Combined administration of gamma-globulin and hyaluronidase for determining the time of injection

Abstract

The present disclosure provides a method of determining one or more parameters of a formulation injection in a human subject, the method comprising: administering about 3 mL to about 50 mL of a formulation comprising a therapeutically effective amount of a hyaluronidase from a large volume auto-injector to an animal subject via a subcutaneous injection; measuring one or more parameters of the subcutaneous injection in the animal subject; and determining the same one or more parameters of a subcutaneous injection in a human subject from the one or more measured parameters from the animal subject; wherein the one or more parameters are selected from the group consisting of injection time, injection force, and injection site reaction data.

Description

Technical Field

[0001] The invention herein relates to compositions and combination dosing regimens, including administration of gamma globulin and hyaluronidase to assess clinical injection timing.

[0002] Cross-reference to related applications

[0003] This international application claims priority to U.S. Provisional Patent Application No. 63 / 573,391, filed April 2, 2024, and U.S. Provisional Patent Application No. 63 / 691,488, filed September 6, 2024, each of which is incorporated herein by reference in its entirety.

[0004] The sequence list is incorporated by reference.

[0005] An electronic version of the sequence list was submitted along with the document, its contents incorporated in their entirety by reference. The electronic file was created on March 31, 2025, is 101 kilobytes in size, and has the title 063995-5125WO.xml. Background Technology

[0006] Patient surveys have shown that most patients prefer long-acting injectable regimens with fewer intervals to intravenous injections.

[0007] Long-acting injectable therapies are limited by patient experience and side effects. There is a desire to obtain injectable suspensions with high concentrations of biologics to allow for less frequent administration and overcome treatment non-compliance issues, while maintaining product stability and avoiding patient side effects. Summary of the Invention

[0008] In one aspect, this disclosure provides a method for determining one or more parameters of a formulation injection in a human subject, the method comprising: administering about 3 mL to about 50 mL of a formulation containing a therapeutically effective amount of hyaluronidase to an animal subject via subcutaneous injection from a large-volume autoinjector; measuring one or more parameters of the subcutaneous injection in the animal subject; and determining the same one or more parameters of the subcutaneous injection in a human subject based on one or more measured parameters from the animal subject; wherein the one or more parameters are selected from injection time, injection force, and injection site reaction data. In one embodiment, the hyaluronidase is a soluble hyaluronidase. In one embodiment, the hyaluronidase is a recombinant human hyaluronidase. In one embodiment, the hyaluronidase is a recombinant human hyaluronidase PH20 enzyme. In one embodiment, the recombinant human hyaluronidase PH20 enzyme comprises a sequence having at least 95% amino acid sequence identity with the amino acid sequence of SEQ ID NO: 1. In one embodiment, the formulation further comprises a therapeutically effective amount of an active ingredient selected from small molecules, proteins, peptide fragments, biological agents, nanoparticles, antibodies, antibody fragments, and small molecule antiviral agents. In one embodiment, the antibody comprises a monoclonal antibody or a polyclonal antibody. In one embodiment, the monoclonal or polyclonal antibody includes trastuzumab, rituximab, pertuzumab, atezolizumab, immunoglobulin, daratumumab, egamod α, nivolumab, or olizumab. In one embodiment, the protein includes gamma globulin. In one embodiment, the animal subject is a miniature pig. In one embodiment, the animal subject is a yucatan miniature pig. In one embodiment, the method includes administering about 10 mL to about 20 mL of the formulation to the animal subject. In one embodiment, the method includes administering about 10 mL to about 20 mL of the formulation to a human subject. In one embodiment, the method includes administering about 3 mL to about 15 mL of the formulation to the animal subject. In one embodiment, the method includes administering about 3 mL to about 15 mL of the formulation to a human subject. In one embodiment, the method includes administering to an animal subject approximately 3 mL, approximately 3.1 mL, approximately 3.2 mL, approximately 3.4 mL, approximately 3.5 mL, approximately 3.6 mL, approximately 3.7 mL, approximately 3.8 mL, approximately 3.9 mL, approximately 4 mL, approximately 4.1 mL, approximately 4.2 mL, approximately 4.3 mL, approximately 4.4 mL, approximately 4.5 mL, approximately 5 mL, approximately 5.1 mL, approximately 5.2 mL, approximately 5.3 mL, approximately 5.4 mL, approximately 5.5 mL, approximately 5.6 mL, approximately 5.7 mL, approximately 5.8 mL, approximately 5.9 mL, approximately 6 mL, approximately 6.1 mL, approximately 6.2 mL, approximately 6.3 mL, approximately 6.4 mL, approximately 6.5 mL, approximately 6.6 mL, approximately 6.7 mL, approximately 6.8 mL, approximately 6.9 mL, approximately 7 mL, approximately 7.1 mL, approximately 7.2 mL, approximately 7.3 mL, or approximately 7 mL.4mL, approximately 7.5mL, approximately 7.6mL, approximately 7.7mL, approximately 7.8mL, approximately 7.9mL, approximately 8mL, approximately 8.1mL, approximately 8.2mL, approximately 8.3mL, approximately 8.4mL, approximately 8.5mL, approximately 8.6mL, approximately 8.7mL, approximately 8.8mL, approximately 8.9mL, approximately 9mL, approximately 9.1mL, approximately 9.2mL, approximately 9.3mL, approximately 9.4mL, approximately 9.5mL, approximately 9.6mL, approximately 9.7mL, approximately 9.8mL, approximately 9.9mL, approximately 10mL, approximately 10.1mL, approximately 10.2mL, approximately 10.3mL, approximately 10.4mL, approximately 10.5mL, approximately 10.6mL, approximately 10.7mL, approximately 10.8mL, approximately 10. 9 mL, approximately 11 mL, approximately 11.1 mL, approximately 11.2 mL, approximately 11.3 mL, approximately 11.4 mL, approximately 11.5 mL, approximately 11.6 mL, approximately 11.7 mL, approximately 11.8 mL, approximately 11.9 mL, approximately 12 mL, approximately 12.1 mL, approximately 12.2 mL, approximately 12.3 mL, approximately 12.4 mL, approximately 12.5 mL, approximately 12.6 mL, approximately 12.7 mL, approximately 12.8 mL, approximately 12.9 mL, approximately 13 mL, approximately 13.1 mL, approximately 13.2 mL, approximately 13.3 mL, approximately 13.4 mL, approximately 13.5 mL, approximately 13.6 mL, approximately 13.7 mL, approximately 13.8 mL, approximately 13.9 mL, approximately 14 mL, approximately 14.1 mL mL, approximately 14.2 mL, approximately 14.3 mL, approximately 14.4 mL, approximately 14.5 mL, approximately 14.6 mL, approximately 14.7 mL, approximately 14.8 mL, approximately 14.9 mL, approximately 15 mL, approximately 15.1 mL, approximately 15.2 mL, approximately 15.3 mL, approximately 15.4 mL, approximately 15.5 mL, approximately 15.6 mL, approximately 15.7 mL, approximately 15.8 mL, approximately 15.9 mL, approximately 16 mL, approximately 16.1 mL, approximately 16.2 mL, approximately 16.3 mL, approximately 16.4 mL, approximately 16.5 mL, approximately 16.6 mL, approximately 16.7 mL, approximately 16.8 mL, approximately 16.9 mL, approximately 17 mL, approximately 17.1 mL, approximately 17.2 mL, approximately 17. 3mL, approximately 17.4mL, approximately 17.5mL, approximately 17.6mL, approximately 17.7mL, approximately 17.8mL, approximately 17.9mL, approximately 18mL, approximately 18.1mL, approximately 18.2mL, approximately 18.3mL, approximately 18.4mL, approximately 18.5mL, approximately 18.6mL, approximately 18.7mL, approximately 18.8mL, approximately 18.9mL, approximately 19mL, approximately 19.1mL, approximately 19.2mL, approximately 19.3mL, approximately 19.4mL, approximately 19.5mL, approximately 19.6mL, approximately 19.7mL, approximately 19.8mL, approximately 19.9mL, approximately 20mL, approximately 20.1mL, approximately 20.2mL, approximately 20.3mL, approximately 20.4mL, approximately 20.5mL, approximately 20.6mL, approximately 20.7mL, approximately 20.8mL, approximately 20.9mL, approximately 21mL, approximately 21.1mL, approximately 21.2mL, approximately 21.3mL, approximately 21.4mL, approximately 21.5mL, approximately 21.6mL, approximately 21.7mL, approximately 21.8mL, approximately 21.9mL, approximately 22mL, approximately 22.1mL, approximately 22.2mL, approximately 22.3mL, approximately 22.4mL, approximately 22.5mL, approximately 22.6mL, approximately 22.7mL, approximately 22.8 mL, approximately 22.9 mL, approximately 23 mL, approximately 23.1 mL, approximately 23.2 mL, approximately 23.3 mL, approximately 23.4 mL, approximately 23.5 mL, approximately 23.6 mL, approximately 23.7 mL, approximately 23.8 mL, approximately 23.9 mL, approximately 24 mL, approximately 24.1 mL, approximately 24.2 mL, approximately 24.3 mL, approximately 24.4 mL, approximately 24.5 mL, approximately 24.6 mL, approximately 24.7 mL, approximately 24.8 mL, approximately 24.9 mL, or approximately 25 mL. In one embodiment, the method includes administering to a human subject approximately 3 mL, approximately 3.1 mL, approximately 3.2 mL, approximately 3.4 mL, approximately 3.5 mL, approximately 3.6 mL, approximately 3.7 mL, approximately 3.8 mL, approximately 3.9 mL, approximately 4 mL, approximately 4.1 mL, approximately 4.2 mL, approximately 4.3 mL, approximately 4.4 mL, approximately 4.5 mL, approximately 5 mL, approximately 5.1 mL, approximately 5.2 mL, approximately 5.3 mL, approximately 5.4 mL, or approximately 5.5 mL. L, approximately 5.6 mL, approximately 5.7 mL, approximately 5.8 mL, approximately 5.9 mL, approximately 6 mL, approximately 6.1 mL, approximately 6.2 mL, approximately 6.3 mL, approximately 6.4 mL, approximately 6.5 mL, approximately 6.6 mL, approximately 6.7 mL, approximately 6.8 mL, approximately 6.9 mL, approximately 7 mL, approximately 7.1 mL, approximately 7.2 mL, approximately 7.3 mL, approximately 7.4 mL, approximately 7.5 mL, approximately 7.6 mL, approximately 7.7 mL, approximately 7.8 mL, approximately 7 0.9mL, approximately 8mL, approximately 8.1mL, approximately 8.2mL, approximately 8.3mL, approximately 8.4mL, approximately 8.5mL, approximately 8.6mL, approximately 8.7mL, approximately 8.8mL, approximately 8.9mL, approximately 9mL, approximately 9.1mL, approximately 9.2mL, approximately 9.3mL, approximately 9.4mL, approximately 9.5mL, approximately 9.6mL, approximately 9.7mL, approximately 9.8mL, approximately 9.9mL, approximately 10mL, approximately 10.1mL, approximately 10.2mL L, approximately 10.3 mL, approximately 10.4 mL, approximately 10.5 mL, approximately 10.6 mL, approximately 10.7 mL, approximately 10.8 mL, approximately 10.9 mL, approximately 11 mL, approximately 11.1 mL, approximately 11.2 mL, approximately 11.3 mL, approximately 11.4 mL, approximately 11.5 mL, approximately 11.6 mL, approximately 11.7 mL, approximately 11.8 mL, approximately 11.9 mL, approximately 12 mL, approximately 12.1 mL, approximately 12.2 mL, approximately 12.3mL, approximately 12.4mL, approximately 12.5mL, approximately 12.6mL, approximately 12.7mL, approximately 12.8mL, approximately 12.9mL, approximately 13mL, approximately 13.1mL, approximately 13.2mL, approximately 13.3mL, approximately 13.4mL, approximately 13.5mL, approximately 13.6mL, approximately 13.7mL, approximately 13.8mL, approximately 13.9mL, approximately 14mL, approximately 14.1mL, approximately 14.2mL, approximately 14.3mL, approximately 14.4mL, approximately 14.5mL, approximately 14.6mL, approximately 14.7mL, approximately 14.8mL, approximately 14.9mL, approximately 15mL, approximately 15.1mL, approximately 15.2mL, approximately 15.3mL, approximately 15.4mL Approximately 15.5 mL, approximately 15.6 mL, approximately 15.7 mL, approximately 15.8 mL, approximately 15.9 mL, approximately 16 mL, approximately 16.1 mL, approximately 16.2 mL, approximately 16.3 mL, approximately 16.4 mL, approximately 16.5 mL, approximately 16.6 mL, approximately 16.7 mL, approximately 16.8 mL, approximately 16.9 mL, approximately 17 mL, approximately 17.1 mL, approximately 17.2 mL, approximately 17.3 mL, approximately 17.4 mL, approximately 17.5 mL, approximately 17.6 mL, approximately 17.7 mL, approximately 17.8 mL, approximately 17.9 mL, approximately 18 mL, approximately 18.1 mL, approximately 18.2 mL, approximately 18.3 mL, approximately 18.4 mL, approximately 18.5 mL, approximately 18. 6mL, approximately 18.7mL, approximately 18.8mL, approximately 18.9mL, approximately 19mL, approximately 19.1mL, approximately 19.2mL, approximately 19.3mL, approximately 19.4mL, approximately 19.5mL, approximately 19.6mL, approximately 19.7mL, approximately 19.8mL, approximately 19.9mL, approximately 20mL, approximately 20.1mL, approximately 20.2mL, approximately 20.3mL, approximately 20.4mL, approximately 20.5mL, approximately 20.6mL, approximately 20.7mL, approximately 20.8mL, approximately 20.9mL, approximately 21mL, approximately 21.1mL, approximately 21.2mL, approximately 21.3mL, approximately 21.4mL, approximately 21.5mL, approximately 21.6mL, approximately 21.7mL Approximately 21.8 mL, approximately 21.9 mL, approximately 22 mL, approximately 22.1 mL, approximately 22.2 mL, approximately 22.3 mL, approximately 22.4 mL, approximately 22.5 mL, approximately 22.6 mL, approximately 22.7 mL, approximately 22.8 mL, approximately 22.9 mL, approximately 23 mL, approximately 23.1 mL, approximately 23.2 mL, approximately 23.3 mL, approximately 23.4 mL, approximately 23.5 mL, approximately 23.6 mL, approximately 23.7 mL, approximately 23.8 mL, approximately 23.9 mL, approximately 24 mL, approximately 24.1 mL, approximately 24.2 mL, approximately 24.3 mL, approximately 24.4 mL, approximately 24.5 mL, approximately 24.6 mL, approximately 24.7 mL, approximately 24.8 mL, approximately 24.9 mL, or about 25 mL. In one embodiment, the formulation is in a pre-filled syringe. In one embodiment, the pre-filled 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, about 6.4 mL, about 6.5 mL, about 6.6 mL, about 6.7 mL, or about 25 mL. 6.8mL, approximately 6.9mL, approximately 7mL, approximately 7.1mL, approximately 7.2mL, approximately 7.3mL, approximately 7.4mL, approximately 7.5mL, approximately 7.6mL, approximately 7.7mL, approximately 7.8mL, approximately 7.9mL, approximately 8mL, approximately 8.1mL, approximately 8.2mL, approximately 8.3mL, approximately 8.4mL, approximately 8.5mL, approximately 8.6mL, approximately 8.7mL, approximately 8.8mL, approximately 8.9mL, approximately 9mL, approximately 9mL, approximately 9.1mL, approximately 9.2mL, approximately 9.3mL, approximately 9.4mL, approximately 9.5mL, approximately 9.6mL, approximately 9.7mL, approximately 9.8mL, approximately 9.9mL, approximately 10mL, approximately 10.1mL, approximately 10.2mL, approximately 10 0.3mL, approximately 10.4mL, approximately 10.5mL, approximately 10.6mL, approximately 10.7mL, approximately 10.8mL, approximately 10.9mL, approximately 11mL, approximately 11.1mL, approximately 11.2mL, approximately 11.3mL, approximately 11.4mL, approximately 11.5mL, approximately 11.6mL, approximately 11.7mL, approximately 11.8mL, approximately 11.9mL, approximately 12mL, approximately 12.1mL, approximately 12.2mL, approximately 12.3mL, approximately 12.4mL, approximately 12.5mL, approximately 12.6mL, approximately 12.7mL, approximately 12.8mL, approximately 12.9mL, approximately 13mL, approximately 13.1mL, approximately 13.2mL, approximately 13.3mL, approximately 13... 0.4mL, approximately 13.5mL, approximately 13.6mL, approximately 13.7mL, approximately 13.8mL, approximately 13.9mL, approximately 14mL, approximately 14.1mL, approximately 14.2mL, approximately 14.3mL, approximately 14.4mL, approximately 14.5mL, approximately 14.6mL, approximately 14.7mL, approximately 14.8mL, approximately 14.9mL, approximately 15mL, approximately 15.1mL, approximately 15.2mL, approximately 15.3mL, approximately 15.4mL, approximately 15.5mL, approximately 15.6mL, approximately 15.7mL, approximately 15.8mL, approximately 15.9mL, approximately 16mL, approximately 16.1mL, approximately 16.2mL, approximately 16.3mL, approximately 16.4mL, approximately 16.5mL, approximately 16.6mL, approximately 16.7mL, approximately 16.8mL, approximately 16.9mL, approximately 17mL, approximately 17.1mL, approximately 17.2mL, approximately 17.3mL, approximately 17.4mL, approximately 17.5mL, approximately 17.6mL, approximately 17.7mL, approximately 17.8mL, approximately 17.9mL, approximately 18mL, approximately 18.1mL, approximately 18.2mL, approximately 18.3mL, approximately 18.4mL, approximately 18.5mL, approximately 18.6mL Approximately 18.7 mL, approximately 18.8 mL, approximately 18.9 mL, approximately 19 mL, approximately 19.1 mL, approximately 19.2 mL, approximately 19.3 mL, approximately 19.4 mL, approximately 19.5 mL, approximately 19.6 mL, approximately 19.7 mL, approximately 19.8 mL, approximately 19.9 mL, approximately 20 mL, approximately 20.1 mL, approximately 20.2 mL, approximately 20.3 mL, approximately 20.4 mL, approximately 20.5 mL, approximately 20.6 mL, approximately 20.7 mL, approximately 2 0.8 mL, approximately 20.9 mL, approximately 21 mL, approximately 21.1 mL, approximately 21.2 mL, approximately 21.3 mL, approximately 21.4 mL, approximately 21.5 mL, approximately 21.6 mL, approximately 21.7 mL, approximately 21.8 mL, approximately 21.9 mL, approximately 22 mL, approximately 22.1 mL, approximately 22.2 mL, approximately 22.3 mL, approximately 22.4 mL, approximately 22.5 mL, approximately 22.6 mL, approximately 22.7 mL, approximately 22.8 mL, approximately 22.9 mL The formulation comprises approximately 23 mL, 23.1 mL, 23.2 mL, 23.3 mL, 23.4 mL, 23.5 mL, 23.6 mL, 23.7 mL, 23.8 mL, 23.9 mL, 24 mL, 24.1 mL, 24.2 mL, 24.3 mL, 24.4 mL, 24.5 mL, 24.6 mL, 24.7 mL, 24.8 mL, 24.9 mL, or 25 mL. In one embodiment, the pre-filled syringe includes a needle with a gauge of approximately 20 to approximately 33. In one embodiment, the pre-filled syringe includes 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. In one embodiment, the method includes administering the formulation to an animal subject at a rate of about 0.08 mL / s to about 1.00 mL / s. In one embodiment, the method includes administering the formulation to a human subject at a rate of about 0.08 mL / s to about 1.00 mL / s. In one embodiment, the method includes administering the formulation to an animal subject at a rate of at least about 0.08 mL / s to about 1.0 mL / s. In one embodiment, the method includes administering the formulation at a rate of at least about 0.08 mL / s to about 1.0 mL / s.The method involves administering the formulation to a human subject at a rate of 0 mL / s. In one embodiment, the method includes administering the formulation to an animal subject at a rate of at least or faster than about 0.08 mL / s to about 1.00 mL / s. In one embodiment, the method includes administering the formulation to a human subject at a rate of at least or faster than about 0.08 mL / s to about 1.00 mL / s. In one embodiment, the administration of the formulation to the animal subject takes about 10 seconds to about 40 seconds. In one embodiment, the administration of the formulation to the human subject takes about 10 seconds to about 40 seconds. In one embodiment, the administration of the formulation to the animal subject takes at least about 10 seconds to about 40 seconds. In one embodiment, the administration of the formulation to the human subject takes at least about 10 seconds to about 40 seconds. In one embodiment, the administration of the formulation to the animal subject takes at least or less than about 10 seconds to about 40 seconds. In one embodiment, the administration of the formulation to the human subject takes at least or less than about 10 seconds to about 40 seconds. In one embodiment, the administration of the formulation to the animal subject takes about 15 seconds to about 30 seconds. In one embodiment, the administration of the formulation to the human subject takes about 15 seconds to about 30 seconds. In one embodiment, administration of the formulation to an animal subject takes at least about 15 seconds to about 30 seconds. In one embodiment, administration of the formulation to a human subject takes at least about 15 seconds to about 30 seconds. In one embodiment, administration of the formulation to an animal subject takes at least or less than about 15 seconds to about 30 seconds. In one embodiment, administration of the formulation to a human subject takes at least or less than about 15 seconds to about 30 seconds. In one embodiment, the method includes administering about 10 mL of the formulation to an animal subject at a rate of about 0.32 mL / second to about 0.42 mL / second. In one embodiment, the method includes administering about 10 mL of the formulation to a human subject at a rate of about 0.32 mL / second to about 0.42 mL / second.

[0009] In one aspect, this disclosure provides a method for determining one or more parameters of a formulation injection in a human subject, the method comprising: administering about 3 mL to about 50 mL of a formulation containing a therapeutically effective amount of hyaluronidase to an animal subject via subcutaneous injection from a large-volume autoinjector; measuring one or more parameters of the subcutaneous injection in the animal subject; and determining the same one or more parameters of the subcutaneous injection in the human subject based on one or more measured parameters from the animal subject; wherein the one or more parameters are injection force. In one embodiment, the animal subject is a miniature pig. In one embodiment, the animal subject is a Yucatan miniature pig. In one embodiment, the formulation further comprises gamma globulin. In one embodiment, subcutaneous administration to the animal subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, subcutaneous administration to the human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, the method comprises determining the injection force of the subcutaneous injection in the human subject by multiplying the measured injection force of the animal subject by a scaling factor. In one embodiment, the scaling factor is from about 0.80 to about 0.99. In one embodiment, the scaling factor is from about 0.85 to about 0.95; optionally, from about 0.88 to about 0.92. In one embodiment, the injection force is measured in Newtons (N). In one embodiment, the method includes: administering about 10 mL of the formulation to a miniature pig animal subject via subcutaneous injection at a rate of about 0.30 mL / s to about 0.42 mL / s, wherein a large-volume autoinjector dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle; measuring the injection force of the subcutaneous injection in the miniature pig from about 25 N to about 50 N; and determining the injection force of the same subcutaneous injection in a human subject by multiplying the miniature pig injection force by a scaling factor of about 0.85 to about 0.95, thereby obtaining an estimated injection force of about 21 N to about 48 N. In one embodiment, the method includes: administering about 10 mL of the formulation to a miniature pig animal subject via subcutaneous injection at a rate of about 0.30 mL / s to about 0.36 mL / s, wherein a large-volume autoinjector dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle; measuring the injection force of the subcutaneous injection in the miniature pig as about 40 N to about 46 N; and determining the injection force of the same subcutaneous injection in a human subject by multiplying the injection force in the miniature pig by a scaling factor of about 0.88 to about 0.92, thereby obtaining an estimated injection force of about 35 N to about 42 N. In one embodiment, the subcutaneous administration to the animal subject comprises a single injection. In one embodiment, the subcutaneous administration to the human subject comprises a single injection.In one embodiment, subcutaneous administration to an animal subject comprises two or more injections. In one embodiment, subcutaneous administration to a human subject comprises two or more injections.

[0010] In one aspect, this disclosure provides a method for determining one or more parameters of a formulation injection in a human subject, the method comprising: administering about 3 mL to about 50 mL of a formulation containing a therapeutically effective amount of hyaluronidase to an animal subject via subcutaneous injection from a large-volume autoinjector; measuring one or more parameters of the subcutaneous injection in the animal subject; and determining the same one or more parameters of the subcutaneous injection in the human subject based on one or more measured parameters from the animal subject; wherein one or more parameters are injection time. In one embodiment, the animal subject is a miniature pig. In one embodiment, the animal subject is a Yucatan miniature pig. In one embodiment, the formulation further comprises gamma globulin. In one embodiment, subcutaneous administration to the animal subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, subcutaneous administration to the human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, the method includes determining the injection time of the subcutaneous injection in the human subject by multiplying the measured injection time of the animal subject by a scaling factor. In one embodiment, the scaling factor is from about 0.80 to about 0.99. In one embodiment, the scaling factor is from about 0.85 to about 0.95; optionally, from about 0.88 to about 0.92. In one embodiment, the injection time is measured in seconds. In one embodiment, the method includes: administering about 10 mL of the formulation to a miniature pig animal subject via subcutaneous injection at a rate of about 0.30 mL / s to about 0.36 mL / s, wherein a large-volume autoinjector dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle; measuring the injection time of the subcutaneous injection in the miniature pig from about 15 seconds to about 35 seconds; and determining the injection time of the same subcutaneous injection in a human subject by multiplying the injection time in the miniature pig by a scaling factor of about 0.85 to about 0.95, thereby obtaining an estimated injection time of about 13 seconds to about 33 seconds. In one embodiment, the method includes: administering about 10 mL of the formulation to a miniature pig animal subject via subcutaneous injection at a rate of about 0.30 mL / s to about 0.36 mL / s, wherein a large-volume autoinjector dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle; measuring the injection time of the subcutaneous injection in the miniature pig from about 18 seconds to about 30 seconds; and determining the injection time of the same subcutaneous injection in a human subject by multiplying the injection time in the miniature pig by a scaling factor of about 0.88 to about 0.92, thereby obtaining an estimated injection time of about 16 seconds to about 28 seconds. In one embodiment, the subcutaneous administration to the animal subject comprises a single injection. In one embodiment, the subcutaneous administration to the human subject comprises a single injection.In one embodiment, subcutaneous administration to an animal subject comprises two or more injections. In one embodiment, subcutaneous administration to a human subject comprises two or more injections.

[0011] In one aspect, this disclosure provides a method for determining one or more parameters of a formulation injection in a human subject, the method comprising: administering about 3 mL to about 50 mL of a formulation containing a therapeutically effective amount of hyaluronidase to an animal subject via subcutaneous injection from a large-volume autoinjector; measuring one or more parameters of the subcutaneous injection in the animal subject; and determining the same one or more parameters of the subcutaneous injection in the human subject based on one or more measured parameters from the animal subject; wherein the one or more parameters are injection site reaction data. In one embodiment, the injection site reaction data includes one or more of backleakage at the injection site, swelling volume at the injection site, swelling height at the injection site, wheal size at the injection site, wheal induration at the injection site, and wheal regression at the injection site. In one embodiment, the animal subject is a miniature pig. In one embodiment, the animal subject is a Yucatan miniature pig. In one embodiment, the formulation further comprises gamma globulin. In one embodiment, subcutaneous administration to the animal subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, the method includes: administering approximately 10 mL of the formulation to a miniature pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.42 mL / s, wherein the large-volume autoinjector dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle; measuring the response at one or more injection sites in the miniature pig and assigning a score of 0, 1, 2, 3, or 4; and determining that the responses at one or more injection sites in the same subcutaneous injection in a human subject will have the same score. In one embodiment, the method includes: administering about 10 mL of the formulation to a miniature pig animal subject via subcutaneous injection at a rate of about 0.30 mL / s to about 0.36 mL / s, wherein a large-volume autoinjector dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle; measuring the response at one or more injection sites in the miniature pig via subcutaneous injection and assigning a score of 0, 1, 2, 3, or 4; and determining that the responses at one or more injection sites in the same subcutaneous injection in a human subject will have the same score. In one embodiment, subcutaneous administration to the animal subject comprises a single injection. In one embodiment, subcutaneous administration to the human subject comprises a single injection. In one embodiment, subcutaneous administration to the animal subject comprises two or more injections. In one embodiment, subcutaneous administration to the human subject comprises two or more injections. Attached Figure Description

[0012] Figures 1A to 1B This data includes information on the injection of 10 mL of GGL+rHuPH20 into miniature pigs using different needle sizes. Needle size selection is used to control injection time. Figure 1A It contains data obtained using a syringe pump. Figure 1B It contains data obtained using HVAI.

[0013] Figure 2 Data from human studies are provided, demonstrating that a miniature pig model can predict the timing of HVAI injections in humans. The data were obtained from a human study (Part B) in which 10 mL was injected over 30 seconds.

[0014] Figures 3A to 3B The data contained demonstrate that a miniature pig model can estimate HVAI injection site responses. Figure 3A Data on injection site responses in miniature pigs were provided. Figure 3B It provides data on human injection site responses. Detailed Implementation

[0015] Combination dosing regimens are provided, which include administration of hyaluronidase; and administration of gamma globulin to assess the clinical injection time of a 10 ml high-volume autoinjector (HVAI) containing recombinant human hyaluronidase (rHuPH20).

[0016] A. Definition

[0017] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Unless otherwise stated, all patents, patent applications, published applications and publications, and GenBank references mentioned throughout this disclosure are also referenced herein. ® Sequences, databases, websites, and other publicly available material are incorporated herein by reference in their entirety. Where a term in this document has multiple definitions, the definition provided in this section shall prevail. In the reference to URLs or other such identifiers or addresses, it should be understood that such identifiers can change, and specific information on the Internet can be rediscovered and revisited, but equivalent information can be found by searching the Internet. Such references demonstrate the availability and public dissemination of this information.

[0018] As used in this article, the term "combination dosing regimen" refers to administering at least two components together to a patient.

[0019] As used herein, the term “treatment” means, in a subject with a prior illness, alleviating a specified condition, eliminating or reducing the symptoms of the condition, slowing or eliminating the progression, invasion or spread of the condition, and reducing or delaying the recurrence of the condition.

[0020] As used in this article, the term "prevention" means stopping the development of a disease, symptom, or condition, or reducing the risk of its development or alleviating its symptoms.

[0021] As used herein, the term "injection site reaction" refers to a side effect occurring at or near the site of the infusion / injection. This includes pain or discomfort, redness, swelling, itching, bruising, lumps, infectious complications (cellulitis or abscess), and irritation.

[0022] As used herein, soluble hyaluronidase is a form of hyaluronidase that is not GPI-anchored, is soluble under physiological conditions, and is secreted upon expression. Hyaluronidases such as sheep and bovine hyaluronidase exist as soluble hyaluronidases. Human PH20 hyaluronidase does not exist as a soluble hyaluronidase. It is known in the art that removing all or part of the GPI anchor produces a soluble form.

[0023] As used herein, the term "rHuPH20" refers to a soluble hyaluronidase composition produced when a nucleic acid encoding residues 36-482 of SEQ ID NO: 1 is expressed in mammalian cells, such as CHO cells or other glycosylated cells. For expression in cells, the encoding nucleic acid is linked to a native (residues 1-35 of SEQ ID NO: 1) or heterologous signal sequence for transporting and secreting the encoded polypeptide. The resulting secretory soluble glycoprotein is a heterogeneous mixture of polypeptides, including polypeptides terminated at residues 479, 480, 481, and 482, and composed of residues 36-479, 36-480, 36-481, and 36-482 of SEQ ID NO: 1. Shorter C-terminal truncated forms may also be included.

[0024] As used in this article, “combination therapy” refers to a treatment for a single disease in which two or more therapeutic agents (such as at least two or at least three therapeutic agents) are administered to the subject.

[0025] As used herein, “hyaluronidase activity” refers to the ability of the enzyme to catalyze the cleavage of hyaluronic acid. The United States Pharmacopeia (USP) XXII assay for hyaluronidase indirectly determines hyaluronidase activity by measuring the amount of higher molecular weight hyaluronic acid or acetylated hyaluronic acid (HA) substrate remaining after allowing the enzyme to react with HA at 37°C for 30 minutes (USP XXII-NF XVII (1990) 644-645 United States Pharmacopeia Convention, Inc, Rockville, MD). Reference standard solutions can be used in the assay to determine the relative activity (in units) of any hyaluronidase. In vitro assays for determining the hyaluronidase activity of hyaluronidases such as PH20, including soluble PH20 and esPH20, are known in the art and are described herein. Exemplary assays include a microturbidity assay, which indirectly measures the cleavage of hyaluronidase on hyaluronic acid by detecting the insoluble precipitate formed when uncleaved hyaluronic acid binds to serum albumin, and a biotinylated hyaluronic acid assay, which indirectly measures the cleavage of hyaluronic acid by detecting the remaining biotinylated hyaluronic acid non-covalently bound to the wells of a microtiter plate with a streptavidin-horseradish peroxidase conjugate and a chromogenic substrate. For example, reference standards can be used to generate a standard curve to determine the unit activity of the hyaluronidase being tested.

[0026] As used herein, specific activity refers to the unit of activity per mg of protein. The milligrams of hyaluronidase are defined by the absorbance of the solution at 280 nm, assuming a molar extinction coefficient of approximately 1.7, in units of M⁻¹cm⁻¹.

[0027] As used herein, “neutral activity” refers to the ability of the PH20 peptide to enzymatically catalyze the cleavage of hyaluronic acid at a neutral pH (e.g., pH 7.0 or about).

[0028] The term “antibody” is used in the broadest sense and specifically includes monoclonal antibodies (including full-length monoclonal antibodies), multispecific antibodies (e.g., bispecific antibodies), and antibody fragments that exhibit the desired biological activity or function.

[0029] Antibodies typically consist of two heavy chains and two light chains, each comprising a variable domain and a constant domain. Each variable domain contains a hypervariable region, which in turn contains three complementarity-determining regions (CDRs), flanked by four segments of a frame region. As used herein, a “frame region” comprises all four segments FR1, FR2, FR3, and FR4 located flanking the set of three hypervariable regions.

[0030] For example, antibodies can be chimeric, humanized, or human antibodies, and can be antigen-binding fragments of these antibodies. Antibodies are typically produced by immunizing animals with antigens and can be produced through recombinant techniques or by, for example, the synthesis of amino acid sequences. An "antibody fragment" includes a portion of a full-length antibody, typically its antigen-binding region or variable region. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; biantibodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies, such as bispecific antibodies, formed, for example, from antibody fragments. A "functional fragment" essentially retains binding to the antigen of the full-length antibody and retains its biological activity.

[0031] As used herein, a “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous population of antibodies, meaning that individual antibodies in the population are identical except for the possibility of naturally occurring mutations present in small amounts. Monoclonal antibodies are highly specific, targeting a single antigenic site. Furthermore, each monoclonal antibody targets a single determinant on an antigen, unlike conventional (polyclonal) antibody formulations which typically comprise different antibodies targeting different determinants (epitopes). The modifier “monoclonal” indicates the characteristic of antibodies obtained from a substantially homogeneous population of antibodies and should not be construed as requiring the antibody to be produced by any particular method. For example, monoclonal antibodies can be prepared by a hybridoma method first described by Kohler (1975), Nature 256:495, or by a recombinant DNA method (see, for example, U.S. Patent No. 4,816,567). “Monoclonal antibodies” can also be isolated from phage antibody libraries.

[0032] As used in this article, "polyclonal antibody" refers to a mixture of immunoglobulin molecules secreted in response to a specific antigen. Polyclonal antibodies are produced by different clones of plasma B cells, resulting in a heterogeneous population of antibodies that interact with different epitopes on the same antigen.

[0033] As used herein, a “GPI anchoring signal sequence” is a C-terminal sequence of an amino acid that guides the addition of a pre-formed GPI anchor to the ER lumen of a polypeptide. GPI anchoring signal sequences are present in precursor polypeptides of GPI-anchored polypeptides, such as the GPI-anchored PH20 polypeptide. C-terminal GPI anchoring signal sequences typically contain a major hydrophobic region of 8–20 amino acids, preceded by a hydrophilic spacer region of 8–12 amino acids immediately downstream of the ω-site or GPI anchoring site. GPI anchoring signal sequences can be identified using methods well known in the art, such as, but not limited to, computer methods and algorithms (see, for example, 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 readily available on bioinformatics websites, such as the ExPASy proteomics tools site (e.g., the World Wide Web site expasy.ch / tools / ).

[0034] As used in this article, sequence identity refers to the correlation between peptides in a nucleic acid molecule. Sequence identity can be assessed by aligning two sequences and calculating the number of differences between the aligned portion and the sequence it was compared to. Whether any two molecules have at least 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% "identical" or "homologous" nucleotide or amino acid sequences can be determined using known computer algorithms (such as the "FASTA" program) with default parameters, for example, as in Pearson (1988) Proc. Natl. Acad. Sci. USA 85:2444 (other programs include the GCG package (Devereux (1984) Nucleic Acids Research 12:387), BLASTP, BLASTN, FASTA (Altschul (1990) J. Mol. Biol. 215:403; Guide to Huge Computers, Bishop, ed., Academic Press, 1994, and Carrillo (1988) SIAM J. Applied Math 48:1073). For example, the BLAST function in the National Center for Biotechnology Information database can be used to determine identity. Other commercially or publicly available programs include the DNAStar “MegAlign” program and the University of Wisconsin-Glass Group of Genetics (UWG) “Gap” program. The percentage of homology or identity between protein and / or nucleic acid molecules can be determined, for example, by comparing sequence information using the GAP computer program (e.g., Needleman (1970) J. Mol. Biol. 48:443, as revised by Smith and Waterman (1981) Adv. Appl. Math. 2:482). In short, the GAP program defines similarity as the number of similar alignment symbols (i.e., nucleotides or amino acids) divided by the total number of symbols in the shorter of the two sequences. The default parameters of the GAP procedure may include: (1) a univariate comparison matrix (containing values ​​of 1 for identity and 0 for non-identity) and a weighted comparison matrix of Gribskov (1986) Nucl. Acids Res. 14:6745, as described in Atlas of Protein Sequence and Structure, edited by Schwartz and Dayhoff, National Biomedical Research Foundation, pp. 353-358 (1979); (2) a penalty of 3.0 for each vacancy and an additional penalty of 0.10 for each symbol in each vacancy; and (3) no penalty for terminal vacancy.

[0035] Therefore, as used herein, the term “identity” or “homology” refers to a comparison between a test peptide and a reference peptide or polynucleotide.

[0036] As used herein, the term "at least 90% identical" refers to a percentage of identity of 90 to 99.99% relative to a reference nucleic acid or amino acid sequence of the polypeptide. A level of identity of 90% or higher indicates the following: assuming, for illustrative purposes, comparing 100 amino acid lengths of test and reference polypeptides, no more than 10% (i.e., 10 out of 100) of the amino acids in the test polypeptide differ from those in the reference polypeptide. A similar comparison can be made between test and reference polynucleotides. This difference can be represented as point mutations randomly distributed throughout the length of the polypeptide, or they can cluster at one or more sites of different lengths up to a maximum permissible difference, such as 10 / 100 amino acid differences (approximately 90% identity). Difference is defined as nucleic acid or amino acid substitutions, insertions, or deletions. At homology or identity levels above approximately 85%–90%, results should be independent of the procedure and the set of vacancy parameters; such high levels of identity can be readily assessed, typically by manual alignment without software dependence.

[0037] As used herein, an aligned sequence refers to a sequence that is aligned to a corresponding position in a nucleotide or amino acid sequence using homology (similarity and / or identity). Typically, an alignment is of two or more sequences that are associated with 50% or higher identity. An aligned sequence set refers to two or more sequences aligned at the corresponding positions and may include aligned sequences derived from RNA (such as ESTs and other cDNAs) that are aligned to genomic DNA sequences.

[0038] As used herein, “denaturing conditions” refers to any condition or reagent that, when exposed to a protein, typically results in the loss or partial loss of the protein’s tertiary or secondary structure, leading to degradation or denaturation. Denaturing conditions can cause effects such as loss or reduction of activity, loss or reduction of solubility, aggregation, and / or crystallization.

[0039] As used herein, “resistance to denaturing conditions” refers to any reduction or elimination of a protein’s properties or activity that are associated with or caused by denaturation. For example, denaturation is associated with or causes increased crystallization or aggregation, decreased solubility, or decreased activity. Therefore, resistance to denaturation means that, compared to a reference protein (e.g., an unmodified enzyme), a protein exhibits reduced aggregation or crystallization, increased solubility, or increased or higher activity (e.g., hyaluronidase activity) when exposed to denaturing conditions.

[0040] As used in this article, the stability of modified PH20 hyaluronidase means that it exhibits resistance to denaturation caused by denaturing conditions or denaturing agents.

[0041] To make this disclosure clear and, rather than limiting, the detailed description is divided into the following sections.

[0042] B. Soluble hyaluronidase

[0043] Soluble hyaluronidases include any hyaluronidase that is secreted from the cell at the time of expression and exists in a soluble form. Such soluble hyaluronidases include, for example, but not limited to, bacterial soluble hyaluronidases, non-human soluble hyaluronidases such as bovine PH20 and sheep PH20, human soluble PH20 and its variants. Typically, a soluble form of PH20 is produced using a protein expression system that promotes proper N-glycosylation to ensure the retention of peptide activity, as 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).

[0044] Soluble PH20 hyaluronidase is available, and for example, under the trademark ENHANZE. ® For Sale. ENHANZE ® The technology provides a drug delivery technique that utilizes soluble hyaluronidase to facilitate the delivery of injected drugs and fluids. When formulated with or administered with other drugs, ENHANZE ® The technology reduces the treatment burden on patients. It allows for large-volume subcutaneous injections while increasing the dispersion and absorption of co-administered therapies. ® The technology has been marketed along with biologics; prior to this description, it has not been used to deliver specific small molecules, nor has it been used to deliver therapeutic agents for treating and / or preventing cancer.

[0045] rHuPH20 refers to a composition produced when a nucleic acid encoding residues 36-482 of SEQ ID NO: 26 is expressed in cells such as CHO cells, typically linked to a native or heterologous signal sequence (residues 1-35 of SEQ ID NO: 26). rHuPH20 is produced by expressing nucleic acid molecules (such as those encoding amino acids 1-482 as shown in SEQ ID NO: 26) in mammalian cells. Translational processing removes the 35-amino acid signal sequence. As produced in culture media, heterogeneity exists at the C-terminus, such that the product named rHuPH20 comprises a mixture of substances that may include any or more of peptides 36-480, 36-481, and 36-482 of SEQ ID NO: 26, as well as some shorter peptides of varying abundance. rHuPH20 and soluble hyaluronidase are produced in cells that promote N-glycosylation, such as CHO cells (e.g., DG44 CHO cells). PH20 is a glycoprotein and, as is known in the art, requires glycosylation retention activity. See, for example, 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, as well as the elimination of glycosylation, on the activity of the soluble form of PH20. These patents and publications also describe and exemplify soluble C-terminal truncated forms of PH20.

[0046] 1. Soluble human pH 20 form

[0047] Soluble hyaluronidases include bovine and ovine PH20, as well as their recombinant and humanized forms. Human PH20 in nature contains a GPI anchor and is present in connection with sperm cells; it is insoluble. Its C-terminal truncated form is soluble. Soluble forms of recombinant human PH20 have been produced and can be used in the compositions, combinations, and methods described herein. Descriptions and production of such soluble forms of PH20 are described in, for example, 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, each incorporated herein by reference. Soluble hyaluronidases therefore include forms of human PH20 that are neutrally active hyaluronidases and whose activity requires glycosylation.

[0048] SEQ ID NO: 1 shows the sequence of the precursor polypeptide; the mature PH20 polypeptide (residues 36-509); the soluble forms also include those with amino acid truncation at the N-terminus, such as the deletion of the first one, two, three or four residues, such that the resulting polypeptide has an N-terminus, for example at residues 36, 37, 38, 39 or 40, and a C-terminus at residues 465 to 500, and variants thereof, including but not limited to variants discussed below, variants known in the art and allelic variants.

[0049] The hyaluronidase used in the compositions, combinations, and methods described herein is a soluble, neutral-active hyaluronidase. An example is the soluble C-terminal truncated form of mature human PH20. Soluble forms having hyaluronidase activity include, but are not limited to, those truncated at residues 465 to 500 of Sequence ID No. 1 and secreted upon expression. An example is a form having SEQ ID NO: The sequence of 1 is 36-465, 36-466, 36-467, 36-468, 36-469, 35-470, 36-471, 36-472, 36-474, 36-475, 36-476, 35-4 77, 36-478, 36-479, 36-480, 36-481, 36-482, 36-483, 35-484, 36-485, 36-486, 36-487, 36-488, 36-489 The polypeptides of 36-490, 35-491, 36-492, 36-493, 36-494, 36-495, 36-496, 36-497, 35-498, 36-499 and 36-500, and the N-terminal truncated forms of each of the above lacking two to five residues at the N-terminus, such as, for example, 37-368, 38-468, and any other form that exhibits hyaluronidase activity at neutral pH (such as pH in the range of 7.0-7.4).

[0050] Therefore, such soluble forms include truncated forms of the mature human PH20 form that lack all or part of the C-terminal GPI anchor, provided that the hyaluronidase is soluble and retains hyaluronidase activity. Soluble forms are secreted when expressed in mammalian cells and encode signal sequences such as residues 1-35 of SEQ ID NO. 1, or heterologous signal sequences that are cleaved by cells to achieve secretion. Soluble forms are those lacking the signal peptide when expressed in cells. Soluble hyaluronidases also include variants of the soluble PH20 polypeptide that exhibit hyaluronidase activity. Variants include those related to SEQ ID NO: 1 PH20 peptide 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 The polypeptides of any one of -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 have at least 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher sequence identity. Amino acid variants include conserved and non-conserved insertions, deletions, or substitutions, and include modifications, alone or in combination, such as those detailed 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, thereby providing a structure / function map of PH20; those skilled in the art can identify the replaced residues and subsequent changes in properties and activities, such as those used to achieve increased enzyme activity, increased stability under denaturing conditions, and residues that are replaced or deleted that reduce or eliminate enzyme activity.

[0051] It should be understood that residues essential to or otherwise required for hyaluronidase activity, such as any residues described above or those known to those skilled in the art, are generally unchanged and cannot be altered except for possible conserved amino acid substitutions. These include, for example, active site residues. For instance, amino acid residues 111, 113, and 176 of the human PH20 polypeptide or its soluble form (corresponding to residues in the mature PH20 polypeptide) are generally unchanged and do not change. Other residues that impart disulfide bonds necessary for glycosylation and proper folding may also remain unchanged.

[0052] Soluble human PH20 hyaluronidase is GPI-anchored and becomes soluble by removing all or part of the GPI anchor at the C-terminus. This truncation may remove all or only some of the GPI anchor attachment sequence. However, the resulting polypeptide is soluble. In cases where 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 may be retained, provided the polypeptide is soluble. Polypeptides containing one or more amino acids of the GPI anchor are referred to as extended soluble hyaluronidase. Those skilled in the art can determine whether a polypeptide is GPI-anchored using methods well known in the art. These methods include, but are not limited to, using known algorithms to predict the presence and location of the GPI anchor attachment signal sequence and ω-site, and performing solubility analysis before and after digestion with phosphatidylinositol-specific phospholipase C (PI-PLC) or D (PI-PLD).

[0053] Soluble hyaluronidases (such as those shown in SEQ ID NO: 52-57) terminating at, for example, residues 495, 496, 497, 498, 499, and 500 of SEQ ID NO: 1 can be generated by C-terminal truncation of any native GPI-anchored hyaluronidase, such that the resulting polypeptide is soluble and contains one or more amino acid residues from the GPI anchoring signal sequence (see, for example, U.S. Patent No. 8,927,249). These include hyaluronidases that are neutrally active, soluble, contain amino acid substitutions, and have at least 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, or higher sequence identity with any of SEQ ID NO: 52-57.

[0054] Typically, for use in the compositions, combinations, and methods described herein, soluble human hyaluronidases, such as soluble human PH20, and variants of PH20 having, for example, at least 91%, 95%, or 98% sequence identity with it, including those lacking 1 to 5 N-terminal residues, are used. The hyaluronidases used in the schemes, combinations, compositions, and methods described herein can be recombinantly generated or purified or partially purified from natural sources, such as testicular extracts. Methods for generating recombinant proteins, including recombinant hyaluronidases, are well known in the art.

[0055] Recombinant soluble forms of human PH20 have been produced and can be used in the compositions, combinations, and methods provided herein. For example, refer to SEQ ID NO: 1, which describes the sequence of the full-length precursor PH20, including the signal sequence (residues 1-35), and soluble forms including, but not limited to, the C-terminal truncated polypeptide of human PH20 shown in SEQ ID NO: 1, having SEQ ID NO: The C-terminal amino acid residues 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 amino acid sequence shown in 1, or a polypeptide exhibiting at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher sequence identity when compared with an unmodified sequence at soluble pH 20, is active at neutral pH, and is soluble (secreted into the culture medium when expressed in mammalian cells). The soluble form of human PH20 typically includes amino acids 36-464 as shown in SEQ ID NO: 1, terminating at residues 465-500, and optionally includes a deletion of 1-3 amino acids at the N-terminus (i.e., lacking 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. Therefore, mature soluble polypeptides include those containing 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 including 500 of SEQ ID NO: 1. Examples of soluble hyaluronidases are soluble human PH20 polypeptides of 442, 443, 444, 445, 446, or 447 amino acids in length, such as those shown above, and their variants, which have, for example, at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity and retain hyaluronidase activity.The production of such soluble forms of recombinant human PH20 is 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.

[0056] Typically, protein expression systems that promote proper N-glycosylation to ensure the retention of peptide activity are used to produce a soluble form of PH20, as glycosylation is important for the catalytic activity and stability of hyaluronidase. Such cells include, for example, Chinese hamster ovary (CHO) cells (e.g., DG44 CHO cells).

[0057] Compositions recombinantly derived from mammalian cells (such as CHO cells) are referred to as rHuPH20. It refers to the composition produced by expressing a nucleic acid encoding residues 36-482 of SEQ ID NO: 1 in cells (such as CHO cells), typically linked to a native (residues 1-35 of SEQ ID NO: 1) or heterologous signal sequence. rHuPH20 is produced by expressing nucleic acid molecules such as amino acids 1-482 (as shown in SEQ ID NO: 1) or 36-482 with a heterologous signal sequence. Post-translational processing removes the 35-amino acid signal sequence, producing a polypeptide or mixture of polypeptides, including those shown in SEQ ID NO: 3 and 44-49. Heterogeneity exists at the C-terminus, as in those produced in culture media, such that the product named rHuPH20 comprises a mixture of substances that may include any or more of SEQ ID NO: 3 and 44-49 in varying abundances. Typically, soluble hyaluronidase rHuPH20 is produced in cells that promote proper N-glycosylation to preserve activity, such as CHO cells (e.g., DG44 CHO cells). The activity of human soluble PH20 hyaluronidase requires glycosylation. When recombinantly generated from a vector encoding residues 36-582, the most abundant substance is a 446-amino acid polypeptide corresponding to residues 36-481 of SEQ ID NO: 1. The specific distribution of the resulting polypeptide may depend on the specific production method. Exemplary methods for producing high levels of PH20 are detailed, for example, in U.S. Patents 8,187,855 and 8,343,487.

[0058] 2. Glycosylation of hyaluronidase

[0059] Glycosylation (including N-linked and O-linked glycosylation) can be important for the catalytic activity and stability of some hyaluronidases, including soluble PH20 hyaluronidase. For some hyaluronidases, removal of N-linked glycosylation can lead to near-complete inactivation of hyaluronidase activity. For such hyaluronidases, the presence of N-linked glycans may be important for the production of active enzymes.

[0060] N-linked oligosaccharides are classified into several major types (oligomannoses, complexes, hybrids, and sulfateds), all of which have an amide nitrogen-linked (Man)3-GlcNAc-GlcNAc-core via an Asn residue belonging to the -Asn-Xaa-Thr / Ser- sequence (where Xaa is not Pro). Glycosylation of thrombin C at the -Asn-Xaa-Cys- site has been reported. In some cases, hyaluronidases (such as PH20 hyaluronidase) may contain both N-glycosidic bonds and O-glycosidic bonds. For example, PH20 has both O-linked and N-linked oligosaccharides. Six potential N-linked glycosylation sites are present at N82, N166, N235, N254, N368, and N393 of human PH20 illustrated in SEQ ID NO: 1.

[0061] 3. Variations

[0062] As discussed above, variants of PH20 are known to those skilled in the art or can be readily prepared based on the techniques and knowledge available in the art. Variants include those with amino acid substitutions, insertions, and deletions. Variants of soluble PH20 peptides with 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 structural / functional diagram of human PH20, detailing the role of amino acid substitutions at each residue in the catalytic domain of PH20. These patents provide approximately 7,000 examples in which the effects of substituting each amino acid with 15 other amino acids on activity and stability are identified and described. Based on these patents and earlier publications / patents, virtually all variants of soluble PH20 peptides known in the art have been described. Those skilled in the art can readily prepare soluble hyaluronidases and their variants, and are familiar with the properties of the resulting hyaluronidases.

[0063] Other variations are also known to those skilled in the art and can be used in the combinations, schemes, and methods described herein. For example, see International PCT Publications WO2020 / 022791 and WO2020197230A, which are incorporated by reference and describe modified PH20 polypeptides. These polypeptides include variations of PH20 polypeptides that typically span residues 38-468 and include substitutions, insertions, and deletions. These variations include, for example, changes of one or more amino acid residues S343E, I344N, M345T, M348K, K349E, L353A, L354I, N356E, and I361T (refer to SEQ ID NO: 1), as well as other changes including approximately 15 amino acid variations and truncation at the N-terminus and C-terminus. Variations containing such modifications and other modifications are shown in SEQ ID NO: 60-115 of International PCT Publication WO2020 / 022791. Examples of these peptides are the peptide of SEQ ID NO: 99, which, and reproduced below, is SEQ ID NO: 2. International PCT Publication WO2021 / 150079 provides variant PH20 peptides, which are described as having increased stability relative to unmodified PH20 (such as those in rHuPH20). These variant peptides have been shown to possess PH20 activity and are described as having use in co-administration with other pharmaceutical agents.

[0064] C. Application methods and procedures

[0065] 1. Application method

[0066] In one embodiment, each of hyaluronidase and gamma globulin may be administered to the patient via injection. In another embodiment, hyaluronidase and gamma globulin may be administered subcutaneously. For example, hyaluronidase and gamma globulin may be administered subcutaneously into the abdominal tissue, leg, or arm of the patient.

[0067] Compositions intended for administration to a patient via injection (e.g., subcutaneous) may also contain suitable inert additives, stabilizers, carriers, or excipients. In one embodiment, the injectable composition contains histidine. In one embodiment, the injectable composition contains sodium chloride. In one embodiment, the injectable composition contains polysorbate. In one embodiment, the polysorbate comprises polysorbate 80. In one embodiment, the injectable composition contains an antioxidant. In one embodiment, the antioxidant comprises methionine.

[0068] This article demonstrates and describes the enhanced dispersion of co-injected gamma globulin when administered in combination with hyaluronidase. Hyaluronidase temporarily promotes dispersion by reducing the viscosity of the interstitial space through depolymerization of hyaluronic acid. The permeability barrier in these tissues returns to pre-injection levels within 24 to 48 hours following hyaluronidase injection. This allows for the administration of a larger volume of gamma globulin to the patient in a single injection.

[0069] In one embodiment, gamma globulin is administered at a concentration of about 10 mg / kg or about 50 mg / kg. In another embodiment, gamma globulin is administered at a concentration of about 100 mg / kg.

[0070] 2. Plan

[0071] A regimen for administering gamma globulin in combination with soluble hyaluronidase is provided. Gamma globulin is typically formulated as a subcutaneous injection solution at an effective concentration, and hyaluronidase is provided as a composition containing an effective concentration of soluble hyaluronidase for delivering an effective amount of hyaluronidase, such as 1 mL to 5 mL or 1 mL to 3 mL.

[0072] Hyaluronidase is administered in an amount suitable for allowing the administration of approximately 100 mg of gamma globulin to a patient, according to the protocols and compositions provided herein. Exemplary ranges include, but are not limited to, amounts suitable for administering doses from 10 mg to 2000 mg to a patient or doses of at least 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600 mg, 1700 mg, 1800 mg, 1900 mg, or 2000 mg or more to a patient. In other embodiments, hyaluronidase is administered in an amount suitable to allow for the administration of at least or about 100 mg. It should be understood that a skilled practitioner can determine a specific dosage, which can depend on various parameters, including the patient's condition, age, and other factors.

[0073] In exemplary embodiments, doses from 100 mg to 2000 mg are used, such as, but not limited to, doses from 900 mg to 2000 mg, for example, doses from at least 10 mg to 1000 mg, for example, doses of at least 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 4... Gamma globulin may be administered in doses of 0.00 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600 mg, 1700 mg, 1800 mg, 1900 mg, or 2000 mg. Exemplary gamma globulin may be administered in doses from 1000 mg to 2000 mg, or in amounts between such doses. For example, gamma globulin may be administered at a dose of approximately 1500 mg, or at a dose of approximately 1750 mg, or at a dose of approximately 1900 mg, or at a dose of approximately 1950 mg.

[0074] 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 daily. 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 from about once daily to about six times daily. In one embodiment, the administration of the disclosed formulation lasts for less than about seven days. In another embodiment, the administration lasts for more than about six, ten, fourteen, twenty-eight days, two months, six months, or one year. In some cases, continuous dosing is achieved and maintained as long as necessary.

[0075] Hyaluronidase has been used clinically since the 1950s. For example, rHuPH20, approved by the FDA in 2004, has shown good tolerability in clinical evaluations at subcutaneous doses up to 96,000 U, where U is a USP unit. For example, for the purposes of this article, the dosage of hyaluronidase administered is 2,000 U to 15,000 U (such as, but not limited to, 5,000 U to 15,000 U, such as 6,000 U to 12,000 U, 8,000 U to 12,000 U) (such as doses of exactly or about 10,000 U, such as 10,000 U). The volume of hyaluronidase administered ranges from (or about) 0.5 mL to 10 mL, such as 1 mL to 5 mL, or ranges from (or about) 1 mL to 3 mL (such as 1 mL); this volume is a function of the specific activity of a particular formulation of hyaluronidase. The specific quantity depends on the parameters as understood by those skilled in the art.

[0076] In one embodiment, a combination dosing regimen is provided, comprising administering hyaluronidase; and gamma globulin, wherein the hyaluronidase is administered at a dose of 4,000 U to 15,000 U; and the gamma globulin is administered at a dose of 10 mg / kg to 100 mg / kg, wherein the combination dosing regimen is administered once every 3 months to once every 6 months.

[0077] 3. Methods for determining injection parameters

[0078] In one aspect, this disclosure provides a method for determining one or more parameters of a formulation injection in a human subject, the method comprising: The preparation containing a therapeutically effective amount of hyaluronidase was administered to animal subjects via subcutaneous injection from a large-volume autoinjector, in a dose of approximately 3 mL to approximately 50 mL. Measuring one or more parameters of subcutaneous injection in animal subjects; and The same one or more parameters for subcutaneous injection in human subjects are determined based on one or more parameters measured from animal subjects; one or more of these parameters are selected from injection time, injection force, and injection site response data.

[0079] In one embodiment, the hyaluronidase is a soluble hyaluronidase. In one embodiment, the hyaluronidase is a recombinant human hyaluronidase. In one embodiment, the hyaluronidase is a recombinant human hyaluronidase PH20 enzyme. In one embodiment, the soluble hyaluronidase comprises a sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity with the amino acid sequence described elsewhere herein. In one embodiment, the soluble hyaluronidase comprises a sequence having at least 95% amino acid sequence identity with the amino acid sequence described elsewhere herein. In one embodiment, the recombinant human hyaluronidase PH20 enzyme comprises a sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% amino acid sequence identity with the amino acid sequence of SEQ ID NO: 1. In another embodiment, the recombinant human hyaluronidase PH20 enzyme comprises a sequence having at least 95% amino acid sequence identity with the amino acid sequence of SEQ ID NO: 1.

[0080] In one embodiment, the hyaluronidase has values ​​of 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, and about 150 U / mL to about 300 kU / mL. L, with activities of 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 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 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 is a recombinant human hyaluronidase PH20 enzyme with an activity of about 120 kU / mL. In another embodiment, the hyaluronidase is a recombinant human hyaluronidase PH20 enzyme with a minimum activity of about 150 U / mL and a maximum activity of about 110,000 U / mL.

[0081] In one embodiment, the concentration of hyaluronidase in the formulation is 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, or about 100 U / mL to about 9,000 U / mL. L, 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 hyaluronidase at a concentration of about 1,500 U / mL to about 10,000 U / mL. 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.

[0082] In one embodiment, the formulation comprises hyaluronidase, which allows for the injection of large volumes of the formulation into subjects in need at high flow rates. In another embodiment, the formulation comprises a concentration and / or activity of hyaluronidase, which allows for the injection of large volumes of the formulation into subjects in need at high flow rates.

[0083] In one embodiment, the formulation further comprises a therapeutically effective amount of an active ingredient selected from small molecules, proteins, peptide fragments, biological agents, nanoparticles, antibodies, antibody fragments, and small molecule antiviral agents.

[0084] In one embodiment, the formulation further comprises a monoclonal antibody. In one embodiment, the formulation further comprises a polyclonal antibody. The monoclonal or polyclonal antibody can be any monoclonal or polyclonal antibody suitable for high-volume administration to a subject. In one embodiment, the monoclonal or polyclonal antibody can be any monoclonal or polyclonal antibody suitable for high-volume administration to a subject using a large-volume autoinjector. In one embodiment, a monoclonal or polyclonal antibody “suitable for high-volume administration” means a monoclonal or polyclonal antibody in which a therapeutically effective dose can be administered to a subject in a volume greater than about 3 mL, greater than about 4 mL, greater than about 5 mL, greater than about 6 mL, greater than about 7 mL, greater than about 8 mL, greater than about 9 mL, greater than about 10 mL, greater than about 11 mL, greater than about 12 mL, greater than about 13 mL, greater than about 14 mL, greater than about 15 mL, greater than about 16 mL, greater than about 17 mL, greater than about 18 mL, greater than about 19 mL, or greater than about 20 mL.

[0085] Exemplary monoclonal or polyclonal antibodies include, but are not limited to, trastuzumab, rituximab, pertuzumab, atezolizumab, immunoglobulin, daratumumab, egamod α, nivolumab, and olizumab. In one embodiment, the formulation comprises a combination of recombinant human hyaluronidase PH20 enzyme and trastuzumab (e.g., Herceptin SC). In one embodiment, the formulation comprises a combination of recombinant human hyaluronidase PH20 enzyme and rituximab (e.g., Mabthera SC). In one embodiment, the formulation comprises a combination of recombinant human hyaluronidase PH20 enzyme, trastuzumab, and pertuzumab (e.g., Phesgo). In one embodiment, the formulation comprises a combination of recombinant human hyaluronidase PH20 enzyme and atezolizumab (e.g., Tecentriq SC). In one embodiment, the formulation comprises a combination of recombinant human hyaluronidase PH20 enzyme and immunoglobulin (e.g., HyQvia). In one embodiment, the formulation comprises a combination of recombinant human hyaluronidase PH20 enzyme and daratumumab (e.g., Darzalex). In one embodiment, the formulation comprises a combination of recombinant human hyaluronidase PH20 enzyme and egamod α (e.g., Vyvgart). In one embodiment, the formulation comprises a combination of recombinant human hyaluronidase PH20 enzyme and nivolumab (e.g., Opdivo Qvantig). In one embodiment, the formulation comprises a combination of recombinant human hyaluronidase PH20 enzyme and ocrezumab (e.g., Ocrevus SC).

[0086] In one embodiment, the formulation further comprises gamma globulin. In another embodiment, the formulation comprises the dose of gamma globulin described elsewhere herein.

[0087] In one embodiment, the animal subject is a pig. In another embodiment, the animal subject is a miniature pig. In yet another embodiment, the animal subject is a Yucatan miniature pig.

[0088] In one embodiment, the method includes administering about 10 mL to about 20 mL of the formulation to an animal subject. In one embodiment, the method includes administering about 10 mL to about 20 mL of the formulation to a human subject. In one embodiment, about 3 mL to about 15 mL of the formulation is administered to an animal subject. In one embodiment, about 3 mL to about 15 mL of the formulation is administered to a human subject. In one embodiment, the method includes administering to an animal subject approximately 3 mL, approximately 3.1 mL, approximately 3.2 mL, approximately 3.4 mL, approximately 3.5 mL, approximately 3.6 mL, approximately 3.7 mL, approximately 3.8 mL, approximately 3.9 mL, approximately 4 mL, approximately 4.1 mL, approximately 4.2 mL, approximately 4.3 mL, approximately 4.4 mL, approximately 4.5 mL, approximately 5 mL, approximately 5.1 mL, approximately 5.2 mL, approximately 5.3 mL, approximately 5.4 mL, approximately 5.5 mL, approximately 5.6 mL, approximately 5.7 mL, approximately 5.8 mL, approximately 5.9 mL, approximately 6 mL, approximately 6.1 mL, approximately 6.2 mL, and approximately 6.3 mL. L, approximately 6.4 mL, approximately 6.5 mL, approximately 6.6 mL, approximately 6.7 mL, approximately 6.8 mL, approximately 6.9 mL, approximately 7 mL, approximately 7.1 mL, approximately 7.2 mL, approximately 7.3 mL, approximately 7.4 mL, approximately 7.5 mL, approximately 7.6 mL, approximately 7.7 mL, approximately 7.8 mL, approximately 7.9 mL, approximately 8 mL, approximately 8.1 mL, approximately 8.2 mL, approximately 8.3 mL, approximately 8.4 mL, approximately 8.5 mL, approximately 8.6 mL, approximately 8.7 mL, approximately 8.8 mL, approximately 8.9 mL, approximately 9 mL, approximately 9.1 mL, approximately 9.2 mL, approximately 9.3 mL, approximately 9.4 mL, approximately 9. 5mL, approximately 9.6mL, approximately 9.7mL, approximately 9.8mL, approximately 9.9mL, approximately 10mL, approximately 10.1mL, approximately 10.2mL, approximately 10.3mL, approximately 10.4mL, approximately 10.5mL, approximately 10.6mL, approximately 10.7mL, approximately 10.8mL, approximately 10.9mL, approximately 11mL, approximately 11.1mL, approximately 11.2mL, approximately 11.3mL, approximately 11.4mL, approximately 11.5mL, approximately 11.6mL, approximately 11.7mL, approximately 11.8mL, approximately 11.9mL, approximately 12mL, approximately 12.1mL, approximately 12.2mL, approximately 12.3mL L, approximately 12.4 mL, approximately 12.5 mL, approximately 12.6 mL, approximately 12.7 mL, approximately 12.8 mL, approximately 12.9 mL, approximately 13 mL, approximately 13.1 mL, approximately 13.2 mL, approximately 13.3 mL, approximately 13.4 mL, approximately 13.5 mL, approximately 13.6 mL, approximately 13.7 mL, approximately 13.8 mL, approximately 13.9 mL, approximately 14 mL, approximately 14.1 mL, approximately 14.2 mL, approximately 14.3 mL, approximately 14.4 mL, approximately 14.5 mL, approximately 14.6 mL, approximately 14.7 mL, approximately 14.8 mL, approximately 14.9 mL, approximately 15 mL, approximately 15.1 mL, approximately 15.2 mL, approximately 15.3 mL, approximately 15.4 mL, approximately 15.5 mL, approximately 15.6 mL, approximately 15.7 mL, approximately 15.8 mL, approximately 15.9 mL, approximately 16 mL, approximately 16.1 mL, approximately 16.2 mL, approximately 16.3 mL, approximately 16.4 mL, approximately 16.5 mL, approximately 16.6 mL, approximately 16.7 mL, approximately 16.8 mL, approximately 16.9 mL, approximately 17 mL, approximately 17.1 mL, approximately 17.2 mL, approximately 17.3 mL, approximately 17.4 mL, approximately 17.5 mL, approximately 1 7.6mL, approximately 17.7mL, approximately 17.8mL, approximately 17.9mL, approximately 18mL, approximately 18.1mL, approximately 18.2mL, approximately 18.3mL, approximately 18.4mL, approximately 18.5mL, approximately 18.6mL, approximately 18.7mL, approximately 18.8mL, approximately 18.9mL, approximately 19mL, approximately 19.1mL, approximately 19.2mL, approximately 19.3mL, approximately 19.4mL, approximately 19.5mL, approximately 19.6mL, approximately 19.7mL, approximately 19.8mL, approximately 19.9mL, approximately 20mL, approximately 2 0.1mL, approximately 20.2mL, approximately 20.3mL, approximately 20.4mL, approximately 20.5mL, approximately 20.6mL, approximately 20.7mL, approximately 20.8mL, approximately 20.9mL, approximately 21mL, approximately 21.1mL, approximately 21.2mL, approximately 21.3mL, approximately 21.4mL, approximately 21.5mL, approximately 21.6mL, approximately 21.7mL, approximately 21.8mL, approximately 21.9mL, approximately 22mL, approximately 22.1mL, approximately 22.2mL, approximately 22.3mL, approximately 22.4mL, approximately 22.5mL Approximately 22.6 mL, approximately 22.7 mL, approximately 22.8 mL, approximately 22.9 mL, approximately 23 mL, approximately 23.1 mL, approximately 23.2 mL, approximately 23.3 mL, approximately 23.4 mL, approximately 23.5 mL, approximately 23.6 mL, approximately 23.7 mL, approximately 23.8 mL, approximately 23.9 mL, approximately 24 mL, approximately 24.1 mL, approximately 24.2 mL, approximately 24.3 mL, approximately 24.4 mL, approximately 24.5 mL, approximately 24.6 mL, approximately 24.7 mL, approximately 24.8 mL, approximately 24.9 mL, or approximately 25 mL. In one embodiment, the method includes administering to a human subject approximately 3 mL, approximately 3.1 mL, approximately 3.2 mL, approximately 3.4 mL, approximately 3.5 mL, approximately 3.6 mL, approximately 3.7 mL, approximately 3.8 mL, approximately 3.9 mL, approximately 4 mL, approximately 4.1 mL, approximately 4.2 mL, approximately 4.3 mL, approximately 4.4 mL, approximately 4.5 mL, approximately 5 mL, approximately 5.1 mL, approximately 5.2 mL, approximately 5.3 mL, approximately 5.4 mL, approximately 5.5 mL, approximately 5.6 mL, approximately 5.7 mL, approximately 5.8 mL, approximately 5.9 mL, approximately 6 mL, approximately 6.1 mL, approximately 6.2 mL, approximately 6.3 mL, or approximately 6 mL.4mL, approximately 6.5mL, approximately 6.6mL, approximately 6.7mL, approximately 6.8mL, approximately 6.9mL, approximately 7mL, approximately 7.1mL, approximately 7.2mL, approximately 7.3mL, approximately 7.4mL, approximately 7.5mL, approximately 7.6mL, approximately 7.7mL, approximately 7.8mL, approximately 7.9mL, approximately 8mL, approximately 8.1mL, approximately 8.2mL, approximately 8.3mL, approximately 8.4mL, approximately 8.5mL, approximately 8.6mL, approximately 8.7mL, approximately 8.8mL, approximately 8.9mL, approximately 9mL, approximately 9.1mL, approximately 9.2mL, approximately 9.3mL, approximately 9.4mL, approximately 9.5mL, approximately 9.6mL, approximately 9.7mL, approximately 9.8mL, approximately 9.9mL, approximately 10mL Approximately 10.1 mL, approximately 10.2 mL, approximately 10.3 mL, approximately 10.4 mL, approximately 10.5 mL, approximately 10.6 mL, approximately 10.7 mL, approximately 10.8 mL, approximately 10.9 mL, approximately 11 mL, approximately 11.1 mL, approximately 11.2 mL, approximately 11.3 mL, approximately 11.4 mL, approximately 11.5 mL, approximately 11.6 mL, approximately 11.7 mL, approximately 11.8 mL, approximately 11.9 mL, approximately 12 mL, approximately 12.1 mL, approximately 12.2 mL, approximately 12.3 mL, approximately 12.4 mL, approximately 12.5 mL, approximately 12.6 mL, approximately 12.7 mL, approximately 12.8 mL, approximately 12.9 mL, approximately 13 mL, approximately 13.1 mL, approximately 13.2 mL L, approximately 13.3 mL, approximately 13.4 mL, approximately 13.5 mL, approximately 13.6 mL, approximately 13.7 mL, approximately 13.8 mL, approximately 13.9 mL, approximately 14 mL, approximately 14.1 mL, approximately 14.2 mL, approximately 14.3 mL, approximately 14.4 mL, approximately 14.5 mL, approximately 14.6 mL, approximately 14.7 mL, approximately 14.8 mL, approximately 14.9 mL, approximately 15 mL, approximately 15.1 mL, approximately 15.2 mL, approximately 15.3 mL, approximately 15.4 mL, approximately 15.5 mL, approximately 15.6 mL, approximately 15.7 mL, approximately 15.8 mL, approximately 15.9 mL, approximately 16 mL, approximately 16.1 mL, approximately 16.2 mL, approximately 16.3 mL, approximately 16.4 mL mL, approximately 16.5 mL, approximately 16.6 mL, approximately 16.7 mL, approximately 16.8 mL, approximately 16.9 mL, approximately 17 mL, approximately 17.1 mL, approximately 17.2 mL, approximately 17.3 mL, approximately 17.4 mL, approximately 17.5 mL, approximately 17.6 mL, approximately 17.7 mL, approximately 17.8 mL, approximately 17.9 mL, approximately 18 mL, approximately 18.1 mL, approximately 18.2 mL, approximately 18.3 mL, approximately 18.4 mL, approximately 18.5 mL, approximately 18.6 mL, approximately 18.7 mL, approximately 18.8 mL, approximately 18.9 mL, approximately 19 mL, approximately 19.1 mL, approximately 19.2 mL, approximately 19.3 mL, approximately 19.4 mL, approximately 19.5 mL, approximately 19.6 mL, approximately 19.7 mL, approximately 19.8 mL, approximately 19.9 mL, approximately 20 mL, approximately 20.1 mL, approximately 20.2 mL, approximately 20.3 mL, approximately 20.4 mL, approximately 20.5 mL, approximately 20.6 mL, approximately 20.7 mL, approximately 20.8 mL, approximately 20.9 mL, approximately 21 mL, approximately 21.1 mL, approximately 21.2 mL, approximately 21.3 mL, approximately 21.4 mL, approximately 21.5 mL, approximately 21.6 mL, approximately 21.7 mL, approximately 21.8 mL, approximately 21.9 mL, approximately 22 mL, approximately 22.1 mL, approximately 22.2 mL, approximately 22.3 mL The dosages are approximately 22.4 mL, 22.5 mL, 22.6 mL, 22.7 mL, 22.8 mL, 22.9 mL, 23 mL, 23.1 mL, 23.2 mL, 23.3 mL, 23.4 mL, 23.5 mL, 23.6 mL, 23.7 mL, 23.8 mL, 23.9 mL, 24 mL, 24.1 mL, 24.2 mL, 24.3 mL, 24.4 mL, 24.5 mL, 24.6 mL, 24.7 mL, 24.8 mL, 24.9 mL, or 25 mL. In one embodiment, the method includes administering the same amount of the formulation to both animal and human subjects. In one embodiment, the method includes administering approximately 10 mL of the formulation to both animal and human subjects.

[0089] In one embodiment, the formulation is in a pre-filled syringe. In one embodiment, the pre-filled syringe contains approximately 3 mL, approximately 3.1 mL, approximately 3.2 mL, approximately 3.4 mL, approximately 3.5 mL, approximately 3.6 mL, approximately 3.7 mL, approximately 3.8 mL, approximately 3.9 mL, approximately 4 mL, approximately 4.1 mL, approximately 4.2 mL, approximately 4.3 mL, approximately 4.4 mL, approximately 4.5 mL, approximately 5 mL, approximately 5.1 mL, approximately 5.2 mL, approximately 5.3 mL, approximately 5.4 mL, approximately 5.5 mL, approximately 5.6 mL, approximately 5.7 mL, approximately 5.8 mL, approximately 5.9 mL, approximately 6 mL, approximately 6.1 mL, approximately 6.2 mL, approximately 6.3 mL, approximately 6.4 mL, approximately 6.5 mL, approximately 6.6 mL, approximately 6.7 mL, and approximately 6 mL. 8mL, approximately 6.9mL, approximately 7mL, approximately 7.1mL, approximately 7.2mL, approximately 7.3mL, approximately 7.4mL, approximately 7.5mL, approximately 7.6mL, approximately 7.7mL, approximately 7.8mL, approximately 7.9mL, approximately 8mL, approximately 8.1mL, approximately 8.2mL, approximately 8.3mL, approximately 8.4mL, approximately 8.5mL, approximately 8.6mL, approximately 8.7mL, approximately 8.8mL, approximately 8.9mL, approximately 9mL, approximately 9.1mL, approximately 9.2mL, approximately 9.3mL, approximately 9.4mL, approximately 9.5mL, approximately 9.6mL, approximately 9.7mL, approximately 9.8mL, approximately 9.9mL, approximately 10mL, approximately 10.1mL, approximately 10.2mL, approximately 10.3mL Approximately 10.4 mL, approximately 10.5 mL, approximately 10.6 mL, approximately 10.7 mL, approximately 10.8 mL, approximately 10.9 mL, approximately 11 mL, approximately 11.1 mL, approximately 11.2 mL, approximately 11.3 mL, approximately 11.4 mL, approximately 11.5 mL, approximately 11.6 mL, approximately 11.7 mL, approximately 11.8 mL, approximately 11.9 mL, approximately 12 mL, approximately 12.1 mL, approximately 12.2 mL, approximately 12.3 mL, approximately 12.4 mL, approximately 12.5 mL, approximately 12.6 mL, approximately 12.7 mL, approximately 12.8 mL, approximately 12.9 mL, approximately 13 mL, approximately 13.1 mL, approximately 13.2 mL, approximately 13.3 mL, approximately 13.4 mL, approximately... 13.5mL, approximately 13.6mL, approximately 13.7mL, approximately 13.8mL, approximately 13.9mL, approximately 14mL, approximately 14.1mL, approximately 14.2mL, approximately 14.3mL, approximately 14.4mL, approximately 14.5mL, approximately 14.6mL, approximately 14.7mL, approximately 14.8mL, approximately 14.9mL, approximately 15mL, approximately 15.1mL, approximately 15.2mL, approximately 15.3mL, approximately 15.4mL, approximately 15.5mL, approximately 15.6mL, approximately 15.7mL, approximately 15.8mL, approximately 15.9mL, approximately 16mL, approximately 16.1mL, approximately 16.2mL, approximately 16.3mL, approximately 16.4mL, approximately 16.5mL, approximately 16.6 mL, approximately 16.7 mL, approximately 16.8 mL, approximately 16.9 mL, approximately 17 mL, approximately 17.1 mL, approximately 17.2 mL, approximately 17.3 mL, approximately 17.4 mL, approximately 17.5 mL, approximately 17.6 mL, approximately 17.7 mL, approximately 17.8 mL, approximately 17.9 mL, approximately 18 mL, approximately 18.1 mL, approximately 18.2 mL, approximately 18.3 mL, approximately 18.4 mL, approximately 18.5 mL, approximately 18.6 mL, approximately 18.7 mL Approximately 18.8 mL, approximately 18.9 mL, approximately 19 mL, approximately 19.1 mL, approximately 19.2 mL, approximately 19.3 mL, approximately 19.4 mL, approximately 19.5 mL, approximately 19.6 mL, approximately 19.7 mL, approximately 19.8 mL, approximately 19.9 mL, approximately 20 mL, approximately 20.1 mL, approximately 20.2 mL, approximately 20.3 mL, approximately 20.4 mL, approximately 20.5 mL, approximately 20.6 mL, approximately 20.7 mL, approximately 20.8 mL L, approximately 20.9 mL, approximately 21 mL, approximately 21.1 mL, approximately 21.2 mL, approximately 21.3 mL, approximately 21.4 mL, approximately 21.5 mL, approximately 21.6 mL, approximately 21.7 mL, approximately 21.8 mL, approximately 21.9 mL, approximately 22 mL, approximately 22.1 mL, approximately 22.2 mL, approximately 22.3 mL, approximately 22.4 mL, approximately 22.5 mL, approximately 22.6 mL, approximately 22.7 mL, approximately 22.8 mL, approximately 22.9 mL The formulation can be in the form of approximately 23 mL, 23.1 mL, 23.2 mL, 23.3 mL, 23.4 mL, 23.5 mL, 23.6 mL, 23.7 mL, 23.8 mL, 23.9 mL, 24 mL, 24.1 mL, 24.2 mL, 24.3 mL, 24.4 mL, 24.5 mL, 24.6 mL, 24.7 mL, 24.8 mL, 24.9 mL, or 25 mL. In one embodiment, the pre-filled syringe includes a needle with a gauge of approximately 20 to approximately 33. In one embodiment, the pre-filled syringe includes a BDL needle with a gauge of approximately 20 to approximately 33. In one embodiment, the pre-filled syringe includes a needle with a gauge of approximately 20 to approximately 33. In one embodiment, the pre-filled syringe includes a 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33 needle. In another embodiment, the pre-filled syringe includes a 25-gauge Terumo needle. In yet another embodiment, the pre-filled syringe includes a 25-gauge BDL needle.

[0090] In one embodiment, the method includes administering the formulation to an animal subject at a rate of about 0.08 mL / s to about 1.00 mL / s. In one embodiment, the method includes administering the formulation to a human subject at a rate of about 0.08 mL / s to about 1.00 mL / s. In one embodiment, the method includes administering the formulation to an animal subject at a rate of at least about 0.08 mL / s to about 1.0 mL / s. In one embodiment, the method includes administering the formulation to a human subject at a rate of at least about 0.08 mL / s to about 1.0 mL / s. In one embodiment, the method includes administering the formulation to an animal subject at a rate of at least or faster than about 0.08 mL / s to about 1.00 mL / s. In one embodiment, the method includes administering the formulation to a human subject at a rate of at least or faster than about 0.08 mL / s to about 1.00 mL / s. In one embodiment, the method includes administering the formulation to both the animal and human subjects at the same or about the same rate.

[0091] In one embodiment, administration of the formulation to an animal subject takes about 10 seconds to about 40 seconds. In one embodiment, administration of the formulation to a human subject takes about 10 seconds to about 40 seconds. In one embodiment, administration of the formulation to an animal subject takes at least about 10 seconds to about 40 seconds. In one embodiment, administration of the formulation to a human subject takes at least about 10 seconds to about 40 seconds. In one embodiment, administration of the formulation to an animal subject takes at least or less than about 10 seconds to about 40 seconds. In one embodiment, administration of the formulation to a human subject takes at least or less than about 10 seconds to about 40 seconds. In one embodiment, administration of the formulation to an animal subject takes about 15 seconds to about 30 seconds. In one embodiment, administration of the formulation to a human subject takes about 15 seconds to about 30 seconds. In one embodiment, administration of the formulation to an animal subject takes at least about 15 seconds to about 30 seconds. In one embodiment, administration of the formulation to a human subject takes at least about 15 seconds to about 30 seconds. In one embodiment, administration of the formulation to an animal subject takes at least or less than about 15 seconds to about 30 seconds. In one embodiment, administration of the formulation to a human subject takes at least or less than about 15 seconds to about 30 seconds.

[0092] In one embodiment, the method includes administering about 10 mL of the formulation to an animal subject at a rate of about 0.32 mL / s to about 0.42 mL / s. In one embodiment, the method includes administering about 10 mL of the formulation to a human subject at a rate of about 0.32 mL / s to about 0.42 mL / s. In one embodiment, the method includes administering about 10 mL of the formulation to both animal and human subjects at a rate of about 0.32 mL / s to about 0.42 mL / s.

[0093] Injection force

[0094] In one embodiment, one or more parameters are injection force. In one embodiment, the animal subject used for injection force measurement is a pig. In one embodiment, the animal subject used for injection force measurement is a miniature pig. In one embodiment, the animal subject used for injection force measurement is a Yucatan miniature pig.

[0095] In one embodiment, the formulation for measuring injection force further comprises gamma globulin. In another embodiment, the formulation comprises gamma globulin in the dose described elsewhere herein.

[0096] In one embodiment, the formulation for measuring injection force is administered subcutaneously to an animal subject via a large-volume autoinjector equipped with a 25-gauge needle. In another embodiment, the formulation for measuring injection force is administered subcutaneously to an animal subject via a large-volume autoinjector that dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, the large-volume autoinjector is equipped with a 25-gauge Terumo needle. In another embodiment, the large-volume autoinjector is equipped with a 25-gauge BDL needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge Terumo needle. In yet another embodiment, the pre-filled syringe is equipped with a 25-gauge BDL needle.

[0097] In one embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle. In another embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector that dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, the large-volume autoinjector is equipped with a 25-gauge Terumo needle. In another embodiment, the large-volume autoinjector is equipped with a 25-gauge BDL needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge Terumo needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge BDL needle. In yet another embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector that is the same as or substantially the same as the large-volume autoinjector used for administering the formulation to animal subjects.

[0098] In one embodiment, the method further includes determining the injection force for subcutaneous injection in a human subject by multiplying the measured injection force in an animal subject by a scaling factor. In one embodiment, the scaling factor is from about 0.80 to about 0.99. In one embodiment, the scaling factor is from about 0.85 to about 0.95. In one embodiment, the scaling factor is from about 0.88 to about 0.92.

[0099] In one implementation, the injection force is measured in Newtons (N).

[0100] In one implementation, the method includes: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.42 mL / s, wherein a large-volume autoinjector dispensed the formulation from a pre-filled syringe equipped with a 25-gauge needle. The injection force for subcutaneous injections in miniature pigs was measured to be approximately 25 N to approximately 50 N; and The injection force for the same subcutaneous injection in human subjects was determined by multiplying the injection force in miniature pigs by a scaling factor of about 0.85 to about 0.95, thus obtaining an estimated injection force of about 21 N to about 48 N.

[0101] In one implementation, the method includes: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein a large-volume autoinjector dispensed the formulation from a pre-filled syringe equipped with a 25-gauge needle. The injection force for subcutaneous injections in miniature pigs was measured to be approximately 40 N to approximately 46 N; and The injection force for the same subcutaneous injection in human subjects was determined by multiplying the injection force in miniature pigs by a scaling factor of about 0.88 to about 0.92, thus obtaining an estimated injection force of about 35 N to about 42 N.

[0102] In one embodiment, subcutaneous administration to an animal subject comprises a single injection. In one embodiment, subcutaneous administration to a human subject comprises a single injection. In one embodiment, subcutaneous administration to both animal and human subjects comprises a single injection.

[0103] In one embodiment, subcutaneous administration to an animal subject comprises two or more injections. In one embodiment, subcutaneous administration to a human subject comprises two or more injections. In one embodiment, subcutaneous administration to both animal and human subjects comprises two or more injections.

[0104] Injection time

[0105] In one embodiment, one or more measured parameters are injection time. In one embodiment, the animal subject used for injection time measurement is a pig. In one embodiment, the animal subject used for injection time measurement is a miniature pig. In one embodiment, the animal subject used for injection time measurement is a Yucatan miniature pig.

[0106] In one embodiment, the formulation for measuring injection time further comprises gamma globulin. In another embodiment, the formulation comprises the dose of gamma globulin described elsewhere herein.

[0107] In one embodiment, the formulation for measuring injection time is administered subcutaneously to an animal subject via a large-volume autoinjector equipped with a 25-gauge needle. In another embodiment, the formulation for measuring injection time is administered subcutaneously to an animal subject via a large-volume autoinjector that dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, the large-volume autoinjector is equipped with a 25-gauge Terumo needle. In another embodiment, the large-volume autoinjector is equipped with a 25-gauge BDL needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge Terumo needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge BDL needle.

[0108] In one embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle. In another embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector that dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, the large-volume autoinjector is equipped with a 25-gauge Terumo needle. In another embodiment, the large-volume autoinjector is equipped with a 25-gauge BDL needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge Terumo needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge BDL needle. In yet another embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector that is the same as or substantially the same as the large-volume autoinjector used for administering the formulation to animal subjects.

[0109] In one embodiment, the method includes determining the injection time for a subcutaneous injection in a human subject by multiplying the measured injection time of an animal subject by a scaling factor. In one embodiment, the scaling factor is from about 0.80 to about 0.99. In one embodiment, the scaling factor is from about 0.85 to about 0.95. In one embodiment, the scaling factor is from about 0.88 to about 0.92.

[0110] In one implementation, the injection time is measured in seconds.

[0111] In one implementation, the method includes: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein a large-volume autoinjector dispensed the formulation from a pre-filled syringe equipped with a 25-gauge needle. The injection time for subcutaneous injections in miniature pigs was measured to be approximately 15 seconds to approximately 35 seconds; and The injection time for the same subcutaneous injection in human subjects was determined by multiplying the injection time in miniature pigs by a scaling factor of approximately 0.85 to approximately 0.95, resulting in an estimated injection time of approximately 13 seconds to approximately 33 seconds.

[0112] In one implementation, the method includes: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein a large-volume autoinjector dispensed the formulation from a pre-filled syringe equipped with a 25-gauge needle. The injection time for subcutaneous injections in miniature pigs was measured to be approximately 18 seconds to approximately 30 seconds; and The injection time for the same subcutaneous injection in human subjects was determined by multiplying the injection time in miniature pigs by a scaling factor of approximately 0.88 to approximately 0.92, resulting in an estimated injection time of approximately 16 to approximately 28 seconds.

[0113] In one embodiment, subcutaneous administration to an animal subject comprises a single injection. In one embodiment, subcutaneous administration to a human subject comprises a single injection. In one embodiment, subcutaneous administration to both animal and human subjects comprises a single injection.

[0114] In one embodiment, subcutaneous administration to an animal subject comprises two or more injections. In one embodiment, subcutaneous administration to a human subject comprises two or more injections. In one embodiment, subcutaneous administration to both animal and human subjects comprises two or more injections.

[0115] Injection site reaction data

[0116] In one embodiment, one or more measured parameters are injection site reaction data. In one embodiment, the injection site reaction data includes one or more of the following: backleakage at the injection site, swelling volume at the injection site, swelling height at the injection site, wheal size at the injection site, wheal induration at the injection site, and wheal regression at the injection site.

[0117] In one embodiment, the animal subjects used for measuring injection site reaction data are pigs. In another embodiment, the animal subjects used for measuring injection site reaction data are miniature pigs. In yet another embodiment, the animal subjects used for measuring injection site reaction data are Yucatan miniature pigs.

[0118] In one embodiment, the formulation used for measuring injection site reaction data further comprises gamma globulin. In one embodiment, the formulation comprises gamma globulin in the dose described elsewhere herein.

[0119] In one embodiment, the formulation for measuring injection site response data is administered subcutaneously to an animal subject via a large-volume autoinjector equipped with a 25-gauge needle. In another embodiment, the formulation for measuring injection site response data is administered subcutaneously to an animal subject via a large-volume autoinjector that dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, the large-volume autoinjector is equipped with a 25-gauge Terumo needle. In another embodiment, the large-volume autoinjector is equipped with a 25-gauge BDL needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge Terumo needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge BDL needle.

[0120] In one embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle. In another embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector that dispenses the formulation from a pre-filled syringe equipped with a 25-gauge needle. In one embodiment, the large-volume autoinjector is equipped with a 25-gauge Terumo needle. In another embodiment, the large-volume autoinjector is equipped with a 25-gauge BDL needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge Terumo needle. In another embodiment, the pre-filled syringe is equipped with a 25-gauge BDL needle. In yet another embodiment, subcutaneous administration to a human subject is performed via a large-volume autoinjector that is the same as or substantially the same as the large-volume autoinjector used for administering the formulation to animal subjects.

[0121] In one implementation, the method includes: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.42 mL / s, wherein a large-volume autoinjector dispensed the formulation from a pre-filled syringe equipped with a 25-gauge needle. Measure the response at one or more injection sites in miniature pigs after subcutaneous injection and assign a score of 0, 1, 2, 3, or 4; and It is determined that responses at one or more injection sites for the same subcutaneous injection in human subjects will have the same score.

[0122] In one implementation, the method includes: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein a large-volume autoinjector dispensed the formulation from a pre-filled syringe equipped with a 25-gauge needle. Measure the response at one or more injection sites in miniature pigs after subcutaneous injection and assign a score of 0, 1, 2, 3, or 4; and It is determined that responses at one or more injection sites for the same subcutaneous injection in human subjects will have the same score.

[0123] In one embodiment, subcutaneous administration to an animal subject comprises a single injection. In one embodiment, subcutaneous administration to a human subject comprises a single injection. In one embodiment, subcutaneous administration to both animal and human subjects comprises a single injection.

[0124] In one embodiment, subcutaneous administration to an animal subject comprises two or more injections. In one embodiment, subcutaneous administration to a human subject comprises two or more injections. In one embodiment, subcutaneous administration to both animal and human subjects comprises two or more injections.

[0125] Terms of this disclosure

[0126] Clause 1. A method for determining one or more parameters of a formulation injection in a human subject, the method comprising: The preparation containing a therapeutically effective amount of hyaluronidase was administered to animal subjects via subcutaneous injection from a large-volume autoinjector, in a dose of approximately 3 mL to approximately 50 mL. Measuring one or more parameters of the subcutaneous injection in the animal subjects; and The same one or more parameters for the subcutaneous injection in the human subjects are determined based on one or more measured parameters from the animal subjects; wherein the one or more parameters are selected from injection time, injection force, and injection site response data.

[0127] Clause 2. The hyaluronidase described in accordance with Clause 1 is a soluble hyaluronidase.

[0128] Clause 3. The method according to Clause 1 or 2, wherein the hyaluronidase is recombinant human hyaluronidase.

[0129] Clause 4. The method according to any one of Clauses 1 to 3, wherein the hyaluronidase is recombinant human hyaluronidase PH20.

[0130] Clause 5. The method according to Clause 4, wherein the recombinant human hyaluronidase PH20 enzyme comprises a sequence having at least 95% amino acid sequence identity with the amino acid sequence of SEQ ID NO: 1.

[0131] Clause 6. The method according to any one of Clauses 1 to 5, wherein the formulation further comprises a therapeutically effective amount of an active ingredient selected from small molecules, proteins, peptide fragments, biological agents, nanoparticles, antibodies, antibody fragments, and small molecule antiviral agents.

[0132] Clause 7. The method described in Clause 6, wherein the antibody comprises a monoclonal antibody or a polyclonal antibody.

[0133] Clause 8. The method according to Clause 7, wherein the monoclonal antibody or the polyclonal antibody comprises trastuzumab, rituximab, pertuzumab, atezolizumab, immunoglobulin, daratumumab, egamod α, nivolumab, or ozoglucomannan.

[0134] Clause 9. The method according to Clause 6, wherein the protein comprises γ-globulin.

[0135] Clause 10. The method according to any one of Clauses 1 to 9, wherein the animal subject is a miniature pig.

[0136] Clause 11. The method according to any one of Clauses 1 to 10, wherein the animal subject is a Yucatan miniature pig.

[0137] Clause 12. The method according to any one of Clauses 1 to 11, the method comprising administering about 10 mL to about 20 mL of the preparation to the animal subject.

[0138] Clause 13. The method according to any one of Clauses 1 to 9, wherein the method comprises administering about 10 mL to about 20 mL of the preparation to the human subject.

[0139] Clause 14. The method according to any one of Clauses 1 to 12, wherein the method comprises administering about 3 mL to about 15 mL of the preparation to the animal subject.

[0140] Clause 15. The method according to any one of Clauses 1 to 9 or 13, the method comprising administering about 3 mL to about 15 mL of the preparation to the human subject.

[0141] Clause 16. The method according to any one of Clauses 1 to 12 or 14, said method comprising administering to the animal 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, or about 6.5 mL. Approximately 6.6 mL, approximately 6.7 mL, approximately 6.8 mL, approximately 6.9 mL, approximately 7 mL, approximately 7.1 mL, approximately 7.2 mL, approximately 7.3 mL, approximately 7.4 mL, approximately 7.5 mL, approximately 7.6 mL, approximately 7.7 mL, approximately 7.8 mL, approximately 7.9 mL, approximately 8 mL, approximately 8.1 mL, approximately 8.2 mL, approximately 8.3 mL, approximately 8.4 mL, approximately 8.5 mL, approximately 8.6 mL, approximately 8.7 mL, approximately 8.8 mL, approximately 8.9 mL, approximately 9 mL, approximately 9 mL, approximately 9.1 mL, approximately 9.2 mL, approximately 9.3 mL, approximately 9.4 mL, approximately 9.5 mL, approximately 9.6 mL, approximately 9.7 mL, approximately 9.8 mL, approximately 9.9 mL, approximately 10 mL, approximately 10.1 mL, approximately 10 0.2mL, approximately 10.3mL, approximately 10.4mL, approximately 10.5mL, approximately 10.6mL, approximately 10.7mL, approximately 10.8mL, approximately 10.9mL, approximately 11mL, approximately 11.1mL, approximately 11.2mL, approximately 11.3mL, approximately 11.4mL, approximately 11.5mL, approximately 11.6mL, approximately 11.7mL, approximately 11.8mL, approximately 11.9mL, approximately 12mL, approximately 12.1mL, approximately 12.2mL, approximately 12.3mL, approximately 12.4mL, approximately 12.5mL, approximately 12.6mL, approximately 12.7mL, approximately 12.8mL, approximately 12.9mL, approximately 13mL, approximately 13.1mL, approximately 13.2mL, approximately 13.3mL, approximately 13... 0.4mL, approximately 13.5mL, approximately 13.6mL, approximately 13.7mL, approximately 13.8mL, approximately 13.9mL, approximately 14mL, approximately 14.1mL, approximately 14.2mL, approximately 14.3mL, approximately 14.4mL, approximately 14.5mL, approximately 14.6mL, approximately 14.7mL, approximately 14.8mL, approximately 14.9mL, approximately 15mL, approximately 15.1mL, approximately 15.2mL, approximately 15.3mL, approximately 15.4mL, approximately 15.5mL, approximately 15.6mL, approximately 15.7mL, approximately 15.8mL, approximately 15.9mL, approximately 16mL, approximately 16.1mL, approximately 16.2mL, approximately 16.3mL, approximately 16.4mL, approximately 16.5mL, approximately 16.6mL, approximately 16.7mL, approximately 16.8mL, approximately 16.9mL, approximately 17mL, approximately 17.1mL, approximately 17.2mL, approximately 17.3mL, approximately 17.4mL, approximately 17.5mL, approximately 17.6mL, approximately 17.7mL, approximately 17.8mL, approximately 17.9mL, approximately 18mL, approximately 18.1mL, approximately 18.2mL, approximately 18.3mL, approximately 18.4mL, approximately 18.5mL, approximately 18.6mL, approximately 18. 7mL, approximately 18.8mL, approximately 18.9mL, approximately 19mL, approximately 19.1mL, approximately 19.2mL, approximately 19.3mL, approximately 19.4mL, approximately 19.5mL, approximately 19.6mL, approximately 19.7mL, approximately 19.8mL, approximately 19.9mL, approximately 20mL, approximately 20.1mL, approximately 20.2mL, approximately 20.3mL, approximately 20.4mL, approximately 20.5mL, approximately 20.6mL, approximately 20.7mL, approximately 20.8mL mL, approximately 20.9 mL, approximately 21 mL, approximately 21.1 mL, approximately 21.2 mL, approximately 21.3 mL, approximately 21.4 mL, approximately 21.5 mL, approximately 21.6 mL, approximately 21.7 mL, approximately 21.8 mL, approximately 21.9 mL, approximately 22 mL, approximately 22.1 mL, approximately 22.2 mL, approximately 22.3 mL, approximately 22.4 mL, approximately 22.5 mL, approximately 22.6 mL, approximately 22.7 mL, approximately 22.8 mL, approximately 22.9 mL Approximately 23 mL, 23.1 mL, 23.2 mL, 23.3 mL, 23.4 mL, 23.5 mL, 23.6 mL, 23.7 mL, 23.8 mL, 23.9 mL, 24 mL, 24.1 mL, 24.2 mL, 24.3 mL, 24.4 mL, 24.5 mL, 24.6 mL, 24.7 mL, 24.8 mL, 24.9 mL, or 25 mL.

[0142] Clause 17. The method according to any one of Clauses 1 to 9, 13, or 15, said method comprising administering to the human 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, or about 6 mL. 0.5mL, approximately 6.6mL, approximately 6.7mL, approximately 6.8mL, approximately 6.9mL, approximately 7mL, approximately 7.1mL, approximately 7.2mL, approximately 7.3mL, approximately 7.4mL, approximately 7.5mL, approximately 7.6mL, approximately 7.7mL, approximately 7.8mL, approximately 7.9mL, approximately 8mL, approximately 8.1mL, approximately 8.2mL, approximately 8.3mL, approximately 8.4mL, approximately 8.5mL, approximately 8.6mL, approximately 8.7mL, approximately 8.8mL, approximately 8.9mL, approximately 9mL, approximately 9mL, approximately 9.1mL, approximately 9.2mL, approximately 9.3mL, approximately 9.4mL, approximately 9.5mL, approximately 9.6mL, approximately 9.7mL, approximately 9.8mL, approximately 9.9mL, approximately 10mL, approximately 10.1mL L, approximately 10.2 mL, approximately 10.3 mL, approximately 10.4 mL, approximately 10.5 mL, approximately 10.6 mL, approximately 10.7 mL, approximately 10.8 mL, approximately 10.9 mL, approximately 11 mL, approximately 11.1 mL, approximately 11.2 mL, approximately 11.3 mL, approximately 11.4 mL, approximately 11.5 mL, approximately 11.6 mL, approximately 11.7 mL, approximately 11.8 mL, approximately 11.9 mL, approximately 12 mL, approximately 12.1 mL, approximately 12.2 mL, approximately 12.3 mL, approximately 12.4 mL, approximately 12.5 mL, approximately 12.6 mL, approximately 12.7 mL, approximately 12.8 mL, approximately 12.9 mL, approximately 13 mL, approximately 13.1 mL, approximately 13.2 mL, approximately 13.3 mL mL, approximately 13.4 mL, approximately 13.5 mL, approximately 13.6 mL, approximately 13.7 mL, approximately 13.8 mL, approximately 13.9 mL, approximately 14 mL, approximately 14.1 mL, approximately 14.2 mL, approximately 14.3 mL, approximately 14.4 mL, approximately 14.5 mL, approximately 14.6 mL, approximately 14.7 mL, approximately 14.8 mL, approximately 14.9 mL, approximately 15 mL, approximately 15.1 mL, approximately 15.2 mL, approximately 15.3 mL, approximately 15.4 mL, approximately 15.5 mL, approximately 15.6 mL, approximately 15.7 mL, approximately 15.8 mL, approximately 15.9 mL, approximately 16 mL, approximately 16.1 mL, approximately 16.2 mL, approximately 16.3 mL, approximately 16.4 mL, approximately 16.5mL, approximately 16.6mL, approximately 16.7mL, approximately 16.8mL, approximately 16.9mL, approximately 17mL, approximately 17.1mL, approximately 17.2mL, approximately 17.3mL, approximately 17.4mL, approximately 17.5mL, approximately 17.6mL, approximately 17.7mL, approximately 17.8mL, approximately 17.9mL, approximately 18mL, approximately 18.1mL, approximately 18.2mL, approximately 18.3mL, approximately 18.4mL, approximately 18.5mL, approximately 18.6mL L, approximately 18.7 mL, approximately 18.8 mL, approximately 18.9 mL, approximately 19 mL, approximately 19.1 mL, approximately 19.2 mL, approximately 19.3 mL, approximately 19.4 mL, approximately 19.5 mL, approximately 19.6 mL, approximately 19.7 mL, approximately 19.8 mL, approximately 19.9 mL, approximately 20 mL, approximately 20.1 mL, approximately 20.2 mL, approximately 20.3 mL, approximately 20.4 mL, approximately 20.5 mL, approximately 20.6 mL, approximately 20.7 mL, approximately 20.8 mL, approximately 20.9 mL, approximately 21 mL, approximately 21.1 mL, approximately 21.2 mL, approximately 21.3 mL, approximately 21.4 mL, approximately 21.5 mL, approximately 21.6 mL, approximately 21.7 mL, approximately 21.8 mL, approximately 21.9 mL, approximately 22 mL, approximately 22.1 mL, approximately 22.2 mL, approximately 22.3 mL, approximately 22.4 mL, approximately 22.5 mL, approximately 22.6 mL, approximately 22.7 mL, approximately 22.8 mL, approximately 22. 9 mL, approximately 23 mL, approximately 23.1 mL, approximately 23.2 mL, approximately 23.3 mL, approximately 23.4 mL, approximately 23.5 mL, approximately 23.6 mL, approximately 23.7 mL, approximately 23.8 mL, approximately 23.9 mL, approximately 24 mL, approximately 24.1 mL, approximately 24.2 mL, approximately 24.3 mL, approximately 24.4 mL, approximately 24.5 mL, approximately 24.6 mL, approximately 24.7 mL, approximately 24.8 mL, approximately 24.9 mL, or approximately 25 mL.

[0143] Clause 18. The method according to any one of Clauses 1 to 17, wherein the formulation is in a pre-filled syringe.

[0144] Clause 19. The method according to Clause 18, wherein the pre-filled 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, about 6.4 mL, about 6.5 mL, about 6.6 mL, about 6.7 mL. mL, approximately 6.8 mL, approximately 6.9 mL, approximately 7 mL, approximately 7.1 mL, approximately 7.2 mL, approximately 7.3 mL, approximately 7.4 mL, approximately 7.5 mL, approximately 7.6 mL, approximately 7.7 mL, approximately 7.8 mL, approximately 7.9 mL, approximately 8 mL, approximately 8.1 mL, approximately 8.2 mL, approximately 8.3 mL, approximately 8.4 mL, approximately 8.5 mL, approximately 8.6 mL, approximately 8.7 mL, approximately 8.8 mL, approximately 8.9 mL, approximately 9 mL, approximately 9.1 mL, approximately 9.2 mL, approximately 9.3 mL, approximately 9.4 mL, approximately 9.5 mL, approximately 9.6 mL, approximately 9.7 mL, approximately 9.8 mL, approximately 9.9 mL, approximately 10 mL, approximately 10.1 mL, approximately 10.2 mL, approximately 10.3 mL L, approximately 10.4 mL, approximately 10.5 mL, approximately 10.6 mL, approximately 10.7 mL, approximately 10.8 mL, approximately 10.9 mL, approximately 11 mL, approximately 11.1 mL, approximately 11.2 mL, approximately 11.3 mL, approximately 11.4 mL, approximately 11.5 mL, approximately 11.6 mL, approximately 11.7 mL, approximately 11.8 mL, approximately 11.9 mL, approximately 12 mL, approximately 12.1 mL, approximately 12.2 mL, approximately 12.3 mL, approximately 12.4 mL, approximately 12.5 mL, approximately 12.6 mL, approximately 12.7 mL, approximately 12.8 mL, approximately 12.9 mL, approximately 13 mL, approximately 13.1 mL, approximately 13.2 mL, approximately 13.3 mL, approximately 13.4 mL, approximately 13.5 mL mL, approximately 13.6 mL, approximately 13.7 mL, approximately 13.8 mL, approximately 13.9 mL, approximately 14 mL, approximately 14.1 mL, approximately 14.2 mL, approximately 14.3 mL, approximately 14.4 mL, approximately 14.5 mL, approximately 14.6 mL, approximately 14.7 mL, approximately 14.8 mL, approximately 14.9 mL, approximately 15 mL, approximately 15.1 mL, approximately 15.2 mL, approximately 15.3 mL, approximately 15.4 mL, approximately 15.5 mL, approximately 15.6 mL, approximately 15.7 mL, approximately 15.8 mL, approximately 15.9 mL, approximately 16 mL, approximately 16.1 mL, approximately 16.2 mL, approximately 16.3 mL, approximately 16.4 mL, approximately 16.5 mL, approximately 16.6 mL, approximately 16.7mL, approximately 16.8mL, approximately 16.9mL, approximately 17mL, approximately 17.1mL, approximately 17.2mL, approximately 17.3mL, approximately 17.4mL, approximately 17.5mL, approximately 17.6mL, approximately 17.7mL, approximately 17.8mL, approximately 17.9mL, approximately 18mL, approximately 18.1mL, approximately 18.2mL, approximately 18.3mL, approximately 18.4mL, approximately 18.5mL, approximately 18.6mL, approximately 18.7mL, approximately 18. 8mL, approximately 18.9mL, approximately 19mL, approximately 19.1mL, approximately 19.2mL, approximately 19.3mL, approximately 19.4mL, approximately 19.5mL, approximately 19.6mL, approximately 19.7mL, approximately 19.8mL, approximately 19.9mL, approximately 20mL, approximately 20.1mL, approximately 20.2mL, approximately 20.3mL, approximately 20.4mL, approximately 20.5mL, approximately 20.6mL, approximately 20.7mL, approximately 20.8mL, approximately 20. 9mL, approximately 21mL, approximately 21.1mL, approximately 21.2mL, approximately 21.3mL, approximately 21.4mL, approximately 21.5mL, approximately 21.6mL, approximately 21.7mL, approximately 21.8mL, approximately 21.9mL, approximately 22mL, approximately 22.1mL, approximately 22.2mL, approximately 22.3mL, approximately 22.4mL, approximately 22.5mL, approximately 22.6mL, approximately 22.7mL, approximately 22.8mL, approximately 22.9mL, approximately 23mL The preparation may be expressed in L, approximately 23.1 mL, approximately 23.2 mL, approximately 23.3 mL, approximately 23.4 mL, approximately 23.5 mL, approximately 23.6 mL, approximately 23.7 mL, approximately 23.8 mL, approximately 23.9 mL, approximately 24 mL, approximately 24.1 mL, approximately 24.2 mL, approximately 24.3 mL, approximately 24.4 mL, approximately 24.5 mL, approximately 24.6 mL, approximately 24.7 mL, approximately 24.8 mL, approximately 24.9 mL, or approximately 25 mL.

[0145] Clause 20. The method according to Clause 18 or 19, wherein the prefilled syringe comprises a needle of about 20 to about 33.

[0146] Clause 21. The method according to any one of Clauses 18 to 20, wherein the pre-filled 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.

[0147] Clause 22. The method according to any one of Clauses 1 to 12, 14 or 16, the method comprising administering the formulation to the animal subject at a rate of about 0.08 mL / s to about 1.00 mL / s.

[0148] Clause 23. The method according to any one of Clauses 1 to 9, 13, 15 or 17, the method comprising administering the formulation to the human subject at a rate of about 0.08 mL / s to about 1.00 mL / s.

[0149] Clause 24. The method according to any one of Clauses 1 to 12, 14, 16 or 22, said method comprising administering the formulation to the animal subject at a rate of at least about 0.08 mL / s to about 1.0 mL / s.

[0150] Clause 25. The method according to any one of Clauses 1 to 9, 13, 15, 17 or 23, said method comprising administering the formulation to the human subject at a rate of at least about 0.08 mL / s to about 1.0 mL / s.

[0151] Clause 26. The method according to any one of Clauses 1 to 12, 14 or 16, the method comprising administering the formulation to the animal subject at a rate of at least or faster than about 0.08 mL / s to about 1.00 mL / s.

[0152] Clause 27. The method according to any one of Clauses 1 to 9, 13, 15 or 17, the method comprising administering the formulation to the human subject at a rate of at least or faster than about 0.08 mL / s to about 1.00 mL / s.

[0153] Clause 28. The method according to any one of Clauses 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes about 10 seconds to about 40 seconds.

[0154] Clause 29. The method according to any one of Clauses 1 to 9, 13, 15, 17, 23 or 25, wherein the administration of the formulation to the human subject takes about 10 seconds to about 40 seconds.

[0155] Clause 30. The method according to any one of Clauses 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes at least about 10 seconds to about 40 seconds.

[0156] Clause 31. The method according to any one of Clauses 1 to 9, 13, 15, 17, 23 or 25, wherein the administration of the formulation to the human subject takes at least about 10 seconds to about 40 seconds.

[0157] Clause 32. The method according to any one of Clauses 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes at least or less than about 10 seconds to about 40 seconds.

[0158] Clause 33. The method according to any one of Clauses 1 to 9, 13, 15, 17, 23 or 25, wherein the administration of the formulation to the human subject takes at least or less than about 10 seconds to about 40 seconds.

[0159] Clause 34. The method according to any one of Clauses 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes about 15 seconds to about 30 seconds.

[0160] Clause 35. The method according to any one of Clauses 1 to 9, 13, 15, 17, 23 or 25, wherein the administration of the formulation to the human subject takes about 15 seconds to about 30 seconds.

[0161] Clause 36. The method according to any one of Clauses 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes at least about 15 seconds to about 30 seconds.

[0162] Clause 37. The method according to any one of Clauses 1 to 9, 13, 15, 17, 23 or 25, wherein the administration of the formulation to the human subject takes at least about 15 seconds to about 30 seconds.

[0163] Clause 38. The method according to any one of Clauses 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes at least or less than about 15 seconds to about 30 seconds.

[0164] Clause 39. The method according to any one of Clauses 1 to 9, 13, 15, 17, 23 or 25, wherein the administration of the formulation to the human subject takes at least or less than about 15 seconds to about 30 seconds.

[0165] Clause 40. The method according to any one of Clauses 1 to 12, 14, 16, 22 or 24, said method comprising administering about 10 mL of said formulation to the animal subject at a rate of about 0.32 mL / s to about 0.42 mL / s.

[0166] Clause 41. The method according to any one of Clauses 1 to 9, 13, 15, 17, 23 or 25, said method comprising administering about 10 mL of said formulation to the human subject at a rate of about 0.32 mL / s to about 0.42 mL / s.

[0167] Clause 42. The method according to any one of Clauses 1 to 41, wherein the parameter is injection force.

[0168] Clause 43. The method according to Clause 42, wherein the animal subject is a miniature pig.

[0169] Clause 44. The method according to Clause 42 or 43, wherein the animal subject is a Yucatan miniature pig.

[0170] Clause 45. The method according to any one of Clauses 42 to 44, wherein the formulation further comprises γ-globulin.

[0171] Clause 46. The method according to any one of Clauses 42 to 45, wherein the subcutaneous administration to the animal subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

[0172] Clause 47. The method according to Clause 42 or 45, wherein the subcutaneous administration to the human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

[0173] Clause 48. The method according to any one of Clauses 42 to 47, the method comprising determining the injection force of the subcutaneous injection in the human subject by multiplying the measured injection force in the animal subject by a scaling factor.

[0174] Clause 49. The method according to Clause 48, wherein the scaling factor is about 0.80 to about 0.99.

[0175] Clause 50. The method according to Clause 48 or 49, wherein the scaling factor is about 0.85 to about 0.95; alternatively about 0.88 to about 0.92.

[0176] Clause 51. The method according to any one of Clauses 42 to 50, wherein the injection force is measured in Newtons (N).

[0177] Clause 52. The method according to any one of Clauses 42 to 51, said method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.42 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. The injection force of the subcutaneous injection in the miniature pig was measured to be approximately 25 N to approximately 50 N; and The injection force for the same subcutaneous injection in the human subjects was determined by multiplying the injection force in the miniature pigs by a scaling factor of about 0.85 to about 0.95, thereby obtaining an estimated injection force of about 21 N to about 48 N.

[0178] Clause 53. The method according to any one of Clauses 42 to 52, said method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. The injection force of the subcutaneous injection in the miniature pig was measured to be approximately 40 N to approximately 46 N; and An estimated injection force of about 35 N to about 42 N was obtained by multiplying the injection force in the miniature pigs by a scaling factor of about 0.88 to about 0.92 to determine the same subcutaneous injection force in the human subjects.

[0179] Clause 54. The method according to any one of Clauses 1 to 41, wherein the parameter is the injection time.

[0180] Clause 55. The method according to Clause 54, wherein the animal subject is a miniature pig.

[0181] Clause 56. The method described in accordance with Clause 54 or 55, wherein the animal subject is a Yucatan miniature pig.

[0182] Clause 57. The method according to any one of Clauses 54 to 56, wherein the formulation further comprises γ-globulin.

[0183] Clause 58. The method according to any one of Clauses 54 to 57, wherein the subcutaneous administration to the animal subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

[0184] Clause 59. The method according to Clause 54 or 57, wherein the subcutaneous administration to the human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

[0185] Clause 60. The method according to any one of Clauses 54 to 59, the method comprising determining the injection time of the subcutaneous injection in the human subject by multiplying the measured injection time of the animal subject by a scaling factor.

[0186] Clause 61. The method according to Clause 60, wherein the scaling factor is about 0.80 to about 0.99.

[0187] Clause 62. The method according to Clause 60 or 61, wherein the scaling factor is about 0.85 to about 0.95; optionally about 0.88 to about 0.92.

[0188] Clause 63. The method according to any one of Clauses 54 to 62, wherein the injection time is measured in seconds.

[0189] Clause 64. The method according to any one of Clauses 54 to 63, said method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. The injection time for the subcutaneous injection in the miniature pigs was measured to be approximately 15 seconds to approximately 35 seconds; and The injection time for the same subcutaneous injection in the human subjects was determined by multiplying the injection time in the miniature pigs by a scaling factor of about 0.85 to about 0.95, thereby obtaining an estimated injection time of about 13 seconds to about 33 seconds.

[0190] Clause 65. The method according to any one of Clauses 54 to 64, said method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. The injection time for the subcutaneous injection in the miniature pigs was measured to be approximately 18 seconds to approximately 30 seconds; and The injection time for the same subcutaneous injection in the human subjects was determined by multiplying the injection time in the miniature pigs by a scaling factor of about 0.88 to about 0.92, thereby obtaining an estimated injection time of about 16 seconds to about 28 seconds.

[0191] Clause 66. The method according to any one of Clauses 1 to 41, wherein the parameter is injection site reaction data.

[0192] Clause 67. The method according to Clause 66, wherein the injection site reaction data includes one or more of the following: backflow at the injection site, swelling volume at the injection site, swelling height at the injection site, swelling size of the wheal at the injection site, induration of the wheal at the injection site, and wheal regression at the injection site.

[0193] Clause 68. The method according to Clause 66 or 67, wherein the animal subject is a miniature pig.

[0194] Clause 69. The method according to any one of Clauses 66 to 68, wherein the animal subject is a Yucatan miniature pig.

[0195] Clause 70. The method according to any one of Clauses 66 to 69, wherein the formulation further comprises γ-globulin.

[0196] Clause 71. The method according to any one of Clauses 66 to 70, wherein the subcutaneous administration to the animal subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

[0197] Clause 72. The method according to any one of Clauses 66, 67 or 70, wherein the subcutaneous administration to the human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

[0198] Clause 73. The method according to any one of Clauses 66 to 72, said method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.42 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. Measure the response at one or more injection sites of the subcutaneous injection in the miniature pigs and assign a score of 0, 1, 2, 3, or 4; and It is determined that responses at one or more injection sites for the same subcutaneous injection in the human subjects will have the same score.

[0199] Clause 74. The method according to any one of Clauses 66 to 73, said method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. Measure the response at one or more injection sites of the subcutaneous injection in the miniature pigs and assign a score of 0, 1, 2, 3, or 4; and It is determined that responses at one or more injection sites for the same subcutaneous injection in the human subjects will have the same score.

[0200] Clause 75. The method according to any one of Clauses 66 to 71, 73 or 74, wherein the subcutaneous administration to the animal subject comprises a single injection.

[0201] Clause 76. The method according to any one of Clauses 66, 67, 70 or 72 to 74, wherein the subcutaneous administration to the human subject comprises a single injection.

[0202] Clause 77. The method according to any one of Clauses 66 to 71, 73 or 75, wherein the subcutaneous administration to the animal subject comprises two or more injections.

[0203] Clause 78. The method according to any one of Clauses 66, 67, 69 or 71 to 73, wherein the subcutaneous administration to the human subject comprises two or more injections.

[0204] Example

[0205] The following embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0206] Example 1: Miniature pigs as a preclinical model to predict the presence of recombinant human hyaluronidase (rHuPH20) Clinical injection time of a 10mL high-volume automated injector (HVAI)

[0207] Autoinjectors have now evolved to accommodate volumes up to 10 mL; however, a limitation of rapid, high-volume fluid flow is the presence of hyaluronic acid (HA) in the SC space. rHuPH20 depolymerizes HA to instantaneously reduce this barrier and tissue back pressure, enabling high-volume, rapid SC injection. We have developed a clinically usable 10 mL HVAI and evaluated its use in an animal model where rHuPH20 can predictively translate preclinical findings into human injection time outcomes.

[0208] 10 mL of GGL with rHuPH20 was injected into miniature pigs over 30 seconds using a pump, syringe, and 25G needle (BD or Terumo). For the BD and Terumo needles, the injection force measured by the force sensor was 67.4 N and 42.6 N, respectively. The procedure was then repeated in miniature pigs using the same two needles with an HVAI to determine mean injection times of 30.0 seconds and 19.8 seconds, respectively. Clinical trials were initiated using the same pump, syringe, and Terumo 25G needle. For the same 10 mL, 30-second SC injection, the injection force was measured to be 38.7 N. Based on these data, the predicted injection time for the human HVAI was calculated to be 27.2 seconds. The clinical trial yielded an injection time of 27.9 seconds.

[0209] Measure and record the duration of each injection. Additional endpoints include measurement of post-injection backleakage, local injection site measurements (wheal area and volume), qualitative scoring of local tissue erythema, swelling size, and induration, and standardized imaging to quantify post-injection changes in the skin.

[0210] All device assembly was performed under aseptic conditions with sterile filling and sealing caps. Test solutions were loaded into the HVAI device using a COC syringe. The elastic element used in this study had the same k-elastic constant as the devices used in previous studies using the HVAI (Studies 22148 and 23027). The COC syringe was allowed to reach room temperature for at least 30 minutes prior to final assembly. The syringe cap was removed and replaced with a 25G x 1-inch Becton Dickinson needle, which was then loaded into the fixture where the HVAI was assembled by Halozyme engineers. After final assembly of the HVAI, a needle protector was placed on the needle to achieve an exposed needle depth of 10 mm ± 1 mm. The length of each needle was recorded.

[0211] Animal description

[0212] Species: Pig (domestic pig) Sus scrofa domestica ))

[0213] Variety: Yucatan (small)

[0214] Sex: Female

[0215] Age: Approximately 4-6 months

[0216] Weight: Approximately 18kg-22kg

[0217] Quantity: 3

[0218] Source: Premier BioSource (Ramona, CA)

[0219] Animal husbandry: Animals are kept in steel enclosures equipped with automatic watering systems for free access. They are fed twice daily (morning and afternoon), but fasted after midnight on the day of the study to prevent complications from anesthesia. The room environment is maintained at a temperature of approximately 19°C-23°C and a relative humidity of 40%-70%, using a 12-hour light / 12-hour dark cycle. Animals are allowed at least 3 days to acclimatize in the animal enclosure before the study begins.

[0220] Test materials

[0221] The duration of injection was measured using a handheld stopwatch (Fisher). Any backleakage lasting 30 seconds was collected immediately after administration and weighed using an analytical balance with a sensitivity of 0.1 mg. Qualitative assessment and imaging were acquired immediately after injection.

[0222] Research Procedures

[0223] Before the study began, the animals' general health was assessed and their weight was collected. All anesthesia was administered and monitored. The day before the study, syringes were filled with the appropriate test solution and assembled into HVAIs, then stored at 2°C–8°C. The devices were allowed to acclimatize to room temperature for at least 30 minutes before use and used within 2 hours. After anesthesia, the animals were placed supine on a heated operating table and kept under isoflurane gas for the entire duration of the procedure. Following anesthesia, the abdominal area was cleaned with Nolvasan, followed by wiping the injection sites with gauze containing 70% isopropanol and drying with sterile gauze. Injection sites were located in the left and right abdominal regions, approximately 5 cm cranially from the inguinal fold towards the midline and approximately 3 cm towards the animal's midline. Each injection site was marked with a permanent marker and photographed using a standard camera and a 3D camera before needle insertion. Once the injection was completed, both additional devices were placed at room temperature for use on the next animal.

[0224] Insert the needle vertically into the marked injection site and hold the HVAI in place by hand. The target needle insertion depth is approximately 10 mm. Use a stopwatch to time the injection. Upon completion of the injection, remove the needle and discard the HVAI device. Then, at the injection site, backdrain the test solution into a balanced ophthalmic sponge for 30 seconds and measure the weight of the ophthalmic sponge using an analytical balance. Mark the edge of the injection site wheal with a permanent marker, measure and record the length, width, and height using digital calipers, and then photograph using a standard camera and a 3D camera. Perform caliper measurements and photographs immediately after the injection. Qualitatively assess and grade the severity of erythema, swelling size and appearance, and hardness of the local injection site daily using the scoring systems described in Tables 4, 5, and 6.

[0225]

[0226] Approximately 24 hours after the first injection, each animal was anesthetized again, and the erythema, induration, and swelling at the injection site were assessed. The animals were then photographed and returned to their cages.

[0227] Following an assessment on the third day, the animals were humanely euthanized using injectable euthanasia drugs provided by animal husbandry staff. After euthanasia, full-thickness perforated biopsies (12 mm) were performed at the injection sites and placed in 10% formalin. After obtaining the perforated biopsies, the animal carcasses were removed and disposed of as biohazardous waste. Sections were prepared from the tissue samples, and histopathology was evaluated using hematoxylin and eosin staining. Each injection site (two biopsies) and one untreated site were analyzed. Following horizontal sectioning techniques, three tissue depths were examined from each tissue section, representing the beginning, middle, and end of each section.

[0228] Computational and statistical methods

[0229] Assessment of injection time

[0230] The duration of the injection was measured using a digital stopwatch with a resolution of 0.1 seconds.

[0231] Assessment of localized swelling volume and area using calipers and 3D imaging

[0232] The volume and area of ​​the swelling after injection were measured using both caliper measurements and 3D camera image analysis. For caliper measurements, digital calipers were used to measure the length, width, and height of the wheal formed after injection. Length and width were defined as side-to-side measurements along the longest axis of the wheal (i.e., diameter). These values ​​were recorded manually, and the formula for half an ellipsoid, Vol = (2 / 3), was used. π A B C determines the volume, where A = length / 2, B = width / 2, and C = height.

[0233] 3D imaging was used as a longitudinal method to measure post-injection swelling. By acquiring high-resolution pre- and post-injection 3D images, the distance between two registered surfaces could be determined. A factory-calibrated bifocal imaging system was used to capture images to measure the distance between the surfaces. Surface registration was performed using a multi-point method that utilized common landmarks between the pre- and post-injection images. Proprietary software (Canfield Biosciences, Inc.) was used to calculate the volume, area, and height of post-injection swelling for each injection.

[0234] Caliper measurements and 3D imaging measurements produce different values ​​for volume, area, and wheal height. The difference is a result of variations in wheal size measurements. 3D measurements calculate wheal height based on the distance from the top of the wheal to the original skin location, while caliper measurements measure wheal height from the top of the wheal to its edge. Due to skin curvature, this results in an overall increase in wheal height for caliper measurements compared to 3D measurements, leading to a larger wheal volume and height. However, the measurements are consistent with each other and therefore differ only due to the methodologies.

[0235] Quantitative measurements were taken immediately after injection (T0) and daily thereafter until approximately 72 hours later.

[0236] Duration of injection

[0237] Collect the backleak for 30 seconds after injection using a pre-weighed ophthalmic sponge. Then reweigh the ophthalmic sponge and record the weight of the backleak. Mark the swelling (wheal) at the injection site and measure it using digital calipers. Take pre- and post-injection photographs using a 3D imaging system (Canfield Scientific). This technique allows for point-to-point alignment of the two images via multi-point surface registration. The distance between any two points is then represented using a colorimetric surface profilograph. Areas with no difference between the two images are shown in gray. Where the post-injection image is higher than the pre-injection image, the area is shown in blue shading. Where the post-injection image is lower than the pre-injection image, the distance is shown in orange shading. Color intensity is proportional to the amount of distance measured between the images, with darker blue indicating a larger distance from the pre-injection image. Measurements outside the range (distances greater than 6 mm) are depicted in white. Wheal measurements of volume and height include areas outside the range.

[0238] Each animal has a pre-injection 3D image of the injection site, followed by a second image taken immediately after injection, and these images are mapped to each other using multi-point registration. Using these registered pre-injection / post-injection images, proprietary software (Vectra H1 software; Canfield Sciences) is then used to calculate the volume, height, perimeter, length, and width of each wheal.

[0239] Qualitative assessments were performed immediately after injection (T0) and daily thereafter. No erythema, swelling, or induration was detected on days 1, 2, or 3 post-injection.

[0240]

[0241] Bold data were generated from human clinical trials. When using a 25G-Terumo needle in preclinical trials, the applied force was measured to be 42.6 ± 1.1 (N ± SEM). However, the measured value in clinical trials at the same flow rate was 10% lower. This can be calculated as: % decrease = 100 - (38.7) 100 / 42.6 = -9.15% (approximately 90.85% of the value measured in pig studies) Therefore, the predicted time for HVAI injection using a 25G-Terumo needle will be: 25G-Terumo: 90.85% 19.8 seconds = 18.0 seconds 25G-BDL 90.85% 30.0 seconds = 27.5 seconds Based on these estimates, it is recommended to test the HVAI device using the 25G-BD needle group during injection visit #2 of the Halozyme 104-105 clinical trial. The expected AF value is calculated by multiplying the value from group A of 104-105 by 2, since the Hagen-Poiseuille equation predicts that the pressure will increase proportionally to the flow rate. Therefore, since P=F / A, when the flow rate increases by a factor of 2, the applied force should increase by a factor of 2.

[0242] The expected HVAI injection time was calculated by multiplying the values ​​from preclinical miniature pig studies (in bold) by 90.8%.

[0243] Example 3

[0244] Figures 1A to 1B This data includes information on the injection of 10 mL of GGL+rHuPH20 into miniature pigs using different needle sizes. Needle size selection is used to control injection time. Figure 1A It contains data obtained using a syringe pump. Figure 1B It contains data obtained using HVAI.

[0245] Figure 2 Data from human studies are provided, demonstrating that a miniature pig model can predict the timing of HVAI injections in humans. The data were obtained from a human study (Part B) in which 10 mL was injected over 30 seconds. Comparisons of the forces applied during injection between humans and miniature pigs are provided with scaling factors as shown in the table below.

[0246]

[0247] Data from miniature pig models predicted the timing of HVAI injections in humans. Figure 2 (See the table below). Preclinical data guided the selection of needle gauge for human studies, with a target duration of 30 seconds.

[0248]

[0249] Figures 3A to 3B The data contained demonstrate that a miniature pig model can estimate HVAI injection site responses. Figure 3A Data on injection site responses in miniature pigs were provided. Figure 3B It provides data on human injection site responses.

Claims

1. A method for determining one or more parameters of a formulation injection in a human subject, the method comprising: The preparation containing a therapeutically effective amount of hyaluronidase was administered to animal subjects via subcutaneous injection from a large-volume autoinjector, in a dose of approximately 3 mL to approximately 50 mL. Measure one or more parameters of the subcutaneous injection in the animal subjects; as well as The same one or more parameters for the subcutaneous injection in the human subjects are determined based on one or more measured parameters from the animal subjects; wherein the one or more parameters are selected from injection time, injection force, and injection site response data.

2. The method according to claim 1, wherein the hyaluronidase is a soluble hyaluronidase.

3. The method according to claim 1 or 2, wherein the hyaluronidase is recombinant human hyaluronidase.

4. The method according to any one of claims 1 to 3, wherein the hyaluronidase is recombinant human hyaluronidase PH20.

5. The method according to claim 4, wherein the recombinant human hyaluronidase PH20 enzyme comprises a sequence having at least 95% amino acid sequence identity with the amino acid sequence of SEQ ID NO:

1.

6. The method according to any one of claims 1 to 5, wherein the formulation further comprises a therapeutically effective amount of an active ingredient, said active ingredient being selected from small molecules, proteins, peptide fragments, biological agents, nanoparticles, antibodies, antibody fragments, and small molecule antiviral agents.

7. The method of claim 6, wherein the antibody comprises a monoclonal antibody or a polyclonal antibody.

8. The method according to claim 7, wherein the monoclonal antibody or the polyclonal antibody comprises trastuzumab, rituximab, pertuzumab, atezolizumab, immunoglobulin, daratumumab, egamod α, nivolumab, or ozoglucon.

9. The method of claim 6, wherein the protein comprises γ-globulin.

10. The method according to any one of claims 1 to 9, wherein the animal subject is a miniature pig.

11. The method according to any one of claims 1 to 10, wherein the animal subject is a Yucatan miniature pig.

12. The method according to any one of claims 1 to 11, the method comprising administering about 10 mL to about 20 mL of the preparation to the animal subject.

13. The method according to any one of claims 1 to 9, wherein the method comprises administering about 10 mL to about 20 mL of the formulation to the human subject.

14. The method according to any one of claims 1 to 12, wherein the method comprises administering about 3 mL to about 15 mL of the preparation to the animal subject.

15. The method according to any one of claims 1 to 9 or 13, the method comprising administering about 3 mL to about 15 mL of the formulation to the human subject.

16. The method according to any one of claims 1 to 12 or 14, the method comprising administering to the animal 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, or about 6.5 mL. Approximately 6.6 mL, approximately 6.7 mL, approximately 6.8 mL, approximately 6.9 mL, approximately 7 mL, approximately 7.1 mL, approximately 7.2 mL, approximately 7.3 mL, approximately 7.4 mL, approximately 7.5 mL, approximately 7.6 mL, approximately 7.7 mL, approximately 7.8 mL, approximately 7.9 mL, approximately 8 mL, approximately 8.1 mL, approximately 8.2 mL, approximately 8.3 mL, approximately 8.4 mL, approximately 8.5 mL, approximately 8.6 mL, approximately 8.7 mL, approximately 8.8 mL, approximately 8.9 mL, approximately 9 mL, approximately 9 mL, approximately 9.1 mL, approximately 9.2 mL, approximately 9.3 mL, approximately 9.4 mL, approximately 9.5 mL, approximately 9.6 mL, approximately 9.7 mL, approximately 9.8 mL, approximately 9.9 mL, approximately 10 mL, approximately 10.1 mL, approximately 10 0.2mL, approximately 10.3mL, approximately 10.4mL, approximately 10.5mL, approximately 10.6mL, approximately 10.7mL, approximately 10.8mL, approximately 10.9mL, approximately 11mL, approximately 11.1mL, approximately 11.2mL, approximately 11.3mL, approximately 11.4mL, approximately 11.5mL, approximately 11.6mL, approximately 11.7mL, approximately 11.8mL, approximately 11.9mL, approximately 12mL, approximately 12.1mL, approximately 12.2mL, approximately 12.3mL, approximately 12.4mL, approximately 12.5mL, approximately 12.6mL, approximately 12.7mL, approximately 12.8mL, approximately 12.9mL, approximately 13mL, approximately 13.1mL, approximately 13.2mL, approximately 13.3mL, approximately 13... 0.4mL, approximately 13.5mL, approximately 13.6mL, approximately 13.7mL, approximately 13.8mL, approximately 13.9mL, approximately 14mL, approximately 14.1mL, approximately 14.2mL, approximately 14.3mL, approximately 14.4mL, approximately 14.5mL, approximately 14.6mL, approximately 14.7mL, approximately 14.8mL, approximately 14.9mL, approximately 15mL, approximately 15.1mL, approximately 15.2mL, approximately 15.3mL, approximately 15.4mL, approximately 15.5mL, approximately 15.6mL, approximately 15.7mL, approximately 15.8mL, approximately 15.9mL, approximately 16mL, approximately 16.1mL, approximately 16.2mL, approximately 16.3mL, approximately 16.4mL, approximately 16.5mL, approximately 16.6mL, approximately 16.7mL, approximately 16.8mL, approximately 16.9mL, approximately 17mL, approximately 17.1mL, approximately 17.2mL, approximately 17.3mL, approximately 17.4mL, approximately 17.5mL, approximately 17.6mL, approximately 17.7mL, approximately 17.8mL, approximately 17.9mL, approximately 18mL, approximately 18.1mL, approximately 18.2mL, approximately 18.3mL, approximately 18.4mL, approximately 18.5mL, approximately 18.6mL, approximately 18. 7mL, approximately 18.8mL, approximately 18.9mL, approximately 19mL, approximately 19.1mL, approximately 19.2mL, approximately 19.3mL, approximately 19.4mL, approximately 19.5mL, approximately 19.6mL, approximately 19.7mL, approximately 19.8mL, approximately 19.9mL, approximately 20mL, approximately 20.1mL, approximately 20.2mL, approximately 20.3mL, approximately 20.4mL, approximately 20.5mL, approximately 20.6mL, approximately 20.7mL, approximately 20.8mL mL, approximately 20.9 mL, approximately 21 mL, approximately 21.1 mL, approximately 21.2 mL, approximately 21.3 mL, approximately 21.4 mL, approximately 21.5 mL, approximately 21.6 mL, approximately 21.7 mL, approximately 21.8 mL, approximately 21.9 mL, approximately 22 mL, approximately 22.1 mL, approximately 22.2 mL, approximately 22.3 mL, approximately 22.4 mL, approximately 22.5 mL, approximately 22.6 mL, approximately 22.7 mL, approximately 22.8 mL, approximately 22.9 mL Approximately 23 mL, 23.1 mL, 23.2 mL, 23.3 mL, 23.4 mL, 23.5 mL, 23.6 mL, 23.7 mL, 23.8 mL, 23.9 mL, 24 mL, 24.1 mL, 24.2 mL, 24.3 mL, 24.4 mL, 24.5 mL, 24.6 mL, 24.7 mL, 24.8 mL, 24.9 mL, or 25 mL.

17. The method according to any one of claims 1 to 9, 13, or 15, the method comprising administering to the human 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, or about 6 mL. 0.5mL, approximately 6.6mL, approximately 6.7mL, approximately 6.8mL, approximately 6.9mL, approximately 7mL, approximately 7.1mL, approximately 7.2mL, approximately 7.3mL, approximately 7.4mL, approximately 7.5mL, approximately 7.6mL, approximately 7.7mL, approximately 7.8mL, approximately 7.9mL, approximately 8mL, approximately 8.1mL, approximately 8.2mL, approximately 8.3mL, approximately 8.4mL, approximately 8.5mL, approximately 8.6mL, approximately 8.7mL, approximately 8.8mL, approximately 8.9mL, approximately 9mL, approximately 9mL, approximately 9.1mL, approximately 9.2mL, approximately 9.3mL, approximately 9.4mL, approximately 9.5mL, approximately 9.6mL, approximately 9.7mL, approximately 9.8mL, approximately 9.9mL, approximately 10mL, approximately 10.1mL L, approximately 10.2 mL, approximately 10.3 mL, approximately 10.4 mL, approximately 10.5 mL, approximately 10.6 mL, approximately 10.7 mL, approximately 10.8 mL, approximately 10.9 mL, approximately 11 mL, approximately 11.1 mL, approximately 11.2 mL, approximately 11.3 mL, approximately 11.4 mL, approximately 11.5 mL, approximately 11.6 mL, approximately 11.7 mL, approximately 11.8 mL, approximately 11.9 mL, approximately 12 mL, approximately 12.1 mL, approximately 12.2 mL, approximately 12.3 mL, approximately 12.4 mL, approximately 12.5 mL, approximately 12.6 mL, approximately 12.7 mL, approximately 12.8 mL, approximately 12.9 mL, approximately 13 mL, approximately 13.1 mL, approximately 13.2 mL, approximately 13.3 mL mL, approximately 13.4 mL, approximately 13.5 mL, approximately 13.6 mL, approximately 13.7 mL, approximately 13.8 mL, approximately 13.9 mL, approximately 14 mL, approximately 14.1 mL, approximately 14.2 mL, approximately 14.3 mL, approximately 14.4 mL, approximately 14.5 mL, approximately 14.6 mL, approximately 14.7 mL, approximately 14.8 mL, approximately 14.9 mL, approximately 15 mL, approximately 15.1 mL, approximately 15.2 mL, approximately 15.3 mL, approximately 15.4 mL, approximately 15.5 mL, approximately 15.6 mL, approximately 15.7 mL, approximately 15.8 mL, approximately 15.9 mL, approximately 16 mL, approximately 16.1 mL, approximately 16.2 mL, approximately 16.3 mL, approximately 16.4 mL, approximately 16.5mL, approximately 16.6mL, approximately 16.7mL, approximately 16.8mL, approximately 16.9mL, approximately 17mL, approximately 17.1mL, approximately 17.2mL, approximately 17.3mL, approximately 17.4mL, approximately 17.5mL, approximately 17.6mL, approximately 17.7mL, approximately 17.8mL, approximately 17.9mL, approximately 18mL, approximately 18.1mL, approximately 18.2mL, approximately 18.3mL, approximately 18.4mL, approximately 18.5mL, approximately 18.6mL L, approximately 18.7 mL, approximately 18.8 mL, approximately 18.9 mL, approximately 19 mL, approximately 19.1 mL, approximately 19.2 mL, approximately 19.3 mL, approximately 19.4 mL, approximately 19.5 mL, approximately 19.6 mL, approximately 19.7 mL, approximately 19.8 mL, approximately 19.9 mL, approximately 20 mL, approximately 20.1 mL, approximately 20.2 mL, approximately 20.3 mL, approximately 20.4 mL, approximately 20.5 mL, approximately 20.6 mL, approximately 20.7 mL, approximately 20.8 mL, approximately 20.9 mL, approximately 21 mL, approximately 21.1 mL, approximately 21.2 mL, approximately 21.3 mL, approximately 21.4 mL, approximately 21.5 mL, approximately 21.6 mL, approximately 21.7 mL, approximately 21.8 mL, approximately 21.9 mL, approximately 22 mL, approximately 22.1 mL, approximately 22.2 mL, approximately 22.3 mL, approximately 22.4 mL, approximately 22.5 mL, approximately 22.6 mL, approximately 22.7 mL, approximately 22.8 mL, approximately 22. 9 mL, approximately 23 mL, approximately 23.1 mL, approximately 23.2 mL, approximately 23.3 mL, approximately 23.4 mL, approximately 23.5 mL, approximately 23.6 mL, approximately 23.7 mL, approximately 23.8 mL, approximately 23.9 mL, approximately 24 mL, approximately 24.1 mL, approximately 24.2 mL, approximately 24.3 mL, approximately 24.4 mL, approximately 24.5 mL, approximately 24.6 mL, approximately 24.7 mL, approximately 24.8 mL, approximately 24.9 mL, or approximately 25 mL.

18. The method according to any one of claims 1 to 17, wherein the formulation is in a pre-filled syringe.

19. The method of claim 18, wherein the pre-filled 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, about 6.4 mL, about 6.5 mL, about 6.6 mL, about 6.7 mL. mL, approximately 6.8 mL, approximately 6.9 mL, approximately 7 mL, approximately 7.1 mL, approximately 7.2 mL, approximately 7.3 mL, approximately 7.4 mL, approximately 7.5 mL, approximately 7.6 mL, approximately 7.7 mL, approximately 7.8 mL, approximately 7.9 mL, approximately 8 mL, approximately 8.1 mL, approximately 8.2 mL, approximately 8.3 mL, approximately 8.4 mL, approximately 8.5 mL, approximately 8.6 mL, approximately 8.7 mL, approximately 8.8 mL, approximately 8.9 mL, approximately 9 mL, approximately 9.1 mL, approximately 9.2 mL, approximately 9.3 mL, approximately 9.4 mL, approximately 9.5 mL, approximately 9.6 mL, approximately 9.7 mL, approximately 9.8 mL, approximately 9.9 mL, approximately 10 mL, approximately 10.1 mL, approximately 10.2 mL, approximately 10.3 mL L, approximately 10.4 mL, approximately 10.5 mL, approximately 10.6 mL, approximately 10.7 mL, approximately 10.8 mL, approximately 10.9 mL, approximately 11 mL, approximately 11.1 mL, approximately 11.2 mL, approximately 11.3 mL, approximately 11.4 mL, approximately 11.5 mL, approximately 11.6 mL, approximately 11.7 mL, approximately 11.8 mL, approximately 11.9 mL, approximately 12 mL, approximately 12.1 mL, approximately 12.2 mL, approximately 12.3 mL, approximately 12.4 mL, approximately 12.5 mL, approximately 12.6 mL, approximately 12.7 mL, approximately 12.8 mL, approximately 12.9 mL, approximately 13 mL, approximately 13.1 mL, approximately 13.2 mL, approximately 13.3 mL, approximately 13.4 mL, approximately 13.5 mL mL, approximately 13.6 mL, approximately 13.7 mL, approximately 13.8 mL, approximately 13.9 mL, approximately 14 mL, approximately 14.1 mL, approximately 14.2 mL, approximately 14.3 mL, approximately 14.4 mL, approximately 14.5 mL, approximately 14.6 mL, approximately 14.7 mL, approximately 14.8 mL, approximately 14.9 mL, approximately 15 mL, approximately 15.1 mL, approximately 15.2 mL, approximately 15.3 mL, approximately 15.4 mL, approximately 15.5 mL, approximately 15.6 mL, approximately 15.7 mL, approximately 15.8 mL, approximately 15.9 mL, approximately 16 mL, approximately 16.1 mL, approximately 16.2 mL, approximately 16.3 mL, approximately 16.4 mL, approximately 16.5 mL, approximately 16.6 mL, approximately 16.7mL, approximately 16.8mL, approximately 16.9mL, approximately 17mL, approximately 17.1mL, approximately 17.2mL, approximately 17.3mL, approximately 17.4mL, approximately 17.5mL, approximately 17.6mL, approximately 17.7mL, approximately 17.8mL, approximately 17.9mL, approximately 18mL, approximately 18.1mL, approximately 18.2mL, approximately 18.3mL, approximately 18.4mL, approximately 18.5mL, approximately 18.6mL, approximately 18.7mL, approximately 18. 8mL, approximately 18.9mL, approximately 19mL, approximately 19.1mL, approximately 19.2mL, approximately 19.3mL, approximately 19.4mL, approximately 19.5mL, approximately 19.6mL, approximately 19.7mL, approximately 19.8mL, approximately 19.9mL, approximately 20mL, approximately 20.1mL, approximately 20.2mL, approximately 20.3mL, approximately 20.4mL, approximately 20.5mL, approximately 20.6mL, approximately 20.7mL, approximately 20.8mL, approximately 20. 9mL, approximately 21mL, approximately 21.1mL, approximately 21.2mL, approximately 21.3mL, approximately 21.4mL, approximately 21.5mL, approximately 21.6mL, approximately 21.7mL, approximately 21.8mL, approximately 21.9mL, approximately 22mL, approximately 22.1mL, approximately 22.2mL, approximately 22.3mL, approximately 22.4mL, approximately 22.5mL, approximately 22.6mL, approximately 22.7mL, approximately 22.8mL, approximately 22.9mL, approximately 23mL The preparation may be expressed in L, approximately 23.1 mL, approximately 23.2 mL, approximately 23.3 mL, approximately 23.4 mL, approximately 23.5 mL, approximately 23.6 mL, approximately 23.7 mL, approximately 23.8 mL, approximately 23.9 mL, approximately 24 mL, approximately 24.1 mL, approximately 24.2 mL, approximately 24.3 mL, approximately 24.4 mL, approximately 24.5 mL, approximately 24.6 mL, approximately 24.7 mL, approximately 24.8 mL, approximately 24.9 mL, or approximately 25 mL.

20. The method of claim 18 or 19, wherein the pre-filled syringe comprises a needle of about 20 to about 33 gauge.

21. The method according to any one of claims 18 to 20, wherein the pre-filled 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.

22. The method according to any one of claims 1 to 12, 14 or 16, the method comprising administering the formulation to the animal subject at a rate of about 0.08 mL / s to about 1.00 mL / s.

23. The method according to any one of claims 1 to 9, 13, 15 or 17, the method comprising administering the formulation to the human subject at a rate of about 0.08 mL / s to about 1.00 mL / s.

24. The method according to any one of claims 1 to 12, 14, 16 or 22, the method comprising administering the formulation to the animal subject at a rate of at least about 0.08 mL / s to about 1.0 mL / s.

25. The method according to any one of claims 1 to 9, 13, 15, 17 or 23, the method comprising administering the formulation to the human subject at a rate of at least about 0.08 mL / s to about 1.0 mL / s.

26. The method according to any one of claims 1 to 12, 14 or 16, the method comprising administering the formulation to the animal subject at a rate of at least or faster than about 0.08 mL / s to about 1.00 mL / s.

27. The method according to any one of claims 1 to 9, 13, 15 or 17, the method comprising administering the formulation to the human subject at a rate of at least or faster than about 0.08 mL / s to about 1.00 mL / s.

28. The method according to any one of claims 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes about 10 seconds to about 40 seconds.

29. The method according to any one of claims 1 to 9, 13, 15, 17, 23 or 25, wherein administering the formulation to the human subject takes about 10 seconds to about 40 seconds.

30. The method according to any one of claims 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes at least about 10 seconds to about 40 seconds.

31. The method according to any one of claims 1 to 9, 13, 15, 17, 23 or 25, wherein administering the formulation to the human subject takes at least about 10 seconds to about 40 seconds.

32. The method according to any one of claims 1 to 12, 14, 16, 22 or 24, wherein administering the formulation to the animal subject takes at least or less than about 10 seconds to about 40 seconds.

33. The method according to any one of claims 1 to 9, 13, 15, 17, 23 or 25, wherein administering the formulation to the human subject takes at least or less than about 10 seconds to about 40 seconds.

34. The method according to any one of claims 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes about 15 seconds to about 30 seconds.

35. The method according to any one of claims 1 to 9, 13, 15, 17, 23 or 25, wherein administering the formulation to the human subject takes about 15 seconds to about 30 seconds.

36. The method according to any one of claims 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes at least about 15 seconds to about 30 seconds.

37. The method according to any one of claims 1 to 9, 13, 15, 17, 23 or 25, wherein administering the formulation to the human subject takes at least about 15 seconds to about 30 seconds.

38. The method according to any one of claims 1 to 12, 14, 16, 22 or 24, wherein the administration of the formulation to the animal subject takes at least or less than about 15 seconds to about 30 seconds.

39. The method according to any one of claims 1 to 9, 13, 15, 17, 23 or 25, wherein administering the formulation to the human subject takes at least or less than about 15 seconds to about 30 seconds.

40. The method according to any one of claims 1 to 12, 14, 16, 22 or 24, the method comprising administering about 10 mL of the formulation to the animal subject at a rate of about 0.32 mL / s to about 0.42 mL / s.

41. The method according to any one of claims 1 to 9, 13, 15, 17, 23 or 25, the method comprising administering about 10 mL of the formulation to the human subject at a rate of about 0.32 mL / s to about 0.42 mL / s.

42. The method according to any one of claims 1 to 41, wherein the parameter is injection force.

43. The method of claim 42, wherein the animal subject is a miniature pig.

44. The method of claim 42 or 43, wherein the animal subject is a Yucatan miniature pig.

45. The method according to any one of claims 42 to 44, wherein the formulation further comprises γ-globulin.

46. ​​The method according to any one of claims 42 to 45, wherein the subcutaneous administration to the animal subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

47. The method of claim 42 or 45, wherein the subcutaneous administration to the human subject is performed via a high-volume autoinjector equipped with a 25-gauge needle, optionally via a high-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

48. The method of any one of claims 42 to 47, the method comprising determining the injection force of the subcutaneous injection in the human subject by multiplying the measured injection force of the animal subject by a scaling factor.

49. The method of claim 48, wherein the scaling factor is about 0.80 to about 0.

99.

50. The method of claim 48 or 49, wherein the scaling factor is about 0.85 to about 0.95; optionally about 0.88 to about 0.

92.

51. The method according to any one of claims 42 to 50, wherein the injection force is measured in Newtons (N).

52. The method according to any one of claims 42 to 51, the method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.42 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. The injection force of the subcutaneous injection in the miniature pig was measured to be approximately 25 N to approximately 50 N; as well as The injection force for the same subcutaneous injection in the human subjects was determined by multiplying the injection force in the miniature pigs by a scaling factor of about 0.85 to about 0.95, thereby obtaining an estimated injection force of about 21 N to about 48 N.

53. The method according to any one of claims 42 to 52, the method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. The injection force of the subcutaneous injection in the miniature pig was measured to be approximately 40 N to approximately 46 N; and An estimated injection force of about 35 N to about 42 N was obtained by multiplying the injection force in the miniature pigs by a scaling factor of about 0.88 to about 0.92 to determine the same subcutaneous injection force in the human subjects.

54. The method according to any one of claims 1 to 41, wherein the parameter is the injection time.

55. The method of claim 54, wherein the animal subject is a miniature pig.

56. The method of claim 54 or 55, wherein the animal subject is a Yucatan miniature pig.

57. The method according to any one of claims 54 to 56, wherein the formulation further comprises γ-globulin.

58. The method according to any one of claims 54 to 57, wherein the subcutaneous administration to the animal subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

59. The method of claim 54 or 57, wherein the subcutaneous administration to the human subject is performed via a high-volume autoinjector equipped with a 25-gauge needle, optionally via a high-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

60. The method of any one of claims 54 to 59, the method comprising determining the injection time of the subcutaneous injection in the human subject by multiplying the measured injection time of the animal subject by a scaling factor.

61. The method of claim 60, wherein the scaling factor is about 0.80 to about 0.

99.

62. The method of claim 60 or 61, wherein the scaling factor is from about 0.85 to about 0.95; optionally from about 0.88 to about 0.

92.

63. The method according to any one of claims 54 to 62, wherein the injection time is measured in seconds.

64. The method according to any one of claims 54 to 63, the method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. The injection time for the subcutaneous injection in the miniature pigs was measured to be approximately 15 seconds to approximately 35 seconds; and The injection time for the same subcutaneous injection in the human subjects was determined by multiplying the injection time in the miniature pigs by a scaling factor of about 0.85 to about 0.95, thereby obtaining an estimated injection time of about 13 seconds to about 33 seconds.

65. The method according to any one of claims 54 to 64, the method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. The injection time for the subcutaneous injection in the miniature pigs was measured to be approximately 18 seconds to approximately 30 seconds; and The injection time for the same subcutaneous injection in the human subjects was determined by multiplying the injection time in the miniature pigs by a scaling factor of about 0.88 to about 0.92, thereby obtaining an estimated injection time of about 16 seconds to about 28 seconds.

66. The method according to any one of claims 1 to 41, wherein the parameter is injection site reaction data.

67. The method of claim 66, wherein the injection site reaction data includes one or more of the following: backflow at the injection site, swelling volume at the injection site, swelling height at the injection site, swelling size of the wheal at the injection site, induration of the wheal at the injection site, and wheal regression at the injection site.

68. The method of claim 66 or 67, wherein the animal subject is a miniature pig.

69. The method according to any one of claims 66 to 68, wherein the animal subject is a Yucatan miniature pig.

70. The method according to any one of claims 66 to 69, wherein the formulation further comprises γ-globulin.

71. The method according to any one of claims 66 to 70, wherein the subcutaneous administration to the animal subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

72. The method according to any one of claims 66, 67 or 70, wherein the subcutaneous administration to the human subject is performed via a large-volume autoinjector equipped with a 25-gauge needle, optionally via a large-volume autoinjector dispensing the formulation from a pre-filled syringe equipped with a 25-gauge needle.

73. The method according to any one of claims 66 to 72, the method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.42 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. Measure the response at one or more injection sites of the subcutaneous injection in the miniature pig and assign a score of 0, 1, 2, 3 or 4; as well as It is determined that responses at one or more injection sites for the same subcutaneous injection in the human subjects will have the same score.

74. The method according to any one of claims 66 to 73, the method comprising: Approximately 10 mL of the formulation was administered to a small pig animal subject via subcutaneous injection at a rate of approximately 0.30 mL / s to approximately 0.36 mL / s, wherein the formulation was dispensed from a pre-filled syringe equipped with a 25-gauge needle using a high-volume autoinjector. Measure the response at one or more injection sites of the subcutaneous injection in the miniature pig and assign a score of 0, 1, 2, 3 or 4; as well as It is determined that responses at one or more injection sites for the same subcutaneous injection in the human subjects will have the same score.

75. The method according to any one of claims 66 to 71, 73 or 74, wherein the subcutaneous administration to the animal subject comprises a single injection.

76. The method according to any one of claims 66, 67, 70 or 72 to 74, wherein the subcutaneous administration to the human subject comprises a single injection.

77. The method according to any one of claims 66 to 71, 73 or 75, wherein the subcutaneous administration to the animal subject comprises two or more injections.

78. The method according to any one of claims 66, 67, 69 or 71 to 73, wherein the subcutaneous administration to the human subject comprises two or more injections.

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