Method and adapter for mixing pharmaceutical conjugates - Patent Application 20070122997

Abstract

An adapter for mixing a first substance and a second substance, a system including the same, and a method for using the same are provided. The adapter may include a body and a mixing pin. The body may have a first port configured to connect to a first container and receive a first substance, a second port configured to connect to a second container and receive a second substance, a third port configured to connect to the receiving container and output a mixture of both substances, and a mixing chamber. The mixing chamber includes a first portion communicating with the first and second ports and a second portion communicating with the third port. The mixing pin may be inserted into the mixing chamber and at least partially define at least one microfluidic pathway for mixing the first and second substances. At least one of the first channel and the second channel may be oriented offset from the midplane of the body.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This patent application claims priority to U.S. Provisional Patent Application No. 63 / 378230, filed October 3, 2022, the entire disclosure of which is expressly incorporated herein by reference.

[0002] The present disclosure relates to adapters, systems, and methods of use for mixing at least two substances to form a pharmaceutical conjugate. [Background technology]

[0003] Recent developments in immunology include the newly approved messenger RNA-encapsulated lipid nanoparticle (mRNA-LNP) vaccine. Messenger RNA (mRNA) technology has the advantage of being able to rapidly adapt to new antigen designs by modifying the mRNA sequence without the need to overhaul the vaccine's chemical and manufacturing controls (CMC). However, mRNA provided alone is not easily absorbed or effectively delivered to human immune cells, has unstable chemical and physical properties, and is therefore ineffective for use as a vaccine. Recent developments have shown that mRNA absorption and stability can be increased to effective levels when encapsulated within lipid nanoparticle (LNP) vectors.

[0004] Preparation of mRNA-LNP vaccines is achieved by mixing lipid-dissolved ethanol with RNA in a buffer solution under strictly controlled conditions. Such mixing is typically performed in laboratories using various equipment, but these are often unsuitable for large-scale distribution due to their low durability, high cost, high complexity, poor lot-to-lot consistency, and / or high batch-to-batch variability. Summary of the Invention

[0005] The inventors have recognized that mRNA-LNP vaccines have a limited shelf life at room temperature. To extend their shelf life, mRNA-LNP vaccines must be stored at extremely low temperatures (typically -20 to -80 degrees Celsius). This is problematic because low-temperature distribution is expensive and logistically complex. In addition, there is a risk that mRNA-LNP vaccines will be wasted if, for example, the low-temperature environment at any stage of the distribution chain becomes inoperable.

[0006] Non-messenger RNA drugs, such as RNAi, siRNA, and other oligonucleotides, can also be formulated into lipid nanoparticle compositions (RNA-LNPs). RNA-LNP drugs can be chemically modified to improve their stability and shelf life at room temperature (such chemical modifications are not possible with mRNA-LNP technology, which requires interaction with cellular proteins to function properly). Chemical modification of RNA-LNPs can be difficult and expensive to achieve, but is nonetheless often preferred to avoid the significant distribution costs associated with unmodified RNA-LNP drugs, which must also be kept at very low temperatures, as well as the difficulties associated with managing drug efficacy over time due to their limited molecular half-life.

[0007] In short, the low temperature requirement presents a major challenge for distribution and development. Other problems associated with known systems for producing nanoparticle compositions include limited scalability, usability, and / or reliability. One or more of the aforementioned needs are met by various embodiments as disclosed herein.

[0008] A first aspect of the present disclosure relates to an adapter having a body having a first port configured to connect with a first container to receive a first substance, a second port configured to connect with a second container to receive a second substance, a third port configured to connect with a receiving container to output a mixture of the first substance and the second substance, a mixing chamber having a first portion communicating with the first port and the second port and a second portion communicating with the third port, and a mixing pin inserted into the mixing chamber, the mixing pin at least partially defining at least one microfluidic pathway for mixing the first substance and the second substance.

[0009] The adapter may have one or more of the following features. The body may further include an opening in the first portion of the mixing chamber configured to receive the mixing pin. The mixing pin may be removable from the mixing chamber. The mixing pin may have a flange, and the body may have a recess configured to receive the flange. The mixing pin may have a sealing portion configured to seal the first portion of the mixing chamber. The first port and / or the second port may be substantially parallel to the mixing chamber. The body may further include a first branch channel communicating with the first port and extending at a first angle relative to the mixing chamber, and a second branch channel communicating with the second port and extending at a second angle relative to the mixing chamber. The first angle and / or the second angle may be greater than 90 degrees. The first angle and / or the second angle may be between about 120 degrees and about 160 degrees. The adapter may further include a first plug inserted into the end of the first branch channel and a second plug inserted into the end of the second branch channel. The first branch channel may be configured to guide the first substance toward the mixing pin, and the second branch channel may be configured to guide the second substance toward the mixing pin. The mixing pin defines a first edge surface, which may be formed by a first surface and a second surface, and a second edge, which may be formed by a third surface and a fourth surface, wherein the first edge is configured to divide the first substance and the second edge is configured to divide the second substance. The first port may be a female Luer connector, the second port may be a female Luer connector, and the third port is a male Luer connector. The at least one microfluidic pathway may be serpentine. The at least one microfluidic pathway may be formed by at least one channel. The at least one channel may have a dimension of about 200 μm to about 1000 μm. The at least one channel may have a dimension of about 400 μm to about 600 μm. The at least one channel may include at least one spiral channel.The at least one helical channel may include a first helical channel and a second helical channel, the first helical channel and the second helical channel intersecting at multiple intersections along the longitudinal length of the mixing pin. The at least one channel may extend through the mixing pin. The mixing pin may include a plurality of protrusions extending at least partially around the circumference of the mixing pin and defining at least a portion of the at least one channel. The plurality of protrusions may extend uniformly around the circumference of the mixing pin and form at least one radial gap with the inner surface of the body, the at least one radial gap forming at least a portion of the at least one channel. Each of the plurality of protrusions may have at least one first surface configured to form a seal with the inner surface of the body and at least one second surface forming the at least one radial gap with the inner surface of the body, the at least one radial gap forming at least a portion of the at least one channel.

[0010] A second aspect of the present disclosure relates to a system comprising: an aqueous solution; a lipid solution; a first container configured to receive the aqueous solution; a second container configured to receive the lipid solution; a receiving container; and an adapter, the adapter comprising: a body having a first port connected to the first container and configured to receive the aqueous solution; a second port connected to the second container and configured to receive the lipid solution; a third port connected to the receiving container and configured to output a mixture of the aqueous solution and the lipid solution; and a mixing chamber extending between a first portion communicating with the first port and the second port and a second portion communicating with the third port; and a mixing pin configured to be inserted into the mixing chamber, the mixing pin at least partially defining at least one microfluidic path for mixing the aqueous solution and the lipid solution.

[0011] The system may have one or more of the following features: The system may include a second pin configured to be inserted into the mixing chamber, the second mixing pin having a different geometric shape than the mixing pin; The system may include a first storage container containing an aqueous solution and a second storage container containing a lipid solution.

[0012] A third aspect of the present disclosure relates to a method including introducing a first substance into a first port of an adapter; introducing a second substance into a second port of the adapter; mixing the first substance and the second substance in at least a microfluidic path through, along, and / or around a mixing pin in the adapter; and producing a pharmaceutical complex of the first substance and the second substance.

[0013] The method may have one or more of the following features: the first substance may be an aqueous solution, the second substance may be a lipid solution, and the pharmaceutical complex may comprise lipid nanoparticles.

[0014] A fourth aspect of the present disclosure relates to an adapter including a body having a first port configured to connect with a first container to receive a first substance, a first channel in communication with the first port, a second port configured to connect with a second container to receive a second substance, a second channel in communication with the second port, a third port configured to connect with a receiving container to output a mixture of the first substance and the second substance, a mixing chamber having a first portion in communication with the first port and the second port and a second portion in communication with the third port, and a mixing pin inserted into the mixing chamber, wherein at least one of the first channel and the second channel is oriented offset from the midplane of the body.

[0015] The adapter may have one or more of the following features. The body may include an opening in a first portion of the mixing chamber configured to receive the mixing pin. The mixing pin may be removable from the mixing chamber. The mixing pin may have an end cap, and the body may have a rim configured to engage the end cap. The adapter may include a sealing member configured to seal the first portion of the mixing chamber. The first port and / or the second port may be substantially parallel to the mixing chamber. The first channel may extend at a first angle relative to the mixing chamber, and the second channel may extend at a second angle relative to the mixing chamber. The first angle and / or the second angle may be greater than 90 degrees. The first angle and / or the second angle may be from about 120 degrees to about 160 degrees. The adapter may include a first plug at an end of the first channel and a second plug at an end of the second channel. The first channel and the second channel may be offset from a midplane of the mixing pin. The first channel and the second channel may be on the same side of the horizontal axis of the adapter body. The first channel and the second channel may be on opposite sides of the horizontal axis of the adapter body. The first channel and the second channel may be connected to the body at different longitudinal positions. The first port may be a female Luer connector, the second port may be a female Luer connector, and the third port may be a male Luer connector. The mixing pin may form at least one channel. The at least one channel may have a dimension of about 200 μm to about 1000 μm. The at least one channel may have a dimension of about 400 μm to about 600 μm. The at least one channel may include at least one spiral channel.

[0016] A fifth aspect of the present disclosure relates to a system comprising: an aqueous solution; a lipid solution; a first container configured to receive the aqueous solution; a second container configured to receive the lipid solution; a receiving container; an adapter, the adapter having a body having a first port connected to the first container and configured to receive the aqueous solution; a second port connected to the second container and configured to receive the lipid solution; a third port connected to the receiving container and configured to output a mixture of the aqueous solution and the lipid solution; a mixing chamber extending between a first portion communicating with the first port and the second port and a second portion communicating with the third port; and a mixing pin configured to be inserted into the mixing chamber, wherein at least one of the first channel and the second channel is oriented offset from the midplane of the body.

[0017] The system may have one or more of the following features: The system may include a second pin configured to be inserted into the mixing chamber, the second mixing pin having a different geometric shape than the mixing pin; The system may include a first storage container containing an aqueous solution and a second storage container containing a lipid solution.

[0018] A sixth aspect of the present disclosure relates to a method including: introducing a first substance into a first port of a body of an adapter through a first channel of the body of the adapter; introducing a second substance into a second port of the body of the adapter through a second channel of the body of the adapter; mixing the first substance and the second substance around a mixing pin in the body of the adapter, wherein at least one of the first channel and the second channel is oriented offset from a midplane of the body of the adapter; and producing a pharmaceutical complex of the first substance and the second substance.

[0019] The method may have one or more of the following features: the first substance may be an aqueous solution, the second substance may be a lipid solution, and the pharmaceutical complex may comprise lipid nanoparticles. [Brief explanation of the drawings]

[0020] Particular embodiments of the present disclosure are described, by way of example only, in the following detailed description and with reference to the accompanying drawings. [Figure 1] FIG. 1 illustrates a side view of an exemplary system according to the present disclosure. [Figure 2] 2 shows an exploded view of a first embodiment of the adapter of the system of FIG. 1. [Figure 3] 3 shows a first cross-sectional view of the adapter of FIGS. 1 and 2; FIG. [Figure 4] 4 shows a second cross-sectional view of the adapter of FIGS. 1 to 3. FIG. [Figure 5] FIG. 5 illustrates a first side view of the mixing pin of the adapter of FIGS. [Figure 6] FIG. 6 illustrates a second side view of the mixing pin of the adapter of FIGS. 1 to 5. [Figure 7] FIG. 7 illustrates a third side view of the mixing pin of the adapter of FIGS. 1 to 6. [Figure 8] 5 shows a cross-sectional view of a second embodiment of a mixing pin in the adapter of the system of FIGS. 1 to 4. FIG. [Figure 9] 9 shows a cross-sectional view of the adapter of FIG. 8. [Figure 10] FIG. 5 shows an isometric view of a third embodiment of the mixing pin of the system of FIGS. [Figure 11] 11 shows a cross-sectional view of a third embodiment of the mixing pin of FIG. 10 in the adapter of FIGS. 1 to 4. FIG. [Figure 12] 12 shows a liquid volume cross section of the adapter of FIG. 11. [Figure 13] FIG. 5 shows an isometric view of a fourth embodiment of the mixing pin of the system of FIGS. [Figure 14] 5 shows a partial perspective view of a second embodiment of the adapter of the system of FIGS. 1 to 4. FIG. [Figure 15] 5 shows a partial perspective view of a third embodiment of the adapter of the system of FIGS. 1 to 4. FIG. [Figure 16] 16 shows a longitudinal view of the adapter of FIG. 15. [Figure 17] 17 shows a cross-sectional view of the adapter of FIGS. 15 and 16. FIG. [Figure 18]FIG. 18 shows an isometric view of the adapter of FIGS. 15 to 17. [Figure 19] 19 shows an exploded view of the adapter of FIGS. 15 to 18. FIG. [Figure 20] FIG. 20 shows an isometric view of the adapter of FIGS. 15 to 19. [Figure 21] FIG. 20 shows a first isometric view of the mixing pin of FIGS. 15 to 17 and 19. [Figure 22] FIG. 20 shows a second isometric view of the mixing pin of FIGS. 15-17 and 19. [Figure 23] FIG. 20 shows an isometric view of the first plug of the adapter of FIGS. 15 to 19; [Figure 24] FIG. 20 shows an isometric view of the second plug of the adapter of FIGS. 15 to 19; [Figure 25] FIG. 5 shows an isometric view of a fourth embodiment of the adapter of the system of FIGS. 1 to 4. [Figure 26] 26 shows a longitudinal view of the adapter of FIG. 25. [Figure 27] 27 shows a first cross-sectional view of the adapter of FIGS. 25 and 26. FIG. [Figure 28] 27 shows a second cross-sectional view of the adapter of FIGS. 25 and 26. FIG. [Figure 29] 20 illustrates a method of mixing a first substance and a second substance using one or more of the embodiments of FIGS. 1-28.

[0021] The same reference numbers are used in the drawings and the following detailed description to refer to the same or similar parts. DETAILED DESCRIPTION OF THE INVENTION

[0022] An adapter for connecting one or more fluid containers for microfluidic mixing of first and second substances to produce a pharmaceutical conjugate is provided. The adapter includes a first port or connector member configured to connect to the first container, a second port or connector member configured to connect to the second container, and a third port or connector member configured to connect to a receiving container. The adapter further includes a mixing chamber extending from a first portion in fluid communication with the first and second ports to a second portion in fluid communication with the third port. A mixing pin may be received within the mixing chamber and may at least partially define at least one microfluidic pathway through, longitudinally along, and / or circumferentially around the mixing pin. The at least one pathway can induce turbulence in the fluid flow through the mixing chamber to mix the fluid components as they flow through the adapter to the receiving container.

[0023] In some embodiments, at least one path may be serpentine and configured to induce local changes in the flow direction of the components flowing through the mixing chamber. In some embodiments, at least one path may include multiple paths. In some embodiments, the multiple paths may have multiple intersections configured to induce local changes in the flow direction of the liquid moving through the mixing chamber. In some embodiments, at least one path may have a variable width or diameter configured to cause repeated acceleration and deceleration, thus inducing turbulence. In some embodiments, at least one path may generate vortices to create highly turbulent mixing.

[0024] The adapters of the present disclosure may offer an improvement over planar microfluidic chips by creating a three-dimensional microfluidic template for mixing. Three-dimensional microfluidic templates as discussed herein enable improved mixing by increasing volume and flow rate while reducing fluid pressure in the channels. For example, the adapters may have larger microfluidic channels or flow paths (e.g., about 400 μm to about 1000 μm) than microfluidic chips and / or provide multiple flow paths that may converge along the length of the adapter, increasing mixing. Thus, the adapters may have larger features that are easier and / or cheaper to manufacture, for example, through injection molding or three-dimensional printing. The adapters can fill a large gap in the LNP synthesis space. Various embodiments of at least one pathway may provide an easy-to-use, consistent, safe, and / or convenient way to mix components at the point of care. The adapters may be used as manual devices (e.g., with a syringe) in small-scale settings or in automated configurations targeting a broad customer segment working in early preclinical stages to potentially scale up manufacturing.

[0025] 1 and 2 show a system 10 including a first container 20, a second container 40, a receiving container 60, an adapter 100, and a mixing pin 160. In some embodiments, the system 10 may further include a first storage container 80 containing a first substance or component and a second storage container 82 containing a second substance or component. The first container 20 may be configured to transfer the first substance from the first storage container 80 to the adapter 100. The second container 40 may be configured to transfer the second substance from the second storage container 82 to the adapter 100. The adapter 100 may include at least one pathway configured to mix the first and second substances and transfer the pharmaceutical complex to the receiving container 60. System 10 may constitute a kit of at least one or all of first container 20, second container 40, receiving container 60, adapter 100, mixing pin 160, first storage container 80, and / or second storage container 82. System 10 and / or kit may further include one or more vial adapters 70 for fluid transfer to or from one or more of receiving container 60, first storage container 80, and / or second storage container 82. Kit components may include packaging for shipment to an end user. While shown with adapter 100 and mixing pin 160, system 10 and / or kit may include any one or more of adapters 100, 500, 600, 700 and / or mixing pins 160, 260, 360, 460, 660, 760. In some embodiments, the system 10 and / or kit may include a single adapter and multiple mixing pins 160, 260, 360, 460, 660, 760 of the same or different geometries. For example, multiple different types of mixing pins 160, 260, 360, 460, 660, 760 may be included in the system 10 and / or kit to allow an end user to select or interchange the mixing pins 160, 260, 360, 460, 660, 760 based on the intended purpose and / or components.

[0026] The first container 20 may be a variable volume container, such as a first syringe, configured to at least temporarily store and / or transfer a first substance from the first storage container 80 to the adapter 100. The second container 40 may be a variable volume container, such as a second syringe, configured to store and transfer a second substance from the storage container 82 to the adapter 100. The first syringe 20 may include a first syringe body 22 and a first plunger rod 24, and the second syringe 40 may include a second syringe body 42 and a second plunger rod 44. Each syringe body 22, 42 may have a syringe barrel extending longitudinally from a proximal end to a distal end. Each syringe body 22, 42 may have a syringe tip at the distal end and a flange at the proximal end. The syringe barrel may be tubular with an inner surface extending longitudinally to define a chamber. The chambers may be configured to receive, store, and / or mix a substance for dispensing through the distal opening of the syringe tip. The first plunger rod 24 may have a first flange 25 at its proximal end, and the second plunger rod 44 may have a second flange 45 at its proximal end. The syringe tip of the first syringe body 22 may include a first connector 26 for engaging an external device, such as a syringe needle, a container, and / or an adapter 100. The syringe tip of the second syringe body 42 may include a second connector 46 for engaging the same or a different external device, such as a syringe needle, a container, and / or an adapter 100. Each connector 26, 46 may further include a male Luer connector including a syringe tip and a threaded sleeve around the tip. The syringe tips may be tapered to guide fluid flow into an external device (e.g., adapter 100), and the sleeves may have internal threads configured to secure syringes 20, 40 to their respective external devices (e.g., adapter 100). Reservoirs 20, 40 may be any conventional syringe and / or reciprocating pump suitable for use in a pharmaceutical environment.

[0027] The flanges 25, 45 may be actuated either by being pulled to create a negative pressure to draw the substance into the chamber and / or by being pushed to create a positive pressure to expel the substance from the chamber. At least a portion of the first syringe 20 and the second syringe 40 may be integrally or releasably connected to enable coordinated manipulation and / or actuation of the first syringe 20 and the second syringe 40. For example, the system 10 may further include a barrel holder (not shown) having a first lumen configured to receive the first syringe body 22 and a second lumen configured to receive the second syringe body 42 so that the first syringe 20 and the second syringe 40 may be manipulated together. Each of the first and second lumens may be closed or formed by a C-shaped wall configured to snap around the respective syringe body 22, 42. The barrel holder may secure the syringe bodies 22, 42 in a substantially parallel arrangement. The system may further include a plunger clip configured to translate the plunger rods 24, 44 together through the syringe bodies 22, 42 to push and / or pull the material in the same longitudinal translation. For example, the plunger clip may be configured to attach to the flanges 25, 26 and may have grooves configured to releasably receive the flanges 25, 45. Embodiments of barrel holders and / or plunger clips are further discussed in U.S. Patent Nos. 5,104,375, 6,840,921, and 8,240,511, the entire disclosures of which are expressly incorporated herein by reference.

[0028] The receiving vessel 60 may be a fixed volume container, such as a vial, that can be attached to the adapter 100 using the vial adapter 70. The vial 60 may include a vial bottle 62 having a crown and a neck enclosing a chamber. The chamber may be sealed by a drug vial seal at the crown, which is circumferentially attached by an aluminum band. The vial adapter 70 may have a transverse top wall 72, a connector 74 extending upwardly from the top wall 72, and a skirt 76 extending downwardly from the top wall 72. The connector 74 may be a female Luer connector including external threads for screw-thread engagement with a male Luer lock connector, such as the connector of the adapter 100. The skirt 76 may be for telescopic attachment over the crown and / or neck of the vial 60. The skirt 76 may enclose a cannula (not shown) extending downwardly from the top wall 72 and configured to pierce a vial stopper. The cannula may have a lumen that fluidly communicates with the chamber of the vial bottle 62 when puncturing the vial stopper. The vial adapter 70 may be vented to draw air into the container 60 and facilitate drawing fluid through the system. Further discussion of embodiments of the container 60 and / or vial adapter 70 is provided in U.S. Pat. Nos. 8,753,325 and 9,943,463, the entire disclosures of which are expressly incorporated herein by reference. The receiving container 60 may be configured to initially contain a buffer solution and to receive materials dispensed from the first and second containers 20, 40 and mixed within the adapter 100. After the first and second components are introduced into the adapter 100, the resulting pharmaceutical complex may be stored in the receiving container 60.

[0029] However, in some embodiments, the receiving container 60 may be a variable volume container, such as a syringe, and one or both of the first and second containers 20, 40 may be fixed volume containers, such as vials. Further discussion of such embodiments is provided in U.S. Patent Application Publication No. 2023 / 0105059, the entire disclosure of which is expressly incorporated herein by reference. It is also contemplated that the first and second containers 20, 40 may be embodied as first and second containers 20, 40 connected to pumps. Flow sensors may be connected to the first and second containers 20, 40 to control the flow rates of the first and second substances.

[0030] The first storage container 80 and / or the second storage container 82 may have a structure similar to the receiving container 60, the discussion of which is expressly incorporated herein in its entirety. For example, each of the first and second storage containers 80, 82 may be a fixed volume container enclosing a chamber and having a crown 84 and a neck 85. The chamber may be sealed by a drug vial seal 86 at the crown 84, which is circumferentially attached by an aluminum band. Each of the first and second storage containers 80, 82 may be attached to the vial adapter 70, as discussed with reference to the receiving container 60.

[0031] The first substance in first storage container 80 may be an aqueous solution. The aqueous solution may be any aqueous buffer solution that can be used to dissolve nucleic acids. For example, in some embodiments, the aqueous solution may be a solution of 20 mM citrate and 300 mM sodium chloride and may have a pH ranging from 3 to 6. In some embodiments, the aqueous solution may be 20 mM phosphate buffer (PBS) at pH 7. In some embodiments, the aqueous solution may be a solution of 5 mM to 25 mM sodium acetate buffer at a pH range of 4 to 6.

[0032] The second substance in the second storage container 82 may be a lipid solution having a composition comprising, in whole or in part, an organic solvent containing a lipid or a mixture of lipids. The lipid solution may comprise clinical-grade lipids solubilized in an organic alcohol solution (e.g., ethanol). In some embodiments, the lipid solution may be at least a 25% alcohol solution. In some embodiments, the lipid solution may be at least a 40% alcohol solution. In some embodiments, the lipid solution may be at least a 60% alcohol solution. The alcohol solution is preferably an ethanol solution. Providing lipids in such an increased alcohol concentration may allow the lipids in the alcohol solution to withstand dilution by a reconstitution agent without affecting the quality of the resulting pharmaceutical conjugate. The lipid composition in the ethanol solution may be composed of ionizable or cationic lipids or synthetic lipids, structured lipids, PEG-lipids or derivatives thereof, and cholesterol or derivatives thereof. However, the second substance may also comprise other nanoparticle-forming solutions.

[0033] A therapeutic agent may be carried in at least one of the first substance and / or the second substance. In a preferred embodiment, the therapeutic agent is carried in the first substance. The therapeutic agent may include a nucleic acid, a drug, a protein, an oligonucleotide, etc., including a gene editing complex. The nucleic acid may include RNA and / or DNA. The RNA may be in the form of an oligonucleotide RNA, tRNA (transfer RNA), snRNA (small nuclear RNA), rRNA (ribosomal RNA), mRNA (messenger RNA), antisense RNA, siRNA (small interfering RNA), shRNA (short hairpin RNA), ncRNA (non-coding RNA), an aptamer, a ribozyme, a chimeric sequence, or derivatives of these groups. The gene editing complex may include a gRNA (guide RNA), a cas9 protein, an mRNA or DNA encoding the cas9 protein, or a CRISPR-cas9gRNA complex. The DNA may be in the form of antisense, plasmid DNA, a portion of plasmid DNA, pre-condensed DNA, a polymerase chain reaction (PCR) product, a vector (P1, PAC, BAC, YAC, artificial chromosome), an expression cassette, a chimeric sequence, chromosomal DNA, or derivatives of these groups.

[0034] In some embodiments, the therapeutic agent may be stored in a dehydrated and / or lyophilized state and reconstituted in an aqueous solution to form the first substance before being introduced into adapter 100. In this case, first storage container 80 may hold the dehydrated therapeutic agent, and first container 20 may hold the aqueous solution. The aqueous solution may then be introduced from first container 20 into first storage container 80 to reconstitute the therapeutic agent. The first solution containing the therapeutic agent may then be introduced into first container 20 and introduced into the adapter.

[0035] The first substance (with the therapeutic agent in a lyophilized state or as a solution) and the second substance may be adapted for transport and medium- or long-term storage at room temperature. Thus, the system 10 and adapter 100 disclosed herein may enable mitigation of the obstacles associated with storing and transporting RNA-LNP complexes at very low temperatures. Furthermore, the adapter 100 may be easy to use in clinical settings.

[0036] The first and second substances may be mixed using an adapter, as discussed herein. Mixing may produce a liposome former, which simultaneously forms liposomes and entraps the therapeutic agent. Electrostatic interactions between a negatively charged therapeutic agent (e.g., nucleic acid or mRNA) and a positively charged cationic lipid may form LNPs that encapsulate the therapeutic agent. If mRNA is included in the mixture, the LNPs may encapsulate the mRNA to form mRNA-LNPs. The pharmaceutical complex may be a monodisperse LNP. Thus, a ready-to-inject RNA-LNP (e.g., mRNA-LNP) complex may be formed by mixing components of an aqueous solution containing RNA and a lipid solution using the system 10 and adapter 100. Considering the specific example of forming an mRNA-LNP pharmaceutical complex, the complex may be formed by mixing a first substance containing mRNA (or RNA) from a first container 20 with a second substance in a lipid solution from a second container 40 via any of the adapters disclosed herein.

[0037] The first and second storage containers 80, 82 may be the same or different sizes depending on the intended mixture. Accordingly, any reference herein to two storage containers should be interpreted as including three or more storage containers. It will be understood that where three or more components are to be mixed, three or more containers may be provided, each containing at least one component. Furthermore, any number of components may be provided unmixed in a single container. In some embodiments, the first container 20 and the second container 40 may be pre-filled with the respective substances, e.g., embodied as pre-filled syringes, and the first and second storage containers 80, 82 may be omitted.

[0038] As further shown in Figures 2-4, the adapter 100 may include a body 101 having a first port 102 configured to connect to the syringe connector 26 of the first container 20 and a second port or connector member 104 configured to attach to the syringe connector 46 of the second container 40.

[0039] The first port 102 may be configured to be received by the first syringe connector 26 and may have external threads 106 configured to mate with the internal threads of the first connector 26. Similarly, the second port 104 may be configured to be received by the second connector 46 and may have external threads 108 about the second tubular member 104 configured to mate with the internal threads of the second syringe connector 46. For example, the ports 102, 104 may be female Luer connectors, and the connectors 26, 46 of the containers 20, 40 may be male Luer connectors. However, the ports 102, 104 may additionally or alternatively connect to the containers 20, 40 with other types of connections, such as snap-fit, friction-fit, and / or press-fit. The adapter 100 may be configured to connect to any number of first and second containers 20, 40, and the adapter 100 may therefore have an equal number of ports 102, 104 for connecting to each of the containers 20, 40. Additionally, one or more of the ports 102, 104 may include a one-way valve (not shown) to allow fluid flow from the container 20, 40 to the adapter 100 and to restrict or substantially prevent fluid flow from the adapter 100 back to the respective container 20, 40.

[0040] The first port 102 may define a first connector channel or port 110, and the second connector member 104 may define a second connector channel 112. In some embodiments, the first connector channel 110 may be configured to receive a tip of the first syringe 20 to place a chamber of the first syringe body 22 in fluid communication with the first connector channel 110, and the second connector channel 112 may be configured to receive a tip of the second syringe 40 to place a chamber of the second syringe body 42 in fluid communication with the second connector channel 112. The first tubular member 102 and the second tubular member 104 may extend substantially parallel to one another to facilitate coordinated actuation of the syringes 20, 40, as discussed herein. In some embodiments, as shown in FIGS. 1-4, the first tubular member 102 and the second tubular member 104 may extend substantially parallel to the longitudinal axis of the adapter body 101.

[0041] The adapter body 101 may further include a first branch member 114, a second branch member 116, and a mixing member 118. The first port 102 may extend from the first branch member 114, and the first branch member 114 may connect the first port 102 to the mixing member 118. The second port 104 may extend from the second branch member 116, and the second branch member 116 may connect the second port 104 to the mixing member 118. The first branch member 114 may have a first branch channel 120 configured to receive a fluid flow from the first container 20 through the first connector channel 110, and the second branch member 116 may have a second branch channel 122 configured to receive a fluid flow from the second container 40 through the second connector channel 112. The first and second branch channels 120 and 122 may communicate with a first or upper portion of the mixing chamber 124. The first and second branch channels 120 and 122 may be angled relative to a longitudinal axis of the mixing element 118, the adapter body 101, and / or the mixing pin 160 to converge fluid flow from the first and second syringes 20 and 40 into the mixing element 118. The angle of at least one of the first and second branch channels 120 and 122 may be at least 90 degrees relative to the mixing element 118, the adapter body 101, and / or the mixing pin 160. As further shown, the angle of at least one of the first and second branch channels 120 and 122 may be from about 120 degrees to about 160 degrees relative to the mixing element 118, the adapter body 101, and / or the mixing pin 160. The first branch channel 120 and the second branch channel 122 may form a Y-shape with the mixing element 118. This configuration of the branch channels 120, 122 and the ports 102, 104 may space the syringes 22, 42 far enough apart to facilitate manual mixing and allow the syringes to engage on the adapter body 101 and actuate together. However, other configurations may be used for other applications. For example, the ports 102, 104 may be closer together if configured to attach to the microtubing of a pump.

[0042] The first branch channel 120 and the second branch channel 122 may be aligned with the midplane of the adapter body 101, and the first branch channel 120 and the second branch channel 122 may direct the first and second substances toward opposing faces of the mixing pin 160 and / or create a collision between the two fluids for initial mixing. In some embodiments, the first branch channel 120 and the second branch channel 122 may have the same or substantially the same width or diameter (as shown in FIGS. 3 and 8 ) for applications where the same flow rate is desired for the first and second substances. In some embodiments (as shown in FIGS. 11 and 12 ), the first branch channel 120 and the second branch channel 122 may have different widths or diameters for applications where different flow rates are desired for the first and second substances. For example, the first branch channel 120 may have a larger width or diameter than the second branch channel 122 (as further shown in FIGS. 11 and 12 ). In some embodiments (as shown in FIGS. 2, 3, and 8), at least one or both of the first branch channel 120 and the second branch channel 122 may be formed with an open outer end, and the adapter 100 may include a plug 140 inserted into one or both of the branch members 114, 116 to seal the outer end of the respective branch channel 120, 122. In some embodiments (as shown in FIG. 11), at least one or both of the first branch channel 120 and the second branch channel 122 may have a closed outer end formed by the adapter body 101.

[0043] The plug 140 may be removable, replaceable, interchangeable, and / or adjustable. The plug 140 may facilitate manufacturing of the adapter body 101 by allowing the branch channels 120, 122 to be formed in the adapter body 101 through injection molding by inserting a straight pin (not shown). The pin may be removed from the injector body 101 after being formed through the opening, and the plug 140 may be inserted to seal the opening in the branch channels 120, 122. In some embodiments, the plug 140 may extend further through the branch channels 120, 122 to reduce the negative space within the branch channels 120, 122. For example, as shown in FIGS. 23 and 24 , the plug 140 may include a pin 642 that occupies space within the branch channels 120, 122 to reduce the negative space. In this manner, the inventors have discovered that the volume of the branch channels 120, 122 may be reduced or optimized without creating manufacturing complexities in injection molding and / or three-dimensional printing of small components within the adapter 101. Furthermore, replacing and / or adjusting the plug 140 may change the inlet volume of one or both of the first connector channel 110 and / or the second connector channel 112. For example, the plug 140 may be replaced with a similar plug 140 having a different length and / or geometry to at least partially occlude one or both of the first connector channel 110 and / or the second connector channel 112. In some embodiments, the plug 140 may be adjusted by pushing or translating it into the branch channels 120, 122 to at least partially occlude the first connector channel 110 and / or the second connector channel 112. In some embodiments, plug 140 may have an adjustable length, such as being telescoping, to at least partially occlude first connector channel 110 and / or second connector channel 112. Varying the inlet volume with plug 140 may adjust the amount of fluid mixing and / or reduce the dead volume of fluid left inside adapter body 101.

[0044] The mixing member 118 may have a mixing chamber 124 configured to receive the mixing pin 160. The mixing pin 160 may be removably insertable into the mixing chamber 124, for example, to allow interchangeability of the mixing pins 160, 260, 360, 460, 660, 760 depending on the intended use and / or substance. More specifically, the geometry of the mixing pins 160, 260, 360, 460, 660, 760 may affect the properties of the pharmaceutical conjugate such that the adapter 100 is configured to produce different types of LNPs based on the intended use. The adapter body 101 may have an opening 126 aligned longitudinally and in communication with the mixing chamber 124. The mixing pin 160 may be inserted into the mixing chamber 124 through the opening 126. The opening 126 may be located at the top of the body 101, between the first port 102 and the second port 104, and may be in communication with the top of the mixing chamber 124. As further shown, the mixing pin 160 may include a flange 162, and the body 101 may include a countersunk recess 128 configured to receive the flange 162 when the mixing pin 160 is inserted into the mixing chamber 124. To ensure that the mixing pin 160 is inserted into the mixing chamber 124 in a particular orientation, the opening 126 and / or the recess 128 may have a non-circular and / or axially asymmetric cross-section, and the flange 162 may have a non-circular and / or axially asymmetric cross-section corresponding to the opening 126 and / or the recess 128. To ensure that the mixing pin 160 is properly aligned with the branch channels 120, 122, the recess 128 and the flange 162 may each be non-circular but bilaterally symmetric, e.g., substantially D-shaped in cross-section. The cross-sections of the recess 128 and the flange 162 may similarly be rectangular and / or elliptical. In some embodiments (as shown in Figures 2-4), the recess 128 may be in a tubular member 129 that extends upward from the mixing member 118, between the branch members 114, 116. As shown in Figure 4, the tubular member 129 may have a width or diameter greater than the width of the mixing member 118 to receive the flange 162. In some embodiments (as shown in Figure 8), the tubular member 129 may be omitted.The mixing pin 160 may be releasably secured within the mixing chamber 124 through a press fit of the flange 162 inside the recess 128. However, the mixing pin 160 may additionally or alternatively be releasably secured within the mixing chamber 124 by a snap engagement, clip, threaded connection, and / or bayonet connection. The mixing chamber 124 may have a substantially uniform cross-section beneath the branch channels 120, 122 that at least partially define at least one microfluidic flow path and / or microfluidic channel for the flow of the first and second substances.

[0045] The mixing pin 160 may have a sealing portion 164 that extends below the flange 162. The sealing portion 164 may have a width or diameter that is smaller than the width or diameter of the flange 162 and may be received below the flange 162 in the top of the mixing chamber 124. The sealing portion 164 may have a shape configured to seal the top of the mixing chamber 124. The sealing portion 164 may be cylindrical.

[0046] At least one flow path may be defined at least partially around the mixing pin 160 for mixing a first stream of a first material and a second stream of a second material through turbulent flow. The mixing pin 160 may include opposite sides, each facing one of the branch channels 120, 122. Each of the sides may be configured to split the flow from one of the branch channels 120, 122 into two separate flow streams. For example, the side of the mixing pin 160 facing the first branch channel 120 may have first and second surfaces 166 that meet at a first edge 168. The side of the mixing pin 160 facing the second branch channel 122 may have first and second surfaces 170 that meet at a second edge 172. The mixing pin 160 may be inserted into the adapter body 101 such that the first edge 168 and the second edge 172 are aligned with the transverse axis of the body 101. The first edge 168 may be aligned with the first branch channel 120, and the second edge 172 may be aligned with the second branch channel 122, and the edges 168, 172 may be adapted to guide or divert fluid flow from the branch channels 120, 122 around the mixing pin 160. The first edge 168 may divide the fluid flow from the first branch channel 120 into a first fluid path along a first one of the surfaces 166 and a second fluid path along a second one of the surfaces 166. Similarly, the second edge 172 may divide the fluid flow from the second branch channel 122 into a first fluid path along a first one of the surfaces 170 and a second fluid path along a second one of the surfaces 170. Each of the surfaces 166, 170 may be substantially flat, and the edges 168, 172 may be substantially sharp, blunt, or slightly rounded to reduce turbulence of the pre-mixed flows from the branch channels 120, 122. The mixing pin 160 may have one or more side surfaces 174 on each side of the mixing pin 160. Each of the side surfaces 174 may extend between a pair of the surfaces 166, 170. As shown in FIG. 4 , the side surfaces 174 may define a radial width smaller than that of the seal portion 164 to define a space inside the mixing chamber 124 for initial mixing of the flows of the first and second materials.The lateral surfaces 174 may further allow for longitudinal movement of fluid flow.

[0047] At least one flow path may be serpentine. For example, the at least one flow path may be at least partially defined by at least one helical channel 176, 178. The at least one helical channel 176, 178 may include a first helical channel 176 and a second helical channel 178. The first helical channel 176 and the second helical channel 178 may have different orientations and / or extend in opposite helical directions. At least one side surface 174 may extend from the surface / edge 166 below 172 and be in fluid communication with at least one of the helical channels 176, 178. As shown, the mixing pin 160 may include a pair of side surfaces 174, each in fluid communication with at least one of the first helical channel 176 and the second helical channel 178. However, in some embodiments, the mixing pin 160 may have a single side surface that is in fluid communication with both the first spiral channel 176 and the second spiral channel 178. The first and second spiral channels 176, 178 may intersect at multiple intersections 180 on opposing side surfaces of the mixing pin 160 along the longitudinal length of the mixing pin 160 to generate turbulent flow and mix the first and second materials. The first and second spiral channels 176, 178 may form a double helix. As the components collectively pass through the mixing chamber 124, turbulence is induced by the serpentine flow path around the mixing pin 160, thus transitioning the components from an unmixed state to a mixed state. As the fluid passes through the intersection 180, a local change in fluid flow direction is induced. Thus, the degree of turbulence (i.e., the Reynolds number) of the fluid flow increases as the fluid passes through the intersection 180. As the degree of turbulence increases, the degree of parallelism of the components of the fluid stream decreases, thus increasing the degree of mixing. Consequently, increasing the turbulence of the fluid within the mixing chamber can promote mixing of the components of the fluid. A pharmaceutical complex is thereby formed by the first and second components collectively passing through the mixing chamber 124. The pharmaceutical complex may exit the first and second spiral channels 176, 178 of the mixing pin 160, pass through the bottom of the adapter body 101, and then exit the adapter 100 through the tip channel 125 and into the receiving vessel 60.

[0048] Thus, the geometry of the first and second helical channels 176, 178 may be selected depending on the degree of turbulence needed to achieve the amount of mixing required for any given application of the adapter 100. For example, the pitch of the first and second helical channels 176, 178 may be designed for the desired turbulence. The first and second helical channels 176, 178 may have a smaller pitch and a larger number of intersections 180 for applications where a higher degree of turbulence is required to achieve the desired mixing (e.g., lipid nanostructure formation). However, as the number of intersections 180 increases, the resistance to fluid flow through the mixing chamber 124 also increases. On the other hand, the first and second helical channels 176, 178 may have a steeper pitch and fewer intersections 180 for applications where a lower degree of turbulence is required to achieve the desired mixing, or where lower flow volatility is required to maintain certain physical properties of the components during mixing. Thus, the pitch of the first and second helical channels 176, 178 may be selected to provide a desired degree of mixing while avoiding impractically high resistance for use with manually driven syringes. With this in mind, in one implementation, the inventors have found that an arrangement of 8 to 40 intersections 180, e.g., 8 to 12 intersections 180, is particularly useful for producing mRNA-LNP or RNA-LNP compositions. However, as the reader will understand, a different number of turns may be used in different situations.

[0049] The first and second spiral channels 176, 178 may be formed from grooves or depressions in the outer surface of the mixing pin 160. The first and second spiral channels 176, 178 may be microfluidic pathways to reduce the effect of body forces on fluid flow. The first and second spiral channels 176, 178 may have a width ranging from about 100 μm to about 1000 μm and / or a depth ranging from about 100 μm to about 1000 μm. However, the width and / or depth is preferably about 400 μm to about 1000 μm to facilitate manufacturing. In a preferred embodiment, the inventors have found consistent and rapid formation of pharmaceutical complexes (e.g., mRNA-LNP) when the width and / or depth of the first and second spiral channels 176, 178 are about 400 μm to about 600 μm. Smaller channels may be used to increase the velocity of fluid flow through the mixing chamber 124, which may further improve mixing. At these scales, the effect of the tortuous flow path on fluid turbulence may be amplified, resulting in increased mixing, compared to fluid paths with larger channels. The higher the fluid velocity through the mixing chamber 124, the greater the amount of turbulence induced, and therefore the greater the degree of turbulence induced throughout the mixing chamber 124. However, smaller channels increase pressure and resistance. Three-dimensional flow paths or channels as disclosed increase the overall flow volume through the adapter 100, reducing the required pressure applied by the containers 20, 40, thereby enabling optimal formation of pharmaceutical complexes.

[0050] The adapter body 101 may have a third port or connector member 130 at the bottom of the mixing member 118. The third connector member 130 may be a male luer connector including a tip 132 and a sleeve 134 configured to be attached to the receiving container 60 via the vial adapter 70. The tip 132 may have a tip channel 125 that communicates with a second portion or bottom of the mixing chamber 124. The tip 132 may be received within the connector 74 of the vial adapter 70, and the sleeve 134 may be threadably connected to an outer surface of the connector 74 with a luer connection. The third connector member 130 may be a male luer connector configured to connect to the female luer connector of the vial adapter 70. However, the connector member 130 may additionally or alternatively connect to the receiving container 60 with other types of connections, such as a snap fit and / or a press fit. The tip channel 125 may provide passage for the mixed composition from the mixing chamber 124 to the receiving container 60.

[0051] The adapter body 101 may be formed from a polymer, metal, and / or glass. In a preferred embodiment, the adapter body 101 may be formed (e.g., through injection molding or three-dimensional printing) as a single, integral part including the first port 102, the second port 104, the mixing member 118, the tubular member 129, and / or the connector member 130. Alternatively, the adapter body 101 may be formed from two parts (e.g., halves) secured or fused together, each of which may be metal, polymer, or glass. A low surface energy material may be used for at least a portion of the adapter body 101 to increase the ease with which the mixing fluids flow through the adapter 100. For example, the adapter body 101 and / or the mixing pin 160 may be formed from or coated with a low surface energy material such as ethylene tetrafluoroethylene (ETFE). Other low surface energy materials, such as fluoropolymer materials other than ETFE, may also be used. Alternatively, at least one passageway may be treated to reduce its surface energy. Forming the sides of the channel of a low surface energy material may reduce loss of components across the mixing chamber 124 during use, and therefore may allow the adapter 100 to operate more efficiently. The low surface energy material may provide additional benefits in some embodiments, but is an optional feature of the present disclosure.

[0052] 8 and 9 show a second embodiment of the mixing pin 260 in the adapter 100 of FIGS. 1-4. The disclosure of the mixing pin 160 is expressly incorporated herein unless otherwise indicated. As discussed further herein, the mixing pin 260 may be inserted into the mixing chamber 124 of the adapter 100 with the flange 262 received in the recess 128 and the seal 264 sealing the top of the mixing chamber 124. A first substance may be introduced into the first port 102, and a second substance may be introduced into the second port 104. The first substance may pass through the first connector channel 110 and the first branch channel 120 and enter the mixing chamber 124. The second substance may pass through the second connector channel 114 and the second branch channel 122 and enter the mixing chamber 124. The mixing pin 260 may include a plurality of radial protrusions 282 positioned radially between the mixing pin 260 and the inner wall of the adapter body 101, defining a first channel 284 longitudinally therebetween. The protrusions 282 may extend at least partially around the circumference of the mixing pin 260, forming at least one microfluidic flow path. For example, the protrusions 282 may extend uniformly around the entire circumference of the mixing pin 260, forming a constriction within the mixing chamber 124. In some embodiments, the protrusions 282 may have a substantially uniform outer dimension (e.g., circumference) that is slightly smaller than the inner dimension (e.g., circumference) of the inner wall of the adapter body 101. The second channel 286 may be defined by a gap between the outer dimension of the protrusions 282 and the inner wall of the adapter body 101, inducing turbulence for suitable mixing of the materials. The second channel 286 may be smaller than the first channel 284, forming the constriction. In some embodiments, the mixing pin 260 may create a microfluidic pathway having at least two alternating dimensions, including the first channel 284 and the second channel 286, such that the cross-sectional area of ​​the pathway varies along its length. By including at least two alternating internal dimensions, the fluid flowing through the mixing chamber 124 undergoes repeated acceleration and deceleration, thus inducing additional turbulence in the mixing chamber 124.In a preferred embodiment, the first channel 284 may have a diameter or width of about 250 μm to about 1000 μm, and the second channel 286 may have a width or diameter of about 50 μm to about 200 μm.

[0053] 10-12 illustrate a third embodiment of a mixing pin 360 of the system 10 of FIGS. 1-4. The disclosure of mixing pins 160 and / or 260 is expressly incorporated herein unless otherwise indicated. As discussed further herein, the mixing pin 360 may be inserted into the mixing chamber 124 of the adapter 100 with the flange 362 received in the recess 128 and the seal 364 sealing the top of the mixing chamber 124. A first substance may be introduced into the first port 102, and a second substance may be introduced into the second port 104. The first substance may pass through the first connector channel 110 and the first branch channel 120 and enter the mixing chamber 124. The second substance may pass through the second connector channel 114 and the second branch channel 122 and enter the mixing chamber 124.

[0054] The at least one microfluidic flow path may be serpentine. The at least one flow path may be longitudinal through, along, and / or circumferentially around the mixing pin 360. As further shown, the mixing pin 360 may include at least one first protrusion 382 and at least one second protrusion 383 disposed along the longitudinal length of the mixing pin 360. The at least one first protrusion 382 and at least one second protrusion 383 may be longitudinally spaced apart by a first channel 384 defined by a reduced width or diameter portion of the mixing pin 360. Each of the at least one first protrusion 382 and at least one second protrusion 383 may be laterally symmetrical. For example, each of the protrusions 382, ​​383 may include at least one first surface 388 configured to form a seal against the inner surface of the adapter body 101 when inserted into the mixing chamber 124, and at least one second surface 389 configured to form a second channel 386 formed by a radial gap against the inner surface of the mixing element 118, allowing longitudinal passage of flow through the mixing chamber 124 and generating mixing between opposing circumferential flows around the first channel 384 of the mixing pin 360. The at least one first surface 388 may be curved to approximate the inner wall of the adapter body 101, and the at least one second surface 389 may be flat to form the second channel 386 against the inner wall of the adapter body 101. For example, the protrusions 382, ​​383 may have a substantially rectangular geometry having a first width defining a pair of first rounded sides 388 and a second width defining a pair of second straight sides 389, the first width being greater than the second width. However, it is also contemplated that one or more or all of the protrusions 382, ​​383 may define only a single second surface 389, and thus only a single second channel 386, with the remaining circumference being defined by the curved first surface 388. Thus, the second channel 386 may be smaller than the first channel 384. For example, the second channel 386 may be about 250 μm to about 1000 μm, and the groove 384 may be about 50 μm to 200 μm.

[0055] The first channel 384 and the second channel 386 may collectively define at least one channel along the longitudinal length of the mixing pin 360 to provide at least one flow path. As further shown in FIGS. 10 and 11 , the first and second protrusions 382, ​​383 may be radially offset such that the second channel 386 is not aligned longitudinally along the mixing chamber. The first and second protrusions 382, ​​383 may alternate. For example, in a preferred embodiment (as shown in FIG. 10 ), longitudinally adjacent first and second protrusions 382, ​​383 may be offset by 90 degrees and arranged longitudinally in the following order: one first protrusion 382 followed by one second protrusion 383, another first protrusion 382 followed by another second protrusion 383, and so on. The non-alignment of the second channels 386 may enhance mixing of the materials by requiring the materials to move circumferentially around the mixing pin 360 to pass through at least one first surface 388, with opposing fluid flows converging along the mixing pin 360 at the second channels 386. However, it is contemplated that other arrangements of additional protrusions may be provided.

[0056] As further shown, the mixing pin 360 may define a closed channel or lumen 390 through the top below the seal 364 configured to receive fluid streams, perform initial mixing, and / or introduce fluid streams into at least one path of the mixing pin 360 and the mixing chamber 124. The lumen 390 may have a pair of inlet openings 391 each communicating with one of the branch channels 120, 122. The lumen 390 may have a first segment 390a extending transversely to the longitudinal axis of the mixing pin 360, where opposing fluid streams from the branch channels 120, 122 converge for initial mixing. The lumen 390 may have a second segment 390b extending along the longitudinal axis of the mixing pin. Fluid may travel through the second segment 390b, through a second transverse segment (not shown), and exit through one or more outlet openings 392 to enter at least one pathway defined by the first and second protrusions 382, ​​383. The lumen 390 may have a pair of outlet openings 392, each longitudinally aligned with one of the first sides 388 and requiring circumferential fluid flow to pass through one of the first channels 384. The inlet openings 391 may each extend through an edge 368 formed by adjacent surfaces 366. The first edge 368 may be aligned with the first branch channel 120, and the second edge (not shown) may be aligned with the second branch channel 122. The surface 366 may extend to an outer diameter of the mixing pin 360, where it contacts the inner surface of the mixing chamber 124. Second sealing portion 365 may be below surface 366 and may have a shape configured to seal around mixing pin 360 and force it through inlet opening 391. Second sealing portion 365 may be cylindrical.

[0057] FIG. 13 illustrates a fourth embodiment of a mixing pin 460 of the system 10 of FIGS. 1-4. The disclosure of mixing pins 160, 260, and / or 360 is expressly incorporated herein unless otherwise indicated. As discussed further herein, the mixing pin 460 may be inserted into the mixing chamber 124 of the adapter 100 with the flange 462 received in the recess 128 and the seal 464 sealing the top of the mixing chamber 124. A first substance may be introduced into the first port 102, and a second substance may be introduced into the second port 104. The first substance may pass through the first connector channel 110 and the first branch channel 120 and enter the mixing chamber 124. The second substance may pass through the second connector channel 114 and the second branch channel 122 and enter the mixing chamber 124.

[0058] At least one passage may be defined at least partially around the mixing pin 460 for mixing a first stream of a first material and a second stream of a second material through turbulent flow. The mixing pin 460 may include a side facing one of the branch channels 120, 122 configured to split the flow from at least one of the branch channels 120, 122 into two separate flow streams. For example, the side of the mixing pin 460 facing the first branch channel 120 may have first and second surfaces 466 that meet at a first edge 468. Additionally or alternatively, the side of the mixing pin 460 facing the second branch channel 122 may have first and second surfaces 470 that meet at a second edge (not shown). The first edge 468 and the second edge may be aligned with a transverse axis of the mixing pin 460 and / or a transverse axis of the body 101. The first edge 468 may be aligned with the first branch channel 120, and the second edge 172 may be aligned with the second branch channel 122, and the edges 468 may be adapted to guide or divert fluid flow from the branch channels 120, 122 around the mixing pin 460. The first edge 468 may divide the fluid flow from the first branch channel 120 into a first fluid path along a first one of the surfaces 466 and a second fluid path along a second one of the surfaces 466. Similarly, the second edge may divide the fluid flow from the second branch channel 122 into a first fluid path along a first one of the surfaces 470 and a second fluid path along a second one of the surfaces 470. Each of the surfaces 166, 170 may be substantially flat, and the edges 468 may be substantially sharp, blunt, or slightly rounded to reduce turbulence in the flow from the branch channels 120, 122. The mixing pin 460 may have one or more side surfaces 474 on each side of the mixing pin 460. Each of the side surfaces 474 may extend between a pair of surfaces 466, 470. As shown, the side surfaces 474 may define a radial width that is smaller than that of the seal portion 464 to define a space inside the mixing chamber 124 for initial mixing of the streams of the first and second materials.

[0059] The at least one pathway of the mixing pin 460 may include a first pathway along a first side of the mixing pin 460 and a second pathway along a second side of the mixing pin 460. A first side surface of the side surface 474 may extend downwardly and define an inner surface of the first pathway, and a second side surface of the side surface 474 may extend downwardly and define an inner surface of the second pathway. The side surfaces 474 may be on opposite sides of the mixing pin 460 and may be separated by first and second convex sealing surfaces 465. The sealing surfaces 465 may abut an inner surface of the mixing chamber 124 to prevent passage of fluid between the first pathway and the second pathway. The sealing surfaces 465 may extend from an edge 468 at an upper portion of the mixing chamber 124 to a lower portion of the mixing chamber 124 to separate the first and second fluid pathways into two separate fluid pathways. The first fluid pathway may include a plurality of first protrusions 482 extending or protruding laterally from the first lateral surface 474. The plurality of first protrusions 482 may form an obstacle and flow constriction in the fluid pathway that creates turbulence to generate mixing within the first fluid pathway. Similarly, the second fluid pathway may include a plurality of second protrusions 484 extending or protruding laterally from the second lateral surface 474. The plurality of second protrusions 484 may form an obstacle and flow constriction in the fluid pathway that creates turbulence to generate mixing within the second fluid pathway. For example, the protrusions 482, 484 may divide each of the fluid pathways and / or form narrow gaps therebetween to constrict the fluid flow and induce localized accelerations, intersections, and / or vortices in the fluid flow. Each of the protrusions 482, 484 may have an angled or V-shaped proximal portion that divides the fluid flow. One of the split streams may follow a path formed between the rounded distal portion of a protrusion 482, 484 and the concave portion of a distally adjacent protrusion 482, 484 and collide with the other split stream, creating vortices and turbulence. The protrusions 482, 484 on each side surface 474 may form a modified Tesla valve, providing turbulence and improving mixing. The first and second paths may meet at the bottom end of the mixing chamber 124 and / or at the tip channel 125 and be routed into the receiving vessel 60.

[0060] FIG. 14 illustrates a second embodiment of an adapter 500 for the system 10 of FIGS. 1-4. The adapter 500 may include a body 501 having a first port 502 configured to attach to the syringe connector 26 of the first container 20 and a second port 504 configured to attach to the syringe connector 46 of the second container 40. The adapter body 501 may further include a first branch member 514, a second branch member 516, and a mixing member 518. The first port 502 may extend from the first branch member 514, which may connect the first port 502 to the mixing member 518. The second port 504 may extend from the second branch member 516, which may connect the second port 504 to the mixing member 518. The first branch member 514 may have a first branch channel 520 configured to receive a fluid flow from the first syringe 20 through the first port 502, and the second branch member 516 may have a second branch channel 522 configured to receive a fluid flow from the second syringe 40 through the second port 504. The first branch channel 520 and the second branch channel 522 may be angled relative to a longitudinal axis of the adapter body 501 and / or the mixing pin 160 to converge the fluid flow from the first syringe 20 and the second syringe 40 into the mixing member 518. The angle of at least one of the first branch channel 520 and the second branch channel 522 may be greater than 90 degrees relative to the mixing chamber 524, the adapter body 501, and / or the mixing pin 160. As further shown, the angle of at least one of the first branch channel 520 and the second branch channel 522 may be from about 120 degrees to about 160 degrees relative to the mixing chamber 524, the adapter body 501, and / or the mixing pin 160. However, as discussed herein, other configurations may be employed for other applications. The first branch channel 520 and the second branch channel 522 may be aligned with a midplane of the adapter body 501 such that the first branch channel 520 and the second branch channel 522 direct the first and second substances toward opposite sides of the mixing pin 160. The first branch channel 520 and the second branch channel 522 may form a Y-shape with the mixing element 518.The disclosure of adapter 100 is expressly incorporated herein in its entirety unless otherwise indicated. Although shown with mixing pin 160, adapter 500 may include any of the other disclosed mixing pins.

[0061] As further shown, the adapter body 501 may be formed separately from at least one or all of the ports 102, 104, 130. For example, the adapter body 501 may be formed through injection molding or three-dimensional printing, and the ports 102, 104, 130 may be formed separately and engaged to the adapter body 501 through a press fit, threaded engagement, and / or snap engagement. The adapter 500 may further include a retainer 594 configured to be applied to the adapter body 501 to secure the mixing pin 160 within the mixing chamber 124. As shown, the retainer 594 may include a plate secured to the adapter body 501 using one or more fasteners 195, which may be threaded. However, the retainer 594 may additionally or alternatively include other structures, such as a clip, a sliding or pivoting door, or the like. The adapter 500 may further include a sealing member 595 configured to seal the proximal end of the mixing chamber 524. The sealing member 595 may be a separate component from the mixing pin 160. For example, the sealing member 595 may be an elastomeric O-ring disposed around a proximal portion of the mixing pin 160, such as the sealing portion 164. The adapter 500 may include a plug 540 inserted into one or both of the branch members 514, 516 to seal the outer ends of the respective branch channels 520, 522. The plug 540 may have an interface for receiving a distal effector of a tool for adjustment and / or removal. For example, as shown, the plug 540 may include one or more slots that may engage with a flat-head, Phillips®, Torx®, and / or hex-head effector of a tool or screwdriver.

[0062] 15-24 illustrate a third embodiment of an adapter 600 for the system 10 of FIGS. 1-4. As shown, the adapter 600 may include a body 601 having a first port or connector member 602 configured to attach to the syringe connector 26 of the first container 20 and a second port or connector member 604 configured to attach to the syringe connector 46 of the second container 40. The adapter 600 may include a mixing member 618 that receives a mixing pin 660. The disclosures of the adapters 100, 500 and / or mixing pins 160, 260, 360, 460 are expressly incorporated herein in their entirety unless otherwise indicated.

[0063] The first port 602 may be configured to be received by the first syringe connector 26 and may have external threads 606 configured to mate with the internal threads of the first syringe connector 26. The second connector 604 may be configured to be received by the second connector 46 and may have external threads 608 around the second tubular member 608 configured to mate with the internal threads of the second syringe connector 46. For example, the first port 602 may be a female Luer connector and the connector 26 of the first syringe 20 may be a male Luer connector. However, the ports 602, 604 may additionally or alternatively connect to the containers 20, 40 with other types of connections, such as a snap fit, a friction fit, and / or a press fit. The adapter 600 may be configured to connect to any number of first and second containers 20, 40, and the adapter 100 may therefore have an equal number of ports 602, 604 for connecting to each of the containers 20, 40. Additionally, one or more of the ports 602, 604 may include a one-way valve (not shown) to allow fluid flow from the container 20, 40 to the adapter 600 and to restrict or substantially prevent fluid flow from the adapter 600 back to the respective container 20, 40.

[0064] The first port 602 may define a first connector channel 610, and the second connector member 604 may define a second connector channel 612. The first connector channel 610 may be configured to receive a tip of the first syringe 20 to fluidly connect a chamber of the first syringe body 22 to the first connector channel 610, and the second connector channel 612 may be configured to receive a tip of the second syringe 40 to fluidly connect a chamber of the second syringe body 42 to the second connector channel 612. The first port 602 and the second port 604 may extend substantially parallel to one another to facilitate coordinated actuation of the syringes 20, 40. In some embodiments, the first port 602 and the second port 604 may extend substantially parallel to a longitudinal axis of the adapter body 601, as shown in FIG. 15 .

[0065] The adapter body 601 may further include a first branch member 614 and a second branch member 616. The first port 602 may extend from the first branch member 614, which may connect the first port 602 to a mixing member 618. The second port 604 may extend from the second branch member 616, which may connect the second port 604 to the mixing member 618. The first branch member 614 may have a first branch channel 620 configured to receive a fluid flow from the first syringe 20 through the first connector channel 610, and the second branch member 616 may have a second branch channel 622 configured to receive a fluid flow from the second syringe 40 through the second connector channel 612. The first and second branch channels 620 and 622 may communicate with the top of the mixing element 618. The first and second branch channels 620 and 622 may be angled relative to the longitudinal axis of the adapter body 601 and / or the mixing pin 660 to converge fluid flow from the first and second syringes 20 and 40 into the mixing element 618. The angle of at least one of the first and second branch channels 620 and 622 may be greater than 90 degrees relative to the mixing element 618, the adapter body 601, and / or the mixing pin 160. As further shown, the angle of at least one of the first and second branch channels 620 and 622 may be from about 120 degrees to about 160 degrees relative to the mixing element 618, the adapter body 601, and / or the mixing pin 160. However, as discussed herein, other configurations may be used in other applications. First branch channel 620 and second branch channel 622 may be oriented offset from or misaligned with the mid-plane of adapter body 601 (as indicated by central axis A, as shown in FIG. 16 ). In some embodiments, first branch channel 620 and second branch channel 622 may have the same or substantially the same width or diameter (as shown in FIG. 15 ) for applications where the same flow rates are desired for the first and second substances.In some embodiments (as shown in FIGS. 11 and 12 ), the first branch channel 620 and the second branch channel 622 may have different widths or diameters for applications in which different flow rates are desired for the first and second substances. For example, (as further shown in FIGS. 11 and 12 ), the first branch channel 620 may have a larger width or diameter than the second branch channel 622. In some embodiments (as shown in FIGS. 2 , 3 , and 8 ), at least one or both of the first branch channel 620 and the second branch channel 622 may be formed with open outer ends, and the adapter 600 may include plugs 640 inserted into one or both of the branch members 614, 616 to seal the outer ends of the respective branch channels 620, 622. In some embodiments (as shown in FIG. 11 ), at least one or both of the first branch channel 620 and the second branch channel 622 may have closed outer ends formed by the adapter body 601.

[0066] The plugs 640a, b may be removable, replaceable, interchangeable, and / or adjustable. The plugs 640a, b may facilitate manufacturing of the adapter body 601 by allowing the branch channels 620, 622 to be formed in the adapter body 601 through injection molding by inserting a straight pin (not shown). As expressly incorporated herein, the pin may be removed from the injector body 601 after being formed through the opening, and the plug 640 may be inserted to seal the opening of the branch channels 620, 622, as discussed above with reference to the plug 140. Replacing and / or adjusting the plug 640a may change the inlet volume of one or both of the first connector channel 610 and / or the second connector channel 612. For example, the plugs 640a, b may be replaced with similar plugs 640a, b having different lengths and / or geometries to at least partially occlude one or both of the first connector channel 610 and / or the second connector channel 612. In some embodiments, the plug 640 may be adjusted by pushing or translating it into the branch channels 620, 622 to at least partially block the first connector channel 610 and / or the second connector channel 612. In some embodiments, the plug 640 may have an adjustable length, such as being telescoping, to at least partially block the first connector channel 610 and / or the second connector channel 612. Varying the inlet volume using the plugs 640a, b may adjust the amount of fluid mixing and / or reduce the dead volume of fluid left inside the adapter body 601. As shown in FIGS. 23-24 , the plugs 640a, b may include an end cap 641 and a pin 642. The end cap 641 may be configured to attach to the branch channels 620, 622 to secure the plugs 640a, b. The pin 642 may be configured to extend into at least one of the branch channels 620, 622.As discussed further with respect to plug 140 (which discussion is expressly incorporated herein), pin 642 may be configured to occupy space within branch channels 620, 622 to reduce negative space. Plugs 640 a, b may further include grooves 643 configured to receive a sealing member (not shown). Plugs 640 a, b may be the same or different for each branch channel 620, 622. For example, plug 640 a, b for one of branch channels 620, 622 may include pin 642, while plug 640 a, b for another of branch channels 620, 622 may not include pin 642.

[0067] The mixing member 618 may have a mixing chamber 624 configured to receive a mixing pin 660. The mixing pin 660 may be removably inserted into the mixing chamber 624, for example, to allow for interchangeability of the mixing pin depending on the intended application and / or substance. The body 601 may have an opening 626 aligned longitudinally and in communication with the mixing chamber 624. The mixing pin 660 may be inserted into the mixing chamber 624 through the opening 626. The opening 626 may be in an upper portion of the body 601, between the first port 602 and the second port 604, and may be in communication with an upper portion of the mixing chamber 624. As further shown, the mixing pin 660 may include an end cap 662, and the body 601 may include a rim 628 configured to engage the end cap 662 when the mixing pin 660 is inserted into the mixing chamber 624. The rim 628 may be on a tubular member 629 extending upward from the mixing member 618, between the branch members 614, 616. The end cap 662 may have a downwardly extending sleeve portion 663 configured to receive or surround the rim 628 of the body 601. The sleeve portion 663 may include one or more flexible protrusions 665 configured to flex radially outward when the rim 628 is received within the sleeve portion 663 and to releasably engage the rim 628 to secure the mixing pin 660 within the mixing chamber 624 via a press fit and / or snap fit. The one or more flexible protrusions 665 may be cantilevered from the top of the end cap 662. However, the mixing pin 660 may additionally or alternatively be releasably secured within the mixing chamber 624 via other types of snap fit, press fit, threaded connection, and / or bayonet connection.

[0068] The mixing pin 660 may have a relatively large diameter, such as greater than 8 mm. The diameter of the mixing pin 660 may provide design flexibility for the configuration of the first branch channel 620 and / or the second branch channel 622. As discussed above, the first branch channel 620 and / or the second branch channel 622 may be positioned or oriented so that they are offset or misaligned from the transverse axis A (representing the midplane) of the adapter body 601 and / or the mixing pin 660 when inserted into the mixing chamber 624. This offset feature of the first branch channel 620 and / or the second branch channel 622 may generate angular momentum and potential vortices in the fluid flow around the mixing pin 660. In some embodiments (as shown in FIGS. 15 through 20 ), the first branch channel 620 and the second branch channel 622 may be positioned on the same side of the transverse axis A such that the first and second substances are introduced into the mixing chamber 624 in opposite directions and immediately collide. In some embodiments (as shown in FIGS. 25-26 ), the first branch channel 620 and the second branch channel 622 may be located on opposite sides of the horizontal axis A of the body 601 such that the first substance and the second substance may flow in the same circumferential direction when introduced into the mixing chamber 624. In some embodiments (not shown), one of the branch channels 620, 622 may be oriented in alignment with the horizontal axis A, and the other of the branch channels 620, 622 may be oriented offset or not aligned with the horizontal axis A. In some embodiments (not shown), the branch channels 620, 622 may be connected to the adapter body 601 at different longitudinal positions. For example, the first branch channel 620 may be connected to the adapter body 601 at a position above or below the second branch channel 622.

[0069] At least one passage may be defined at least partially around the mixing pin 660 for mixing the first stream of the first material and the second stream of the second material through turbulent flow. The mixing pin 660 may have a sealing portion 664 extending below the end cap 662. The sealing portion 664 may have a shape configured to seal the top of the mixing chamber 624. The sealing portion 664 may have a width or diameter smaller than the width or diameter of the end cap 662 and may be received below the end cap 662 in the top of the mixing chamber 624. The sealing portion 664 may be cylindrical. The mixing pin 660 may have a channel portion 667 below the sealing portion 664, the channel portion 667 having a width or diameter smaller than the sealing portion 664. The channel 667 may additionally or alternatively receive a sealing member 695, which is a separate component, such as an elastomeric O-ring. The mixing pin 660 may further include a threaded outer portion 675 defining at least one helical channel 676. The threaded outer portion 675 may have a width or diameter greater than the channel 667, and the threaded outer portion 675 may extend the majority of the length of the mixing pin 660 (e.g., more than half its length and / or more than two-thirds its length).

[0070] Thus, the geometry of the at least one helical channel 676 may be selected depending on the degree of turbulence needed to achieve the amount of mixing required for any given application of the adapter 600. For example, the pitch of the at least one helical channel 676 may be designed for the desired turbulence. The at least one helical channel 676 may have a low pitch for applications where a higher degree of turbulence is needed to achieve the desired mixing (e.g., lipid nanostructure formation). On the other hand, the at least one helical channel 676 may have a steeper pitch for applications where a lower degree of turbulence is needed to achieve the desired mixing, or where lower flow volatility is needed to maintain certain physical properties of the components during mixing. Thus, the pitch of the at least one helical channel 676 may be selected to provide the desired degree of mixing while avoiding impractically high resistance for use with manually driven syringes. In some embodiments, the at least one helical channel 676 may comprise a double helix, as discussed and shown with respect to the embodiments of Figures 5 through 7, which are expressly incorporated herein by reference. The at least one spiral channel 676 has a width of about 200 μm to about 1000 μm. In a preferred embodiment, the at least one spiral channel 676 may have a width of about 400 μm to about 600 μm to facilitate manufacturing and optimize mixing and hydraulics. Additionally, the bottom of the at least one spiral channel 676 may be spaced apart from the inner surface of the adapter body 601 by about 200 μm to about 1000 μm. In a preferred embodiment, the bottom of the at least one spiral channel 676 may be spaced apart from the inner surface of the adapter body 601 by about 400 μm to about 600 μm.

[0071] The body 601 may have a third port or connector member 630 at the bottom of the mixing member 618. The third port 630 may be a male luer connector including a tip 632 and a sleeve 634 configured to be attached to the receiving container 60 via the vial adapter 70. The tip 632 may have a tip channel 625 that communicates with a second portion or bottom of the mixing chamber 624. The tip 632 may be received in the connector 74 of the vial adapter 70, and the sleeve 634 may be threadably connected to the outer surface of the connector 74 with a luer connection. The third port 630 may be a male luer connector configured to connect to the female luer connector of the vial adapter 70. However, the third port 630 may additionally or alternatively connect to the receiving container 60 using other types of connections, such as a snap fit and / or a press fit. The tip lumen 625 may provide for passage of the mixed composition from the mixing chamber 624 to the receiving container 60.

[0072] 25-28 illustrate a fourth embodiment of an adapter 700 for the system 10 of Figures 1-4. As shown, the adapter 700 may include a body 701 having a first port or connector member 702 configured to attach to the syringe connector 26 of the first container 20 and a second port or connector member 704 configured to attach to the syringe connector 46 of the second container 40. The disclosures of the adapters 100, 500, 600 and / or mixing pins 160, 260, 360, 460, 660 are expressly incorporated herein in their entirety unless otherwise indicated.

[0073] As shown, the adapter body 701 may further include a first branch member 714, a second branch member 716, and a mixing member 718. The first port 702 may extend from the first branch member 714, which may connect the first port 702 to the mixing member 718. The second port 704 may extend from the second branch member 716, which may connect the second port 704 to the mixing member 718. The first branch member 714 may have a first branch channel 720 configured to receive fluid flow from the first syringe 20 through the first connector channel 710, and the second branch member 716 may have a second branch channel 722 configured to receive fluid flow from the second syringe 40 through the second connector channel 712. The first and second branch channels 720 and 722 may communicate with the top of the mixing element 718. The first and second branch channels 720 and 722 may be angled relative to the longitudinal axis of the adapter body 701 and / or the mixing pin 760 to converge fluid flow from the first and second syringes 20 and 40 into the mixing element 718. As discussed above with reference to the adapter 600, the first and second branch channels 720 and 722 may be oriented offset from or misaligned with the midplane of the adapter body 701 (as shown by axis A in FIG. 26 ). In some embodiments, the first and second branch channels 720 and 722 may have the same or substantially the same width or diameter (as shown in FIG. 15 ) for applications in which the same flow rates are desired for the first and second substances. In some embodiments (as shown in FIGS. 11 and 12), the first branch channel 720 and the second branch channel 722 may have different widths or diameters for applications where different flow rates are desired for the first and second substances. For example, (as further shown in FIGS. 11 and 12), the first branch channel 720 may have a larger width or diameter than the second branch channel 722.In some embodiments (as shown in FIGS. 2, 3, and 8), at least one or both of the first branch channel 720 and the second branch channel 722 may be formed with an open outer end, and the adapter 700 may include a plug 740 inserted into one or both of the branch members 714, 716 to seal the outer end of the respective branch channel 720, 722. In some embodiments (as shown in FIG. 11), at least one or both of the first branch channel 720 and the second branch channel 722 may have a closed outer end formed by the adapter body 601.

[0074] The plugs 740a, b may be removable, replaceable, interchangeable, and / or adjustable. Replacing and / or adjusting the plug 740a may change the inlet volume of one or both of the first connector channel 710 and / or the second connector channel 712. For example, the plugs 740a, b may be replaced with similar plugs 740a, b having different lengths and / or geometries to at least partially occlude one or both of the first connector channel 710 and / or the second connector channel 712. In some embodiments, the plugs 740 may be adjusted by pushing or translating them into the branch channels 720, 722 to at least partially occlude the first connector channel 710 and / or the second connector channel 712. In some embodiments, the plugs 740 may have an adjustable length, such as being telescoping, to at least partially occlude the first connector channel 710 and / or the second connector channel 712. Varying the inlet volume with plugs 740a,b may adjust the amount of fluid mixing and / or reduce the dead volume of fluid left inside adapter body 701. The description of plugs 140, 640a,b is expressly incorporated herein by reference.

[0075] As will be further discussed and shown with respect to the adapter 600, the mixing member 718 may have a mixing chamber 724 configured to receive a mixing pin 760. The mixing pin 760 may be removably inserted into the mixing chamber 724, for example, to allow for interchangeability of the mixing pin depending on the intended application and / or substance. The body 701 may have an opening aligned longitudinally and in communication with the mixing chamber 724. The mixing pin 760 may be inserted into the mixing chamber 724 through the opening 726. The opening may be in an upper portion of the body 701, between the first port 702 and the second port 704, and may be in communication with an upper portion of the mixing chamber 724. As further shown, the mixing pin 760 may include a flange cap 762, and the body 701 may include a rim configured to engage the flange cap 762 when the mixing pin 760 is inserted into the mixing chamber 724.

[0076] As discussed above, the first branch channel 720 and / or the second branch channel 722 may be oriented offset from the transverse axis A (representing the midplane) of the adapter body 701 and / or the mixing pin 760 when inserted into the mixing chamber 624. This offset feature of the first branch channel 720 and / or the second branch channel 722 may generate angular momentum and potential vortices in the fluid flow around the mixing pin 760. As shown in FIGS. 25 and 26 , the first branch channel 720 and the second branch channel 722 may be located on opposite sides of the transverse axis A of the body 701 such that the first substance and the second substance may flow in the same circumferential direction when introduced into the mixing chamber 724. At least one passage may be defined at least partially around the mixing pin 760 to mix the first stream of the first substance and the second stream of the second substance through turbulence. As shown in FIG. 25 , the mixing pin 760 may have an outer surface that may be smooth. The space between the outer surface of the mixing pin 760 and the inner surface of the adapter body 701 may be 200 μm or less.

[0077] As shown in FIG. 29, the present disclosure also includes a method 1000 of mixing at least two substances to produce a pharmaceutical conjugate via any one of the adapters 100, 500, 600, 700 and / or mixing pins 160, 260, 360, 460, 660, 760 as disclosed herein.

[0078] As shown in steps 1002 and 1004, the method may include transferring a first substance from a first storage container 80 to a first container 20 and transferring a second substance from a second storage container 82 to a second container 40. However, in some embodiments, the first container 20 and / or the second container 40 may be packaged as pre-filled containers, such as pre-filled syringes, and the storage containers 80 and 82 may be omitted from the system or kit. As shown in steps 1006 and 1008, the method may include attaching the first container 20 to a first port 102, 502, 602, 702 of the adapter 100, 500, 600, 700 and attaching the second container 40 to a second port 104, 504, 604, 704 of the adapter 100, 500, 600, 700. The method may further include attaching a receiving container 60 to a port 102, 502, 602, 702 of the adapter 100. For example, as shown, one or both of the first container 20 and the second container 40 may be a variable volume container, such as a syringe. The receiving container 60 may be a fixed volume container, such as a vial, attached to the adapter 100 using a vial adapter 70.

[0079] As shown in step 1010, the method may further include introducing the first substance and the second substance into the adapter 100, 500, 600, 700 by depressing the plunger rods 24, 44 of the first container 20 and the second container 40, respectively. The first substance may pass through the first branch channel 120, 520, 620, 720 and enter the mixing chamber 124, 524, 624, and the second substance may pass through the second branch chamber 122, 522, 622, 722 and enter the mixing chamber 124, 524, 624. In some embodiments, the first branch channel 120, 520, 620, 720 and / or the second branch channel 122, 522, 622, 722 may be aligned with the transverse axis of the adapter body 101, 501, 601, 701. In some embodiments, at least one of the first branch channel 620, 720 and / or the second branch channel 622, 722 is oriented offset from the lateral axis of the adapter body 601, 701. In some embodiments, the first branch channel 620, 720 and the second branch channel 622, 722 are oriented offset from the lateral axis of the adapter body 601, 701. In some embodiments, the first branch channel 620, 720 and the second branch channel 622, 722 are on the same side of the lateral axis of the adapter body 601, 701. In some embodiments, the first branch channel 620, 720 and the second branch channel 622, 722 are on opposite sides of the lateral axis of the adapter body 601, 701.

[0080] Thus, initially, the first and second substances are unmixed with each other at the top of the mixing chamber 124, 524, 624, 724. Then, as shown in step 1012, the first and second substances may pass longitudinally along and / or circumferentially around the mixing pin 160, 260, 360, 460, 660, 760, for example, via one or more microfluidic channels, through the mixing pin, and the first and second substances may gradually transition from an unmixed state to an increasingly mixed state. The turbulence induced in the fluid flow by the adapter's mixing chamber 124, 524, 624, 724 results in the components being mixed as desired upon reaching the bottom of the mixing chamber 124, 524, 624, 724. In step 1014, mixing the first and second substances may produce a pharmaceutical complex of the first and second substances. In some embodiments, the first substance may be an aqueous solution, the second substance may be a lipid solution, and the pharmaceutical complex may include LNPs. For example, the aqueous solution may include mRNA, and the pharmaceutical complex may include mRNA-LNPs. In step 1016, the pharmaceutical complex may be received in receiving vessel 60.

[0081] In some embodiments, the method further includes disconnecting the receiving container 60 from the connector member 130 and transferring the pharmaceutical conjugate from the receiving container 60 for dilution and / or further analysis. Alternatively, the method may further include disconnecting the receiving container 60 from the connector member 130 and using the pharmaceutical conjugate directly. In some embodiments, the first container and the second container are removably connected to the first port and the second port, respectively, and the adapter may be reused. Alternatively, the first container and the second container may be permanently connected to the first port and the second port, respectively. The adapter 100, 500, 600, 700 may be configured to enter into a locking connection with one or more containers 20, 40 to prevent reuse of the adapter 100, 500, 600, 700. In some embodiments, the mixing pin 160, 260, 360, 460, 660, 760 may be removable from the adapter body 101, 601, 701. In some embodiments, the mixing pin 160, 260, 360, 460, 660, 760 may be fixed to the adapter body 101, 601, 701.

[0082] The mixing chamber 124, 524, 624, 724 may have a circular cross-section formed by the cylindrical inner surface of the adapter body 101, 601, 701. However, it is contemplated that the mixing chamber 124, 524, 624, 724 may have other cross-sections, such as oval, square, and / or rectangular. The outer surface of the mixing pin 160, 260, 360, 460, 660, 760 may have an outer surface that corresponds to the inner surface of the mixing chamber 124, 524, 624, 724 to form a microfluidic channel or flow path as discussed herein. The mixing pin 160, 260, 360, 460, 660, 760 may be solid to facilitate manufacturing, or alternatively, may be hollow with a longitudinal lumen therethrough.

[0083] It will be understood that certain terminology is used in the foregoing description for convenience and not as a limitation. The terms "a," "an," and "the" should be read to mean "at least one" unless otherwise specified. The term "comprising" should be understood to mean "including but not limited to," such that a system or method that includes certain features or steps is not limited to only the recited features or steps, but may also include features or steps that are not recited. Similarly, terms such as "over," "under," "front," "back," "right," "left," "top," "bottom," "side," etc. are used for convenience in interpreting the drawings and should not be construed as limiting.

[0084] Those skilled in the art will also understand that modifications may be made to the exemplary embodiments described herein without departing from the invention. Structural features of the systems and devices described herein may be replaced with functionally equivalent parts. Furthermore, it will be understood that features from the embodiments may be combined with one another without departing from the disclosure.

Claims

1. An adapter, The main body is a first port configured to connect to a first container for receiving a first substance; a second port configured to connect to a second container for receiving a second substance; a third port configured to connect to a receiving vessel for outputting the mixture of the first substance and the second substance; a mixing chamber having a first portion in communication with the first port and the second port, and a second portion in communication with the third port; a body including: a mixing pin inserted into the mixing chamber, the mixing pin at least partially defining at least one microfluidic path for mixing the first substance and the second substance; Includes adapter.

2. The adapter of claim 1 , wherein the body further includes an opening in the first portion of the mixing chamber configured to receive the mixing pin.

3. The adapter of claim 1 , wherein the mixing pin is removable from the mixing chamber.

4. The adapter of claim 1 , wherein the mixing pin has a flange and the body has a recess configured to receive the flange.

5. The adapter of claim 1 , wherein the mixing pin includes a seal configured to seal the first portion of the mixing chamber.

6. The adapter of claim 1 , wherein the first port and / or the second port are substantially parallel to the mixing chamber.

7. The body includes: a first branch channel in communication with the first port and extending at a first angle relative to the mixing chamber; a second branch channel in communication with the second port and extending at a second angle relative to the mixing chamber; The adapter of claim 1 further comprising:

8. The adapter of claim 7 , wherein the first angle and / or the second angle is greater than 90 degrees.

9. 9. The adapter of claim 8, wherein the first angle and / or the second angle is between about 120 degrees and about 160 degrees.

10. 8. The adapter of claim 7, further comprising a first plug inserted into an end of the first branch channel and a second plug inserted into an end of the second branch channel.

11. 8. The adapter of claim 7, wherein the first branch channel is configured to guide the first substance toward the mixing pin and the second branch channel is configured to guide the second substance toward the mixing pin.

12. 10. The adapter of claim 1, wherein the mixing pin defines a first edge surface formed by a first surface and a second surface and a second edge formed by a third surface and a fourth surface, the first edge configured to divide the first substance and the second edge configured to divide the second substance.

13. 2. The adapter of claim 1, wherein the first port is a female luer connector, the second port is a female luer connector, and the third port is a male luer connector.

14. The adapter of claim 1 , wherein the at least one microfluidic path is serpentine.

15. The adapter of claim 1 , wherein the at least one microfluidic path is formed by at least one channel.

16. 16. The adapter of claim 15, wherein at least one channel has a dimension of about 200 μm to about 1000 μm.

17. 17. The adapter of claim 16, wherein the at least one channel has a dimension of about 400 μm to about 600 μm.

18. The adapter of claim 15 , wherein the at least one channel includes at least one spiral channel.

19. 20. The adapter of claim 18, wherein the at least one helical channel includes a first helical channel and a second helical channel, the first helical channel and the second helical channel intersecting along a longitudinal length of the mixing pin at multiple intersection points.

20. The adapter of claim 15, wherein the at least one channel extends through the mixing pin.

21. 16. The adapter of claim 15, wherein the mixing pin includes a plurality of protrusions extending at least partially around a circumference of the mixing pin and defining at least a portion of the at least one channel.

22. 22. The adapter of claim 21, wherein the plurality of protrusions extend uniformly around the circumference of the mixing pin and form at least one radial gap with an inner surface of the body, the at least one radial gap forming at least a portion of the at least one channel.

23. 22. The adapter of claim 21, wherein each of the plurality of protrusions has at least one first surface configured to form a seal against an inner surface of the body and at least one second surface that forms at least one radial gap with the inner surface of the body, the at least one radial gap forming at least a portion of the at least one channel.

24. 1. A system comprising: an aqueous solution; A lipid solution; a first container configured to receive the aqueous solution; a second container configured to receive the lipid solution; A receptor vessel; An adapter, The main body is a first port configured to connect to the first container and receive the aqueous solution; a second port configured to connect to the second container and receive the lipid solution; a third port configured to connect to the receiving vessel and output the mixture of the aqueous solution and the lipid solution; a mixing chamber extending between a first portion communicating with the first port and the second port and a second portion communicating with the third port; a body having a mixing pin configured to be inserted into the mixing chamber, the mixing pin at least partially defining at least one microfluidic path for mixing the aqueous solution and the lipid solution; an adapter including: A system including:

25. 25. The system of claim 24, further comprising a second pin configured to be inserted into the mixing chamber, the second mixing pin having a different geometry than the mixing pin.

26. 25. The system of claim 24, further comprising a first storage vessel containing the aqueous solution and a second storage vessel containing the lipid solution.

27. introducing a first substance into a first port of the adapter; introducing a second substance into a second port of the adapter; mixing the first substance and the second substance in at least a microfluidic path through, along, and / or around a mixing pin in the adapter; forming a pharmaceutical conjugate of the first substance and the second substance; A method comprising:

28. 28. The method of claim 27, wherein the first substance is an aqueous solution, the second substance is a lipid solution, and the pharmaceutical complex comprises lipid nanoparticles.

29. An adapter, The main body is a first port configured to connect to a first container for receiving a first substance; a first channel in communication with the first port; a second port configured to connect to a second container for receiving a second substance; a second channel in communication with the second port; a third port configured to connect to a receiving vessel for outputting the mixture of the first substance and the second substance; a mixing chamber having a first portion in communication with the first port and the second port, and a second portion in communication with the third port; a body including: a mixing pin inserted into the mixing chamber; At least one of the first channel and the second channel is oriented offset from a midplane of the body. adapter.

30. 30. The adapter of claim 29, wherein the body further includes an opening in the first portion of the mixing chamber configured to receive the mixing pin.

31. 30. The adapter of claim 29, wherein the mixing pin is removable from the mixing chamber.

32. 30. The adapter of claim 29, wherein the mixing pin has an end cap and the body has a rim configured to engage the end cap.

33. 30. The adapter of claim 29, further comprising a sealing member configured to seal the first portion of the mixing chamber.

34. 30. The adapter of claim 29, wherein the first port and / or the second port are substantially parallel to the mixing chamber.

35. 30. The adapter of claim 29, wherein the first channel extends at a first angle relative to the mixing chamber and the second channel extends at a second angle relative to the mixing chamber.

36. 36. The adapter of claim 35, wherein the first angle and / or the second angle is greater than 90 degrees.

37. 37. The adapter of claim 36, wherein the first angle and / or the second angle is from about 120 degrees to about 160 degrees.

38. 36. The adapter of claim 35, further comprising a first plug at an end of the first channel and a second plug at an end of the second channel.

39. 36. The adapter of claim 35, wherein the first channel and the second channel are offset from a mid-plane of the mixing pin.

40. 40. The adapter of claim 39, wherein the first channel and the second channel are on the same side of a transverse axis of the body of the adapter.

41. 40. The adapter of claim 39, wherein the first channel and the second channel are on opposite sides of a transverse axis of the body of the adapter.

42. 30. The adapter of claim 29, wherein the first channel and the second channel are connected to the body at different longitudinal positions.

43. 30. The adapter of claim 29, wherein the first port is a female luer connector, the second port is a female luer connector, and the third port is a male luer connector.

44. 30. The adapter of claim 29, wherein the mixing pin defines at least one channel.

45. 45. The adapter of claim 44, wherein at least one channel has a dimension of from about 200 μm to about 1000 μm.

46. 46. ​​The adapter of claim 45, wherein the at least one channel has a dimension of about 400 μm to about 600 μm.

47. 45. The adapter of claim 44, wherein the at least one channel includes at least one spiral channel.

48. 1. A system comprising: an aqueous solution; A lipid solution; a first container configured to receive the aqueous solution; a second container configured to receive the lipid solution; A receptor vessel; An adapter, The main body is a first port configured to connect to the first container and receive the aqueous solution; a second port configured to connect to the second container and receive the lipid solution; a third port configured to connect to the receiving vessel and output the mixture of the aqueous solution and the lipid solution; a mixing chamber extending between a first portion communicating with the first port and the second port and a second portion communicating with the third port; a body having a mixing pin configured to be inserted into the mixing chamber; an adapter including: Including, At least one of the first channel and the second channel is oriented offset from a midplane of the body. system.

49. 49. The system of claim 48, further comprising a second pin configured to be inserted into the mixing chamber, the second mixing pin having a different geometric shape than the mixing pin.

50. 49. The system of claim 48, further comprising a first storage vessel containing the aqueous solution and a second storage vessel containing the lipid solution.

51. introducing a first substance into a first port of a body of an adapter through a first channel of the body of the adapter; introducing a second substance into a second port of the adapter body through a second channel of the adapter body; mixing the first substance and the second substance around a mixing pin within the body of the adapter, wherein at least one of the first channel and the second channel is oriented offset from a midplane of the body of the adapter; forming a pharmaceutical conjugate of the first substance and the second substance; A method comprising:

52. 52. The method of claim 51, wherein the first substance is an aqueous solution, the second substance is a lipid solution, and the pharmaceutical complex comprises lipid nanoparticles.

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