Cannula Assembly

Abstract

The cannula assembly includes a cannula, a polymeric support material, and a hub. The cannula has a proximal end and a distal end. The polymeric support material substantially surrounds a portion of the cannula at or near the proximal end. The hub is configured to be attached to a syringe. The polymeric support material is disposed between the cannula and the hub. The cannula, the polymeric support material, and the hub are attached by adhesive. In one embodiment, the cannula comprises fiberglass.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Patent Application No. 63 / 180,955, filed April 28, 2021, and U.S. Provisional Patent Application No. 63 / 202,432, filed June 10, 2021, each of which is incorporated by reference in its entirety herein.

[0002] FIELD OF THEINVENTION The present invention relates to a cannula assembly, and more particularly to a cannula assembly that includes a hub for connecting to a syringe. [Background technology]

[0003] Cannulas have been used in combination with syringes to deliver or receive fluids. One problem associated with the use of cannulas and the components used with cannulas is sterilization. The components of the cannula are often individually sterilized and then assembled in a sterile surgical field before being attached to the syringe. Some cannula designs are susceptible to breakage and can have an undesirable amount of dead space where unaccounted for liquids can accumulate.

[0004] It would be desirable to have a simplified method of forming a cannula assembly that can be efficiently sterilized, and to have an efficient cannula assembly design that prevents or inhibits cannula breakage and reduces dead volume when used with a syringe. Summary of the Invention [Means for solving the problem]

[0005] The term "embodiments" and similar terms are intended to refer broadly to the subject matter of the present disclosure and all of the claims that follow. Phrases including these terms should not be understood to limit the subject matter described herein or to limit the spirit or scope of the claims that follow. The embodiments of the present disclosure covered herein are defined by the claims that follow, not this Summary. This Summary is a high-level overview of various aspects of the present disclosure and introduces some of the concepts that are further described in the Detailed Description section below. This Summary is not intended to identify key or essential features of the claimed subject matter. This Summary is also not intended to be used alone to determine the scope of the claimed subject matter. The subject matter should be understood by reference to appropriate portions of the entire specification of the present disclosure, any or all drawings, and claims.

[0006] According to one aspect of the disclosure, a cannula assembly includes a cannula, a polymeric support material, and a hub. The cannula has a proximal end and a distal end. The polymeric support material substantially surrounds a portion of the cannula at or near the proximal end. The hub is configured to be attached to a syringe. The polymeric support material is disposed between the cannula and the hub. The cannula, polymeric support material, and hub are attached by adhesive.

[0007] According to one configuration of the above implementation, the cannula comprises a glass fiber.

[0008] According to another configuration of the above implementation, the polymeric support material is tapered, i.e., the polymeric support material tapers from the proximal end to the distal end, with the thickness of the polymeric support material being greater at the proximal end.

[0009] According to a further configuration of the above implementation, the polymer support material comprises polytetrafluoroethylene (PTFE), polyamide, fluoropolymer, polyolefin, PVC (polyvinyl chloride), polyimide, PEEK (polyetheretherketone), or combinations thereof.

[0010] According to a further aspect of the above implementation, the polymeric support material is polymeric support tubing.

[0011] According to a further aspect of the above implementation, the polymeric support material completely surrounds the cannula.

[0012] Moreover, in accordance with further aspects of the above implementation, the hub is a Luer interference fit hub. The Luer interference fit hub may include thread formations configured to be attached to a syringe.

[0013] According to a further aspect of the above implementation, the cannula assembly further includes a winged connection adapted to clamp the cannula assembly and the syringe.

[0014] In another aspect of the above implementation, the polymeric support material and the hub are made of a transparent or translucent material.

[0015] According to a further aspect of the above implementation, the adhesive is a UV-sensitive adhesive.

[0016] In another aspect of the above implementation, the cannula assembly further includes upper tubing, the upper tubing disposed adjacent the hub opposite from the polymeric support material.

[0017] Moreover, according to a further aspect of the above implementation, the polymeric support material is a plurality of polymeric support tube segments.

[0018] According to a further aspect of the present disclosure, a cannula assembly and syringe combination includes a cannula and a syringe. The cannula has a proximal end and a distal end. The polymeric support material substantially surrounds a portion of the cannula at or near the proximal end. The polymeric support material is disposed between the cannula and a hub. The polymeric support material and the hub are attached by adhesive. The syringe includes a needle. The hub attaches the cannula and the syringe.

[0019] According to one configuration of the above implementations, the polymeric support material is tapered, i.e., the polymeric support material tapers from the proximal end to the distal end, with the thickness of the polymeric support material being greater at the proximal end.

[0020] According to a further aspect of the present disclosure, the polymeric support material completely surrounds the cannula.

[0021] According to one configuration of the above implementation, the hub is a Luer interference fit hub.

[0022] According to one method of the present disclosure, a cannula assembly is formed. A cannula is provided having proximal and distal ends, a polymeric support material, and a hub. The polymeric support material and the hub comprise a transparent or translucent material. The polymeric support material is disposed to substantially surround a portion of the cannula at or near the proximal end. The polymeric support material is disposed between the cannula and the hub. An adhesive is placed on at least one of the cannula, the polymeric support material, and the hub. The adhesive is exposed to ultraviolet light to securely attach the cannula, the polymeric support material, and the hub.

[0023] According to a further method of the present disclosure, the polymeric support material is tapered, i.e., the polymeric support material tapers from the proximal end to the distal end, with the thickness of the polymeric support material being greater at the proximal end.

[0024] According to one configuration of the above method, the polymeric support material completely surrounds the cannula.

[0025] According to one configuration of the above method, the hub is a Luer interference fit hub.

[0026] According to a further method of the present disclosure, a cannula assembly is formed. A cannula is provided having proximal and distal ends, a polymeric support material, and a hub. The polymeric support material is disposed to substantially surround a portion of the cannula at or near the proximal end. The polymeric support material is disposed between the cannula and the hub. The hub and polymeric support material are shrink wrapped about the cannula to securely attach the cannula, the polymeric support material, and the hub.

[0027] According to yet another method, a viral vector is delivered to the central nervous system of a subject. A cannula assembly and syringe combination is provided, including the cannula assembly and the syringe. The cannula assembly includes a cannula, a polymeric support material, and a hub. The cannula has a proximal end and a distal end. The polymeric support material substantially surrounds a portion of the cannula at or near the proximal end. The syringe includes a needle. The hub attaches the cannula assembly and the syringe. The polymeric support material is disposed between the cannula and the hub. The cannula, the polymeric support material, and the hub are attached by adhesive. A viral vector is provided. The viral vector is delivered to the central nervous system via the cannula assembly and syringe combination.

[0028] In further aspects of the above methods, the viral vector is a recombinant viral vector.

[0029] In further aspects of the above methods, the viral vector has a dosage of about 0.5E9 to about 1.5E9 vg / μl. The viral vector can have a dosage of about 0.7E9 to about 1.3E9 vg / mil, or about 0.7E9 to about 1.1E9 vg / mil.

[0030] In another aspect of the above method, the central nervous system is brain tissue or spinal cord.

[0031] In a further aspect of the above method, the subject has a neurological disorder.The neurological disorder can be meningitis, encephalitis, multiple sclerosis (MS), stroke, brain tumor, epilepsy, Alzheimer's disease, AIDS dementia, Parkinson's disease, or Huntington's disease.More specifically, the neurological disorder can be Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0032] The above summary is not intended to represent each embodiment or every aspect of the present disclosure. Rather, the foregoing summary provides only some examples of some of the novel aspects and features described herein. The above features and advantages, as well as other features and advantages of the present disclosure, will be readily apparent from the following detailed description of exemplary embodiments and modes for carrying out the invention, when considered in conjunction with the accompanying drawings and the appended claims. Additional aspects of the present disclosure will become apparent to those skilled in the art in light of the detailed description of various embodiments, which are provided with reference to the drawings, a brief description of which is provided below. [Brief description of the drawings]

[0033] The present disclosure and its advantages and drawings will be better understood from the following description of exemplary embodiments with reference to the accompanying drawings, which depict only exemplary embodiments and therefore should not be considered as limitations on the scope of the various embodiments or claims.

[0034] [Figure 1A] FIG. 1A is a cross-sectional side view of a cannula assembly according to one embodiment.

[0035] [Figure 1B] FIG. 1B is a front view of the cannula assembly of FIG. 1A.

[0036] [Figure 1C]FIG. 1C is an enlarged side view of area 1C taken from FIG. 1A with adhesive added.

[0037] [Figure 2A] FIG. 2A is a cross-sectional side view of a cannula assembly according to another embodiment.

[0038] [Figure 2B] FIG. 2B is an enlarged side view of area 2B taken from FIG. 2A.

[0039] [Diagram 3] FIG. 3 is a cross-sectional side view of a cannula assembly according to a further embodiment.

[0040] [Figure 4] FIG. 4 is a perspective view of a syringe according to one embodiment.

[0041] [Figure 5A] 5A is a cross-sectional side view of the cannula assembly of FIG. 1 in combination with the syringe of FIG. 4. FIG.

[0042] [Figure 5B] FIG. 5B is an enlarged side view of area 5B taken from FIG. 5A. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0043] While the invention is susceptible to various modifications and alternative forms, specific implementations have been shown by way of example in the drawings and will be described in more detail herein. It should be understood, however, that the invention is not intended to be limited to the particular forms disclosed. Rather, the invention is intended to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention as defined by the appended claims.

[0044] Various embodiments are described with reference to the attached figures, in which like reference numerals are used throughout the figures to designate similar or equivalent elements. The figures are not drawn to scale and are provided merely to illustrate the present invention. Several aspects of the present invention are described below with reference to example applications for illustration. It should be understood that numerous specific details, relationships, and methods are set forth to provide a thorough understanding of the present invention. However, one skilled in the art will readily recognize that the present invention can be practiced without one or more of the specific details, or with other methods. In other instances, well-known structures or operations have not been shown in detail to avoid obscuring the present invention. Various embodiments are not limited by the illustrated order of acts or events, as some acts may occur in different orders and / or in parallel with other acts or events. Furthermore, not all illustrated acts or events are required to implement a methodology in accordance with the present invention.

[0045] 1A and 1B, a cannula assembly 10 according to one embodiment is shown in cross-sectional side and front views, respectively. Cross-hatched lines have been removed to improve clarity in FIG. 1A. The cannula assembly of the present invention, in one embodiment, is configured to engage with a syringe (e.g., syringe 60 shown in FIG. 4). The cannula assembly assists in delivering liquid products to areas of the body including brain tissue. The cannula assembly allows for a more practical and leak-proof connection to a delivery syringe for brain infusion. The cannula assembly also assists in removing liquids or collecting samples from areas of the body including from brain tissue. It is envisioned that the cannula assembly may be used in other aspects in other implementations.

[0046] 1A and 1B, the cannula assembly 10 includes a cannula 20, a polymeric support material 30, and a hub 40. FIG. 1C is an enlarged side view of area 1C taken from FIG. 1A with adhesive added. The cannula 20 is a tube in one embodiment and is configured to be inserted into the body. The cannula 20 in one embodiment is a fiberglass cannula. It is envisioned that the cannula may be made from other materials, such as metals. Non-limiting examples of metals that may be used in forming the cannula include stainless steel and titanium. It is also envisioned that a polymeric material with a desired stiffness may be used in forming the cannula.

[0047] The overall length of the cannula 20 is truncated in FIG. 1A for clarity and therefore not shown to scale. The cannula has a proximal end 22 and a distal end 24. The distal end 24 is configured to be inserted into the body. The cannula 20 can vary in length, but typically has a length L1 of about 0.3 to about 1.5 meters. In one desired embodiment, the cannula 20 has a length L1 of about 0.75 to about 1.25 meters. In another desired embodiment, the cannula 20 has a length L1 of about 0.9 to about 1.1 meters.

[0048] Cannula 20 can vary in diameter, but typically has a diameter D1 (see FIG. 1B) of about 0.25 to about 0.5 mm. In one desired embodiment, cannula 20 has a diameter D1 of about 0.3 to about 0.45 mm. In another desired embodiment, cannula 20 has a diameter D1 of about 0.3 to about 0.4 mm.

[0049] As shown in FIGS. 1A-1C, the polymeric support material 30 is disposed between the cannula 20 and the hub 40. Specifically, the polymeric support material as best shown in FIG. 1C has an inner surface 32 disposed adjacent the cannula 20 and an outer surface 34 disposed adjacent the hub 40. With reference to FIG. 1A, the polymeric support material 30 is disposed at or near the proximal end 22 of the cannula 20. As shown in FIGS. 1A-1C, the polymeric support material 30 substantially surrounds a portion of the cannula 20 at or near the proximal end 22. For example, the polymeric support material surrounds 70% or 85% of the portion of the cannula at or near the proximal end. In another embodiment, the polymeric support material surrounds 90% or 95% of the portion of the cannula at or near the proximal end. It is desirable for the polymeric support material 30 of FIGS. 1A-1C to completely surround a portion of the cannula 20 at or near the proximal end 22, as best seen in FIG. 1B.

[0050] The polymeric support material 30 helps to prevent or inhibit stress risers, which are sudden changes in flexibility that can likely increase fracturing of the cannula when the cannula is made from fiberglass. The polymeric support material 30 helps to change the stress points with the use of distance, which helps to prevent or inhibit fracturing of the cannula 20.

[0051] The polymeric support material 30 can vary in length, but typically has a length L2 (see FIG. 1A) of about 5 to about 40 cm. In one desired embodiment, the polymeric support material 30 has a length L2 of about 10 to about 30 cm. In another desired embodiment, the polymeric support material 30 has a length L2 of about 10 to about 20 cm or about 15 to about 20 cm.

[0052] The polymeric support material 30, in one embodiment, tapers from the proximal end 22 to the distal end 24 of the cannula 20. The thickness T1 of the polymeric support material 30 is greater at or near the proximal end 22 of the cannula 20. The thickness T1 of the polymeric support material 30 is from about 1 mm to about 5 mm, more specifically, from about 1.5 mm to about 4 mm, or from about 2 mm to about 4 mm.

[0053] In one embodiment, the polymeric support material 30 comprises polytetrafluoroethylene (PTFE). The polymeric support material may be made from other polymeric materials or combinations of polymeric materials. Some other non-limiting materials that may be used in forming the polymeric support material include polyamide, fluoropolymer, polyolefin, PVC (polyvinyl chloride), polyimide, PEEK (polyetheretherketone), or combinations thereof. It is also desirable that the polymeric support material is generally clear or translucent to allow transmission of ultraviolet (UV) light if a UV adhesive is used. It is also desirable to have the material that forms the polymeric support material compatible with adeno-associated virus (AAV) (i.e., the AAV virus does not adhere to the material).

[0054] 1A-1C, is disposed adjacent the exterior surface 34 of the polymeric support material 30. The hub 40 is configured to aid in attaching and securing the cannula 20 and syringe (e.g., syringe 60 of FIG. 4). The hub 40 desirably also provides a seal with the syringe that helps prevent or inhibit leakage of any liquid material contained within the syringe or within the proximal end 22 of the cannula 20, as will be discussed below. The hub 40 is also desirably configured to reduce or effectively eliminate much of the dead space where any liquid may not be accounted for or where air bubbles may accumulate.

[0055] 1A depicts a first generally horizontal section 42, a slightly upwardly tapered section 44, and a second generally horizontal section 46. The second generally horizontal section 46 includes an outer periphery 48. The outer periphery 48 reinforces the hub 40.

[0056] In one embodiment, hub 40 is a Luer interference fit hub. Some advantages of using a Luer interference fit hub include ease and safety of installation. Other hubs are also envisioned for use in the cannula assembly.

[0057] The hub 40 can be made from materials including, but not limited to, polymeric materials. Some non-limiting examples of polymeric materials include, but are not limited to, polyolefins (e.g., polypropylene). It is also desirable for the hub to be generally clear or translucent to allow transmission of UV light if a UV adhesive is used. It is also desirable to have the material forming the hub be compatible with adeno-associated virus (AAV).

[0058] The cannula 20, the polymeric support material 30, and the hub 40 are attached by adhesive in one embodiment. The adhesive may be applied and positioned in different areas to securely attach the cannula 20, the polymeric support material 30, and the hub 40. The adhesive is applied on at least one of the cannula, the polymeric support material, and the hub. It is envisioned that the adhesive may be applied on two or more of the cannula, the polymeric support material, and the hub. Some representative areas are shown in FIG. 1C with adhesive 70a-70d. The adhesive areas 70a, 70d are positioned between the hub 40 and the polymeric support material 30. The adhesive areas 70b, 70c are positioned between the polymeric support material 30 and the cannula 20. The adhesive permanently and securely attaches the cannula 20, the polymeric support material 30, and the hub 40.

[0059] In one embodiment, the adhesive is a UV sensitive adhesive. In this embodiment, the adhesive is a liquid adhesive that is typically cured using UV light. The surface tension of the material that the liquid adhesive comes into contact with helps to maintain the position of the adhesive before it cures. This can be done in a single step in one method. In another method, the cure of the adhesive can be made in a multi-step process.

[0060] UV adhesives are typically adhesives that work with epoxy resins or acrylic materials. One type of UV adhesive is a radical-initiated UV adhesive based on an acrylate mixture (e.g., urethane, cyanoacrylate, or silicone). Thus, UV adhesives include, but are not limited to, acrylate-urethane adhesive compositions, cyanoacrylate adhesive compositions, and acrylate-silicone adhesive compositions. Acrylate-based UV adhesives are typically solvent-free and contain one component. Another type of UV adhesive is a cationic-initiated UV adhesive based on epoxy resins. Notable UV adhesives are from companies such as Bondic, RapidFix, and Dymax.

[0061] To cure, UV-sensitive adhesives require a light source. This can come from pure sunlight, but also UV LED lights and gas discharge lamps. However, LED light sources must be matched to the respective adhesive and are therefore available in different wavelengths. UV-sensitive adhesives usually cure very quickly. For example, the curing of a UV-sensitive adhesive can occur in about 1 to about 10 seconds, more specifically, about 1 to about 5 seconds, and about 1 to about 3 seconds. Typically, the more intense the light source, the faster the curing process. A UV-sensitive adhesive can be an adhesive that cures only when the user exposes it to a precisely defined wavelength of UV light.

[0062] The cannula 20, the polymeric support material 30, and the hub 40 are securely attached such that an individual cannot easily separate the components from one another. Thus, for example, the cannula 20, the polymeric support material 30, and the hub 40 are not attached by an interference fit. Thus, the cannula 20, the polymeric support material 30, and the hub 40 are formed without an interference fit.

[0063] In one embodiment, the cannula assembly is a non-removable closed sterile system. In one method, products to be delivered using the cannula assembly do not come into contact with the adhesives used to securely attach the cannula 20, the polymeric support material 30, and the hub 40. Similarly, in another method, fluids or other products to be removed, or products to be delivered using the cannula assembly do not come into contact with the adhesives used to securely attach the cannula 20, the polymeric support material 30, and the hub 40.

[0064] Cannula assembly 10 may further include a winged connection 50 adapted to tighten the connection between a syringe (e.g., syringe 60 of FIG. 4) and the remainder of cannula assembly 10. In one process, winged connection 50 may be tightened by use of the thumb and index finger. To aid in gripping winged connection 50, it may include a wavy, concave area that generally matches the shape of the thumb or other finger.

[0065] 2A and 2B, a cannula assembly 110 according to another embodiment is shown in cross-sectional side view. Cross-hatching has been removed in FIGS. 2A and 2B for improved clarity. Cannula assembly 110 includes cannula 20, polymeric support material 30, hub 140, upper tubing 180, and winged connector 50.

[0066] Hub 140 is the same as hub 40 described above, except for outer rim 148. Outer rim 148 reinforces hub 140 in a manner similar to outer rim 48 for hub 40. Outer rim 148 also includes external thread formation 148a. External thread formation 148a can be a single thread or multiple threads. External thread formation 148a is configured to securely attach with a component of a syringe having an internal thread formation. For example, external thread formation 148a can be attached to a Hamilton connector ring of the syringe. Thus, external thread formation 148a aids in securely attaching cannula assembly 110 with a syringe (e.g., syringe 60 of FIG. 4).

[0067] The upper layer tubing 180 is disposed on the exterior surface 142 of the hub 140 as shown in FIGS. 2A, 2B. The upper layer tubing 180 in one embodiment is made of a flexible material. Non-limiting examples of materials that may be used in forming the upper layer tubing 180 include, but are not limited to, polytetrafluoroethylene (PTFE). Some other non-limiting materials that may be used in forming the upper layer tubing include polyamide, fluoropolymer, polyolefin, PVC (polyvinyl chloride), polyimide, PEEK (polyetheretherketone), or combinations thereof. It is also desirable that the polymeric support material is generally clear or translucent to allow transmission of UV light if a UV adhesive is used. The upper layer tubing 180 is disposed generally above the proximal end 22 of the cannula 20. The upper layer tubing 180 helps absorb and dissipate shear loads to prevent or inhibit breakage of the cannula 20.

[0068] The upper layer tubing 180 can vary in length, but typically has a length L3 (see FIG. 2A) of about 5 to about 25 cm. In one desired embodiment, the upper layer tubing 180 has a length L3 of about 10 to about 20 cm. In another desired embodiment, the upper layer support 180 has a length L3 of about 10 to about 15 cm.

[0069] 3, a cannula assembly 210 according to a further embodiment is shown in cross-sectional side view. Cross-hatching has been removed in FIG. 3 for improved clarity. Cannula assembly 210 includes cannula 20, polymeric support material 230, hub 40, and winged connector 50. Polymeric support material 230 helps prevent or inhibit stress risers.

[0070] The polymer support material 230 includes a plurality of polymer support tube segments 232, 234, 236, 238, 240 stacked on top of one another in series. The lengths L4-L8 (see FIG. 3) of each of the plurality of polymer support tube segments 232, 234, 236, 238, 240 are different. The length L4 of the polymer support tube segment 232 is the longest, while the length L8 is the shortest. Each of the lengths of the polymer support tube segments 232, 234, 236, 238, 240 is progressively shorter. The length L4 of the polymer support tube segment 232 is similar to the length L1 of the polymer support material 30. It is contemplated that the number of polymer support tube segments may be greater or less than that depicted in FIG. 3.

[0071] As discussed above, the cannula assembly is adapted to work in conjunction with a syringe to deliver or receive fluids. One type of syringe that may be used with the cannula assembly is shown in FIG. 4 with a syringe 60. The syringe 60 includes a needle 62, a barrel assembly 64, a plunger assembly 66, and a male end 68. The needle 62 is typically covered by a nut (not shown) to protect against unintentional contact with the needle 62. The syringe 60 may be referred to as a Hamilton syringe. With reference to FIG. 4, the syringe 60 is a Hamilton syringe. It is envisioned that other types of syringes, such as those manufactured by Setonic, Trident, Becton Dickenson, and Alibaba, may also be used with the cannula assembly of the present invention.

[0072] 5A, 5B, a cannula assembly and syringe combination 300 is shown in cross-section. Cross-hatching has been removed in FIGS. 5A, 5B for improved clarity. The combination 300 includes the cannula assembly 10 described above and a syringe 60. The needle 62 of the syringe is not shown for improved clarity. To reduce dead space, the cannula 20 extends sufficiently into the hub 40 to receive the syringe 60. The cannula 20 should extend a sufficient distance beyond the polymer support material 30 such that process tolerances will not allow any potential adhesive to be applied near the opening 26 of the proximal end 22 of the cannula 20. The adhesive delivery and processing steps with UV light need to be precise to avoid plugging the cannula or creating leaks.

[0073] Because the proximal end 22 of the cannula 20 is disposed inside the lumen of the male end 68 of the syringe 60, minimal dead space exists. As shown in Figures 5A and 5B, dead spaces 90, 92 are shown between the male end 68, the hub 40, and the polymeric support material 30. The hub 40 desirably also provides a seal with the male end 68 of the syringe 60 to help prevent or inhibit leakage of any liquid material contained within the syringe or within the proximal end 22 of the cannula 20.

[0074] It is envisioned that combinations may be formed using cannula assemblies 110, 210 with syringe 60 or other syringes.

[0075] According to yet another method, a viral vector is delivered to the central nervous system of a subject. A cannula assembly and syringe combination is provided, including the cannula assembly and the syringe. The cannula assembly includes a cannula, a polymeric support material, and a hub. The cannula includes a proximal end and a distal end. The polymeric support material substantially surrounds a portion of the cannula at or near the proximal end. The syringe includes a needle. The hub attaches the cannula assembly and the syringe. The polymeric support material is disposed between the cannula and the hub. The cannula, the polymeric support material, and the hub are attached by adhesive. A viral vector is provided. The viral vector is delivered to the central nervous system via the cannula assembly and syringe combination.

[0076] Non-limiting examples of pharmaceuticals and therapeutic devices that can be delivered through a cannula assembly and syringe combination as described herein (e.g., cannula assembly and syringe combination 300) include, but are not limited to, drugs, nanoparticles, biological agents (e.g., cells, viruses, etc.).

[0077] In one embodiment, the vector can be, but is not limited to, a non-viral vector or a viral vector. In one embodiment of any aspect, the vector is a DNA or RNA virus. Non-limiting examples of viral vectors include AAV vectors, adenovirus vectors, lentivirus vectors, retrovirus vectors, herpesvirus vectors, alphavirus vectors, poxvirus vectors, baculovirus vectors, and chimeric virus vectors. Non-limiting examples of AAV include AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, and any chimera thereof. In some embodiments, the AAV is AAV rhesus serotype (AAV rh). Non-limiting examples of AAV rh serotypes are AAV rh8, AAV rh10, AAV rh20, AAV rh74, AAV rh39, AAV rh43, AAV rh38, AAV rh40, AAV rh2, AAV rh25, AAV rh57, AAV rh50, AAV rh49, AAV rh58, AAV rh61, AAV rh52, AAV rh53, AAV rh51, AAV rh64, AAV rh8, AAV rh1, AAV rh62, AAV rh48, AAV rh54, AAV rh55, AAV rh35, AAV rh37, AAV rh36, AAV rh13, AAV rh32, AAV rh33, and AAV rh34.

[0078] Any viral vector known in the art can be used with the cannula assembly and syringe combination. Examples of such viral vectors include, but are not limited to, Adenoviridae, Birnaviridae, Bunyaviridae, Caliciviridae, Capillovirus, Carlavirus, Carmovirus, Calimovirus, Closterovirus, Comelina yellow spot virus, Comovirus, Coronaviridae, PM2 phage, Corsicoviridae, Latent virus, Cryptovirus, Cucumoviridae ([PHgr]6 phage), Cystoviridae, Carnation ringspot, Dianthovirus, Broad bean wilt, Fabavirus, Filoviridae, Flaviviridae, Furovirus, Germinivirus, Giardiavirus, Hepadnaviridae, Herpesviridae, Hordeivirus, Iraluvirus, Inoviridae, Iridoviridae, Leviviridae, Liposthrixviridae, Luteovirus, Malafviridae ... Vectors derived from the genera Myoviridae, Maize chlorotic dwarf virus, Microviridae, Myoviridae, Necrovirus, Nepoviridae, Nodaviidae, Orthomyxoviridae, Papovaviridae, Paramyxoviridae, Parsnip yellow spot virus, Partitiviridae, Parvoviridae, Pea gill mosaic virus, Phycodnaviridae, Picornaviridae, Plasmaviridae, Prodoviridae, Polydnaviridae, Potexviridae, Potyvirus, Poxviridae, Reoviridae, Retroviridae, Rhabdoviridae, Rigidiovirus, Siphoviridae, Sobemovirus, SSV1 phage, Tectiviridae, Tenuivirus, Tetraviridae, Tobamovirus, Tobravirus, Togaviridae, Tombusvirus, Torovirus, Tyviridae, Tymovirus, and plant virus satellites.

[0079] An effective amount of a viral vector (e.g., a recombinant viral vector (rAAV)) is an amount sufficient to target an animal to infect or target a desired tissue. In some embodiments, an effective amount of a viral vector (e.g., a recombinant viral vector (rAAV)) is an amount sufficient to produce a stable somatic transgenic animal model. The effective amount depends mainly on factors such as the species, age, weight, health of the subject, and the tissue to be targeted, and therefore may vary among animals and tissues.

[0080] In some embodiments, a dose of a viral vector (e.g., a recombinant viral vector (rAAV)) is administered to a subject no more than once per calendar day (e.g., 24 hours). In some embodiments, a dose of a viral vector is administered to a subject no more than once per 2, 3, 4, 5, 6, or 7 calendar days. In some embodiments, a dose of a viral vector is administered to a subject no more than once per calendar week (e.g., 7 calendar days). In some embodiments, a dose of a viral vector is administered to a subject no more than every other week (e.g., once every 2 calendar weeks). In some embodiments, a dose of a viral vector is administered to a subject no more than once per calendar month (e.g., once every 30 calendar days). In some embodiments, a dose of a viral vector is administered to a subject no more than once per 6 calendar months. In some embodiments, a dose of a viral vector is administered to a subject no more than once per calendar year (e.g., 365 days or 366 days in a leap year).

[0081] Effective amount, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the minimum effective dose and / or maximum tolerated dose. The dosage can vary depending on the dosage form employed and the route of administration utilized. The therapeutically effective dose can be estimated initially from cell culture assays. The dose can also be formulated in animal models to achieve a dosage range between the minimum effective dose and the maximum tolerated dose. The effect of any particular dosage can be monitored by suitable bioassays, for example, assays related to neuronal degradation or functionality, among others. The dosage can be determined by a physician and adjusted as necessary to suit the observed effect of treatment.

[0082] In further aspects of the above method, the viral vector has a dosage of about 0.5E9 to about 1.5E9 vg / μl (about 0.5E12 to about 1.5E12 vg / mil). The viral vector can have a dosage of about 0.7E9 to about 1.3E9 vg / μl (about 0.7E12 to about 1.3E12 vg / mil). In another embodiment, the viral vector can have a dosage of about 0.7E9 to about 1.1E9 vg / μl (about 0.7E12 to about 1.1E12 vg / mil). The viral vector can be delivered to the subject. The desired dosage should be within the range between the minimum effective dose at the lower end and below the mild toxicity level.

[0083] Dose calculations can be complex, but are mandated by tissue reactions to the product delivered directly to the parenchyma of the striatum or another part of the brain. If the concentration is too low (volume too small), there is no therapeutic effect. If it is more concentrated, the product may be effective. If the concentration is too high, it will initiate an inflammatory response, and if strong enough, it will kill cells at the injection site. After the dose concentration is determined, the volume of that dilution that will be injected into each of the four striatal lobes is calculated. In humans and non-human primates (NHPs), volumes are measured individually (two caudates and two putamen) using MRI. The volume of product to be injected into each lobe is calculated as a percentage of the measured organ volume. For example, this percentage can range from about 15% to about 50%. If there is insufficient perfusion into the target lobe, a higher percentage (e.g., 35-50%) can be used. The typical healthy volumes for the putamen and caudate in human subjects are approximately 3.57 and 2.73 cm, respectively, in NHPs. 3 、 and about 0.55 and about 0.41 cm 3 It is.

[0084] In mice, a standard volume for the striatum can be used. In mice, the striatum is about 20 to about 37 mm. 3 For example, the volume delivered to a mouse may be 2 μL or 4 μL. In another example, the volume delivered to an NHP will be in a volume range of about 20 μL to about 250 μL m. In a further example, the volume delivered to a human will be in a volume range of about 140 μL to about 1 mL.

[0085] A single dose is delivered using the cannula assembly of the present invention. For example, needle delivery into the brain parenchyma requires overcoming the static fluid pressure of the tissue. Thus, in one method, a very slow rate of infusion is first used with an increasing rate through two stages to complete the delivery. This is generally referred to as "convection-enhanced delivery". It is envisioned that other modified versions of delivery, including other three stage infusion rates, may also be used.

[0086] The viral vector is delivered to the subject using a combination of a cannula assembly and a syringe. For example, the recombinant viral vector, preferably suspended in a physiologically compatible carrier (i.e., in a composition), can be administered to a subject, i.e., a host animal such as a human, mouse, rat, cat, dog, sheep, rabbit, horse, cow, goat, pig, guinea pig, hamster, chicken, turkey, or non-human primate (e.g., macaque). In some embodiments, the host animal does not include a human.

[0087] Subjects to which the methods of the present disclosure are applicable include veterinary subjects (e.g., dogs, cats, horses, etc.), and laboratory animal subjects (e.g., mice, rats, rabbits, pigs, goats, sheep, primates, etc.), and human subjects. The methods are applicable to all primates, including apes. In some embodiments, the methods are applied to humans. In other embodiments, the methods are applied to non-human primates.

[0088] Any desired area of ​​a subject can be targeted according to the methods described herein. In some cases, the desired area can be tissue, including, but not limited to, tissue of endodermal origin, tissue of ectodermal origin, and tissue of mesodermal origin. Nervous tissue is typically targeted. In some cases, nervous tissue of the central nervous system (CNS), including, for example, brain tissue and spinal cord tissue, can be targeted. In some cases, nervous tissue of the peripheral nervous system can be targeted. It is envisioned that other tissues can also be targeted according to the methods described herein.

[0089] In some cases, the methods may be applied for effective delivery / localization of agents to regions of interest within the mammalian nervous system, including the central or peripheral nervous system. Essentially any region of interest in the nervous system may be targeted according to the methods as described herein, including, but not limited to, the brain, spinal cord, spinal ganglia, etc.

[0090] In some cases, the methods may be applied for the effective delivery / localization of drugs to regions of interest in the mammalian brain. Essentially any region of interest in the brain may be targeted according to the methods as described herein.

[0091] The viral vectors described herein can be injected directly into any region of the brain, such as, for example, the occipital lobe, temporal lobe, parietal lobe, frontal lobe, cerebral cortex, cerebellum, hypothalamus, thalamus, pituitary gland, pineal gland, amygdala, hippocampus, and midbrain.

[0092] In some cases, one or more lobes or specific areas within a lobe may be targeted, including, but not limited to, the frontal lobe (either the entire frontal lobe or portions thereof including, but not limited to, the superior frontal, rostral midfrontal, caudal midfrontal, opercular, triangular, and orbital, lateral orbitofrontal, medial orbitofrontal, precentral, paracentral, frontal pole, combinations thereof, etc.), the parietal lobe (either the entire parietal lobe or portions thereof including, but not limited to, the superior parietal, inferior parietal, hyperlimbic, postcentral, precuneus, combinations thereof, etc.), the temporal lobe (either the entire temporal lobe or portions thereof including, but not limited to, the superior temporal, middle temporal, inferior temporal, bank of the superior temporal sulcus, fusiform, lateral temporal, entorhinal, temporal pole, parahippocampal, combinations thereof, etc.), and the occipital lobe (either the entire occipital lobe or portions thereof including, but not limited to, the lateral occipital, lingual, cuneus, pericalcarine, combinations thereof, etc.).

[0093] In some cases, the receptors may include, but are not limited to, hindbrain structures (e.g., myelencephalic structures (e.g., medulla oblongata, medullary pyramidal structures, olive bodies, inferior olivary nuclei, respiratory center, cuneate nucleus, gracilis nucleus, interneurons, medullary cranial nerve nuclei, inferior salivary nuclei, nucleus ambiguus, dorsal nucleus of the vagus, hypoglossal nucleus, solitary nucleus, etc.), hindbrain structures (e.g., pons, pontine cranial nerve nuclei, principal or pontine nuclei of the trigeminal sensory nucleus (V), motor nucleus of the trigeminal nerve (V), abducens nucleus (VI), facial nucleus (VII), vestibulocochlear nuclei (vestibular and cochlear nuclei) (VIII), superior salivary nucleus, pontine tegmentum, respiratory center, respiratory control center, tonic inspiration center, pontine micturition center (Burlington nucleus), locus coeruleus, etc.). , pedunculopontine nucleus, dorsolateral tegmental nucleus, reticular nucleus of the tegmental pons, superior olivary complex, paramedian pontine reticular formation, cerebellar peduncle, superior cerebellar peduncle, middle cerebellar peduncle, inferior cerebellar peduncle, fourth ventricle, cerebellum, vermis, cerebellar hemispheres, anterior lobe, posterior lobe, flocculonodular lobe, cerebellar nuclei, fastigial nucleus, intercalary nucleus, globular nucleus, embolic nucleus, dentate nucleus, etc.), midbrain structures (e.g., tectum, tectum tetrageminae, inferior colliculus, superior colliculus, pretectal area, tegmentum, periaqueductal gray matter, paracerebellar peduncular area, medial paracerebellar nucleus, lateral paracerebellar nucleus, subpeduncular nucleus (Kölliker-Fuse nucleus), rostral interstitial nucleus of the medial longitudinal fasciculus, midbrain reticular formation, dorsal raphe nucleus, red nucleus, ventral tegmental area, substantia nigra, pars compacta, pars reticularis, interpeduncular nucleus, cerebral stalk, cerebral peduncle, midbrain Cranial nerve nuclei, oculomotor nucleus (III), trochlear nucleus (IV), midbrain tract (midbrain aqueduct, aqueduct of Sylvius), etc.), anterior brain structures (e.g. diencephalon, epithalamic structures (e.g. pineal gland, habenula, stria medulla, thalamic tract, etc.), third ventricle, thalamic structures (e.g. anterior nuclei, anterior ventral nuclei (i.e. ventral anterior nuclei), anterior dorsal nuclei, anterior medial nuclei, medial nuclei, medial dorsal nuclei, midline nuclei, paranuclei, connective nuclei, rhomboid nuclei, intralaminar nuclei, centromedial nuclei, parafascicular nuclei, paracentral nuclei, lateral central nuclei, medial central nuclei, lateral nuclei, dorsal lateral nuclei, posterolateral nuclei, pulvinar, ventral nuclei, ventral anterior nuclei, ventral lateral nuclei, ventral posterior nuclei, ventral posterolateral nuclei, optic tract, posterior floor, medial geniculate body, lateral geniculate body, thalamic reticular nucleus, etc.), hypothalamic structures (e.g. anterior, medial region, part of preoptic area, medial preoptic nucleus, supraoptic nucleus, paraventricular nucleus, supraoptic nucleus (mainly), anterior hypothalamic nucleus, lateral region, part of preoptic area, lateral preoptic nucleus, anterior lateral nucleus, part of supraoptic nucleus, other nuclei of preoptic area, median preoptic nucleus, periventricular preoptic nucleus, eminence, medial region, dorsomedial nucleus of the hypothalamus, ventromedial nucleus, arcuate nucleus, lateral region, eminence of lateral nucleus, nucleus eminence lateralis, posterior, medial region, mammillary nucleus (part of mammillary body), posterior nucleus, lateral region, posterior lateral nucleus, optic chiasm, subfornical organ, periventricular nucleus, pituitary stalk, eminence gray, eminence nucleus,tuberomammillary nuclei, tuberal region, mammillary bodies, mammillary nuclei, etc.), ventral thalamic structures (e.g., thalamic nuclei, zona incerta, etc.), pituitary structures (e.g., neurohypophysis, pars intermedium (intermediate lobe), anterior pituitary gland, etc.), telencephalic structures, white matter structures (e.g., corona radiata, internal capsule, external capsule, outermost capsule, arcuate fasciculus, uncinate fasciculus, perforant pathway, etc.), subcortical structures (e.g., hippocampus (medial temporal lobe), dentate gyrus, cornu of Ammon (CA area), cornu of Ammon area region 1, cornu of Ammon area region 2, cornu of Ammon area region 3, cornu of Ammon area region 4, amygdala (limbic system) (limbic lobe), central nucleus (autonomic nervous system), medial nucleus (accessory olfactory system), cortex and basal ganglia (main olfactory system), lateral [necessary gyrus] avoidance] and the basolateral nucleus (frontotemporal system), claustrum, basal ganglia, striatum, dorsal striatum (i.e., neostriatum), putamen, caudate nucleus, ventral striatum, nucleus accumbens, olfactory tubercle, globus pallidus (together with the putamen to form the lenticular nucleus), subthalamic nucleus, basal anterior brain, substantia anterior foramen, substantia innominata, basal ganglia, Broca diagonal band, medial septal nucleus, etc.), olfactory brain structures (e.g., olfactory bulb, piriform cortex, anterior olfactory nucleus, olfactory tract, anterior commissure, uncus, etc.), brain cortical structures (e.g., frontal lobe, cortex, primary motor cortex (precentral gyrus, Ml), supplementary motor area, premotor cortex, prefrontal cortex, superior frontal gyrus, middle frontal gyrus, inferior frontal gyrus, Brodmann's areas (areas 4, 6, 8, 9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 0, 11, 12, 24, 25, 32, 33, 44, 45, 46, 47), parietal lobe, cortex, primary somatosensory cortex (S1), secondary somatosensory cortex (S2), posterior parietal lobe, gyrus, postcentral gyrus (primary somatosensory area), others, precuneus, Brodmann area (1, 2, 3 (primary somatosensory area), 5, 7, 23, 26, 29, 31, 39, 40), occipital lobe, cortex, primary visual cortex (V1), V2, V3, V4, V5 / MT, gyrus, lateral occipital gyrus, cuneus, Brodmann area (17 (V1, primary visual area), 18, 19), temporal lobe, cortex, primary auditory cortex one or more brain structures or specific areas within brain structures, including the auditory cortex (Al), secondary auditory cortex (A2), inferior temporal cortex, posterior inferior temporal cortex, superior temporal gyrus, middle temporal gyrus, inferior temporal gyrus, entorhinal cortex, perirhinal cortex, parahippocampal gyrus, fusiform gyrus, Brodmann's areas (areas 9, 20, 21, 22, 27, 34, 35, 36, 37, 38, 41, 42), middle superior temporal cortex (MST), insular cortex, cingulate cortex, anterior cingulate, posterior cingulate, retrosplenial cortex, gray white layer, subgenual area 25, and Brodmann's areas (areas 23, 24, 26, 29, 30 (retrosplenial area), 31, 32, etc.)Can be targeted.

[0094] In some cases, one or more neural pathways or specific portions of neural pathways may be targeted, including, but not limited to, the neural pathways of those lobes and structures described above, such as the superior longitudinal fasciculus, the arcuate fasciculus, the cerebral stalk, the corpus callosum, the pyramidal or corticospinal tracts, the major dopamine pathways of the dopamine system, the mesocortical pathways, the mesolimbic pathways, the nigrostriatal pathways, the tuberofundus pathways, the serotonin pathways of the serotonin system, the raphe nuclei, the norepinephrine pathway, and the locus coeruleus.

[0095] Diseased nervous tissue that may be targeted includes, but is not limited to, nervous tissue diseases resulting from one or more of meningitis, encephalitis, multiple sclerosis (MS), stroke, brain tumors, epilepsy, Alzheimer's disease, AIDS-related dementia, Parkinson's disease, and Huntington's disease.

[0096] Delivery of the composition to a mammalian subject can be by any known means, for example, to a desired site, e.g., the central nervous system (CNS). It can be desirable to deliver the composition to the CNS of a subject. By "CNS" is meant all cells and tissues of the vertebrate brain and spinal cord. Thus, the term includes, but is not limited to, neurons, glial cells, astrocytes, cerebrospinal fluid (CSF), interstitial spaces, bone, cartilage, and the like. Any of the compositions described herein can be delivered directly to the CNS or brain, for example, by injection into the ventricular region with a needle, catheter, or related device using neurosurgical techniques known in the art, such as by stereotactic injection, and into the striatum (e.g., the caudate or putamen of the striatum), spinal cord and neuromuscular junction, or cerebellar lobule. In some embodiments, the composition as described in this disclosure is administered by intravenous injection. In some embodiments, the composition as described in this disclosure is administered by intraspinal injection. In some embodiments, the composition as described in this disclosure is administered by intraventricular injection. In some embodiments, the composition is administered by intracerebral injection. In some embodiments, the composition is administered by intrathecal injection. In some embodiments, the composition is administered by intrastriatal injection. In some embodiments, the composition is delivered by intracranial injection. In some embodiments, the composition is delivered by cisternal injection. In some embodiments, the composition is delivered by lateral ventricle injection.

[0097] The CNS includes certain regions of the CNS, namely, but not limited to, neural pathways, the somatosensory system, the visual system, the auditory system, the nervous, neuroendocrine system, the neurovascular system, the brain neurotransmitter system, and the dural meningeal system.

[0098] Exemplary regions of the CNS include, but are not limited to, the myelencephalon, medulla oblongata, medullary pyramid, olivores, inferior olivary nucleus, rostral ventrolateral medulla, caudal ventrolateral medulla, solitary nucleus (nucleus tractus solitarii), respiratory center-respiratory group, dorsal respiratory group, ventral respiratory group or tonic inspiration center, pre-Bötzinger complex, Bötzinger complex, posterior trapezoid nucleus, posterior facial nucleus, nucleus ambiguus posterior, nucleus ambiguus paramedian reticular nucleus, nucleus gigantocellularis reticularis, parafacial zone, nucleus cuneate, nucleus gracilis, perineural hypoglossal nucleus, nucleus interna, nucleus prepositus, hypoglossal nucleus, area postrema, medullary cranial nerve nuclei, inferior salivary nucleus, nucleus ambiguus, dorsal nucleus of the vagus nerve, hypoglossal nucleus, chemoreceptor trigger zone, hindbrain, pons, pontine nuclei, pontine cranial nerve nuclei , principal nucleus of the trigeminal sensory nucleus or pontine nucleus, motor nucleus of the trigeminal nerve, abducens nucleus (VI), facial nucleus (VII), vestibular-cochlear nucleus (vestibular and cochlear nuclei) (VIII), superior salivary nucleus, pontine tegmentum, pontine micturition center (Burlington's nucleus), locus coeruleus, pedunculopontine nucleus, dorsolateral tegmental nucleus, reticular nucleus of the tegmentopontine, intercalary nucleus, paracerebellar peduncle, medial paracerebellar nucleus, lateral paracerebellar nucleus, paracerebellar subpeduncular nucleus (Kölliker-Fuse nucleus), pontine respiratory group, superior olivary complex, medial superior olive, lateral superior olive, medial nucleus of the trapezoid body, paramedian pontine reticular formation, parvocellular reticular nucleus, caudal pontine reticular nucleus, cerebellar peduncle, superior cerebellar peduncle, middle cerebellar peduncle, inferior cerebellar peduncle, fourth ventricle, cerebellum, cerebellum Vermis, cerebellar hemispheres, anterior lobe, posterior lobe, flocculonodular lobe, cerebellar nuclei, fastigial nucleus, intercalary nucleus, globular nucleus, embolized nucleus, dentate nucleus, midbrain, tectum, tectum, inferior colliculus, superior colliculus, pretectal area, tegmentum, periaqueductal gray matter, rostral interstitial nucleus of the medial longitudinal fasciculus, midbrain reticular formation, dorsal raphe nucleus, red nucleus, ventral tegmental area, parapeduncular pigmented nucleus, substantia nigra paranucleus, rostral medial tegmental nucleus, caudal linear nucleus, rostral linear nucleus of the raphe, interfascicular nucleus, substantia nigra, pars compacta, pars reticularis, interpeduncular nucleus, cerebral stalk, cerebral peduncle, mesencephalic nerve nucleus, oculomotor nucleus (III), Edinger-Westphal nucleus, trochlear nucleus (IV), mesencephalic duct (aqueduct of the midbrain, aqueduct of Sylvius), anterior brain, interbrain Brain, epithalamus, pineal gland (pineal gland), habenula, stria medulla, thalamic cord, third ventricle, subcommissural organ, thalamus, anterior nuclei, anterior ventral nuclei (i.e. ventral anterior nuclei), anterior dorsal nuclei, anterior medial nuclei, medial nuclei, medial dorsal nuclei, midline nuclei, paranucleus, connective nucleus, rhomboid nuclei, intralaminar nuclei, centromedial nuclei, parafascicular nuclei, paracentral nuclei, lateral central nuclei, lateral nuclei, dorsal lateral nuclei, posterolateral nuclei, pulvinar, ventral nuclei, ventral anterior nuclei, ventral lateral nuclei, ventral posterior nuclei, ventral posterolateral nuclei, posterior thalamus, medial geniculate body, lateral geniculate body, thalamic reticular nucleus, hypothalamus (limbic system) (HPA axis), anterior, medial region, part of preoptic area, medial preoptic nucleus, INAH1,INAH2, INAH3, INAH4, median preoptic nucleus, suprachiasmatic nucleus, paraventricular nucleus, supraoptic nucleus (mainly), anterior hypothalamic nucleus, lateral region, part of preoptic area, lateral preoptic nucleus, anterior lateral nucleus, part of supraoptic nucleus, other nuclei of preoptic area, median preoptic nucleus, periventricular preoptic nucleus, eminence, medial region, dorsomedial hypothalamic nucleus, ventromedial nucleus, arcuate nucleus, lateral region, eminence of lateral nucleus, nucleus eminence lateralis, posterior, medial region, mammillary nucleus (part of mammillary body), posterior nucleus, lateral region, posterior lateral nucleus, superficial, median eminence, mammillary body, pituitary stalk (infundibulum), optic chiasm, subfornical organ, periventricular nucleus, gray eminence, eminence nucleus, eminence mammillary body nucleus, eminence region, mammillary nucleus, Ventral thalamus (HPA axis), Subthalamic nucleus, Zona incerta, Pituitary gland (HPA axis), Neurohypophysis, Pars intermedium (intermediate lobe), Anterior pituitary gland, Telencephalon (cerebrum), Cerebral hemispheres, White matter, Centrum semiovale, Corona radiata, Internal capsule, External capsule, Outermost capsule, Subcortex, Hippocampus (medial temporal lobe), Dentate gyrus, Horn of Ammon (CA area), Horn of Ammon area 1 (CA1), Horn of Ammon area 2 (CA2), Horn of Ammon area 3 (CA3), Horn of Ammon area 4 (CA4), Amygdala (limbic system) (limbic lobe), Central nucleus (autonomic nervous system), Medial nucleus (accessory olfactory system), Cortex and basal ganglia (main olfactory system), Lateral and basolateral nuclei (frontotemporal system), Extended amygdala, stria terminalis, bed nucleus of the stria terminalis, claustrum, basal ganglia, striatum, dorsal striatum (i.e., neostriatum), putamen, caudate nucleus, ventral striatum, nucleus accumbens, olfactory tubercle, globus pallidus (together with the putamen to form the lenticular nucleus), ventral globus pallidus, subthalamic nucleus, basal anterior brain, substantia foramen, substantia innominata, basal ganglia, diagonal band of Broca, septal nucleus, medial septal nucleus, lamina terminalis, organa vasculosa of the lamina terminalis, olfactory brain (old cortex), olfactory bulb, olfactory tract, anterior olfactory nucleus, piriform cortex, anterior commissure, uncinate, periamygdaloid cortex, cerebral cortex (neocortex), frontal lobe, cortex, primary motor cortex (precentral gyrus, M1), supplementary motor area, premotor cortex, prefrontal cortex, orbitofrontal cortex, dorsolateral prefrontal cortex, gyrus, superior frontal gyrus, middle prefrontal gyrus Temporal gyrus, inferior frontal gyrus, Brodmann area (areas 4, 6, 8, 9, 10, 11, 12, 24, 25, 32, 33, 44, 45, 46, 47), parietal lobe, cortex, primary somatosensory cortex (S1), secondary somatosensory cortex (S2), posterior parietal lobe, gyrus, postcentral gyrus (primary somatosensory area), Brodmann area (areas 1, 2, 3 (primary somatosensory area), 5, 7, 23, 26, 29, 31, 39, 40), occipital lobe, cortex, primary visual cortex (V1), V2, V3, V4, V5 / MT, gyrus, lateral occipital gyrus, Brodmann area (area 17 (V1, primary visual cortex), area 18,19), temporal lobe, cortex, primary auditory cortex (A1), secondary auditory cortex (A2), inferior temporal cortex, posterior inferior temporal cortex, superior temporal gyrus, middle temporal gyrus, inferior temporal gyrus, entorhinal cortex, perirhinal cortex, parahippocampal gyrus, fusiform gyrus, Brodmann's area (areas 20, 21, 22, 27, 34, 35, 36, 37, 38, 41, 42), insular cortex, cingulate cortex, anterior cingulate, posterior cingulate, retrosplenial cortex, gray layer, subgenual area 25, and Brodmann's area (areas 23, 24, 26, 29, 30 (retrosplenial area), 31, and 32).

[0099] Exemplary neural pathways include, but are not limited to, superior longitudinal fasciculus, arcuate fasciculus, uncinate fasciculus, perforant pathway, thalamocortical radiation, corpus callosum, anterior commissure, amygdala efferent pathway, interthalamic bridge, posterior commissure, habenular commissure, fornix, mammillary tegmentum, fasciculus, interthalamic pathway, cerebral stalk, medial anterior fasciculus, medial longitudinal fasciculus, myoclonus triangle, tractus solitarius, major dopaminergic pathway from dopaminergic cell groups, mesocortical pathway, mesolimbic pathway, nigrostriatal pathway, tuberofundus pathway, serotonergic pathway, raphe nucleus, norepinephrine pathway, locus coeruleus and other noradrenergic cell groups. , epinephrine pathways from adrenergic cell groups, glutamine and acetylcholine pathways from the middle pontine nucleus, motor system / descending fibers, extrapyramidal system, pyramidal tracts, corticospinal or cerebrospinal fibers, lateral corticospinal tract, anterior corticospinal tract, corticopontine fibers, frontopontine fibers, temporopontine fibers, corticobulbar tract, corticomesencephalic tract, tectospinal tract, interstitial nucleus-spinal tract, rubrospinal tract, rubro-olivary tract, olivo-cerebellar tract, olivo-spinal tract, vestibulospinal tract, lateral vestibulospinal tract, medial vestibulospinal tract, reticulospinal tract, lateral raphe-spinal tract, alpha system, and gamma system.

[0100] Exemplary somatosensory systems include, but are not limited to, the dorsal column-medial lemniscus pathway, gracilis, cuneate fasciculus, medial lemniscus, spinothalamic tract, lateral spinothalamic tract, anterior spinothalamic tract, spinomesencephalic tract, spinocerebellar tract, spinolivary tract, and spinoreticular tract.

[0101] Exemplary visual system components include, but are not limited to, the optic tract, the optic radiation, and the retinohypothalamic tract.

[0102] Exemplary auditory systems include, but are not limited to, the stria of the fourth ventricle, the trapezoid body, and the lateral lemniscus.

[0103] Exemplary nerves include, but are not limited to, brainstem, cranial nerves, terminalis (0), olfactory (I), visual (II), oculomotor (III), trochlear (IV), trigeminal (V), abducens (VI), facial (VII), vestibulocochlear (VIII), glossopharyngeal (IX), vagus (X), accessory (XI), and hypoglossal (XII).

[0104] Exemplary neuroendocrine systems include, but are not limited to, the hypothalamic-pituitary hormones, the HPA axis, the HPG axis, the HPT axis, and the GHRH-GH.

[0105] Exemplary neurovascular systems include, but are not limited to, the middle cerebral artery, the posterior cerebral artery, the anterior cerebral artery, the vertebral artery, the basilar artery, the circle of Willis (arterial system), the blood-brain barrier, the glymphatic system, the venous system, and the circumventricular organs.

[0106] Exemplary brain neurotransmitter systems include, but are not limited to, the noradrenergic system, the dopamine system, the serotonin system, the cholinergic system, GABA, neuropeptides, opioid peptides, endorphins, enkephalins, dynorphins, oxytocin, and substance P.

[0107] Exemplary dural meningeal systems include, but are not limited to, the cerebrospinal fluid barrier, meningeal coverings, dura mater, arachnoid mater, pia mater, epidural space, subdural space, subarachnoid space, superior cistern, cistern endplate, cistern chiasmus, cistern interpeduncular, pontine cistern, cisterna magna, spinal subarachnoid space, ventricular system, cerebrospinal fluid, third ventricle, fourth ventricle, lateral ventricle, angular bundle, anterior horn, body of the lateral ventricular, inferior horn, posterior horn, calcaneus, and subventricular zone.

[0108] "PNS" refers to the nerves and ganglia outside the brain and spinal cord. The main function of the PNS is to connect the CNS to the limbs and organs, essentially serving as a relay between the brain, spinal cord, and the rest of the body. Unlike the CNS, the PNS is not protected by the spinal column and skull, or by the blood-brain barrier, leaving it exposed to, for example, toxins and mechanical injury.

[0109] The peripheral nervous system (PNS) is divided into the somatic and autonomic nervous systems. In the somatic nervous system, the cranial nerves are part of the PNS, excluding the optic nerve (cranial nerve II) in addition to the retina. The second cranial nerve is not a true peripheral nerve, but is a cord of the diencephalon. The cranial ganglia arise within the CNS. However, the remaining ten cranial nerve axons extend beyond the brain and are therefore considered part of the PNS. The autonomic nervous system exerts involuntary control over smooth muscles and glands. The connections between the CNS and the organs allow the system to be in two different functional states: sympathetic and parasympathetic.

[0110] As used herein, a "neurological disease or disorder" can refer to any disease, disorder, or condition that affects or is associated with the nervous system, i.e., those that affect the central nervous system (brain and spinal cord), the peripheral nervous system (peripheral and cranial nerves), and the autonomic nervous system (portions located within both the central and peripheral nervous systems). Over 600 neurological diseases have been identified in humans. By way of non-limiting example, neurological diseases or disorders include Alzheimer's disease, Parkinson's disease, Huntington's disease, Canavan disease, Leigh's disease, spinal encephalomyelopathy, Krabbe disease, Batten disease, Refsum disease, Tourette's syndrome, primary lateral sclerosis, amyotrophic lateral sclerosis, progressive muscular atrophy, Pick's disease, muscular dystrophy, multiple sclerosis, myasthenia gravis, Binswanger's disease, trauma due to spinal cord or head injury, eye diseases and disorders, Tay-Sachs disease, Lesch-Nyhan disease, epilepsy, cerebral infarction, depression, bipolar affective disorder, persistent affective disorder, secondary mood disorder, schizophrenia, drug addiction, neurosis, psychosis, dementia, paranoia, attention deficit disorder, psychosexual disorder, sleep disorder, pain disorder, and / or eating or weight disorder. In some embodiments, the neurological disease or disorder is a central nervous system (CNS) disease or disorder, such as Huntington's disease, Parkinson's disease, or Alzheimer's disease.

[0111] In one aspect, the present invention provides a method of treating a neurological disorder involving the cortex, referred to herein as a "cortical neuropathy." The method involves delivery of a viral vector described herein or a combination thereof to the CNS or PNS. Preferred cortical neuropathy involves a large area of ​​the cortex, preferably involving two or more functional areas of the cortex, preferably involving two or more lobes of the cortex up to the entire cortex. Preferred cortical neuropathy includes, but is not limited to, traumatic brain injury, stroke, enzyme dysfunction, psychiatric disorders including post-traumatic stress syndrome, neurodegenerative diseases including Huntington's disease, Parkinson's disease, and Alzheimer's disease, epilepsy, and cognitive disorders including dementia, autism, and depression. Preferred enzyme dysfunction disorders include, but are not limited to, leukodystrophy, including Canavan's disease, and lysosomal storage diseases (LSDs) including Niemann-Pick disease, Gaucher disease, Batten disease, Fabry disease, and Pompe disease.

[0112] "Cortical neuropathy" as used herein refers to a neuropathy involving the cortex. A cortical neuropathy is a neuropathy that (i) involves some population of cells in the cortex that are directly anatomically connected to the thalamus and / or (ii) involves populations of cells that are directly anatomically connected to cortical cell populations in (i).

[0113] Preferred cortical neurological disorders are those involving large areas of the cortex, preferably two or more functional areas of the cortex, preferably two or more lobes of the cortex, and up to the entire cortex. Preferred cortical neurological disorders include, but are not limited to, traumatic brain injury, stroke, enzyme dysfunction, psychiatric disorders including post-traumatic stress syndrome, neurodegenerative diseases including Huntington's disease, Parkinson's disease, and Alzheimer's disease, epilepsy, and cognitive disorders including dementia, autism, and depression. Preferred enzyme dysfunction disorders include, but are not limited to, leukodystrophy, including Canavan's disease, and lysosomal storage diseases (LSDs), including Niemann-Pick disease, Gaucher disease, Batten disease, Fabry disease, and Pompe disease. This list of disorders is exemplary and non-limiting.

[0114] Based on cortical pathology and neuroanatomical connectivity, the neurological disorder suitable for treatment by the present method will be reasonably clear to those skilled in the art. "Cortex" as used herein refers to the brain cortex. In some embodiments, the neurological disease or disorder is a CNS disease or disorder, such as Huntington's disease, Parkinson's disease, or Alzheimer's disease. In some embodiments, the neurological disease or disorder is a PNS disease or disorder, such as peripheral neuropathy.

[0115] According to one method, a viral vector is delivered to the central nervous system of a subject. The method includes providing a cannula assembly and syringe combination including a cannula assembly and a syringe. The cannula assembly includes a cannula, a polymeric support material, and a hub. The cannula has a proximal end and a distal end. The polymeric support material substantially surrounds a portion of the cannula at or near the proximal end. The syringe includes a needle. The hub attaches the cannula assembly and the syringe. The polymeric support material is disposed between the cannula and the hub. The cannula, the polymeric support material, and the hub are attached by adhesive. A viral vector is provided and delivered to the central nervous system via the cannula assembly and syringe combination.

[0116] According to another method, a neurological disorder is treated in a subject in need thereof. The method includes providing a cannula assembly and syringe combination including a cannula assembly and a syringe. The cannula assembly includes a cannula, a polymeric support material, and a hub. The cannula has a proximal end and a distal end. The polymeric support material substantially surrounds a portion of the cannula at or near the proximal end. The syringe includes a needle. The hub attaches the cannula assembly and the syringe. The polymeric support material is disposed between the cannula and the hub. The cannula, the polymeric support material, and the hub are attached by adhesive. A viral vector is provided and delivered to the central nervous system via the cannula assembly and syringe combination. The neurological disorder is treated using the viral vector delivered to the central nervous system.

[0117] Neurological disorders in this method include, but are not limited to, meningitis, encephalitis, multiple sclerosis (MS), stroke, brain tumor, epilepsy, Alzheimer's disease, AIDS dementia, Parkinson's disease, or Huntington's disease.

[0118] In the present invention, the viral vector contains a therapeutic nucleic acid (e.g., DNA or RNA) in its genome. The rAAV can contain any nucleic acid that has a therapeutic benefit. In some embodiments, the therapeutic nucleic acid is non-coding. For example, the therapeutic nucleic acid is a non-coding RNA. Non-limiting examples of non-coding RNA are shRNA, siRNA, miRNA. In other embodiments, the therapeutic nucleic acid encodes a therapeutic transgene.

[0119] Non-limiting examples of therapeutic transgenes that may provide a therapeutic benefit for diseases or disorders of the CNS include CYP46A1 and HTT (for Huntington's disease), AADC and GDNF (for Parkinson's disease), GLB1 (for GM1), GDNF (for MSA), ASM (for Niemann-Pick disease), CYP46A1 (for Alzheimer's disease, ALS, MS, and epilepsy), and UBE3A (for Angelman disease). In some embodiments, the polypeptide-encoding transgene encodes an antibody or antigen-binding fragment thereof. Approaches for the treatment of CNS diseases or disorders can also be used, targeting metabolic pathways (e.g., CYP46A1 to clear protein lipid rafts or protein aggregates for Huntington's disease, Parkinson's disease, ALS, and Alzheimer's disease) (similar approaches could also target synuclein and / or tau), using miRNA, shRNA, and / or ribozyme mediated knockdown of undesired mRNA transcripts (e.g., mHTT or HTT for Huntington's disease, or ATS knockdown for Angelman's disease), transgene expression for gene replacement (e.g., to restore normal splicing in Huntington's disease by adding MBNL2 or SFRF6) (as well as more conventional gene replacement by expression of anti-synuclein antibodies, AADC, GDNF, or other transgenes). Diseases can include, among others, neurodegenerative diseases (Parkinson's, Huntington's, Alzheimer's, ALS, multiple sclerosis, epilepsy) and congenital mutations (AADC, Angelman, Niemann-Pick, MPS, and others). Transgene-mediated gene editing, including homologous recombination (which may be applied, for example, to Angelman), e.g., CRISPR, or ARCUS, or other gene editing techniques, are also envisioned.

[0120] Example 1

[0121] Delivery of recombinant viral vectors encoding AADC to the primate brain

[0122] A recombinant AAV vector encoding human AADC (AAV2-hAADC) is delivered to the putamen of rhesus monkeys as follows.

[0123] (Recombinant Vector Production)

[0124] Recombinant AAV2 is generated by a triple transfection protocol (Matsushita et al. (1998) Gene Ther. 5(7):938-45). Briefly, after propagation of cells from a HEK293 working cell bank through a series of disposable culture vessels in DMEM containing 10% fetal bovine serum and 2 mM glutamine, cells are cotransfected with three plasmids (pAAV-hAADC-2, pHLP19, and Pradeno5). The rAAV2-hAADC vector clone is the same as that previously described (Sanftner et al. (2004) Mol. Ther. 9(3):403-9). Plasmids pHLP19 and Pradeno5 are more fully described in U.S. Patent Nos. 5,139,941, 5,622,856, 6,001,650, and 6,004,797, the disclosures of which are incorporated herein by reference in their entireties.

[0125] After the appropriate transfection time, the medium containing the transfection reagent is replaced with serum-free medium and the cells are further incubated to allow vector production. The cells are harvested, concentrated by centrifugation, and lysed by a freeze / thaw method to release the AAV-hAADC-2 vector. After centrifugation to remove cell debris, the lysate is treated with Benzonase®, calcium chloride, and precipitated with polyethylene glycol. The vector is purified by two cycles of isopycnic gradient ultracentrifugation in cesium chloride. The AAV2-hAADC is concentrated and diafiltered against sterile buffered saline (PBS) containing 5% sorbitol. Poloxamer 188 TM(0.001%) is added and the material is sterile filtered (0.22 μm) and stored frozen at -70°C. Vector purity is assessed by SDS-PAGE. Purified rAAV2 vectors to be used in this study will only display VP1, VP2, and VP3 by silver staining of SDS-PAGE gels. Titers are determined by real-time Q-PCR analysis of the vector genome.

[0126] (Surgical Procedure)

[0127] Stereotaxic coordinates (based on the anatomy of the putamen) are first identified in the rhesus monkey. Two sites are targeted in each hemisphere with one site centered within the rostral putamen and the second centered within the caudal putamen. Adult rhesus monkeys (n=4) are immobilized, intubated, and prepared for surgery with a mixture of ketamine (Ketaset®, 10 mg / kg, intramuscular injection) and Valium® (0.5 mg / kg, intravenous injection). Isotonic fluid is delivered intravenously at 2 mL / kg / hr. Anesthesia is induced with isoflurane (Aerane®, Omeda PPD, Inc., Liberty, NJ) at 5% v / v, then maintained at 1%-3% v / v for the duration of the surgery. The animal's head is placed in an MRI-compatible stereotaxic frame. Core body temperature is maintained with a circulating water blanket, while electrocardiogram, heart rate, oxygen saturation, and temperature are continuously monitored during the procedure. A burr hole is made in the skull with a dental drill to expose an area of ​​dura directly above the target site. AAV2-hAADC is infused into two groups of monkeys, one group (1) through a cannula assembly of the present invention (e.g., cannula assembly 10 shown in Figures 1A-1C) and the other group (2) through a reference cannula assembly that includes a cannula and a needle that is interference-fitted into the opposing ends of the rubber tubing. The rubber tubing in the reference cannula assembly attempts to bridge the space between the cannula and the needle. The reference cannula assembly lacks the hub and polymeric support material included in the cannula assemblies of the present invention.

[0128] Each monkey received a total of 3 x 10 mAbs in 200 µL delivered across four sites. 11 vg (50 μL per site, in two sites per hemisphere). An infusion cannula assembly is manually guided into the putamen in each hemisphere and animals receive AAV2-hAADC (1.5 × 10) at infusion rates of 0.2 μL / min (10 min), 0.5 μL / min (10 min), 0.8 μL / min (10 min), and 1 μL / min (35 min) for the left hemisphere and a constant rate of 1 μL / min (50 min) for the right hemisphere. 12 Each monkey receives a bilateral infusion (i.e., sequential infusion to rostral and caudal sites in both hemispheres) of 1000 mg / mL (vg / mL). Approximately 10 minutes after infusion, the cannula assembly is removed, the wound site is closed, and the monkey is monitored for recovery from anesthesia and then returned to its home cage for continued observation.

[0129] A solution containing gadolinium contrast agent is used in addition to the adeno-associated viral vector carrying the aromatic L-amino acid decarboxylase gene (AAV2-AADC) to track the delivery of the viral vector solution. Visualization of gadolinium indicates that the infused bolus administration device tracks the movement of the cannula assembly tip and closely mimics the shape of the target anatomical structure. However, it is expected that there will be a significant difference in gadolinium intensity in the two groups of monkeys. It is expected that monkeys receiving AAV2-hAADC-gadolinium using the cannula assembly of the present invention will show a significantly stronger gadolinium intensity compared to that in the reference group. Assuming that equal volumes and amounts of viral vector and gadolinium are administered to both groups, the use of the reference cannula assembly would be expected to clearly show leakage of the solution, while the cannula assembly of the present invention will be protected from leakage.

[0130] (Histology and Immunohistochemistry)

[0131] For histological studies, animals are perfused via intracardiac saline infusion followed by 10% neutral buffered formalin (NBF). Brains are then removed and sliced ​​into coronal blocks (8-10 mm) in brain molds. Harvested brain blocks are fixed by immersion in 10% NBF fixative. Tissue blocks are transferred after 2-3 days of fixation into ascending concentrations of PBS / sucrose solutions (10, 20, and 30%) for 3-5 days. Brains are frozen in a bath of isopentane, cooled on dry ice, and cut serially into 40 μm-thick coronal sections on a cryostat. Every 10th section is stained with Hematoxylin and Eosin (H&E) solution (Richard Allen Scientific, Kalamazoo, Mich.) for histopathological analysis. Immunohistochemistry is performed on free-floating sections with a primary antibody specific for AADC (Chemicon (Temecula, Calif.), 1:1,500). Sections are incubated in 3% hydrogen peroxide for 30 minutes to quench endogenous peroxidase. After blocking for nonspecific binding with 10% normal goat serum, sections are incubated in the primary antibody overnight at room temperature, then with a biotinylated anti-rabbit IgG antibody (Vector Laboratories (Burlingame, Calif.), 1:300) and with streptavidin-conjugated horseradish peroxidase (Vector Laboratories, 1:300), both for 1 hour at room temperature. Complexes are visualized with 3-3'-diaminobenzidine (DAB, Vector Laboratories) and hydrogen peroxide. Sections were mounted on Superfrost Plus® slides (Brain Research Laboratories, Newton, Mass.), dried, dehydrated in an ascending ethanol series, cleared in xylene, and mounted with Cytoseal-XYL (Richard-Allen Scientific, Kalamazoo, Mich.).The anteroposterior distribution of hAADC immunostaining was determined by the formula (n x 10 x 40 μm), where n is the number of sections with hAADC positive cells, 40 μm is the thickness of the section, and every 10th section is investigated. The volume of distribution was recorded using a video camera and a Stereoinvestigator. TM Stereological parameters were estimated in serial sections (every 10th) stained for AADC using a photofractionator-photodissociator design-based stereology method under 63x magnification on a Zeiss microscope equipped with stereology software (Microbrightfield, Williston, Vt.). CEE was <5% for each group. Results are reported as mean ± SD. Student's t-test was used to measure statistical significance.

[0132] (Real-time quantitative PCR)

[0133] The vector AAV2-hAADC used in this study contains human AADC target cDNA. Real-time Q-PCR primers and probes anneal to exons 2 and 3 of the AADC gene and span an intron not present in the vector sequence, thereby minimizing amplification of genomic DNA. Real-time Q-PCR is standardized with linearized plasmid DNA containing the vector insert, and the vector genome is quantified as previously described (Sommer et al. (2003) Mol. Ther. 7(1):122-8).

[0134] Immunohistochemistry and quantification of hAADC expression in vivo

[0135] Immunohistological analysis of hAADC expression is performed on each hemisphere 5.5 weeks after AAV-hAADC-2 infusion to determine whether vector distribution differs between infusions with the cannula assembly of the present invention and the reference cannula assembly. All monkeys show hAADC expression in the putamen. Sequential sections were examined for hAADC-positive cells using brightfield microscopy. The volume of distribution and anterior-posterior (AP) divergence of hAADC transgene-positive cells are determined for all animals. The volume of distribution is expected to be significantly different in the two groups of monkeys, i.e., monkeys infused with the cannula assembly of the present invention are expected to be significantly higher than the other group, and this difference is due to the expected leakage of the viral vector solution in the reference cannula assembly compared to the no leakage condition using the cannula assembly of the present invention.

[0136] In all animals, transgene expression is localized to the putamen. No hAADC expression is detected in cortical regions except in areas in line with the infusion trajectory. No differences are observed in the number of AADC-positive cells or the intensity of hAADC staining in comparison of the right and left hemispheres in each group of monkeys. However, in the group in which delivery is performed using the reference cannula assembly, significantly less hAADC staining is expected to be observed by observation with gadolinium contrast at the time of delivering the viral vector. In other words, the reference cannula assembly is expected to show significant leakage compared to the cannula assembly of the present invention, which protects against leakage. The observed lower expression of hAADC in the group in which the reference cannula assembly is used compared to the group with the cannula assembly of the present invention is therefore due to the significantly smaller volume, and therefore less titer, of viral vector infused due to leakage during delivery, with all other experimental conditions between these two groups being otherwise the same.

[0137] Quantitative recovery of vectors is assessed through simulated infusions using the cannula assembly of the present invention and a reference cannula assembly. In both cases, AAV2-hAADC vectors were administered at 5×10 11 vg / mL (0.5 × 10 9 The vector is diluted to 100 µg / µL (100 µL / mL). After filling, both cannula assemblies are pumped with 500 µL of vector solution at 8 µL / min (62.5 min). Two sequential aliquots of 50 µL are collected from the triplicate set of each cannula assembly at flow rates of 0.2-1.0 µL / min. The vector concentration in each sample is determined by real-time quantitative PCR (Q-PCR). While the recovery for the reference cannula assembly is only 49 ± 15% after 1 h of extensive pumping, full recovery of vector (101 ± 3%) is observed for the cannula assembly of the present invention. This full vector recovery further confirms the superior protection from leakage within the cannula assembly of the present invention.

[0138] The experiments described herein utilized animals with pre-existing NAb titers ranging from 1:1 to 1:100, excluding neutralizing antibodies as a confounding variable, and these titers have no apparent effect on hAADC expression in the putamen. Moreover, post-infusion titers increased only slightly after vector administration, thereby permitting well-targeted and minimally disruptive gene delivery using the present device and infusion conditions. These results also suggest that repeated intrastriatal infusions of AAV2 may be feasible in human patients.

[0139] In summary, infusion of AAV2-hAADC into monkey putamen via the cannula assembly of the present invention is expected to be protected from leakage and is well tolerated.

[0140] Certain embodiments of the cannula assembly of the present invention are tested to assess their ability to effectively deliver rAAV vectors to the primate brain, which can serve as a model for the delivery of therapeutic rAAV vectors for the treatment of CNS diseases and disorders in human subjects.

[0141] While the disclosed embodiments have been illustrated and described with respect to one or more implementations, equivalent alterations and modifications will occur to or become known to others skilled in the art upon reading and understanding of this specification and the annexed drawings. In addition, while a particular feature of the invention may be disclosed with respect to only one of several implementations, such feature may be combined with one or more other features of other implementations, as may be desirable or advantageous for any given or particular application.

[0142] While various embodiments of the present invention have been described above, it should be understood that they are presented by way of example only, and not by way of limitation. Numerous modifications to the disclosed embodiments can be made in accordance with the disclosures herein without departing from the spirit or scope of the present invention. Thus, the scope and scope of the present invention should not be limited by any of the embodiments described above. Rather, the scope of the present invention should be defined according to the following claims and their equivalents.

Claims

1. 1. A cannula assembly comprising: a cannula having a proximal end and a distal end; a polymeric support material substantially surrounding a portion of the cannula at or near the proximal end; a hub configured to be attached to a syringe; Equipped with the polymeric support material is disposed between the cannula and the hub; A cannula assembly, wherein the cannula, the polymeric support material, and the hub are attached by adhesive.

2. The cannula assembly of claim 1 , wherein the cannula comprises fiberglass.

3. The cannula assembly of claim 1 , wherein the polymeric support material is tapered.

4. The cannula assembly of claim 3 , wherein the polymeric support material tapers from the proximal end to the distal end, the thickness of the polymeric support material being greater at the proximal end.

5. 10. The cannula assembly of claim 1, wherein the polymeric support material comprises polytetrafluoroethylene (PTFE), polyamide, fluoropolymer, polyolefin, PVC (polyvinyl chloride), polyimide, PEEK (polyetheretherketone), or combinations thereof.

6. The cannula assembly of claim 1 , wherein the polymeric support material is polymeric support tubing.

7. The cannula assembly of claim 1 , wherein the polymeric support material completely surrounds the cannula.

8. The cannula assembly of claim 1 , wherein the hub is a Luer interference fit hub.

9. The cannula assembly of claim 1 , wherein the polymeric support material and the hub comprise transparent or translucent materials.

10. A cannula assembly as described in claim 1, wherein the adhesive is a UV-sensitive adhesive.

11. A combination of a cannula assembly and a syringe, said combination comprising: a cannula having a proximal end and a distal end, a polymeric support material substantially surrounding a portion of the cannula at or near the proximal end, the polymeric support material being disposed between the cannula and a hub, the polymeric support material and the hub being attached by an adhesive; the syringe including a needle; Equipped with The hub attaches the cannula and the syringe, forming a cannula assembly and syringe combination.

12. The combination of claim 11 , wherein the polymeric support material is tapered.

13. 13. The combination of claim 12, wherein the polymeric support material tapers from the proximal end to the distal end, the thickness of the polymeric support material being greater at the proximal end.

14. The combination of claim 11 , wherein the polymeric support material completely surrounds the cannula.

15. 1. A method for forming a cannula assembly, the method comprising: providing a cannula having a proximal end and a distal end, a polymeric support material, and a hub, the polymeric support material and the hub comprising a transparent or translucent material; disposing the polymeric support material to substantially surround a portion of the cannula at or near the proximal end, the polymeric support material being disposed between the cannula and the hub; placing an adhesive on at least one of the cannula, the polymeric support material, and the hub; exposing the adhesive to ultraviolet light to securely attach the cannula, the polymeric support material, and the hub; A method comprising:

16. The method of claim 15 , wherein the polymeric support material is tapered.

17. 17. The method of claim 16, wherein the polymeric support material tapers from the proximal end to the distal end, the thickness of the polymeric support material being greater at the proximal end.

18. The method of claim 15 , wherein the polymeric support material completely surrounds the cannula.

19. 1. A method for forming a cannula assembly, the method comprising: providing a cannula having a proximal end and a distal end, a polymeric support material, and a hub; disposing a polymeric support material to substantially surround a portion of the cannula at or near the proximal end, the polymeric support material being disposed between the cannula and the hub; shrink wrapping the hub and the polymeric support material about the cannula to securely attach the cannula, the polymeric support material, and the hub; A method comprising:

20. The combination described in claim 11, wherein the combination is configured to deliver a viral vector to the central nervous system of a subject.

21. The combination of claim 20 , wherein the viral vector is a recombinant viral vector.

22. The combination of claim 20, wherein the viral vector is in a dosage of about 0.5E9 to about 1.5E9 vg / μl.

23. The combination of claim 22, wherein the viral vector is in a dosage of about 0.7E9 to about 1.3E9 vg / μl.

24. 21. The combination of claim 20, wherein the central nervous system is brain tissue.

25. 21. The combination of claim 20, wherein the central nervous system is the spinal cord.

26. 21. The combination of claim 20, wherein the subject has a neurological disorder.

27. 27. The combination of claim 26, wherein the neurological disorder is meningitis, encephalitis, multiple sclerosis (MS), stroke, brain tumor, epilepsy, Alzheimer's disease, AIDS dementia, Parkinson's disease, or Huntington's disease.

28. 28. The combination of claim 27, wherein the neurological disorder is Alzheimer's disease, Parkinson's disease, or Huntington's disease.

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