In some embodiments, the formulation has a viscosity of between about 10 cP and about 15 cP. In some embodiments, the formulation has a viscosity of between about 15 cP and about 20 cP. In some embodiments, the formulation has a viscosity of between about 20 cP and about 25 cP. In some embodiments, the formulation has a viscosity of between about 25 cP and about 30 cP. In some embodiments, the formulation has a viscosity of between about 30 cP and about 35 cP. In some embodiments, the formulation has a viscosity of between about 35 cP and about 40 cP. In some embodiments, the formulation has a viscosity of between about 40 cP and about 45 cP. In some embodiments, the formulation has a viscosity of between about 45 cP and about 50 cP. In some embodiments, the formulation has a viscosity of between about 50 cP and about 55 cP. In some embodiments, the formulation has a viscosity of between about 55 cP and about 60 cP. In some embodiments, the formulation has a viscosity of between about 60 cP and about 65 cP. In some embodiments, the formulation has a viscosity of between about 65 cP and about 70 cP. In some embodiments, the formulation has a viscosity of between about 70 cP and about 75 cP. In some embodiments, the formulation has a viscosity of between about 75 cP and about 80 cP. In some embodiments, the formulation has a viscosity of between about 85 cP and about 90 cP. In some embodiments, the formulation has a viscosity of between about )5 cP and about 100 cP. In some embodiments, the formulation has a viscosity of higher than 100 cP. In some embodiments, the formulation has a viscosity of between about 45 cP and about 55 cP. In some embodiments, the formulation has a viscosity of between about 48 cP and about 52 cP. In some embodiments, the formulation has a viscosity of about 1 cP. In some embodiments, the formulation has a viscosity of about 50 cP.

[0092] In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 30 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 25 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 20 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity ofMintz Ref.: 062706-519001 WO between about 1 m / s and about 15 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 10 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 5 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 2 m / s and about 30 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 2 m / s and about 25 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 2 m / s and about 20 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 2 m / s and about 15 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 2 m / s and about 10 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 2 m / s and about 5 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 3 m / s and about 30 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 3 m / s and about 25 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 3 m / s and about 20 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 3 m / s and about 15 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 3 m / s and about 10 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 3 m / s and about 5 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 4 m / s and about 30 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 4 m / s and about 25 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 4 m / s and about 20 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 4 m / s and about 15 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 4 m / s and about 10 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 4 m / s and about 5 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 5 m / s and about 30 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 5 m / s and about 25 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 5 m / s and about 20 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity ofMintz Ref.: 062706-519001 WO between about 5 m / s and about 15 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 5 m / s and about 10 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 10 m / s and about 30 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 10 m / s and about 25 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 10 m / s and about 20 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 10 m / s and about 15 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 15 m / s and about 30 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 15 m / s and about 25 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 15 m / s and about 20 m / s. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 20 m / s and about 30 m / s.

[0093] In some embodiments, the formulation is ejected from the dispensing element at a velocity. In some embodiments, the velocity is about 1 m / s. In some embodiments, the velocity is about 2 m / s. In some embodiments, the velocity is about 3 m / s. In some embodiments, the velocity is about 4 m / s. In some embodiments, the velocity is about 5 m / s. In some embodiments, the velocity is about 6 m / s. In some embodiments, the velocity is about 7 m / s. In some embodiments, the velocity is about 8 m / s. In some embodiments, the velocity is about 9 m / s. In some embodiments, the velocity is about 10 m / s. In some embodiments, the velocity is about 11 m / s. In some embodiments, the velocity is about 12 m / s. In some embodiments, the velocity is about 13 m / s. In some embodiments, the velocity is about 14 m / s. In some embodiments, the velocity is about 15 m / s. In some embodiments, the velocity is about 16 m / s. In some embodiments, the velocity is about 17 m / s. In some embodiments, the velocity is about 18 m / s. In some embodiments, the velocity is about 19 m / s. In some embodiments, the velocity is about 20 m / s. In some embodiments, the velocity is about 21 m / s. In some embodiments, the velocity is about 22 m / s. In some embodiments, the velocity is about 23 m / s. In some embodiments, the velocity is about 24 m / s. In some embodiments, the velocity is about 25 m / s. In some embodiments, the velocity is about 26 m / s. In some embodiments, the velocity is about 27 m / s. In some embodiments, the velocity is about 28 m / s. In some embodiments, the velocity is about 29 m / s. In some embodiments, the velocity is about 30 m / s. In some embodiments, the velocity is between about 1 m / s and about 4 m / s. In some embodiments, the velocity is between about 2 m / s and about 5 m / s. In some embodiments, the velocity is between about 3 m / s and about 6 m / s. In some embodiments, the velocity is between about 4 m / sMintz Ref.: 062706-519001 WO and about 7 m / s. In some embodiments, the velocity is between about 5 m / s and about 8 m / s. In some embodiments, the velocity is between about 6 m / s and about 9 m / s. In some embodiments, the velocity is between about 7 m / s and about 10 m / s. In some embodiments, the velocity is between about 8 m / s and about 1' m / s. In some embodiments, the velocity is between about 9 m / s and about 11 m / s. In some embodiments, the velocity is between about 10 m / s and about 12 m / s. In some embodiments, the velocity is between about 11 m / s and about 14 m / s. In some embodiments, the velocity is between about 12 m / s and about 15 m / s. In some embodiments, the velocity is between about 13 m / s and about 16 m / s. In some embodiments, the velocity is between about 14 m / s and about 17 m / s. In some embodiments, the velocity is between about 15 m / s and about 18 m / s. In some embodiments, the velocity is between about 16 m / s and about 19 m / s. In some embodiments, the velocity is between about 17 m / s and about 20 m / s. In some embodiments, the velocity is between about 17 m / s and about 20 m / s. In some embodiments, the velocity is between about 18 m / s and about 21 m / s. In some embodiments, the velocity is between about 19 m / s and about 22 m / s. In some embodiments, the velocity is between about 20 m / s and about 23 m / s. In some embodiments, the velocity is between about 21 m / s and about 24 m / s. In some embodiments, the velocity is between about 22 m / s and about 25 m / s. In some embodiments, the velocity is between about 23 m / s and about 26 m / s. In some embodiments, the velocity is between about 24 m / s and about 27 m / s. In some embodiments, the velocity is between about 25 m / s and about 28 m / s. In some embodiments, the velocity is between about 26 m / s and about 29 m / s. In some embodiments, the velocity is between about 27 m / s and about 30 m / s.

[0094] In some embodiments, the formulation is ejected from a dispensing element, wherein at least about 75% of the formulation passes through a circular opening having a diameter of between about 5 mm and about 6 mm placed about 25 mm from where the formulation is ejected from the dispensing element. In some embodiments, the formulation has a viscosity of about 1 cP and is ejected from the dispensing element at a velocity of between about 2 m / s and about 4 m / s. In some embodiments, the formulation has a viscosity of about 50 cP and is ejected from the dispensing element at a velocity of between about 25 m / s and about 30 m / s. In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 50 cP. In some embodiments, the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 30 m / s.

[0095] The formulation can be delivered as a liquid jet or laminar flow. The formulation can be delivered as a flow having a Reynold’s number of 2300 or less.Mintz Ref.: 062706-519001 WO

[0096] Laminar flow can refer to a smooth, regular movement of fluids where adjacent layers of particles experience relatively little mixing.

[0097] In some embodiments, the formulation is delivered as a liquid jet.

[0098] In some embodiments, the formulation is delivered as a laminar flow.

[0099] In some embodiments, delivering the formulation as a laminar flow increases on target delivery of the composition to the target region compared to delivering the formulation with a spray ejection profile. In some embodiments, delivering the formulation as a laminar flow decreases off target delivery of the composition to the nasal cavity compared to delivering the formulation with a spray ejection profile. In some embodiments, delivering the formulation as a laminar flow increases on target delivery of the composition to the target region and decreases off target delivery of the composition to the nasal cavity compared to delivering the formulation with a spray ejection profile.

[0100] In some embodiments, the formulation is delivered as a liquid jet.

[0101] In some embodiments, delivering the formulation as a liquid jet increases on target delivery of the composition to the target region compared to delivering the formulation with a spray ejection profile. In some embodiments, delivering the formulation as a liquid jet decreases off target delivery of the composition to the nasal cavity compared to delivering the formulation with a spray ejection profile. In some embodiments, delivering the formulation as a liquid jet increases on target delivery of the composition to the target region and decreases off target delivery of the composition to the nasal cavity compared to delivering the formulation with a spray ejection profile.

[0102] In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 2300 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 2200 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 2100 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 2000 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 1900 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 1800 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 1700 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 1600 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 1500 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 1400 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 1300 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 1200 or less. In someMintz Ref.: 062706-519001 WO embodiments, the formulation is delivered as a flow having a Reynold’s number of 1100 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 1000 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 900 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 800 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 700 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 600 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 500 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 400 or less. In some embodiments, the formulation is delivered as a flow having a Reynold’s number of 300 or less.

[0103] In some embodiments, delivering the formulation as a flow having a Reynold’s number of 2300 or less increases on target delivery of the composition to the target region compared to delivering the formulation with a spray ejection profile. In some embodiments, delivering the formulation as a flow having a Reynold’s number of 2300 or less decreases off target delivery of the composition to the nasal cavity compared to delivering the formulation with a spray ejection profile. In some embodiments, delivering the formulation as a flow having a Reynold’s number of 2300 or less increases on target delivery of the composition to the target region and decreases off target delivery of the composition to the nasal cavity compared to delivering the formulation with a spray ejection profile.

[0104] The target region can be a target region in a nasal cavity of the subject. For example, the target region can be an olfactory region, a turbinate region, a nasopharynx region, a nasal associated lymphatic tissue, or any combination thereof.

[0105] In some embodiments, the target region comprises an olfactory region. In some embodiments, the target region comprises a turbinate region. In some embodiments, the target region comprises a middle turbinate region. In some embodiments, the target region comprises a superior turbinate region. In some embodiments, the target region comprises an inferior turbinate region. In some embodiments, the target region comprises a nasopharynx region. In some embodiments, the target region comprises a nasal-associated lymphatic tissue.

[0106] In some embodiments, selectively delivering the formulation to the target region of the subject comprises inserting a dispensing element into an ejection zone of a nasal cavity of the subject.

[0107] In some embodiments, the method further comprises inserting a dispensing element into a nasal cavity of the subject.Mintz Ref.: 062706-519001 WO

[0108] In some embodiments, the dispensing element is positioned from about 0.1 mm to about 30 mm from the olfactory region or an anterior entry to the olfactory region. In some embodiments, the tip is positioned from about 0.1 mm to about 25 mm from the olfactory region or an anterior entry to the olfactory region. In some embodiments, the tip is positioned from about 0.1 mm to about 3 mm, about 0.1 mm to about 5 mm, about 0.1 mm to about 9 mm, about 0.1 mm to about 12 mm, about 0.1 mm to about 15 mm, about 0.1 mm to about 18 mm, about 0.1 mm to about 20 mm, about 0.1 mm to about 25 mm, about 0.1 mm to about 30 mm, about 3 mm to about 5 mm, about 3 mm to about 9 mm, about 3 mm to about 12 mm, about 3 mm to about 15 mm, about 3 mm to about 18 mm, about 3 mm to about 20 mm, about 3 mm to about 25 mm, about 3 mm to about 30 mm, about 5 mm to about 9 mm, about 5 mm to about 12 mm, about 5 mm to about 15 mm, about 5 mm to about 18 mm, about 5 mm to about 20 mm, about 5 mm to about 25 mm, about 5 mm to about 30 mm, about 9 mm to about 12 mm, about 9 mm to about 15 mm, about 9 mm to about 18 mm, about 9 mm to about 20 mm, about 9 mm to about 25 mm, about 9 mm to about 30 mm, about 12 mm to about 15 mm, about 12 mm to about 18 mm, about 12 mm to about 20 mm, about 12 mm to about 25 mm, about 12 mm to about 30 mm, about 15 mm to about 18 mm, about 15 mm to about 20 mm, about 15 mm to about 25 mm, about 15 mm to about 30 mm, about 18 mm to about 20 mm, about 18 mm to about 25 mm, about 18 mm to about 30 mm, about 20 mm to about 25 mm, about 20 mm to about 30 mm, or about 25 mm to about 30 mm, including increments therein. In some embodiments, the tip is positioned from about 0.1 mm, about 3 mm, about 5 mm, about 9 mm, about 12 mm, about 15 mm, about 18 mm, about 20 mm, about 25 mm, or about 30 mm from the olfactory region or an anterior entry to the olfactory region. In some embodiments, the tip is positioned from at least about 0.1 mm, about 3 mm, about 5 mm, about 9 mm, about 12 mm, about 15 mm, about 18 mm, about 20 mm, or about 25 mm from the olfactory region or an anterior entry to the olfactory region. In some embodiments, the dispensing element is positioned from at most about 3 mm, about 5 mm, about 9 mm, about 12 mm, about 15 mm, about 18 mm, about 20 mm, about 25 mm, or about 30 mm from the olfactory region or an anterior entry to the olfactory region.

[0109] In some embodiments, the method further comprises ejecting the formulation from an ejection zone of the nasal cavity.

[0110] Ejection Zone

[0111] In some embodiments, dispensing the formulation from the ejection zone increases on target delivery of the composition to the target region compared to dispensing the formulation outside the ejection zone. In some embodiments, dispensing the formulation from the ejection zone decreases offMintz Ref.: 062706-519001 WO target delivery of the composition to the nasal cavity compared to dispensing the formulation outside the ejection zone. In some embodiments, dispensing the formulation from the ejection zone increases on target delivery of the composition to the target region and decreases off target delivery of the composition to the nasal cavity compared to dispensing the formulation outside the ejection zone.

[0112] Referring to FIGs. 4A-4J. In some embodiments, the ejection zone 29 is: 0mm to 30mm superior to a horizontal line 627 that intersects the anterior aspect of the internal nasal valve 13, and 0mm to 20mm anterior to an inclined line 620 that intersects the anterior aspect of the middle turbinate 15 and the posterior aspect of the vestibule 21.

[0113] In some embodiments, the ejection zone 29 is further 0 mm to 40mm inferior to a horizontal line 621 that is parallel to the inferior aspect of the olfactory cleft 23, 0mm to 20mm posterior to the internal nasal dorsum 622, 10mm to 50mm superior to a horizontal line 617 that intersects the inferior aspect of the columella 10, 0mm to 30mm superior to a horizontal line 627 that intersects the superior aspect of the inferior turbinate 16, 0mm to 3mm from the septum 24 , or any combination thereof.

[0114] In some embodiments, the ejection zone 29 is a trapezium or irregular quadrilateral comprising (i) an inferior side 29A being a 10-25mm line extending posteriorly and horizontally from the anterior aspect of the internal nasal valve 13, (ii) an anterior side 29B being a 10-35mm line extending superiorly and parallel to the internal nasal dorsum 622 from the anterior aspect of the internal nasal valve 13, (iii) a superior side 29C being a 10-25mm line extending posteriorly and horizontally from a point on the internal nasal dorsum 622 that is 0-10mm inferior to the inferior aspect of the olfactory cleft 23, and (iv) a posterior line 29D being a 10-35mm line extending vertically along a plane that intersects the anterior aspect of the middle turbinate 15.

[0115] In some embodiments, dispensing the formulation from the ejection zone 29 increases on target deposition of the formulation to an olfactory cleft 23.

[0116] In some embodiments, the subject engaging portion 106 engages a columella region 10 of the subject to seat a distal end 128 of the insertable portion 107 within an ejection zone 29 of a nasal cavity 11 of the subject, wherein the ejection zone 29 is: 0mm to 30mm superior to a horizontal line 627 that intersects the anterior aspect of the internal nasal valve 13, and 0mm to 20mm anterior to an inclined line 620 that intersects the anterior aspect of the middle turbinate 15 and the posterior aspect of the vestibule 21, and one or more of the following: 0mm to 40mm inferior to a horizontal line 621 that is parallel to the inferior aspect of the olfactory cleft 23, 0mm to 20mm posterior to the internal nasal dorsum 622, 10mm to 50mm superior to a horizontal line 623 that intersects the inferior aspectMintz Ref.: 062706-519001 WO of the columella 10, 0mm to 30mm superior to a horizontal line 627 that intersects the superior aspect of the inferior turbinate 16, 0mm to 3mm from the septum 24 or any combination thereof.

[0117] In some embodiments, the ejection zone is superior to a horizontal line that intersects the anterior aspect of the internal nasal valve by about 1 mm to about 30 mm. In some embodiments, the ejection zone is superior to a horizontal line that intersects the anterior aspect of the internal nasal valve by about 1 mm to about 2 mm, about 1 mm to about 4 mm, about 1 mm to about 6 mm, about 1 mm to about 8 mm, about 1 mm to about 10 mm, about 1 mm to about 14 mm, about 1 mm to about 18 mm, about 1 mm to about 22 mm, about 1 mm to about 26 mm, about 1 mm to about 30 mm, about 2 mm to about 4 mm, about 2 mm to about 6 mm, about 2 mm to about 8 mm, about 2 mm to about 10 mm, about 2 mm to about 14 mm, about 2 mm to about 18 mm, about 2 mm to about 22 mm, about 2 mm to about 26 mm, about 2 mm to about 30 mm, about 4 mm to about 6 mm, about 4 mm to about 8 mm, about 4 mm to about 10 mm, about 4 mm to about 14 mm, about 4 mm to about 18 mm, about 4 mm to about 22 mm, about 4 mm to about 26 mm, about 4 mm to about 30 mm, about 6 mm to about 8 mm, about 6 mm to about 10 mm, about 6 mm to about 14 mm, about 6 mm to about 18 mm, about 6 mm to about 22 mm, about 6 mm to about 26 mm, about 6 mm to about 30 mm, about 8 mm to about 10 mm, about 8 mm to about 14 mm, about 8 mm to about 18 mm, about 8 mm to about 22 mm, about 8 mm to about 26 mm, about 8 mm to about 30 mm, about 10 mm to about 14 mm, about 10 mm to about 18 mm, about 10 mm to about 22 mm, about 10 mm to about 26 mm, about 10 mm to about 30 mm, about 14 mm to about 18 mm, about 14 mm to about 22 mm, about 14 mm to about 26 mm, about 14 mm to about 30 mm, about 18 mm to about 22 mm, about 18 mm to about 26 mm, about 18 mm to about 30 mm, about 22 mm to about 26 mm, about 22 mm to about 30 mm, or about 26 mm to about 30 mm, including increments therein. In some embodiments, the ejection zone is superior to a horizontal line that intersects the anterior aspect of the internal nasal valve by about 1 mm, about 2 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 14 mm, about 18 mm, about 22 mm, about 26 mm, or about 30 mm.

[0118] In some embodiments, the ejection zone is superior to a horizontal line that intersects the anterior aspect of the internal nasal valve by at least about 1 mm, about 2 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 14 mm, about 18 mm, about 22 mm, or about 26 mm. In some embodiments, the ejection zone is superior to a horizontal line that intersects the anterior aspect of the internal nasal valve by at most about 2 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 14 mm, about 18 mm, about 22 mm, about 26 mm, or about 30 mm. In some embodiments, the ejection zone is anterior to an inclined line that intersects the anterior aspect of the middle turbinateMintz Ref.: 062706-519001 WO and the posterior aspect of the vestibule by about 1 mm to about 20 mm. In some embodiments, the ejection zone is anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule by about 1 mm to about 2 mm, about 1 mm to about 4 mm, about 1 mm to about 6 mm, about 1 mm to about 8 mm, about 1 mm to about 10 mm, about 1 mm to about 12 mm, about 1 mm to about 14 mm, about 1 mm to about 16 mm, about 1 mm to about 18 mm, about 1 mm to about 20 mm, about 2 mm to about 4 mm, about 2 mm to about 6 mm, about 2 mm to about 8 mm, about 2 mm to about 10 mm, about 2 mm to about 12 mm, about 2 mm to about 14 mm, about 2 mm to about 16 mm, about 2 mm to about 18 mm, about 2 mm to about 20 mm, about 4 mm to about 6 mm, about 4 mm to about 8 mm, about 4 mm to about 10 mm, about 4 mm to about 12 mm, about 4 mm to about 14 mm, about 4 mm to about 16 mm, about 4 mm to about 18 mm, about 4 mm to about 20 mm, about 6 mm to about 8 mm, about 6 mm to about 10 mm, about 6 mm to about 12 mm, about 6 mm to about 14 mm, about 6 mm to about 16 mm, about 6 mm to about 18 mm, about 6 mm to about 20 mm, about 8 mm to about 10 mm, about 8 mm to about 12 mm, about 8 mm to about 14 mm, about 8 mm to about 16 mm, about 8 mm to about 18 mm, about 8 mm to about 20 mm, about 10 mm to about 12 mm, about 10 mm to about 14 mm, about 10 mm to about 16 mm, about 10 mm to about 18 mm, about 10 mm to about 20 mm, about 12 mm to about 14 mm, about 12 mm to about 16 mm, about 12 mm to about 18 mm, about 12 mm to about 20 mm, about 14 mm to about 16 mm, about 14 mm to about 18 mm, about 14 mm to about 20 mm, about 16 mm to about 18 mm, about 16 mm to about 20 mm, or about 18 mm to about 20 mm, including increments therein.

[0119] In some embodiments, the ejection zone is anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule by about 1 mm, about 2 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 12 mm, about 14 mm, about 16 mm, about 18 mm, or about 20 mm. In some embodiments, the ejection zone is anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule by at least about 1 mm, about 2 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 12 mm, about 14 mm, about 16 mm, or about 18 mm. In some embodiments, the ejection zone is anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule by at most about 2 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 12 mm, about 14 mm, about 16 mm, about 18 mm, or about 20 mm.

[0120] In some embodiments, the ejection zone is inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft by about 1 mm to about 35 mm. In some embodiments, the ejection zone is inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft by aboutMintz Ref.: 062706-519001 WO1 mm to about 2 mm, about 1 mm to about 3 mm, about 1 mm to about 4 mm, about 1 mm to about 5 mm, about 1 mm to about 8 mm, about 1 mm to about 12 mm, about 1 mm to about 15 mm, about 1 mm to about 18 mm, about 1 mm to about 21 mm, about 1 mm to about 25 mm, about 1 mm to about 30 mm, about 1 mm to about 35 mm, about 2 mm to about 3 mm, about 2 mm to about 4 mm, about 2 mm to about 5 mm, about 2 mm to about 8 mm, about 2 mm to about 12 mm, about 2 mm to about 15 mm, about 2 mm to about 18 mm, about 2 mm to about 21 mm, about 2 mm to about 25 mm, about 2 mm to about 30 mm, about 2 mm to about 35 mm, about 3 mm to about 4 mm, about 3 mm to about 5 mm, about 3 mm to about 8 mm, about 3 mm to about 12 mm, about 3 mm to about 15 mm, about 3 mm to about 18 mm, about 3 mm to about 21 mm, about 3 mm to about 25 mm, about 3 mm to about 30 mm, about 3 mm to about 35 mm, about 4 mm to about 5 mm, about 4 mm to about 8 mm, about 4 mm to about 12 mm, about 4 mm to about 15 mm, about 4 mm to about 18 mm, about 4 mm to about 21 mm, about 4 mm to about 25 mm, about 4 mm to about 30 mm, about 4 mm to about 35 mm, about 5 mm to about 8 mm, about 5 mm to about 12 mm, about 5 mm to about 15 mm, about 5 mm to about 18 mm, about 5 mm to about 21 mm, about 5 mm to about 25 mm, about 5 mm to about 30 mm, about 5 mm to about 35 mm, about 8 mm to about 15 mm, about 8 mm to about 18 mm, about 8 mm to about 21 mm, about 8 mm to about 25 mm, about 8 mm to about 30 mm, about 8 mm to about 35 mm, about 12 mm to about 15 mm, about 12 mm to about 18 mm, about 12 mm to about 21 mm, about 12 mm to about 25 mm, about 12 mm to about 30 mm, about 12 mm to about 35 mm, about 15 mm to about 18 mm, about 15 mm to about 21 mm, about 15 mm to about 25 mm, about 15 mm to about 30 mm, about 15 mm to about 35 mm, about 18 mm to about 21 mm, about 18 mm to about 25 mm, about 18 mm to about 30 mm, about 18 mm to about 35 mm, about 21 mm to about 25 mm, about 21 mm to about 30 mm, about 21 mm to about 35 mm, about 25 mm to about 30 mm, about 25 mm to about 35 mm, or about 30 mm to about 35 mm, including increments therein. In some embodiments, the ejection zone is inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft by about 1 mm, about 2 mm, about 3 mm, about 4 mm, about 5 mm, about 8 mm, about 12 mm, about 15 mm, about 18 mm, about 21 mm, about 25 mm, about 30 mm, or about 35 mm. In some embodiments, the ejection zone is inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft by at least about 1 mm, about 2 mm, about 3 mm, about 4 mm, about 5 mm, about 8 mm, about 12 mm, about 15 mm, about 18 mm, about 21 mm, about 25 mm, or about 30 mm. In some embodiments, the ejection zone is inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft by at most about 2 mm, about 3 mm, about 4 mm, about 5 mm, about 8 mm, about 12 mm, about 15 mm, about 18 mm, about 21 mm, about 25 mm, about 30 mm, or about 35 mm.Mintz Ref.: 062706-519001 WO

[0121] In some embodiments, the ejection zone is posterior to the internal nasal dorsum by about 1 mm to about 20 mm. In some embodiments, the ejection zone is posterior to the internal nasal dorsum by about 1 mm to about 2 mm, about 1 mm to about 3 mm, about 1 mm to about 4 mm, about 1 mm to about 6 mm, about 1 mm to about 8 mm, about 1 mm to about 10 mm, about 1 mm to about 12 mm, about 1 mm to about 14 mm, about 1 mm to about 16 mm, about 1 mm to about 18 mm, about 1 mm to about 20 mm, about 2 mm to about 3 mm, about 2 mm to about 4 mm, about 2 mm to about 6 mm, about 2 mm to about 8 mm, about 2 mm to about 10 mm, about 2 mm to about 12 mm, about 2 mm to about 14 mm, about 2 mm to about 16 mm, about 2 mm to about 18 mm, about 2 mm to about 20 mm, about 3 mm to about 4 mm, about 3 mm to about 6 mm, about 3 mm to about 8 mm, about 3 mm to about 10 mm, about 3 mm to about 12 mm, about 3 mm to about 14 mm, about 3 mm to about 16 mm, about 3 mm to about 18 mm, about 3 mm to about 20 mm, about 4 mm to about 6 mm, about 4 mm to about 8 mm, about 4 mm to about 10 mm, about 4 mm to about 12 mm, about 4 mm to about 14 mm, about 4 mm to about 16 mm, about 4 mm to about 18 mm, about 4 mm to about 20 mm, about 6 mm to about 8 mm, about 6 mm to about 10 mm, about 6 mm to about 12 mm, about 6 mm to about 14 mm, about 6 mm to about 16 mm, about 6 mm to about 18 mm, about 6 mm to about 20 mm, about 8 mm to about 10 mm, about 8 mm to about 12 mm, about 8 mm to about 14 mm, about 8 mm to about 16 mm, about 8 mm to about 18 mm, about 8 mm to about 20 mm, about 10 mm to about 12 mm, about 10 mm to about 14 mm, about 10 mm to about 16 mm, about 10 mm to about 18 mm, about 10 mm to about 20 mm, about 12 mm to about 14 mm, about 12 mm to about 16 mm, about 12 mm to about 18 mm, about 12 mm to about 20 mm, about 14 mm to about 16 mm, about 14 mm to about 18 mm, about 14 mm to about 20 mm, about 16 mm to about 18 mm, about 16 mm to about 20 mm, or about 18 mm to about 20 mm, including increments therein. In some embodiments, the ejection zone is posterior to the internal nasal dorsum by about 1 mm, about 2 mm, about 3 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 12 mm, about 14 mm, about 16 mm, about 18 mm, or about 20 mm. In some embodiments, the ejection zone is posterior to the internal nasal dorsum by at least about 1 mm, about 2 mm, about 3 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 12 mm, about 14 mm, about 16 mm, or about 18 mm. In some embodiments, the ejection zone is posterior to the internal nasal dorsum by at most about 2 mm, about 3 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 12 mm, about 14 mm, about 16 mm, about 18 mm, or about 20 mm.

[0122] In some embodiments, the ejection zone is superior to a horizontal line that intersects the inferior aspect of the columella by about 10 mm to about 50 mm. In some embodiments, the ejection zone is superior to a horizontal line that intersects the inferior aspect of the columella by about 10 mmMintz Ref.: 062706-519001 WO to about 12 mm, about 10 mm to about 16 mm, about 10 mm to about 20 mm, about 10 mm to about 24 mm, about 10 mm to about 28 mm, about 10 mm to about 32 mm, about 10 mm to about 36 mm, about 10 mm to about 40 mm, about 10 mm to about 45 mm, about 10 mm to about 50 mm, about 12 mm to about 16 mm, about 12 mm to about 20 mm, about 12 mm to about 24 mm, about 12 mm to about 28 mm, about 12 mm to about 32 mm, about 12 mm to about 36 mm, about 12 mm to about 40 mm, about 12 mm to about 45 mm, about 12 mm to about 50 mm, about 16 mm to about 20 mm, about 16 mm to about 24 mm, about 16 mm to about 28 mm, about 16 mm to about 32 mm, about 16 mm to about 36 mm, about 16 mm to about 40 mm, about 16 mm to about 45 mm, about 16 mm to about 50 mm, about 20 mm to about 24 mm, about 20 mm to about 28 mm, about 20 mm to about 32 mm, about 20 mm to about 36 mm, about 20 mm to about 40 mm, about 20 mm to about 45 mm, about 20 mm to about 50 mm, about 24 mm to about 28 mm, about 24 mm to about 32 mm, about 24 mm to about 36 mm, about 24 mm to about 40 mm, about 24 mm to about 45 mm, about 24 mm to about 50 mm, about 28 mm to about 32 mm, about 28 mm to about 36 mm, about 28 mm to about 40 mm, about 28 mm to about 45 mm, about 28 mm to about 50 mm, about 32 mm to about 36 mm, about 32 mm to about 40 mm, about 32 mm to about 45 mm, about 32 mm to about 50 mm, about 36 mm to about 40 mm, about 36 mm to about 45 mm, about 36 mm to about 50 mm, about 40 mm to about 45 mm, about 40 mm to about 50 mm, or about 45 mm to about 50 mm, including increments therein. In some embodiments, the ejection zone is superior to a horizontal line that intersects the inferior aspect of the columella by about 10 mm, about 12 mm, about 16 mm, about 20 mm, about 24 mm, about 28 mm, about 32 mm, about 36 mm, about 40 mm, about 45 mm, or about 50 mm. In some embodiments, the ejection zone is superior to a horizontal line that intersects the inferior aspect of the columella by at least about 10 mm, about 12 mm, about 16 mm, about 20 mm, about 24 mm, about 28 mm, about 32 mm, about 36 mm, about 40 mm, or about 45 mm. In some embodiments, the ejection zone is superior to a horizontal line that intersects the inferior aspect of the columella by at most about 12 mm, about 16 mm, about 20 mm, about 24 mm, about 28 mm, about 32 mm, about 36 mm, about 40 mm, about 45 mm, or about 50 mm.

[0123] In some embodiments, the ejection zone is superior to a horizontal line that intersects the superior aspect of the inferior turbinate by about 1 mm to about 30 mm. In some embodiments, the ejection zone is superior to a horizontal line that intersects the superior aspect of the inferior turbinate by about 1 mm to about 2 mm, about 1 mm to about 4 mm, about 1 mm to about 6 mm, about 1 mm to about 8 mm, about 1 mm to about 10 mm, about 1 mm to about 12 mm, about 1 mm to about 15 mm, about 1 mm to about 18 mm, about 1 mm to about 21 mm, about 1 mm to about 25 mm, about 1Mintz Ref.: 062706-519001 WO mm to about 30 mm, about 2 mm to about 4 mm, about 2 mm to about 6 mm, about 2 mm to about 8 mm, about 2 mm to about 10 mm, about 2 mm to about 12 mm, about 2 mm to about 15 mm, about 2 mm to about 18 mm, about 2 mm to about 21 mm, about 2 mm to about 25 mm, about 2 mm to about 30 mm, about 4 mm to about 6 mm, about 4 mm to about 8 mm, about 4 mm to about 10 mm, about 4 mm to about 12 mm, about 4 mm to about 15 mm, about 4 mm to about 18 mm, about 4 mm to about 21 mm, about 4 mm to about 25 mm, about 4 mm to about 30 mm, about 6 mm to about 8 mm, about 6 mm to about 10 mm, about 6 mm to about 12 mm, about 6 mm to about 15 mm, about 6 mm to about 18 mm, about 6 mm to about 21 mm, about 6 mm to about 25 mm, about 6 mm to about 30 mm, about 8 mm to about 10 mm, about 8 mm to about 12 mm, about 8 mm to about 15 mm, about 8 mm to about 18 mm, about 8 mm to about 21 mm, about 8 mm to about 25 mm, about 8 mm to about 30 mm, about 10 mm to about 12 mm, about 10 mm to about 15 mm, about 10 mm to about 18 mm, about 10 mm to about 21 mm, about 10 mm to about 25 mm, about 10 mm to about 30 mm, about 12 mm to about 15 mm, about 12 mm to about 18 mm, about 12 mm to about 21 mm, about 12 mm to about 25 mm, about 12 mm to about 30 mm, about 15 mm to about 18 mm, about 15 mm to about 21 mm, about 15 mm to about 25 mm, about 15 mm to about 30 mm, about 18 mm to about 21 mm, about 18 mm to about 25 mm, about 18 mm to about 30 mm, about 21 mm to about 25 mm, about 21 mm to about 30 mm, or about 25 mm to about 30 mm, including increments therein. In some embodiments, the ejection zone is superior to a horizontal line that intersects the superior aspect of the inferior turbinate by about 1 mm, about 2 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 12 mm, about 15 mm, about 18 mm, about 21 mm, about 25 mm, or about 30 mm. In some embodiments, the ejection zone is superior to a horizontal line that intersects the superior aspect of the inferior turbinate by at least about 1 mm, about 2 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 12 mm, about 15 mm, about 18 mm, about 21 mm, or about 25 mm. In some embodiments, the ejection zone is superior to a horizontal line that intersects the superior aspect of the inferior turbinate by at most about 2 mm, about 4 mm, about 6 mm, about 8 mm, about 10 mm, about 12 mm, about 15 mm, about 18 mm, about 21 mm, about 25 mm, or about 30 mm. In some embodiments, the ejection zone is away from the septum by about 0.1 mm to about 3 mm. In some embodiments, the ejection zone is away from the septum by about 0.1 mm to about 0.2 mm, about 0.1 mm to about 0.4 mm, about 0.1 mm to about 0.6 mm, about 0.1 mm to about 0.8 mm, about 0.1 mm to about 1 mm, about 0.1 mm to about 1.25 mm, about 0.1 mm to about 1.5 mm, about 0.1 mm to about 1.75 mm, about 0.1 mm to about 2 mm, about 0.1 mm to about 2.5 mm, about 0.1 mm to about 3 mm, about 0.2 mm to about 0.4 mm, about 0.2 mm to about 0.6 mm, about 0.2 mm to about 0.8 mm, about 0.2 mm to about 1 mm, aboutMintz Ref.: 062706-519001 WO0.2 mm to about 1.25 mm, about 0.2 mm to about 1.5 mm, about 0.2 mm to about 1.75 mm, about 0.2 mm to about 2 mm, about 0.2 mm to about 2.5 mm, about 0.2 mm to about 3 mm, about 0.4 mm to about 0.6 mm, about 0.4 mm to about 0.8 mm, about 0.4 mm to about 1 mm, about 0.4 mm to about 1.25 mm, about 0.4 mm to about 1.5 mm, about 0.4 mm to about 1.75 mm, about 0.4 mm to about 2 mm, about 0.4 mm to about 2.5 mm, about 0.4 mm to about 3 mm, about 0.6 mm to about 0.8 mm, about 0.6 mm to about 1 mm, about 0.6 mm to about 1.25 mm, about 0.6 mm to about 1.5 mm, about 0.6 mm to about 1.75 mm, about 0.6 mm to about 2 mm, about 0.6 mm to about 2.5 mm, about 0.6 mm to about 3 mm, about 0.8 mm to about 1 mm, about 0.8 mm to about 1.25 mm, about 0.8 mm to about 1.5 mm, about 0.8 mm to about 1.75 mm, about 0.8 mm to about 2 mm, about 0.8 mm to about2.5 mm, about 0.8 mm to about 3 mm, about 1 mm to about 1.25 mm, about 1 mm to about 1.5 mm, about 1 mm to about 1.75 mm, about 1 mm to about 2 mm, about 1 mm to about 2.5 mm, about 1 mm to about 3 mm, about 1.25 mm to about 1.5 mm, about 1.25 mm to about 1.75 mm, about 1.25 mm to about 2 mm, about 1.25 mm to about 2.5 mm, about 1.25 mm to about 3 mm, about 1.5 mm to about 1.75 mm, about 1.5 mm to about 2 mm, about 1.5 mm to about 2.5 mm, about 1.5 mm to about 3 mm, about 1.75 mm to about 2 mm, about 1.75 mm to about 2.5 mm, about 1.75 mm to about 3 mm, about 2 mm to about 2.5 mm, about 2 mm to about 3 mm, or about 2.5 mm to about 3 mm, including increments therein. In some embodiments, the ejection zone is away from the septum by about 0.1 mm, about 0.2 mm, about 0.4 mm, about 0.6 mm, about 0.8 mm, about 1 mm, about 1.25 mm, about1.5 mm, about 1.75 mm, about 2 mm, about 2.5 mm, or about 3 mm. In some embodiments, the ejection zone is away from the septum by at least about 0.1 mm, about 0.2 mm, about 0.4 mm, about 0.6 mm, about 0.8 mm, about 1 mm, about 1.25 mm, about 1.5 mm, about 1.75 mm, about 2 mm, or about 2.5 mm.

[0124] In some embodiments, the ejection zone is away from the septum by at most about 0.2 mm, about 0.4 mm, about 0.6 mm, about 0.8 mm, about 1 mm, about 1.25 mm, about 1.5 mm, about 1.75 mm, about 2 mm, about 2.5 mm, or about 3 mm.

[0125] The dispensing element can be a dispensing element of a device for delivery of the delivery vehicle to a target region of the nasal cavity. The device can be inserted into the nasal cavity of the subject for delivery. The device can be inserted at an angle from a vertical line. In some embodiments, the vertical line is a line perpendicular to a horizontal line that is parallel to the inferior aspect of the olfactory cleft, as depicted as line 621 of FIG. 4J.

[0126] In some embodiments, the device is inserted into the nasal cavity of the subject at an angle of between about 30 degrees to about 40 degrees from a vertical line, the vertical line beingMintz Ref.: 062706-519001 WO perpendicular to a horizontal line that is parallel to an inferior aspect of the olfactory cleft of the subject. In some embodiments, the angle is about 30 degrees from the vertical line. In some embodiments, the angle is about 31 degrees from the vertical line. In some embodiments, the angle is about 32 degrees from the vertical line. In some embodiments, the angle is about 33 degrees from the vertical line. In some embodiments, the angle is about 34 degrees from the vertical line. In some embodiments, the angle is about 35 degrees from the vertical line. In some embodiments, the angle is about 36 degrees from the vertical line. In some embodiments, the angle is about 37 degrees from the vertical line. In some embodiments, the angle is about 38 degrees from the vertical line. In some embodiments, the angle is about 39 degrees from the vertical line. In some embodiments, the angle is about 40 degrees from the vertical line.

[0127] In some embodiments, the formulation has a viscosity of about 1 cP. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.0 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.1 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.2 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.3 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.4 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.5 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.6 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.7 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.8 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 1.9 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 2.0 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 2.1 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 2.2 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 2.3 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 2.4 m / s. In some embodiments, deliveringMintz Ref.: 062706-519001 WO the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 2.5 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation n from the ejection zone at a velocity of about 2.6 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 2.7 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 2.8 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 2.9 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of between about 1.1 m / s and about 1.9 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of between about 1.0 m / s and about 2.2 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of between about 1.0 m / s and about 2.8 m / s.

[0128] In some embodiments, the formulation has a viscosity of about 50 cP. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 4 m / s. In some embodiments, the formulation has a viscosity of about 50 cP. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 4.7 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 5 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 6 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 7 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 8 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 9 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 10 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 11 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 12 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 13 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 14 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 15 m / s. In some embodiments, delivering the formulation comprises ejecting theMintz Ref.: 062706-519001 WO formulation from the ejection zone at a velocity of about 16 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 17 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 18 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 18.6 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of about 19 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of between about 4.7 m / s and about 18.6 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of between about 13.9 m / s and about 15.8 m / s. In some embodiments, delivering the formulation comprises ejecting the formulation from the ejection zone at a velocity of between about 11.0 m / s and about 15.8 m / s.

[0129] As demonstrated herein, where the target region comprises an olfactory region, the formulation delivered by the method disclosed herein can remain at the olfactory cleft of the subject for a period of time longer than when the formulation is delivered by a nasal spray.

[0130] In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 10 seconds. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 30 seconds. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 1 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 2 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 3 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 4 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 5 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 6 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 7 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 8 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 9 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 10 min. In some embodiments, theMintz Ref.: 062706-519001 WO formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 11 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 12 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 13 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 14 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for at least about 15 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for longer than 15 min. In some embodiments, the formulation delivered to the olfactory cleft of the subject remains at the olfactory cleft for a period of time longer than if the formulation was delivered by nasal spray.

[0131] Delivery vehicles

[0132] Any suitable delivery vehicles may be used with the methods disclosed herein.

[0133] The term “delivery vehicle” is used broadly herein and encompasses any vehicle that can be used with a molecule for its delivery to a subject. The delivery vehicle can be nano-scale or microscale. Nano-scale delivery vehicles can include various nanoparticles, including but not limited to lipid nanoparticles, polymeric nanoparticles, and inorganic-based nanoparticles. Biological delivery vehicles, such as viral particles and exosomes, can also be used with the method disclosed herein. The biological delivery vehicles may be naturally occurring, or they may be modified. The delivery vehicle can be used to deliver various types of molecules, such as nucleic acids, peptides, proteins, small molecules, and combinations thereof.

[0134] In some embodiments, the delivery vehicle comprises a nucleic acid delivery vehicle. A nucleic acid delivery vehicle refers to a vehicle for delivering a nucleic acid molecule. Vehicles for delivering nucleic acid molecules include, for example, lipid nanoparticles, exosomes, and viral vectors. In some embodiments, the nucleic acid delivery vehicle comprises a lipid nanoparticle. In some embodiments, the nucleic acid delivery vehicle comprises an exosome. In some embodiments, the nucleic acid delivery vehicle comprises a viral vector. Examples of viral vectors include adenoviral vectors, adeno-associated viral (AAV) vectors, lentiviral vectors, and retroviral vectors. Different subtypes of the viral vectors can be used. For example, an AAV vector can be, but not limited to, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13 or any variants thereof.

[0135] In some embodiments, the delivery vehicle comprises a viral vector. In some embodiments, the viral vector is an adeno-associated viral (AAV) vector.Mintz Ref.: 062706-519001 WO

[0136] In some embodiments, the extracellular vesicle comprises an exosome.

[0137] In some embodiments, the exosome comprises a cargo. In some embodiments, the cargo is encapsulated by the exosome. In some embodiments, the cargo comprises a nucleic acid molecule, a small molecule, a protein, a peptide, or any combination thereof.

[0138] In some embodiments, the delivery vehicle comprises a lipid nanoparticle.

[0139] In some embodiments, the lipid nanoparticle comprises a nucleic acid molecule.

[0140] The nucleic acid molecule can comprise an RNA. Non-limited examples of RNA include messenger RNA (mRNA), microRNA (miRNA), small interfering RNA (siRNA), small activating RNA, self-amplifying RNA (saRNA), short hairpin RNA (shRNA), circular RNA, RNA decoy, silencing RNA, etc.

[0141] The RNA can be modified. For example, an mRNA may be capped at the 5’ end by, for example, a 7-methylguanosine cap analog.

[0142] In some embodiments, the lipid nanoparticle composition comprises an RNA. In some embodiments, the lipid nanoparticle composition comprises a vaccine. In some embodiments, the lipid nanoparticle composition comprises an RNA vaccine.

[0143] In some embodiments, the RNA vaccine comprises a mRNA vaccine. In some embodiments, the RNA vaccine comprises a saRNA vaccine.

[0144] In some embodiments, the lipid nanoparticle comprises a cargo. In some embodiments, the cargo is encapsulated by the lipid nanoparticle. In some embodiments, the cargo comprises a nucleic acid molecule, a small molecule, a protein, a peptide, or any combination thereof.

[0145] The lipid nanoparticle can be conjugated. For example, the lipid nanoparticle can be conjugated to an antigen-binding fragment, or an antibody or fragment thereof. The lipid nanoparticle can be conjugated to a drug.

[0146] The method disclosed herein can minimize shear forces exposed to the delivery vehicle.

[0147] In some embodiments, the formulation is delivered without exposing the delivery vehicle to shear forces sufficient to damage a significant portion of the delivery vehicle, without aerosolizing the formulation, or by passing the formulation through a shear disintegrating tip.

[0148] It is demonstrated herein that using the disclosed method to deliver lipid nanoparticles can result in more consistent particle size, with a diameter of less than about 100 nm and a polydispersity index (PDI) of less than about 0.2, when compared to lipid nanoparticles delivered with a nebulizer.Mintz Ref.: 062706-519001 WO

[0149] In some embodiments, the delivered delivery vehicle comprises a particle size of below about 150 nm, below about 140 nm, below about 130 nm, below about 120 nm, below about 110 nm, or below about 100 nm.

[0150] In some embodiments, the delivered delivery vehicle comprises a particle size that is within about 10%, within about 20%, within about 30%, within about 40%, or within about 50% of a particle size of a delivery vehicle prior to delivery.

[0151] In some embodiments, the delivered delivery vehicle comprises a poly dispersity index of less than about 0.2, less than about 0.19, less than about 0.18, less than about 0.17, less than about 0.16, less than about 0.15, less than about 0.14, less than about 0.13, less than about 0.12, less than about 0.11 , or less than about 0.1.

[0152] When formulating a delivery vehicle and cargo composition, a percentage of the cargo can be encapsulated. The higher the percentage of the cargo being encapsulated, the higher the encapsulation efficiency. As described herein, delivering the delivery vehicle composition with a delivery device can subject the delivery vehicles to shear forces, which can result in a reduced percentage of the cargo being encapsulated by the delivered delivery vehicles. For example, the integrity of the delivery vehicle may be compromised during delivery, resulting in a reduction in encapsulation efficiency. As demonstrated herein, the disclosed method herein can result in an encapsulation efficiency that is higher than an encapsulation efficiency when a nebulizer device is used for delivery.

[0153] In some embodiments, the method results in an encapsulation efficiency of the cargo by the delivery vehicle of at least about 2-fold- at least about 3-fold, at least about 4-fold, or at least about 5-fold an encapsulation efficiency resulting from a spray method when the same cargo and delivery vehicle are used.

[0154] In some embodiments, the method results in an encapsulation efficiency of an RNA cargo by a nanoparticle of at least about 2-fold- at least about 3-fold, at least about 4-fold, or at least about 5-fold an encapsulation efficiency the RNA cargo by the nanoparticle resulting from a spray method.

[0155] As used herein, the term “encapsulation efficiency” refers to the percentage of a cargo that is being encapsulated by the delivery vehicle. It can be calculated using the formula:Mass of encapsulated carqo— — - : -J 1- - — * - 11 / 0~\ / 0~\f%\ / Total mass of cargo usedMintz Ref.: 062706-519001 WO

[0156] As demonstrated herein, using the method disclosed herein, a higher total expression level of an mRNA encapsulated by LNP can be achieved, as compared to when the same amount of mRNA encapsulated by LNP is delivered using a nebulizer.

[0157] In some embodiments, the delivery vehicle encapsulates an RNA, wherein after a dose of the formulation is delivered, an expression level of the delivered RNA in a cell of the subject is at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 110%, at least about 110%, at least about 120%, at least about 130%, at least about 140%, at least about 150%, at least about 160%, at least about 170%, at least about 180%, at least about 190%, or at least about 200% higher than an expression level from a same dose of the formulation delivered by a spray method.

[0158] In some embodiments, the delivery vehicle is a lipid nanoparticle encapsulating an RNA, wherein after a dose of the formulation is delivered, an expression level of the delivered RNA in a cell of the subject is at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, or at least about 3-fold an expression level from a same dose of the formulation delivered by a spray method.

[0159] As used herein, the term “expression level of the delivered RNA”, when referring to an RNA encapsulated in the delivery vehicle delivered by the method disclosed herein, or by a comparison method (e.g. a non-targeted method, a non-liquid jet method, a spray method, etc), means the expression from all or substantially all RNA molecules delivered that is measurable. A higher expression level of the delivered RNA may be achieved in various ways. For example, a larger portion of the formulation and hence a higher percentage of the delivery vehicles encapsulating the RNA, may be taken up by cells. Alternatively, or additionally, a higher percentage of the delivery vehicles encapsulating the RNA may be intact upon delivery and hence a higher percentage of the RNA molecules are capable of being expressed.

[0160] In some embodiments, the delivery vehicle comprises a vaccine. In some embodiments, administering of the formulation results in an increased antigenicity in the subject compared to where the formulation is delivered to the subject as a spray at about the same dose.

[0161] In some embodiments, the formulation is delivered by a device, the device comprising: (a) a housing comprising an insertable portion comprising a distal end, and a proximal end; and (b) a subject-engaging portion which engages a columella region of the subject to seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject, wherein the device is configured to deliver the formulation as a laminar jet.Mintz Ref.: 062706-519001 WO

[0162] In some embodiments, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% of the formulation is delivered to the targeted region of the nasal cavity of the subject.

[0163] In some embodiments, the ejection zone is: (a) 0mm to 30mm superior to a horizontal line that intersects the anterior aspect of the internal nasal valve, and (b) 0mm to 20mm anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule.

[0164] In some embodiments, the ejection zone is further: (i) 0mm to 40mm inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft; (ii) 0mm to 20mm posterior to the internal nasal dorsum; (iii) 10mm to 50mm superior to a horizontal line that intersects the inferior aspect of the columella; (iv) 0mm to 30mm superior to a horizontal line that intersects the superior aspect of the inferior turbinate; (v) 0mm to 3mm from the septum, or; (vi) any combination thereof.II. Devices

[0165] In another aspect, provided herein is a device for intranasal delivery of a delivery vehicle to a targeted region of a nasal cavity of a subject, the device comprising: (a) a housing comprising an insertable portion comprising a distal end, and a proximal end; and (b) a subject-engaging portion which engages a columella region of the subject to seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject, wherein the device is configured to deliver a formulation comprising the delivery vehicle to the targeted region of the subject. In another aspect, provided herein is a device for delivery of a delivery vehicle to a target region of a nasal cavity of a subject. The device can comprise a housing comprising an insertable portion comprising a distal end and a proximal end. The device can comprise a subject-engaging portion which engages a columella region of the subject. The subject-engaging portion can seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject. The device can be configured to deliver a formulation comprising the delivery vehicle to the target region of the subject.

[0166] In some embodiments, the device dispenses the formulation as a laminar jet.

[0167] In some embodiments, the device comprises a compliant dispensing tip comprising a compliant and flexible soft nib.

[0168] In some embodiments, the dispensing tip has a diameter of between about 1.2 mm and 1.4 mm. In some embodiments, the dispensing tip has a diameter of about 1.3 mm. In some embodiments, the dispensing tip has a diameter of about 1.35 mm.Mintz Ref.: 062706-519001 WO

[0169] In some embodiments, the formulation has a viscosity of between about 0.5 cP and about 100 cP. In some embodiments, the device is configured to deliver the formulation at a velocity of between about 1 m / s and about 30 m / s.

[0170] In some embodiments, application of pressure by the subject-engaging portion to the columella region of the subject enables and / or causes delivery of the formulation to the subject from the insertable portion.

[0171] In some embodiments, the insertable portion comprises a dispensing element for delivery of the formulation to the target region of the subject.

[0172] The housing can comprise two insertable portions. In some embodiments, the two insertable portions comprise at least one dispensing element, each insertable portion for insertion into a nasal channel of the subject, wherein, upon insertion of at least one insertable portion into a nasal channel of the subject, the at least one insertable portion engages tissue within the nasal channel to open or expand an internal nasal valve of the subject thereby positioning the at least one dispensing element for delivery of the formulation to the subject. In some embodiments, the subject engaging portion comprises a trigger, wherein upon application of pressure to the subject engaging portion, the trigger permits actuation of the device to deliver the formulation to the subject through the dispensing element.

[0173] The method and device disclosed herein can be used for intranasal delivery. In some embodiments, the intranasal delivery comprises targeted delivery to a region in the nasal cavity. For example, the targeted region can be an olfactory region, a turbinate region (e.g. a middle turbinate region, a superior turbinate region, an inferior turbinate region), a nasal pharynx region, a nasal associated lymphatic tissue, etc. In some embodiments, the targeted region is an olfactory region. In some embodiments, the targeted region is a turbinate region. In some embodiments, the turbinate region is a middle turbinate region. In some embodiments, the targeted region is a nasal associated lymphatic tissue. In some embodiments, the targeted region is a nasopharynx region.

[0174] The device can be used with the methods disclosed herein.III. Definitions

[0175] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0176] The term “about” or “approximately” when used in reference to a particular recited value, means the value may vary from the recited value by no more than 10%, 5%, 2% or 1%. Where aMintz Ref.: 062706-519001 WO particular value is recited, it can be understood that the value is modified by the term “about” or “approximately”, unless indicated otherwise.

[0177] The term “comprise”, “comprising”, or the like means additional elements or components other than those recited may be present. Other terms such as “include”, “contain”, “have” and the like have similar meaning.

[0178] The term “consist of’, “consisting of’ or the like means no additional component is present.

[0179] As used herein, the singular forms “a,” “an,” and “the” include plural references, unless indicated otherwise. For example, a reference to “a molecule” can be a reference to more than one molecule.

[0180] The term “or” is used to mean “and / or”, unless it is indicated explicitly to refer to alternatives only. It should also be noted that the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise. These terms can convey that any combination is specifically contemplated. Solely for illustrative purposes, the expression “A, B, and / or C” can mean A individually; B individually; C individually; A and B; B and C; A and C; and A, B, and C.

[0181] As used herein, a “therapeutically effective amount” means an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that elicits a desired biological response. In some embodiments, a therapeutically effective amount of a substance is an amount that is sufficient, when administered as part of a dosing regimen to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat, diagnose, prevent, and / or delay the onset of the disease, disorder, and / or condition. As will be appreciated by those of ordinary skill in this art, the effective amount of a substance may vary depending on such factors as the desired biological endpoint, the substance to be delivered, the target cell or tissue, etc. For example, the effective amount of a provided compound in a formulation to treat a disease, disorder, and / or condition is the amount that alleviates, ameliorates, relieves, inhibits, prevents, delays onset of, reduces severity of and / or reduces incidence of one or more symptoms or features of the disease, disorder, and / or condition. In some embodiments, a “therapeutically effective amount” is at least a minimal amount of a provided compound, or composition containing a provided compound, which is sufficient for treating one or more symptoms of a disease or disorder.

[0182] The term “lipid nanoparticle” as used herein refers to a nano-scale particle comprising one or more lipids. The particle may comprise a lipid core or an aqueous core (e.g. a liposome).

[0183] The term “nucleic acid” or “nucleic acid molecule” refers to covalently linked sequence of nucleotides, such as ribonucleotides and deoxyribonucleotides, and encompasses analogs andMintz Ref.: 062706-519001 WO derivatives of natural nucleotides and modifications. A nucleic acid molecule may be formed by a single type of nucleotides (e.g. DNA or RNA), or it may be a hybrid of different nucleotides, for example, a DNA / RNA hybrid. A nucleic acid molecule may be single- stranded, double-stranded, or partially double-stranded. For example, a nucleic acid molecule may have a stem-loop structure, where the stem portion is double-stranded and the loop portion is single-stranded. A nucleic acid may be in different conformations, such as linear or circular.

[0184] The term “vector” means a vehicle capable of transferring exogenous genetic material into a cell. Examples of vectors include but are not limited to plasmids, cosmids, bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), and bacteriophages.

[0185] The term “viral vector” means a vector with at least one element of viral origin.

[0186] The “columella” is the firm tissue bridge that separates the nostrils at the base of the nose. The “columella” is the most anteroinferior portion of the nasal septum. The term “columella” or “columella region” is the subnasale, or an anterior nasale spine, or a combination thereof. The columella region may comprise a subnasale, or a combination thereof. The columella shape may be defined by an anterior nasal spine located posteriorly to the columella e.g., 1 cm.

[0187] The term “composition” or “compound” or “therapeutic” or “sampling compound” or “sampling fluid” is therapeutics, medicaments, drugs, small and large molecules, medicaments in liquid, powder, or gas form, or a combination thereof having a low, intermediate, or high viscosity.

[0188] The “introduction pathway” is, in sequence, the vestibule, the anterior aspect of the internal nasal valve, and the anterior aspect of the respiratory region - anterior of the turbinates.

[0189] The “internal nasal valve” (INV) is a space bounded medially by the dorsal septum 24 (or just septum), laterally by the caudal portion of the upper lateral cartilage, and inferiorly by the head of the inferior turbinate. This is the narrowest part of the nasal airway, located just beyond the nostril. It's formed by the edge of the nasal septum, the upper lateral cartilage, and the floor of the nose. The internal nasal valve plays a critical role in regulating airflow through the nose.

[0190] The term “nasal cavity” refers to the large, air-filled space behind the nose, where air passes on its way to the throat during inhalation and includes two nasal channels, each comprised of a vestibule, respiratory region and olfactory cleft, and a nasopharynx.

[0191] The term “trigger” refers to the part of the device that actuates the ejection mechanism, which in turn (through a variety of possible mechanism designs), delivers a compound from the device.

[0192] The term “turbinates” refers to superior turbinate, middle turbinate, or inferior turbinate, or a combination thereof.Mintz Ref.: 062706-519001 WO

[0193] Nasal Septum: This is the thin wall of bone and cartilage that separates the right and left nostrils. It forms the medial (towards the middle) boundary of the region of interest.

[0194] Lateral Nasal Wall: This is the side wall of the nasal cavity, which is opposite to the nasal septum. It's a complex structure that includes the turbinates (long, curled bones that protrude into the nasal cavity) and the meatuses (grooves or channels between the turbinates). The lateral nasal wall forms the lateral (towards the side) boundary of the region of interest.

[0195] Middle and Superior Meatuses: These are the spaces within the nasal cavity located between the turbinates. The middle meatus is located beneath the middle turbinate and above the inferior turbinate, and the superior meatus is located beneath the superior turbinate. The region of interest encompasses parts of these spaces.

[0196] Nostrils (External Nares): These are the two openings of the nose where air enters.

[0197] Nasal Vestibule: The nasal vestibule is the most anterior part of the nasal cavity, just inside the nostrils. It's the area of the nose that protrudes outside the face predominantly. This area is lined with skin and contains hair follicles, and it acts as the initial filtering and warming area for inhaled air before it moves deeper into the nasal cavity. The nasal vestibule extends posteriorly to the nasal valve, which is the narrowest part of the nasal airway and located just beyond the nostril.

[0198] Nasal Septum: This is a thin wall made of bone and cartilage that separates the left and right sides of the nasal cavity.

[0199] Turbinates (Nasal Conchae): These are three pairs of bony projections (inferior, middle, and superior) covered in mucous membrane that protrude into the nasal cavity from the lateral walls. They increase the surface area of the nasal cavity, aiding in the warming, humidification, and filtration of inhaled air.

[0200] Meatuses: These are the spaces located between the turbinates. Each turbinate has a corresponding meatus underneath it (i.e., inferior, middle, and superior meatus).

[0201] Olfactory Region: This is a small area located at the top of the nasal cavity, where the sense of smell is located.

[0202] The above disclosure generally describes the present application. A more complete understanding can be obtained by reference to the following specific examples. These examples are described solely for the purpose of illustration and are not intended to limit the scope of the application. Changes in form and substitution of equivalents are contemplated as circumstances might suggest or render expedient. Although specific terms have been employed herein, such terms are intended in a descriptive sense and not for purposes of limitation.Mintz Ref.: 062706-519001 WOEXAMPLESExample 1: Laminar Fluid Ejection Method

[0203] The primary goal of the study was to develop a method of ejecting fluid through a cannula for deposition at the olfactory cleft.

[0204] Aperture testing was conducted to evaluate the coherence of fluid post-ejection and to visually assess the laminar qualities of the ejected fluid. A device comprising the cannula was secured in place for the aperture testing. Fluids at two different viscosities (1 cP and 50 cP) were tested, each at a range of ejection velocities (2 m / s - 27.7 m / s). Device settings for a selection of velocities within this range were determined experimentally prior to conducting aperture testing. Velocity was determined using high-speed camera-footage of ejected fluid traveling a fixed distance from the tip of the canula. A small aperture (5.64 mm in diameter) was placed 25 mm away from the tip of the canula. The canula was supported by magnetic supports to reduce wobbling. A catching tray was placed on the opposite side of the aperture. The percentage mass transferred from the syringe to the catching tray was calculated for each test-run (Mass in catching tray / Mass Ejected from syringe * 100). Tests of each combination of viscosity and speed were repeated three times. The mean percent mass transferred for each condition is presented in Table 1. (Note that the ejected mass is smaller than the total fill mass, as it does not include the residual left in the device. The residual left in the device did not meaningfully differ across conditions: 13.6% for 1 cP, 10.4% for 50 cP).Table 1. Aperture TestingPercent mass transferred from the device through a small 5.64 mm aperture and into a catching tray for low / high fluid viscosities and velocities.

[0205] In general, higher speeds were associated with reduced mass transfer, possibly due to increased cannula movement and splattering off the aperture edge. Qualitatively, it was observed that the device ejected fluid in a cohesive stream with a narrow diameter, especially compared to the plume emitted from a traditional spray device.Mintz Ref.: 062706-519001 WO

[0206] Next, a series of in-vitro deposition tests was conducted to determine the range of velocities that would optimally deposit fluid in the olfactory cleft of the human nasal passages. To test this, the mechanism was inserted into a transparent 3D-printed model of the nasal cavity. Optimal delivery was considered to be in the target zone, which represents the approximate location of the olfactory cleft in the model. If ejected too slowly, the fluid would deposit anterior to the front boundary of the target zone; too quickly and it would deposit beyond the back boundary. Two fluid viscosities (1 cP and 50 cP) were tested at each of three different insertion angles (30, 35, 40 degrees from the vertical). The cannula’s insertion depth was held constant at 37.5 mm. The minimum velocity necessary to reach the Front Boundary (vf), the olfactory cleft (vopt), and the back boundary (vb), for each of the viscosities and deposition angles are presented in Table 2.Table 2. In-vitro Deposition TestingTable presents optimal velocities (Vopt) for fluids of low / high viscosity across three device angles. The Vf and Vb columns show the velocity limits at which the deposition was in front of or behind the target area, respectively.

[0207] For velocities within the optimal range, residence times were observed to exceed 10 minutes. Notably, angles of 35-40 degrees permitted a wider range of velocities, but using an angle of 30- degrees did not dramatically shift the optimal velocity. This suggested the device is adaptable to a wide range of angles. Optimal velocities (+ / -20%) were found to be 1.6 m / s for 1 cP fluid and 14.0 m / s for 50 cP fluid. These optimal velocities may be associated with laminar flow.

[0208] A Reynold’s number (Re= p* V*L / p, where p is the density of the fluid, V is the velocity of the flow, L is the diameter of the passage, and p is the dynamic viscosity of the fluid) can be computed to quantify whether a fluid exhibits laminar or turbulent flow. The Reynold’s number for water at a velocity of 1.6m / s and viscosity of IcP is approximately 2,160 with a 1.35 mm diameter (density of water is approximately 1000 kg / m3). Similarly for a fluid of 50cP (with the same density as water), the Reynold’s number at 15m / s is approximately equal to 405 with a 1.35 mm diameter. Both Reynold’s numbers fall below 2,300 which is the commonly accepted threshold for laminar flow.Mintz Ref.: 062706-519001 WOExample 2: In-Vivo Evaluation of Laminar Fluid Ejection Method

[0209] The primary goal of these studies is to visualize delivery to the olfactory cleft (first with technetium-99, and then with methylene blue). The device was tuned to eject the fluid at a specific velocity. This velocity was chosen based on the viscosity of the fluid and the studies in Example 1, to maximize the extent of laminar flow.

[0210] Visualizing Olfactory Cleft Delivery with Technetium-99

[0211] Participants'. Nine healthy participants (ages 19+) with no history of abnormal nasal or sinus symptoms or contraindications for nasal cannulation, Magnetic Resonance Imaging (MRI), or SinglePhoton Emission Computed Tomography (SPECT). Participants completed one study visit undergoing magnetic resonance imaging (MRI) and single-photon emission computed tomography (SPECT) to visualize the deposition of a radiolabeled tracer (technetium-99) in their nasal passages. In total, nine participants were enrolled in the study, and eight completed all study procedures (Participant 1.02 withdrew before completing all imaging procedures). All study procedures were approved by the Horizon Health Network’s Human Research Protection Program. Prior to participating in the study, all participants were assessed by a licensed otolaryngologist physician to confirm their eligibility. Study Design: On Day 1 of the study, anatomical (T1 -weighted) magnetic resonance images were collected from participants at the Moncton MRI clinic (Moncton, NB, Canada) using a Siemens Skyra VD13 3T scanner. MRI data provided anatomical information regarding soft tissue structure in the nasal cavity and the location of each participant’s olfactory region.

[0212] On Day 2 of the study, a saline solution including the technetium-99 radiotracer (mean dose 4.6 mCi) was delivered to participants using the Laminar Fluid Ejection method described above. Day 2 study procedures were conducted at the Nuclear Imaging Department of Saint John Regional Hospital (Saint John, NB, Canada). At the time of delivery and for five minutes afterwards, 2D SPECT Flow images were acquired every three seconds using a gamma camera. Flow images were acquired in a single sagittal plane (128x128 with voxels 4.8mm diameter).

[0213] Image Analysis: Image analysis was completed in Vivoquant (4.0). SPECT Flow data was resampled to match MRI resolution (320 x 320 with voxels 1.918mm isotropic) and manually registered to T1 images in x and y space with minimal rotation in the z-plane. This procedure enabled landmarking to nasal passage characteristics minimally visible in the SPECT data alone. Flow data was visually inspected to determine (1) the point in the time series when the bolus was dispensed, (2) the time of cannula removal, (3) the median time point in the series where the bolus initially lodged between points 2 and 4, (4) the time point at which the bolus began additional migration, and (5) theMintz Ref.: 062706-519001 WO time point of the final image acquired during the five-minute SPECT series. All subjects’ SPECT Flow images are presented in FIG. 8.

[0214] Results: In seven out of eight participants imaging results demonstrated delivery of the radiotracer to the cribriform area. The radiotracer was detected in the cribriform area without any movement for at least 1.5 minutes in six of those seven participants. Additionally, clearance of the radiotracer was minor for the duration of the study for six out of those seven cases. In the two cases where sub-optimal outcomes were reached, it was the result of (i) bolus delivery anterior and superior to the cribriform (Participant 1.07), and (ii) bolus that cleared quickly after reaching the target, possibly due to the bolus remaining well intact as it directly impacted the cribriform (Participant 1.05). See Table 3 for a summary of the results in each participant.Table 3. Visualizing Olfactory Cleft Delivery with Technetium-99

[0215] Visualizing Olfactory Cleft Delivery with Methylene Blue

[0216] Participants'. Five healthy participants (ages 19+) with no history of sinonasal symptoms and no evidence of nasal inflammation were recruited for this study. Prior to recruitment of participants study procedures were approved by Providence Health Care research ethics board, (British Columbia, Canada) and conducted under Health Canada Investigational Testing Authorization #314993. All participants were assessed by a licensed otolaryngologist physician to confirm their eligibility. Participants completed two study visits during which a licensed otolaryngologist physician administered 0.1 mF of the visual dye tracer, methylene blue.

[0217] Study Design: During the first visit, the methylene blue was administered using the Eaminar Fluid Ejection method (velocity = 4.5 m / s). During the second visit, methylene blue was administered using a conventional nasal spray device (Pharma systems Item #10272, UPC:063636802714). The Pharma systems spray device was used in these experiments because it was the device normally usedMintz Ref.: 062706-519001 WO by the compounding pharmacy for intranasal delivery. It was the device patients and clinicians used at this clinic.

[0218] Following delivery, the physician used a nasal endoscope to image the deposition of methylene blue to the olfactory cleft at up to five time points (1, 5, 8, 12, and 15 minutes following delivery). From each image, the physician judged whether at least 50% of the methylene blue fluid was delivered to the olfactory cleft. If <50% of the methylene blue was judged to have deposited in the olfactory cleft, imaging was stopped and that session’s trial was completed.

[0219] Results: Four of the five participants demonstrated successful delivery of >50% of the methylene blue to the olfactory cleft using the Laminar Fluid Ejection method. In each of the five participants, the conventional nasal spray failed to deliver at least 50% of the dye to the olfactory cleft (Table 4).Table 4. Visualization of Delivery to the Olfactory Clef Using Methylene BlueResults from in-human tests of the Laminar Fluid Ejection (LFE) method using the dye methylene blue.Example 3 Characterization of lipid nanoparticles delivered by a low-shear device vs a high- shear device

[0220] The objective of this study is to characterize lipid nanoparticle (LNP) formulations that have been ejected from a low-shear device compared to a high-shear device (nebulizer). The characterization aims to assess the impact of device-driven administration (low-shear and high-shear devices) on qualities such as particle size, polydispersity, and encapsulation efficiency.

[0221] Method

[0222] Luciferase mRNA and luciferase saRNA were formulated as lipid nanoparticles. A lipid mix was prepared in the ethanol phase, consisting of ALC-0315:DOPE:Cholesterol:DMG-PEG-2000 at molar ratios of 45:17.5:36.25:1.25, respectively. Using a T-Junction mixer, the lipid and aqueousMintz Ref.: 062706-519001 WO phases were rapidly mixed at a ratio of 10, maintaining a total flowrate of 20 mL / min with a 1:3 flowrate ratio (lipids to RNA). Each formulation was subject to the low-shear device and the high- shear device (nebulizer). As a control, the formulations were not subject to either of the devices.

[0223] FIG. 15 shows the setup of a proprietary low-shear device (RSH device). The RSH device dispenses a formulation or pushes a plunger arm at a rate of 2 m / s. The RSH device was mounted onto a test sled. A 15 mL tube used to collect the dispensed LNP formulation was inserted onto the test sled from the other side. 200uL of the LNP formulation was loaded into a syringe. A cannula was attached to the luer fitting of the syringe. The cannula, attached to the syringe, was inserted through the hole in the cap of the 15 mL tube. After inserting the cannula inside the 15 mL tube, it was positioned 8 mm away from the side boundary of the tube and tilted at a 5-degree angle with respect to that boundary. This positioning aimed to simulate the nasal anatomy. RSH device’s push mechanism was utilized to push the syringe plunger, dispensing the LNP formulation into the 15 mL tube. Using the RSH device caused the formulation to strike the wall of the 15 mL tube. This replicated the interaction between the administered formulation and the nasal anatomy.

[0224] FIG. 16 shows the setup of a spray device (MAD device). In this setup, a support compatible with the MAD device was used. The RSH device was mounted onto a test sled. A 15 mL tube for collecting the dispensed LNP formulation was inserted into the test sled. 200uL of the LNP formulation was loaded into a syringe. The MAD device was attached to the luer fitting of the syringe. The MAD device was positioned to connect to the syringe on the support structure. The RSH device’s push mechanism was used to push the syringe plunger, dispensing the LNP formulation into the 15mL tube. The formulation was then dispensed through the MAD device inside the 15mL tube.

[0225] Particle size, polydispersity index, encapsulation efficiency, and zeta potential were measured. The processed formulations were used to transfect HEK293T cells and expression of luciferase was measured at the 24-hour time point.

[0226] The encapsulation efficiency within LNPs was measured using the Quant-iT Ribogreen RNA assay kit (Invitrogen, Thermo FisherScientific) as previously described [Ly et al, 2022, Molecular Pharmaceuticals 19:1892-1905]. Lipid nanoparticles were analyzed for particle size, polydispersity index (PDI), and zeta potential by dynamic light scattering (DLS) using the Zetasizer Nano (Malvern Instruments). For these measurements, the following parameters were applied: a material refractive index of 1.4, an absorbance of 0.01, a dispersant viscosity of 0.882 cP, a refractive index of 1.33, and a dielectric constant of 79. The nanoparticles were diluted 10 folds before analysis, and three measurements were collected for each sample.Mintz Ref.: 062706-519001 WO

[0227] Results

[0228] Processing of the formulation with the low-shear device resulted in particle size of < 100 nm and more consistent particle size. Processing with the nebulizer resulted in increased particle size (FIGs. 9A-9B).

[0229] Processing of the formulation with the low-shear device gave better distribution compared to the nebulizer: processing with the low-shear device resulted in PDI of <0.2 nm. Processing with the nebulizer resulted in increased PDI (FIGs. 10A-10B).

[0230] Encapsulation efficiency of greater than 80% was achieved by the low-shear device. Processing with the nebulizer resulted in reduced encapsulation efficiency (FIGs. 11A-11B and 12A- 12B).

[0231] Both devices were able to achieve a zeta potential of ± 4 mV (FIGs. 13A-13B).

[0232] LNP formulations were used to transfect HEK293T cells. Luciferase assay was performed at the 24-hour time point. FIGs. 14A-14D show that higher expression levels were achieved with the low-shear device compared with the high-shear device.Example 4- mRNA-LNP biodistribution

[0233] Background: The ability of a drug to reach its target can be affected by its route of administration. Standard routes of administration include intravenous (IV) and intranasal (IN). Intravenous injections can cause patient distress and require a trained phlebotomist to administer the drug. Intranasal sprays are often preferred by patients because of their ease of use, but suffer from imprecise dosing and delivery. The goal of this example was to compare nasal cannula delivery (OD) with the traditional IV and IN administration routes. In this study, luciferase mRNA formulated in lipid nanoparticles was administered to mice via different delivery methods. Expression of the mRNA was then monitored.

[0234] Methods

[0235] LNP -luciferase mRNA was prepared as in Example 3. Three different delivery methods were used: (1) Olfactory (OD): A 0.699 mm diameter cannula was used to simulate olfactory delivery; (2) Intranasal (IN): A p200 standard Pipette tip was placed into the nostril just up to the nasal valve to simulate intranasal spray delivery; (3) Intravenous (IV): Intravenous delivery via tail vein to serve as uptake control.

[0236] Mice were subjected to different treatments (Table 5) and then imaged with the IVIS (PerkinElmer) at 1, 3, 6, 8, 24, 48, and 72 hours post-dosing for whole-body bioluminescence imagingMintz Ref.: 062706-519001 WO(prone position and supine position). 15 minutes before imaging, mice were intraperitoneally injected with 150 mpk D-luciferin.Table 5. Treatments

[0237] Results

[0238] As shown in Figs. 17A-17B, OD delivery resulted in higher levels of bioluminescence for over 24 hours post-dosing compared to IN and IV delivery. As shown in Figs 18A-18G, bioluminescence is localized in the heads of the mice for both OD and IN delivery.

[0239] Enumerated EmbodimentsEnumerated Embodiment 1. A method of delivering a lipid nanoparticle to a subject in need thereof, the method comprising: delivering a formulation comprising the lipid nanoparticle to a targeted region of a nasal cavity of the subject, wherein the formulation is delivered as a liquid jet, wherein the delivered lipid nanoparticle comprises a particle size that is within about 50% of a particle size of the lipid nanoparticle prior to delivery.Enumerated Embodiment 2. A method of delivering a delivery vehicle to a subject in need thereof, the method comprising: delivering a formulation comprising the delivery vehicle to a targeted region of a nasal cavity of the subject, wherein the formulation is delivered as a liquid jet.Enumerated Embodiment 3. The method of any of the enumerated embodiments, wherein the liquid jet is a laminar jet.Enumerated Embodiment 4. The method of any of the enumerated embodiments, wherein the formulation is delivered as a flow having a Reynold’s number of 2300 or less.Enumerated Embodiment 5. The method of any of the enumerated embodiments, wherein the delivering the formulation as a laminar flow, a liquid jet, or a flow having a Reynold’sMintz Ref.: 062706-519001 WO number of 2300 or less: a) increases on target delivery of the composition to the target region, b) decreases off target delivery of the composition to the nasal cavity, or c) both, compared to delivering the formulation with a spray ejection profile.Enumerated Embodiment 6. The method of any of the enumerated embodiments, wherein the formulation is ejected from a dispensing element, wherein at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% of the formulation passes through a circular opening having a diameter of between about 5 mm and about 6 mm at a distance of about 25 mm from where the formulation is ejected from the dispensing element.Enumerated Embodiment 7. The method of any of the enumerated embodiments, wherein the formulation has a viscosity of between about 0.5 cP and about 100 cP.Enumerated Embodiment 8. The method of any of the enumerated embodiments, wherein the formulation has a viscosity of between about 0.5 cP and about 50 cP.Enumerated Embodiment 9. The method of any of the enumerated embodiments, wherein the formulation has a viscosity of between about 0.5 cP and about 10 cP.Enumerated Embodiment 10. The method of any one of claims 1-8, wherein the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 30 m / s.Enumerated Embodiment 11. The method of any one of claims 1-8, wherein the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 5 m / s.Enumerated Embodiment 12. The method of any of the enumerated embodiments, wherein the formulation is ejected from a dispensing element, wherein at least 75% of the formulation passes through a circular opening having a diameter of between about 5 mm and about 6 mm at a distance of about 25 mm from where the formulation is ejected from the dispensing element, wherein the formulation has a viscosity of between about 0.5 cP and about 50 cP, and wherein the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 30 m / s.Enumerated Embodiment 13. The method of any of the enumerated embodiments, wherein at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% of the formulation is delivered to the targeted region of the nasal cavity of the subject.Enumerated Embodiment 14. The method of any of the enumerated embodiments, wherein the delivery vehicle comprises a macromolecule delivery vehicle.Mintz Ref.: 062706-519001 WOEnumerated Embodiment 15. The method of any of the enumerated embodiments, wherein the delivery vehicle comprises an exosome.Enumerated Embodiment 16. The method of any of the enumerated embodiments, wherein the exosome comprises a cargo.Enumerated Embodiment 17. The method of any of the enumerated embodiments, wherein the cargo is encapsulated by the exosome.Enumerated Embodiment 18. The method of any of the enumerated embodiments, wherein the delivery vehicle comprises a nucleic acid delivery vehicle.Enumerated Embodiment 19. The method of any of the enumerated embodiments, wherein the nucleic acid delivery vehicle comprises a viral vector.Enumerated Embodiment 20. The method of any of the enumerated embodiments, wherein the viral vector comprises an adeno-associated vector (AVV).Enumerated Embodiment 21. The method of any of the enumerated embodiments, wherein the method results in an encapsulation efficiency of at least about 2-fold- at least about 3- fold, at least about 4-fold, or at least about 5-fold an encapsulation efficiency by a spray method.Enumerated Embodiment 22. The method of any of the enumerated embodiments, wherein the delivery vehicle comprises a lipid nanoparticle.Enumerated Embodiment 23. The method of any of the enumerated embodiments, wherein the delivered lipid nanoparticle comprises a particle size of less than about 100 nm.Enumerated Embodiment 24. The method of any of the enumerated embodiments, wherein the delivered lipid nanoparticle comprises a particle size that is within about 10%, within about 20%, within about 30%, within about 40%, or within about 50% of a particle size of a delivery vehicle prior to delivery.Enumerated Embodiment 25. The method of any of the enumerated embodiments, wherein the delivered lipid nanoparticle comprises a poly dispersity index (PDI) of less than about 0.2.Enumerated Embodiment 26. The method of any of the enumerated embodiments, wherein the lipid nanoparticle encapsulates an RNA, wherein after a dose of the formulation is delivered, an expression level of the delivered RNA in a cell of the subject is at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, or at least about 3-fold an expression level from a same dose of the formulation delivered by a spray method.Mintz Ref.: 062706-519001 WOEnumerated Embodiment 27. The method of any of the enumerated embodiments, wherein the lipid nanoparticle composition comprises a nucleic acid molecule.Enumerated Embodiment 28. The method of any of the enumerated embodiments, wherein the lipid nanoparticle composition comprises a vaccine.Enumerated Embodiment 29. The method of any of the enumerated embodiments, wherein the vaccine comprises an RNA vaccine.Enumerated Embodiment 30. The method of any of the enumerated embodiments, wherein the RNA vaccine comprises a mRNA vaccine.Enumerated Embodiment 31. The method of any of the enumerated embodiments, wherein the RNA vaccine comprises a saRNA vaccine.Enumerated Embodiment 32. The method of any of the enumerated embodiments, wherein administering of the formulation results in an increased antigenicity in the subject compared to where the formulation is delivered to the subject as a spray at about the same dose.Enumerated Embodiment 33. The method of any of the enumerated embodiments, wherein the lipid nanoparticle composition comprises a cargo.Enumerated Embodiment 34. The method of any of the enumerated embodiments, wherein the cargo is encapsulated by the lipid nanoparticle.Enumerated Embodiment 35. The method of any of the enumerated embodiments, wherein the cargo comprises a nucleic acid molecule, a small molecule, a protein, a peptide, or any combination thereof.Enumerated Embodiment 36. The method of any of the enumerated embodiments, wherein the targeted region is an olfactory region.Enumerated Embodiment 37. The method of any of the enumerated embodiments, wherein the targeted region is a turbinate region.Enumerated Embodiment 38. The method of any of the enumerated embodiments, wherein the turbinate region is a middle turbinate region.Enumerated Embodiment 39. The method of any of the enumerated embodiments, wherein the targeted region is a nasal associated lymphatic tissue.Enumerated Embodiment 40. The method of any of the enumerated embodiments, wherein the targeted region is a nasopharynx region.Enumerated Embodiment 41. The method of any of the enumerated embodiments, wherein the formulation is delivered by a device, the device comprising:Mintz Ref.: 062706-519001 WO(a) a housing comprising an insertable portion comprising a distal end, and a proximal end; and(b) a subject-engaging portion which engages a columella region of the subject to seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject, wherein the device is configured to deliver the formulation as a laminar jet.Enumerated Embodiment 42. The method of any of the enumerated embodiments, wherein the ejection zone is:(a) Omm to 30mm superior to a horizontal line that intersects the anterior aspect of the internal nasal valve, and(b) Omm to 20mm anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule.Enumerated Embodiment 43. The method of any of the enumerated embodiments, wherein the ejection zone is further: i) Omm to 40mm inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft, ii) Omm to 20mm posterior to the internal nasal dorsum, iii) 10mm to 50mm superior to a horizontal line that intersects the inferior aspect of the columella, iv) Omm to 30mm superior to a horizontal line that intersects the superior aspect of the inferior turbinate, v) Omm to 3 mm from the septum, or vi) any combination thereof.Enumerated Embodiment 44. A device for intranasal delivery of a delivery vehicle to a targeted region of a nasal cavity of a subject, the device comprising: a) a housing comprising an insertable portion comprising a distal end, and a proximal end; and b) a subject-engaging portion which engages a columella region of the subject to seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject;Mintz Ref.: 062706-519001 WO wherein the device is configured to deliver a formulation comprising the delivery vehicle to the targeted region of the subject.Enumerated Embodiment 45. The device of any of the proceeding claims, wherein the device dispenses the formulation as a laminar jet.Enumerated Embodiment 46. The device of any of the proceeding claims, wherein the device comprises a compliant dispensing tip comprising a compliant and flexible soft nib.Enumerated Embodiment 47. The device of any of the proceeding claims, wherein the dispensing tip has a diameter of between about 1.2 mm and 1.4 mm.Enumerated Embodiment 48. The device of any of the proceeding claims, wherein the dispensing tip has a diameter of about 1.3 mm.Enumerated Embodiment 49. The device of any of the proceeding claims, wherein the dispensing tip has a diameter of about 1.35 mm.Enumerated Embodiment 50. The device of any of the proceeding claims, wherein the formulation has a viscosity of between about 0.5 cP and about 100 cP.Enumerated Embodiment 51. The device of any of the proceeding claims, wherein the device is configured to deliver the formulation at a velocity of between about 1 m / s and about 30 m / s.Enumerated Embodiment 52. The device of any of the proceeding claims, wherein the device is configured to deliver the formulation at a velocity of between about 1 m / s and about 5 m / s.Enumerated Embodiment 53. The device of any of the proceeding claims, wherein application of pressure by the subject-engaging portion to the columella region of the subject enables and / or causes delivery of the formulation to the subject from the insertable portion.Enumerated Embodiment 54. The device of any of the proceeding claims, wherein the insertable portion comprises a dispensing element for delivery of the formulation to the targeted region of the subject.Enumerated Embodiment 55. The device of any of the enumerated embodiments, wherein the ejection zone is:(a) 0mm to 30mm superior to a horizontal line that intersects the anterior aspect of the internal nasal valve, and(b) 0mm to 20mm anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule.Mintz Ref.: 062706-519001 WOEnumerated Embodiment 56. The device of any of the enumerated embodiments, wherein the ejection zone is further: vii) Omm to 40mm inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft, viii) Omm to 20mm posterior to the internal nasal dorsum, ix) 10mm to 50mm superior to a horizontal line that intersects the inferior aspect of the columella, x) Omm to 30mm superior to a horizontal line that intersects the superior aspect of the inferior turbinate, xi) Omm to 3mm from the septum, or xii) any combination thereof.Enumerated Embodiment 57. The device of any of the enumerated embodiments, configured to deliver at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% of the formulation to the targeted region of the nasal cavity of the subject.Enumerated Embodiment 58. The device of any of the enumerated embodiments, wherein the delivery vehicle comprises a macromolecule delivery vehicle.Enumerated Embodiment 59. The device of any of the enumerated embodiments, wherein the delivery vehicle comprises an exosome.Enumerated Embodiment 60. The device of any of the enumerated embodiments, wherein the exosome comprises a cargo.Enumerated Embodiment 61. The device of any of the enumerated embodiments, wherein the cargo is encapsulated by the exosome.Enumerated Embodiment 62. The device of any of the enumerated embodiments, wherein the delivery vehicle comprises a nucleic acid delivery vehicle.Enumerated Embodiment 63. The device of any of the enumerated embodiments, wherein the nucleic acid delivery vehicle comprises a viral vector.Enumerated Embodiment 64. The device of any of the enumerated embodiments, wherein the viral vector comprises an adeno-associated vector (AVV).Enumerated Embodiment 65. The device of any of the enumerated embodiments, wherein the delivery vehicle comprises a lipid nanoparticle.Mintz Ref.: 062706-519001 WOEnumerated Embodiment 66. The device of any of the enumerated embodiments, configured to deliver a lipid nanoparticle comprising a particle size of less than about 100 nm.Enumerated Embodiment 67. The device of any of the enumerated embodiments, configured to deliver a lipid nanoparticle comprising a particle size that is within about 10%, within about 20%, within about 30%, within about 40%, or within about 50% of a particle size of the lipid nanoparticle prior to delivery.Enumerated Embodiment 68. The device of any of the enumerated embodiments, configured to deliver a lipid nanoparticle comprising a polydispersity index (PDI) of less than about 0.2.Enumerated Embodiment 69. The device of any of the enumerated embodiments, configured to deliver the delivery vehicle with an encapsulation efficiency at least about 2-fold- at least about 3-fold, at least about 4-fold, or at least about 5-fold an encapsulation efficiency by a spray device.Enumerated Embodiment 70. The device of any of the enumerated embodiments, configured such that when the delivery vehicle comprises a lipid nanoparticle encapsulating an RNA, after a dose of the formulation is delivered, an expression level of the delivered RNA in a cell of the subject is at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, or at least about 3-fold an expression level from a same dose of the formulation delivered by a spray device.Enumerated Embodiment 71. The device of any of the enumerated embodiments, wherein the lipid nanoparticle composition comprises a nucleic acid molecule.Enumerated Embodiment 72. The device of any of the enumerated embodiments, wherein the lipid nanoparticle composition comprises a vaccine.Enumerated Embodiment 73. The device of any of the enumerated embodiments, wherein the vaccine comprises an RNA vaccine.Enumerated Embodiment 74. The device of any of the enumerated embodiments, wherein the RNA vaccine comprises a mRNA vaccine.Enumerated Embodiment 75. The device of any of the enumerated embodiments, wherein the RNA vaccine comprises a saRNA vaccine.Enumerated Embodiment 76. The device of any of the enumerated embodiments, wherein the lipid nanoparticle composition comprises a cargo.Enumerated Embodiment 77. The device of any of the enumerated embodiments, wherein the cargo is encapsulated by the lipid nanoparticle.Mintz Ref.: 062706-519001 WOEnumerated Embodiment 78. The device of any of the enumerated embodiments, wherein the cargo comprises a nucleic acid molecule, a small molecule, a protein, a peptide, or any combination thereof.Enumerated Embodiment 79. The device of any of the enumerated embodiments, wherein the targeted region is an olfactory region.Enumerated Embodiment 80. The device of any of the enumerated embodiments, wherein the targeted region is a turbinate region.Enumerated Embodiment 81. The device of any of the enumerated embodiments, wherein the turbinate region is a middle turbinate region.Enumerated Embodiment 82. The device of any of the enumerated embodiments, wherein the targeted region is a nasal associated lymphatic tissue.Enumerated Embodiment 83. The device of any of the enumerated embodiments, wherein the targeted region is a nasopharynx region.

[0240] While the present disclosure has been described with reference to examples, it is to be understood that the scope of the claims should not be limited by the embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole.

[0241] All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present disclosure is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.

[0242] The scope of the claims should not be limited by the embodiments and examples, but should be given the broadest interpretation consistent with the description as a whole.

Claims

Mintz Ref.: 062706-519001 WOCLAIMSWhat is claimed is:

1. A method of delivering a lipid nanoparticle to a subject in need thereof, the method comprising: delivering a formulation comprising the lipid nanoparticle to a targeted region of a nasal cavity of the subject, wherein the formulation is delivered as a liquid jet, wherein the delivered lipid nanoparticle comprises a particle size that is within about 50% of a particle size of the lipid nanoparticle prior to delivery.

2. A method of delivering a lipid nanoparticle to a subject in need thereof, the method comprising: delivering a formulation comprising the lipid nanoparticle to a targeted region of a nasal cavity of the subject, wherein the formulation is delivered as a liquid jet, wherein the lipid nanoparticle encapsulates an RNA, wherein after a dose of the formulation is delivered, an expression level of the delivered RNA in a cell of the subject is at least about 2-fold an expression level from a same dose of the formulation delivered by a spray method.

3. The method of claim 1 or 2, wherein the method results in an encapsulation efficiency of at least about 2-fold an encapsulation efficiency by a spray method.

4. The method of claim 1 or 2, wherein the delivered lipid nanoparticle comprises a particle size of less than about 100 nm.

5. The method of claim 1 or 2, wherein the delivered lipid nanoparticle comprises a polydispersity index (PDI) of less than about 0.2.

6. The method of claim 1 or 2, wherein the lipid nanoparticle comprises a nucleic acid molecule.

7. The method of claim 1 or 2, wherein the lipid nanoparticle comprises a vaccine.

8. The method of claim 7, wherein the vaccine comprises an RNA vaccine.

9. The method of claim 8, wherein the RNA vaccine comprises a mRNA vaccine.

10. The method of claim 8, wherein the RNA vaccine comprises a saRNA vaccine.

11. The method of claim 8, wherein administering of the formulation results in an increased antigenicity in the subject compared to where the formulation is delivered to the subject as a spray at about the same dose.

12. The method of claim 1, wherein the formulation is delivered by a device, the device comprising:(a) a housing comprising an insertable portion comprising a distal end, and a proximal end; andMintz Ref.: 062706-519001 WO(b) a subject-engaging portion which engages a columella region of the subject to seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject, wherein the device is configured to deliver the formulation as a laminar jet.

13. A method of delivering a delivery vehicle to a subject in need thereof, the method comprising: delivering a formulation comprising the delivery vehicle to a targeted region of a nasal cavity of the subject, wherein the formulation is delivered as a liquid jet.

14. The method of claim 13, wherein the liquid jet is a laminar jet.

15. The method of claim 13, wherein the formulation is delivered as a flow having a Reynold’s number of 2300 or less.

16. The method of claim 13, wherein delivering the formulation as a laminar flow, a liquid jet, or a flow having a Reynold’s number of 2300 or less: a) increases on target delivery of the composition to the target region, b) decreases off target delivery of the composition to the nasal cavity, or c) both, compared to delivering the formulation with a spray ejection profile.

17. The method of claim 13, wherein the formulation is ejected from a dispensing element, wherein at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% of the formulation passes through a circular opening having a diameter of between about 5 mm and about 6 mm at a distance of about 25 mm from where the formulation is ejected from the dispensing element.

18. The method of claim 13, wherein the formulation has a viscosity of between about 0.5 cP and about 100 cP.

19. The method of claim 13, wherein the formulation has a viscosity of between about 0.5 cP and about 50 cP.

20. The method of claim 13, wherein the formulation has a viscosity of between about 0.5 cP and about 10 cP.

21. The method of claim 13, wherein the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 30 m / s.

22. The method of claim 13, wherein the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 5 m / s.

23. The method of claim 13, wherein the formulation is ejected from a dispensing element, wherein at least 75% of the formulation passes through a circular opening having a diameter of between about 5 mm and about 6 mm at a distance of about 25 mm from where theMintz Ref.: 062706-519001 WO formulation is ejected from the dispensing element, wherein the formulation has a viscosity of between about 0.5 cP and about 50 cP, and wherein the formulation is ejected from the dispensing element at a velocity of between about 1 m / s and about 30 m / s.

24. The method of claim 13, wherein at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% of the formulation is delivered to the targeted region of the nasal cavity of the subject.

25. The method of claim 13, wherein the delivery vehicle comprises a macromolecule delivery vehicle.

26. The method of claim 13, wherein the delivery vehicle comprises an exosome.

27. The method of claim 26, wherein the exosome comprises a cargo.

28. The method of claim 27, wherein the cargo is encapsulated by the exosome.

29. The method of claim 1, wherein the delivery vehicle comprises a nucleic acid delivery vehicle.

30. The method of claim 29, wherein the nucleic acid delivery vehicle comprises a viral vector.

31. The method of claim 30, wherein the viral vector comprises an adeno-associated vector (AAV).

32. The method of claim 13, wherein the method results in an encapsulation efficiency at least about 2-fold- at least about 3 -fold, at least about 4-fold, or at least about 5-fold an encapsulation efficiency by a spray method.

33. The method of claim 13, wherein the delivery vehicle comprises a lipid nanoparticle.

34. The method of claim 33, wherein the delivered lipid nanoparticle comprises a particle size of less than about 100 nm.

35. The method of claim 33, wherein the delivered lipid nanoparticle comprises a particle size that is within about 10%, within about 20%, within about 30%, within about 40%, or within about 50% of a particle size of the delivery vehicle prior to delivery.

36. The method of claim 33, wherein the delivered lipid nanoparticle comprises a polydispersity index (PDI) of less than about 0.2.

37. The method of claim 33, wherein the lipid nanoparticle encapsulates an RNA, wherein after a dose of the formulation is delivered, an expression level of the delivered RNA in a cell of the subject is at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, or at least about 3-fold an expression level from a same dose of the formulation delivered by a spray method.

38. The method of claim 33, wherein the lipid nanoparticle comprises a nucleic acid molecule.Mintz Ref.: 062706-519001 WO39. The method of claim 33, wherein the lipid nanoparticle comprises a vaccine.

40. The method of claim 39, wherein the vaccine comprises an RNA vaccine.

41. The method of claim 40, wherein the RNA vaccine comprises a mRNA vaccine.

42. The method of claim 40, wherein the RNA vaccine comprises a saRNA vaccine.

43. The method of claim 33, wherein administering of the formulation results in an increased antigenicity in the subject compared to where the formulation is delivered to the subject as a spray at about the same dose.

44. The method of claim 33, wherein the lipid nanoparticle comprises a cargo.

45. The method of claim 44, wherein the cargo is encapsulated by the lipid nanoparticle.

46. The method of claim 44, wherein the cargo comprises a nucleic acid molecule, a small molecule, a protein, a peptide, or any combination thereof.

47. The method of claim 13, wherein the targeted region is an olfactory region.

48. The method of claim 13, wherein the targeted region is a turbinate region.

49. The method of claim 13, wherein the turbinate region is a middle turbinate region.

50. The method of claim 13, wherein the targeted region is a nasal associated lymphatic tissue.

51. The method of claim 13, wherein the targeted region is a nasopharynx region.

52. The method of claim 13, wherein the formulation is delivered by a device, the device comprising:(a) a housing comprising an insertable portion comprising a distal end, and a proximal end; and(b) a subject-engaging portion which engages a columella region of the subject to seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject, wherein the device is configured to deliver the formulation as a laminar jet.

53. The method of claim 52, wherein the ejection zone is:(a) 0mm to 30mm superior to a horizontal line that intersects the anterior aspect of the internal nasal valve, and(b) 0mm to 20mm anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule.

54. The method of claim 53, wherein the ejection zone is further: i. 0mm to 40mm inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft,Mintz Ref.: 062706-519001 WO ii. Omm to 20mm posterior to the internal nasal dorsum, iii. 10mm to 50mm superior to a horizontal line that intersects the inferior aspect of the columella, iv. Omm to 30mm superior to a horizontal line that intersects the superior aspect of the inferior turbinate, v. Omm to 3 mm from the septum, or vi. any combination thereof.

55. A device for intranasal delivery of a delivery vehicle to a targeted region of a nasal cavity of a subject, the device comprising: a) a housing comprising an insertable portion comprising a distal end, and a proximal end; and b) a subject-engaging portion which engages a columella region of the subject to seat the distal end of the insertable portion within an ejection zone of a nasal channel of the subject; wherein the device is configured to deliver a formulation comprising the delivery vehicle to the targeted region of the subject.

56. The device of claim 55, wherein the device dispenses the formulation as a laminar jet.

57. The device of claim 55, configured to deliver at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% of the formulation to the targeted region of the nasal cavity of the subject.

58. The device of claim 55, wherein the device comprises a compliant dispensing tip comprising a compliant and flexible soft nib.

59. The device of claim 55, wherein the dispensing tip has a diameter of between about 1.2 mm and 1.4 mm.

60. The device of claim 55, wherein the dispensing tip has a diameter of about 1.3 mm.

61. The device of claim 55, wherein the dispensing tip has a diameter of about 1.35 mm.

62. The device of claim 55, wherein the formulation has a viscosity of between about 0.5 cP and about 100 cP.

63. The device of claim 55, wherein the device is configured to deliver the formulation at a velocity of between about 1 m / s and about 30 m / s.

64. The device of claim 55, wherein the device is configured to deliver the formulation at a velocity of between about 1 m / s and about 5 m / s.Mintz Ref.: 062706-519001 WO65. The device of claim 55, wherein application of pressure by the subject-engaging portion to the columella region of the subject enables and / or causes delivery of the formulation to the subject from the insertable portion.

66. The device of claim 55, wherein the insertable portion comprises a dispensing element for delivery of the formulation to the targeted region of the subject.

67. The device of claim 55, wherein the ejection zone is:(a) Omm to 30mm superior to a horizontal line that intersects the anterior aspect of the internal nasal valve, and(b) Omm to 20mm anterior to an inclined line that intersects the anterior aspect of the middle turbinate and the posterior aspect of the vestibule.

68. The device of claim 67, wherein the ejection zone is further: i. Omm to 40mm inferior to a horizontal line that is parallel to the inferior aspect of the olfactory cleft, ii. Omm to 20mm posterior to the internal nasal dorsum, iii. 10mm to 50mm superior to a horizontal line that intersects the inferior aspect of the columella, iv. Omm to 30mm superior to a horizontal line that intersects the superior aspect of the inferior turbinate, v. Omm to 3 mm from the septum, or vi. any combination thereof.

69. The device of claim 55, configured to deliver a lipid nanoparticle comprising a particle size of less than about 100 nm post-delivery.

70. The device claim 55, configured to deliver a lipid nanoparticle comprising a particle size that is within about 10%, within about 20%, within about 30%, within about 40%, or within about 50% of a particle size of the lipid nanoparticle prior to delivery.

71. The device of claim 55, configured to deliver a lipid nanoparticle comprising a polydispersity index (PDI) of less than about 0.2.

72. The device of claim 55, configured to deliver the delivery vehicle with an encapsulation efficiency at least about 2-fold- at least about 3-fold, at least about 4-fold, or at least about 5- fold an encapsulation efficiency by a spray device.

73. The device of claim 55, configured such that when the delivery vehicle comprises a lipid nanoparticle encapsulating an RNA, after a dose of the formulation is delivered, anMintz Ref.: 062706-519001 WO expression level of the delivered RNA in a cell of the subject is at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, or at least about 3-fold an expression level from a same dose of the formulation delivered by a spray device.

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