Platform attachment for angled subcutaneous injection and methods of use thereof

Abstract

In one aspect, the present disclosure provides a platform attachment for angled subcutaneous injection and for delivery of a medicament comprising hyaluronidase. In another aspect, the present disclosure provides a method of administering a high volume of the formulation in a single administration to treat a disease or disorder in a subject via a platform attachment for angled subcutaneous injection.

Description

TITLEPlatform Attachment for Angled Subcutaneous Injection and Methods of Use ThereofTECHNICAL FIELD

[0001] The present disclosure generally relates to a platform attachment for angled subcutaneous injection and methods of use thereof, e g., administration of pharmaceutical formulations comprising a hyaluronidase enzyme.BACKGROUND

[0002] Current auto-injectors (AIs) are limited to extremely small volumes, limiting their usefulness for delivery of larger volumes. Current biologies typically can have dose volumes of 5-50 mL. Higher flow rates are required to make use of an Al practical for doses of larger volumes.SUMMARY OF THE DISCLOSURE

[0003] The disclosure provides a method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject via subcutaneous administration about 3 mL to about 50 mL of a formulation comprising a therapeutically effective amount of an active ingredient selected from a small molecule, a peptide fragment, a biologic, a nanoparticle, an antibody, an antibody fragment, and a small molecule antiviral; wherein the subcutaneous administration is facilitated by a subcutaneous administration system comprising a platform attachment comprising a base having a generally flat distal surface configured to contact a subject’s skin, the base having an aperture extending therethrough and a housing configured to receive and support an injection apparatus containing the formulation, the housing extending non- perpendicularly in a proximal direction from the base, the housing forming a conduit in fluid communication with the aperture of the base.

[0004] In some embodiments, the injection apparatus containing the formulation further comprises a needle extending into the housing and through the aperture of the base during use. In some embodiments, the housing comprises indicia to indicate a preferred alignment of the injection apparatus relative to the housing. In some embodiments, the injection apparatus is coupled to the housing in the preferred alignment, an open portion of the needle is facing generally perpendicular to the subject’s skin. In some embodiments, the injection apparatus containing the formulation is an auto-injector. In some embodiments, the auto-injector contains at least 3 mL of the formulation. Insome embodiments, the injection apparatus containing the formulation is an injection line. In some embodiments, the distal surface of the base is a generally reuleaux triangle shape. In some embodiments, the distal surface of the base comprises a lip extending around a periphery and protruding distally therefrom. In some embodiments, the distal surface of the base comprises one or more ridges protruding distally therefrom. In some embodiments, an internal surface of the housing comprises at least one protrusion extending radially thereof.

[0005] In some embodiments, the housing extends at an angle of about 5 degrees, about 10 degrees, about 15 degrees, about 20 degrees, about 25 degrees, about 30 degrees, about 35 degrees, about 40 degrees, about 45 degrees, about 50 degrees, about 55 degrees, about 60 degrees, about 65 degrees, about 70 degrees, about 75 degrees, about 80 degrees, about 85 degrees, or less than about 90 degrees relative to the distal surface of the base. In particular embodiments, the housing extends at an angle of 45 degrees relative to the distal surface of the base. In some embodiments, the conduit of the housing includes threads configured to engage corresponding threads of the injection apparatus.

[0006] In some embodiments, the method comprises administering to the subject via subcutaneous administration about 3 mL to about 50 mb of a formulation comprising a therapeutically effective amount of an active ingredient selected from a small molecule, a peptide fragment, a biologic, a nanoparticle, an antibody, an antibody fragment, and a small molecule antiviral; wherein the subcutaneous administration is facilitated by a subcutaneous administration system comprising a platform attachment comprising a base having a generally flat distal surface configured to contact a subject’s skin, the base having an aperture extending therethrough; and a housing configured to receive and support an injection apparatus containing the formulation, the housing extending non-perpendicularly in a proximal direction from the base, the housing forming a conduit in fluid communication with the aperture of the base, wherein the injection apparatus containing the formulation comprises a needle extending into the housing and through the aperture of the base during use.

[0007] In some embodiments, the base is a generally reuleaux triangle shape. In some embodiments, the method comprises attaching to a subject’s skin a platform attachment having a housing with an aperture extending therethrough and a generally flat base angled relative to the housing; and attaching to the housing an injection apparatus containing a formulation such that a needle extends distally through the aperture and into the subject’s skin.

[0008] In some embodiments, attaching the injection apparatus to the housing inserts the needle subcutaneously in the subject’s skin at an angle of about 5 degrees, about 10 degrees, about 15degrees, about 20 degrees, about 25 degrees, about 30 degrees , about 35 degrees, about 40 degrees, about 45 degrees, about 50 degrees, about 55 degrees, about 60 degrees, about 65 degrees, about 70 degrees, about 75 degrees, about 80 degrees, about 85 degrees, or less than about 90 degrees relative to the base. In some embodiments, the method further comprises delivering about 3 mL to about 50 mL of the formulation through the needle. In some embodiments, attaching the injection apparatus to the housing comprises attaching an auto-injector to the housing. In some embodiments, attaching the injection apparatus to the housing comprises attaching an injection line to the housing.

[0009] In some embodiments, the formulation further comprises a hyaluronidase enzyme. In some embodiments, the hyaluronidase enzyme is a recombinant human hyaluronidase enzyme. In some embodiments, the hyaluronidase enzyme is recombinant human hyaluronidase PH20 enzyme. In some embodiments, the hyaluronidase enzyme has an activity of about 150 U / mL to about 150 kU / mL. In some embodiments, the hyaluronidase enzyme has an activity of about 500 U / mL to about 5,000 U / mL. In some embodiments, the hyaluronidase enzyme has an activity of about 1,500 U / mL to about 10,000 U / mL. In some embodiments, the active ingredient is a small molecule, a peptide fragment, a biologic, or a nanoparticle. In some embodiments, the active ingredient is an antibody, an antibody fragment, or a small molecule antiviral. In some embodiments, the method comprises administering to the subject about 10 mL to about 20 mL of the formulation. In some embodiments, the method comprises administering to the subject about 3 mL to about 25 mL of the formulation.

[0010] In some embodiments, the method comprises administering to the subject about 3 mL, about 3.1 mL, about 3.2 mL, about 3.4 mL, about 3.5 mL, about 3.6 mL, about 3.7 mL, about 3.8 mL, about 3.9 mL, about 4 mL, about 4.1 mL, about 4.2 mL, about 4.3 mL, about 4.4 mL, about 4.5 mL, about 5 mL, about 5.1 mL, about 5.2 mL, about 5.3 mL, about 5.4 mL, about 5.5 mL, about 5.6 mL, about 5.7 mL, about 5.8 mL, about 5.9 mL, about 6 mL, about 6.1 mL, about 6.2 mL, about 6.3 mL, about 6.4 mL, about 6.5 mL, about 6.6 mL, about 6.7 mL, about 6.8 mL, about 6.9 mL, about 7 mL, about 7.1 mL, about 7.2 mL, about 7.3 mL about 7.4 mL, about 7.5 mL, about 7.6 mL, about 7.7 mL, about 7.8 mL, about 7.9 mL, about 8 mL, about 8.1 mL, about 8.2 mL, about 8.3 mL, about8.4 mL, about 8.5 mL, about 8.6 mL, about 8.7 mL, about 8.8 mL, about 8.9 mL, about 9 mL, about 9.1 mL, about 9.2 mL, about 9.3 mL, about 9.4 mL, about 9.5 mL, about 9.6 mL, about 9.7 mL, about 9.8 mL, about 9.9 mL, about 10 mL, about 10.1 mL, about 10.2 mL, about 10.3 mL, about10.4 mL, about 10.5 mL, about 10.6 mL, about 10.7 mL, about 10.8 mL, about 10.9 mL, about 11 mL, about 11.1 mL, about 11.2 mL, about 11.3 mL, about 11.4 mL, about 11.5 mL, about 11.6 mL, about 11.7 mL, about 11.8 mL, about 11.9 mL, about 12 mL, about 12.1 mL, about 12.2 mL, about12.3 mL, about 12.4 m , about 12.5 mb, about 12.6 mb, about 12.7 mb, about 12.8 mL, about 12.9 mb, about 13 mL, about 13.1 mL, about 13.2 mL, about 13.3 mL, about 13.4 mL, about 13.5 mL, about 13.6 mL, about 13.7 mL, about 13.8 mL, about 13.9 mL, about 14 mL, about 14.1 mL, about14.2 mL, about 14.3 mL, about 14.4 mL, about 14.5 mL, about 14.6 mL, about 14.7 mL, about 14.8 mL, about 14.9 mL, about 15 mL, about 15.1 mL, about 15.2 mL, about 15.3 mL, about 15.4 mL, about 15.5 mL, about 15.6 mL, about 15.7 mL, about 15.8 mL, about 15.9 mL, about 16 mL, about 16.1 mL, about 16.2 mL, about 16.3 mL, about 16.4 mL, about 16.5 mL, about 16.6 mL, about 16.7 mL, about 16.8 mL, about 16.9 mL about 17 mL, about 17.1 mL, about 17.2 mL, about 17.3 mL, about 17.4 mL, about 17.5 mL, about 17.6 mL, about 17.7 mL, about 17.8 mL, about 17.9 mL, about 18 mL, about 18.1 mL, about 18.2 mL, about 18.3 mL, about 18.4 mL, about 18.5 mL, about 18.6 mL, about 18.7 mL, about 18.8 mL, about 18.9 mL, about 19 mL, about 19.1 mL, about 19.2 mL, about 19.3 mL, about 19.4 mL, about 19.5 mL, about 19.6 mL, about 19.7 mL, about 19.8 mL, about 19.9 mL, about 20 mL, about 20.1 mL, about 20.2 mL, about 20.3 mL, about 20.4 mL, about 20.5 mL, about 20.6 mL, about 20.7 mL, about 20.8 mL, about 20.9 mL, about 21 mL, about 21.1 mL, about 21.2 mL, about 21.3 mL, about 21.4 mL, about 21.5 mL, about 21.6 mL, about 21.7 mL, about 21.8 mL, about 21.9 mL, about 22 mL, about 22.1 mL, about 22.2 mL, about 22.3 mL, about22.4 mL, about 22.5 mL, about 22.6 mL, about 22.7 mL, about 22.8 mL, about 22.9 mL, about 23 mL, about 23.1 mL, about 23.2 mL, about 23.3 mL, about 23.4 mL, about 23.5 mL, about 23.6 mL, about 23.7 mL, about 23.8 mL, about 23.9 mL, about 24 mL, about 24.1 mL, about 24.2 mL, about24.3 mL, about 24.4 mL, about 24.5 mL, about 24.6 mL, about 24.7 mL, about 24.8 mL, about 24.9 mL, or about 25 mL.

[0011] In some embodiments, the method comprises administering the formulation using a high volume autoinjector. In some embodiments, the method comprises administering the formulation with a starting delivery force of about 5 Ibf to about 50 Ibf using a high volume autoinjector. In some embodiments, the method comprises administering the formulation with an ending delivery force of about 3 Ibf to about 20 Ibf using a high volume autoinjector. In some embodiments, the method comprises administering the formulation with a starting pressure of about 50 psi to about 200 psi using a high volume autoinjector. In some embodiments, the method comprises administering the formulation with an ending pressure of about 20 psi to about 75 psi using a high volume autoinjector.

[0012] In some embodiments, the formulation is in a prefilled syringe. In some embodiments, the prefilled syringe contains about 3 mL, about 3.1 mL, about 3.2 mL, about 3.4 mL, about 3.5 mL, about 3.6 mL, about 3.7 mL, about 3.8 mL, about 3.9 mL, about 4 mL, about 4.1 mL, about 4.2 mL,about 4.3 mL, about 4.4 mL, about 4.5 mL, about 5 m , about 5.1 mL, about 5.2 mL, about 5.3 mL, about 5.4 mL, about 5.5 mL, about 5.6 mL, about 5.7 mL, about 5.8 mL, about 5.9 mL, about 6 mL, about 6.1 mL, about 6.2 mL, about 6.3 mL, about 6.4 mL, about 6.5 mL, about 6.6 mL, about 6.7 mL, about 6.8 mL, about 6.9 mL, about 7 mL, about 7.1 mL, about 7.2 mL, about 7.3 mL about 7.4 mL, about 7.5 mL, about 7.6 mL, about 7.7 mL, about 7.8 mL, about 7.9 mL, about 8 mL, about 8.1 mL, about 8.2 mL, about 8.3 mL, about 8.4 mL, about 8.5 mL, about 8.6 mL, about 8.7 mL, about 8.8 mL, about 8.9 mL, about 9 mL, about 9.1 mL, about 9.2 mL, about 9.3 mL, about 9.4 mL, about 9.5 mL, about 9.6 mL, about 9.7 mL, about 9.8 mL, about 9.9 mL, about 10 mL, about 10. 1 mL, about 10.2 mL, about 10.3 mL, about 10.4 mL, about 10.5 mL, about 10.6 mL, about 10.7 mL, about 10.8 mL, about 10.9 mL, about 11 mL, about 11.1 mL, about 11.2 mL, about 11.3 mL, about11.4 mL, about 11.5 mL, about 11.6 mL, about 11.7 mL, about 11.8 mL, about 11.9 mL, about 12 mL, about 12.1 mL, about 12.2 mL, about 12.3 mL, about 12.4 mL, about 12.5 mL, about 12.6 mL, about 12.7 mL, about 12.8 mL, about 12.9 mL, about 13 mL, about 13.1 mL, about 13.2 mL, about13.3 mL, about 13.4 mL, about 13.5 mL, about 13.6 mL, about 13.7 mL, about 13.8 mL, about 13.9 mL, about 14 mL, about 14.1 mL, about 14.2 mL, about 14.3 mL, about 14.4 mL, about 14.5 mL, about 14.6 mL, about 14.7 mL, about 14.8 mL, about 14.9 mL, about 15 mL, about 15.1 mL, about 15.2 mL, about 15.3 mL, about 15.4 mL, about 15.5 mL, about 15.6 mL, about 15.7 mL, about 15.8 mL, about 15.9 mL, about 16 mL, about 16.1 mL, about 16.2 mL, about 16.3 mL, about 16.4 mL, about 16.5 mL, about 16.6 mL, about 16.7 mL, about 16.8 mL, about 16.9 mL about 17 mL, about 17.1 mL, about 17.2 mL, about 17.3 mL, about 17.4 mL, about 17.5 mL, about 17.6 mL, about 17.7 mL, about 17.8 mL, about 17.9 mL, about 18 mL, about 18.1 mL, about 18.2 mL, about 18.3 mL, about 18.4 mL, about 18.5 mL, about 18.6 mL, about 18.7 mL, about 18.8 mL, about 18.9 mL, about 19 mL, about 19.1 mL, about 19.2 mL, about 19.3 mL, about 19.4 mL, about 19.5 mL, about 19.6 mL, about 19.7 mL, about 19.8 mL, about 19.9 mL, about 20 mL, about 20.1 mL, about 20.2 mL, about 20.3 mL, about 20.4 mL, about 20.5 mL, about 20.6 mL, about 20.7 mL, about 20.8 mL, about 20.9 mL, about 21 mL, about 21.1 mL, about 21.2 mL, about 21.3 mL, about 21.4 mL, about21.5 mL, about 21.6 mL, about 21.7 mL, about 21.8 mL, about 21.9 mL, about 22 mL, about 22.1 mL, about 22.2 mL, about 22.3 mL, about 22.4 mL, about 22.5 mL, about 22.6 mL, about 22.7 mL, about 22.8 mL, about 22.9 mL, about 23 mL, about 23.1 mL, about 23.2 mL, about 23.3 mL, about23.4 mL, about 23.5 mL, about 23.6 mL, about 23.7 mL, about 23.8 mL, about 23.9 mL, about 24 mL, about 24.1 mL, about 24.2 mL, about 24.3 mL, about 24.4 mL, about 24.5 mL, about 24.6 mL, about 24.7 mL, about 24.8 mL, about 24.9 mL, or about 25 mL of the formulation. In some embodiments, the prefilled syringe comprises a needle having a gauge of about 20 to about 33. Insome embodiments, the prefilled syringe comprises a 20 gauge needle, a 21 gauge needle, a 22 gauge needle, a 23 gauge needle, a 24 gauge needle, a 25 gauge needle, a 26 gauge needle, a 27 gauge needle, a 28 gauge needle, a 29 gauge needle, a 30 gauge needle, a 31 gauge needle, a 32 gauge needle, or a 33 gauge needle.

[0013] In some embodiments, the method comprises administering the formulation at a rate of about 0.08 to about 1.00 mL / sec. In some embodiments, the method comprises administering the formulation at a rate of at least about 0.08 to about 1.0 mL / sec. In some embodiments, the method comprises administering the formulation at a rate of at least or faster than about 0.08 to about 1.00 mL / sec. In some embodiments, the administration takes about 10 seconds to about 40 seconds. In some embodiments, the administration takes at least about 10 seconds to about 40 seconds. In some embodiments, the administration takes at least or less than about 10 seconds to about 40 seconds. In some embodiments, the administration takes about 15 seconds to about 30 seconds. In some embodiments, the administration takes at least about 15 seconds to about 30 seconds. In some embodiments, the administration takes at least or less than about 15 seconds to about 30 seconds.

[0014] In some embodiments, the method comprises administering about 5 mL of the formulation at a rate of about 0.14 mL / sec to about 0.21 mL / sec. In some embodiments, the method comprises administering about 10 mL of the formulation at a rate of about 0.32 mL / sec to about 0.42 mL / sec. In some embodiments, the formulation has a viscosity of about 1 cP to about 50 cP. In some embodiments, administration of the formulation requires less applied force when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments, the method comprises administering about 5 mL of the formulation at a rate of about 0.14 mL / sec to about 0.21 mL / sec with an applied force of about 10 N to about 45 N. In some embodiments, the method comprises administering the formulation to the subject using a prefilled syringe comprising a 25 gauge needle. In some embodiments, the method comprises administering about 10 mL of the formulation to the subject at a rate of about 0.32 mL / sec to about 0.42 mL / sec with an applied force of about 25 N to about 50 N. In some embodiments, the method comprises administering the formulation to the subject using a prefilled syringe comprising a 25 gauge needle.

[0015] In some embodiments, administration of the formulation is faster when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments, administration of the formulation causes fewer side effects in the subject when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments, administration of the formulation causes less pain and discomfort in the subject when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments,administration of the formulation causes less back leakage at the injection site when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments, the back leakage at the injection site is about 85% to about 30% less when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments, administration of the formulation causes less swelling volume and / or swelling height at the injection site when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments, the formulation causes about 35% to about 5% less swelling and / or swelling height the injection site when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments, administration of the formulation yields a lower bleb swelling size, less bleb induration, and / or quicker bleb resolution when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments, administration of the formulation yields more consistent delivery times when compared to a similar formulation that does not comprise a hyaluronidase enzyme. In some embodiments, the subject is human. In some embodiments, the administering comprises the subject self-administering the formulation. In some embodiments, the administering comprises a healthcare provider or a caregiver administering the formulation to the subject. In some embodiments, the subcutaneous administration comprises a single injection. In some embodiments, the subcutaneous administration comprises two or more injections.BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The following detailed description of embodiments of the platform attachment for angled subcutaneous injection, will be better understood when read in conjunction with the appended drawings of exemplary embodiments. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities shown.

[0017] In the drawings:

[0018] Fig. 1A is a perspective view of a platform attachment for angled subcutaneous injection in accordance with an exemplary embodiment of the present invention. Fig. IB is a side view of the platform attachment for angled subcutaneous injection of Fig. 1A.

[0019] Fig. 2 is a side view of the platform attachment for angled subcutaneous injection of Fig. 1A aligned with an auto-injector.

[0020] Fig. 3 is a side view of the platform attachment for angled subcutaneous injection of Fig. 1A coupled to the auto-injector.

[0021] Fig 4 is a side view of the platform attachment of Fig. 1A shown coupled to an injection line.

[0022] Fig. 5A is a top view of the platform attachment for angled subcutaneous injection of Fig. 1A. Fig. 5B is a bottom view of the platform attachment for angled subcutaneous injection of Fig. 1A.

[0023] Fig. 6A is a top view of a slotted handle of the platform attachment for angled subcutaneous injection of Fig. 1A. Fig. 6B is a cross sectional view of a slotted handle of the platform attachment for angled subcutaneous injection of Fig. 1 A. Fig. 6C is a side view of a slotted handle of the platform attachment for angled subcutaneous injection of Fig. 1A.

[0024] Fig. 7 is a bottom view of the platform attachment for angled subcutaneous injection in accordance with a second exemplary embodiment of the present invention.

[0025] Fig. 8 is a graph of HVAI Injection Times (mean ± SEM).

[0026] Fig. 9 is a graph of mean (mg ± SEM) and individual weights of back-leakage.

[0027] Fig. 10 is a graph of individual swelling volumes (mL) after SC injection of Ig-120 and Ig- 120 + rHuPH20 taken via caliper measurement at TO.

[0028] Fig. 11 is a graph of individual swelling area (cm2) after SC injection of Ig-120 and Ig-120 + rHuPH20 taken via caliper measurement at TO.

[0029] Fig. 12 is a graph of individual swelling height (mm) after SC injection of Ig-120 and Ig- 120 + rHuPH20 taken via caliper measurement at TO.

[0030] Fig. 13 is a graph of assessments of swelling volume over time (TO-T15-T3O) taken via caliper measurement (mL ± SEM).

[0031] Fig. 14 is a graph of assessment of swelling area overtime (T0-T15-T30) taken via caliper measurement (cm2± SEM).

[0032] Fig. 15 is a graph of assessment of swelling height overtime (T0-T15-T30 taken via caliper measurement (mm ± SEM).

[0033] Fig. 16 shows images of colorimetric assessments of bleb shape and size using 3D imaging.

[0034] Fig. 17 is a graph of individual bleb volume (mL) after SC injection of Ig-120 and Ig-120 + rHuPH20 taken via 3D imaging.

[0035] Fig. 18 is a graph of individual bleb area (cm2) after SC injection of Ig-120 and Ig-120 + rHuPH20 taken via 3D imaging.

[0036] Fig. 19 is a graph of individual bleb height (mm) after SC injection of Ig-120 and Ig-120 + rHuPH20 taken via 3D imaging.

[0037] Fig. 20 is a graph showing qualitative assessment of post-injection erythema (T0-T15-T30- T2h).

[0038] Fig. 21 is a graph showing qualitative assessment of post-injection swelling size (TO).

[0039] Fig. 22 is a graph showing qualitative assessment of post-injection swelling size (T0-T15-T30-T2h).

[0040] Fig. 23 is a graph showing qualitative assessment of post-injection induration (TO).

[0041] Fig. 24 is a graph showing qualitative assessment of post-injection induration (T0-T15- T30-T2h).

[0042] Fig. 25 shows images of injection site on AID #531 IL: Ig-120 (pre-inj ection to 24h).

[0043] Fig. 26 shows images of injection site on AID #5311R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0044] Fig. 27 shows images of injection site on AID #5312R: Ig-120 (pre-inj ection to 24h).

[0045] Fig. 28 shows images of injection site on AID #5312L: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0046] Fig. 29 shows images of injection site on AID #5313L: Ig-120 (pre-inj ection to 24h).

[0047] Fig. 30 shows images of injection site on AID #5313R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0048] Fig. 31 shows images of injection site on AID #5315R: Ig-120 (pre-inj ection to 24h).

[0049] Fig. 32 shows images of injection site on AID #5315L: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0050] Fig. 33 shows images of injection site on AID #5317L: Ig-120 (pre-inj ection to 24h).

[0051] Fig. 34 shows images of injection site on AID #5317R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0052] Fig. 35 shows images of injection site on AID #5318R: Ig-120 (pre-inj ection to 24h).

[0053] Fig. 36 shows images of injection site on AID #5318L: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0054] Fig. 37 is a graph of applied force (N) during injection (mean ± SEM) of Ig-120 and Ig- 120 + rHuPH20.

[0055] Fig. 38 is a graph of individual applied force (N) during injection of Ig-120 and Ig-120 + rHuPH20.

[0056] Fig. 39 is a graph of mean (mg ± SEM) and individual weights of back-leakage.

[0057] Fig. 40 is a graph of individual swelling volumes (cm3) after SC injection of Ig-120 andIg-120 + rHuPH20 taken via caliper measurement.

[0058] Fig. 41 is a graph of individual swelling areas (cm2) after SC injection of Ig-120 and Ig- 120 + rHuPH20 taken via caliper measurement.

[0059] Fig. 42 is a graph of individual swelling heights (mm) after SC injection of Ig-120 and Ig- 120 + rHuPH20 taken via caliper measurement.

[0060] Fig. 43 show a composite of 3D images by treatment.

[0061] Fig. 44 is a graph of individual bleb volumes (cm3) after SC injection of Ig-120 and Ig-120 + rHuPH20 taken via 3D imaging.

[0062] Fig. 45 is a graph of individual bleb areas (cm2) after SC injection of Ig-120 and Ig-120 + rHuPH20 measured via 3D imaging.

[0063] Fig. 46 is a graph of individual bleb heights (mm) after SC injection of Ig-120 and Ig-120 + rHuPH20 measured via 3D imaging.

[0064] Fig. 47 is a graph of change in surface temperature: pre to post-injection.

[0065] Fig. 48 is a graph of qualitative assessment of post-injection erythema.

[0066] Fig. 49 is a graph of qualitative assessment of post-injection swelling size.

[0067] Fig. 50 is a graph of qualitative assessment of post-injection induration (firmness).

[0068] Fig. 51 shows images of injection site on AID #1182L: Ig-120 (pre-inj ection to 24h).

[0069] Fig. 52 shows images of injection site on AID #1182R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0070] Fig. 53 shows images of injection site on AID #1183L: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0071] Fig. 54 shows images of injection site on AID #1183R: Ig-120 (pre-inj ection to 24h).

[0072] Fig. 55 shows images of injection site on AID #1 186L: Ig-120 (pre-inj ection to 24h).

[0073] Fig. 56 shows images of injection site on AID #1186R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0074] Fig. 57 shows images of injection site on AID 1187L: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0075] Fig. 58 shows images of injection site on AID #1187R: Ig-120 (pre-inj ection to 24h).

[0076] Fig. 59 shows images of injection site on AID #1189L: Ig-120 (pre-inj ection to 24h).

[0077] Fig. 60 shows images of injection site on AID #1189R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0078] Fig. 61 shows images of injection site on AID 1190L: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0079] Fig. 62 shows images of injection site on AID #1190R: Ig-120 (pre-injection to 24h).

[0080] Fig. 63 is a graph of HVAI injection times (mean ± SEM).

[0081] Fig. 64 is a graph of individual swelling volumes (mL) after SC injection of Ig-120 andIg-120 + rHuPH20 taken via caliper measurement at TO.

[0082] Fig. 65 is a graph of individual swelling area (cm2) after SC injection of Ig-120 and Ig- 120 + rHuPH20 taken via caliper measurement at TO.

[0083] Fig. 66 is a graph of individual swelling height (mm) after SC injection of Ig-120 and Ig- 120 + rHuPH20 taken via caliper measurement at TO.

[0084] Fig. 67 is a graph of assessment of swelling volume over time (T0-T15-T30) taken via caliper measurement (mL ± SEM).

[0085] Fig. 68 is a graph of assessment of swelling area over time (T0-T15-T30) taken via caliper measurement (cm2± SEM).

[0086] Fig. 69 is a graph of assessment of swelling height over time (T0-T15-T30) taken via caliper measurement (mm ± SEM).

[0087] Fig. 70 is an image of colorimetric assessment of bleb shape and size taken via 3D imaging.

[0088] Fig. 71 is a graph of individual bleb volume (mL) after SC injection of Ig-120 and Ig-120 + rHuPH20 taken via 3D imaging.

[0089] Fig. 72 is a graph of individual bleb area (cm2) after SC injection of Ig-120 and Ig-120 + rHuPH20 taken via 3D imaging.

[0090] Fig. 73 is a graph of individual bleb height (mm) after SC injection of Ig-120 and Ig-120 + rHuPH20 taken via 3D imaging.

[0091] Fig. 74 is a graph of qualitative assessment of post-injection erythema at TO.

[0092] Fig. 75 is a graph of qualitative assessment of post-injection erythema over time (T0-T15-T30-T2h).

[0093] Fig. 76 is a graph of qualitative assessment of post-injection swelling size at TO.

[0094] Fig. 77 is a graph of qualitative assessment of post-injection swelling size over time (T0-T15-T30-T2h).

[0095] Fig. 78 is a graph of qualitative assessment of post-injection swelling size over time (T0- T15-T30-T2h-T24h).

[0096] Fig. 79 is a graph of qualitative assessment of post-injection induration at TO.

[0097] Fig. 80 is a graph of qualitative assessment of post-injection induration over time (T0-T15-T30-T2h).

[0098] Fig. 81 is a graph of qualitative assessment of post-injection induration over time (T0- T15-T30-T2h-T24h).

[0099] Fig 82 is a graph of mean injection times at all injection angles (90°-45°-20°).

[0100] Fig. 83 shows a comparison of back-leakage for all injection angles (90°-45°-20°).

[0101] Fig. 84 shows a comparison of post-injection bleb swelling volume at TO.

[0102] Fig. 85 shows a comparison of post-injection swelling volume over time at all injection angles (90°-45°-20°).

[0103] Fig. 86 shows a comparison of post-injection bleb swelling area at TO.

[0104] Fig. 87 shows a comparison of post-injection bleb area (cm2± SEM).

[0105] Fig. 88 shows a comparison of post-injection swelling height at TO.

[0106] Fig. 89 is a graph of measurement of bleb height over time at all injection angles (90°-45°-20°).

[0107] Fig. 90 shows images of injection site on AID #645 IL: Ig-120 (pre-inj ection to 24h).

[0108] Fig. 91 shows images of injection site on AID #6451R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0109] Fig. 92 shows images of injection site on AID #6478R: Ig-120 (pre-inj ection to 24h).

[0110] Fig. 93 shows images of injection site on AID #6478L: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0111] Fig. 94 shows images of injection site on AID #6481L: Ig-120 (pre-inj ection to 24h).

[0112] Fig. 95 shows images of injection site on AID #6481R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0113] Fig. 96 shows images of injection site on AID #6482L: Ig-120 (pre-inj ection to 24h).

[0114] Fig. 97 shows images of injection site on AID #6482R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0115] Fig. 98 shows images of injection site on AID #6484L: Ig-120 (pre-inj ection to 24h).

[0116] Fig. 99 shows images of injection site on AID #6484R: Ig-120 + rHuPH20 (pre-inj ection to 24h).

[0117] Fig. 100 shows images of injection site on 100 AID #6491R: Ig-120 (pre-inj ection to 24h).

[0118] Fig. 101 shows images of injection site on AID #6491L: Ig-120 + rHuPH20 (preinjection to 24h).DETAILED DESCRIPTION

[0119] The present disclosure generally relates to a platform attachment for facilitating angled subcutaneous drug delivery using a device having a needle such as auto-injectors (Al) or injection line. Typically, with Al devices, needles are inserted to a patient’s skin generally at a perpendicular angle relative to the skin surface for subcutaneous or intramuscular injections. Injecting a drug in such an orientation could limit the direction of the outward flow of the medicament via the needle causing swelling under the skin of the patient. When considering large dose volumes, this becomes especially important because the amount of swelling is typically proportional to the dose volume. The greater the dose volume, the larger the post-injection swelling.

[0120] Referring to the drawings in detail, wherein like reference numerals indicate like elements throughout, there is shown in Figs. 1-3 a platform attachment for angled subcutaneous (SC) drug delivery, generally designated 10, in accordance with a first exemplary embodiment of the present invention. The platform attachment is a separate or integral component that aligns and contacts the injection apparatus with the skin surface to control the angle that the needle is inserted into the patient’s subcutaneous or intramuscular tissue.

[0121] Uptake of the medicament into systemic circulation can be limited if the injection is localized. An angled injection may help to spread the medicament within the SC or intramuscular (IM) layer as much as possible during injection. Without sufficient spreading, a bulge (e.g. bleb or wheal) may form around or at the injection site.

[0122] Referring to Figs. 1A-1B, the platform attachment 10 may be comprised of two primary components, a base 12 and a housing 16. The base 12 and the house 16 may be a single continuous unit. The housing 16 may extend from the base 12 at an angle relative to the base. The base 12 may contact a patient’s skin 13 while the housing 16 may receive an injection apparatus 18 for injecting the medicament, as shown in Figs. 2-3.

[0123] The base 12 may have a generally flat distal surface configured to contact a patient’s skin 13. Referring to Figs. 2-4, the skin 13 may have multiple layers. The patient’s skin 13 may be the epidermis layer of the skin. The dermis layer 17 may be the layer immediately below the patient’s skin. The subcutaneous layer 19 may be the layer immediately below the dermis layer 17. The intramuscular layer 23 may be the layer immediately below the subcutaneous layer 19. The dermis layer 17 may be between the skin 13 and the subcutaneous layer 19. The subcutaneous layer 19 may be between the dermis layer 17 and the intramuscular layer 23.

[0124] Referring to Figs. 5A-5B, in order to accurately insert the needle 21, the patient’s skin 13 may be stretched to avoid trampolining or pillowing during insertion. The base 12 may be a generally reuleaux triangle shape. This shape may stretch the skin 13 when the base 12 contacts the patient’sskin 13. This generally reuleaux triangle shape may further provide a directionality that can be used to visualize the direction of the angled needle insertion. The base 12 may be sized with a surface area sufficient to provide a stable placement of the device on the skin. The surface of the base 12 contacting the patient may have a stippled surface to promote surface contact with the skin 13. This textured surface should be orientated so that it is orthogonal to the directionality of the injection. In one embodiment, the base 12 is a generally circular shape. The base 12 may be larger than the housing 16. In one embodiment, the base 12 is the same size as the housing 16. The proximal end of the housing 16 may extend only partially to a perimeter of the base 12. In one embodiment, the housing 16 extends away from a center of the base 12 such that a proximal end of the housing 16 may extend beyond the base 12.

[0125] Each side of the generally reuleaux triangle base 12 may be 1.5 in. long. Each side of the generally reuleaux triangle base 12 may be at least 1.75 in. long. Each side of the generally reuleaux triangle base 12 may be at least 2 in. long. Each side of the generally reuleaux triangle base 12 may be at least 2.25 in. long. Each side of the generally reuleaux triangle base 12 may be at least 2.5 in. long. Each side of the generally reuleaux triangle base 12 may be at least 2.75 in. long. Each side of the generally reuleaux triangle base 12 may be at least 3 in. long. Each side of the generally reuleaux triangle base 12 may be at least 3.25 in. long. Each side of the generally reuleaux triangle base 12 may be at least 3.5 in. long.[00126J The base 12 may have a surface area of at least 1 in2. The base 12 may have a surface area of at least 1.5 in2. The base 12 may have a surface area of at least 2 in2. The base 12 may have a surface area of at least 2.5 in2. The base 12 may have a surface area of at least 3 in2. The base 12 may have a surface area of at least 3.5 in2. The base 12 may have a surface area of at least 4 in2. The base 12 may have a surface area of at least 4.5 in2. The base 12 may have a surface area of at least 5 in2. The base 12 may have a surface area of at least 5.5 in2. The base 12 may have a surface area of at least 6 in2. The base 12 may have a surface area of at least 6.5 in2. The base 12 may have a surface area of at least 7 in2. The base 12 may have a surface area of at least 7.5 in2. The base 12 may have a surface area of at least 8 in2. The base 12 may have a surface area of at least 8.5 in2. The base 12 may have a surface area of at least 9 in2. The base 12 may have a surface area of at least 9.5 in2. The base 12 may have a surface area of at least 10 in2. The base 12 may have a surface area of at least 10.5 in2. The base 12 may have a surface area of at least 11 in2.

[0127] The base 12 may have an aperture 14 (as shown in Fig. 1 A and 5B) extending therethrough. The aperture 14 may be sized to allow a luer. The aperture 14 may be a generally circular shape. In some embodiments, the aperture 14 is oval or rectangular shaped. The aperture 14 may be sized toallow a needle 21 pass therethrough. The aperture 14 may be sized and shaped to allow a cap (not shown) of the needle 21 to pass therethrough and to allow the cap to be held in place until use to minimize needle exposure.

[0128] The aperture may have a diameter of at least 15 mm. The aperture may have a diameter of at least 20 mm. The aperture may have a diameter of at least 25 mm. The aperture may have a diameter of at least 30 mm. The aperture may have a diameter of at least 35 mm. The aperture may have a diameter of at least 40 mm. The aperture may have a diameter of at least 45 mm.

[0129] The platform attachment 10 may have a housing 16 configured to receive and support an injection apparatus 18 (as shown in Figs. 2-3) containing the medicament. The housing 16 may have generally the same shape as the injection apparatus 18. The housing 16 may extend non- perpendicularly in a proximal direction from the base 12. The housing 16 may be sized to contain the injection apparatus 18 and all components necessary for the operation of the injection apparatus 18. The housing 16 may be a generally cylindrical shape. In one embodiment, the housing 16 may be a generally rectangular tube shape. The housing 16 may have a longer length on a proximal side than a distal side depending on the angle A at which it is fixed relative to the base 12. The housing 16 may form a conduit 20 in fluid communication with the aperture 14 of the base 12. The conduit 20 may have a length that is less than a length of the injection apparatus 18 to allow for insertion and removal of the injection apparatus 18. The conduit 20 may have a width that is greater than a width of the injection apparatus 18 to allow the injection apparatus to be received therein. As shown in Fig. 1A, the base 12 may contain an alignment bar 29 on a proximal side. The alignment bar 29 may protrude from the base 12 and be a generally straight line to indicate the location of the aperture 14. In one embodiment, there may be three alignment bars 29 on the base 12 to indicate the location of the aperture 14.

[0130] The housing 16 may have a length of at least 0.25 in. The housing 16 may have a length of at least 0.5 in. The housing 16 may have a length of at least 0.75 in. The housing 16 may have a length of at least 1 in. The housing 16 may have a length of at least 1.25 in. The housing 16 may have a length of at least 1.5 in. The housing 16 may have a length of at least 1.75 in. The housing 16 may have a length of at least 2 in. The housing 16 may have a length of at least 2.25 in. The housing 16 may have a length of at least 2.5 in.

[0131] The injection apparatus 18 containing the medicament may comprise a needle 21 extending into the housing 16 and through the aperture 14 of the base 12 during use (as shown in Fig. 3). The needle 21 may be threadedly coupled to the injection apparatus 18. In one embodiment, the needle 21 may be a staked needle that is pre-attached to the injection apparatus 18. The needle 21 may be alength appropriate to reach the SC layer with a target injection depth of 6-12 mm from the surface of the skin. In some applications, intramuscular administration may be performed using the injection apparatus 18. If the injection apparatus 18 is intended for intramuscular administration of a medicament, a longer needle 21 may be required to reach the underlying intramuscular layer. Because the thickness of the subcutaneous space can vary between individuals, the desired needle length may also vary but a target injection depth of 25-30 mm is desired. Referring to the magnified view in Fig. 3, the needle 21 may have an angled tip at a distal end to pierce the patient’s skin 13. The angled tip of the needle 21 may be a bevel.

[0132] The needle 21 may have a length of at least 7.5 mm. The needle 21 may have a length of at least 10 mm. The needle 21 may have a length of at least 12.5 mm. The needle 21 may have a length of at least 15 mm. The needle 21 may have a length of at least 17.5 mm. The needle 21 may have a length of at least 20 mm. The needle 21 may have a length of at least 22.5 mm. The needle 21 may have a length of at least 25 mm. The needle 21 may have a length of at least 27.5 mm. The needle 21 may have a length of at least 30 mm. The needle 21 may have a length of at least 32.5 mm.

[0133] Referring to Figs. 2-3 the housing 16 may comprise a housing indicator 32 to indicate a preferred orientation of the injection apparatus 18 relative to the housing 16. For example, the indicia may indicate where the bevel of the needle should point to orient as shown in Fig. 3 to optimize spreading of the medicament during use. The housing 16 may include the housing indicator 32 thereon for indicating a center point of the housing 16 and / or a point that the injection apparatus 18 should align to ensure a consistent coupling configuration. The housing indicator 32 may be a graphic of an arrow signifying that a corresponding indicia on the injection apparatus 18 should be aligned therewith. The housing indicator 32 may be a combination of shapes and / or words, such as “align here.” The housing indicator 32 may be engraved in the housing 16. In one embodiment, the housing indicator 32 may be a decal or paint.

[0134] The injection apparatus 18 may also comprise corresponding indicia or apparatus indicator 34 to indicate a preferred alignment of the injection apparatus 18 relative to the housing 16. Apparatus indicator 34 may be provided to indicate a point of the injection apparatus 18 and / or a point that the housing 16 should align with to ensure a preferred coupling configuration. The apparatus indicator 34 may be a graphic of an arrow signifying that a corresponding indicia on the housing 16 should be aligned therewith. The apparatus indicator 34 may be a combination of shapes and / or words, such as “align here.” The apparatus indicator 34 may be engraved in the injection apparatus 18. In one embodiment, the apparatus indicator 34 may be a decal fixed to the injection apparatus 18 with an adhesive.

[0135] When the injection apparatus 18 is coupled to the housing 16 in the preferred alignment, as discussed above, an open portion of the needle 21 may be facing generally parallel and pointing upwards towards the skin surface to facilitate medicament spreading and absorption. The open portion of the needle 21 facing generally parallel to the skin surface may promote the medicament moving into the lymphatic valves and / or systemic capillaries and out of the SC space. The open portion of the needle 21 facing generally parallel to the skin surface may also limit the interaction between the medicament and the muscular layer, which may reduce swelling at the injection site.

[0136] An internal surface 30 of the housing 16 may comprise at least one protrusion 36 (as shown in Fig. 5 A) extending radially thereof to ensure the preferred alignment of the injection apparatus 18 and the housing 16. The protrusions 36 may be a generally triangular shape. In one embodiment, the protrusions 36 are rounded. The protrusion 36 may prevent the injection apparatus 18 from coupling to the housing 16 if the injection apparatus 18 and the housing 16 are not aligned. In one embodiment, there are two protrusions 36. In one embodiment, there are three protrusions 36.

[0137] Apart from the preferred orientation discussed above in connection with the open portion of the needle 21 facing in a generally perpendicular to the skin surface, orientation between the housing 16 and injection apparatus 18 is also important to ensure an angle of the needle 21 relative to the base 12. The angle of the needle 21 relative to the base 12 may be fixed, predetermined, or adjustable. A slotted handle 40 may be coupled to the injection apparatus 18 by a threaded coupling prior to the injection apparatus 18 coupling to the housing 16. Referring to Figs. 6A-6C, the slotted handle 40 may be a generally cylindrical member with a slot 42 extending through one side thereof. The slot 42 may be sized to be wider than a width of a cap of the needle 21 to receive the cap therethrough, and a central hub 44 in a center of the slotted handle 40 shaped to fit around the base of the needle 21.

[0138] The slotted handle 40 may include a threading 46 around an outer periphery of a proximal end thereof to couple the slotted handle 40 to the housing 16. The slotted handle 40 may engage the base of the needle 21 with the central hub 44 may hold the needle in place during use to ensure an angle of the needle 21 relative to the base 12. The slotted handle 40 may engage a stop 48 (as shown in Fig. 2) in the housing 16 for modulating the depth of the needle 21. The slotted handle 40 may be sized to achieve a target depth of the needle 21 in the skin 13. The stop 48 may have threading configured to receive the threading 46 of the slotted handle 40. In one embodiment, the stop 48 may include a snap-fit coupling, or other securing coupling, to fix the stop 48 to the slotted handle 40 thereby fixing the injection apparatus 18 to the platform attachment 10. Such fixing may allow forthe platform attachment 10 and injection apparatus 18 to be moved from the skin 13 simultaneously as a single unit.

[0139] The development of drug delivery technology of large doses of medicament is currently based on enzymes that facilitate subcutaneous (SC) delivery of co-administered therapeutics. The enzymes work by degrading hyaluronan, which plays a role in resistance to bulk fluid flow in the SC space, limiting rapid, large volume SC drug delivery, dispersion, and absorption. Co-administration of these enzymes (e.g., the proprietary recombinant human hyaluronidase PH20 enzyme (e.g., rHuPH20; Halozyme, Inc.), animal hyaluronidases and variants of hyaluronidases) with therapies can help to overcome administration time and volume barriers associated with existing SC therapeutic formulations, and has been shown to reduce the burden on patients and healthcare providers compared with intravenous formulations. This has countless applications in the current field of injectable therapies by increasing the dispersion and absorption of other injected drugs, such as anticancer therapies (e.g., trastuzumab and rituximab), immunodeficiency treatment, and in subcutaneous urography for improving resorption of radiopaque agents.

[0140] With the development of enzymes which locally degrade hyaluronan in the SC space, the traditional limitations of biological uptake factors that limit injection speed may be significantly reduced. Meaning higher volumes of medicaments can be delivered. Pairing this recombinant human hyaluronidase PH20 enzyme with an angled injection may allow for injection of higher volumes than the current configuration of vertical injection.

[0141] The platform attachment 10 may be coupled to an auto-injector 22 for handheld delivery of the medicament. The auto-injector 22 may be the injection apparatus 18 containing the medicament. The auto-injector 22 may have a needle 21 coupled to a distal end thereof. The autoinjector 22 may include a prefilled syringe filled with the volume of medicament to be delivered.

[0142] The auto-injector 22 may contain 3-5 mL of the medicament. The auto-injector 22 may contain 5-10 mL of the medicament. The auto-injector 22 may contain 10-15 mL of the medicament. The auto-injector 22 may contain 15-20 mL of the medicament. The auto-injector 22 may contain 20- 25 mL of the medicament. The auto-injector 22 may contain 25-30 mL of the medicament. The autoinjector 22 may contain 30-35 mL of the medicament. The auto-injector 22 may contain 35-40 mL of the medicament. The auto-injector 22 may contain 40-45 mL of the medicament. The auto-injector 22 may contain 45-50 mL of the medicament. The auto-injector 22 may contain 50-55 mL of the medicament.

[0143] The auto-injector 22 may contain 5-15 mL of the medicament. The auto-injector 22 may contain 10-20 mL of the medicament. The auto-injector 22 may contain 15-25 mL of the medicament.The auto-injector 22 may contain 20-30 mL of the medicament. The auto-injector 22 may contain 25- 35 mL of the medicament. The auto-injector 22 may contain 30-40 mL of the medicament. The autoinjector 22 may contain 35-45 mL of the medicament. The auto-injector 22 may contain 40-50 mL of the medicament. The auto-injector 22 may contain 45-55 mL of the medicament.

[0144] The auto-injector 22 may contain at least 3 mL. The auto-injector may contain at least 5 mL of the medicament. The auto-injector 22 may contain at least 10 mL of the medicament. The auto-injector 22 may contain at least 15 mL of the medicament. The auto-injector 22 may contain at least 20 mL of the medicament. The auto-injector 22 may contain at least 25 mL of the medicament. The auto-injector 22 may contain at least 30 mL of the medicament. The auto-injector 22 may contain at least 35 mL of the medicament. The auto-injector 22 may contain at least 40 mL of the medicament. The auto-injector 22 may contain at least 45 mL of the medicament. The auto-injector 22 may contain at least 50 mL of the medicament. The auto-injector 22 may contain at least 55 mL of the medicament.

[0145] The housing 16 may extend at an oblique angle relative to the base 12. The housing 16 may extend at an angle A of 0-5 degrees relative to the base 12. The housing 16 may extend at an angle A of 5-10 degrees relative to the base 12. The housing 16 may extend at an angle A of 10-15 degrees relative to the base 12. The housing 16 may extend at an angle A of 15-20 degrees relative to the base 12. The housing 16 may extend at an angle A of 20-25 degrees relative to the base 12, as shown in Figs. 2-4. The housing 16 may extend at an angle A of 25-30 degrees relative to the base 12. The housing 16 may extend at an angle A of 30-35 degrees relative to the base 12. The housing 16 may extend at an angle A of 35-40 degrees relative to the base 12. The housing 16 may extend at an angle A of 40-45 degrees relative to the base 12. The housing 16 may extend at an angle A of 20- 30 degrees relative to the base 12. The housing 16 may extend at an angle A of 30-40 degrees relative to the base 12. The housing 16 may extend at an angle A of 25-35 degrees relative to the base 12. The housing 16 may extend at an angle A of 35-45 degrees relative to the base 12.

[0146] The housing 16 may extend at an angle A of 5 degrees relative to the base 12. The housing 16 may extend at an angle A of 10 degrees relative to the base 12. The housing 16 may extend at an angle A of 15 degrees relative to the base 12. The housing 16 may extend at an angle A of 20 degrees relative to the base 12. The housing 16 may extend at an angle A of 25 degrees relative to the base 12. The housing 16 may extend at an angle A of 30 degrees relative to the base 12. The housing 16 may extend at an angle A of 35 degrees relative to the base 12. The housing 16 may extend at an angle A of 40 degrees relative to the base 12. The housing 16 may extend at an angle A of 45 degrees relative to the base 12.

[0147] The conduit 20 of the housing 16 may include threads configured to engage corresponding threads on the injection apparatus 18. The housing 16 and the injection apparatus 18 may be removably coupled to one another using a threaded coupling. The housing 16 and the injection apparatus 18 may be removably coupled to one another using a snap-fit coupling.

[0148] The injection apparatus 18 may be an injection line 24 or other tubing line or infusion set. The injection line may be coupled to a syringe pump to facilitate transport of the medicament therethrough. Referring to Fig. 4, the injection line 24 may be coupled to the housing 16 by a threaded coupling. The slotted handle 40 may couple to the base 12 of the needle 21 of the injection line 24 to ensure orientation of the housing 16 and the injection line 24.

[0149] The platform attachment 10 may have an injection line 24 for injections requiring constant delivery pressure. The injection line 24 may be the injection apparatus 18 containing the medicament. The injection line 24 may have a prefilled syringe filled with the volume of medicament to be delivered.

[0150] The injection line 24 may contain 3-5 mL of the medicament. The injection line 24 may contain 5-10 mL of the medicament. The injection line 24 may contain 10-15 mL of the medicament. The injection line 24 may contain 15-20 mL of the medicament. The injection line 24 may contain 20-25 mL of the medicament. The injection line 24 may contain 25-30 mL of the medicament. The injection line 24 may contain 30-35 mL of the medicament. The injection line 24 may contain 35-40 mL of the medicament. The injection line 24 may contain 40-45 mL of the medicament. The injection line 24 may contain 45-50 mL of the medicament. The injection line 24 may contain 50-55 mL of the medicament.

[0151] The injection line 24 may contain 5-15 mL of the medicament. The injection line 24 may contain 10-20 mL of the medicament. The injection line 24 may contain 15-25 mL of the medicament. The injection line 24 may contain 20-30 mL of the medicament. The injection line 24 may contain 25-35 mL of the medicament. The injection line 24 may contain 30-40 mL of the medicament. The injection line 24 may contain 35-45 mL of the medicament. The injection line 24 may contain 40-50 mL of the medicament. The injection line 24 may contain 45-55 mL of the medicament.

[0152] The injection line 24 may contain at least 5 mL of the medicament. The injection line 24 may contain at least 10 mL of the medicament. The injection line 24 may contain at least 15 mL of the medicament. The injection line 24 may contain at least 20 mL of the medicament. The injection line 24 may contain at least 25 mL of the medicament. The injection line 24 may contain at least 30 mL of the medicament. The injection line 24 may contain at least 35 mL of the medicament. The injection line 24 may contain at least 40 mL of the medicament. The injection line 24 may contain atleast 45 mL of the medicament. The injection line 24 may contain at least 50 mL of the medicament. The injection line 24 may contain at least 55 mL of the medicament.

[0153] Referring to Fig. 7, there is shown a second embodiment of the platform attachment for angled SC drug delivery, generally designated 10. The platform attachment 10 is similar to the first embodiment of the platform attachment 10 except that the distal surface of the base 12 may include frictional elements to minimize movement of the base 12 when contacting the patient’s skin 13.

[0154] Referring to Fig. 7, the distal surface of the base 12 may comprise a lip 26 extending around a periphery, for increased friction between the base 12 and the skin 13. The lip 26 may protrude distally from the base 12. The lip 26 may be made of a silicone or rubber material. In one embodiment, the lip 26 is made of a plastic material. The base 12 may include a weight proximate a front edge 38 to prevent a slip or other movement during use. The distal surface of the base 12 may comprise one or more ridges 28 protruding distally therefrom, for increased friction between the base 12 and the skin 13. The ridges 28 may be made of a silicone or rubber material. In one embodiment, the ridges 28 are made of a plastic material. The base 12 may include a plurality of protrusions across the base for increased friction between the base 12 and the skin 13. In one embodiment, the proximal surface of the base is a course surface. In one embodiment, the base 12 may include an adhesive to removably affix the platform attachment 10 the patient’s skin 13.

[0155] Referring to Figs. 2-4, there is illustrated a use case of a method of subcutaneously administering a medicament in accordance with an exemplary embodiment of the present disclosure. The method may include attaching to a patient’s skin 13 a platform attachment 10 having a housing 16 with an aperture 14 extending therethrough and a generally flat base 12 angled relative to the housing 16. For example, in Fig. 2 the platform attachment 10 may be attached to a patient’s skin 13. The method may further include attaching to the housing 16 an injection apparatus 18 containing a medicament such that a needle 21 extends distally through the aperture 14 and into the patient’s skin 13. For example, in Fig. 3 the injection apparatus 18 is attached to the housing 16. In one embodiment, attaching the injection apparatus 18 to the housing 16 inserts the needle 21 subcutaneously in the patient’s skin 13 at an angle A of 30 degrees relative to the base 12. The method may further include delivering more than 3 mL of the medicament through the needle 21. In one embodiment, attaching the injection apparatus 18 to the housing 16 comprises attaching an autoinjector 22 to the housing 16. In one embodiment, attaching the injection apparatus 18 to the housing 16 comprises attaching an injection line 24 to the housing 16.

[0156] It will be appreciated by those skilled in the art that changes could be made to the exemplary embodiments shown and described above without departing from the broad inventiveconcepts thereof. It is to be understood that the embodiments and claims disclosed herein are not limited in their application to the details of construction and arrangement of the components set forth in the description and illustrated in the drawings. Rather, the description and the drawings provide examples of the embodiments envisioned. The embodiments and claims disclosed herein are further capable of other embodiments and of being practiced and carried out in various ways.Specific features of the exemplary embodiments may or may not be part of the claimed invention and various features of the disclosed embodiments may be combined. Unless specifically set forth herein, the terms “a”, “an” and “the” are not limited to one element but instead should be read as meaning “at least one”. Finally, unless specifically set forth herein, a disclosed or claimed method should not be limited to the performance of their steps in the order written, and one skilled in the art can readily appreciate that the steps may be performed in any practical order.Definitions

[0100] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs. All patents and publications referred to herein are incorporated by reference in their entireties.

[0101] As used herein, the terms “administer,” “administration,” or “administering” refer to (1) providing, giving, dosing, and / or prescribing by either a health practitioner or his authorized agent or under his or her direction according to the disclosure, and / or (2) putting into, taking, or consuming by a subject, for example a mammal, including a human, according to the disclosure.

[0102] The terms “co-administration,” “co-administering,” “administered in combination with,” “administering in combination with,” “simultaneous,” and “concurrent,” as used herein, encompass administration of two or more active pharmaceutical ingredients to a subject so that both active pharmaceutical ingredients and / or their metabolites are present in the subject at the same time. Coadministration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition in which two or more active pharmaceutical ingredients are present. In some embodiments, simultaneous administration in separate compositions and administration in a composition in which both agents are present are preferred.

[0103] The term “effective amount” or “therapeutically effective amount” refers to that amount of a compound or combination of compounds as described herein that is sufficient to effect the intended application including, but not limited to, disease treatment. A therapeutically effective amount may vary depending upon the intended application (in vitro or in vivo), or the subject and disease condition being treated (e.g., the weight, age and gender of the subject), the severity of the disease condition,the manner of administration, etc., which can readily be determined by one of ordinary skill in the art. The term also applies to a dose that will induce a particular response in target cells (e.g., the reduction of platelet adhesion and / or cell migration). The specific dose will vary depending on the subject to whom the dose is to be administered, the particular compounds chosen, the dosing regimen to be followed, whether the compound is administered in combination with other compounds, timing of administration, the tissue to which it is administered, and the physical delivery system in which the compound is carried.

[0104] A “therapeutic effect” as that term is used herein, encompasses a therapeutic benefit and / or a prophylactic benefit. A prophylactic effect includes delaying or eliminating the appearance of a disease or condition, delaying, or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof.

[0105] As used herein, the terms “treat,” “treatment,” and / or “treating” may refer to the management of a disease, disorder, or pathological condition, or symptom thereof with the intent to cure, ameliorate, stabilize, and / or control the disease, disorder, pathological condition, or symptom thereof. Regarding control of the disease, disorder, or pathological condition more specifically, “control” may include the absence of condition progression, as assessed by the response to the methods recited herein, where such response may be complete (e.g., placing the disease in remission) or partial (e.g., lessening or ameliorating any symptoms associated with the condition). As used herein, the terms “prevent,” “preventing,” and / or “prevention” may refer to reducing the risk of developing a disease, disorder, or pathological condition.

[0106] As used herein, a soluble hyaluronidase is a hyaluronidase of form thereof that is not GPI anchored, and that is soluble under physiological conditions and is secreted upon expression. Soluble hyaluronidases include any that, upon expression, are secreted from a cell and exist in soluble form. Human PH20 hyaluronidase does not occur as a soluble hyaluronidase. It is known in the art that removal of all or a part of the GPI anchor results in soluble forms. Such soluble hyaluronidases include, but are not limited to, bacterial soluble hyaluronidases, non-human soluble hyaluronidases, such as bovine PH20 and ovine PH20, human soluble PH20, and variants thereof. Generally soluble forms of PH20 are produced using protein expression systems that facilitate correct N-glycosylation to ensure the polypeptide retains activity, since glycosylation is important for the catalytic activity and stability of hyaluronidases. Such cells include, for example Chinese Hamster Ovary (CHO) cells (e.g., DG44 CHO cells).

[0107] As used herein the term ‘rHuPH20’ refers to the soluble hyaluronidase composition produced upon expression in a mammalian cell, such as a CHO cell, or other cell that effects glycosylation, ofnucleic acid encoding residues 36-482 of SEQ ID NO: 1. For expression in cells the encoding nucleic acid is linked to the native (residues 1-35 of SEQ ID NO: 1) or a heterologous signal sequence for trafficking and secretion of the encoded polypeptides. The resulting secreted soluble glycoprotein is a heterogeneous mixture of polypeptides, including polypeptides that terminate at residues 479, 480, 481, and 482, and are composed of residues 36-479, 36-480, 36-481, and 36-482 with reference to SEQ ID NO: 1. Shorter C-terminally truncated forms also may be included, in various abundance. Typically, rHuPH20 is produced in cells that facilitate correct N-glycosylation to retain activity, such as CHO cells (e.g., DG44 CHO cells). In some embodiments, one of the most abundant species is the 446 amino acid polypeptide corresponding to residues 36-481 of SEQ ID NO: 1. Also included are polypeptides that are soluble or secreted upon expression in a mammalian cell and have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with residues 36-482 of SEQ ID NO: 1.

[0108] As used herein, “combination therapy” refers to a treatment in which a subject if given two or more therapeutic agents, such as at least two or at least three therapeutic agents, for treating a single disease.

[0109] As used herein, “hyaluronidase activity” refers to the ability to enzymatically catalyze the cleavage of hyaluronic acid. The United States Pharmacopeia (USP) XXII assay for hyaluronidase determines hyaluronidase activity indirectly by measuring the amount of higher molecular weight hyaluronic acid, or hyaluronan, (HA) substrate remaining after the enzyme is allowed to react with the HA for 30 min at 37 °C (USP XXII-NF XVII (1990) 644-645 United States Pharmacopeia Convention, Inc, Rockville, MD). A Reference Standard solution can be used in an assay to ascertain the relative activity, in units, of any hyaluronidase. In vitro assays to determine the hyaluronidase activity of hyaluronidases, such as PH20, including soluble PH20 and esPH20, are known in the art and described herein. Exemplary assays include the microturbidity assay that measures cleavage of hyaluronic acid by hyaluronidase indirectly by detecting the insoluble precipitate formed when the uncleaved hyaluronic acid binds with serum albumin and the biotinylated-hyaluronic acid assay that measures the cleavage of hyaluronic acid indirectly by detecting the remaining biotinylated- hyaluronic acid non-covalently bound to microtiter plate wells with a streptavidin-horseradish peroxidase conjugate and a chromogenic substrate. Reference Standards can be used, for example, to generate a standard curve to determine the activity in Units of the hyaluronidase being tested.

[0110] “Pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” or “physiologically compatible” carrier or carrier medium is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delayingagents, and inert ingredients. The use of such pharmaceutically acceptable carriers or pharmaceutically acceptable excipients for active pharmaceutical ingredients is well known in the art. Except insofar as any conventional pharmaceutically acceptable carrier or pharmaceutically acceptable excipient is incompatible with the active pharmaceutical ingredient, its use in the therapeutic compositions of the invention is contemplated. Additional active pharmaceutical ingredients, such as other drugs, can also be incorporated into the described compositions and methods.

[0111] As used herein, “specific activity” refers to Units of activity per mg protein. The milligrams of hyaluronidase is defined by the absorption of a solution of at 280 nm assuming a molar extinction coefficient of approximately 1.7, in units of M'1cm'1.

[0112] As used herein, “neutral active” refers to the ability of a PH20 polypeptide to enzymatically catalyse the cleavage of hyaluronic acid at neutral pH (e.g., at or about pH 7.0).

[0113] As used herein, a “GPI-anchor attachment signal sequence” is a C-terminal sequence of amino acids that directs addition of a preformed GPI-anchor to the polypeptide within the lumen of the ER. GPI-anchor attachment signal sequences are present in the precursor polypeptides of GPI-anchored polypeptides, such as GPI-anchored PH20 polypeptides. The C-terminal GPI-anchor attachment signal sequence typically contains a predominantly hydrophobic region of 8-20 amino acids, preceded by a hydrophilic spacer region of 8-12 amino acids, immediately downstream of the co-site, or site of GPI-anchor attachment. GPI-anchor attachment signal sequences can be identified using methods well known in the art, such as but not limited to, in silico methods and algorithms (see, e.g., Udenfriend et al. (1995) Methods Enzymol. 250:571-582, Eisenhaber et al., (1999) J. Biol. Chem. 292: 741-758, Fankhauser et al., (2005) Bioinformatics 21 :1846-1852, Omaetxebarria et al., (2007) Proteomics 7: 1951-1960, Pierleoni et al., (2008) BMC Bioinformatics 9:392), including those that are readily available on bioinformatic websites, such as the ExPASy Proteomics tools site (e.g., the World Wide Web site expasy.ch / tools / ).

[0114] As used herein, “sequence identity” refers to the relatedness between or among polypeptides among nucleic acid molecules. Sequence identity can be assessed by aligning two sequences and counting the number of differences between the aligned portion and the sequence to which it is compared. Whether any two molecules have nucleotide sequences or amino acid sequences that are at least 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% “identical” or “homologous” can be determined using known computer algorithms such as the “FASTA” program, using for example, the default parameters as in Pearson et al. (1988) Proc. Natl. Acad. Sci. USA 85:2444 (other programs include the GCG program package (Devereux, J., et al., Nucleic Acids Research 12(I):387 (1984)),BLASTP, BLASTN, FASTA (Altschul, S.F., et al., J Mol Biol 215:403 (1990)); Guide to Huge Computers, Martin J. Bishop, ed., Academic Press, San Diego, 1994, and Carrillo et al. (1988) SIAM J Applied Math 48: 1073). For example, the BLAST function of the National Center for Biotechnology Information database can be used to determine identity. Other commercially or publicly available programs include, DNAStar “MegAlign” program (Madison, WI) and the University of Wisconsin Genetics Computer Group (UWG) “Gap” program (Madison WI). Percent homology or identity of proteins and / or nucleic acid molecules can be determined, for example, by comparing sequence information using a GAP computer program (e.g., Needleman et al. (1970) J. Mol. Biol. 48:443, as revised by Smith and Waterman ((1981) Adv. Appl. Math. 2:482). Briefly, the GAP program defines similarity as the number of aligned symbols (i.e., nucleotides or amino acids), which are similar, divided by the total number of symbols in the shorter of the two sequences. Default parameters for the GAP program can include: (1) a unary comparison matrix (containing a value of 1 for identities and 0 for non identities) and the weighted comparison matrix of Gribskov et al. (1986) Nucl. Acids Res. 14:6745, as described by Schwartz and Dayhoff, eds., ATLAS OF PROTEIN SEQUENCE AND STRUCTURE, National Biomedical Research Foundation, pp. 353 358 (1979); (2) a penalty of 3.0 for each gap and an additional 0.10 penalty for each symbol in each gap; and (3) no penalty for end gaps.

[0115] Therefore, as used herein, the term “identity” or “homology” represents a comparison between a test and a reference polypeptide or polynucleotide.

[0116] As used herein, the term at least “90% identical to” refers to percent identities from 90 to 99.99 relative to the reference nucleic acid or amino acid sequence of the polypeptide. Identity at a level of 90% or more is indicative of the fact that, assuming for exemplification purposes a test and reference polypeptide length of 100 amino acids are compared. No more than 10% (i.e., 10 out of 100) of the amino acids in the test polypeptide differs from that of the reference polypeptide. Similar comparisons can be made between test and reference polynucleotides. Such differences can be represented as point mutations randomly distributed over the entire length of a polypeptide or they can be clustered in one or more locations of varying length up to the maximum allowable, e.g., 10 / 100 amino acid difference (approximately 90% identity). Differences are defined as nucleic acid or amino acid substitutions, insertions or deletions. At the level of homologies or identities above about 85-90%, the result should be independent of the program and gap parameters set; such high levels of identity can be assessed readily, often by manual alignment without relying on software.

[0117] As used herein, an “aligned sequence” refers to the use of homology (similarity and / or identity) to align corresponding positions in a sequence of nucleotides or amino acids. Typically, two or moresequences that are related by 50% or more identity are aligned. An aligned set of sequences refers to 2 or more sequences that are aligned at corresponding positions and can include aligning sequences derived from RNAs, such as ESTs and other cDNAs, aligned with genomic DNA sequence.

[0118] As used herein, “denaturing condition” or “denaturation condition” refers to any condition or agent that, when exposed to a protein, affects or influences the degradation or denaturation of the protein, generally as a result of a loss or partial loss of the tertiary or secondary structure of the protein. Denaturing conditions can result in effects such as loss or reduction in activity, loss or reduction of solubility, aggregation and / or crystallization.

[0119] As used herein, “resistance to a denaturation condition” refers to any amount of decreased reduction or elimination of a property or activity of the protein associated with or caused by denaturation. For example, denaturation is associated with or causes increased crystallization or aggregation, reduced solubility or decreased activity. Hence, resistance to denaturation means that the protein exhibits decreased aggregation or crystallization, increased solubility or increased or greater activity (e.g., hyaluronidase activity) when exposed to a denaturing condition compared to a reference protein (e g., unmodified enzyme).

[0120] As used herein, “stability of a modified PH20 hyaluronidase” means that it exhibits resistance to denaturation caused by a denaturation condition or denaturing agent.

[0121] When ranges are used herein to describe, for example, physical or chemical properties such as molecular weight or chemical formulae, all combinations and subcombinations of ranges and specific embodiments therein are intended to be included. Use of the term “about” when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range may vary. The variation is typically from 0% to 15%, from 0% to 10%, from 0% to 5%, or the like, of the stated number or numerical range.

[0122] As used herein, the term “about” means that amounts, sizes, formulations, parameters, shapes and other quantities and characteristics are not, and need not be exact, but may be approximate and / or larger or smaller, as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the art. In general, an amount, size, formulation, parameter, shape or other quantity or characteristic is “about” or “approximate” whether or not expressly stated to be such. The term “about” generally refers to a particular numeric value that is within an acceptable error range as determined by one of ordinary skill in the art, which will depend in part on how the numeric value is measured or determined, i.e., the limitations of themeasurement system. For example, “about” can mean a range of ±20 %, ±10 %, or ±5 % of a given numeric value.

[0123] The transitional terms “comprising,” “consisting essentially of,” and “consisting of,” when used in the appended claims, in original and amended form, define the claim scope with respect to what unrecited additional claim elements or steps, if any, are excluded from the scope of the claim(s). The term “comprising” is intended to be inclusive or open-ended and does not exclude any additional, unrecited element, method, step, or material. The term “consisting of’ excludes any element, step or material other than those specified in the claim and, in the latter instance, impurities ordinarily associated with the specified material(s). The term “consisting essentially of’ limits the scope of a claim to the specified elements, steps, or material(s) and those that do not materially affect the basic and novel character! stic(s) of the claimed invention. All compounds, compositions, formulations, and methods described herein that embody the present invention can, in alternate embodiments, be more specifically defined by any of the transitional terms “comprising,” “consisting essentially of,” and “consisting of.” The term “comprising” (and related terms such as “comprise” or “comprises” or “having” or “including”) includes those embodiments such as, for example, an embodiment of any composition of matter, method, or process that “consist of’ or “consist essentially of’ the described features.

[0124] The present disclosure generally relates to automatic or manual triggering devices that are used in the delivery of high volumes of injectable fluid, such as medicament, (e.g., 2 ml, 3 ml, 5 ml, 10 ml or higher doses) for sub skin surface penetration (e.g., subcutaneous and intramuscular injections). Devices that deliver high volumes of medicament are prone to configuration restrictions as the delivery volume of the medicament increases, such as spring force limitations and syringe container breakage. Due to the current configurations of overall device design, and the limitations of the biological uptake factors that limit injection speed and volume, injection devices would be held for long durations, and often result in other delivery methods being used such as on-body delivery systems which are attached to the patient during delivery.

[0125] Increasingly, more biologies are allowing for at-home administration for patients. However, for many biologies, high doses and the resulting high volume of medicament that must be delivered often precludes self-administration because of the length of time required to hold the delivery device in place. Protein hyper-concentration can be used to reduce injection volume, but the resulting medicament often has a much higher viscosity than the traditional biologies. High powered injectors may allow delivery of these hyper-concentrated proteins. However, typical handheld injector designs only allow for up to 2.25 mL of a viscous medicament to be injected in 30 seconds.

[0126] With the development of enzymes which locally degrade hyaluronan (HA) in the subcutaneous (SC) space, thereby temporarily removing a barrier to fluid flow, the traditional limitations of biological uptake factors that limit injection speed and volume may be reduced. As such, higher volumes of viscous medicaments may be delivered. Accordingly, there is a need to provide a handheld device capable of delivering a high dose (e.g., 3 mL, 5 mL, 10 mL, 20 mL and up to 50 mL) of a viscous medicament in a delivery time appropriate for a handheld device.

[0127] Drug delivery technology of high doses of a viscous medicament is currently based on the proprietary recombinant human hyaluronidase PH20 enzyme (e.g., rHuPH20; Halozyme, Inc.) that facilitates SC delivery of co-administered therapeutics. rHuPH20 works by degrading the HA, thus decreasing the resistance to bulk fluid flow in the SC space, and permitting high volume SC drug delivery, dispersion, and absorption. Co-admini strati on of rHuPH20 with injectable therapies can overcome administration time and volume barriers associated with existing SC therapeutic formulations and has been shown to reduce the burden on patients and healthcare providers compared with intravenous formulations. rHuPH20 has countless applications in the current field of injectable therapies by increasing the dispersion and absorption of other injected drugs, such as anticancer therapies (e.g., trastuzumab and rituximab), immunodeficiency treatment, in subcutaneous urography for improving resorption of radiopaque agents, and fluid delivery for rehydration.Formulation for High Dose Injection

[0128] The present disclosure provides a formulation for a high dose injection comprising a hyaluronidase enzyme. In one embodiment, the formulation is an aqueous formulation. In one embodiment, the formulation comprises one or more pharmaceutically acceptable carriers.Forms of Soluble Hyaluronidases

[0129] Soluble hyaluronidases include any that, upon expression, are secreted from a cell and exist in soluble form. Such soluble hyaluronidases include, for example, but are not limited to, bacterial soluble hyaluronidases, non-human soluble hyaluronidases, such as bovine PH20 and ovine PH20, human soluble PH20, and variants thereof. Generally soluble forms of PH20 are produced using protein expression systems that facilitate correct N-glycosylation to ensure the polypeptide retains activity, since glycosylation is important for the catalytic activity and stability of hyaluronidases. Such cells include, for example Chinese Hamster Ovary (CHO) cells (e.g., DG44 CHO cells).

[0130] Vorhyaluronidase alfa is a recombinant human hyaluronidase PH-20 analog corresponding to the amino acid sequence of human hyaluronidase PH-20 at positions 36-482. Vorhyaluronidase alfa is produced in Chinese hamster ovary cells. Vorhyaluronidase alfa is a glycoprotein (molecular weight: 60,000 - 65,000) consisting of 447 amino acid residues. In one embodiment, the soluble PH20product is the composition produced by expression of nucleic acid encoding residues 36-482 in CHO cells resulting in a mixture of polypeptides with C-termini at residues 477, 478, 479, 480, 481, and 482.

[0131] Soluble PH20 hyaluronidase is available and sold, for example, under the trademark ENHANZE® (CAS name of 36-482-Hyaluronoglucosaminidase PH20 (human)). ENHANZE® is the mixture of polypeptides produced by expression of nucleic acid encoding amino acids 36-482 (SEQ ID NO: 2). The product is a mixture of polypeptides, produced by expression of nucleic acid encoding amino acids 36-477 (SEQ ID NO: 3), 36-478 (SEQ ID NO: 4), 36-479 (SEQ ID NO: 5), 36-480 (SEQ ID NO: 6), 36-481 (SEQ ID NO: 7), and 36-482 (SEQ ID NO: 2). ENHANZE® technology provides to a drug delivery technology, employing the soluble hyaluronidases to facilitate the delivery of injected drugs and fluids. When co-formulated with other drugs or administered with other drugs, the ENHANZE® technology reduces treatment burden for patients. It can allow for large volume subcutaneous injection with increased dispersion and absorption of co-administered therapies.

[0132] In one embodiment, one or more of N47, N131, N200, N219, N333, N358, or T440 in SEQ ID NOs: 2-7 are glycosylation sites. In one embodiment, one or more of Q444, 1445, F446, or Y447 in SEQ ID NOs: 2-7 are partial processing sites. In one embodiment, one or more of C25-C316, C189-C203, C341-C352, C346-C400, C402-C408, or C423-C429 in SEQ ID NOs.: 2-7 are disulfides.

[0133] rHuPH20 refers to the composition produced upon expression in a cell, such as CHO cell, of nucleic acid encoding residues 36-482 of SEQ ID NO: 8, generally linked to the native or a heterologous signal sequence (residues 1-35 of SEQ ID NO: 8). rHuPH20 is produced by expression of a nucleic acid molecule, such as encoding amino acids 1-482 (set forth in SEQ ID NO: 8) in a mammalian cell. Translational processing removes the 35 amino acid signal sequence. As produced in the culture medium there is heterogeneity at the C-terminus such that the product, designated rHuPH20, includes a mixture of species that can include any one or more of the polypeptides 36-480, 36-481, and 36-482 of SEQ ID NO: 8, and some shorter polypeptides, in various abundance. rHuPH20 and forms of soluble hyaluronidase are produced in cells, such as CHO cells, for example DG44 CHO cells, that facilitate N-glycosylation. PH20 is a glycoprotein, and as known in the art, requires glycosylation retain activity. See, e.g, U.S. Patent Nos. 8,927,249 and 9,284,543 (and PCT Publication No. WO 2010 / 077297), which describe the effects of glycosylation and partial glycosylation and elimination of glycosylation on the activity of soluble forms of PH20. These patents and publications also describe and exemplify soluble C-terminally truncated forms of PH20.Forms of Soluble Human PH20

[0134] Soluble hyaluronidases include bovine and ovine PH20, and recombinant and humanized forms thereof. Human PH20 in nature includes a GPI anchor and exists linked to sperm cells; it is not soluble. C-terminally-truncated forms thereof are soluble. Soluble forms of recombinant human PH20 have been produced and can be used in the compositions, combinations and methods described herein. Descriptions of and production of such soluble forms of PH20 are described, for example, in U.S. PatentNos. 7,767,429, 8,202,517, 8,431,380, 8,431,124, 8,450,470, 8,765,685, 8,772,246, 7,871,607, 7,846,431, 7,829,081, 8,105,586, 8,187,855, 8,257,699, 8,580,252, 9,677,061, and 9,677,062, each incorporated by reference herein. The soluble hyaluronidases, thus include forms of human PH20, which ae neutral active hyaluronidases and which require glycosylation for activity.

[0135] SEQ ID NO: 1 sets forth the sequence of the precursor polypeptides; the mature PH20 polypeptide (residues 36-509); soluble forms also include those with amino acid truncations at the N- terminal, such as deletions of the first one, two, three, or fours residues, such that the resulting polypeptides have an N-terminus, for example, at residue 36, 37, 38, 39, or 40, and a C-terminus at a residue from 465 to 500, and variants thereof, including, but not limited to, variants discussed below, variants known in the art, and allelic variants.

[0136] Hyaluronidases for use in the compositions, combinations and methods herein are soluble neutral active hyaluronidases. Exemplary thereof are the soluble C-terminally truncated forms of mature human PH20. Soluble forms that have hyaluronidase activity, include but are not limited to, those that are truncated at residues from 465 to 500 of SEQ ID NO: 1, and that are, upon expression, secreted. Exemplary thereof are polypeptides that have sequence 36-465, 36-466, 36-467, 36-468, 36-469, 35-470, 36-471, 36-472, 36-474, 36-475, 36-476, 35-477, 36-478, 36-479, 36-480, 36-481, 36-482, 36-483 35-484, 36-485, 36-486, 36-487, 36-488, 36-489, 36-490, 35-491, 36-492, 36-493, 36-494, 36-495, 36-496, 36-497, 35-498, 36-499, and 36-500 of SEQ ID NO: 1, as well as N- terminally truncated forms of each of the preceding that lack two to five residues at the N-terminus, such as for example 37-368, 38-468, and any others that exhibit hyaluronidase activity at neutral pH, such as pH in the range of 7.0-7.4.

[0137] Thus, such soluble forms include truncated forms of the mature form of human PH20 lacking all or a portion of the C-terminal GPI anchor, so long as the hyaluronidase is soluble and retains hyaluronidase activity. Soluble forms are secreted upon expression in mammalian cells, and are encoded with a signal sequence, such are residues 1-35 of SEQ ID NO. 1 or a heterologous signal sequence that is cleaved by the cell to effect secretion. Soluble forms are forms that, when expressed in a cell, lack the signal peptide. Also included among soluble hyaluronidases are variants of the soluble PH20 polypeptides that exhibit hyaluronidase activity. Variants include polypeptides havingat least 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to any of the PH20 polypeptides 36-465, 36-466, 36-467, 36-468, 36-469, 35-470, 36-471, 36-472, 36-474, 36-475, 36-476, 35-477, 36-478, 36-479, 36-480, 36-481, 36-482, 36-48335-484, 36-485, 36-486, 36-487, 36-488, 36-489, 36-490, 35-491, 36-492, 36-493, 36-494, 36-495,36-496, 36-497, 35-498, 36-499, and 36-500 of SEQ ID NO: 1. Amino acid variants include conservative and non-conservative insertions, or deletions, or replacements, and include the modifications, singly or combinations of the modifications detailed, for example, in U.S. Patent No. 11,041,149 and International PCT publication No. WO 2013 / 102144. U.S. Patent No. 11,041,149 and International PCT publication No. WO 2013 / 102144 describe a systematic analysis and results identifying the effects of amino acid modifications at each residue in PH20 to thereby provide a structure / function map of PH20; a skilled person can identify replacement residues and consequent alterations in properties and activities, such as for effecting increases in enzymatic activity, stability in denaturing conditions, and also residues whose replacement or deletion decreases or eliminates enzymatic activity.

[0138] It is understood that residues that are important or otherwise required for the activity of a hyaluronidase, such as any described above or known to skill in the art, are generally invariant and, except for possible conservative amino acid substitutions, cannot be changed. These include, for example, active site residues. For example, amino acid residues 111, 113 and 176 (corresponding to residues in the mature PH20 polypeptide) of a human PH20 polypeptide, or soluble form thereof, are generally invariant and are not altered. Other residues that confer glycosylation and formation of disulfide bonds required for proper folding also can be invariant.

[0139] The soluble human PH20 hyaluronidase is GPI-anchored and is rendered soluble by truncation at the C-terminus by removal of all or a part of the GPI anchor. Such truncation can remove all of the GPI anchor attachment sequence or can remove only some of the GPI anchor attachment sequence. The resulting polypeptide, however, is soluble. In instances where the soluble hyaluronidase retains a portion of the GPI anchor attachment signal sequence, 1, 2, 3, 4, 5, 6, 7 or more amino acid residues in the GPI anchor attachment signal sequence can be retained, provided the polypeptide is soluble. Polypeptides containing one or more amino acids of the GPI anchor are termed extended soluble hyaluronidases. One of skill in the art can determine whether a polypeptide is GPI-anchored using methods well known in the art. Such methods include, but are not limited to, using known algorithms to predict the presence and location of the GPI anchor attachment signal sequence and co-site, and performing solubility analyses before and after digestion with phosphatidylinositol-specific phospholipase C (PI-PLC) or D (PI-PLD).

[0140] Extended soluble hyaluronidases, which terminate for example, at residues 495, 496, 497, 498, 499, and 500, with reference to SEQ ID NO: 1, such as those set forth in SEQ ID NO: 9 (residues 1- 495), SEQ ID NO: 10 (residues 1-496), SEQ ID NO: 1 1 (residues 1-497), SEQ ID NO: 12 (residues 1-498), SEQ ID NO: 13 (residues 1-499), and SEQ ID NO: 14 (residues 1-500), can be produced by making C-terminal truncations to any naturally GPI-anchored hyaluronidase such that the resulting polypeptide is soluble and contains one or more amino acid residues from the GPI anchor attachment signal sequence (see, e.g., U.S. Patent No. 8,927,249). These include hyaluronidases that are neutral active, soluble, contain amino acid substitutions, and have at least 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% or more sequence identity to any of SEQ ID NOs: 9-14.

[0141] Typically, for use in the compositions, combinations, and methods herein, a soluble human hyaluronidase, such as a soluble human PH20, is used, such as a PH20 and variants having, for example, at least 91% or 95% or 98% sequence identity thereto, including those with 1 to 5 N-terminal residues deleted. Hyaluronidases used in the regimens, combinations, compositions, and methods herein can be recombinantly produced or can be purified or partially purified from natural sources, such as, for example, from testes extracts. Methods for production of recombinant proteins, including recombinant hyaluronidases, are well known in the art.

[0142] Recombinant soluble forms of human PH20 have been generated and can be used in the compositions, combinations and methods provided herein. For example, with reference to SEQ ID NO: 1, which sets forth the sequence of full length precursor PH20, which includes a signal sequence (residues 1-35), soluble forms include, but are not limited to, C-terminal truncated polypeptides of human PH20 set forth in SEQ ID NO: 1 having a C-terminal amino acid residue 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499 or 500 of the sequence of amino acids set forth in SEQ ID NO: 1, or polypeptides that exhibit at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity thereto, when aligned with the unmodified sequence of the soluble PH20, have activity at neutral pH, and are soluble (secreted into the medium when expressed in a mammalian cell). Soluble forms of human PH20 generally include those that contain amino acids 36-464 set forth in SEQ ID NO: 1 and terminate at any of residues, 465-500, and optionally include a 1-3 amino acid deletion at the N-terminus (i.e. lack residues 36, 36-37, or 36-38 of SEQ ID NO: 1). For example, when expressed in mammalian cells, the 35 amino acid N-terminal signal sequence (residues 1-35 of SEQ ID NO: 1) is cleaved during processing, and a soluble form of the protein is secreted. Thus, the mature soluble polypeptides include those that contain amino acids 36 to 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, and up to and including500 of SEQ ID NO: 1. Exemplary of soluble hyaluronidases are soluble human PH20 polypeptides that are 442, 443, 444, 445, 446 or 447 amino acids in length, such as set forth those set forth above, and variants thereof that have, for example, at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity thereto and retains hyaluronidase activity. The generation of such soluble forms of recombinant human PH20 are described, for example, in U.S. Patent Nos. 7,767,429, 8,202,517, 8,431,380, 8,431,124, 8,450,470 8,765,685, 8,772,246, 7,871,607, 7,846,431, 7,829,081, 8,105,586, 8,187,855, 8,257,699, 8,580,252, 9,677,061, and 9,677,062.

[0143] Generally soluble forms of PH20 are produced using protein expression systems that facilitate correct N-glycosylation to ensure the polypeptide retains activity, since glycosylation is important for the catalytic activity and stability of hyaluronidases. Such cells include, for example Chinese Hamster Ovary (CHO) cells (e g. DG44 CHO cells).

[0144] The composition that recombinantly produced from mammalian cells, such as CHO cells, has been referred to rHuPH20. It refers to the composition produced upon expression in a cell, such as CHO cell, of nucleic acid encoding residues 36-482 of SEQ ID NO: 1, generally linked to the native (residues 1-35 of SEQ ID NO: 1) or a heterologous signal sequence. rHuPH20 is produced by expression of a nucleic acid molecule, such as encoding amino acids 1-482 (set forth in SEQ ID NO: 1) or 36 to 482 with a heterologous signal sequence. Post translational processing removes the 35 amino acid signal sequence, resulting in polypeptide or a mixture of polypeptides, including those set forth in SEQ ID NO: 2 (residues 36-482), SEQ ID NO: 3 (residues 36-477), SEQ ID NO: 4 (residues 36-478), SEQ ID NO: 5 (residues 36-479), SEQ ID NO: 6 (residues 36-480), and SEQ ID NO: 7 (residues 36-481). As produced in the culture medium there is heterogeneity at the C-terminus such that the product, designated rHuPH20, includes a mixture of species that can include any one or more of SEQ ID NOs: 3 and 44-49 in various abundance. Generally, the soluble hyaluronidases, rHuPH20 is produced in cells that facilitate correct N-glycosylation to retain activity, such as CHO cells (e.g. DG44 CHO cells). Human soluble PH20 hyaluronidase requires glycosylation for activity. When produced recombinantly from a vector encoding residues 36-582, the most abundant species is the 446 amino acid polypeptides corresponding to residues 36-481 of SEQ ID NO: 1. The particular distribution of resulting polypeptides can depend upon the particular method of production. An exemplary method for production of high levels of PH20 is detailed, for example in U.S. Patent Nos. 8,187,855 and 8,343,487.Glycosylation of Hyaluronidases

[0145] Glycosylation, including N- and O-linked glycosylation, of some hyaluronidases, including the soluble PH20 hyaluronidases, can be important for their catalytic activity and stability. For somehyaluronidases, removal of N-linked glycosylation can result in near complete inactivation of the hyaluronidase activity. For such hyaluronidases, the presence of N-linked glycans can be important for generating an active enzyme.

[0146] N-linked oligosaccharides fall into several primary types (oligomannose, complex, hybrid, sulfated), all of which have (Man) 3-GlcNAc-GlcNAc- cores attached via the amide nitrogen of Asn residues that fall within -Asn-Xaa-Thr / Ser-sequences (where Xaa is not Pro). Glycosylation at an - Asn-Xaa-Cys-site has been reported for coagulation protein C. In some instances, a hyaluronidase, such as a PH20 hyaluronidase, can contain N-glycosidic and O-glycosidic linkages. For example, PH20 has O-linked oligosaccharides as well as N-linked oligosaccharides. There are six potential N- linked glycosylation sites at N82, N166, N235, N254, N368, N393 of human PH20 exemplified in SEQ ID NO: 1.Variants of PH20

[0147] As discussed above, variants of PH20 are known to those of skill in the art, or readily can be prepared in view of the skill and knowledge in the art. Variants include those with amino acid replacements, insertions, and deletions. Variants of the soluble PH20 polypeptides that have altered properties, such as increased stability and / or activity, have been produced. U.S. Patent No. 9,447,401 and family members U.S. Patent Nos. 10,865,400, 11,041,149 and 11,066,656 describe and provide a structure / function map of human PH20 detailing the effects of amino acid replacements at every residue in the catalytic domain of PH20. These patents provide about 7000 examples in which the effects of replacing each amino acid with 15 other amino acids on activity and stability were identified and described. By virtue of those patents, and earlier publications / patents, describing virtually all variants of soluble PH20 polypeptides are known in the art. A skilled person readily can prepare soluble hyaluronidases and variants thereof and know the properties of the resulting hyaluronidase.

[0148] Other variants also are known to those of skill in the art, and can be used in the combinations, regimens, and methods described herein. For example, see, International PCT Publication Nos. W02020 / 022791 and W02020 / 197230, which are incorporated by reference, and which describe modified PH20 polypeptides. These polypeptides, which include variants of the PH20 polypeptides that generally span residues 38-468, and include replacements, insertions, and deletions. The variants include for example one or more amino acid residues changes S343E, I344N, M345T, M348K, K349E, L353A, L354I, N356E, and I361T (with reference to SEQ ID NO: 1), and others, including about 15 amino acid variations, and truncations at the N-terminus and C-terminus. Variants that contain such modifications and others are set forth in SEQ ID NOs: 60-115 of International PCT publication No. W02020 / 022791. Exemplary of these polypeptides is the polypeptide of SEQ IDNO: 99, therein, and reproduced herein as SEQ ID NO: 15. International PCT Publication No. W02021 / 150079 provides variant PH20 polypeptides described as having increased stability relative to unmodified PH20, such as those in rHuPH20. These variant polypeptides have been shown to have PH20 activity and are described as having use for subcutaneous co-administration with other agents.

[0149] In one embodiment, the variant of PH20 is a variant of human PH20 or rHuPH20 selected from any one of SEQ ID NOs: 16-47.

[0150] In one embodiment, the hyaluronidase enzyme is a human hyaluronidase enzyme. Exemplary human hyaluronidase enzymes include HYAL1, HYAL2, HYAL3, HYAL4, HYAL5 (also known as SPAM1 or PH20), and HYAL6 (also known as HYALP1). In one embodiment, the hyaluronidase enzyme is a recombinant hyaluronidase enzyme. In one embodiment, the hyaluronidase enzyme is a recombinant human hyaluronidase enzyme. In one embodiment, the hyaluronidase enzyme is a recombinant human hyaluronidase PH20 enzyme. Exemplary hyaluronidase enzymes that can be used in the disclosure can be found in the following patents and patent applications which are incorporated by reference herein in their entirety: U.S. 2022 / 0289864 (Alteogen), U.S. 9,084,743 (Baxter), U.S. 9,993,529 (Halozyme), U.S. 8,795,654, U.S. 2009 / 0311237 (Greg Frost), U.S. 9,284,543 (Halozyme), U.S. 10,265,410 (Halozyme), U.S. 10,137, 104 (Halozyme), U.S. 2013 / 0022592, U.S. 2019 / 0284263 (Greg Frost), U.S. 11,0653,09, WO 2017 / 185383, U.S. 11,041,149 (Halozyme), U.S. 2010 / 0003238 (Greg Frost), U.S. 8,343,487 (Halozyme), U.S. 10,301,376 (Baxalta), U.S. 2013 / 0344048, U.S. 2021 / 0363270 (Alteogen), U.S. 2021 / 0155913 (Alteogen), WO 2021 / 150079 (Dassault Systems SolidWorks), and WO 2022 / 031093 (Toshiba TEC Kabushiki Kaisha).

[0151] In one embodiment, the hyaluronidase enzyme has an activity of between about 150 U / mL to about 1,000 kU / mL, about 150 U / mL to about 900 kU / mL, about 150 U / mL to about 800 kU / mL, about 150 U / mL to about 700 kU / mL, about 150 U / mL to about 600 kU / mL, about 150 U / mL to about 500 kU / mL, about 150 U / mL to about 400 kU / mL, about 150 U / mL to about 300 kU / mL, about 150 U / mL to about 200 kU / mL, about 500 U / mL to about 200 kU / mL, about 1 kU / mL to about 200 kU / mL, about 10 kU / mL to about 200 kU / mL, about 25 kU / mL to about 200 kU / mL, about 50 kU / mL to about 200 kU / mL, about 100 kU / mL to about 200 kU / mL, about 100 kU / mL to about 150 kU / mL, or about 120 kU / mL. In one embodiment, the hyaluronidase enzyme has an activity of about 10 kU / mL for a 5 mL formulation or about 5 kU / mL for a 10 mL formulation. In one embodiment, the hyaluronidase enzyme has a minimum activity of about 150 U / mL and a maximum activity of about 110,000 U / mL (110 kU / mL). In one embodiment, the hyaluronidase enzyme is recombinant human hyaluronidase PH20 enzyme with an activity of about 120 kU / mL. In another embodiment, thehyaluronidase enzyme is recombinant human hyaluronidase PH20 enzyme with a minimum activity of about 150 U / mL and a maximum activity of about 110,000 U / mL.

[0152] In one embodiment, the concentration of hyaluronidase enzyme in the formulation is between about 10 U / mL to about 50,000 U / mL, about 10 U / mL to about 45,000 U / mL, about 10 U / mL to about 40,000 U / mL, about 10 U / mL to about 35,000 U / mL, about 10 U / mL to about 30,000 U / mL, about 10 U / mL to about 25,000 U / mL, about 10 U / mL to about 20,000 U / mL, about 10 U / mL to about 15,000 U / mL, about 10 U / mL to about 10,000 U / mL, about 100 U / mL to about 9,000 U / mL, about 100 U / mL to about 8,000 U / mL, about 100 U / mL to about 7,000 U / mL, about 100 U / mL to about 6,000 U / mL, about 100 U / mL to about 5,000 U / mL, about 500 U / mL to about 5,000 U / mL, about 500 U / mL to about 4,000 U / mL, about 500 U / mL to about 3,000 U / mL, about 1,000 U / mL to about 3,000 U / mL, about 1,500 U / mL to about 3,000 U / mL, about 1,500 U / mL to about 2,500 U / mL, or about 2,000 U / mL. In one embodiment, the formulation comprises about 1,500 U / mL to about 10,000 U / mL of hyaluronidase enzyme. In one embodiment, the formulation comprises about 1,500 U / mL to about 10,000 U / mL of recombinant human hyaluronidase PH20 enzyme. In one embodiment, the formulation comprises about 2,000 U / mL of recombinant human hyaluronidase PH20 enzyme. In one embodiment, the formulation comprises about 5,000 U / mL of recombinant human hyaluronidase PH20 enzyme. In another embodiment, the formulation comprises at least 4,000 U / mL of recombinant human hyaluronidase PH20 enzyme. In another embodiment, the formulation comprises at least 7,500 U / mL of recombinant human hyaluronidase PH20 enzyme. In another embodiment, the formulation comprises at least 10,000 U / mL of recombinant human hyaluronidase PH20 enzyme. In one embodiment, the formulation comprises about 4,000 U / mL of recombinant human hyaluronidase PH20 enzyme.

[0153] In one embodiment, the formulation comprises a hyaluronidase enzyme that permits a high volume of the formulation to be injected into a subject in need thereof at a high flow rate. In one embodiment, the formulation comprises a hyaluronidase enzyme at a concentration and / or activity that permits a high volume of the formulation to be injected into a subject in need thereof at a high flow rate.

[0154] In one embodiment, the formulation comprises one or more pharmaceutically acceptable additives including, but not limited to, carriers, excipients, fillers, preservatives, stabilizers, and antioxidants. The pharmaceutically acceptable additive can be any pharmaceutically acceptable additive known to a person of skill in the art for an injectable formulation. In one embodiment, the formulation comprises histidine. In one embodiment, the formulation comprises sodium chloride. In one embodiment, the formulation comprises polysorbate. In one embodiment, the polysorbatecomprises polysorbate 80. In one embodiment, the formulation comprises between about 0.001% to about 5%, about 0.001% to about 4.5%, about 0.001% to about 4.0%, about 0.001% to about 3.5%, about 0.001% to about 3.0%, about 0.001% to about 2.5%, about 0.001% to about 2.0%, about 0.001% to about 1.5%, about 0.001% to about 1.0%, about 0.001% to about 0.5%, about 0.005% to about 0.5%, about 0.005% to about 0.1%, about 0.005% to about 0.05%, about 0.01% to about 0.05%, or about 0.02% polysorbate 80. In one embodiment, the formulation comprises an antioxidant. In one embodiment, the antioxidant comprises methionine. In one embodiment, the formulation comprises between about 0.5 mM to about 50 mM, about 0.5 mM to about 45 mM, about 0.5 mM to about 40 mM, about 0.5 mM to about 35 mM, about 0.5 mM to about 30 mM, about 0.5 mM to about 25 mM, about 0.5 mM to about 20 mM, about 5 mM to about 20 mM, about 5 mM to about 15 mM, or about 10 mM antioxidant. In one embodiment, the formulation comprises about 10 mM methionine. In one embodiment, the formulation comprises a carrier protein.

[0155] In one embodiment, the formulation has a pH of between about 4.0 to about 8.0, about 4.2 to about 8.0, about 4.4 to about 7.8, about 4.6 to about 7.8, about 4.6 to about 7.6, about 4.8 to about 7.8, about 5.0 to about 7.8, about 5.2 to about 7.8, about 5.4 to about 7.6, about 5.6 to about 7.4, about 5.8 to about 7.2, about 6.0 to about 7.0, about 6.2 to about 6.8, about 6.4 to about 6.6, or about 6.5. In one embodiment, the formulation has a pH of between about 4.6 to about 7.6. In one embodiment, the formulation has a pH of between about 4.0 to about 8.0.

[0156] In one embodiment, the formulation comprises an active ingredient. The active ingredient can be any active ingredient to treat a disease or disorder in a subject in need thereof provided that the active ingredient can be administered via injection to the subject in need thereof. In one embodiment, the active ingredient is selected from a small molecule, a peptide fragment, a biologic, a nanoparticle, an antibody, an antibody fragment, or a small molecule antiviral.Methods of Treatment

[0157] In one aspect, the present disclosure provides a method of treating a disease or disorder in subject in need thereof, the method comprising administering to the subject via injection a formulation comprising a hyaluronidase enzyme and a therapeutically effective amount of an active ingredient. The formulation is described elsewhere herein. In one embodiment, the hyaluronidase enzyme is a recombinant human hyaluronidase PH20 enzyme. In one embodiment, the subject is a mammal. In one embodiment, the subject is a human. In one embodiment, the formulation is administered to the subject via a subcutaneous injection. In one embodiment, the formulation is self-administered. In another embodiment, the formulation is administered to the subject by a healthcare professional. In yet another embodiment, the formulation is administered to the subject by a layperson, such as acaregiver. In one embodiment, the formulation is administered using an autoinjector. In one embodiment, the formulation is administered using a HVAI. In one embodiment, the formulation is administered from a prefdled syringe using a HVAI. In another embodiment, the formulation is administered manually using a manually triggered injection device. In another embodiment, the formulation is administered from an on-body device.

[0158] In one embodiment, the formulation is administered to the abdomen or thigh of the subject. In one embodiment, the formulation is subcutaneously administered to the abdomen or thigh of the subject.

[0159] In one embodiment, the amount of the disclosed formulation administered to the subject is dependent on the subject being treated, the severity of the disorder or condition, the rate of administration, the disposition of the compounds, and / or the discretion of the prescribing physician. In one embodiment, an effective dosage of the active ingredient in the disclosed formulation is in the range of about 0.001 to about 100 mg per kg body weight per day, such as about 1 to about 35 mg / kg / day, in single or divided doses. For a 70 kg human, this would amount to about 0.05 to 7 g / day, such as about 0.05 to about 2.5 g / day. In some instances, dosage levels below the lower limit of the aforesaid range may be more than adequate, while in other cases still larger doses may be employed without causing any harmful side effect, for example by dividing such larger doses into several small doses for administration throughout the day.

[0160] In one embodiment, the disclosed formulation is administered to the subject in multiple doses. Dosing may be about once, twice, three times, four times, five times, six times, or more than six times per day. Dosing may be about once a month, once every two weeks, once a week, or once every other day. In one embodiment, the disclosed formulation is administered about once per day to about 6 times per day. In one embodiment, the administration of the disclosed formulation continues for less than about 7 days. In yet another embodiment the administration continues for more than about 6, 10, 14, 28 days, two months, six months, or one year. In some cases, continuous dosing is achieved and maintained as long as necessary.

[0161] In one embodiment, an effective dosage of the active ingredient in the disclosed formulation is in the range of about 1 mg / mL to about 500 mg / mL, about 10 mg / mL to about 450 mg / mL, about 20 mg / mL to about 400 mg / mL, about 30 mg / mL to about 350 mg / mL, about 50 mg / mL to about 300 mg / mL, about 75 mg / mL to about 250 mg / mL, about 100 mg / mL to about 200 mg / mL, or about 150 mg / mL.

[0162] In one embodiment, the formulation is administered using a HVAI fitted with a needle. In one embodiment, the needle is a 20 gauge needle, a 21 gauge needle, a 22 gauge needle, a 23 gauge needle,a 24 gauge needle, a 25 gauge needle, a 26 gauge needle, a 27 gauge needle, a 28 gauge needle, a 29 gauge needle, a 30 gauge needle, or a 31 gauge needle. In one embodiment, the needle is a 27 gauge regular wall needle x Vi a 27 gauge regular wall x 5 / 8” needle, a 27 gauge regular wall x 3 / 4” needle, a 27 gauge regular wall x 1” needle, a 27 gauge thin wall x needle, a 27 gauge thin wall x 5 / 8” needle, a 27 gauge thin wall x 3 / 4” needle, a 27 gauge thin wall x 1” needle, a 25 gauge regular wall x LT needle, a 25 gauge regular wall x 5 / 8” needle, a 25 gauge regular wall x 3 / 4” needle, a 25 gauge regular wall x 1” needle, a 25 gauge thin wall x ! ” needle, a 25 gauge thin wall x 5 / 8” needle, a 25 gauge thin wall x 3 / 4” needle, a 25 gauge thin wall x 1” needle, a 23 gauge regular wall x Vz needle, a 23 gauge regular wall x 5 / 8” needle, a 23 gauge regular wall x 3 / 4” needle, or a 23 gauge regular wall x 1” needle. In one embodiment, the needle is a 25 gauge thin wall xx / i needle. In one embodiment, the needle is a 25 gauge thin wall x 5 / 8” needle.

[0163] In one embodiment, a high volume of the formulation is administered to the subject. In one embodiment, a “high volume” is greater than 2.25 mL in a single administration. In one embodiment,3 mL to 5 mL, 3 mL to 10 mL, 3 mL to 15 mL, 3 mL to 20 mL, 3 mL to 25 mL, 3 ml to 30 mL, 3 mL to 35 mL, 3 mL to 40 mL, 3 mL to 45 mL, 3 mL to 50 mL, 5 mL to 10 mL, 5 mL to 15 mL, 5 mL to 20 mL, 5 mL to 25 mL, 5 mL to 30 mL, 5 mL to 35 mL, 5 mL to 40 mL; 5 mL to 45 mL, 5 mL to 50 mL, 10 mL to 15 mL; 10 mL to 20 mL; 10 mL to 25 mL; 10 mL to 30 mL; 10 mL to 35 mL; 10 mL to 40 mL, or 10 mL to 50 mL are administered to the subject in a single administration. In one embodiment, about 3 mL to about 5 mL, about 3 mL to about 10 mL, about 3 mL to about 15 mL, about 3 mL to about 20 mL, about 3 mL to about 25 mL, about 3 ml to about 30 mL, about 3 mL to about 35 mL, about 3 mL to about 40 mL, about 3 mL to about 45 mL, about 3 mL to about 50 mL, about 5 mL to about 10 mL, about 5 mL to about 15 mL, about 5 mL to about 20 mL, about 5 mL to about 25 mL, about 5 mL to about 30 mL, about 5 mL to about 35 mL, about 5 mL to about 40 mL; about 5 mL to about 45 mL, about 5 mL to about 50 mL, about 10 mL to about 15 mL; about 10 mL to about 20 mL; about 10 mL to about 25 mL; about 10 mL to about 30 mL; about 10 mL to about 35 mL; about 10 mL to about 40 mL, or about 10 mL to about 50 mL are administered to the subject in a single administration. In one embodiment, at least about 3 mL, at least about 3.5 mL, at least about4 mL, at least about 4.5 mL, at least about 5.5 mL, at least about 6 mL, at least about 6.5 mL, at least about 7 mL, at least about 7.5 mL, at least about 8 mL, at least about 8.5 mL, at least about 9 mL, at least about 9.5 mL, at least about 10 mL, at least about 10.5 mL, at least about 11 mL, at least about 11.5 mL, at least about 12 mL, at least about 12.5 mL, at least about 13 mL, at least about 13.5 mL, at least about 14 mL, at least about 14.5 mL, at least about 15 mL, at least about 15.5 mL, at least about 16 mL, at least about 16.5 mL, at least about 17 mL, at least about 17.5 mL, at least about 18mL, at least about 18.5 mL, at least about 19 m , at least about 19.5 mb, at least about 20 mb, at least about 25 mL, at least about 30 mL, at least about 35 mL, at least about 40 mL, at least about 45 mL, or at least about 50 mL are administered to the subject in a single administration. In one embodiment, at least 3 mL, at least 3.5 mL, at least 4 mL, at least 4.5 mL, at least 5.5 mL, at least 6 mL, at least 6.5 mL, at least 7 mL, at least 7.5 mL, at least 8 mL, at least 8.5 mL, at least 9 mL, at least 9.5 mL, at least 10 mL, at least 10.5 mL, at least 11 mL, at least 11.5 mL, at least 12 mL, at least 12.5 mL, at least 13 mL, at least 13.5 mL, at least 14 mL, at least 14.5 mL, at least 15 mL, at least 15.5 mL, at least 16 mL, at least 16.5 mL, at least 17 mL, at least 17.5 mL, at least 18 mL, at least 18.5 mL, at least 19 mL, at least 19.5 mL, at least 20 mL, at least 25 mL, at least 30 mL, at least 35 mL, at least40 mL, at least 45 mL, or at least 50 mL are administered to the subject in a single administration.

[0164] In one embodiment, greater than about 2.25 mL, about 2.5 mL, about 3.0 mL, about 3.5 mL, about 4.0 mL, about 4.5 mL, about 5.0 mL, about 5.5 mL, about 6.0 mL, about 6.5 mL, about 7.0 mL, about 7.5 mL, about 8.0 mL, about 8.5 mL, about 9.5 mL, about 10 mL, about 12 mL, about 14 mL, about 16 mL, about 18 mL, or about 20 mL of the formulation is administered to the subject in a single administration. In one embodiment, greater than about 2.5 mL, about 3.0 mL, about 3.5 mL, about 4.0 mL, about 4.5 mL, about 5.0 mL, about 5.5 mL, about 6.0 mL, about 6.5 mL, about 7.0 mL, about 7.5 mL, about 8.0 mL, about 8.5 mL, about 9.5 mL, about 10 mL, about 12 mL, about 14 mL, about 16 mL, about 18 mL, or about 20 mL of the formulation is administered to the subject in about 10 seconds, about 12 seconds, about 16 seconds, about 18 seconds, about 20 seconds, about 22 seconds, about 24 seconds, about 26 seconds, about 28 seconds, about 30 seconds, about 32 seconds, about 34 seconds, about 36 seconds, about 38 seconds, about 40 seconds, about 42 seconds, about 44 seconds, about 46 seconds, about 48 seconds, about 50 seconds, about 52 seconds, about 54 seconds, about 56 seconds, about 58 seconds, about 60 seconds, about 65 seconds, about 70 seconds, about 75 seconds, about 80 seconds, about 85 seconds, about 90 seconds, about 95 seconds, about 100 seconds, about 105 seconds, about 110 seconds, about 115 seconds, or about 120 seconds.

[0165] In one embodiment, about 5 mL of the formulation can be administered to the subject in about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, or about 60 seconds.

[0166] In one embodiment, about 10 mL of the formulation can be administered to the subject in about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, or about 60 seconds.

[0167] In one embodiment, about 10.5 mL of the formulation can be administered to the subject in about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, or about 60 seconds.

[0168] In some embodiments, the speed at which a high volume of the formulation can be administered to the subject depends on the gauge of the needle used to inject the formulation. In one embodiment, the formulation is administered to the subject using an autoinjector and a 23 gauge needle. In one embodiment, about 10 mL of the formulation can be administered to the subject in between about 5 seconds to about 45 seconds, about 5 seconds to about 40 seconds, about 10 seconds to about 40 seconds, about 10 seconds to about 35 seconds, about 15 seconds to about 35 seconds, about 15 seconds to about 30 seconds, about 20 seconds to 30 seconds, about 15 seconds to about 25 seconds, or about 20 seconds using an autoinjector and a 23 gauge needle. In one embodiment, about 5.5 mL of the formulation can be administered to the subject in between about 5 seconds to about 45 seconds, about 5 seconds to about 40 seconds, about 10 seconds to about 40 seconds, about 10 seconds to about 35 seconds, about 15 seconds to about 35 seconds, about 15 seconds to about 30 seconds, about 15 seconds to about 25 seconds, or about 20 seconds using an autoinjector and a 23 gauge needle. In one embodiment, about 5.5 mL of the formulation can be administered to the subject in between about 15 seconds to about 60 seconds, about 20 seconds to about 60 seconds, about 25 seconds to about 60 seconds, about 25 seconds to about 55 seconds, about 30 seconds to about 55 seconds, about 30 seconds to about 50 seconds, about 35 seconds to about 50 seconds, about 40 seconds to about 50 seconds, or about 45 seconds using an autoinjector and a 23 gauge needle. In another embodiment, the formulation is administered to the subject using an autoinjector and a 25 gauge needle. In one embodiment, about 10 mL of the formulation can be administered to the subject in between about 15 seconds to about 60 seconds, about 20 seconds to about 60 seconds, about 25 seconds to about 60 seconds, about 25 seconds to about 55 seconds, about 30 seconds to about 55 seconds, about 30 seconds to about 50 seconds, about 35 seconds to about 50 seconds, about 40 seconds to about 50 seconds, or about 45 seconds using an autoinjector and a 25 gauge needle. In one embodiment, about 5.5 mL of the formulation can be administered to the subject in between about 5 seconds to about 45 seconds, about 5 seconds to about 40 seconds, about 10 seconds to about 40 seconds, about 10 seconds to about 35 seconds, about 15 seconds to about 35 seconds, about 15 seconds to about 30 seconds, about 15 seconds to about 25 seconds, or about 20 seconds using an autoinjector and a 23 gauge needle. In one embodiment, about 5.5 mL of the formulation can be administered to the subject in between about 15 seconds to about 60 seconds, about 20 seconds toabout 60 seconds, about 25 seconds to about 60 seconds, about 25 seconds to about 55 seconds, about 30 seconds to about 55 seconds, about 30 seconds to about 50 seconds, about 35 seconds to about 50 seconds, about 40 seconds to about 50 seconds, or about 45 seconds using an autoinjector and a 25 gauge needle. In another embodiment, the formulation is administered to the subject using an autoinjector and a 27 gauge needle. In one embodiment, about 10 mL of the formulation can be administered to the subject in between about 15 seconds to about 60 seconds, about 20 seconds to about 60 seconds, about 25 seconds to about 60 seconds, about 25 seconds to about 55 seconds, about 30 seconds to about 55 seconds, about 30 seconds to about 50 seconds, about 35 seconds to about 50 seconds, about 40 seconds to about 50 seconds, or about 45 seconds using an autoinjector and a 27 gauge needle. In one embodiment, about 5.5 mL of the formulation can be administered to the subject in between about 15 seconds to about 60 seconds, about 20 seconds to about 60 seconds, about 25 seconds to about 60 seconds, about 25 seconds to about 55 seconds, about 30 seconds to about 55 seconds, about 30 seconds to about 50 seconds, about 35 seconds to about 50 seconds, about 40 seconds to about 50 seconds, or about 45 seconds using an autoinjector and a 27 gauge needle.

[0169] In one embodiment, using a 23 gauge needle with a HVAI and the disclosed formulation provides about 40%, about 42%, about 44%, about 46%, about 48%, about 50%, about 52%, about 54%, about 56%, about 58%>, or about 60%> faster injection times than using a 25 gauge needle with a HVAI.

[0170] In one embodiment, the device has a variable delivery rate that can deliver the formulation faster at the start of the inj ection and slower at the completion of the inj ection. In another embodiment, the device has a variable delivery rate that delivers the formulation slower at the initiation of the injection and faster at the completion of the injection.

[0171] In one embodiment, the viscosity and volume of the disclosed formulation affect the time needed to inject the formulation into a subject. However, the full volume of the formulation can be delivered from a HVAI at a rate of approximately 0.08-1.0 mL / sec. For example, this would provide target delivery rate ranges of 10-120 seconds for a 10 mL dose volume. The HVAI may deliver 10 mL of the formulation at a rate of 0.33 mL / sec. In one embodiment, HVAI delivers the full deliverable volume of the formulation at a rate of: 0.5 mL / 10 sec., 0.75 mL / 10 sec., 1 mL / 10 sec., 1.25 mL / 10 sec., 1.5 mL / 10 sec., 1.75 mL / 10 sec, 2 mL / 10 sec., 2.25 mL / 10 sec, 2.5 mL / 10 sec., 2.75 mL / 10 sec, 3 mL / 10 sec., 3.25 mL / 10 sec, 3.5 mL / 10 sec., 3.75 mL / 10 sec, 4 mL / 10 sec., 4.25 mL / 10 sec, 4.5 mL / 10 sec., 4.75 mL / 10 sec, or about 5 mL / 10 sec. In one embodiment, the HVAI delivers the full deliverable volume of the formulation at a rate of: 2 mL / 30 sec., 2.5 mL / 30 sec., 3 mL / 30 sec., 3.5 mL / 30 sec., 4 mL / 30 sec., 4.5 mL / 30 sec., 5 mL / 30 sec., 5.5 mL / 30 sec., 6 mL / 30 sec., 6.5 mL / 30sec., 7 mL / 30 sec., 7.5 mL / 30 sec., 8 mL / 30 sec., 8.5 mL / 30 sec., 9 mL / 30 sec., 9.5 mL / 30 sec., 10 mL / 30 sec., or 10.5 mL / 30 sec. In one embodiment, delivers the full deliverable volume of the formulation at a rate of: 4 mL / min, 5 mL / min, 6 mL / rnin, 7 mL / min, 8 mL / min, 9 mL / min, 10 mL / min, 11 mL / min, 12 mL / min, 13 mL / min, 14 mL / min, 15 mL / min, 16 mL / min, 17 mL / min, 18 mL / min, 19 mL / min, 20 mL / min, 21 mL / min.

[0172] In one embodiment, the disclosed formulation is administered to the subject at a rate of about 0.05 mL / sec, about 0.06 mL / sec, about 0.07 mL / sec, about 0.08 mL / sec, about 0.09 mL / sec, about0.10 mL / sec, about 0.11 mL / sec, about 0.12 mL / sec, about 0.13 mL / sec, about 0.14 mL / sec, about0.15 mL / sec, about 0.16 mL / sec, about 0.17 mL / sec, about 0.18 mL / sec, about 0.19 mL / sec, about0.20 mL / sec, about 0.21 mL / sec, about 0.22 mL / sec, about 0.23 mL / sec, about 0.24 mL / sec, about0.25 mL / sec, about 0.26 mL / sec, about 0.27 mL / sec, about 0.28 mL / sec, about 0.29 mL / sec, about0.30 mL / sec, about 0.31 mL / sec, about 0.32 mL / sec, about 0.33 mL / sec, about 0.34 mL / sec, about0.35 mL / sec, about 0.36 mL / sec, about 0.37 mL / sec, about 0.38 mL / sec, about 0.39 mL / sec, about 0.4 mL / sec, about 0.41 mL / sec, about 0.42 mL / sec, about 0.43 mL / sec, about 0.44 mL / sec, about 0.45 mL / sec, about 0.46 mL / sec, about 0.47 mL / sec, about 0.48 mL / sec, about 0.49 mL / sec, about 0.50 mL / sec, about 0.51 mL / sec, about 0.52 mL / sec, about 0.53 mL / sec, about 0.54 mL / sec, about 0.55 mL / sec, about 0.56 mL / sec, about 0.57 mL / sec, about 0.58 mL / sec, about 0.59 mL / sec, about 0.60 mL / sec, about 0.61 mL / sec, about 0.62 mL / sec, about 0.63 mL / sec, about 0.64 mL / sec, about 0.65 mL / sec, about 0.66 mL / sec, about 0.67 mL / sec, 0.68 mL / sec, about 0.69 mL / sec, about 0.70 mL / sec, about 0.71 mL / sec, about 0.72 mL / sec, about 0.73 mL / sec, about 0.74 mL / sec, about 0.75 mL / sec, about 0.76 mL / sec, about 0.77 mL / sec, about 0.78 mL / sec, about 0.79 mL / sec, about 0.80 mL / sec, about 0.81 mL / sec, about 0.82 mL / sec, about 0.83 mL / sec, about 0.84 mL / sec, about 0.85 mL / sec, about 0.86 mL / sec, about 0.87 mL / sec, about 0.88 mL / sec, about 0.89 mL / sec, about 0.90 mL / sec, about 0.91 mL / sec, about 0.92 mL / sec, about 0.93 mL / sec, about 0.94 mL / sec, about 0.95 mL / sec, about 0.96 mL / sec, about 0.97 mL / sec, about 0.98 mL / sec, about 0.99 mL / sec, or about 1.0 mL / sec.

[0173] In one embodiment, the disclosed formulation is administered to the subject, using a prefilled syringe fitted with a needle having a gauge between 20 and 33, at a rate of about 0.05 mL / sec, about 0.06 mL / sec, about 0.07 mL / sec, about 0.08 mL / sec, about 0.09 mL / sec, about 0.10 mL / sec, about0.11 mL / sec, about 0.12 mL / sec, about 0.13 mL / sec, about 0.14 mL / sec, about 0.15 mL / sec, about0.16 mL / sec, about 0.17 mL / sec, about 0.18 mL / sec, about 0.19 mL / sec, about 0.20 mL / sec, about0.21 mL / sec, about 0.22 mL / sec, about 0.23 mL / sec, about 0.24 mL / sec, about 0.25 mL / sec, about0.26 mL / sec, about 0.27 mL / sec, about 0.28 mL / sec, about 0.29 mL / sec, about 0.30 mL / sec, about0.31 mL / sec, about 0.32 mL / sec, about 0.33 mL / sec, about 0.34 mL / sec, about 0.35 mL / sec, about0.36 mL / sec, about 0.37 mL / sec, about 0.38 mL / sec, about 0.39 mL / sec, about 0.4 mL / sec, about 0.41 mL / sec, about 0.42 mL / sec, about 0.43 mL / sec, about 0.44 mL / sec, about 0.45 mL / sec, about 0.46 mL / sec, about 0.47 mL / sec, about 0.48 mL / sec, about 0.49 mL / sec, about 0.50 mL / sec, about 0.51 mL / sec, about 0.52 mL / sec, about 0.53 mL / sec, about 0.54 mL / sec, about 0.55 mL / sec, about 0.56 mL / sec, about 0.57 mL / sec, about 0.58 mL / sec, about 0.59 mL / sec, about 0.60 mL / sec, about 0.61 mL / sec, about 0.62 mL / sec, about 0.63 mL / sec, about 0.64 mL / sec, about 0.65 mL / sec, about 0.66 mL / sec, about 0.67 mL / sec, 0.68 mL / sec, about 0.69 mL / sec, about 0.70 mL / sec, about 0.71 mL / sec, about 0.72 mL / sec, about 0.73 mL / sec, about 0.74 mL / sec, about 0.75 mL / sec, about 0.76 mL / sec, about 0.77 mL / sec, about 0.78 mL / sec, about 0.79 mL / sec, about 0.80 mL / sec, about 0.81 mL / sec, about 0.82 mL / sec, about 0.83 mL / sec, about 0.84 mL / sec, about 0.85 mL / sec, about 0.86 mL / sec, about 0.87 mL / sec, about 0.88 mL / sec, about 0.89 mL / sec, about 0.90 mL / sec, about 0.91 mL / sec, about 0.92 mL / sec, about 0.93 mL / sec, about 0.94 mL / sec, about 0.95 mL / sec, about 0.96 mL / sec, about 0.97 mL / sec, about 0.98 mL / sec, about 0.99 mL / sec, or about 1.0 mL / sec.

[0174] In one embodiment, the disclosed formulation is administered to the subject from a high volume autoinjector disclosed herein at a rate of about 0.05 mL / sec, about 0.06 mL / sec, about 0.07 mL / sec, about 0.08 mL / sec, about 0.09 mL / sec, about 0.10 mL / sec, about 0.11 mL / sec, about 0.12 mL / sec, about 0.13 mL / sec, about 0.14 mL / sec, about 0.15 mL / sec, about 0.16 mL / sec, about 0.17 mL / sec, about 0.18 mL / sec, about 0.19 mL / sec, about 0.20 mL / sec, about 0.21 mL / sec, about 0.22 mL / sec, about 0.23 mL / sec, about 0.24 mL / sec, about 0.25 mL / sec, about 0.26 mL / sec, about 0.27 mL / sec, about 0.28 mL / sec, about 0.29 mL / sec, about 0.30 mL / sec, about 0.31 mL / sec, about 0.32 mL / sec, about 0.33 mL / sec, about 0.34 mL / sec, about 0.35 mL / sec, about 0.36 mL / sec, about 0.37 mL / sec, about 0.38 mL / sec, about 0.39 mL / sec, about 0.4 mL / sec, about 0.41 mL / sec, about 0.42 mL / sec, about 0.43 mL / sec, about 0.44 mL / sec, about 0.45 mL / sec, about 0.46 mL / sec, about 0.47 mL / sec, about 0.48 mL / sec, about 0.49 mL / sec, about 0.50 mL / sec, about 0.51 mL / sec, about 0.52 mL / sec, about 0.53 mL / sec, about 0.54 mL / sec, about 0.55 mL / sec, about 0.56 mL / sec, about 0.57 mL / sec, about 0.58 mL / sec, about 0.59 mL / sec, about 0.60 mL / sec, about 0.61 mL / sec, about 0.62 mL / sec, about 0.63 mL / sec, about 0.64 mL / sec, about 0.65 mL / sec, about 0.66 mL / sec, about 0.67 mL / sec, 0.68 mL / sec, about 0.69 mL / sec, about 0.70 mL / sec, about 0.71 mL / sec, about 0.72 mL / sec, about 0.73 mL / sec, about 0.74 mL / sec, about 0.75 mL / sec, about 0.76 mL / sec, about 0.77 mL / sec, about 0.78 mL / sec, about 0.79 mL / sec, about 0.80 mL / sec, about 0.81 mL / sec, about 0.82 mL / sec, about 0.83 mL / sec, about 0.84 mL / sec, about 0.85 mL / sec, about 0.86 mL / sec, about 0.87 mL / sec, about 0.88 mL / sec, about 0.89 mL / sec, about 0.90 mL / sec, about 0.91 mL / sec, about 0.92 mL / sec,about 0.93 mL / sec, about 0.94 mL / sec, about 0.95 mL / sec, about 0.96 mL / sec, about 0.97 mL / sec, about 0.98 mL / sec, about 0.99 mL / sec, or about 1.0 mL / sec.

[0175] In one embodiment, the disclosed formulation is contained in a prefilled syringe fitted with a needle having a gauge between 20 and 33 wherein the prefilled syringe is contained in a high volume autoinjector disclosed herein and the formulation is administered to the subject at a rate of about 0.05 mL / sec, about 0.06 mL / sec, about 0.07 mL / sec, about 0.08 mL / sec, about 0.09 mL / sec, about 0.10 mL / sec, about 0.11 mL / sec, about 0.12 mL / sec, about 0.13 mL / sec, about 0.14 mL / sec, about 0.15 mL / sec, about 0.16 mL / sec, about 0.17 mL / sec, about 0.18 mL / sec, about 0.19 mL / sec, about 0.20 mL / sec, about 0.21 mL / sec, about 0.22 mL / sec, about 0.23 mL / sec, about 0.24 mL / sec, about 0.25 mL / sec, about 0.26 mL / sec, about 0.27 mL / sec, about 0.28 mL / sec, about 0.29 mL / sec, about 0.30 mL / sec, about 0.31 mL / sec, about 0.32 mL / sec, about 0.33 mL / sec, about 0.34 mL / sec, about 0.35 mL / sec, about 0.36 mL / sec, about 0.37 mL / sec, about 0.38 mL / sec, about 0.39 mL / sec, about 0.4 mL / sec, about 0.41 mL / sec, about 0.42 mL / sec, about 0.43 mL / sec, about 0.44 mL / sec, about 0.45 mL / sec, about 0.46 mL / sec, about 0.47 mL / sec, about 0.48 mL / sec, about 0.49 mL / sec, about 0.50 mL / sec, about 0.51 mL / sec, about 0.52 mL / sec, about 0.53 mL / sec, about 0.54 mL / sec, about 0.55 mL / sec, about 0.56 mL / sec, about 0.57 mL / sec, about 0.58 mL / sec, about 0.59 mL / sec, about 0.60 mL / sec, about 0.61 mL / sec, about 0.62 mL / sec, about 0.63 mL / sec, about 0.64 mL / sec, about 0.65 mL / sec, about 0.66 mL / sec, about 0.67 mL / sec, 0.68 mL / sec, about 0.69 mL / sec, about 0.70 mL / sec, about 0.71 mL / sec, about 0.72 mL / sec, about 0.73 mL / sec, about 0.74 mL / sec, about 0.75 mL / sec, about 0.76 mL / sec, about 0.77 mL / sec, about 0.78 mL / sec, about 0.79 mL / sec, about 0.80 mL / sec, about 0.81 mL / sec, about 0.82 mL / sec, about 0.83 mL / sec, about 0.84 mL / sec, about 0.85 mL / sec, about 0.86 mL / sec, about 0.87 mL / sec, about 0.88 mL / sec, about 0.89 mL / sec, about 0.90 mL / sec, about 0.91 mL / sec, about 0.92 mL / sec, about 0.93 mL / sec, about 0.94 mL / sec, about 0.95 mL / sec, about 0.96 mL / sec, about 0.97 mL / sec, about 0.98 mL / sec, about 0.99 mL / sec, or about 1.0 mL / sec.

[0176] In one embodiment, about 3 mL to about 50 mL of the formulation is administered to a subject via subcutaneous administration, wherein the subcutaneous administration occurs at a rate of about 0.10 mL / sec to about 1.0 mL / sec. In one embodiment, about 3 mL to about 15 mL of the formulation is administered to the subject via subcutaneous administration, wherein the subcutaneous administration occurs at a rate of about 0.10 mL / sec to about 1.0 mL / sec. In one embodiment, the formulation is administered subcutaneously to the subject from a prefilled syringe. In one embodiment, the prefilled syringe comprises a needle having a gauge of about 20 to about 31. In one embodiment, the prefilled syringe comprises a needle having a gauge of about 23, about 25, or about27. In one embodiment, the formulation is administered to the subject from the prefilled syringe using a HVAI described elsewhere herein.

[0177] In one embodiment, about 5 mb of the formulation is administered to the subject at a rate of about 0.14 mL / sec to about 0.21 mL / sec. In one embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.14 mL / sec to about 0.21 mL / sec from a prefilled syringe using a HVAI. In one embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.28 mL / sec to about 0.42 mL / sec. In one embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.28 mL / sec to about 0.42 mL / sec from a prefilled syringe using a HVAI. In one embodiment, about 10 mL of the formulation is administered to the subject at a rate of about 0.32 mL / sec to about 0.42 mL / sec. In one embodiment, about 10 mL of the formulation is administered to the subject at a rate of about 0.32 mL / sec to about 0.42 mL / sec from a prefilled syringe using a HVAI. In one embodiment, about 10.5 mL of the formulation is administered to the subject at a rate of about 0.40 mL / sec to about 1.0 mL / sec. In one embodiment, about 10.5 mL of the formulation is administered to the subject at a rate of about 0.40 mL / sec to about 1.0 mL / sec from a prefilled syringe using a HVAI. In one embodiment, about 10.5 mL of the formulation is administered to the subject at a rate of at least 0.7 mL / sec. In one embodiment, about 10.5 mL of the formulation is administered to the subject at a rate of at least 0.7 mL / sec from a prefilled syringe using a HVAI.

[0178] In one embodiment, about 5 mL, about 5.5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, about 10 mL, about 10.5 mL, about 11 mL, about 12 mL, about 13 mL, about 14 mL, or about 15 mL of the disclosed formulation can be administered to a subject in 15 seconds or less using a needle having a gauge of 20 to 31 . In one embodiment, about 5 mL, about 5.5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, about 10 mL, about 10.5 mL, about 11 mL, about 12 mL, about 13 mL, about 14 mL, or about 15 mL of the disclosed formulation can be administered to a subject in 15 seconds or less using a 27 gauge needle. In one embodiment, about 10.5 mL of the disclosed formulation can be administered to a subject in 15 seconds or less using a 27 gauge needle. In one embodiment, the needle is connected to a syringe wherein the syringe is prefilled with the disclosed formulation. In one embodiment, the prefilled syringe is contained in an autoinjector. In one embodiment, the prefilled syringe is contained in the HVAI described elsewhere herein.

[0179] In one embodiment, about 3 mL to about 50 mL of the formulation disclosed herein is administered to the subject with a starting delivery force of about 1 Ibf to about 200 Ibf, about 1 Ibf to about 190 Ibf, about 1 Ibf to about 180 Ibf, about 1 Ibf to about 170 Ibf, about 1 Ibf to about 160 Ibf, about 1 Ibf to about 150 Ibf, about 1 Ibf to about 140 Ibf, about 1 Ibf to about 130 Ibf, about 1 Ibf toabout 120 Ibf, about 1 Ibf to about 110 Ibf, 1 Ibf to about 100 Ibf, 1 Ibf to about 100 Ibf, about 1 Ibf to about 90 Ibf, about 1 Ibf to about 80 Ibf, about 1 Ibf to about 70 Ibf, about 1 Ibf to about 60 Ibf, about 1 Ibf to about 50 Ibf, about 1 Ibf to about 40 Ibf, about 1 Ibf to about 30 Ibf, or about 25 Ibf from a prefilled syringe using a HVAI. In one embodiment, about 3 mL to about 50 mL of the formulation disclosed herein is administered to the subject with a starting delivery force of about 1 Ibf, about 2 Ibf, about 3 Ibf, about 4 Ibf, 5 Ibf, about 6 Ibf, about 7 Ibf, about 8 Ibf, about 9 Ibf, about 10 Ibf, about 11 Ibf, about 12 Ibf, about 13 Ibf, about 14 Ibf, about 15 Ibf, about 16 Ibf, about 17 Ibf, about 18 Ibf, about 19 Ibf, about 20 Ibf, about 21 Ibf, about 22 Ibf, about 23 Ibf, about 24 Ibf, about 25 Ibf, about 26 Ibf, about 27 Ibf, about 28 Ibf, about 29 Ibf, about 30 Ibf, about 31 Ibf, about 32 Ibf, about 33 Ibf, about 34 Ibf, about 35 Ibf, about 36 Ibf, about 37 Ibf, about 38 Ibf, about 39 Ibf, about 40 Ibf, about 41 Ibf, about 42 Ibf, about 43 Ibf, about 44 Ibf, about 45 Ibf, about 46 Ibf, about 47 Ibf, about 48 Ibf, about 49 Ibf, about 50 Ibf, about 51 Ibf, about 52 Ibf, about 53 Ibf, about 54 Ibf, about 55 Ibf, about 56 Ibf, about 57 Ibf, about 58 Ibf, about 59 Ibf, about 60 Ibf, about 61 Ibf, about 62 Ibf, about 63 Ibf, about 64 Ibf, about 65 Ibf, about 66 Ibf, about 67 Ibf, about 68 Ibf, about 69 Ibf, about 70 Ibf, about 71 Ibf, about 72 Ibf, about 73 Ibf, about 74 Ibf, about 75 Ibf, about 76 Ibf, about 77 Ibf, about 78 Ibf, about 79 Ibf, about 80 Ibf, about 81 Ibf, about 82 Ibf, about 83 Ibf, about 84 Ibf, about 85 Ibf, about 86 Ibf, about 87 Ibf, about 88 Ibf, about 89 Ibf, about 90 Ibf, about 91 Ibf, about 92 Ibf, about 93 Ibf, about 94 Ibf, about 95 Ibf, about 96 Ibf, about 97 Ibf, about 98 Ibf, about 99 Ibf, about 100 Ibf, about 101 Ibf, about 102 Ibf, about 103 Ibf, about 104 Ibf, about 105 Ibf, about 106 Ibf, about 107 Ibf, about 108 Ibf, about 109 Ibf, about110 Ibf, about 111 Ibf, about 112 Ibf, about 113 Ibf, about 114 Ibf, about 115 Ibf, about 116 Ibf, about117 Ibf, about 118 Ibf, about 119 Ibf, about 120 Ibf, about 121 Ibf, about 122 Ibf, about 123 Ibf, about124 Ibf, about 125 Ibf, about 126 Ibf, about 127 Ibf, about 128 Ibf, about 129 Ibf, about 130 Ibf, about131 Ibf, about 132 Ibf, about 133 Ibf, about 134 Ibf, about 135 Ibf, about 136 Ibf, about 137 Ibf, about138 Ibf, about 139 Ibf, about 140 Ibf, about 141 Ibf, about 142 Ibf, about 143 Ibf, about 144 Ibf, about145 Ibf, about 146 Ibf, about 147 Ibf, about 148 Ibf, about 149 Ibf, about 150 Ibf, about 151 Ibf, about152 Ibf, about 153 Ibf, about 154 Ibf, about 155 Ibf, about 156 Ibf, about 157 Ibf, about 158 Ibf, about159 Ibf, about 160 Ibf, about 161 Ibf, about 162 Ibf, about 163 Ibf, about 164 Ibf, about 165 Ibf, about166 Ibf, about 167 Ibf, about 168 Ibf, about 169 Ibf, about 170 Ibf, about 171 Ibf, about 172 Ibf, about173 Ibf, about 174 Ibf, about 175 Ibf, about 176 Ibf, about 177 Ibf, about 178 Ibf, about 179 Ibf, about180 Ibf, about 181 Ibf, about 182 Ibf, about 183 Ibf, about 184 Ibf, about 185 Ibf, about 186 Ibf, about187 Ibf, about 188 Ibf, about 189 Ibf, about 190 Ibf, about 191 Ibf, about 192 Ibf, about 193 Ibf, about194 Ibf, about 195 Ibf, about 196 Ibf, about 197 Ibf, about 198 Ibf, about 199 Ibf, or about 200 Ibf from a prefilled syringe using a HVAI.

[0180] In one embodiment, the starting delivery force refers to the starting delivery force exerted on the subject’s tissue upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the starting delivery force refers to the starting delivery force exerted the syringe stopper of the prefilled syringe in a HVAI as the disclosed formulation is delivered to a subject. In one embodiment, the starting delivery force refers to the starting delivery force exerted on the formulation upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the starting delivery force refers to the starting delivery force exerted on the prefilled syringe upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the starting delivery force refers to the starting delivery force generated by the HVAI device upon administration of the formulation from a prefilled syringe using a HVAI.

[0181] In one embodiment, about 3 mL to about 50 mL of the formulation disclosed herein is administered to the subject with an ending delivery force of about 1 Ibf to about 200 Ibf, about 1 Ibf to about 190 Ibf, about 1 Ibf to about 180 Ibf, about 1 Ibf to about 170 Ibf, about 1 Ibf to about 160 Ibf, about 1 Ibf to about 150 Ibf, about 1 Ibf to about 140 Ibf, about 1 Ibf to about 130 Ibf, about 1 Ibf to about 120 Ibf, about 1 Ibf to about 110 Ibf, 1 Ibf to about 100 Ibf, 1 Ibf to about 100 Ibf, about 1 Ibf to about 90 Ibf, about 1 Ibf to about 80 Ibf, about 1 Ibf to about 70 Ibf, about 1 Ibf to about 60 Ibf, about 1 Ibf to about 50 Ibf, about 1 Ibf to about 40 Ibf, about 1 Ibf to about 30 Ibf, about 5 Ibf to about30 Ibf, or about 5 Ibf to about 20 Ibf from a prefilled syringe using a HVAI. In one embodiment, about 3 mL to about 50 mL of the formulation disclosed herein is administered to the subject with an ending delivery force of about 1 Ibf, about 2 Ibf, about 3 Ibf, about 4 Ibf, 5 Ibf, about 6 Ibf, about 7 Ibf, about 8 Ibf, about 9 Ibf, about 10 Ibf, about 11 Ibf, about 12 Ibf, about 13 Ibf, about 14 Ibf, about 15 Ibf, about 16 Ibf, about 17 Ibf, about 18 Ibf, about 19 Ibf, about 20 Ibf, about 21 Ibf, about 22 Ibf, about 23 Ibf, about 24 Ibf, about 25 Ibf, about 26 Ibf, about 27 Ibf, about 28 Ibf, about 29 Ibf, about 30 Ibf, about31 Ibf, about 32 Ibf, about 33 Ibf, about 34 Ibf, about 35 Ibf, about 36 Ibf, about 37 Ibf, about 38 Ibf, about 39 Ibf, about 40 Ibf, about 41 Ibf, about 42 Ibf, about 43 Ibf, about 44 Ibf, about 45 Ibf, about 46 Ibf, about 47 Ibf, about 48 Ibf, about 49 Ibf, about 50 Ibf, about 51 Ibf, about 52 Ibf, about 53 Ibf, about 54 Ibf, about 55 Ibf, about 56 Ibf, about 57 Ibf, about 58 Ibf, about 59 Ibf, about 60 Ibf, about 61 Ibf, about 62 Ibf, about 63 Ibf, about 64 Ibf, about 65 Ibf, about 66 Ibf, about 67 Ibf, about 68 Ibf, about 69 Ibf, about 70 Ibf, about 71 Ibf, about 72 Ibf, about 73 Ibf, about 74 Ibf, about 75 Ibf, about 76 Ibf, about 77 Ibf, about 78 Ibf, about 79 Ibf, about 80 Ibf, about 81 Ibf, about 82 Ibf, about 83 Ibf, about 84 Ibf, about 85 Ibf, about 86 Ibf, about 87 Ibf, about 88 Ibf, about 89 Ibf, about 90 Ibf, about 91 Ibf, about 92 Ibf, about 93 Ibf, about 94 Ibf, about 95 Ibf, about 96 Ibf, about 97 Ibf, about 98 Ibf, about 99 Ibf, about 100 Ibf, about 101 Ibf, about 102 Ibf, about 103 Ibf, about 104 Ibf, about 105 Ibf, about 106 Ibf, about107 Ibf, about 108 Ibf, about 109 Ibf, about 110 Ibf, about 111 Ibf, about 112 Ibf, about 113 Ibf, about114 Ibf, about 115 Ibf, about 116 Ibf, about 117 Ibf, about 118 Ibf, about 119 Ibf, about 120 Ibf, about121 Ibf, about 122 Ibf, about 123 Ibf, about 124 Ibf, about 125 Ibf, about 126 Ibf, about 127 Ibf, about128 Ibf, about 129 Ibf, about 130 Ibf, about 131 Ibf, about 132 Ibf, about 133 Ibf, about 134 Ibf, about135 Ibf, about 136 Ibf, about 137 Ibf, about 138 Ibf, about 139 Ibf, about 140 Ibf, about 141 Ibf, about142 Ibf, about 143 Ibf, about 144 Ibf, about 145 Ibf, about 146 Ibf, about 147 Ibf, about 148 Ibf, about149 Ibf, about 150 Ibf, about 151 Ibf, about 152 Ibf, about 153 Ibf, about 154 Ibf, about 155 Ibf, about156 Ibf, about 157 Ibf, about 158 Ibf, about 159 Ibf, about 160 Ibf, about 161 Ibf, about 162 Ibf, about163 Ibf, about 164 Ibf, about 165 Ibf, about 166 Ibf, about 167 Ibf, about 168 Ibf, about 169 Ibf, about170 Ibf, about 171 Ibf, about 172 Ibf, about 173 Ibf, about 174 Ibf, about 175 Ibf, about 176 Ibf, about177 Ibf, about 178 Ibf, about 179 Ibf, about 180 Ibf, about 181 Ibf, about 182 Ibf, about 183 Ibf, about184 Ibf, about 185 Ibf, about 186 Ibf, about 187 Ibf, about 188 Ibf, about 189 Ibf, about 190 Ibf, about191 Ibf, about 192 Ibf, about 193 Ibf, about 194 Ibf, about 195 Ibf, about 196 Ibf, about 197 Ibf, about198 Ibf, about 199 Ibf, or about 200 Ibf from a prefilled syringe using a HVAI.

[0182] In one embodiment, the ending delivery force refers to the ending delivery force exerted on the subject’s tissue upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the ending delivery force refers to the ending delivery force exerted the syringe stopper of the prefilled syringe in a HVAI as the disclosed formulation is delivered to a subject. In one embodiment, the ending delivery force refers to the ending delivery force exerted on the formulation upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the ending delivery force refers to the ending delivery force exerted on the prefilled syringe upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the ending delivery force refers to the ending delivery force generated by the HVAI device upon administration of the formulation from a prefilled syringe using a HVAI.

[0183] In one embodiment, about 3 mb to about 50 mL of the formulation disclosed herein is administered to the subject with a starting pressure of about 10 psi to about 500 psi, about 10 psi to about 475 psi, about 10 psi to about 450 psi, about 10 psi to about 400 psi, about 10 psi to about 375 psi, about 10 psi to about 350 psi, about 10 psi to about 325 psi, about 10 psi to about 300 psi, about 20 psi to about 300 psi, about 20 psi to about 275 psi, about 30 psi to about 275 psi, about 30 psi to about 250 psi, about 40 psi to about 250 psi, about 40 psi to about 225 psi, about 50 psi to about 225 psi, or about 50 psi to about 200 psi from a prefilled syringe using a HVAI. In one embodiment, about 3 mL to about 50 mL of the formulation disclosed herein is administered to the subject with a starting pressure of about 50 psi, about 55 psi, about 60 psi, about 65 psi, about 70 psi, about 75 psi, about 80psi, about 85 psi, about 90 psi, about 95 psi, about 100 psi, about 105 psi, about 110 psi, about 115 psi, about 120 psi, about 125 psi, about 130 psi, about 135 psi, about 140 psi, about 145 psi, about 150 psi, about 155 psi, about 160 psi, about 165 psi, about 170 psi, about 175 psi, about 180 psi, about 185 psi, about 190 psi, about 195 psi, or about 200 psi from a prefilled syringe using a HVAI. In one embodiment, about 3 mb to about 50 mb of the formulation disclosed herein is administered to the subject with a starting pressure of about 50 psi, about 51 psi, about 52 psi, about 53 psi, about 54 psi, about 55 psi, about 56 psi, about 57 psi, about 58 psi, about 59 psi, about 60 psi, about 61 psi, about 62 psi, about 63 psi, about 64 psi, about 65 psi, about 66 psi, about 67 psi, about 68 psi, about 69 psi, about 70 psi, about 71 psi, about 72 psi, about 73 psi, about 74 psi, about 75 psi, about 76 psi, about 77 psi, about 78 psi, about 79 psi, about 80 psi, about 81 psi, about 82 psi, about 83 psi, about 84 psi, about 85 psi, about 86 psi, about 87 psi, about 88 psi, about 89 psi, about 90 psi, about 91 psi, about 92 psi, about 93 psi, about 94 psi, about 95 psi, about 96 psi, about 97 psi, about 98 psi, about 99 psi, about 100 psi, about 101 psi, about 102 psi, about 103 psi, about 104 psi, about 105 psi, about 106 psi, about 107 psi, about 108 psi, about 109 psi, about 110 psi, about 111 psi, about 112 psi, about 113 psi, about 114 psi, about 115 psi, about 116 psi, about 117 psi, about 118 psi, about 119 psi, about 120 psi, about 121 psi, about 122 psi, about 123 psi, about 124 psi, about 125 psi, about 126 psi, about 127 psi, about 128 psi, about 129 psi, about 130 psi, about 131 psi, about 132 psi, about 133 psi, about 134 psi, about 135 psi, about 136 psi, about 137 psi, about 138 psi, about 139 psi, about 140 psi, about 141 psi, about 142 psi, about 143 psi, about 144 psi, about 145 psi, about 146 psi, about 147 psi, about 148 psi, about 149 psi, about 150 psi, about 151 psi, about 152 psi, about 153 psi, about 154 psi, about 155 psi, about 156 psi, about 157 psi, about 158 psi, about 159 psi, about 160 psi, about 161 psi, about 162 psi, about 163 psi, about 164 psi, about 165 psi, about 166 psi, about 167 psi, about 168 psi, about 169 psi, about 170 psi, about 171 psi, about 172 psi, about 173 psi, about 174 psi, about 175 psi, about 176 psi, about 177 psi, about 178 psi, about 179 psi, about 180 psi, about 181 psi, about 182 psi, about 183 psi, about 184 psi, about 185 psi, about 186 psi, about 187 psi, about 188 psi, about 189 psi, about 190 psi, about 191 psi, about 192 psi, about 193 psi, about 194 psi, about 195 psi, about 196 psi, about 197 psi, about 198 psi, about 199 psi, or about 200 psi from a prefilled syringe using a HVAI.

[0184] In one embodiment, the starting pressure refers to the starting pressure exerted on the subject’s tissue upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the starting pressure refers to the starting pressure exerted the syringe stopper of the prefilled syringe in a HVAI as the disclosed formulation is delivered to a subject. In one embodiment, the starting pressure refers to the starting pressure exerted on the formulation upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the starting pressurerefers to the starting pressure exerted on the prefilled syringe upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the starting pressure refers to the starting pressure generated by the HVAI device upon administration of the formulation from a prefilled syringe using a HVAI.

[0185] In one embodiment about 3 mb to about 50 mb of the formulation disclosed herein is administered to the subject with an ending pressure of about 1 psi to about 250 psi, about 1 psi to about 225 psi, about 1 psi to about 200 psi, about 10 psi to about 200 psi, about 10 psi to about 175 psi, about 10 psi to about 150 psi, about 10 psi to about 125 psi, about 15 psi to about 125 psi, about 15 psi to about 100 psi, about 15 psi to about 80 psi, about 15 psi to about 75 psi, or about 20 psi to about 75 psi from a prefilled syringe using a HVAI. In one embodiment about 3 mb to about 50 m of the formulation disclosed herein is administered to the subject with an ending pressure of about 20 psi, about 25 psi, about 30 psi, about 35 psi, about 40 psi, about 45 psi, about 50 psi, about 55 psi, about 60 psi, about 65 psi, about 70 psi, or about 75 psi from a prefilled syringe using a HVAI. In one embodiment about 3 mL to about 50 mL of the formulation disclosed herein is administered to the subject with an ending pressure of about 20 psi, about 21 psi, about 22 psi, about 23 psi, about 24 psi, about 25 psi, about 26 psi, about 27 psi, about 28 psi, about 29 psi, about 30 psi, about 31 psi, about 32 psi, about 33 psi, about 34 psi, about 35 psi, about 36 psi, about 37 psi, about 38 psi, about 39 psi, about 40 psi, about 41 psi, about 42 psi, about 43 psi, about 44 psi, about 45 psi, about 46 psi, about 47 psi, about 48 psi, about 49 psi, about 50 psi, about 51 psi, about 52 psi, about 53 psi, about 54 psi, about 55 psi, about 56 psi, about 57 psi, about 58 psi, about 59 psi, about 60 psi, about 61 psi, about 62 psi, about 63 psi, about 64 psi, about 65 psi, about 66 psi, about 67 psi, about 68 psi, about 69 psi, about 70 psi, about 71 psi, about 72 psi, about 73 psi, about 74 psi, or about 75 psi from a prefilled syringe using a HVAI.

[0186] In one embodiment, the ending pressure refers to the ending pressure exerted on the subject’s tissue upon administration of the disclosed formulation from a prefilled syringe using a HVAI. In one embodiment, the ending pressure refers to the ending pressure exerted the syringe stopper of the prefilled syringe in a HVAI as the disclosed formulation is delivered to a subject. In one embodiment, the ending pressure refers to the ending pressure exerted on the formulation upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the ending pressure refers to the ending pressure exerted on the prefilled syringe upon administration of the formulation from a prefilled syringe using a HVAI. In one embodiment, the ending pressure refers to the ending pressure generated by the HVAI device upon administration of the formulation from a prefilled syringe using a HVAI.

[0187] In one embodiment, the administration of the formulation to the subject requires less applied force when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

[0188] In one embodiment, about 3 mL to about 50 mb of the formulation disclosed herein is administered at a rate of about 0.05 mL / sec to about 1.0 mL / sec with an applied force of about 10 N to about 200 N about 20 N to about 150 N, about 10 N, about 20 N, about 30 N, about 40 N, about 50 N, about 60 N, about 70 N, about 80 N, about 90 N, about 100 N, about 110 N, about 120 N, about 130 N, about 140 N, about 150 N, about 160 N, about 170 N, about 180 N, about 190 N, or about 200 N.

[0189] In one embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.14 mL / sec to about 0.21 mL / sec with an applied force of about 10 N to about 45 N. In one embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.14 mL / sec to about 0.21 mL / sec using a 25 gauge needle with an applied force of about 10 N to about 45N. In one embodiment, about 5 mL of the formulation is administered to the subject at a rate of aboutO.14 mL / sec to about 0.21 mL / sec using a 25 gauge needle with an applied force of about 15 N to about 25 N. In another embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.14 mL / sec to about 0.21 mL / sec using a 25 gauge needle with an applied force of about 22 N to about 40 N. In one embodiment, the about 10 mL formulation is administered to the subject at a rate of about 0.32 mL / sec to about 0.42 mL / sec with an applied force of about 25 N to about 50 N.

[0190] In one embodiment, the applied force at which the formulation is administered to the subject at a specific rate is dependent upon the gauge of the needle used to deliver the formulation to the subject. In one embodiment, the applied force at which the formulation is administered to the subject at a specific rate is dependent upon the gauge of the needle used to deliver the formulation to the subject and the inner diameter of the needle used to deliver the formulation to the subject. Therefore, in one embodiment, the use of a 25 gauge needle with a larger inner diameter (i.e., a thin wall needle such as a Terumo needle) will require less applied force to administer the disclosed formulation to a subject at a specific rate than the use of a 25 gauge needle with a smaller inner diameter (such as a BD needle). In one embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.14 mL / sec to about 0.21 mL / sec using a 25 gauge needle with an applied force of about 15 N to about 25 N, wherein the needle is a thin wall needle (e.g., a Terumo needle). In another embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.14 mL / sec to about 0.21 mL / sec using a 25 gauge needle with an applied force of about 22 N to about 40 N, wherein the needle is not a thin wall needle (e g., a BD needle).

[0191] In one embodiment, the about 10 mb formulation is administered to the subject using a 25 gauge needle at a rate of about 0.32 mL / sec to about 0.42 mL / sec with an applied force of about 25 N to about 50 N.

[0192] In one embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.14 mL / sec to about 0.21 mL / sec with an applied force of about 10 N, about 11 N, about 12 N, about 13 N, about 14 N, about 15 N, about 16 N, about 17 N, about 18 N, about 19 N, about 20, about 21 N, about 22 N, about 23 N, about 24 N, about 25 N, about 26 N, about 27 N, about 28 N, about 29 N, about 30 N, about 31 N, about 32 N, about 33 N, about 35 N, about 36 N, about 37 N, about 38 N, about 39 N, about 40 N, about 41 N, about 42 N, about 43 N, about 44 N, or about 45 N. In one embodiment, about 5 mL of the formulation is administered to the subject at a rate of about 0.14 mL / sec to about 0.21 mL / sec using a 25 gauge needle with an applied force of about 10 N, about 11 N, about 12 N, about 13 N, about 14 N, about 15 N, about 16 N, about 17 N, about 18 N, about 19 N, about 20, about 21 N, about 22 N, about 23 N, about 24 N, about 25 N, about 26 N, about 27 N, about 28 N, about 29 N, about 30 N, about 31 N, about 32 N, about 33 N, about 35 N, about 36 N, about 37 N, about 38 N, about 39 N, about 40 N, about 41 N, about 42 N, about 43 N, about 44 N, or about 45 N.

[0193] In one embodiment, the about 10 mL formulation is administered to the subject at a rate of about 0.32 mL / sec to about 0.42 mL / sec with an applied force of about 25 N, about 26 N, about 27 N, about 28 N, about 29 N, about 30 N, about 31 N, about 32 N, about 33 N, about 35 N, about 36 N, about 37 N, about 38 N, about 39 N, about 40 N, about 41 N, about 42 N, about 43 N, about 44 N, about 46 N, about 47 N, about 48 N, about 49 N, or about 50 N. In one embodiment, about 10 mL formulation is administered to the subject at a rate of about 0.32 mL / sec to about 0.42 mL / sec using a25 gauge needle with an applied force of about 25 N, about 26 N, about 27 N, about 28 N, about 29 N, about 30 N, about 31 N, about 32 N, about 33 N, about 35 N, about 36 N, about 37 N, about 38 N, about 39 N, about 40 N, about 41 N, about 42 N, about 43 N, about 44 N, about 45 N, about 46 N, about 47 N, about 48 N, about 49 N, or about 50 N.

[0194] The HVAI may deliver the full deliverable volume of the formulation in 5 seconds, 10 seconds, 15 seconds, 20 seconds, 25 seconds, 30 seconds, 35 seconds, 40 seconds, 45 seconds, 50 seconds, 55 seconds, 60 seconds, 70 seconds, 80 seconds, 90 seconds, 100 seconds, 110 seconds, or 120 seconds.

[0195] In one embodiment, the hyaluronidase enzyme allows the formulation to be administered to the subject faster than a comparable formulation that does not comprise the hyaluronidase enzyme. In one embodiment, the disclosed formulation can be administered about 20%, about 22%, about 24%, about 26%, about 28%, about 30%, about 32%, about 34%, about 36%, about 38%, about 40%, about42%, about 44%, about 46%, or about 48% faster than a comparable formulation that does not comprise the hyaluronidase enzyme when both are administered using the same HVAI fitted with a 23 gauge needle. Tn one embodiment, the disclosed formulation can be administered about 2%, about 4%, about 6%, about 8%, about 10%, about 12%, about 14%, about 16%, about 18%, or about 20% faster than a comparable formulation that does not comprise the hyaluronidase enzyme when both are administered using the same HVAI fitted with a 25 gauge needle.

[0196] In one embodiment, the injection of a high volume of the disclosed formulation in a subject leads to fewer side effects in the subject compared to an identical subject administered the same volume of a comparable formulation that does not comprise the hyaluronidase enzyme. In one embodiment, the injection of a high volume disclosed elsewhere herein with the disclosed formulation has reduced back leakage compared similar formulation that does not comprise the hyaluronidase enzyme. In one embodiment, the back leakage is reduced about 54%, about 56%, about 58%, about 60%, about 62%, about 64%, about 66%, about 68%, about 70%, about 72%, about 74%, about 76%, or about 78% when a high volume of the disclosed formulation is administered to a subject using a HVAI fitted with a 23 gauge needle compared to a similar formulation that does not comprise the hyaluronidase enzyme. In one embodiment, the back leakage is reduced about 62%, about 64%, about 68%, about 70%, about 72%, about 74%, about 76%, about 78%, about 80%, about 82%, about 84%, or about 86% when a high volume of the disclosed formulation is administered to a subject using a HVAI fitted with a 25 gauge needle compared to a similar formulation that does not comprise the hyaluronidase enzyme.

[0197] In one embodiment, the swelling (bleb) volume is reduced following the injection of the disclosed formulation into a subject when compared to a similar formulation that does not comprise the hyaluronidase enzyme. In various embodiments the swelling (bleb) volume is reduced by at least about 3%, or at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about 50%. In some embodiments the swelling (bleb) volume is reduced by between about 2% to about 60%, or about 2% to about 50%, or about 2% to about 30%, or about 2% to about 25%, or about 2% to about 20%, or about 2% to about 15%, or about 2% to about 10%.

[0198] In one embodiment, the swelling height is reduced following the injection of the disclosed formulation when compared to a similar formulation that does not comprise the hyaluronidase enzyme. In various embodiments the swelling height is reduced by at least about 3%, or at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about50%. In some embodiments the swelling height is reduced by between about 5% to about 60%, or about 5% to about 50%, or about 5% to about 50%, or about 5% to about 25%, or about 5% to about 20%, or about 5% to about 15%, or about 5% to about 10%.

[0199] In one embodiment, the swelling size is reduced following the injection of the disclosed formulation when compared to a similar formulation that does not comprise the hyaluronidase enzyme. In various embodiments the swelling size is reduced by at least about 3%, or at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about 50%, or at least about 55%, or at least about 60%, or at least about 65%, or at least about 70%. In some embodiments the swelling size is reduced by between about 5% to about 80%, about 5% to about 75%, 5% to about 70%, about 5% to about 65%, 5% to about 60%, or about 5% to about 50%, or about 5% to about 50%, or about 5% to about 25%, or about 5% to about 20%, or about 5% to about 15%, or about 5% to about 10%.

[0200] In one embodiment, the swelling area is reduced following the injection of the disclosed formulation when compared to a similar formulation that does not comprise the hyaluronidase enzyme. In various embodiments the swelling area is reduced by at least about 2%, or at least about 3%, or at least about 4%, or at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about 50%. In some embodiments the swelling area is reduced by between about 2% to about 60%, or about 2% to about 50%, or about 2% to about 30%, or about 2% to about 25%, or about 2% to about 20%, or about 2% to about 15%, or about 2% to about 10%.

[0201] In one embodiment, the swelling induration following the initial injection of the disclosed formulation is minimized compared to a similar formulation that does not comprise the hyaluronidase enzyme. In various embodiments the swelling induration is reduced by at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 25%, or at least about 30%, or at least about 35%, or at least about 40%, or at least about 45%, or at least about 50%, or at least about 55%, or at least about 60%, or at least about 65%. or at least about 70%. In some embodiments the swelling induration is reduced by between about 20% to about 80%, or about 20% to about 70%, or about 20% to about 60%, or about 20% to about 50%, or about 20% to about 40%, or about 20% to about 70%, or about 20% to about 60%.

[0202] In one embodiment, the swelling resolves quicker when the disclosed formulation is injected compared to a similar formulation that does not comprise the hyaluronidase enzyme.

[0203] In one embodiment, the disclosed formulation permits for more consistent delivery (i.e., time to delivery, reduction in bleb swelling volume, height and induration) from injection to injection, compared to a similar formulation that does not comprise the hyaluronidase enzyme.

[0204] In one embodiment, the disclosed formulation permits for faster delivery of the full volume from a HVAI than a comparable formulation that does not comprise the hyaluronidase enzyme which results in less pain and discomfort for the subject. In various embodiments, the delivery time is reduced by at least about 3%, or at least about 5%, or at least about 10%, or at least about 15%, or at least about 20%, or at least about 25%, or at least about 30%. In some embodiments the delivery time is reduced by between about 5% to about 30%, or about 5% to about 25%, or about 5% to about 20%, or about 5% to about 15%, or about 5% to about 10%.Example 1: Assessment of a Ten mL Subcutaneous Angled Injection Using an HVAI and 25gNeedleSUMMARY

[0157] The objective of this study was to assess the impact of an angled injection platform on the subcutaneous (SC) delivery of a representative biotherapeutic both with and without recombinant human hyaluronidase PH20 (rHuPH20). Yucatan miniature pigs were used in this study due to their high degree of similarity with human skin architecture. Each animal received two 10 mL SC injections on the lower abdominal region of the animal. An immunoglobulin solution prepared at 120 mg / mL (Ig- 120) was used either alone or co-mixed with rHuPH20 at a concentration of 2000 U / mL. The test solution was delivered using a high-volume auto-injector (HVAI). To accommodate an angled delivery a custom platform was created that held the HVAI at a 45° angle relative to the surface of the animal and gave an exposed needle depth of approximately 7.5 mm.

[0158] The addition of rHuPH20 to the Ig-120 solution resulted in a reduction in the injection time for the HVAI (-15%) with much less variability for injections of Ig-120 + rHuPH20 compared to Ig- 120 alone. Back-leakage was low and approximately equivalent between both treatment groups. Measurement of bleb size showed a significant reduction in volume and area immediately postinjection for injections of Ig-120 + rHuPH20 compared to Ig-120 with faster bleb resolution (swelling size and induration) for injections that contained rHuPH20. Because the HVAI has been used in earlier studies using a vertical insertion angle, direct comparison of the two studies showed that injection times were similar to that measured in a study that utilized the HVAI and similar Ig solutions and (Ig- 120: 50.8 vs. 47.8; Ig-120 + rHuPH20: 43.3 vs. 43.4 sec). Back-leakage was greater for vertical injections of Ig-120 compared to this study (16.2 mg vs. 63.1 mg) which may be a result of the angled delivery of the test solution. Bleb height was lower in this study compared to vertical injections (Ig-120: 7.3 mm vs. 8.4 mm; Ig-120 + rHuPH20: 4.4 mm vs. 5.3 mm) with a greater reduction in volume (57% vs. 44%) seen for the angled injections. Qualitative assessments of the post-injection swelling showed a reduction in swelling size for the angled injections compared to the vertical injections (Ig- 120: 3.3 vs. 3.4; Ig-120 + rHuPH20: 2.7 vs. 2.9). Notably, the induration assessments post-injection were reduced for angled injections compared to vertical injections (Ig-120: 3.0 vs. 3.4; Ig-120 + rHuPH20: 1.9 vs. 2.9).INTRODUCTION

[0159] Current auto-injectors (AIs) are limited to extremely small volumes (typically < 2.25 mb), limiting their usefulness for delivery of larger volumes. But current biologies typically may have dose volumes of 5-50 mb. For larger volumes higher flow rates are required to make use of an Al practical. Currently 30 seconds is a recommended amount of time that a device can be held in place during selfadministration to prevent fatigue and potential interruption of the injection. A prototype high volume auto-injector (HVAI) was created to deliver larger volumes (10 mb) of a biotherapeutic in this timeframe. This prototype device has been shown both pre-clinically and clinically to facilitate the delivery of high-volume biotherapeutics in this approximate timeframe.

[0160] rHuPH20 has been shown to facilitate the SC administration of fluids and drugs by transiently and locally depolymerizing hyaluronan (HA) in the extracellular matrix (ECM). The depolymerization of HA reduces tissue backpressure in the SC space that subsequently allows for rapid, large volume administration of drugs. Understanding the impact of rHuPH20 on HVAI performance is important for future HVAI designs.

[0161] Angled delivery of a biotherapeutic using an HVAI may have differences in performance compared to vertical needle insertion (e.g., flow rates, bleb size and dispersion, etc.), as flow from the injection site is directed away from the injection needle rather than being forced down towards the muscle layer. However, the impact of this change in route of delivery is not well characterized. Therefore, the objective of this study was to determine the impact of angled needle insertion and delivery on the performance of a clinically tested HVAI using the mini-pig as an animal model. The mini-pig model has been selected due to the high degree of similarity of the subcutaneous space to that of humans. Previous studies using a mini-pig model have demonstrated the translatability of the model for use in pre-clinical (Kang et al., 2013) and auto-injector studies (Shi et al., 2021).EXPERIMENTAL DESIGN AND METHODSTest and Control Articles

[0162] Ig-120 Test SolutionDescription: Clear liquid reconstituted from lyophilized powderConcentration: 120 mg / mLFormulation: 20 mM Histidine, 130 mM sodium chloride, pH 6.5Lot number: NB 1090-49Storage Conditions: <-65 °CHandling Conditions: Standard laboratory precautionsSupplier: Biomed SupplyFormulated by: Halozyme Product Development

[0163] Recombinant Human Hyaluronidase PH20 (rHuPH20)Description: Clear colorless liquidLot number: l-FIN-3928Concentration: 9.3 mg / mLEnzyme activity: 1,039,763 U / mLFormulation: 10 mM Histidine, 130 mM sodium chloride, pH 6.5Storage Conditions: < 70°CHandling Conditions: Standard laboratory precautions Supplier: Halozyme Therapeutics, Inc.

[0164] Ig Dilution BufferDescription: Clear colorless liquidFormulation: 20 mM histidine, 130 mM sodium chloride, pH 6.5Batch / Lot: NB 1078- 18Storage Conditions: 2-8°CHandling Conditions: Standard laboratory precautions Supplier: Halozyme Therapeutics, IncFormulation

[0165] Preparation of Test Solutions. Two test solutions were utilized for this study. The first test solution was Ig-120 alone. The second test solution was Ig-120 + rHuPH20. To ensure sufficient test solution for syringe filling 75 mL of each test solution was prepared.

[0166] To prepare the co-mix of Ig-120 + rHuPH20, 0.144 mL of rHuPH20 was added to the 75 mL of Ig-120 to yield a test solution of Ig-120 + rHuPH20 with a final concentration of 2000 U / mL. After preparation the test solutions were used for syringe filling.

[0167] Device Syringe Filling. Filling of all device syringes occurred in a sterile fill and finish hood under aseptic conditions. Cyclic olefin copolymer (COC) syringes were filled with 10.1 mL of either Ig-120 alone or the co-mix of Ig-120 + rHuPH20. To fill the syringes a sterile rubber plungerwas initially inserted into the sterile syringe to a loading position. A syringe pump was then used to fill the syringe with 10.1 mL of the test solution. After adding the test solution to the syringe, the rubber plunger was brought to the final fill position (to remove residual air in the syringe). The syringe was then be capped and stored at 2-8°C until used for device assembly.

[0168] Device Assembly. Assembly of all devices occurred in a sterile fill and finish hood under aseptic conditions. The syringe cap was removed and replaced using a 25Gx 1-inch Becton Dickinson (BD) needle with the needle remaining capped. The needle and syringe were placed in a proprietary assembly jig and the HVAI devices constructed by trained Halozyme engineering staff. Final assembly included the placement of a needle guard that brought the exposed needle length to 10 mm, which gave an approximate vertical injection depth of -1.5 mm in the SC space when the device was mounted into the angled platform.Animal Description

[0169] Species: Pig (Sus scrofa domestica)Strain: Yucatan MiniatureSex: FemaleAge: ~ 3-4+ months Weight: 14-16 kg Quantity: 6 animals Source: Premier BioSource (Ramona, CA)Table 1 : Summary of Animal Weights (Day of Treatment)

[0170] Husbandry. Animals were housed in steel pens with automatic water provided ad libitum. Animals were fed twice daily (AM and PM) but were kept NPO after midnight on the day of the study. The room environment was set to maintain a temperature of ~19-23°C and a relative humidity of 40-70%, with a 12 hour light / 12 hour dark time cycle. Animals were acclimated to the vivarium for a minimum of 3 days prior to study start.Test Materials

[0171] The test materials used in this Example are summarized in Table 2.Table 2: Summary of Test MaterialsEXPERIMENTAL DESIGN

[0172] In this study, two 10 mL injections were administered to the abdomen of a Yucatan miniature pig. On one side of the abdomen a test solution of Ig-120 alone was administered and a second injection of Ig-120 + rHuPH20 on the contralateral side of the animal. All test solutions containing rHuPH20 were formulated at 2000 U / mL. The treatments for each animal are summarized in Table 3.Table 3: Description of Treatments

[0173] The injections were administered using a HVAI device that was loaded into a custom 3D printed platform that allows the needle to be inserted to skin at a 45° angle. The platform was designed to make the final needle depth of 7.5 mm (measured vertically from the skin surface). The HVAI was oriented so that the needle bevel faced upwards when mounted in the platform.

[0174] The length of each injection was recorded using a stopwatch. Immediately post-injection the platform with HVAI was removed and back-leakage collected for 30 seconds using a pre-weighed eye spear. Swelling size was then measured using a digital caliper and by obtaining a 3D image capture immediately post-injection (TO). Caliper measurement of bleb size was also taken and at 15 minutes (T15) and 30 minutes (T30) post-injection. Standard photographs were obtained at TO, T15, T30, T2h (2h post-injection) and T24h (24h post-injection).

[0175] Additional post-injection qualitative injection site evaluations for erythema, swelling and induration were performed at TO, T15, T30 and T2h with a final evaluation at approximately T24h post-injection. In summary the endpoints for the study will be: (1) measurement of HVAI injection time (digital stopwatch); (2) collection and measurement of back-leakage weight using an eye spear; (3) caliper measurement of bleb size post-injection using digital caliper at TO, T15 and T30; (4) measurement of bleb volume, area and height at TO using 3D imaging; (5) assessment of blebs for erythema, swelling size and induration at times TO, T15, T30, T2h and T24h; (6) photographic documentation of all injection sites: pre-inj ection, TO, T15, T30, T2h and T24h.PROCEDURES

[0176] On the day prior to the study HVAI devices (one Ig-120; one Ig-120 + rHuPH20) were withdrawn from refrigerated storage and brought to room temperature for at least 30 minutes prior to use. The first injection for each animal was always Ig-120 followed by a second injection on the contralateral side of the animal with Ig-120 + rHuPH20. Injection sites were located on the left and right abdominal regions, ~5 cm cranially from the inguinal fold towards the midline and ~3 cm towards the midline of the animal.

[0177] Prior to the start of study, animals were assessed for general health, and body weights were collected. Animals were anesthetized with isoflurane gas and placed in dorsal recumbence on a heated surgical table and were maintained under isoflurane gas for the entire duration of the procedure. The abdominal region was cleaned with Nolvasan and the injection site wiped with gauze containing 70% isopropanol and wiped dry with sterile gauze. Each of the injection sites were marked with a permanent marker and then photographed with the standard and 3D cameras prior to HVAI needle insertion. The HVAI was mounted into the platform and the needle bevel confirmed to face upward. The HVAI was unlocked and the needle cap was removed and discarded prior to needle insertion.

[0178] The skin was pinched at the injection site and the HVAI / platform with exposed needle was inserted into the pinched skin until the platform was flush with the skin. The pinched skin was then allowed to relax and lie flush with the skin surface. After needle insertion the injection started by pushing the activation button. The injection was timed using a digital stopwatch. Upon completion ofinjection the injection time was recorded and the platform with HVAI withdrawn from the skin. Test solution back-leakage at the injection site was absorbed to a pre-weighed eye spear for 30 seconds. The weight of the eye spear was measured using an analytical balance with an accuracy of 0.1 mg and recorded. The margins of the injection site bleb were marked with a permanent marker and measured for length, width, and height using a digital caliper and recorded then photographed with the standard and 3D cameras immediately post-injection (TO). Caliper measurements were also taken at T15 and T30 post-injection. After the second injection two additional HVAIs (one with Ig-120 and one with Ig-120 + rHuPH20) were removed from refrigerated storage and brought to room temperature for use with the following animal. Local injection sites were also qualitatively assessed and graded for erythema severity, swelling size appearance, and firmness immediately after injection, using a scoring system described in Table 4 , Table 5 and Table 6 respectively. Any additional observations were noted at this time. The injection site was then qualitatively assessed at TO, T15 and T30 and photographed using a standard camera while the animal was still under anesthesia. The injection sites were again qualitatively assessed and photographed at T2h but the animal was conscious and handheld by an animal technician.

[0179] After the first injection and while the animal was still sedated, the identical procedure was repeated on the contralateral side of the animal with the second test solution (Ig-120 + rHuPH20). Thirty minutes after completion of the second injection (and after the final assessments at T30 minutes), the animal was removed from anesthesia and returned to its home cage for recovery. On the following day (at approximately 24 hours) the animal was re-anesthetized, and both of the injection sites assessed for erythema, induration and swelling, and photographed. Following the ~T24h assessment, the animal was humanely euthanized using an injectable euthanasia drug.Table 4: Grading Scale for Erythema FormationTable 5: Grading Scale for Swelling Size FormationTable 6: Grading Scale for Swelling Firmness (Induration)CALCULATIONS AND STATISTICAL METHODS

[0180] Assessment of Injection Time. The time required for each injection was measured using a digital stopwatch and recorded after each injection.

[0181] Assessment of Swelling Volume, Area and Height Using Caliper Measurement and 3D Imaging. Volume and area of post-injection swelling were measured using both caliper measurement and 3D camera image analysis. For caliper measurements a digital caliper was utilized to measure length, width and height of the bleb that formed post-injection. The length and width are defined as the edge to edge measurements of the bleb (i.e., diameter) along their longest axes. These values were manually recorded, and the volume determined using the formula for half of an ellipsoid Vol = (2 / 3)*7r*A*B*C where A = Length / 2, B = Width / 2 and C = Height.

[0182] 3D imaging was applied as a longitudinal methodology to measure post-injection swelling. By obtaining high definition pre- and post-injection 3D images the distances between two registered surfaces can be determined. The camera captures images using a factory calibrated bifocal imaging system to measure distance between surfaces. Surface registration was performed using a multipoint method that utilized common landmarks between the pre-inj ection image and the post-injection image. Using the proprietary software, the volume, area, and height of the post-injection swelling was calculated for each injection (Canfield Biosciences, Inc.).

[0183] Statistical comparisons for Ig-120 and Ig-120 + rHuPH20 were performed using an unpaired parametric t-test (Prism vl 0.1.1, GraphPad Software, Inc., San Diego, California). All hypotheses’ tests were performed at a 5% significance level, thus considered significant if p<0.05.RESULTS AND DISCUSSION

[0184] Pre-study Enzymatic Activity Testing. The co-mix of Ig-120 + rHuPH20 was prepared on the day prior to study start and the enzyme activity of the test solution was measured using an in vitro activity assay (VV-QUAL-00675; formerly TM010). The results of this activity testing are shown in Table 7.Table 7: Summary of Pre-Study Activity Testing

[0185] Measurement of Needle Depth Vertical Displacement. The exposed needle was placed in the skin at approximately a 45° angle. Therefore, the predicted vertical needle depth (a) can estimated using the formula: sin 45° = (a) / 10 mm = (I / A / 2)(a) = 7.1 mm

[0186] The length of each needle in the completed device was measured using a steel rule and is shown in Table 8. Both treatment groups had the same distribution of needle depths.Table 8: Measurement of Exposed Needle Lengths (Vertical Depth)Pre- and Post-injection Quantitative Measurements

[0187] Measurement of HV Al Injection Times. The time for each injection was measured using a stopwatch and is shown in Table 9 and Fig. 8. There was approximately a 15% decrease in delivery time with less variability when the Ig-120 was delivered co-mixed with rHuPH20 but this was not considered significant.Table 9: Measurement of HVAI Injection Times

[0188] Measurement of Post-injection Back-leakage . The back-leakage was collected using a preweighed eye spear for 30 seconds post-injection. The eye spear was then re- weighed, and the weight of the back-leakage recorded. The amount of back-leakage (mean ± SEM) for each test solution is shown in Table 10 with individual values shown in Fig. 9.Table 10: Summary of Post-injection Back-leakage

[0189] Assessment of Post-Injection Bleb Volume, Area and Height (Caliper Measurements) . The local injection site swelling (bleb) was marked and measured using a digital caliper. Bleb volume dispersion area and swelling height of each bleb was determined as described above and this data is summarized at the TO, T15 and T30 timepoints in Table 11, Table 12 and Table 13 and individual values shown in Fig. 10, Fig. 11, and Fig. 12. The post-injection swelling, area and height over time were also determined at T15 and T30 and the scores are shown in Fig. 13, Fig. 14, and Fig. 15, respectively.Table 11 : Bleb volume after injection of Ig-120 and Ig-120 + rHuPH20 over time using caliper measurement (mean ± SEM)Table 12: Bleb area after injection of Ig-120 and Ig-120 + rHuPH20 over time using caliper measurement (mean ± SEM)Table 13: Bleb height after injection of Ig-120 and Ig-120 + rHuPH20 over time using caliper measurement (mean ± SEM)

[0190] Assessment of Post-injection Bleb Shape, Volume, Area and Height (3D Imaging). Pre- and post-injection photographs were taken using a 3D imaging system (Canfield Scientific). This technology permits point-to-point alignment of these two images through multipoint surface registration. The distance between any two points is then represented using a colorimetric surface contour map. Regions where there is no difference between the two images are displayed in gray. Where the post-injection image is higher than the pre-injection image, the region is displayed in shades of blue. Where the post-injection image is lower than the pre-injection image the distance is displayed in shades of orange. The color intensity is proportional to the amount of distance measured between images with darker blue color indicating greater distance from the pre-injection image. Out of rangemeasurements (distances greater than 6 mm) are depicted in white. Bleb measurements of volume and height include regions out of range.

[0191] Each animal had a pre-injection 3D image taken of the injection site followed by a second image taken immediately post-injection and these images were mapped to each other using multipoint registration. These registered pre- / post-inj ection images were then used to calculate the bleb volume, height, circumference, length, and width for each bleb using proprietary software (Vectra Hl software; Canfield Sciences). Colorimetric surface contour maps of each post-injection bleb for Ig-120 and Ig- 120 + rHuPH20 are shown in Fig. 16. Mean bleb volume, area, and height are reported in Table 14 and Figs. 17, 18, and 20.Table 14: Bleb volume, area and height after injection of Ig-120 and Ig-120 + rHuPH20 assessed using 3D imaging (mean ± SEM)Qualitative Assessment of Local Injection Sites[00192J Qualitative assessments were taken immediately post-injection (TO), 15 minutes postinjection (T15), 30 minutes post-injection (T30), 2 hours post-injection (T2h) and at approximately 24 hours post-injection (T24). Only two evaluators were available for scoring at T24.

[0193] Post-injection Erythema. The post-injection erythema was evaluated at all timepoints (TO, T15, T30, T2h and T24h). Erythema was very mild and transient, having only been observed in 3 / 12 animals at TO and T15 with complete resolution by T30. No erythema was seen at T2h and T24h (data not shown). While the initial erythema scores for Ig-120 + rHuPH290 were slightly elevated compared to Ig-120, the mean score for this group was <1 (minimal). The erythema scores (Mean ± SEM) are shown in Table 15 and Fig. 20.Table 15: Qualitative assessment of post-injection erythema (T0-T15-T30-T2h)

[0194] Post-injection Swelling Size. Post-injection swelling size was significantly greater for Ig- 120 than for Ig-120 + rHuPH20. Scoring by three evaluators for swelling size (Mean ± SEM) are summarized in Table 16. Individual values immediately post-injection (TO) are shown in Fig. 21 and values scored over time (Mean ± SEM) are shown in Fig. 22.Table 16: Qualitative assessment of post-injection swelling size (T0-T15-T30-T2h)

[0195] Post-injection Induration. The induration (firmness) of the post-injection blebs for Ig-120 was found to be significantly greater than the blebs that resulted from injection of Ig-120 + rHuPH20 (p<0.05). Scoring by three evaluators for swelling size (Mean ± SEM) are summarized in Table 17.Individual values immediately post-injection (TO) are shown in Fig. 23 and values scored over time (Mean ± SEM) are shown in Fig. 24.Table 17: Qualitative assessment of post-injection induration (T0-T15-T30-T2h)PHOTOGRAPHS OF INJECTION SITES PRE- AND POST-INJECTION

[0196] The injection sites were photographed before and immediately post-injection and then at T15, T30, T2h and T24h. Photographic images are shown in Figs. 25-36.PROTOCOL DEVIATIONS AND IMPACT

[0197] The ASA setting was incorrect for the Pre-inj ection photo for AID #5189L resulting in an overexposed image. The pre-injection photo from the 3D imaging was substituted in Figure 18. There was no impact on the results of the study.

[0198] The photo taken for AID #5312L (Ig-120 + rHuPH20) at the T15 timepoint incorrectly had a time on the ruler of T30. The photo was correctly taken at the T15 timepoint but incorrectly labeled in the photo. There was no impact on the results of the study.

[0199] No pre-injection photo for AID #5179R was taken and no 3D analysis could be performed. An N=5 is indicated for this group.SUMMARY AND CONCLUSIONS

[0200] This study explored the delivery of an Ig solution with and without rHuPH20 using an angled platform so that the needle was inserted at approximately a 45° angle. The Ig solution was formulated at 120 mg / mL and the concentration of rHuPH20 used for the injections was 2000 U / mL. All test solutions were delivered using a HVAI.

[0201] Key observations include: (1) the mean injection time for Ig-120 + rHuPH20 was -15% faster than the mean injection time for Ig-120; (2) back-leakage was minimal for both test solutions; (3) post-injection bleb volume and area were found to be significantly less for injections of Ig-120 ± rHuPH20 compared to Ig-120 alone immediately post-injection (TO).

[0202] Qualitative scoring of post-injection swelling size showed an immediate reduction in bleb size for injections of Ig-120 + rHuPH20 compared to Ig-120 alone with much more rapid resolution over the first two hours post-injection. Swelling scores for Ig-120 + rHuPH20 were <1 by 2 hours post-injection whereas control injections of Ig-120 were still prominent at this timepoint.

[0203] Qualitative scoring of post-injection induration indicated that the firmness scores immediately post-injection for Ig-120 + rHuPH20 were significantly less than that for injections of Ig-120 alone. In addition, induration for injections of Ig-120 + rHuPH20 were scored <1 (minimal) by the T30 timepoint.

[0204] The mean injection time for Ig-120 + rHuPH20 using the angled platform (43.3 sec) was comparable to the mean injection time measured previously for a vertical needle placement (43.4 sec) with much less variability than control injections.

[0205] The results from this study were consistent with previous studies showing the contribution of rHuPH20 for reducing injection time, post-injection swelling and induration. However, these results were not materially different from an earlier study that showed similar bleb sizes, induration and resolution. Overall, these results suggested that the delivery using an angled platform does not appear to make a material contribution to the SC delivery of Ig-120.Example 2: Assessment of a Ten mL Subcutaneous Angled Injection Using a 25G Needle forDevelopment of an Auto-InjectorSUMMARY

[0206] This study examined the delivery of an Ig solution formulated at 120 mg / mL using a mock auto-injector. The test solution was delivered with and without rHuPH20 at a concentration of 2,000 U / mL. All injections were performed using a platform that allowed for angled needle insertion at a 45° angle. The test solution volume was 10 mL and was delivered in 30 seconds using a 20cc syringe and 25G needle with an insertion depth of 7.5 mm. The applied force to the syringe barrel was measured throughout the injection by attaching a load cell to the end of the syringe flange. In addition, the back-leakage was collected post-injection and quantified by weight. The post-injection swelling was measured using calipers and 3D imaging. After the injection three independent scorers evaluated the injection site for erythema, swelling size and induration over time (at times T=0, 15, 30 min, 2h and 24h) to assess the time for the resolution of the post-injection swelling.INTRODUCTION

[0207] The objective of this study was to determine if rHuPH20 may potentiate the development of a high volume auto-injection (Al) that is able to deliver clinically relevant volumes to the SC space at high flow rates using the mini-pig as an animal model. In particular, this study assessed the deliveryan Ig solution formulated at 120 mg / mL through a 25G needle when inserted into the SC space at a 45° angle using a platform and hand-held mock auto-injector device.TEST ARTICLES AND METHODSTest Articles

[0208] Human Gamma Globulin (Ig-120: 12% solution)Lot number: 1032-17 (Appendix A)Description: Lyophilized powder reconstituted at 120 mg / mLDate of Manufacture : 21 SEP2020Formulation: 10 mM Histidine, 130 mM Sodium Chloride, pH 6.5Storage Conditions: 2-8°CSupplier: BioMed SupplyFormulated by: Halozyme Product Development

[0209] Recombinant Human Hyaluronidase rHuPH20 (ENHANZE™ Drug Product)Lot number: 462-022 (Appendix B)Description: Clear and colorless solutionConcentration: 10 mg / mLDate of Manufacture: December 30, 2014Retest Date: February 2023Enzyme activity: 1,229,456 U / mLStorage: < 70°CFormulation: 10 mM Histidine, 130 mM sodium chloride, pH 6.5Handling Conditions: Standard laboratory precautionsSupplier: Halozyme Therapeutics, IncFormulation

[0210] Preparation of Test Solutions. The two test solutions administered in this study were Ig- 120 alone and Ig-120 + rHuPH20. These were prepared by addition of rHuPH20 from a concentrated stock to an Ig solution previously prepared at 120 mg / mL. The final concentration of rHuPH20 in the test solution was 2,000 U / mL.

[0211] Ig-120 was thawed at 2-8 °C overnight. The following day test solutions were prepared by adding rHuPH20 to the Ig-120 solution at room temperature. A concentrated stock of rHuPH20 was used for test article preparation (10 mg / mL; 1,229,456 U / mL). To prepare Ig-120 + rHuPH20, 488 pL of rHuPH20 was added to 300 mL of Ig-120. 75 mL of each test solution was then aliquoted intoindividual 100 mL glass vials and stored at 4°C until used for syringe filling on the day prior to the study.

[0212] The Ig-120 + rHuPH20 solution was tested for rHuPH20 activity prior to the start of the study using a micro-turbidity assay. The activity of the Ig-120 + rHuPH20 test solution was within 10% of target concentration and were deemed to be within acceptable range for use in the study. The test solution was prepared and stored at 2-8°C and tested for enzyme activity prior to study start.

[0213] At the end of the study dose retain samples that were obtained during the study procedure were tested for rHuPH20 activity. The activity of the Ig-120 + rHuPH20 test solutions were deemed to be within acceptable range. These values are summarized in Table 18 and Table 19.Table 18: Pre -Study Activity Testing of rHuPH20 Activity in Test SolutionsTable 19: Post-Study Activity Testing of rHuPH20 Activity in Test SolutionsANIMAL DESCRIPTION

[0214] Species: Pig (Sus scrofa domestica)Strain: Yucatan miniatureSex: FemaleAge: >3 monthsBody weight: 12-16 kg upon receiptQuantity: 6Source: Premier BioSource (Ramona, CA)

[0215] Husbandry. Animals were received on 9-September-2022 by the facility and allowed to acclimate prior to study start. Animals were group housed in steel pens with automatic water provided ad libitum. Animals were fed twice daily (AM and PM), except on study day (PM only). Room environment was set to maintain a temperature of ~17-27°C and a relative humidity of 40-70%, with a 12 hour light / 12 hour dark time cycle. Animals were allowed to acclimate to the facility for a minimum of 3 days prior to study onset.EXPERIMENTAL DESIGN

[0216] In this study, two 10 mL injections were administered to the abdomen of a Yucatan miniature pig. On one side of the abdomen a test solution of Ig-120 alone was administered. One the contralateral side of the animal a second test solution of Ig-120 + rHuPH20 was administered. All test solutions containing rHuPH20 were formulated at 2000 U / mL. The location of the injection sites was randomized with three injections of each test solution on the left side of the animal and three injections on the right side of an animal. The injections were performed using a mock auto-injector device mounted inside a 3D printed platform allowing for a 45° insertion angle. The needle was positioned in the handle so that the bevel of the needle was facing upward (toward the top of the SC space). The treatments for each animal are summarized in Table 20, Description of Treatments.Table 20: Description of Treatments

[0217] Quantitative endpoints included in this study were measurement of applied force to the syringe barrel during the injection, post-injection swelling (bleb) volume, area and height. In addition, the post-injection back-leakage of test article was collected from the injection site for 30 seconds after the removal of the needle using an eye-spear to absorb any leakage and quantified by weight. The volume of the injection site blebs was determined by digital caliper measurement (length, width, and height) as well as by 3D camera imaging. Additional post-injection qualitative injection site evaluations for erythema, swelling and induration was performed immediately post injection (TO) and at 15 minutes post-injection (T15), 30 minutes post-injection (T30), 2 hours post-injection (T2h) and at approximately 24 hours post-injection (T24h) post-injection. Qualitative assessments of the injection sites were performed while the animal was under anesthesia for the TO, T15, T30 and T24h timepoints while the T2h assessment was performed while the animal was conscious and hand-heldby an animal technician. Standard photographs were obtained both pre-inj ection and at times TO, T15, T30, T2h and T24h post-injection. After euthanasia a 12 mm punch biopsy was obtained from the injection site and fixed in 10% formalin. In summary the endpoints for the study were: (1) applied force during the injection; (2) measurement of back-leakage post-injection; (3) measurement of bleb size (length / width / height) post-injection (caliper); (4) measurement of bleb size (volume, height, area) using 3D imaging; (5) assessment of blebs for erythema, swelling size and induration at times TO, T15, T30, T2h and T24h; (6) measurement of injection site temperature (pre-inj ection and postinjection); (7) assessment of injection site post-injection (24h) by histology.STUDY PROCEDURE

[0218] Prior to start of study, animals were assessed for general health, and body weights were collected. On the day prior to the study test articles (~17 mL) were drawn into a 20 mL syringe, capped and stored at 2-8 °C. On the day of the study the syringes were removed from 2-8 °C and brought to room temperature for at least 30 minutes but no more than 4 hours. Dose retains taken during the study procedure were kept at room temperature until transferred back to Halozyme Product Development for enzymatic testing on the day following the study procedure.

[0219] Animals were anesthetized with isoflurane gas and placed in dorsal recumbence on a foam wedge placed on a heated surgical table and were maintained under isoflurane gas for the entire duration of the procedure. The abdominal region was cleaned with Nolvasan followed by wiping the injection site with gauze containing 70% isopropanol and wiped dry with sterile gauze. Injection sites were located on the left and right abdominal regions, ~6 cm cranially from the inguinal fold towards the midline and ~3 cm towards the midline of the animal. Each of the injection sites was marked with a permanent marker and then photographed with the standard and 3D cameras prior to needle insertion. The temperature of the skin at the injection site was recorded prior to the start of the injection using an infrared thermometer. The initial injection for each animal was the control solution (Ig-120 alone). The second injection on the contralateral side of the animal was the test solution containing rHuPH20 (Ig-120 + rHuPH20).

[0220] Assembly of mock device'. The mock device was prepared by attaching a capped 25G x 1 inch Leur-lok needle to the male end of a 21 -inch extension set. The extension set was then be routed through the inside of the mock device and the needle was firmly seated in place in the end of the device. The device with needle attached was then inserted into the platform. The length of the needle projecting from the end of the mock device was confirmed to be 10.6 mm ± 0.5 mm providing an injection depth of 7.5 mm (measured vertically). The needle remained capped until just prior to vertical needle insertion. The 20cc syringe that contains the test solution was uncapped, attached tothe female end of the extension set and then the hardware was primed to the needle tip with the test solution and then the syringe was placed into the syringe pump. The load cell was then attached to the end of the syringe plunger. After zeroing the load cell, the applied force readings were initiated. The pump block was positioned so that it abutted the end of the syringe plunger-load cell with minimal contact force and was then locked into place. The needle-device-platform was inserted at a 45° angle into the marked injection site and held in place by hand at the predetermined depth of ~7.5 mm. Once applied force readings were confirmed the syringe pump was started to begin injection of the test article at the designated flow rate of 20 mL / min. Upon completion of injection the needle and platform was removed, the pressure on the syringe pump block removed and the applied force data collection was stopped. Test solution back-leakage was then absorbed to a tared eye-spear for 30 seconds by blotting the injection site. The margins of the injection site bleb were marked with a permanent marker and measured for length, width, and height using a digital caliper and recorded then photographed with the standard and 3D cameras. The injection site was then qualitatively scored by three independent evaluators for appearance and severity of erythema, swelling / bleb size, and firmness (induration) using a 5-point scoring system (a modified Draize Test) based on the 1992 OECD guidelines for grading skin reactions. The evaluators were blinded to each other’s scores After the first injection, the procedure was repeated on the contra-lateral side of the animal using the other test solution (Ig-120 + rHuPH20). Qualitative scoring for erythema, swelling, and induration were collected by all 3 evaluators again at 15 min., 30 min, 2 hr, and approximately 24 hr post injections. Photographs with the standard camera were collected at each of these timepoints. Following the final assessment, the animal was humanely euthanized using a ready for use solution of sodium pentobarbital and sodium phenytoin (Euthasol®).CALCULATIONS AND STATISTICAL METHODS

[0221] Assessment of Applied Force: Applied force, as measured via a load cell attached to the end of syringe plungers, was recorded using SensorVUE software (Loadstar Sensors), at a rate of 2 Hz and the mean applied force over the entire injection period was calculated.

[0222] Assessment of Local Swelling Volume and Area Using Caliper Measurement and 3D Imaging: Volume and area of post-injection swelling were measured using both caliper measurement and 3D camera image analysis. For caliper measurements a digital caliper was utilized to measure length, width and height of the bleb that formed post-injection. The length and width are defined as the edge to edge measurements of the bleb (i.e., diameter) along their longest axes. These values were manually recorded, and the volume determined using the formula for half of an ellipsoid Vol =(2 / 3)*7r*A*B*C where A = Length / 2, B = Width / 2 and C = Height. 3D imaging was applied as a longitudinal methodology to measure post-injection swelling.RESULTS AND DISCUSSION

[0223] Pre and Post-Injection Quantitative Measurements. Quantitative measurements included applied force, back-leakage, bleb size (length, width & height) and pre and post-injection temperatures (as described in Section 7.2.

[0224] Assessment of Applied Force During Injection. The applied force was measured during the SC injection by attaching a subminiature load cell to the end of the 20-cc syringe barrel. Because the applied force for AID #1186R had a peak applied force reading of 238N, this was assessed as an outlier in the data set (Grubb’s test). In addition, examination of the injection site indicated that part of the test solution was delivered intradermally. Consequently, the data from this animal was excluded from all analyses. Applied forces for each test solution and flow rate are summarized in Table 21 and Fig. 37. Applied force during injection for individual animals at each flow rate is shown in Fig. 38. Table 21 : Summary of Applied Forces During Injection

[0225] Assessment of Post-Injection Back-Leakage. The amount of back-leakage for each injection was measured by collecting post-injection fluid at the site using a surgical eye spear. Back- leakage for Ig-120 alone and Ig-120 + rHuPH20 are shown in Table 22 and individual animal data with Mean ± SEM is shown in Fig. 39.Table 22: Mean Weight of Back-leakage (mg ± SEM)

[0226] Assessment of Post-Injection Bleb Volume, Area, and Height (Caliper Measurements) . The local injection site swelling was marked and measured using a digital caliper. Bleb volume, dispersion area and swelling height of each bleb was determined as described in Section 7.2 and are summarized in Table 23 for Ig-120 and Ig-120 + rHuPH20. Individual post-injection bleb volume, area and height values are shown in Figs. 40-42.Table 23: Bleb Volume, Area and Height After Injection of Ig-120 + rHuPH20 Using Caliper Measurement (Mean ± SEM)

[0227] Assessment of Post-Injection Bleb Shape, Volume, Area, and Height (3D Imaging). Pre- and post-injection photographs were taken using a 3D imaging system (Canfield Scientific). Colorimetric surface contour maps of each post-injection bleb for Ig-120 and Ig-120 + rHuPH20 in Fig. 43.

[0228] Post-injection bleb volume, area and height for Ig-120 and Ig-120 + rHuPH20 calculated from the 3D images are summarized in Table 24. Individual post-injection bleb volume, area and height are shown graphically in Figs. 44-46.Table 24: Bleb Volume, Area and Height After Injection of Ig-120 + rHuPH20 Assessed Using 3D Imaging (Mean ± SEM)

[0229] Assessment of Post-Injection Temperature Changes. The temperature of the injection site was measured immediately prior to needle insertion using an infrared thermometer. It was then remeasured at the end of the injection to determine if any significant changes in temperature may occur as a result of flow rate. Temperature changes are summarized in Fig. 47. The overall temperature decrease was greater for injections of Ig-120 + rHuPH20 but was not found to be statistically significantly different than that for Ig-120 alone.Qualitative Assessment of Local Injection Sites

[0230] Following the completion of the 10 m injections the qualitative assessments for erythema, swelling size and firmness by the three different scorers was performed as described in Section 6.

[0231] Qualitative Assessment of Post-Injection Erythema. Erythema was minor for both test solutions. It was observed most frequently at the post-injection T15 timepoint but rapidly resolved in all cases with a substantial reduction by the T30 timepoint and near complete resolution by the T2h timepoint. The scoring by the three evaluators for erythema (Mean ± SEM) for each test solution isshown in Fig. 48 and summarized in Table 25. Erythema was extremely mild and transient for injections of Ig-120 and Ig-120 + rHuPH20 and rapidly resolved by 30 minutes post-injection for both groups.Table 25: Erythema Scores Post-Injection for Ig-120 and Ig-120 + rHuPH20 (Mean ± SEM)

[0232] Qualitative Assessment of Post-Injection Swelling Size. Scoring by the three evaluators for swelling size (Mean ± SEM) for each test solution over time is shown in Fig. 49 and summarized in Table 26. While initial swelling scores were similar for both Ig-120 and Ig-120 + rHuPH20, the post-injection blebs resolved more rapidly for the injections of Ig-120 + rHuPH20 and were considered resolved by T2h post-injection. In contrast the post-injection swelling was still evident for the injections of Ig-120 alone at T2h. All swelling appeared to be resolved for both groups by T24h post-injection.Table 26: Swelling Scores Post-Injection for Igl20 + rHuPH20 (Mean ± SEM)

[0233] Qualitative Assessment of Post-Injection Firmness (Induration). The hardness (induration) of the post-injection blebs were also evaluated by the independent scorers. The scoring for induration (Mean ± SEM) for each test solution over time is shown in Fig. 50 and summarized in Table 27. Induration of post-injection blebs was notably reduced for injections of Ig-120 + rHuPH20 compared to lg-120 alone. The induration was almost completely resolved by T2h for Ig-120 + rHuPH20 injections while noticeable induration was still detected for injections of Ig-120 alone at T2h. All induration appeared to be resolved for both groups by T24h post-injection.Table 27: Induration Scores Post-Injection for Igl20 + rHuPH20 (Mean ± SEM)PHOTOGRAPHIC ASSESSMENT OF INJECTION SITES POST DOSING

[0234] The injection sites were photographed before and after the 10 mL injection procedure. Photographic images are shown in Figs. 51-62. It should be noted at the 2 hour timepoint (T2h), the photos of the animal were not taken while it was anesthetized, but rather manually held by an animal technician. Because of the increased stress to the animal, this resulted in some flushing of the skin for some animals. In addition, the injection site may have had some increased tension (skin stretching) when photographed. Therefore, the qualitative scoring is considered the more accurate assessment of the injection site at the 2h timepoint.SUMMARY AND CONCLUSIONS

[0235] Applied force for injections of Ig-120 + rHuPH20 were reduced by ~8% compared to Ig- 120 alone with less variability compared to Ig-120 alone.

[0236] Back-leakage was reduced -60% for injections of Ig-120 + rHuPH20 compared to Ig-120 alone.

[0237] Post-injection swelling volume, area and bleb height were reduced for injections of Ig-120 + rHuPH20 compared to Ig-120 alone.

[0238] Qualitative assessments of blebs over time demonstrated that the swelling size and induration was reduced for all angled injections compared to previous testing using vertical needle insertion (study 22098).

[0239] The swelling and induration of blebs that formed from Ig-120 + rHuPH20 injections resolved faster than those for Ig-120 alone. Swelling size was mostly resolved (< 1) by T2h while induration was mostly resolved by thirty minutes post-injection.Example 3: Assessment of a Ten mL HVAI Injection of Ig-120 With and Without rHuPH20 Using a Low Angled (20°) Platform and 25G NeedleEXPERIMENTAL DESIGN AND METHODSTest and Control Articles

[0240] Ig-120 Test Solution (Human Gamma Globulin (HGG)Description: Clear liquid reconstituted from lyophilized powderConcentration: 120 mg / mLFormulation: 20 mM Histidine, 130 mM sodium chloride, pH 6.5Lot number: NB 1090-49 (HGG-003)Storage Conditions: <-65 °CHandling Conditions: Standard laboratory precautionsSupplier: Biomed SupplyFormulated by: Halozyme Product Development

[0241] Recombinant Human Hyaluronidase PH20 (rHuPH20)Description: Clear colorless liquidLot number: 1 -FIN-3928Concentration: 9.3 mg / mLEnzyme activity: 1,039,763 U / mLFormulation: 10 mM Histidine, 130 mM sodium chloride, pH 6.5Storage Conditions: < 70°CHandling Conditions: Standard laboratory precautionsSupplier: Halozyme Therapeutics, Inc.

[0242] Ig Dilution BufferDescription: Clear colorless liquidFormulation: 20 mM histidine, 130 mM sodium chloride, pH 6.5Batch / Lot: NB 1078- 18Storage Conditions: 2-8°CHandling Conditions: Standard laboratory precautionsSupplier: Halozyme Therapeutics, IncFormulation

[0243] Preparation of Test Solutions. Two test solutions were utilized for this study. The first test solution was Ig-120 alone. The second test solution was Ig-120 + rHuPH20. To ensure sufficient test solution for syringe filling, 90 mL of each test solution was prepared.

[0244] To prepare the co-mix of Ig-120 + rHuPH20, 0.173 mL of rHuPH20 was added to the 90 mL of Ig-120 to yield a test solution of Ig-120 + rHuPH20 with a final concentration of -2000 U / mL. After preparation of the co-mix, both test solutions were used for syringe filling.

[0245] Device Syringe Filling. Filling of all device syringes occurred in a sterile fill and finish hood under aseptic conditions. Cyclic olefin copolymer (COC) syringes were filled with 10.95 mL of either Ig-120 alone or the co-mix of Ig-120 + rHuPH20. To fill the syringes a sterile rubber plunger was initially inserted into the sterile syringe to a loading position. A 20-cc syringe was then filled withthe test solution and a syringe pump was then used to fill the COC syringe with 10.95 mL of the test solution. After adding the test solution to the syringe, the rubber plunger was brought to the final fill position to remove residual air in the syringe. The syringe was then capped and stored at 2-8°C until it was the device was assembled.

[0246] Device Assembly. All HVAIs were assembled on the day prior to use. All HVAIs were Genl devices using a 25 Ibf spring. The capped COC syringe was assembled into the HVAI using a custom jig. Immediately prior to use, the syringe was uncapped and a 25G x l-’A” needle was attached to the Luer fitting of the HVAI. The HVAI was then inserted into the 20° platform and the final needle depth measured and recorded. All HVAIs were positioned in the platform so that the needle bevel faced upwards from the plane of the animal’s skin.Animal Description

[0247] A summary of the weight of each animal on the day of treatment is provided in Table 28.

[0248] Species: Pig (Sus scrofa domestica)Strain: Yucatan MiniatureSex: FemaleAge: ~ 3 monthsQuantity: 6 animalsSource: Premier BioSource (Ramona, CA)Table 28: Summary of Animal Weights (Day of Treatment)

[0249] Husbandry. Animals were housed in steel pens with automatic water provided ad libitum. Animals were fed twice daily (AM and PM) but were kept NPO after midnight on the day of the study. The room environment was set to maintain a temperature of - 19-23°C and a relative humidity of 40-70%, with a 12 hour light / 12 hour dark time cycle. Animals were acclimated to the vivarium for a minimum of 3 days prior to study start.Test Materials

[0250] The test materials used in the study are listed in Table 29.Table 29: Summary of Test MaterialsEXPERIMENTAL DESIGN

[0251] In this study, two 10 mL injections were administered to the abdomen of a Yucatan miniature pig. On one side of the abdomen, a test solution of Ig-120 alone was administered and a second injection of Ig-120 + rHuPH20 on the contralateral side of the animal. All test solutions containing rHuPH20 were formulated at 2000 U / mL. An HVAI was used as an off-body device to deliver the test solution. The test solutions for each animal are summarized in Table 30.Table 30: Description of Treatments

[0252] The length of each injection was measured and recorded using a stopwatch. Immediately post-injection, the needle was removed and back-leakage collected for 30 seconds using a pre- weighed eye spear. Swelling size was then measured using a digital caliper and by obtaining a 3D image capture immediately post-injection (TO). Caliper measurement of bleb size was also taken and at 15 minutes (T15) and 30 minutes (T30) post-injection. Standard photographs were obtained at TO, T15, T30, T2h (2h post-injection) and T24h (24h post-injection).

[0253] Additional post-injection qualitative injection site evaluations for erythema, swelling and induration were performed at TO, T15, T30 and T2h with a final evaluation at approximately T24h post-injection. In summary, the endpoints for the study were: measurement of HVAI injection time (digital stopwatch); collection and measurement of back-leakage weight using an eye spear; caliper measurement of bleb size post-injection using digital caliper at TO, T15 and T30; measurement of bleb volume, area and height at TO using 3D imaging; assessment of blebs for erythema, swelling size and induration at times TO, T15, T30, T2h and T24h; and photographic documentation of all injection sites: pre-inj ection, TO, T15, T30, T2h and T24h.PROCEDURES

[0254] On the day prior to the study, HVAI devices (one Ig-120; one Ig-120 + rHuPH20) were withdrawn from refrigerated storage and brought to room temperature for at least 30 minutes prior to use. The first injection for each animal was always Ig-120 followed by a second injection on the contralateral side of the animal with Ig-120 + rHuPH20. Injection sites were located on the left and right abdominal regions, ~5 cm cranially from the inguinal fold towards the midline and ~3 cm towards the midline of the animal.

[0255] Prior to the start of study, animals were assessed for general health, and body weights were collected. Animals were anesthetized with isoflurane gas and placed in dorsal recumbence on a heated surgical table and were maintained under isoflurane gas for the entire duration of the procedure. The abdominal region was cleaned with Nolvasan and the injection site wiped with gauze containing 70% isopropanol and wiped dry with sterile gauze. Each of the injection sites were marked with a permanent marker and then photographed with the standard and 3D cameras prior to HVAI needle insertion. The HVAI was mounted into the platform and the needle bevel confirmed to face upward. The HVAI was unlocked and the needle cap was removed and discarded prior to needle insertion.

[0256] The skin was pinched at the injection site and the needle was inserted into the pinched skin and allowed to freely float in the SC space. After needle insertion, the injection started by pushing the activation button. The injection was timed using a digital stopwatch. Upon completion of injection, the injection time was recorded and the needle was withdrawn from the skin. Test solution back-leakage at the injection site was absorbed to a pre-weighed eye spear for 30 seconds and the weight of the eye spear was measured. The margins of the injection site bleb were then marked with a permanent marker and measured for length, width, and height using a digital caliper, then photographed with the standard and 3D cameras immediately post-injection (TO). Caliper measurements were also taken at T15 and T30 post-injection. After the second injection, two additional syringes (one with Ig-120 and one with Ig-120 + rHuPH20) were removed from refrigerated storage and brought to room temperature for use with the following animal. Local injection sites were also qualitatively assessed and graded for erythema severity, swelling size appearance, and firmness immediately after injection, using a scoring system described in Table 31, Table 32, and Table 33, respectively. Any additional observations were noted at this time. The injection site was then qualitatively assessed at TO, T15 and T30 and photographed using a standard camera while the animal was still under anesthesia. The injection sites were again qualitatively assessed and photographed at T2h but the animal was conscious and hand-held by an animal technician.

[0257] After the first injection and while the animal was still sedated, the identical procedure was repeated on the contralateral side of the animal with the second test solution (Ig-120 + rHuPH20). Thirty minutes after completion of the second injection and after the final assessments at T30 minutes, the animal was removed from anesthesia and returned to its home cage for recovery. On the following day at approximately 24 hours, the animal was re-anesthetized, and both injection sites assessed for erythema, induration and swelling, and photographed. Following the T24h assessment, the animal was humanely euthanized using an injectable euthanasia drug.Table 31 : Grading Scale for Erythema FormationTable 32: Grading Scale for Swelling Size FormationTable 33: Grading Scale for Swelling Firmness (Induration)CALCULATIONS AND STATISTICAL METHODS

[0258] Statistical Methods. Statistical comparisons for Ig-120 and Ig-120 + rHuPH20 were performed using an unpaired parametric t-test (Prism vl0.3.1, GraphPad Software, Inc., San Diego, California). All hypotheses’ tests were performed at a 5% significance level, thus considered significant if p<0.05.

[0259] Assessment of Injection Time. The time required for each inj ection was measured using a digital stopwatch and recorded after each injection.

[0260] Assessment of Swelling Volume, Area and Height Using Caliper Measurement and 3D Imaging. Volume and area of post-injection swelling were measured using both caliper measurement and 3D camera image analysis. For caliper measurements, a digital caliper was utilized to measure length, width and height of the bleb that formed post-injection. The length and width are defined as the edge to edge measurements of the bleb (i.e., diameter) along their longest axes. These values were manually recorded, and the volume determined using the formula for half of an ellipsoid Vol = (2 / 3)*TT*A*B*C where A = Length / 2, B = Width / 2 and C = Height.

[0261] 3D imaging was applied as a longitudinal methodology to measure post-injection swelling. By obtaining high definition pre- and post-injection 3D images the distances between two registered surfaces can be determined. The camera captures images using a factory calibrated bifocal imaging system to measure distance between surfaces. Surface registration was performed using a multipoint method that utilized common landmarks between the pre-inj ection image and the post-injection image. Using the proprietary software, the volume, area and height of the post-injection swelling was calculated for each injection.RESULTS AND DISCUSSIONPre-study Enzymatic Activity Testing

[0262] The co-mix of Ig-120 + rHuPH20 was prepared on the day prior to study start and the enzyme activity of the test solution was measured using an in vitro activity assay (VV-QUAL- 00675; formerly TM010). The results of this activity testing are shown in Table 34.Table 34: Summary of Pre-Study Activity TestingPost-study Enzymatic Activity Testing

[0263] Sufficient co-mix of Ig-120 + rHuPH20 was prepared so that an extra HVAI could be assembled. The test solution from this spare device was collected at the end of the study and tested for hyaluronidase activity. The results are shown in Table 35.Table 35: Summary of Post-Study Activity TestingPre- and Post-injection Quantitative Measurements

[0264] Measurement of HVAI Injection Times. The time for each injection was measured using a stopwatch and is shown in Table 36 and Fig. 63. There was approximately a 4.3% decrease in delivery time when the Ig-120 was delivered co-mixed with rHuPH20 but this was not considered significant.Table 36: Measurement of HVAI Injection Times

[0265] Measurement of Post-injection Back-leakage. The back-leakage was collected using a pre-weighed eye spear for 30 seconds post-injection, but no measurable back-leakage occurred after any injection for either cohort.

[0266] Assessment of Post-Injection Bleb Volume, Area and Height (Caliper Measurements). The local injection site swelling (bleb) was marked and measured using a digital caliper. Bleb volume dispersion area and swelling height of each bleb was determined and this data is summarized at the TO, T15 and T30 timepoints in Table 37, Table 38, and Table 39 and individual values shown in Fig. 64, Fig. 65, and Fig. 66. Both the volume and height of blebs formed after injection of Ig-120 + rHuPH20 were found to be statistically less than after injections of Ig-120 alone.Table 37: Bleb Volume After Injection of Ig-120 and Ig-120 + rHuPH20 Over Time Using Caliper Measurement (Mean ± SEM)Table 38: Bleb Area After injection of Ig-120 and Ig-120 + rHuPH20 Over Time Using Caliper Measurement (Mean ± SEM)Table 39: Bleb Height After Injection of Ig-120 and Ig-120 + rHuPH20 Over Time Using CaliperMeasurement (Mean ± SEM)

[0267] The post-injection swelling, area and height over time were also determined at T15 and T30 and the scores are shown in Fig. 67, Fig. 68, and Fig. 69, respectively.

[0268] Assessment of Post-injection Bleb Shape, Volume, Area and Height (3D Imaging). Pre- and post-injection photographs were taken using a 3D imaging system (Canfield Scientific). This technology permits point-to-point alignment of these two images through multipoint surface registration. The distance between any two points is then represented using a colorimetric surface contour map.

[0269] Each animal had a pre-inj ection 3D image taken of the injection site followed by a second image taken immediately post-injection and these images were mapped to each other using multipoint registration. These registered pre- / post-inj ection images were then used to calculate the bleb volume, height, circumference, length, and width for each bleb using proprietary software (Vectra Hl software; Canfield Sciences). Colorimetric surface contour maps of each post-injection bleb for Ig-120 and Ig-120 + rHuPH20 are shown in Fig. 70. Differences between Ig-120 and Ig-120 + rHuPH20 were not found to be statistically significantly different for the volume, area or height as reported in Table 40 and as shown in Fig. 71, Fig. 72, and Fig. 73.Table 40: Bleb Volume, Area and Height After Injection of Ig-120 and Ig-120 + rHuPH20 Assessed Using 3D Imaging (Mean ± SEM)Qualitative Assessment of Local Injection Sites

[0270] Qualitative assessments for erythema, swelling size and induration were taken immediately post-injection (TO), 15 minutes post-injection (T15), 30 minutes post-injection (T30), 2 hours post-injection (T2h) and at approximately 24 hours post-injection (T24).

[0271] Post-injection Erythema. Erythema was only observed for one animal at TO (Ig-120 + rHuPH20). The erythema was both moderate and transient and was completely resolved by 15 minutes post-injection. No other instances of erythema were observed. The post-injection erythema scores are listed in Table 41 the post-injection swelling scores at TO and through the first two hours (T0-T15-T30-T2h) are shown in Fig. 74 and Fig. 75, respectively.Table 41 : Qualitative Assessment of Post-Injection Erythema (T0-T15-T30-T2h)

[0272] Post-injection Swelling Size. Post-injection swelling size was significantly greater for Ig- 120 than for Ig-120 + rHuPH20. Scoring by three evaluators for swelling size (Mean ± SEM) are summarized in Table 42. Individual values immediately post-injection (TO) are shown in Fig. 76 and values scored over the first two hours post-injection as well as at approximately 24h post-injection are shown in Fig. 77 and Fig. 78, respectively. The post-injection swelling scores were significantly lower for Ig-120 injections that were co-mixed with rHuPH20.Table 42: Qualitative Assessment of Post-Injection Swelling Size (T0-T15-T30-T2h)

[0273] Post-injection Induration. The induration (firmness) of the post-injection blebs for Ig- 120 was found to be significantly greater than the blebs that resulted from injection of Ig-120 + rHuPH20 (p<0.05). Scoring by three evaluators for swelling size (Mean ± SEM) are summarized in Table 43. Individual values immediately post-injection (TO) are shown in Fig. 79 and values scored over time (Mean ± SEM) are shown in Fig. 80. Change in induration score over time is shown in Fig. 81.Table 43: Qualitative Assessment of Post-Injection Induration (T0-T15-T30-T2h)COMPARISON OF ULTRA-LOW ANGLED INJECTION (20°) WITH ANGLED (45°) AND VERTICAL (90°) INJECTIONS

[0274] The endpoints measured in this study (both quantitative and qualitative) were also measured in previous studies using two different injection angles. These previous studies measuredthese endpoints for vertical injections (90°; study 22098) as well as for low angled injections (45°; studies 22099 & 24075) using the same test materials (Ig-120 ± rHuPH20) with similar injections depths (~7.5 mm). This allows for direct comparison of these injection conditions to assess the impact of angle of needle insertion. Because two studies were conducted for angled injections at 45°, these results are shown both separately and as a mean of both studies.Pre- and Post-injection Quantitative Measurements (All studies)

[0275] Comparison of HVAI Injection Times - All angles. The injection times for each delivery method were compared and shown in Table 44 and Fig. 82. While the mean injection time was almost identical for 90° and 45° injections (43.4; 43.4 sec, respectively), the mean injection time for the lowest angle (20°) was -22% lower than that for the other injections.Table 44: Comparison of HVAI Injection Times - 90° - 45° - 20°

[0276] Comparison of HVAI Back-Leakage -All angles. The mean back-leakage after each injection was also compared between studies and the results are shown in Table 45 and Fig. 83. Table 45: Comparison of HVAI Back-leakage - 90° - 45° - 20°

[0277] C "omparison of HVAI Post-injection Swelling Volume -All angles. The mean postinjection swelling volume was measured using digital calipers. The results from all three angles of delivery are shown in Table 46 and Fig. 84. The post-injection swelling volume was measured over time and is shown in Fig. 85.Table 46: Comparison of Post-injection Swelling Volume - All angles

[0278] Comparison of HVAI Post-injection Swelling Area - All angles. The mean postinjection swelling area was measured using digital calipers. A comparison of the area measured in each study at TO is shown in Table 47 and Fig. 86. The mean bleb area over time is shown in Fig. 87.Table 47: Comparison of Post-injection Swelling Area Over Time (90°-45°-20°)

[0279] Comparison of HVAI Post-injection Swelling Height Over Time (90°-45°-20°). The mean post-injection swelling height was measured using a digital caliper. Comparison of the three injection angles at TO is shown in Table 48 and at TO in Fig. 88. The post-injection swelling height over time for all injections is shown in Fig. 89.Table 48: Comparison of Post-injection Swelling Height - All anglesPre- and Post-injection Qualitative Measurements (All studies)

[0280] Comparison of HVAI Post-injection Swelling Size. Qualitative endpoints from each study were also compared. The post-injection swelling scores from each study measured immediately post-injection (TO) are shown in Table 49.Table 49: Comparison of Post-Injection Swelling Scores - All angles

[0281] C Comparison of HV Al Post-injection Induration. The post-injection swelling scores were also evaluated over time. The post-injection induration scores from each study measured immediately post-injection (TO) are shown in Table 50. Table 50: Comparison of Post-Injection Induration Scores - All anglesPHOTOGRAPHS OF INJECTION SITES PRE- AND POST-INJECTION

[0282] The injection sites were photographed before and immediately post-injection and then at T15, T30, T2h and T24h. Photographic images are shown in Fig. 90 through Fig. 101.

Claims

CLAIMS1. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject via subcutaneous administration about 3 mL to about 50 mL of a formulation comprising a therapeutically effective amount of an active ingredient selected from a small molecule, a peptide fragment, a biologic, a nanoparticle, an antibody, an antibody fragment, and a small molecule antiviral; wherein the subcutaneous administration is facilitated by a subcutaneous administration system comprising a platform attachment comprising: a base having a generally flat distal surface configured to contact a subject’s skin, the base having an aperture extending therethrough; and a housing configured to receive and support an injection apparatus containing the formulation, the housing extending non-perpendicularly in a proximal direction from the base, the housing forming a conduit in fluid communication with the aperture of the base.

2. The method of claim 1, wherein the injection apparatus containing the formulation further comprises a needle extending into the housing and through the aperture of the base during use.

3. The method of claim 2, wherein the housing comprises indicia to indicate a preferred alignment of the injection apparatus relative to the housing.

4. The method of claim 3, wherein when the injection apparatus is coupled to the housing in the preferred alignment, an open portion of the needle is facing generally perpendicular to the subject’s skin.

5. The method of claim 2, wherein the injection apparatus containing the formulation is an auto-injector.

6. The method of claim 5, wherein the auto-injector contains at least 3 mL of the formulation.

7. The method of claim 2, wherein the injection apparatus containing the formulation is an injection line.

8. The method of claim 1, wherein the distal surface of the base is a generally reuleaux triangle shape.

9. The method of claim 8, wherein the distal surface of the base comprises a lip extending around a periphery and protruding distally therefrom.

10. The method of claim 8, wherein the distal surface of the base comprises one or more ridges protruding distally therefrom.

11. The method of claim 1, wherein an internal surface of the housing comprises at least one protrusion extending radially thereof.

12. The method of claim 1, wherein the housing extends at an angle of about 5 degrees, about 10 degrees, about 15 degrees, about 20 degrees, about 25 degrees, about 30 degrees, about 35 degrees, about 40 degrees, about 45 degrees, about 50 degrees, about 55 degrees, about 60 degrees, about 65 degrees, about 70 degrees, about 75 degrees, about 80 degrees, about 85 degrees, or less than about 90 degrees relative to the distal surface of the base.

13. The method of claim 1, wherein the housing extends at an angle of 45 degrees relative to the distal surface of the base.

14. The method of claim 1, wherein the conduit of the housing includes threads configured to engage corresponding threads of the injection apparatus.

15. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject via subcutaneous administration about 3 m to about 50 mL of a formulation comprising a therapeutically effective amount of an active ingredient selected from a small molecule, a peptide fragment, a biologic, a nanoparticle, an antibody, an antibody fragment, and a small molecule antiviral; wherein the subcutaneous administration is facilitated by a subcutaneous administration system comprising a platform attachment comprising: a base having a generally flat distal surface configured to contact a subject’s skin, the base having an aperture extending therethrough; anda housing configured to receive and support an injection apparatus containing the formulation, the housing extending non-perpendicularly in a proximal direction from the base, the housing forming a conduit in fluid communication with the aperture of the base, wherein the injection apparatus containing the formulation comprises a needle extending into the housing and through the aperture of the base during use.

16. The method of claim 15, wherein the base is a generally reuleaux triangle shape.

16. A method of subcutaneously administering a formulation, the method comprising: attaching to a subject’s skin a platform attachment having a housing with an aperture extending therethrough and a generally flat base angled relative to the housing; and attaching to the housing an injection apparatus containing a formulation such that a needle extends distally through the aperture and into the subject’s skin.

17. The method of claim 16, wherein attaching the injection apparatus to the housing inserts the needle subcutaneously in the subject’s skin at an angle of about 5 degrees, about 10 degrees, about 15 degrees, about 20 degrees, about 25 degrees, about 30 degrees , about 35 degrees, about 40 degrees, about 45 degrees, about 50 degrees, about 55 degrees, about 60 degrees, about 65 degrees, about 70 degrees, about 75 degrees, about 80 degrees, about 85 degrees, or less than about 90 degrees relative to the base.

18. The method of claim 17 further comprising: delivering about 3 m to about 50 mL of the formulation through the needle.

19. The method of claim 16, wherein attaching the injection apparatus to the housing comprises attaching an auto-injector to the housing.

20. The method of claim 16, wherein attaching the injection apparatus to the housing comprises attaching an injection line to the housing.

21. The method of any one of claim 1-20, wherein the formulation further comprises a hyaluronidase enzyme.

22. The method of any one of claims 1-21, wherein the hyaluronidase enzyme is a recombinant human hyaluronidase enzyme.

23. The method of any one of claims 1-22, wherein the hyaluronidase enzyme is recombinant human hyaluronidase PH20 enzyme.

24. The method of any one of claims 1-23, wherein the hyaluronidase enzyme has an activity of about 150 U / mL to about 150 kU / mL.

25. The method of any one of claims 1-23, wherein the hyaluronidase enzyme has an activity of about 500 U / mL to about 5,000 U / mL.

26. The method of any one of claims 1-23, wherein the hyaluronidase enzyme has an activity of about 1,500 U / mL to about 10,000 U / mL.

27. The method of any one of claims 1-26, wherein the active ingredient is a small molecule, a peptide fragment, a biologic, or a nanoparticle.

28. The method of any one of claims 1-27, wherein the active ingredient is an antibody, an antibody fragment, or a small molecule antiviral.

29. The method of any one of claims 1-28, comprising administering to the subject about 10 mL to about 20 mL of the formulation.

30. The method of any one of claims 1-29, comprising administering to the subject about 3 mb to about 25 mL of the formulation.

31. The method of any one of claims 1-30, comprising administering to the subject about 3 mL, about 3.1 mL, about 3.2 mL, about 3.4 mL, about 3.5 mL, about 3.6 mL, about 3.7 mL, about 3.8 mL, about 3.9 mL, about 4 mL, about 4.1 mL, about 4.2 mL, about 4.3 mL, about 4.4 mL, about 4.5 mL, about 5 mL, about 5.1 mL, about 5.2 mL, about 5.3 mL, about 5.4 mL, about 5.5 mL, about 5.6 mL, about 5.7 mL, about 5.8 mL, about 5.9 mL, about 6 mL, about 6.1 mL, about 6.2 mL, about 6.3 mL, about 6.4 mL, about 6.5 mL, about 6.6 mL, about 6.7 mL, about 6.8 mL, about 6.9 mL, about 7 mL, about 7.1 mL, about 7.2 mL, about 7.3 mL about 7.4 mL, about 7.5 mL, about 7.6 mL, about 7.7 mL, about 7.8 mL, about 7.9 mL, about 8 mL, about 8.1 mL, about 8.2 mL, about 8.3 mL, about8.4 mL, about 8.5 mL, about 8.6 mL, about 8.7 mL, about 8.8 mL, about 8.9 mL, about 9 mL, about9.1 mL, about 9.2 mL, about 9.3 mL, about 9.4 mL, about 9.5 mL, about 9.6 mL, about 9.7 mL, about 9.8 mL, about 9.9 mL, about 10 mL, about 10.1 mL, about 10.2 mL, about 10.3 mL, about10.4 mL, about 10.5 mL, about 10.6 mL, about 10.7 mL, about 10.8 mL, about 10.9 mL, about 11 mL, about 11.1 mL, about 11.2 mL, about 11.3 mL, about 11.4 mL, about 11.5 mL, about 11.6 mL, about 11.7 mL, about 11.8 mL, about 11.9 mL, about 12 mL, about 12.1 mL, about 12.2 mL, about 12.3 mL, about 12.4 mL, about 12.5 mL, about 12.6 mL, about 12.7 mL, about 12.8 mL, about 12.9 mL, about 13 mL, about 13.1 mL, about 13.2 mL, about 13.3 mL, about 13.4 mL, about 13.5 mL, about 13.6 mL, about 13.7 mL, about 13.8 mL, about 13.9 mL, about 14 mL, about 14.1 mL, about 14.2 mL, about 14.3 mL, about 14.4 mL, about 14.5 mL, about 14.6 mL, about 14.7 mL, about 14.8 mL, about 14.9 mL, about 15 mL, about 15.1 mL, about 15.2 mL, about 15.3 mL, about 15.4 mL, about 15.5 mL, about 15.6 mL, about 15.7 mL, about 15.8 mL, about 15.9 mL, about 16 mL, about16.1 mL, about 16.2 mL, about 16.3 mL, about 16.4 mL, about 16.5 mL, about 16.6 mL, about 16.7 mL, about 16.8 mL, about 16.9 mL about 17 mL, about 17.1 mL, about 17.2 mL, about 17.3 mL, about 17.4 mL, about 17.5 mL, about 17.6 mL, about 17.7 mL, about 17.8 mL, about 17.9 mL, about 18 mL, about 18.1 mL, about 18.2 mL, about 18.3 mL, about 18.4 mL, about 18.5 mL, about 18.6 mL, about 18.7 mL, about 18.8 mL, about 18.9 mL, about 19 mL, about 19.1 mL, about 19.2 mL, about 19.3 mL, about 19.4 mL, about 19.5 mL, about 19.6 mL, about 19.7 mL, about 19.8 mL, about 19.9 mL, about 20 mL, about 20.1 mL, about 20.2 mL, about 20.3 mL, about 20.4 mL, about20.5 mL, about 20.6 mL, about 20.7 mL, about 20.8 mL, about 20.9 mL, about 21 mL, about 21.1 mL, about 21.2 mL, about 21.3 mL, about 21.4 mL, about 21.5 mL, about 21.6 mL, about 21.7 mL,about 21.8 mL, about 21.9 mL, about 22 m , about 22.1 mb, about 22.2 mb, about 22.3 mb, about 22.4 mL, about 22.5 mL, about 22.6 mL, about 22.7 mL, about 22.8 mL, about 22.9 mL, about 23 mL, about 23.1 mL, about 23.2 mL, about 23.3 mL, about 23.4 mL, about 23.5 mL, about 23.6 mL, about 23.7 mL, about 23.8 mL, about 23.9 mL, about 24 mL, about 24.1 mL, about 24.2 mL, about 24.3 mL, about 24.4 mL, about 24.5 mL, about 24.6 mL, about 24.7 mL, about 24.8 mL, about 24.9 mL, or about 25 mL.

32. The method of any one of claims 1-31, comprising administering the formulation using a high volume autoinjector.

33. The method of any one of claims 1-32, comprising administering the formulation with a starting delivery force of about 5 Ibf to about 50 Ibf using a high volume autoinjector.

34. The method of any one of claims 1-33, comprising administering the formulation with an ending delivery force of about 3 Ibf to about 20 Ibf using a high volume autoinjector.

35. The method of any one of claims 1-34, comprising administering the formulation with a starting pressure of about 50 psi to about 200 psi using a high volume autoinjector.

36. The method of any one of claims 1-34, comprising administering the formulation with an ending pressure of about 20 psi to about 75 psi using a high volume autoinjector.37 The method of any one of claims 1-36, wherein the formulation is in a prefilled syringe.

38. The method of claim 37, wherein the prefilled syringe contains about 3 mL, about 3.1 mL, about 3.2 mL, about 3.4 mL, about 3.5 mL, about 3.6 mL, about 3.7 mL, about 3.8 mL, about 3.9 mL, about 4 mL, about 4.1 mL, about 4.2 mL, about 4.3 mL, about 4.4 mL, about 4.5 mL, about 5 mL, about 5.1 mL, about 5.2 mL, about 5.3 mL, about 5.4 mL, about 5.5 mL, about 5.6 mL, about 5.7 mL, about 5.8 mL, about 5.9 mL, about 6 mL, about 6.1 mL, about 6.2 mL, about 6.3 mL, about6.4 mL, about 6.5 mL, about 6.6 mb, about 6.7 mL, about 6.8 mb, about 6.9 mb, about 7 mL, about7.1 mL, about 7.2 mL, about 7.3 mL about 7.4 mL, about 7.5 mL, about 7.6 mL, about 7.7 mL, about 7.8 mL, about 7.9 mL, about 8 mL, about 8.1 mL, about 8.2 mL, about 8.3 mL, about 8.4 mL, about 8.5 mL, about 8.6 mL, about 8.7 mL, about 8.8 mL, about 8.9 mL, about 9 mL, about 9.1 mL, about 9.2 mL, about 9.3 mL, about 9.4 mL, about 9.5 mL, about 9.6 mL, about 9.7 mL, about 9.8 mL, about 9.9 mL, about 10 mL, about 10.1 mL, about 10.2 mL, about 10.3 mL, about 10.4 mL, about 10.5 mL, about 10.6 mL, about 10.7 mL, about 10.8 mL, about 10.9 mL, about 11 mL, about11.1 mL, about 11.2 mL, about 11.3 mL, about 11.4 mL, about 11.5 mL, about 11.6 mL, about 11.7 mL, about 11.8 mL, about 11.9 mL, about 12 mL, about 12.1 mL, about 12.2 mL, about 12.3 mL, about 12.4 mL, about 12.5 mL, about 12.6 mL, about 12.7 mL, about 12.8 mL, about 12.9 mL, about 13 mL, about 13.1 mL, about 13.2 mL, about 13.3 mL, about 13.4 mL, about 13.5 mL, about 13.6 mL, about 13.7 mL, about 13.8 mL, about 13.9 mL, about 14 mL, about 14.1 mL, about 14.2 mL, about 14.3 mL, about 14.4 mL, about 14.5 mL, about 14.6 mL, about 14.7 mL, about 14.8 mL, about 14.9 mL, about 15 mL, about 15.1 mL, about 15.2 mL, about 15.3 mL, about 15.4 mL, about 15.5 mL, about 15.6 mL, about 15.7 mL, about 15.8 mL, about 15.9 mL, about 16 mL, about 16.1 mL, about 16.2 mL, about 16.3 mL, about 16.4 mL, about 16.5 mL, about 16.6 mL, about 16.7 mL, about 16.8 mL, about 16.9 mL about 17 mL, about 17.1 mL, about 17.2 mL, about 17.3 mL, about17.4 mL, about 17.5 mL, about 17.6 mL, about 17.7 mL, about 17.8 mL, about 17.9 mL, about 18 mL, about 18.1 mL, about 18.2 mL, about 18.3 mL, about 18.4 mL, about 18.5 mL, about 18.6 mL, about 18.7 mL, about 18.8 mL, about 18.9 mL, about 19 mL, about 19.1 mL, about 19.2 mL, about 19.3 mL, about 19.4 mL, about 19.5 mL, about 19.6 mL, about 19.7 mL, about 19.8 mL, about 19.9 mL, about 20 mL, about 20.1 mL, about 20.2 mL, about 20.3 mL, about 20.4 mL, about 20.5 mL, about 20.6 mL, about 20.7 mL, about 20.8 mL, about 20.9 mL, about 21 mL, about 21.1 mL, about21.2 mL, about 21.3 mL, about 21.4 mL, about 21.5 mL, about 21.6 mL, about 21.7 mL, about 21.8 mL, about 21.9 mL, about 22 mL, about 22.1 mL, about 22.2 mL, about 22.3 mL, about 22.4 mL, about 22.5 mL, about 22.6 mL, about 22.7 mL, about 22.8 mL, about 22.9 mL, about 23 mL, about 23.1 mL, about 23.2 mL, about 23.3 mL, about 23.4 mL, about 23.5 mL, about 23.6 mL, about 23.7 mL, about 23.8 mL, about 23.9 mL, about 24 mL, about 24.1 mL, about 24.2 mL, about 24.3 mL, about 24.4 mL, about 24.5 mL, about 24.6 mL, about 24.7 mL, about 24.8 mL, about 24.9 mL, or about 25 mL of the formulation.

39. The method of claim 37 or 38, wherein the prefilled syringe comprises a needle having a gauge of about 20 to about 33.

40. The method of any one of claims 37-39, wherein the prefilled syringe comprises a 20 gauge needle, a 21 gauge needle, a 22 gauge needle, a 23 gauge needle, a 24 gauge needle, a 25 gauge needle, a 26 gauge needle, a 27 gauge needle, a 28 gauge needle, a 29 gauge needle, a 30 gauge needle, a 31 gauge needle, a 32 gauge needle, or a 33 gauge needle.

41. The method of any one of claims 1-40, comprising administering the formulation at a rate of about 0.08 to about 1.00 mL / sec.

42. The method of any one of claims 1-41, comprising administering the formulation at a rate of at least about 0.08 to about 1.0 mL / sec.43 The method of any one of claims 1-42, comprising administering the formulation at a rate of at least or faster than about 0.08 to about 1.00 mL / sec.

44. The method of any one of claims 1-43, wherein the administration takes about 10 seconds to about 40 seconds.

45. The method of any one of claims 1-43, wherein the administration takes at least about 10 seconds to about 40 seconds.

46. The method of any one of claims 1-43, wherein the administration takes at least or less than about 10 seconds to about 40 seconds.

47. The method of any one of claims 1-43, wherein the administration takes about 15 seconds to about 30 seconds.

48. The method of any one of claims 1-43, wherein the administration takes at least about 15 seconds to about 30 seconds.

49. The method of any one of claims 1-43, wherein the administration takes at least or less than about 15 seconds to about 30 seconds.

50. The method of any one of claims 1-43, comprising administering about 5 mL of the formulation at a rate of about 0.14 mL / sec to about 0.21 mL / sec.

51. The method any one of claims 1-43, comprising administering about 10 mL of the formulation at a rate of about 0.32 mL / sec to about 0.42 mL / sec.

52. The method of any one of claims 1-51 wherein the formulation has a viscosity of about 1 cP to about 50 cP.

53. The method of any one of claims 1-53, wherein administration of the formulation requires less applied force when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

54. The method of any one of claims 1-52, comprising administering about 5 mL of the formulation at a rate of about 0.14 mL / sec to about 0.21 mL / sec with an applied force of about 10 N to about 45 N.

55. The method of claim 54, comprising administering the formulation to the subject using a prefilled syringe comprising a 25 gauge needle.

56. The method of any one of claims 1-55, comprising administering about 10 mL of the formulation to the subject at a rate of about 0.32 mL / sec to about 0.42 mL / sec with an applied force of about 25 N to about 50 N.

57. The method of claim 56, comprising administering the formulation to the subject using a prefilled syringe comprising a 25 gauge needle.

58. The method of any one of claims 1-57, wherein administration of the formulation is faster when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

59. The method of any one of claims 1-58, wherein administration of the formulation causes fewer side effects in the subject when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

60. The method of any one of claims 1-59, wherein administration of the formulation causes less pain and discomfort in the subject when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

61. The method of any one of claims 1-60, wherein administration of the formulation causes less back leakage at the injection site when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

62. The method of claim 61, wherein the back leakage at the injection site is about 85% to about 30% less when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

63. The method of any one of claims 1-62, wherein administration of the formulation causes less swelling volume and / or swelling height at the injection site when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

64. The method of claim 63, wherein the formulation causes about 35% to about 5% less swelling and / or swelling height the injection site when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

65. The method of any one of claims 1-64, wherein administration of the formulation yields a lower bleb swelling size, less bleb induration, and / or quicker bleb resolution when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

66. The method of any one of claims 1-65, wherein administration of the formulation yields more consistent delivery times when compared to a similar formulation that does not comprise a hyaluronidase enzyme.

67. The method of any one of claims 1-66, wherein the subject is human.

68. The method of any one of claims 1-67, wherein the administering comprises the subject selfadministering the formulation.

69. The method of any one of claims 1-68, wherein the administering comprises a healthcare provider or a caregiver administering the formulation to the subject.

70. The method of any one of claims 1-69, wherein the subcutaneous administration comprises a single injection.

71. The method of any one of claims 1-70, wherein the subcutaneous administration comprises two or more injections.

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