Autoinjectors and related methods of use

Abstract

An injection device may include: a housing; the container is arranged in the shell, and the container is used for packaging fluid and is provided with a first end and a second end; a conduit movable relative to the container, where the conduit is not in fluid communication with the fluid encapsulated by the container when in a first position and is in fluid communication with the fluid encapsulated by the container when in a second position and configured to deliver the fluid from the container to a patient; and a lock, the lock being removable from the housing, the lock having a first portion and a second portion.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 502,883, filed May 17, 2023, which is incorporated herein by reference in its entirety. Technical Field

[0003] This disclosure relates to an autoinjector and related methods of use.

[0004] introduction

[0005] In most available autoinjectors, the needle is deployed when activated by the user, and fluid is delivered from the needle into the user's body. After fluid delivery is complete, the needle can be retracted for user comfort, needle safety, and a positive impression of the product. However, many autoinjectors can be accidentally triggered by drops or vibrations. Additionally, many autoinjectors may lack suitable control logic for stopping injection when appropriate. Summary of the Invention

[0006] In one aspect, this disclosure relates to an injection device. The injection device may include: a housing; a container disposed within the housing, the container encapsulating a fluid and having a first end and a second end; a catheter movable relative to the container, wherein the catheter is not in fluid communication with the fluid encapsulated by the container in a first position, and is in fluid communication with the fluid encapsulated by the container in a second position and is configured to deliver fluid from the container to a patient; and a lock removable from the housing, the lock having a first portion and a second portion. In a first configuration where the lock is coupled to the housing, the first portion of the lock may be disposed outside the housing, and the second portion of the lock may be disposed inside the housing, between the container and the catheter; in the first configuration, the second portion of the lock may prevent the catheter from moving into fluid communication with the fluid encapsulated by the container; and in a second configuration where the lock is removed from the injection device, the catheter may be movable into fluid communication with the fluid encapsulated by the container.

[0007] On the other hand, the injection device may include: a housing; a plunger coupled to and movable relative to the housing; and one or more electronic components used during an injection performed by the injection device, the one or more electronic components being formed within a circuit. In a first configuration, a first portion of the plunger may be disposed within the housing, the circuit may be disconnected, and the one or more electronic components may be in a low-power sleep mode; in a second configuration, the plunger may be movable outward relative to the housing, and the first portion of the plunger may extend outside the housing; and in the second configuration, the circuit may be closed, and the one or more electronic components may transition from the low-power sleep mode to an active mode.

[0008] On the other hand, the injection device may include: a housing, wherein the housing includes a curved bottom surface that is concave when viewed from a point outside the housing, the point being closer to the bottom surface of the housing than the top surface of the housing; a circuit board positioned adjacent to the bottom surface of the housing, wherein the circuit board includes a skin sensor configured to sense the presence of skin in contact with the bottom surface of the housing; and a controller coupled to the circuit board, wherein the controller is configured to initiate injection by the injection device only after the skin sensor senses the presence of skin in contact with the bottom surface of the housing.

[0009] In another aspect, this disclosure relates to a method of manufacturing an injection device. The method may include: depositing a first material onto a mold, the first material having a first opacity; depositing a second material around the mold and the first material, the second material having a second opacity higher than the first opacity; and positioning a container for encapsulating a drug substance within the injection device and adjacent to a first portion of the injection device formed of the first material.

[0010] On the other hand, the injection device may include: a container disposed within a housing, the container having a first end and a second end; a piston configured to move from the first end of the container toward the second end of the container to dispense a drug from the container; a drive member configured to drive the piston through the container; a transmitter configured to emit a light beam toward the container; a detector positioned on the side of the container opposite the transmitter, wherein the detector is configured to receive the light beam emitted from the transmitter; and a controller coupled to the drive member, the transmitter, and the detector. The controller may be configured to: receive a first signal from the detector when the transmitter is off, the first signal corresponding to the ambient light level around the injection device; receive a second signal from the detector when the transmitter is on; calculate a difference between the light values ​​represented by the first signal and the second signal; and stop operation of the drive member when the difference is less than a threshold.

[0011] On the other hand, the injection device may include: a container disposed within a housing, the container having a first end and a second end; a piston configured to move from the first end of the container toward the second end of the container to dispense a drug from the container; a drive member configured to drive the piston through the container; an emitter configured to emit a light beam toward the container; a detector positioned on the side of the container opposite to the emitter, wherein the detector is configured to receive the light beam emitted from the emitter; and a controller coupled to the drive member, the emitter, and the detector. The controller may be configured to: activate the drive member and the emitter; receive a first signal from the detector when the emitter is activated, the first signal indicating the amount of light received by the detector; allow operation of the drive member to continue for a first time period immediately after activation; and stop operation of the drive member when it is determined that (1) the amount of light received by the detector is less than a first threshold light value and (2) the current of the drive member is greater than a first threshold current value before the amount of light received by the detector subsequently rises to or above the first threshold light value.

[0012] On the other hand, the injection device may include: a container disposed within a housing, having a first end and a second end; a piston configured to move from the first end of the container toward the second end of the container to dispense a drug from the container; a drive member configured to drive the piston through the container; and a controller coupled to the drive member. The controller may be configured to: maintain the speed of the drive member until the current of the drive member exceeds a first threshold; and after the current of the drive member exceeds the first threshold, reduce the voltage of the drive member to maintain the current of the drive member below a second threshold that is greater than or equal to the first threshold. Attached Figure Description

[0013] Various examples are illustrated in conjunction with the accompanying drawings, which are incorporated in and form part of this specification, and together with the specification are used to explain the principles of the disclosed examples and embodiments.

[0014] Various aspects of this disclosure can be implemented in conjunction with the embodiments shown in the accompanying drawings. These drawings illustrate different aspects of this disclosure, and where appropriate, reference numerals showing similar structures, components, materials, and / or elements in different drawings are similarly labeled. It should be understood that various combinations of structures, components, and / or elements are contemplated and are within the scope of this disclosure, in addition to the structures, components, and / or elements specifically shown.

[0015] Furthermore, numerous embodiments are described and illustrated herein. This disclosure is neither limited to any single aspect or embodiment thereof, nor to any combination and / or arrangement of such aspects and / or embodiments. Moreover, each aspect of this disclosure and / or its embodiments may be used alone or in combination with other aspects of this disclosure and / or one or more of its embodiments. For the sake of brevity, certain arrangements and combinations are not discussed and / or illustrated separately herein. It is important to note that embodiments or specific implementations described herein as “exemplary” should not be construed as being more preferred or advantageous than other embodiments or specific implementations, for example; rather, they are intended to reflect or indicate that such embodiment is an “example” embodiment.

[0016] Figure 1 This is a perspective view of an autoinjector according to an example of this disclosure.

[0017] Figure 1A This is a perspective view of a portion of the housing of an autoinjector according to this disclosure.

[0018] Figure 1B This is a perspective view of a portion of the housing of an autoinjector according to this disclosure.

[0019] Figure 2 This is a bottom view of the autoinjector according to this disclosure.

[0020] Figure 3 The side view of the autoinjector according to this disclosure shows the activation switch extending away from the surface facing the tissue.

[0021] Figure 3A The cross-sectional view of the autoinjector according to this disclosure shows the activation switch extending away from the surface facing the tissue.

[0022] Figure 3B The cross-sectional view of the autoinjector according to this disclosure shows the activation switch in a partially pressed position.

[0023] Figure 3C The diagram shows a cross-sectional view of an autoinjector according to this disclosure, illustrating the activation switch in the fully pressed position.

[0024] Figure 4 This is an exploded view of the autoinjector according to this disclosure.

[0025] Figure 4A This is a schematic diagram of the control system of the autoinjector according to the present disclosure.

[0026] Figure 4B This is an exploded view of the autoinjector according to this disclosure.

[0027] Figure 4CThis is a perspective view of a portion of the housing and electronic board according to one aspect of this disclosure.

[0028] Figure 5 It is an exploded view of the needle mechanism according to this disclosure.

[0029] Figure 5A This is a perspective view of the fluid conduit according to this disclosure.

[0030] Figure 5B This is a cross-sectional view of the needle of the fluid conduit according to this disclosure.

[0031] Figure 6 Based on this disclosure Figure 5 A perspective view of the needle mechanism in its first position.

[0032] Figures 7 to 11 yes Figure 5 Side view of the needle mechanism.

[0033] Figure 12 This is a side sectional view of an autoinjector according to this disclosure.

[0034] Figure 13 This is a side sectional view of the puncture mechanism according to this disclosure.

[0035] Figure 13B This is a side sectional view of the autoinjector according to the present disclosure.

[0036] Figure 14 This is a side sectional view of the puncture mechanism according to this disclosure.

[0037] Figure 14A This is a side view of the outer screw of the screw assembly according to this disclosure.

[0038] Figure 14B This is a bottom perspective view of the outer screw of the screw assembly according to this disclosure.

[0039] Figure 14C This is a top perspective view of the outer screw of the screw assembly according to this disclosure.

[0040] Figure 14D Based on this disclosure Figure 14A A cross-sectional view along line AA of the external screw of the screw assembly shown.

[0041] Figure 15 This is a side view of the pin insertion switch according to this disclosure.

[0042] Figure 16A This is a perspective view of a lock for an autoinjector according to one aspect of this disclosure.

[0043] Figure 16B This is a bottom view of the autoinjector and lock according to this disclosure.

[0044] Figure 16C This is a cross-sectional view of the autoinjector and lock according to this disclosure.

[0045] Figure 16D This is a cross-sectional view of the autoinjector and lock according to this disclosure.

[0046] Figure 16E This is a perspective view of a lock for an autoinjector according to one aspect of this disclosure.

[0047] Figure 16F This is a side view of a lock for an autoinjector according to one aspect of this disclosure.

[0048] Figure 17 This is a bottom view of an electronic board for an autoinjector according to the present disclosure.

[0049] Figure 17A This is a perspective view of an electronic board for an autoinjector according to the present disclosure.

[0050] Figures 18 to 20 A flowchart illustrating an exemplary method according to this disclosure is provided.

[0051] Figure 20A and Figure 20B A graph relating to the electrical control of an autoinjector according to this disclosure is depicted.

[0052] Figures 21 to 23 A flowchart illustrating an exemplary method according to this disclosure is provided.

[0053] Figure 24 This is a schematic diagram of the luciferase-based PD-1 bioassay described in Example 8 of this document. Figure A: Unactivated Jurkat cells; Figure B: Jurkat cells activated by aggregation of CD3×CD20 bispecific antibody via T cell receptor (TCR); Figure C: PD-1 activation attenuates the response in activated Jurkat cells; Figure D: Blocking PD-1 restores the response in activated Jurkat cells.

[0054] Figure 25The results show tumor growth and survival in mice implanted with colon-26 tumor cells on day 0 and treated with injections of the molecular combination shown on days 3, 6, 10, 13, and 19 (“Early Treatment Tumor Model”). The figure depicts tumor volume (in mm3) at different time points post-implantation in different experimental groups. Upward arrows along the X-axis indicate the timing of treatment injections. “mlgG2a” is the IgG2 isotype control; “Fc” is the human Fc control; “VEGF Trap” is aflibercept; “anti-PD-1” is the anti-mouse PD-1 clone RPMI-14; and “anti-PD-L1” is the anti-PD-L1 monoclonal antibody as described elsewhere in this document.

[0055] Figure 26 The results of tumor growth and survival in mice implanted with colon-26 tumor cells on day 0 and treated by injection of the molecular combination shown on days 3, 6, 10, 13, and 19 (“Early Treatment Tumor Model”) are shown. The figure shows the tumor volume (in mm3) of each mouse in each experimental group on day 28 post-implantation. “mlgG2a” is an IgG2 isotype control; “Fc” is a human Fc control; “VEGF Trap” is aflibercept; “anti-PD-1” is an anti-mouse PD-1 clone RPMI-14; and “anti-PD-L1” is an anti-PD-L1 monoclonal antibody as described elsewhere in this document.

[0056] Figure 27 This table shows the effect of pH on the stability of 150 mg / mL mAb1 after incubation at 45°C for 28 days. The SE-UPLC and CEX-UPLC “Starting Materials” results are averages of the starting materials for all formulations.

[0057] Figure 28A , Figure 28B and Figure 28C The storage stability of three formulations, F1, F2, and F3, is shown. F1 contains 210 mg / mL mAb1, 10 mM histidine, and 3% proline, at pH 6.0; F2 contains 210 mg / mL mAb1, 10 mM histidine, and 3% sucrose, at pH 6.0; and F3 contains 210 mg / mL mAb1, 10 mM histidine, and 5% sucrose, at pH 6.0. Storage stability was assessed at -80°C (…). Figure 28A -30℃ Figure 28B ) and -20℃ Figure 28C The percentage of high molecular weight substances (HMW) generated after storage for up to 9 months was measured and analyzed by size exclusion chromatography (SEC).

[0058] Figure 29A , Figure 29B and Figure 29CThe storage stability of three formulations, F1, F2, and F3, is shown. F1 contains 150 mg / mL mAb1, 10 mM histidine, 9% sucrose, and 0.2% polysorbate 80 (PS80), pH 6.0; F2 contains 175 mg / mL mAb1, 10 mM histidine, 3% proline, and 0.2% PS80, pH 6.0; and F3 contains 175 mg / mL mAb1, 10 mM histidine, 5% sucrose, 1.5% proline, and 0.2% PS80, pH 6.0. Storage stability was assessed at -80°C (…). Figure 29A -30℃ Figure 29B ) and -20℃ Figure 29C The percentage of high molecular weight substances (HMW) produced after storage for up to 6 months was measured and analyzed by size exclusion chromatography (SEC).

[0059] Figure 30 This is a table showing the viscosity of 150 mg / mL mAb1 with added excipients and viscosity modifiers.

[0060] Figure 31 This table shows the effect of viscosity modifiers on the stability of 175 mg / mL mAb1 after incubation at 45 °C for 14 days. The pH, SE-UPLC, and CEX-UPLC “Starting Material” results are averages of the starting materials for all formulations.

[0061] Similarly, numerous embodiments are described and illustrated herein. This disclosure is neither limited to any single aspect or embodiment thereof, nor to any combination and / or arrangement of such aspects and / or embodiments. Each aspect of this disclosure and / or its embodiments may be used alone or in combination with other aspects of this disclosure and / or its embodiments. For the sake of brevity, the many combinations and arrangements thereof are not discussed separately herein.

[0062] It is worth noting that, for simplicity and clarity, certain aspects of the accompanying drawings depict the general structure and / or construction of various embodiments. Descriptions and details of well-known features and technologies may be omitted to avoid unnecessarily obscuring other features. Elements in the drawings are not necessarily drawn to scale; the dimensions of some features may be exaggerated relative to other elements to better understand the exemplary embodiments. For example, those skilled in the art will understand that sectional views are not drawn to scale and should not be considered as representing proportional relationships between different components. Sectional views are provided to aid in illustrating the various components of the depicted assembly and to show their relative positioning to each other. Detailed Implementation

[0063] Reference will now be made in detail to examples of this disclosure shown in the accompanying drawings. Wherever possible, the same reference numerals will be used throughout the drawings to refer to the same or similar parts. In the following discussion, relative terms such as “about,” “substantially,” and “approximately” are used to indicate possible variations of ±10% in the stated values.

[0064] As mentioned above, existing autoinjectors may be accidentally triggered upon dropping or vibration. Furthermore, existing autoinjectors may lack suitable control logic for stopping injection at the appropriate time. These drawbacks can lead to premature drug deployment, increased complexity of self-administration, introduction of user error, and user discomfort. Therefore, this disclosure relates to various embodiments of injection devices (e.g., autoinjectors) for user-administered medications or other therapeutic agents. Specifically, according to certain embodiments, the likelihood of accidental triggering of the autoinjector can be reduced, and the autoinjector can be further incorporating control logic that improves the operation and user experience of the autoinjector.

[0065] Further details of the autoinjector according to this disclosure can be found in PCT / US2018 / 031077, filed November 8, 2018, by Arnott et al., and published as WO 2018 / 204779 A1, and U.S. Application No. 18 / 055,895, filed November 16, 2022, by Grygus, the entire contents of which are incorporated herein by reference. Further details of the vial puncture system according to this disclosure can be found in U.S. Patent No. 10,182,969, filed March 10, 2016, the entire contents of which are incorporated herein by reference.

[0066] Overall system

[0067] An example of this auto-injector 2 is in Figure 1 , Figure 2 and Figure 3 As shown in the image. Figure 1 As shown, the autoinjector 2 may include a housing 3 having a tissue-engaging (e.g., bottom) surface 4 through which the needle can be deployed and retracted. Figure 1 and Figure 1A As shown, the housing 3 may include a transparent window 50. The transparent window 50 allows an observer to see one or more displays or the interior of the autoinjector 2 and its components, such as the main container and / or the drug product stored in the main container.

[0068] In some implementation schemes, and as such Figure 1As shown, the autoinjector 2 may include a plurality of openings 51 configured to facilitate the propagation of sound generated within the housing 3 (e.g., by a speaker). The autoinjector 2 may have any suitable size to allow for user portability and self-installation. In one example, the autoinjector 2 may have a length of approximately 2.98 inches, a width of approximately 2.07 inches, and a height of approximately 1.07 inches. However, other suitable values ​​may also be used, including, for example, a length of approximately 0.5 inches to approximately 5.0 inches, a width of approximately 0.5 inches to approximately 3.0 inches, and a height of approximately 0.5 inches to approximately 2.0 inches.

[0069] The autoinjector 2 can be oriented about a longitudinal axis 40 (e.g., the X-axis), a transverse axis 42 (e.g., the Y-axis) that is substantially perpendicular to the longitudinal axis 40, and a vertical axis 44 (e.g., the Z-axis) that is substantially perpendicular to both the longitudinal axis 40 and the transverse axis 42.

[0070] like Figure 1 As shown, the adhesive patch 12 can be attached to the tissue-contacting surface 4 to help secure the autoinjector 2 to the user's body (e.g., skin). The adhesive patch 12 can be formed of fabric or any other suitable material and can include an adhesive. For example, the adhesive can be a water-based or solvent-based adhesive, or it can be a hot-melt adhesive. Suitable adhesives also include acrylic-based adhesives, dextrin-based adhesives, and urethane-based adhesives, as well as natural and synthetic elastomers. In some instances, the adhesive provided on the patch 12 can be activated upon contact with the user's skin. In yet another instance, the patch 12 can comprise a nonwoven polyester substrate and an acrylic or silicone adhesive. The patch 12 can be attached to the housing 3 by, for example, double-sided tape or by other mechanisms such as ultrasonic welding. The patch 12 can have a length dimension greater than the width of the autoinjector 2.

[0071] like Figure 2 As shown, the autoinjector 2 may include an opening 6 through which the needle can be deployed and retracted. An activation switch 1409 may be disposed on the tissue-engaging surface 4 and may be configured to activate the autoinjector 2 or otherwise place the autoinjector 2 into a "ready" mode. A touch sensor 1410 may also be disposed on the tissue-engaging surface 4 and may be configured to assist the controller of the autoinjector 2 in determining whether the autoinjector 2 is placed on the user's skin (indicating that the autoinjector should fire or otherwise deploy the needle) or whether the activation switch 1409 has been improperly triggered (indicating that operation of the autoinjector 2 should cease). A connection port 13 may also be disposed on the tissue-engaging surface 4 to facilitate programming of the autoinjector 2.

[0072] The autoinjector 2 can be configured to operate in three or more operational phases, including, for example, an injection sequence activation phase, an injection phase, and a retraction phase, each of which will be described in further detail herein. The injection sequence activation phase, injection phase, and retraction phase may be collectively referred to herein as the "injection sequence".

[0073] Reference is shown in the cross-section of the autoinjector 2. Figure 3A , Figure 3B and Figure 3C The activation switch 1409 may be a mechanical plunger-type switch. For example, the activation switch 1409 may include a plunger 1450 having a plunger contact surface 1452. The plunger contact surface 1452 may be generally circular in shape (or have another suitable shape) and may be large enough to be comfortably pressed by soft skin. In some embodiments, the plunger contact surface 1452 may have a diameter or width ranging from about 2 mm to about 10 mm, a diameter ranging from about 4 mm to about 8 mm, or a diameter of about 6 mm. The activation switch 1409 may also include a shaft 1442, a biasing member 1444, a biasing collar 1446, and a plunger flange 1454. The biasing member 1444 may be, for example, a spring and may surround the shaft 1442. The biasing member 1444 may be fixed at one end by the biasing collar 1446 or otherwise prevented from moving. The plunger flange 1454 may be configured to contact or otherwise press the plunger switch 1448. For clarity, as used herein, the term activation switch and / or reference to activation switch 1409 should be understood to cover any or all components of activation switch 1409, including shaft 1442, bias member 1444, bias collar 1446, plunger switch 1448, plunger 1450, plunger contact surface 1452, and plunger flange 1454.

[0074] In its free state, that is, when the plunger 1450 is not pressed, whether by pressing against the user's skin or otherwise, the plunger 1450 can extend outward from the tissue engagement surface 4, such as... Figure 3A As shown. In the free state, the distance between the plunger contact surface 1452 and the tissue engagement surface 4 can be in the range of about 1 mm to about 16 mm, about 5 mm to about 12 mm, or about 8.5 mm. In the free state, the biasing member 1444 can cause the plunger 1450 to extend outward from the tissue engagement surface by pressing against the biasing collar 1446. In the free state, the plunger flange 1454 can contact or otherwise press the plunger switch 1448. When the plunger flange 1454 contacts or otherwise presses the plunger switch 1448, the circuit associated with the plunger switch 1448 can be complete or closed.

[0075] When plunger 1450 is pressed by pressing against the user's skin or otherwise, plunger 1450 may initially move to a partially pressed state, such as Figure 3B As shown. In the partially pressed state, the biasing member 1444 can be compressed against the biasing collar 1446. The plunger flange 1454 can also disengage from the plunger switch 1448, or otherwise not be pressed. When the plunger flange 1454 is spaced apart from, not in contact with, or otherwise not pressed by the plunger switch 1448, the circuitry associated with the plunger switch 1448 can be incomplete or disconnected. With this configuration, when the plunger 1450 is pressed, such as when the autoinjector 2 is in its package, the autoinjector 2 can be maintained in a reduced power state.

[0076] like Figure 3B As shown, before the plunger flange 1454 disengages from the plunger switch 1448, the plunger 1450 may not necessarily travel to the fully pressed state (e.g., Figure 3C (As shown). On the other hand, as... Figure 3C As shown, in the fully pressed state, the plunger 1450 can be pressed inward so that the plunger contact surface 1452 is flush or almost flush with the tissue engagement surface 4.

[0077] The plunger flange 1454 can disengage from the plunger switch 1448, for example, after the plunger 1450 has traveled less than 5 mm, less than 3 mm, less than 1 mm, or approximately 0.75 mm—all when, for example, the maximum press distance is 8.5 mm. In other words, after the plunger 1450 has moved only a portion of its maximum press distance relative to the housing 3 of the autoinjector 2, the plunger 1450 can transition from a free state where the plunger flange 1454 is in contact with the plunger switch 1448 to a partially pressed state where the plunger flange 1454 is out of contact with the plunger switch 1448. For example, the plunger 1450 can transition to the pressed state after only moving approximately 5%, approximately 10%, or approximately 20% of the maximum press distance. Therefore, the autoinjector 2 and the plunger switch 1448 can be sufficiently responsive when the plunger 1450 is pressed against the user's skin. For example, the autoinjector 2 and plunger switch 1448 can respond adequately when pressed against skin of varying hardness or by users with different body fat percentages. While examples of the travel distance of the plunger 1450 are provided herein, it should be understood that this disclosure is not limited to any particular example and any suitable travel distance can be used.

[0078] The biasing member 1444 may be sufficiently rigid to allow the plunger flange 1454 to remain in contact with or otherwise continuously press the plunger switch 1448 in the free state. The biasing member 1444 may also have sufficient rigidity to allow the plunger 1450 to be comfortably pressed against the user's skin. The biasing member 1444 may be biased to hold the plunger 1450 in the free state.

[0079] Although the activation switch 1409 is in Figures 3A to 3C The switch 1409 is shown as a mechanical plunger type switch; however, it should be understood that the activation switch 1409 can be any other suitable type of switch, such as a rocker switch, toggle switch, rotary switch, temperature switch, etc. Furthermore, although the circuit associated with plunger switch 1448 is described herein as closed when plunger 1450 is in the free state and open when plunger 1450 is in the pressed state, it should be understood that the reverse configuration can be used. For example, the circuit associated with plunger switch 1448 can be open when plunger 1450 is in the free state and closed when plunger 1450 is in the pressed state.

[0080] The following will refer to Figure 23 The method of controlling the autoinjector 2 based on the positioning of the activation switch 1409 is described in more detail.

[0081] In addition, such as Figure 4B As shown, in some embodiments, the autoinjector 2 may include a plurality of LEDs 52. The LEDs 52 may be arranged in a ring configuration or any other suitable configuration. As described in further detail below, light from one or more LEDs 52 may indicate various operating states of the autoinjector 2.

[0082] Auto-injector housing

[0083] refer to Figure 1A and Figure 1B The housing 3 of the autoinjector 2 may include an upper portion 30. The upper portion 30 may form a portion of the housing 3 opposite to the tissue-contacting surface 4. The upper portion 30 may include a transparent window 50 through which a user can see the contents of the autoinjector 2, including a vial and / or medication contained in the vial. The transparent window 50 may be positioned on a side of the upper portion 30 and may be shaped to conform to the arcuate / curved profile of the upper portion 30, such as... Figure 1 As shown. The transparent window 50 can be a roughly rectangular shape with rounded corners.

[0084] The upper portion 30 may also include a plurality of transparent windows 54. The transparent windows 54 may be formed on the top surface of the upper portion 30 and may be arranged in any suitable configuration, such as a circular, elliptical, rectangular, or linear configuration. For example, the transparent windows 54 may be circumferentially spaced apart from each other. The transparent windows 54 may allow light from one or more LEDs located within the housing 3 to be visible to the user. The light from the one or more LEDs may indicate various operating states of the autoinjector 2, as described herein.

[0085] Transparent windows 50 and 54 can be integrated into the upper portion 30. For example... Figure 1B As shown, the upper portion 30 may include a transparent portion 500 formed of a transparent material. The transparent portion 500 may be continuous, such that the transparent window 50 and the transparent window 54 are formed from a single piece of transparent material. Furthermore, the transparent portion 500 may be integrated into the upper portion 30, such that the upper portion 30 including the transparent window 50 and the transparent window 54 is manufactured as a single component.

[0086] For example, in order to form the upper portion 30 as a single part, the upper portion 30 can be manufactured using a two-color injection molding process. Figure 19 An exemplary method 1900 for injection molding an upper portion 30 using two-color injection molding or insert injection molding is shown. At step 1910, a first material may be deposited into a first mold having a first core and a first cavity. The first material may have low opacity and may be, for example, a transparent material used for transparent windows 50 and 54. The first material may be, for example, clear acrylic, clarified acrylonitrile butadiene styrene (ABS), polycarbonate, polyvinyl chloride (PVC), or polyethylene terephthalate (PETG). The first mold may be configured, for example, to form the transparent portion 500.

[0087] At step 1920, the first core and material in the first mold cavity can move within the second mold cavity to form a second mold. During movement, the first core can hold the first material. The second mold can be configured, for example, to form an upper portion 30. At step 1930, a second material can be deposited into the second mold cavity containing the first material. The second material can be deposited around the first material and the first core in the unoccupied space of the second mold cavity to form the upper portion 30. The second material can be a material with high opacity, such as white plastic. The second material can be, for example, ABS, polycarbonate, ABS-polycarbonate blends, PVC, or PETG.

[0088] Therefore, the generally opaque upper portion 30, including transparent windows 50 and 54, can be formed from two different materials to form a single component. Forming the upper portion 30 as a single component reduces the total number of steps required to assemble the autoinjector 2. For example, in some embodiments, no fastening or bonding steps or materials are required to connect the transparent and opaque portions of the housing. Avoiding unnecessary assembly steps can further improve the aesthetic appearance of the autoinjector 2. Additionally, forming the upper portion 30 as a single component improves the overall structural integrity of the autoinjector 2. Furthermore, forming the upper portion 30 as a single component can reduce or eliminate dents on the aesthetic surface.

[0089] Needle mechanism

[0090] refer to Figures 5 to 11 The needle mechanism 20 includes a bracket 202, which is capable of being positioned in a first position within the housing 3. Figure 6 ) and second positioning ( Figure 7 The needle mechanism 20 may also include a fluid conduit 300 mounted to the bracket 202 and deployable into the user's body, and retractable by the actuator 320. A shuttle 340 (e.g., a shuttle actuator) may be configured to move the actuator 320 via a deployment gear 360 and a retraction gear 362. The shuttle 340 may be coupled to an elastic member (e.g., a spring 370). Cover 380 ( Figure 5 It can be connected to the bracket 202 to enclose various components of the needle mechanism 20.

[0091] refer to Figure 5 The fluid conduit 300 can extend from the first end 302 to the second end 304. For example... Figure 5A As shown in further detail, the first end 302 may include a needle 306 configured to be injected into a user's body. The needle 306 may include a sharp and / or beveled tip and may extend generally along or parallel to axis 44. The second end 304 may include a needle 308 substantially similar to needle 306, but may be positioned within the autoinjector 2 to penetrate cartridge 1302 (in...). Figure 13(As shown in the diagram and described in further detail below) to obtain a drug to be injected into a user. The fluid conduit 300 may include an intermediate segment 310 comprising a portion extending along or parallel to axis 40 and a second portion extending along or parallel to axis 40. The first and second portions of the intermediate segment 310 may be connected in a serpentine segment 312, which facilitates the bending of the fluid conduit 300 and the movement of the needle 306 along axis 44 during deployment into and retraction from the user. While the serpentine segment 312 is shown, any other suitable shape, such as a spiral, a bend, or other shape that allows the fluid conduit 300 to bend, is contemplated. The serpentine segment 312 or similar structure may act as a cantilever when the needle 306 is deployed and / or retracted. The serpentine segment 312 may also bias the fluid conduit 300 to Figure 5 In the deployment configuration shown, once the needle 308 penetrates the cartridge 1302 and establishes fluid communication with it (see example...), Figure 14 The medication can travel from cartridge 1302, through needle 308, intermediate section 310, and needle 306 (piercing the user's skin), and into the user's body. In some instances, the fluid conduit 300 may consist only of metal or a metal alloy. In other instances, the fluid conduit 300 may be any other suitable material, such as a polymer. Needle 308 and intermediate section 310 may define a 22 or 23 gauge thin-walled needle, while needle 306 may be a 27 gauge thin-walled needle. Other needle sizes, ranging from, for example, 6 to 34 gauges, and other needle wall thicknesses, such as standard wall, ultra-thin wall, and extra-thin wall, may also be suitably used. The fluid conduit 300 can reduce the amount of material in contact with the medication, reduce connectors and assembly steps, and require less sterilization than conventional devices.

[0092] like Figure 5B As shown, needle 308 can be configured to include a needle tip 308a and a side port 308b. The side port 308b can be fluidly connected to a fluid path 308c and allows fluid to enter the fluid conduit 300 through the side of needle 308 rather than through the tip of needle 308. The rear wall of the side port 308b can be inclined at an angle θ relative to the longitudinal axis of the fluid path 308c. In some embodiments, the angle θ can be between about 20° and 60°, between about 30° and 50°, or about 40°. By configuring needle 308 in this way, needle 308 can be optimized for piercing the main container of the autoinjector 2, which can be a sealed cartridge or vial. The relative positioning of needle tip 308a and side port 308b allows piercing of the seal of the main container without removing or otherwise cutting a portion of the seal having an opening to the fluid path 308c. Thus, particles removed or cut from the seal can be minimized or avoided from entering the fluid path 308c.

[0093] Needle 306 can be configured to be substantially similar to needle 308, such as Figure 5B As shown. Alternatively, in some embodiments, either or both of needles 306 and 308 may be a 3-bevel needle, a 5-bevel needle, or any other suitable type of needle. In some embodiments, either or both of needles 306 and 308 may be a pencil-tipped needle with a round hole or any other suitable shape of hole.

[0094] The bracket 202 may be formed of plastic (e.g., injection-molded plastic), metal, metal alloy, etc., and may include a flange 204 with an opening 206 and posts 210 and 212. The bracket 202 may also include an opening 216 through which needles or other fluid conduits may be deployed. The opening 216 may be a groove recessed from an end surface of the bracket 202, or, in an alternative embodiment, the entire periphery of the opening 216 may be defined by the material of the bracket 202. The bracket 202 also includes an actuator path 218. The actuator path 218 may be a groove in the bracket 202 extending along or parallel to axis 44. The actuator path 218 may be configured to receive a protrusion of the actuator 320, such as protrusion 330, which is discussed in further detail below. The bracket 202 may also include a shuttle path 220 along which the shuttle 340 may move, as described in further detail below.

[0095] The bracket 202 may also include a stop 240 configured to engage the shuttle 340. The stop 240 may have a fixed end 241. Figure 8 ) and free end 242 ( Figure 8 The cantilever. The stop 240 may include an inclined ramp 243. Figure 9 and Figure 12 When it is on a slope of 1500 (reference) Figure 12 (Description) When engaged or pushed, the stop 240 deflects about the fixed end 241. In a first positioning, the free end 242 may block or otherwise impede movement of the shuttle 340, and in a second configuration, movement of the shuttle 340 may be permitted. The relationship between the stop 240 and the shuttle 340 will be discussed in further detail later in this application.

[0096] The driver 320 includes two racks 322 and 324 that are parallel to each other and disposed on opposite sides of the driver 320. Figure 8 (As shown in the diagram). Racks 322 and 324 may include teeth and may be configured to engage and drive the deployment gear 360 and retraction gear 362 to rotate, respectively. Actuator 320 may include cavity 326 (or track, recess, or other suitable structure). Figure 5 The lumen is configured to receive the needle 306 of the fluid conduit 300. The actuator 320 may also include a protrusion 330.Figure 6 and Figure 7 The protrusion is configured to slide within the drive path 218 of the bracket 202. The protrusion 330 may include a hook-like configuration that can "grab" an obstacle 600, as described in further detail below.

[0097] Continue to refer to Figure 5 The shuttle 340 may include a rack 342 configured to engage with gears 360 and 362. The shuttle 340 may also include an end surface 344 and a recess 346 extending along the length of the shuttle 340 in the same direction as the rack 342. A groove 348 ( Figure 9 The groove 348 may extend along the length of the recess 346. The groove 348 may extend through the middle of the recess 346 and may extend along the entirety or substantially the entirety of the recess 346.

[0098] The shuttle 340 can be positioned along the track 220 from the first starting point. Figure 8 Move to the second intermediate position ( Figure 9 and Figure 10 ), and move from the second location to the third final location (in Figure 11 (Seen between the second and third configurations). As shuttle 340 moves along track 220, rack 342 may first engage deployment gear 360 and then retraction gear 362. At certain times, rack 342 engages at most one of deployment gear 360 and retraction gear 362 at any given time. In some instances, such as when rack 342 is longitudinally positioned between deployment gear 360 and retraction gear 362, rack 342 does not engage either deployment gear 360 or retraction gear 362. Shuttle 340 may be configured to move only along one axis (e.g., axis 40) and only in one direction along that axis. The force required to move shuttle 340 along track 220 may be provided by the expansion of spring 370. Spring 370 can be compressed from a rest state, and the expansion of spring 370 may move shuttle 340 along track 220 through the aforementioned series of positioning / configurations. At various positioning points of the shuttle 340, different features of the autoinjector 2 can directly or indirectly obstruct the movement of the shuttle 340. Alternatively, it is conceivable that the spring 370 can be biased into a compression configuration. In this alternative embodiment, the spring 370 can expand from a resting state, and the compression of the spring 370 can cause the shuttle 340 to move along the track 220 through the aforementioned series of positioning / configurations.

[0099] Figure 8The first positioning of the shuttle 340 shown can correspond to the unused, undeployed, and / or new state of the autoinjector 2. In this first positioning, the actuator 320 can be in an undeployed state. This is achieved by positioning the obstacle 600 in the path of the actuator 320. Figure 6 The obstruction 600 holds the shuttle 340 in a first position. The obstruction 600 may be a shelf of the housing 3 or another suitable blocking device that can prevent movement of the drive 320 by engaging and / or retaining the protrusion 330. Therefore, since the drive 320, deployment gear 360, and rack 342 are interconnected, the obstruction of the drive 320 also prevents movement of the shuttle 340. The shuttle 340 can be moved from the first position to a second position (and vice versa) by moving the obstruction 600 relative to the bracket 202. In one example, the bracket 202 moves (e.g., moves to...) Figure 6 (on the left side of the middle), while obstacle 600 remains stationary.

[0100] When there are no obstacles in the path of driver 320 600 ( Figure 7 When the spring 370 expands, it moves the shuttle 340 along the track 220. This linear movement of the shuttle 340 can cause the deployment gear 360 to rotate counterclockwise (or clockwise in other instances) via the rack 342, and the rotation of the deployment gear 360 can cause the driver 320 to move downward along the axis 44 via the rack 322 of the driver 320. This downward movement of the driver 320 may cause the needle 306 to pierce the user's skin. In some instances, the driver 320 may be configured to move only along the axis 44 relative to the bracket 202.

[0101] The shuttle 340 can be moved by the expansion of the spring 370 until its end surface 344 abuts the free end 242 of the stop 240, thus maintaining the shuttle 340 in position. Figure 9 and Figure 10 The second positioning is shown. In this position, the free end 242 prevents further expansion of the spring 370 and further movement of the shuttle 340 along the track 220. In this second positioning, the fluid conduit 300 can be deployed into the user's body, and fluid from the cartridge 1302 can be injected into the user's body via the needle 306. Additionally, when the shuttle 340 is in the second positioning, the rack 342 can engage with the deployment gear 360 to hold the needle 306 in the deployment configuration. The shuttle 340 can move from the second positioning to the third positioning by bending the stop 240 around its fixed end 241. (Refer to below...) Figures 12 to 14Further details of the bend are provided. The bend of the stop 240 allows the spring 370 to continue expanding, thereby further pushing the shuttle 340 along the track 220. In some instances, the stop 240 may be received by and / or within a recess 346 of the shuttle 340, and the ramp 243 may slide within a groove 348 as the shuttle 340 moves from the second position to the third position.

[0102] The movement of the shuttle 340 from the second position to the third position corresponds to the retraction of the needle 306 from the user into the housing 3. Specifically, the rack 342 can engage with the retraction gear 362 and rotate the retraction gear in the same direction as the rotation of the deployment gear 360 (e.g., counterclockwise or clockwise). The rotation of the retraction gear 362 can cause the actuator 320 to return to the retracted position via the rack 324. The shuttle can reach the third position when the end surface 344 of the shuttle 340 engages the wall of the bracket 202, when the free end 242 of the stop 240 reaches the end of the recess 346, and / or when the spring 370 reaches a stationary state, at which point the actuator 320 is fully retracted.

[0103] In some embodiments, once the actuator 320 moves from the deployed state back to the retracted state, it can be prevented from moving out of the retracted state. Therefore, the needle 306 is prevented from being redeployed into the user's body. In this configuration, the autoinjector 2 can be a disposable device (e.g., discarded after one injection). In other embodiments, the autoinjector 2 can be reset and reused. Furthermore, in some instances, the deployment gear 360 and the retraction gear 362 can be the only rotating gears disposed within the autoinjector 2.

[0104] Piercing system and aseptic connector

[0105] Figure 13 and Figure 14 The puncture system 1300 of an autoinjector 2 is shown. Further details of the exemplary puncture system can be found in U.S. Patent Application Publication No. 2016 / 0262984A1, issued to Arnott et al., published September 15, 2016, the entire contents of which are incorporated herein by reference. The puncture system 1300 includes a main container, which may be a cartridge 1302 having a first end 1304 and a second end 1306. The main container may alternatively be a chamber, syringe, vial, flexible capsule, or any other suitable fluid-containing structure.

[0106] The cartridge 1302 may include a cavity 1308 that opens at a first end 1304 and extends toward a second end 1306. The second end 1306 may include a neck 1310 having a cap 1312 that engages the neck 1310 to close the second end 1306. A diaphragm 1314 may be positioned between the cartridge 1302 and the cap 1312 to help close the second end 1306 and allow a needle 308 (e.g., a post needle) to be inserted into the cartridge 1302. The cavity 1308 may be closed at the first end 1304 by a piston 1316.

[0107] In some instances, cartridge 1302 may have a capacity of 5 mL, but any other suitable volume may be used depending on the drug to be delivered (e.g., 1 mL to 50 mL, or 2 mL to 10 mL, or 3 mL to 6 mL, or 2 mL to 5 mL, or 10 mL to 20 mL, or 10 mL to 30 mL, or another suitable range). In some instances, cartridge 1302 may have a capacity of 10 mL, 15 mL, 20 mL, 25 mL, 30 mL, 35 mL, 40 mL, 45 mL, or 50 mL. In other instances, cartridge 1302 may have a capacity greater than or equal to 1 mL, or greater than or equal to 2 mL, or greater than or equal to 3 mL, or greater than or equal to 4 mL, or greater than or equal to 5 mL, or greater than or equal to 10 mL, or greater than or equal to 15 mL. Cartridge 1302 can contain and preserve drugs for injection into the user and can help maintain the sterility of the drug. The cartridge 1302 may have a neck diameter of 13 mm, a length of 45 mm, and an inner diameter of 19.05 mm. These values ​​are merely exemplary, and other suitable dimensions may be used appropriately. In some instances, the cartridge 1302 may be formed using common materials and may be shorter than existing devices, which helps the autoinjector 2 remain cost-effective and compact. The cartridge 1302 may be a shortened ISO 10 mL cartridge.

[0108] The diaphragm 1314 may comprise an uncoated bromobutyl material or another suitable material. The piston 1316 may comprise a bromobutyl material coated with a fluoropolymer, and may also include a tapered nose 1316a to help reduce dead volume within the cartridge 1302. The piston 1316 may comprise one or more rubber materials, such as halogenated butyl rubber (e.g., bromobutyl rubber, chlorobutyl rubber, fluorobutyl rubber) and / or nitrile and other materials.

[0109] The puncture system 1300 may further include a top 1354 located at the second end 1306. The top 1354 may include a base 1355 positioned above the openings of the diaphragm 1314 and the cartridge 1302. The top 1354 may include a chamber 1356 extending from the base 1355 in a direction away from the piston 1316. The chamber 1356 defines a cavity 1357 and includes an opening 1358 communicating with the cavity 1357. In some embodiments, the top 1354 may be integrated with the diaphragm 1314 (e.g., a one-piece or integral construction). In alternative embodiments (not shown), the top 1354 may be disposed on or initially assembled onto the fluid conduit 300 and may not be directly mounted on / with the cartridge 1302 and / or integrated with the diaphragm 1314.

[0110] A portion of the fluid conduit 300, such as a needle 308, a tube, etc., may extend through the opening 1358 of the chamber 1356 and into the cavity 1357, but not through the base 1355 in a pre-activated state. The opening 1358 may be pre-formed or formed by the needle 308 passing through the chamber 1356. The opening 1358 of the chamber 1356 may form a sterile sliding seal around the needle 308, thereby preventing pathogens or other contaminants from entering the cavity 1357. The needle 308 may move relative to its top 1354 without disrupting the sterile seal therebetween. The cavity 1357 may be sterile or decontaminated, such that the inner surfaces of the cavity 1357 and the needle 308 are sterile. In another embodiment, the cavity 1357 may be sterilized after the needle 308 has been inserted into the cavity 1357 through the opening 1358. In an alternative implementation, a coiled, flexible (e.g., rubber) bellows or bladder-like member (instead of the top 1354) can form a cavity 1357 and allow the cartridge 1302 to translate relative to the needle 308 (and vice versa). After sterilization, the flexible member can also seal around the needle 308 or form the cavity 1354.

[0111] Piston 1316 can be coupled to translation mechanism 1366, which is configured to translate piston 1316 and cartridge 1302 in a direction toward second end 1306. The movement of piston 1316 toward second end 1306 causes piston 1316 to act on the contents (e.g., medication, drug) within cartridge 1302, ultimately transmitting force to second end 1306 of cartridge 1302, thereby causing cartridge 1302 to move along longitudinal axis 40. Translation mechanism 1366 may include a 12mm motor with a five-stage gear reduction (360:1). Translation mechanism 1366 may have spring contacts forming an electrical connection with an associated printed circuit board (e.g., first electronic board 1402). The motor may be configured to generate approximately 136 mN*m of torque at 36 rpm. These design parameters of the motor are merely exemplary, and any other suitable motor may be used.

[0112] The translation mechanism 1366 may include a lead screw mechanism coupled to the piston 1316, which extends axially during relative rotation about a longitudinal axis 40. This telescopic lead screw may have an output of 100 N, a stroke of 20 mm, and a 7° / 45° support thread shape with a pitch of 0.75 mm. Materials used for the lead screw mechanism may include acetal and polybutylene terephthalate. The lead screw mechanism may extend within the piston 1316 to reduce dead space behind the piston 1316. Although the piston 1316 is in Figure 13 and Figure 14 The piston 1316 is shown as having longitudinally spaced threads, but in some instances, such threads may not be present. In another exemplary embodiment (not shown), the translation mechanism 1366 may include a manually engageable surface or component that is manually operated by a user to move the piston 1316. For example, the puncture system 1300 may include a cartridge or plunger coupled to the rear side of the piston 1316. In another exemplary embodiment (not shown), the translation mechanism 1366 may include a pneumatic or hydraulic drive component that is actuated or initiated by a user to move the piston 1316. For example, the drive component may be in the form of an expansion bellows, an expansion bladder, an expansion diaphragm, or a sliding seal or piston. Direct pneumatic or hydraulic pressure can provide the force required to move the piston 1316.

[0113] The puncture system 1300 also includes a collar 1390 coupled to or secured to the second end 1306. The collar 1390 may include a plurality of circumferentially spaced fingers 1392 that engage and surround the neck 1310. The collar 1390 may be secured or otherwise coupled to the second end 1306. The collar 1390 may include a wall 1390a extending at least partially around the neck 1310, the opening of the second end 1306, the cap 1312, the diaphragm 1314, and / or the top 1354. The wall 1390a of the collar 1390 may be located radially or laterally outward of the neck 1310 and extend longitudinally through the neck 1310, the cap 1312, and the diaphragm 1314.

[0114] exist Figure 13 In the pre-activated state of the puncture system 1300 shown, the edge 1393 of the collar 1390 can engage a corresponding radially or laterally inwardly extending cam, latch, or actuating portion 1394 of the actuator retainer member 1395. The retainer member 1395 can slide relative to the collar 1390. The collar 1390 and the retainer member 1395 can be configured such that... Figure 13In the pre-activated state or arrangement shown, at least a portion of the cam or actuation portion 1394 of the retainer member 1395 is positioned directly behind the retaining portion 1399 of the actuator 1398, which is slidable within the retainer member 1395. The wall 1391 of the actuator 1398 may extend into and through the end cap portion 1396 of the retainer member 1395 and into the inner portion of the retainer member 1395, and the retaining portion 1399 of the actuator 1398 may extend radially outward from the wall 1391. In some embodiments, the wall 1391 of the actuator 1398 may be substantially cylindrical, and the retaining portion 1399 of the actuator 1398 may be a flange extending around the end of the wall 1391.

[0115] In the pre-activated state of the puncture system 1300, a biasing or elastic member 1397 capable of elastic deformation can be positioned between the cap portion 1396 of the retainer member 1395 and the retaining portion 1399 of the actuator 1398. The biasing member 1397 can apply a force to the actuator 1398 in the direction toward the cartridge 1302 in the pre-activated state of the puncture system 1300. The biasing member 1397 can be any member that effectively applies force in the pre-activated state and then releases this force upon activation, as described below. Figure 14 As discussed. In some embodiments, the biasing member 1397 may be a tapered or flat spring.

[0116] The needle 308 of the fluid conduit 300 can be fixed or coupled to the actuator 1398, allowing the fluid conduit 300 to move together with the actuator 1398. In the pre-activated state of the puncture system 1300, the needle 308 can be positioned within the sterile cavity 1357, but does not pass through the base 1355 of the top 1354, the diaphragm 1314, and / or enter the cavity 1308 of the cartridge 1302.

[0117] In some embodiments, instead of cavity 1357, needle 308 can be positioned within a stopper when puncture system 1300 is in a pre-activated state. The stopper can be a solid stopper without any holes, cavities, or openings, and can be formed of a first rubber material. The first rubber material can be permeable to sterilizing gases, such as ethylene oxide or vaporized hydrogen peroxide. The first rubber material can include one or more of isoprene, ethylene propylene diene monomer (M grade) rubber (EPDM), and styrene-butadiene. The permeability of the first rubber material to sterilizing gases allows needle 308 to be sterilized before use when positioned within the stopper. The stopper can be molded around needle 308 such that needle 308 is punctured into the stopper.

[0118] In order to make the puncture system 1300 from Figure 13The pre-activated state movement can activate the translation mechanism 1366 to move the piston 1316 toward the second end 1306 and translate the cartridge 1302 along the longitudinal axis 40 toward the actuator 1398. Since the needle 308 is not yet in fluid communication with the cartridge 1302, the activation of the translation mechanism 1366 applies pressure to the fluid contained in the cartridge 1302, and then applies that pressure to the cartridge 1302 itself. This pressure also causes the edge 1393 to push against the actuating portion 1394 and deflect the actuating portion radially outward. Without the actuating portion 1394 obstructing its path, the holding portion 1399 and the needle 308 move toward the cartridge 1302 by the expansion of the biasing member 1397. The actuator 1398 can be coupled to the flange 204 of the bracket 202, and therefore, this movement of the actuator 1398 toward the cartridge 1302 can also move the bracket 202 in the same direction. This movement corresponds to the bracket 202 relative to Figure 6 and Figure 7 The movement of the housing 3 in the middle allows the protrusion 330 to clear the obstacle 600 to inject the needle 306.

[0119] Movement of needle 308 toward the second end 1306 of cartridge 1302 also causes needle 308 to pierce the base 1355 of top 1354, diaphragm 1314, and cavity 1308, establishing fluid communication with the contents of cartridge 1302. Once needle 308 is in fluid communication with cartridge 1302, further movement of piston 1316 toward the second end 1306 pushes fluid through needle 308 and the remainder of fluid conduit 300. In some embodiments, puncture system 1300 may be configured such that, after activation, no portion of needle 308 extends into cavity 1308 beyond the portion already positioned within sterile cavity 1357. This helps prevent non-sterile portions of needle 308 from contaminating the contents of cartridge 1302.

[0120] The biasing member 1397 can be configured to expand such that the fluid conduit 300 pierces the top 1354 and / or the diaphragm 1314 at a high speed (such as at least about 10 mm / s or at least about 40 mm / s). The relatively rapid piercing of the top 1354 and / or the diaphragm 1314 via the biasing member 1397 can help prevent leakage of the contents of the cavity 1308, which can be under pressure via the piston 1316.

[0121] After the medication has been delivered to the user by needle 306, needle 306 can be automatically withdrawn from the user. (Reference) Figures 12 to 14The translation mechanism 1366 can operate in reverse mode, causing the lead screw to rotate in the opposite direction compared to the insertion step. This reverse rotation causes the piston 316 to move rearward toward the first end 1304 and also causes the cartridge 1302 to move in the opposite direction along axis 40 (compared to during fluid delivery and needle 306 insertion). The movement of the cartridge 1302 in the opposite direction allows... Figure 12 The ramp 1500 (attached to wall 1391) pushes against the ramp 243 of the stop 240. This allows the stop 240 to deflect about its fixed end 241 in the direction of arrow 240a and allows the shuttle 340 to move from its second position to its third position to retract the needle 306 as described above. In this way, a single spring within the device can be used to withdraw and insert the needle into the patient.

[0122] It is also conceivable that the fluid conduit 300 could be the only fluid conduit of the autoinjector 2 configured to be in fluid communication with the cartridge 1302. Therefore, during normal operation of the autoinjector 2, medication from the cartridge 1302 can be delivered to the user solely through the fluid conduit 300. Additionally, the needle 306 could be the only needle of the autoinjector 2 configured to be delivered into the patient. In this way, fluid can be transported from the cartridge 1302 to the patient using a single piece of metal or plastic.

[0123] Figures 14A to 14D An exemplary configuration of an external screw 1367 that can be incorporated into a translation mechanism 1366 and can mat with a piston 1316 is depicted. The external screw 1367 typically includes an upper portion 1378, a lower portion 1381, and a flange portion 1375. The flange portion 1375 may have a first surface 1376 and a second surface 1377, and separates the upper portion 1378 and the lower portion 1381 of the external screw 1367. A conduit 1372 extends the length of the external screw 1367 and typically has a uniform diameter.

[0124] The upper portion 1378 may typically include one or more protrusions 1379 projecting from the first surface 1376 of the flange portion 1375. In some embodiments, the one or more protrusions 1379 may be formed or partially formed by an ejector pin during the molding process and are typically configured to fit into a corresponding feature of the output gear of the motor assembly. It should be understood that any number of one or more protrusions 1379 may be present, and the one or more protrusions 1379 may have any suitable shape and / or size. In some embodiments, at least two, four, or five protrusions are present.

[0125] The lower portion 1381 typically includes a nose region 1382 and one or more protrusions 1383 extending radially outward from the outer surface 1373 of the conduit 1372 and longitudinally extending from the second surface 1377 of the flange portion 1375. Each of the one or more protrusions 1383 typically includes a tip 1384 and a surrounding side edge 1385. The one or more protrusions 1383 are configured to interlock with corresponding features on the interior of the piston 1316. The side edge 1385 has a chamfered design, and the tip is typically tapered to facilitate the insertion of an external screw 1367 into the piston 1316 during assembly. The nose region 1382 extends from the tip 1384 of the one or more protrusions 1383 to the end of the lower portion 1381. The nose region 1382 may include a chamfer toward the end of the lower portion 1381 to further facilitate the insertion of the external screw 1367 into the piston 1316.

[0126] The configuration of the external screw 1367 (including its dimensions and tolerances) can be selected to balance ease of assembly, strength and durability of the coupling mechanism, and the degree of deformation of the piston 1316 under external pressure. For example, the nose region 1382 can be configured to facilitate the insertion of the external screw 1367 into the piston 1316. Furthermore, the shape and size of the tip 1384 can be designed to facilitate mating with a feature of the piston 1316. For example, the tip 1384 can typically be a triangular shape with smooth edges, which facilitates the advancement of the protrusion 1383 through the internal feature of the piston 1316. By facilitating the insertion of the external screw 1367 into the piston 1316 during assembly, dislodgement of the piston 1316 from its position within the cartridge 1302 can be prevented, and the integrity of the seal of the cartridge 1302 can be maintained during assembly.

[0127] The length L1 of the external screw 1367 can range from about 12 mm to about 16 mm, including all subranges and values ​​therein. In some embodiments, the length L1 of the external screw 1367 can range from about 12.5 mm to about 15.5 mm, about 12.75 mm to about 15.25 mm, about 13 mm to about 15 mm, or about 14 mm to about 15 mm. In some embodiments, the length L1 of the external screw 1367 can be about 14.42 mm, about 14.48 mm, about 14.54 mm, about 14.60 mm, about 14.66 mm, about 14.72 mm, about 14.78 mm, about 14.84 mm, about 14.90 mm, or about 14.96 mm.

[0128] The length L2 of the lower portion 1381 of the external screw 1367 can be in the range of about 8.0 mm to about 13.0 mm, including all subranges and values ​​therein. In some embodiments, the length L2 of the lower portion 1381 of the external screw 1367 can be in the range of about 8.0 mm to about 12.0 mm, about 8.0 mm to about 11 mm, about 8.0 mm to about 10.5 mm, about 9.0 mm to about 12.0 mm, about 9.0 mm to about 11.0 mm, about 9.0 mm to about 10.5 mm, about 9.5 mm to about 10.5 mm, or about 9.9 mm to about 10.2 mm. In some embodiments, the length L2 of the lower portion 1381 of the external screw 1367 may be approximately 9.62 mm, approximately 9.68 mm, approximately 9.74 mm, approximately 9.80 mm, approximately 9.86 mm, approximately 9.92 mm, approximately 9.98 mm, approximately 10.04 mm, approximately 10.10 mm, approximately 10.16 mm, approximately 10.22 mm, approximately 10.28 mm, approximately 10.34 mm, or approximately 10.40 mm.

[0129] The length L3 of the nose region 1382 of the lower portion 1381 of the external screw 1367 can range from about 0.5 mm to about 7.0 mm, including all subranges and values ​​therein. In some embodiments, the length L3 of the nose region 1382 of the lower portion 1381 of the external screw 1367 can range from about 1.0 mm to about 4.5 mm, about 1.5 mm to about 4.0 mm, about 1.5 mm to about 3.5 mm, about 2.0 mm to about 5.0 mm, about 2.5 mm to about 5.0 mm, or about 3.0 mm to about 5.0 mm. In some embodiments, the length L3 of the nose region 1382 of the lower portion 1381 of the external screw 1367 can be about 5.41 mm, about 5.47 mm, about 5.53 mm, about 5.59 mm, about 5.65 mm, about 5.71 mm, about 5.77 mm, about 5.83 mm, about 5.89 mm, about 5.95 mm, about 6.01 mm, about 6.07 mm, about 6.13 mm, about 6.19 mm, about 6.25 mm, about 6.31 mm, or about 6.37 mm. The tolerances for the lengths described herein can be about 0.2 mm or less, about 0.15 mm or less, about 0.12 mm or less, about 0.10 mm or less, about 0.08 mm or less, or about 0.06 mm or less.

[0130] The threaded region 1380 of the conduit 1372 can typically be characterized by several diameter measurements. For example, the threaded region 1380 can be defined by a small diameter measured between two opposing peaks of the thread, a large diameter measured between two opposing roots of the thread, or a pitch circle diameter measured between the midpoints of the thread on either side of the threaded region 1380. In some embodiments, the threaded region 1380 of the conduit 1372 may have a small diameter in the range of about 6.75 mm to about 7.60 mm, a large diameter in the range of about 7.75 mm to about 8.40 mm, and / or a pitch circle diameter in the range of about 7.00 mm to 8.15 mm, including all subranges and values ​​therebetween.

[0131] The small diameter can be in the range of about 6.75 mm to about 7.50 mm, about 6.75 mm to about 7.40 mm, about 6.75 mm to about 7.30 mm, about 7.00 mm to about 7.50 mm, about 7.00 mm to about 7.40 mm, about 7.00 mm to about 7.30 mm, about 7.15 mm to about 7.50 mm, about 7.15 mm to about 7.40 mm, or about 7.15 mm to about 7.30 mm. The large diameter can be in the range of approximately 7.75 mm to approximately 8.30 mm, approximately 7.75 mm to approximately 8.20 mm, approximately 7.75 mm to approximately 7.10 mm, approximately 7.90 mm to approximately 8.30 mm, approximately 7.90 mm to approximately 8.20 mm, approximately 7.90 mm to approximately 8.10 mm, approximately 8.05 mm to approximately 8.30 mm, approximately 8.05 mm to approximately 8.25 mm, approximately 8.05 mm to approximately 8.20 mm, approximately 8.05 mm to approximately 8.15 mm, or approximately 8.08 mm to approximately 8.15 mm. The pitch circle diameter can be in the range of about 7.00 mm to about 8.10 mm, about 7.25 mm to about 8.00 mm, about 7.25 mm to about 7.90 mm, about 7.25 mm to about 7.75 mm, about 7.25 mm to about 7.60 mm, about 7.45 mm to about 8.10 mm, about 7.45 mm to about 8.00 mm, about 7.45 mm to about 7.90 mm, about 7.45 mm to about 7.75 mm, about 7.45 mm to about 7.60 mm, or about 7.50 mm to about 7.60 mm. In some embodiments, the smaller diameter may be about 6.93 mm, about 7.00 mm, about 7.07 mm, about 7.14 mm, about 7.21 mm, about 7.28 mm, about 7.35 mm, about 7.42 mm, or about 7.49 mm; the larger diameter may be about 7.78 mm, about 7.86 mm, about 7.94 mm, about 8.02 mm, about 8.10 mm, about 8.18 mm, about 8.26 mm, or about 8.34 mm; and the pitch circle diameter may be about 7.20 mm, about 7.26 mm, about 7.32 mm, about 7.38 mm, about 7.44 mm, about 7.50 mm, about 7.56 mm, about 7.62 mm, about 7.68 mm, about 7.74 mm, about 7.80 mm, or about 7.86 mm.

[0132] Locking member (latch)

[0133] refer to Figures 16A to 16D The auto-injector 2 may include a locking component 1610. For example... Figure 16A As shown, the locking component 1610 may include a lock (e.g., a protrusion) 1612 having a curved surface 1614, and may also include a cover portion 1616. The shape of the cover portion 1616 may be designed to conform to the tissue engagement surface 4 of the autoinjector 2, such as...Figure 16D As shown. The cover portion 1616 and the tissue engagement surface 4 may be concave to receive the user's anatomical portion 1600. The anatomical portion 1600 may be, for example, the thigh, buttock, arm, back, or any other area of ​​the body suitable for injection. Lock 1612 may be attached to the cover portion 1616. In some embodiments, the locking member 1610 may be formed as a single piece, such that the lock 1612 and the cover portion 1616 are integrally connected. The locking member 1610 may be formed of any suitable rigid or semi-rigid material. The locking member 1610 may be formed, for example, of acrylonitrile butadiene styrene (ABS), and may also have a frosted clear appearance indicating that the locking member 1610 is disposable.

[0134] like Figures 16B to 16C As shown, the locking member 1610 may be disposed on or adjacent to the tissue engagement surface 4 of the autoinjector 2, such that the lock 1612 may extend into the autoinjector 2. The locking member 1610 may also be disposed on or adjacent to the liner 12a, which may initially cover the adhesive patch 12 before use of the autoinjector 2. The locking member 1610 may be positioned relative to the liner 12a such that when the liner 12a is removed from the adhesive patch 12, the locking member 1610 may also be removed from the tissue engagement surface 4. The lock 1612 may extend into the autoinjector 2 via a locking opening 1630 formed in the tissue engagement surface 4. When the locking member 1610 is disposed on or adjacent to the tissue engagement surface 4, the cover portion 1616 may be attached to the tissue engagement surface 4 via an adhesive disposed between the cover portion 1616 and the tissue engagement surface 4. The locking member 1610 may be disposed on or adjacent to the tissue engagement surface 4 such that it can be selectively removed by the user.

[0135] refer to Figures 16C to 16D When the locking member 1610 is disposed on or near the tissue engagement surface 4, the lock 1612 may extend into the autoinjector 2, thereby preventing movement of one or more internal mechanisms of the autoinjector 2. For example, when the locking member 1610 is disposed on or near the autoinjector 2, the lock 1612 may extend into the autoinjector 2, thereby engaging with one or more internal components of the autoinjector 2 to prevent movement and / or activation of these components.

[0136] refer to Figure 16DWhen the locking member 1610 is positioned on or near the tissue engagement surface 4, the lock 1612 can extend into the autoinjector 2, thereby positioning it within the puncture system 1300. As previously described herein, the collar 1390 can be coupled or secured to the second end 1306 of the cartridge 1302. Also as previously described herein, when the puncture system 1300 moves from a pre-activated state, the cartridge 1302, and therefore the collar 1390, can translate toward the holding portion 1399 in a direction parallel to the longitudinal axis of the cartridge 1302. When the lock 1612 extends into the autoinjector 2 and is adjacent to the collar 1390, the lock 1612 can prevent the cartridge 1302 from translating toward the holding portion 1399 or otherwise prevent the cartridge 1302 and the collar 1390 from exerting force on the actuating portion 1394. Therefore, even if the motor is activated in some way when the lock 1612 is in its locked position, fluid communication between the needle 308 and the cartridge 1302 cannot be established, and the needle 306 cannot be deployed outside the housing 3. Furthermore, when in the locked position, the lock 1612 prevents the cartridge 1302 from moving toward the needle 308, thereby preventing the actuation portion 1394 from deflecting and thus preventing the holding portion 1399 and the needle 308 from moving toward the cartridge 1302. In the event of the autoinjector 2 being dropped or subjected to vibration, the lock 1612 further prevents the puncture system 1300 from moving from the pre-activated state, and thus prevents the needle 308 from puncturing the cartridge 1302.

[0137] When the locking member 1610 is disposed on or adjacent to the tissue engagement surface 4, the locking member 1610 can also serve as a spacer between the user's skin and the tissue engagement surface 4. For example, the locking member 1610 can have a thickness such that the touch sensor 1410, described in more detail below, cannot detect the user's skin, thereby preventing accidental activation of the autoinjector 2. The locking member 1610 can have a thickness of, for example, from about 1 mm to about 5 mm or about 3 mm.

[0138] Therefore, the locking member 1610 can act as an effective safety mechanism to prevent accidental activation of the autoinjector 2. When the locking member 1610 is positioned on or near the tissue engagement surface 4, the lock 1612 can prevent movement of various internal components of the autoinjector 2. For example, in the event that the autoinjector 2 is dropped on the floor before use, the locking member 1610 can prevent accidental puncture of the cartridge 1302 and / or accidental activation of the injection sequence. The locking member 1610 can also prevent such movement and / or accidental activation of the injection sequence when the autoinjector 2 is subjected to vibration during transport.

[0139] If the user wishes to use and / or is ready to use the autoinjector 2, the user can separate the locking member 1610 from the tissue engagement surface 4, thereby removing the lock 1612 from the lock opening 1630. The user can, for example, peel the covering portion 1616 from the tissue engagement surface 4. Alternatively, the user can peel the pad 12a from the adhesive patch 12, thereby removing the locking member 1610 from the tissue engagement surface 4. When the locking member 1610 is separated from the tissue engagement surface 4, the curved surface 1614 allows the lock 1612 to rock within the lock opening 1630, thereby allowing the lock 1612 to be easily removed from the lock opening 1630. With the lock 1612 removed from the lock opening 1630, the autoinjector 2 can be in its ready-to-use state, allowing, for example, an injection sequence to be initiated.

[0140] Figure 16E and Figure 16F A locking component 1610 according to some embodiments is depicted. For example... Figure 16E and Figure 16F As shown, the locking member 1610 may have an increased width (relative to) Figures 16A to 16C (The depiction of the locking member 1610 in the image) ensures that the locking member 1610 extends above the touch sensor 1410 when it is positioned on the autoinjector 2. Furthermore, the locking member 1610 may have a ribbed structure and may include an air gap or recess 1618 and a hinge 1620. When the autoinjector 2 is placed near the user and the locking member 1610 is in place, the air gap 1618 can suppress the conduction of the capacitive field between the user's skin and the touch sensor 1410. When the locking member 1610 is detached from the autoinjector 2, the hinge 1620 can allow the locking member 1610 to bend. In some embodiments, a removable cover, other than and / or separate from the locking member 1610, may extend above the touch sensor 1410 to suppress accidental detection of skin.

[0141] Electronics

[0142] Figure 4A A control system 1400 for an auto-injector 2 is shown. The control system 1400 may include components positioned on a first electronic board 1402 and a second electronic board 1404, and may also include a power supply 1406. The first electronic board 1402 may include a controller 1408, an activation switch 1409, a touch sensor 1410, a needle insertion switch 1412, and a transmitter 1414. The second electronic board 1404 may include a detector 1416, an audio module 1418, a vision module 1420, and a tactile module 1422. Although... Figure 4AAn audio module 1418, a vision module 1420, and a tactile module 1422 are depicted in a second electronic board 1404; however, in some embodiments, one or more of these modules may be included on a first electronic board 1402. One or more components of the first electronic board 1402 and the second electronic board 1404 may be operatively coupled to a controller 1408 and powered by a power supply 1406. The controller 1408 may also be operatively coupled to a translation mechanism 1366 and may be configured to control the operation of the translation mechanism 1366 to initiate and control needle insertion and retraction as described above. During the final assembly step of inserting the cartridge 1302 into the housing 3, the translation mechanism 1366 may be coupled to the first electronic board 1402 via one or more spring contacts. As previously described herein, the translation mechanism 1366 may include a motor, a transmission, and a lead screw mechanism.

[0143] Most of the assembly of the autoinjector 2 can be performed, for example, on an assembly line at a manufacturing plant. The two device halves (or parts) can then be shipped to a drug filling or final assembly plant. In practice, the two separate parts 1490 and 1492 do not need to have the same dimensions, as... Figure 4B As shown. Once the vial (e.g., cartridge 1302) is filled with medication or other drugs, cartridge 1302 can be assembled with the rest of the autoinjector 2. For example, two device halves (parts 1490 and 1492) can be assembled with the cartridge 1302 filled therein. In one example, part 1490 and translation mechanism 1366 can snap into place at the rear of cartridge 1302. Part 1490 can be part of housing 3, which includes a base or module configured to contain translation mechanism 1366 and its associated electronics. Part 1492 can be part of housing 3, which includes substantially all other components described herein, including, for example, the needle mechanism, sterile connector, and puncture mechanism described herein. In this example, the electrical connection of the motor of the translation mechanism 1366 must be made while the translation mechanism 1366 is snapped behind the cartridge 1302 (i.e., during the assembly step of combining parts 1490 and 1492 and cartridge 1302 to form a complete and fully functional autoinjector 2). To accommodate this electrical connection, the drive mechanism of the translation mechanism 1366 may include one or more spring contacts 1494 (see...). Figure 4C These spring contacts will contact pads 1495 on the first electronic board 1402 during assembly (see also...). Figure 4C Although not in Figure 4CAs depicted, however, the transmission system of the translation mechanism 1366 may include additional spring contacts that can engage additional pads on the first electronic board 1402 during assembly. Such additional spring contacts and additional pads can be used to connect additional components of the translation mechanism 1366 (such as a tachometer, motor encoder, or any other sensor or device) to the first electronic board 1402. Therefore, the connection from the translation mechanism 1366 to the first electronic board 1402 (including the controller 1408) can be made without any loose wires or other similar structures.

[0144] Such an assembly process can be relatively simpler than that of simpler devices (e.g., autoinjectors) with relatively more complex final assembly processes. Therefore, the proposed assembly process described herein can reduce labor costs.

[0145] In some implementations, the auto-injector 2 may include, for example: Figure 17 and Figure 17A The single (i.e., only or exactly one) electronic board 1710 shown herein may have components of the control system 1400 previously described herein positioned on it. Figure 17A As shown, the electronic board 1710 may include a first segment 1712 and a second segment 1714. The first segment 1712 and the second segment 1714 may be physically and electrically connected via a flexible segment 1716. The flexible segment 1716 may be, for example, a ribbon cable, a flexible conductive substrate, etc. In some embodiments, the flexible segment 1716 may be formed from a glass fiber sheet machined thin enough to bend, and is sometimes referred to as "semi-bent". In some embodiments, the flexible segment 1716 may be formed from a flexible polymer. The flexible polymer may be formed by a process sometimes referred to as "rigid-flexible bonding", in which a sandwich structure of a first portion of glass fiber, a flexible polymer, and a second portion of glass fiber is first formed. The first and second portions of glass fiber may then be removed to leave a thin flexible polymer portion.

[0146] The electronic plate 1710 may include one or more supports 1720 for mounting or otherwise securing the electronic plate 1710 to the interior of the autoinjector 2. The first plate segment 1712 may also include a notch 1718. The notch 1718 may be positioned such that the first plate segment 1712 may be positioned to allow a needle to pass through the notch 1718 during deployment.

[0147] In some embodiments, the first plate segment 1712 may correspond to the first electronic plate 1402, and the second plate segment 1714 may similarly correspond to the second electronic plate 1404, each as previously described herein. By connecting the first plate segment 1712 and the second plate segment 1714 via a flexible segment 1716, the first plate segment 1712 may be positioned adjacent to the tissue engagement surface 4 of the autoinjector 2, while the second plate segment 1714 may be positioned on the opposite side of the autoinjector 2 toward the upper portion 30 of the housing 3. Thus, a single electronic plate 1710 can be used to connect components positioned toward the tissue engagement surface 4 and components positioned toward the upper portion 30. This configuration facilitates the assembly of the autoinjector 2 by avoiding the need for complex wiring or soldering.

[0148] like Figure 17 As shown, the electronic plate 1710 can be positioned within the housing 3. The first plate segment 1712 can be positioned adjacent to the tissue-contacting surface 4, while the second plate segment 1714 can be positioned on the opposite side of the autoinjector 2 (e.g., in...). Figure 17 (Behind the first plate segment 1712). The flexible segment 1716 can be bent or folded to maintain the connection between the first plate segment 1712 and the second plate segment 1714 in such a positioning.

[0149] Touch sensor 1410 may be incorporated in or on a first segment 1712 of electronic board 1710. To allow for adequate detection of the user's skin, touch sensor 1410 and the first segment 1712 may be located near the tissue-bonding surface 4 of housing 3. The tissue-bonding surface 4 of housing 3 or a portion adjacent to touch sensor 1410 may be thin enough to create a detectable electric field between touch sensor 1410 and the user's skin. In some embodiments, the portion of tissue-bonding surface 4 adjacent to touch sensor 1410 may be less than about 2 mm, about 1 mm, or less than about 1 mm. Furthermore, the portion of tissue-bonding surface 4 adjacent to touch sensor 1410 may be made of a solid material (such as plastic). By forming the portion of tissue-bonding surface 4 adjacent to touch sensor 1410 with a solid material (rather than a ribbed, core-shaped, or hollow material), the dielectric constant between the user's skin and touch sensor 1410 can optimize the responsiveness of touch sensor 1410.

[0150] Additionally, the touch sensor 1410 can be positioned within or on the electronic plate 1710 so as to be adjacent to or near the opening 6 through which the needle can be deployed. By positioning the touch sensor 1410 adjacent to or near the opening 6, the likelihood that the touch sensor 1410 can detect the user's skin when the autoinjector is properly positioned is increased. Furthermore, the curvature of the tissue-attaching surface 4 can reduce the likelihood that the touch sensor 1410 may misinterpret a flat surface, such as a tabletop, as the user's skin by creating space between the touch sensor 1410 and the flat surface.

[0151] By integrating the touch sensor 1410 into or on the electronic board 1710, the need for one or more wires and / or other circuitry to connect the touch sensor 1410 to a separate electronic board can be eliminated. This simplifies the assembly of the autoinjector 2 and reduces the cost of the autoinjector.

[0152] Because the electronic plate 1710 can be positioned adjacent to the tissue bonding surface 4, the electronic plate 1710 may include a notch to allow a needle to pass through the electronic plate 1710 and subsequently through the opening 6 for deployment. Furthermore, the electronic plate 1710 may be positioned such that the touch sensor 1410 is directly adjacent to the opening 6, and there is no gap between the edge of the touch sensor 1410 and the opening 6. Alternatively, the electronic plate 1710 may be positioned such that there is a gap between the edge of the touch sensor 1410 and the opening 6, and this gap has a maximum width of, for example, 5 mm, 2 mm, or 1 mm.

[0153] The controller 1408 can be configured to receive information from the aforementioned system and system components, and process the information according to various algorithms to generate control signals for controlling the internal mechanisms (including the translation mechanism 1366) of the autoinjector 2. See below for reference. Figure 18 and Figures 20 to 23 Describe an example of such an algorithm. The processor can receive information from the system and system components, process the information according to various algorithms, and generate information signals that can be directed to the audio module 1418, vision module 1420, tactile module 1422, or other indicators such as the second electronic board 1404, to inform the user of system status, component status, process status, or any other useful information monitored by the system. The processor can be a digital IC processor, an analog processor, or any other suitable logic or control system that executes control algorithms.

[0154] As mentioned above Figure 3A and Figure 3BThe activation switch 1409 discussed may be a mechanical plunger-type switch extending away from the tissue-contact surface 4 of the autoinjector 2. The activation switch 1409 may include circuitry that remains intact unless the activation switch 1409 is pressed. For example, when the autoinjector 2 is attached to the user's skin, the switch 1409 may be pressed to disconnect the circuitry and instruct the controller 1408 that the autoinjector 2 should be activated. To conserve power, components of the autoinjector 2 may be in an idle or sleep mode until the switch 1409 is activated. In yet another example, the autoinjector 2 may not be powered at all until the switch 1409 is activated, and deactivation of the switch 1409 may completely cut off power to the autoinjector 2. While a mechanical plunger-type switch is disclosed, any other suitable mechanism for activating the autoinjector 2 may be used, including, for example, a button pressed by the user, a voice signal, and a wireless signal from another electronic device.

[0155] Touch sensor 1410 can be configured to help controller 1408 determine whether the autoinjector 2 is correctly deployed on the user's skin. In one example, touch sensor 1410 can be a capacitive sensing electrode or any other device configured to distinguish skin from contact with other materials, such as, for example, wood, plastic, metal, or another material. When skin is near the capacitive sensing electrode, a signal indicating such contact can be sent to controller 1408. Therefore, even when switch 1409 is pressed, touch sensor 1410 can be used to verify that the autoinjector 2 is correctly placed on the user's skin. Touch sensor 1410 may include a capacitive sensing electrode coupled to the first electronic board 1402 and also coupled to the interior of housing 3. Housing 3 and adhesive patch 12 can act as a cover layer (insulator) that serves as a dielectric between the user's skin and the capacitive sensing electrode. Alternatively, the touch sensor 1410 may be incorporated in or on the electronic board 1710, as previously described herein, such that the capacitive sensing electrode is also incorporated in or on the electronic board 1710, and contact between the portion of the housing 3 and / or the adhesive patch 12 near the capacitive sensing electrode may increase the capacitance of the electrode, for example, by about 1 to about 10 pF, thereby indicating that the autoinjector 2 is placed on the skin surface.

[0156] The pin insertion switch 1412 can be configured to send a signal to the controller 1408 that the pin 306 is deployed inside the user's body. For example, refer to... Figure 15The needle insertion switch 1412 may include a curved cantilever 1510, which includes a first contact 1512. The needle insertion switch 1412 may also include a second contact 1514. When the needle 306 is deployed into the user's body, the first contact 1512 may be positioned to make electrical contact with the second contact 1514. During the deployment of the needle 306, the actuator 320 may move downward along axis 44 and deflect the curved cantilever 1510 and the first contact 1512 toward the second contact 1514. When the first contact 1512 and the second contact 1514 are engaged with each other, a signal indicating that the needle 306 has been successfully deployed into the user's body may be sent to the controller 1408. Separation of the first contact 1512 and the second contact 1514 may indicate that the needle 306 has been retracted from the user.

[0157] The transmitter 1414 and detector 1416 can operate as a light interruption sensor or light interruptor to allow the controller 1408 to determine the state of the auto-injector 2. The transmitter 1414 can be a light-emitting diode (LED) or other suitable light emitter, and the detector 1416 can be, for example, a phototransistor configured to receive light emitted by the transmitter 1414. In one example, the transmitter 1414 can emit infrared light, but other suitable wavelengths can also be used. Using infrared light can help reduce interference from external light.

[0158] like Figure 13B As shown, the transmitter 1414 and detector 1416 can be arranged opposite each other within the housing 3 such that the light beam 1430 can pass from the transmitter 1414 through the cartridge 1302 to the detector 1416. The cartridge 1302 and any fluid contained therein can be at least partially transparent to the light beam 1430, allowing the light beam 1430 to pass through the cartridge 1302 and its contents. When the piston 1316 moves toward the second end 1306 during drug delivery (see... Figure 13 and Figure 14Piston 1316 (specifically, the shoulder of piston 1316) can interrupt beam 1430. When detector 1416 fails to sense beam 1430, a signal can be sent to controller 1408, which can interpret the signal to indicate the end of injection (e.g., all medication contained in cartridge 1302 has been expelled). In some instances, the refraction path of beam 1430 can be considered when transmitter 1414 and detector 1416 are positioned relative to each other. For example, beam 1430 can be refracted as it passes through cartridge 1302 and any liquid contained therein, and transmitter 1414 and detector 1416 can be offset from each other accordingly. Alternatively, transmitter 1414 and detector 1416 can be offset from the center of housing 3 such that the shoulder of piston 1316 can block beam 1430. In at least some instances, an optical interruption sensor or similar mechanism can help avoid false alarms in the event of a failure in the drivetrain. That is, compared to other mechanisms, an optical switch can help controller 1408 determine with greater accuracy that injection has not been completed.

[0159] The audio module 1418 may include a speaker or similar device to provide audio feedback to the user. Openings on the housing 3 may facilitate the propagation of sound from the audio module 1418 to the user. For example, the audio module 1418 may generate tones or other sounds at the start and end of the injection, and / or indicate any other benchmarks during the injection, such as errors. The vision module 1420 may include one or more LEDs or similar devices to provide visual feedback to the user. The vision module 1420 may include LEDs of different colors to provide various messages to the user. For example, multiple blue LEDs arranged in a ring may be used to display the progress of the injection over time, one or more green LEDs may be used to indicate the completion of the injection, and red LEDs may be used to indicate errors to the user. In various instances, any other suitable color, combination, and / or number of LEDs may be used. For example, a combination of red, blue, and purple LEDs may be used. In one arrangement, eight LEDs may be arranged in a circle with a diameter of about 26.5 mm or a diameter of about 10.0 mm to about 40.0 mm. It should be understood that this exemplary number and positioning of LEDs is not intended to be limiting, and any number and / or positioning of LEDs may be used. LEDs can be activated sequentially around a circle to indicate the progress of the injection (e.g., a progress ring arranged in a clock-like manner—see example...). Figure 4B(LED 52 on the device). The controller 1408 can also be configured to receive feedback from various sensors and, based on the feedback from the sensors, readjust the activation rate of the various LEDs. For example, LEDs in the progress loop can be activated in three or more operation phases, including, for example, an injection sequence activation phase, an injection phase, and a retraction phase. Those skilled in the art will recognize that the autoinjector 2 can have more or fewer than the three operation phases described above. There may be an expected time to complete each phase, but the actual time experienced during any of the aforementioned operation phases of the autoinjector 2 may also vary. Algorithms can be used to help avoid premature LED activation, for example, when a phase ends earlier than expected, or to stop the progress along the loop when a phase takes longer than expected. At any given point, the algorithm can divide the estimated remaining time for drug delivery completion by the number of inactive LEDs in the progress loop to determine the rate at which the remaining LEDs in the progress loop should be activated.

[0160] For example, prior to the injection sequence activation phase, LEDs can be activated at a rate equal to the estimated time for the entire drug delivery process (e.g., the estimated time to complete all injection sequence activation, injection, and retraction phases) divided by the total number of inactive LEDs in the progress ring. In other words, the estimated time for the entire drug delivery process can be divided by the total number of LEDs in the progress ring minus the number of any activated LEDs. Therefore, for example, if one LED has been activated, the estimated time for the entire drug delivery process can be divided by the total number of LEDs in the progress ring minus one.

[0161] After the injection sequence activation phase is completed, the LEDs can be activated at a rate equal to the sum of the estimated times for completing the remaining phases (e.g., the injection phase and the retraction phase) divided by the number of unlit LEDs in the progress loop. After the injection phase is completed, the LEDs can be activated at a rate equal to the estimated time for completing the retraction phase divided by the number of unlit LEDs.

[0162] In some implementations, a subset of LEDs can be used to indicate the progress of the injection phase. For example, in an implementation with eight LEDs positioned on the housing of the autoinjector 2, the first LED can be illuminated to indicate needle insertion. Then, the second through seventh LEDs can be illuminated sequentially to indicate the progress of the injection phase. Finally, the eighth LED can be illuminated to indicate needle retraction. While exemplary configurations of LEDs and corresponding logic have been described, it should be understood that the number of LEDs used for each phase of the injection process can be varied as needed.

[0163] The vision module 1420 may also include a display screen, touchscreen, or other suitable device to provide one-way or two-way communication with the user. The user can see the vision module 1420 from the outside of the housing 3 through a window on the housing 3. The haptic module 1422 may include, for example, a haptic motor configured to generate vibrations that the user can feel. The vibrations may signal the start and end of the injection and / or may help provide additional information to the user.

[0164] The controller 1408 can be coupled to a wireless communication module and an antenna. The wireless communication module can be configured to transmit data from the controller 1408 to devices such as mobile devices, computers, and mobile phones. The wireless communication module can be configured to transmit information via one or more wireless modes (e.g., Bluetooth, Bluetooth Low Energy (BLE), Near Field Communication (NFC), infrared, cellular networks, and wireless networks). The antenna can be any suitable device configured to assist the wireless communication module in data transmission and / or amplification. Therefore, the controller 1408 can be configured to transmit diagnostic information about the user and / or the autoinjector 2, information related to injection completion, and / or information related to the malfunction status of the autoinjector 2 to the user's device or the cloud. Signals indicating needle insertion and / or early device removal can also be transmitted via the wireless communication module. The controller 1408 can also be configured to transmit temperature information of the autoinjector 2. For example, when the autoinjector 2 is removed from a refrigerator, the user can monitor, for example, the temperature of the autoinjector 2 via a mobile device and / or an application. The controller 1408 can also receive activation and / or delay commands via the wireless communication module. For example, the controller 1408 can also receive operation adjustment commands, such as commands related to adjusting the preferred operating speed. In some implementations, the controller 1408 may receive a command to pause the injection.

[0165] In some embodiments, the controller 1408 can communicate with a mobile application on a user's mobile device via a wireless communication module. The mobile application can be configured to facilitate use of the autoinjector 2 and improve the user experience. In some embodiments, the mobile application can be used to automatically check the expiration date of the medicine contained in the autoinjector 2. Such functionality frees the user from manually checking the expiration date and can improve user safety. Based on the expiration date, the mobile device can be configured to remind the user and / or disable the use of the autoinjector 2. In some embodiments, the mobile application can be used to remind the user of a product recall and / or disable the device in the event of a product recall. For example, the mobile application can access a database via the Internet to determine whether a specific device, a large number of devices, medicines, and / or a large number of medicines have been recalled. In some embodiments, the mobile application can be configured to verify whether the autoinjector 2 and / or the medicine are genuine and not counterfeit. For example, the mobile application can achieve this by cross-referencing the product serial number or digital signature with a database of certified products. In some embodiments, one or more parts of the autoinjector 2 can be disposable, and the mobile application can be configured to verify the authenticity of such one or more parts before use.

[0166] In some embodiments, the mobile application can be used to facilitate the injection sequence. For example, the mobile application can be synchronized with the events of the injection sequence and provide the user with simultaneous instructions on when to perform which tasks (e.g., pressing switch 1409, holding the autoinjector 2 on the skin, removing the autoinjector). In some embodiments, the instructions can be spoken. In some embodiments, the instructions can be provided visually via a display on the mobile device. In some embodiments, the mobile application can be configured to provide detailed indications of the progress of the injection sequence. For example, the mobile application can provide textual, visual, and / or auditory progress indications with finer detail than those shown by LEDs, such as those previously described herein.

[0167] In some implementations, the mobile application can be configured to record and store the date and / or time of the injection. Based on the date and / or time of the injection and the user's prescription information, the mobile application can be configured to automatically create reminders for subsequent injections. In some implementations, upon completion of the injection, the mobile application can be configured to provide the user with a notification containing positive feedback regarding adherence to the prescription protocol. In some implementations, the mobile application can offer points and / or rewards for continued adherence.

[0168] In some implementations, the mobile application can be configured to authenticate the user of the autoinjector 2 before use. For example, the mobile application, combined with the user's mobile device, can use biometrics, two-factor authentication, or any other suitable authentication protocol to verify the user's identity before injection. Upon authenticating the user, the mobile application can activate or otherwise unlock the autoinjector. This user authentication can prevent misuse and / or waste of expensive medication by people other than the intended user.

[0169] In some implementations, the mobile application can be configured to detect the operating conditions of the autoinjector 2. For example, the mobile application can be configured to detect the device's battery level and, upon a low battery indication, to provide a notification to the user indicating that the device needs to be charged. In some implementations, the mobile application can be configured to detect mechanical and / or electrical faults in the autoinjector 2 and communicate such information to the user.

[0170] Figure 18 An exemplary method 2000 according to this disclosure is illustrated. Method 2000 may begin at step 2002, where a user positions an autoinjector 2 on her body such that the tissue-engaging surface 4 contacts the skin surface. The user may position the autoinjector 2 on her skin after removing the locking member 1610, as previously described herein, thereby allowing the touch sensor 1410 to detect proximity to the skin. The autoinjector 2 may be mounted in any suitable location, such as the thigh, abdomen, shoulder, forearm, upper arm, leg, hip, or another suitable location. The autoinjector 2 may be secured to the skin by an adhesive patch 12. The securing of the autoinjector 2 at step 2002 may cause an activation switch 1409 extending outward from the tissue-engaging surface 4 to be pressed and disconnect the circuit. The disconnection of the circuit may cause a signal indicating that the activation switch 1409 has been pressed to the controller 1408. Alternatively, any other suitable mechanism may be used to energize or otherwise activate the autoinjector 2 before or after step 2002. When the activation switch 1409 is pressed, the auto-injector 2 may emit an audio tone and / or illuminate one or more LEDs (e.g., one or more LEDs of a first color (e.g., blue)) to indicate that the activation switch 1409 has been pressed.

[0171] Once the autoinjector 2 is activated at step 2002, method 2000 can proceed to step 2004, where controller 1408 can determine whether tissue-engaging surface 4 is positioned on the skin surface. At step 2004, controller 1408 can receive a measurement from touch sensor 1410 indicating whether the autoinjector 2 is positioned on the skin or another surface. If controller 1408 determines that touch sensor 1410 is in contact with the skin, for example, when the capacitance value received from touch sensor 1410 is within a predetermined range, method 2000 can proceed to step 2008. If controller 1408 determines that touch sensor is not in contact with the skin, for example, if the capacitance measurement received from touch sensor 1410 indicates that autoinjector 2 is in contact with a non-skin surface such as wood or metal, method 2000 can proceed to step 2006. At step 2006, autoinjector 2 may be placed in an error state. In the error state, an LED (e.g., a red LED) can be activated to indicate to the user that an error has occurred, or a message can be displayed on the screen. In some instances, the autoinjector 2 may need to be manually reset before the injection can be completed. In other instances, the autoinjector 2 may loop back to step 2004, where the controller 1408 continuously attempts to determine whether the touch sensor 1410 is in contact with the skin. Method 2000 may also require the touch sensor 1410 to be in contact with the skin throughout the injection. Thus, if at any point during the injection, the controller 1408 determines that the touch sensor 1410 is no longer in contact with the skin, the controller 1408 may stop the injection (e.g., by stopping further movement of the translation mechanism 1366), may generate an error signal or message, and may retract the needle 306 if it has been extended. By stopping the injection and retracting the needle 306, the risk of dispensing medication outside the body (i.e., wet injection) and / or needlestick injury can be mitigated. After determining step 2004, the autoinjector 2 may emit an audio tone and / or illuminate one or more LEDs to indicate that the autoinjector 2 is positioned on the skin surface. In one instance, one or more additional LEDs of a first color may be illuminated at this stage to indicate further progress of the injection.

[0172] At step 2008, controller 1408 may send a signal to activate translation mechanism 1366. Once activated, translation mechanism 1366 may move toward the second end 1306 of cartridge 1302 (see...). Figure 13 and Figure 14 This causes the cartridge 1302 itself to move in the same direction. This allows the needle 308 to move in the opposite direction to its entry into the cartridge 1302 as described above. The movement of the actuator 1398 and the needle 308 causes the carrier 202 to move in the same direction, which illustrates the effect of... Figures 5 to 11The mechanism described herein ultimately deploys needle 306 into the user's body in a chain of events. The translation mechanism 1366 will continue moving toward the second end 1306 until the desired amount of medication contained in cartridge 1302 has been dispensed into the user's body. When the translation mechanism is activated, the autoinjector 2 may emit an audio tone and / or illuminate one or more LEDs to indicate that injection is in progress. For example, additional LEDs of a first color may be illuminated during injection to provide the user with a visual indication of progress.

[0173] Method 2000 can proceed to step 2010, where controller 1408 can determine whether the injection is complete. This determination can be based on the interruption of beam 1430 by piston 1316 (as referenced). Figure 4A , Figure 13 and Figure 14 (As described above). That is, when the beam 1430 is disrupted (not received by the detector 1416), the controller 1408 can determine that the injection is complete. Once the controller 1408 determines that the injection is complete, the controller 1408 can send a signal to the translation mechanism 1366 to reverse the rotation direction of the lead screw, which causes the ramp 1500 to push against the ramp 243 of the stop 240, enabling the needle 306 to retract, as described above. Figure 11 As discussed. In one example, controller 1408 may establish a delay after receiving an indication that beam 1430 has been interrupted. The delay may be, for example, 0.1 seconds to 60 seconds.

[0174] An additional end detection mechanism can be used in place of or in combination with the aforementioned interruption-type sensor. For example, the current of the motor of the translation mechanism 1366 can be used to determine whether the injection has been completed. That is, when the piston 1316 reaches the second end 1306 of the cartridge 1302, the current on the motor will increase (e.g., because the piston 1316 engages the end of the cartridge 1302), thereby signaling the discharge of all or substantially all of the contents of the cartridge 1302. An exemplary combination may include the use of a light beam 1430, wherein an interruption of the light beam 1430 indicates, for example, that 90% to 98% of the injection has been completed. The current of the motor of the translation mechanism 1366 can then be analyzed to determine whether the remaining 2% to 10% of the injection has been completed. In another example, instead of using a light switch, the controller 1408 can use a delay from the start of the translation mechanism 1366 to determine when to reverse the translation mechanism 1366. In one example, this delay may be, for example, from about 1 second to about 120 seconds, but other suitable times may also be expected. In any case, the delay from start-up can be long enough to allow cartridge 1302 to be emptied. In yet another example, beam 1430 can be used in conjunction with an encoder. The encoder can be configured to detect the positioning of piston 1316. If the encoder is used only to detect the positioning of piston 1316, drivetrain issues may hinder accurate detection. For example, piston 1316 may rotate when driven by a leadscrew. This rotation may cause the actual positioning of piston 1316 to be uncertain. However, when used in conjunction with beam 1430, controller 1408 can be configured to recalibrate the encoder in response to an interruption of beam 1430. This recalibration allows controller 1408 to update the actual positioning of the encoder and restore accurate detection of the positioning of piston 1316 using the encoder.

[0175] Upon confirmation of injection completion, the autoinjector 2 may emit an audio tone and / or illuminate one or more LEDs to indicate that the injection is complete. In some instances, one or more LEDs of a second color (e.g., green) different from the first color may be illuminated to signal to the user that the injection is complete. In some instances, all LEDs of the device may be illuminated in the second color, and other indicators may also be used. For example, all LEDs may be illuminated in the second color and may flash intermittently at the end of the injection.

[0176] In some instances, the timing of the injection process, measured from the initial activation of the activation switch 1409 to the retraction of the needle 306 from the user after drug delivery, can be approximately 20 seconds to approximately 90 seconds, or approximately 25 seconds to approximately 60 seconds, approximately 30 seconds to approximately 45 seconds, or less than or equal to approximately 120 seconds, or less than or equal to approximately 90 seconds, or less than or equal to approximately 60 seconds, or less than or equal to approximately 45 seconds, or less than or equal to approximately 30 seconds. This timing represents a significant improvement over existing devices, for which the injection timing can be much longer, and in some cases up to approximately 9 minutes or even longer.

[0177] Method 2000 may also include additional steps. For example, method 2000 may include determining whether the medication in cartridge 1302 is too cold to be delivered to the user, whether power supply 1406 has sufficient power to complete the injection, whether needle 306 has been prematurely deployed and / or retracted, whether the current of the motor of translation mechanism 1366 is within the appropriate range, and whether the injection process has been prolonged beyond the maximum acceptable process time. When controller 1408 senses any of the above errors, it can communicate such errors to the user and can terminate the ongoing injection by, for example, stopping or reversing translation mechanism 1366 and retracting needle 306 from the user. The autoinjector 2 may emit an audio tone and / or illuminate one or more LEDs indicating any of the aforementioned additional steps. For example, one or more LEDs of a third color (e.g., red) different from the first and second colors may be illuminated.

[0178] Figure 20 An exemplary method 2020 is shown for controlling the torque of the motor of the translation mechanism 1366 and detecting when the motor stalls. At step 2022, the controller 1408 may initiate an injection sequence. As previously described herein, the injection sequence may be initiated when the activation switch 1409 is pressed against the user's skin and / or when the user's skin is detected by the touch sensor 1410. During the injection sequence, voltage may be applied to the motor of the translation mechanism 1366 to drive the motor.

[0179] At step 2024, as the injection sequence proceeds, controller 1408 can maintain the motor of translation mechanism 1366 at a constant speed. The constant speed can be, for example, a rotational speed measured in revolutions per minute (RPM). Controller 1408 can maintain the motor at a constant speed by changing the voltage applied to the motor. For example, when a higher load is applied to the motor due to blockage, increased fluid pressure, increased component friction, or any other reason, controller 1408 can compensate for the increased load by increasing the voltage applied to the motor. Conversely, when the load applied to the motor decreases, controller 1408 can compensate for the decreased load by decreasing the voltage applied to the motor. Maintaining the motor at a constant speed can reduce the likelihood of the user experiencing injection site pain. For example, maintaining the motor at a constant speed can prevent the injection from becoming too large, thereby reducing the risk of pain.

[0180] During the injection sequence, controller 1408 can monitor the current supplied to the motor. The motor current can indicate the torque generated by the motor. For example, a higher motor current can indicate a larger torque generated by the motor. At step 2026, controller 1408 can determine whether the motor current exceeds a first current threshold. The first current threshold can be determined and / or set based on the maximum torque that can be safely generated by the motor. For example, the maximum torque can be reached when the injection sequence is somehow blocked. If controller 1408 determines that the motor current does not exceed the first current threshold, method 2020 can return to step 2024, and controller 1408 can continue to maintain the motor at a constant speed. Alternatively, if controller 1408 determines that the motor current exceeds the first current threshold, method 2020 can proceed to step 2028.

[0181] At step 2028, controller 1408 may reduce the motor voltage to maintain the motor current below a second current threshold. In some embodiments, the second current threshold may be greater than the first current threshold and may be more closely related to the maximum torque that can be safely generated by the motor. In some embodiments, the second current threshold may be less than or equal to the first current threshold. In the event of an injection sequence disruption, the motor speed may slow down and the motor impedance may decrease. As the motor impedance decreases, a lower voltage may be required to maintain the motor current below the second current threshold. Controller 1408 may monitor the average motor voltage applied to the motor. The average motor voltage may be, for example, a time-averaged value.

[0182] Steps 2020 of Method 2020 from 2024 to 2028 can usually be achieved through... Figure 20B The graph depicted in the figure illustrates the relationship between the voltage applied to the motor of the translation mechanism 1366 and the current consumed by the motor. This graph can be characterized by the following equation:

[0183] V = iR + V emf

[0184] In the above formula, V is the voltage applied to the motor, i is the current consumed by the motor, R is the resistance of the motor coil, and V emf It is the back electromotive force acting on the applied voltage at a given velocity. For example... Figure 20B As shown, the curve can include a constant speed region, where the motor can maintain a constant speed (step 2024). In the constant speed region, V emf It can remain approximately constant, and the curve can be approximately linear.

[0185] like Figure 20B As shown, when the load (i.e., torque) on the motor used for the translation mechanism 1366 increases, the current consumed by the motor can increase. When the current approaches the maximum current, the voltage applied to the motor can be reduced, thereby keeping the current below the maximum current (steps 2026 and 2028). Proportional-integral (PI) regulation can be used to keep the current consumed by the motor below the maximum current. As shown, there may be a minimum voltage for the motor; below this minimum voltage, the motor may stall.

[0186] Steps 2030 to 2038 of method 2020 may correspond to a control sequence for preventing motor stall. Figure 20A A graph was plotted that can represent the voltage applied to the motor and the current consumed by the motor over time, as well as the curves according to steps 2030 to 2038.

[0187] At step 2030, controller 1408 may determine whether the average motor voltage has decreased below a first threshold voltage. An average motor voltage decreasing below the first threshold voltage may indicate that the injection sequence is blocked. If controller 1408 determines that the average motor voltage has not decreased below the first threshold voltage, method 2020 may return to step 2028, where controller 1408 may continue to maintain the motor current below a second current threshold. Figure 20A The diagram illustrates five intervals in which the controller 1408 can maintain the motor current at a constant value (e.g., below a second current threshold): between approximately 35 seconds and approximately 37 seconds, between approximately 39 seconds and approximately 41.5 seconds, between approximately 43.5 seconds and approximately 46 seconds, between approximately 48 seconds and approximately 50.5 seconds, and between approximately 52.5 seconds and approximately 55 seconds. Figure 20A As shown, the voltage applied to the motor during each interval can be reduced (albeit with some fluctuations) to keep the motor current below the second current threshold. Although Figure 20A The voltage during each interval is shown as decreasing, but in order to keep the motor current below the second current threshold, the voltage does not necessarily need to decrease, and in some cases it can remain stable.

[0188] On the other hand, if the controller 1408 determines that the average motor voltage has decreased below a first threshold voltage, the controller 1408 may pause the injection sequence for a first time interval. When the injection sequence is paused, the controller 1408 may stop applying voltage to the motor. In some embodiments, the first time interval may be, for example, 2 seconds. Figure 20A Four such pauses are shown: between approximately 37 and approximately 39 seconds, between approximately 41.5 and approximately 43.5 seconds, between approximately 46 and approximately 48 seconds, and between approximately 50.5 and approximately 52.5 seconds.

[0189] The first time interval can be long enough to allow the fluid pressure within the autoinjector 2 to dissipate. The first time interval can also be short enough that the autoinjector 2 can be removed from the user's skin without prompting the user (e.g., the first time interval is set to be less than the typical reaction time for a user to mistakenly recognize the end of the injection). The first time interval can also be indicated by illuminating one or more LEDs or another light on the progress ring within the autoinjector 2, and this should be visible to the user. The LEDs can be illuminated, for example, in a specific pattern or according to a specific color scheme, to indicate a pause rather than a stop in the injection sequence during the first time interval.

[0190] After pausing the injection sequence, controller 1408 can resume the injection sequence at step 2034. To resume the injection sequence, controller 1408 can restore the voltage supply to the motor of translation mechanism 1366. At step 2036, controller 1408 can determine whether the average motor voltage has decreased below a first threshold voltage within a second time interval. The second time interval may be shorter than the first time interval and may be set and / or determined to indicate confirmation that the injection sequence has been blocked. The second time interval may be, for example, about 0.9 seconds. If the motor voltage has not decreased below the first threshold voltage within the second time interval, method 2020 can return to step 2030. Alternatively, if controller 1408 determines that the motor voltage has decreased below the first threshold voltage within the second time interval, method 2020 can proceed to step 2038, where controller 1408 can abort the injection sequence.

[0191] In some implementations, the controller 1408 may continue executing step 2026 while performing steps 2028 to 2036. For example, while performing steps 2028 to 2036, the controller 1408 may continue to determine whether the motor current exceeds a first current threshold. If the motor current continues to exceed the first current threshold, method 2020 may proceed through steps 2028 to 2036 as previously described herein. Alternatively, if the motor current drops below the first current threshold, method 2020 may return to step 2024, and the controller 1408 may maintain the motor at a constant speed. In other words, if the high load on the motor due to blockage, high fluid pressure, etc., dissipates during the execution of steps 2028 to 2036, the controller 1408 may simply resume maintaining a constant motor speed instead of unnecessarily proceeding through any remaining steps.

[0192] Therefore, method 2020 allows controller 1408 to effectively distinguish between situations where the needle may be partially blocked or high friction may act on the injection sequence, and situations where the injection sequence is subject to insurmountable blockage. In the former case, autoinjector 2 may have the capability to complete the injection sequence, and the injection sequence may not be prematurely terminated. In the latter case, autoinjector 2 may not have the capability to complete the injection sequence, and the injection sequence may be appropriately terminated. In such cases, autoinjector 2 may emit an audio tone and / or illuminate one or more LEDs to indicate that the injection was terminated before completion. Method 2020 may also appropriately terminate the injection sequence when piston 1316 is fully extended, indicating that cartridge 1302 is empty. Method 2020 may also allow autoinjector 2 to be used in emergency situations, for example, if the user performs the injection without first preheating autoinjector 2 to reduce drug viscosity. Method 2020 may also allow the injection of viscous drugs at a slower rate than that otherwise permitted by motor and gear reduction ratios.

[0193] Figure 21 An exemplary method 2100 for detecting the end of a drug dose using a transmitter 1414 and a detector 1416 is shown. Method 2100 can be used, for example, to detect when a full dose of drug has been administered to a user and the corresponding injection sequence has ended.

[0194] At step 2102, controller 1408 may initiate the injection sequence. As previously described herein, the injection sequence may be initiated when the user's skin is pressed against the activation switch 1409 and / or when the user's skin is detected by touch sensor 1410. At step 2104, controller 1408 may periodically cycle the transmitter 1414 on and off. The transmitter 1414 may cycle on and off rapidly in a square wave mode, such that the transmitter 1414 cycles on and off several times per second. Cyclicly cycling the transmitter 1414 on and off allows detector 1416 to be exposed to light generated by transmitter 1414 in combination with ambient light, and also to ambient light alone.

[0195] At step 2106, the controller 1408 may receive from the detector 1416 a first signal corresponding to the time when the transmitter 1414 is turned off. The first signal may correspond to and / or indicate the ambient light detected by the detector 1416. At step 2108, the controller 1408 may receive from the detector 1416 a second signal corresponding to the time when the transmitter 1414 is turned on. The second signal may correspond to and / or indicate the light emitted by the transmitter 1414 in combination with the ambient light detected by the detector 1416.

[0196] At step 2110, controller 1408 can calculate the difference between a first light value represented by a first signal and a second light value represented by a second signal. This difference can indicate how much of the light detected by detector 1416 is attributable to light emitted by emitter 1414 rather than ambient light. At step 2112, controller 1408 can determine whether the difference is less than a threshold. If controller 1408 determines that the difference is not less than the threshold, method 2100 can return to step 2106. Alternatively, if controller 1408 determines that the difference is less than the threshold, controller 1408 can terminate the injection sequence at step 2114.

[0197] Therefore, method 2100 can be used to reduce the influence of ambient light when detecting the end of a drug dose. Specifically, method 2100 can address situations where light from transmitter 1414 is blocked and cannot reach detector 1416 indicating the end of the dose, while ambient light can reach detector 1416 and produce a false negative reading indicating that the end of the dose has not yet been reached.

[0198] Figure 22 Another exemplary method 2200 for detecting the end of a drug dose is shown, using a transmitter 1414 and a detector 1416. Method 2200 can be used, for example, to detect when a full dose of drug has been administered to a user and the corresponding injection sequence has ended.

[0199] At step 2202, controller 1408 may initiate the injection sequence. As previously described herein, the injection sequence may be initiated when the user's skin is pressed against the activation switch 1409 and / or when the user's skin is detected by touch sensor 1410. At step 2204, controller 1408 may activate transmitter 1414 or otherwise cause transmitter 1414 to emit light.

[0200] At step 2206, controller 1408 may allow the injection sequence to continue for a first time period. The first time period may be a predetermined time period corresponding to a duration during which it is impossible or unlikely to administer the full dose. For example, the first time period may be between approximately 20% and 50% of the total injection time. During the first time period, controller 1408 may not interrupt the injection sequence in response to a signal received from detector 1416 (but may still interrupt it due to factors such as reference...). Figure 20 The discussed blockage or stall that interrupts the injection sequence.

[0201] At step 2208, after the first time period ends, controller 1408 can determine whether the amount of light received by detector 1416 is less than a first threshold light value. Controller 1408 can make this determination based on a signal received from detector 1416 indicating the amount of light received by detector 1416. The first threshold light value may correspond to the amount of light received by detector 1416 at the end of the dose. If controller 1408 determines that the amount of light received by detector 1416 is not less than the first threshold light value, controller 1408 can continue the injection sequence, and method 2200 can otherwise remain at step 2208. Alternatively, if controller 1408 determines that the amount of light received by detector 1416 is less than the first threshold light value, the method can proceed to step 2210.

[0202] At step 2210, controller 1408 can determine whether the amount of light received by detector 1416 is greater than or equal to a first threshold light value. If controller 1408 determines that the amount of light received by detector 1416 has risen to or above the first threshold light value, controller 1408 can continue the injection sequence, and method 2200 can return to step 2208. On the other hand, if controller 1408 determines that the amount of light received by detector 1416 has remained below the first threshold light value, the method can proceed to step 2212. Step 2210 can effectively enable controller 1408 to "clear" injection sequences with anomalous interruptions in the light received by the detector, which may be caused by air bubbles in cartridge 1302 that block the path of light between transmitter 1414 and detector 1416, for example, provided that the light amount subsequently reaches or exceeds the first threshold light value.

[0203] At step 2212, controller 1408 may determine whether the motor current exceeds a first threshold current value. The first threshold current value may be determined and / or set based on the current indicating the end of the injection sequence. For example, the first current threshold may be set based on the current indicating that the piston 1316 has reached the second end 1306 of the cartridge 1302. If controller 1408 determines that the motor current does not exceed the first threshold current value, controller 1408 may continue the injection sequence, and method 2200 may return to step 2208. Alternatively, if controller 1408 determines that the motor current exceeds the first threshold current value, method 2200 may proceed to step 2214, at which step controller 1408 may end the injection sequence.

[0204] Therefore, method 2200 allows for accurate identification of the end of the injection sequence by recognizing the moment when both the light received by detector 1416 and the motor current indicate the end of the dose. By sequentially performing steps 2208, 2210, and 2212, erroneous identification of the end of the dose due to abnormal interruption of light or a single abnormally high current event can be mitigated. Method 2200 can specifically reduce the influence of air bubbles within cartridge 1302 on the detection of the end of the drug dose.

[0205] Figure 23 An exemplary method 2300 for operating the activation switch 1409 of the autoinjector 2 according to the present disclosure is shown. Specifically, Figure 23 An exemplary sequence of functions for positioning the activation switch 1409 and the corresponding functions for the autoinjector 2 is depicted.

[0206] Initially, at step 2302, the autoinjector 2 can be placed within the packaging such that the plunger 1450 is pressed and the autoinjector 2 is in a low-power sleep mode. In some embodiments, the autoinjector 2 can be programmed to be in a wake-up or activated state during manufacturing. In some embodiments, the autoinjector 2 can be configured to switch to a low-power sleep mode if the plunger 1450 is pressed for a predetermined period of time after programming (such as when the autoinjector is placed in the packaging). The predetermined period of time can be any suitable period of time, such as 60 minutes, 30 minutes, 15 minutes, 10 minutes, 5 minutes, 2 minutes, or any other suitable period of time. The autoinjector 2 can be sealed in the packaging such that the packaging indicates that the autoinjector 2 has not been previously used. The packaging can be made of any suitable material, including paper, cardboard, plastic, cellophane, etc. The packaging can press against the plunger 1450 such that the plunger 1450 is flush or nearly flush with the housing 3 of the autoinjector 2 and prevents the plunger 1450 from protruding outward from the autoinjector 2. When the plunger 1450 is in the pressed state, the circuit associated with the activation switch 1409 can be disconnected, thereby maintaining the autoinjector 2 in a low-power sleep mode.

[0207] At step 2304, the autoinjector 2 can be removed from the packaging, so that the plunger 1450 is no longer pressed down by the packaging and can extend outward from the autoinjector 2. When the plunger 1450 changes from the pressed state to the free or extended state, the plunger flange 1454 can contact or otherwise press the plunger switch 1448, thereby completing the circuit associated with the activation switch 1409.

[0208] At step 2306, in response to the completion of the circuitry associated with activation switch 1409, the auto-injector transitions from a low-power sleep mode to an active mode. In active mode, the auto-injector 2 can calibrate touch sensor 1410. The auto-injector 2 can calibrate touch sensor 1410 by detecting a value or measurement of touch sensor 1410 in ambient air (i.e., not in contact with the user's skin). This calibration can be performed during a predetermined period after the auto-injector 2 has been removed from its packaging (in some cases, immediately after removal), such that the calibration occurs before the user exposes their skin to touch sensor 1410. In active mode, the auto-injector 2 can also detect whether the transmitter 1414 and / or detector 1416 are functioning properly, whether the needle is correctly positioned, whether the motor of translation mechanism 1366 is responsive and / or operable, and / or perform any other suitable status tests. The auto-injector 2 can detect needle positioning, for example, using a switch or detector configured to report needle positioning to controller 1408. In active mode, the autoinjector 2 can also illuminate one or more backlights to allow the user to inspect the vial and / or the medication contained therein through the transparent window 50. In active mode, the autoinjector 2 can also display any other indications that it is ready for use.

[0209] At step 2308, the auto-injector 2 can be placed against the user's skin, causing the plunger 1450 to be pressed into the auto-injector 2. When the plunger 1450 is pressed, the circuit associated with the activation switch 1409 can switch to the off state. (Refer to the above text.) Figure 18 As described in method 2000, the autoinjector 2 may also use a touch sensor 1410 to detect contact with the skin. In response to the pressing of the plunger 1450 and the detection of contact with the skin, the autoinjector 2 may initiate an injection sequence at step 2310. The injection sequence may be a sequence that causes the user to inject a drug, as previously described herein.

[0210] At step 2312, the autoinjector 2 can be removed from the user's skin, and the plunger 1450 can be extended outward again from the autoinjector 2. As the plunger 1450 extends outward, the circuit associated with the activation switch 1409 can transition from an open state to a closed state. In response, the autoinjector 2 can end the injection sequence at step 2310 and initiate the retraction of the patient needle, for example, by reversing the motor. If the injection sequence has been completed, or if the autoinjector has been prematurely or accidentally removed from the skin, the autoinjector 2 can initiate needle retraction to prevent wet injection. Alternatively, in some embodiments, the controller 1408 can determine whether a value received from the touch sensor 1410 indicates that the autoinjector 2 remains in contact with the user's skin. If a value received by the controller 1408 indicates that the autoinjector 2 remains in contact with the user's skin, the autoinjector 2 can pause the injection sequence, thereby preventing wet injection. If the plunger 1450 is pressed again, thereby disconnecting the circuit associated with the activation switch 1409, the autoinjector 2 can resume the injection sequence.

[0211] According to the aforementioned method 2300, the activation switch 1409 can be used to keep the autoinjector 2 in a low-power sleep mode while in packaging, switch the autoinjector 2 to an active mode when removed from packaging, indicate when to place the autoinjector 2 on the user's skin for an injection sequence, and indicate when to remove the autoinjector 2 from the user's skin at the end of the injection sequence. Furthermore, the signal from the activation switch 1409 can be cross-checked with the signal from the touch sensor 1410 to more accurately determine whether the autoinjector 2 has been removed from the user's skin, or, for example, whether an accidental or slight movement of the autoinjector has occurred.

[0212] It should be understood that in some embodiments, one or more steps of the various methods described herein may be combined. Furthermore, in some embodiments, fewer than all the steps of the methods described herein may be performed and / or additional steps not described herein may be performed. Moreover, the steps described herein do not necessarily need to be performed in the exact order presented.

[0213] Medicaments and drugs

[0214] The embodiments described herein can be used in conjunction with the administration of various pharmaceutical products, drugs, and / or pharmaceutical preparations to a patient. Exemplary pharmaceutical products, drugs, and / or pharmaceutical preparations that can be used with the embodiments described herein are described in U.S. Patent Nos. 8,945,559B2, 9,987,500B2, and 11,603,407B2, the entire contents of which are incorporated herein by reference. Pharmaceutical products, drugs, and / or pharmaceutical preparations that can be used with the embodiments of this disclosure are further described in detail below.

[0215] First, exemplary pharmaceutical products, drugs, and / or pharmaceutical formulations conforming to U.S. Patent No. 8,945,559B2 are described. As used herein, the term "pharmaceutical formulation" means a combination of at least one active ingredient (e.g., a small molecule, macromolecule, compound, etc., capable of exerting biological effects in a human or non-human animal) and at least one inactive ingredient suitable for therapeutic administration to a human or non-human animal when combined with the active ingredient or one or more additional inactive ingredients. As used herein, unless otherwise specifically stated, the term "formulation" means "pharmaceutical formulation".

[0216] This disclosure provides pharmaceutical formulations comprising at least one therapeutic polypeptide. According to certain embodiments of this disclosure, the therapeutic polypeptide is an antibody or antigen-binding fragment thereof that specifically binds to human interleukin-4 receptor α (hIL-4Rα). More specifically, this disclosure includes pharmaceutical formulations comprising: (i) a human antibody that specifically binds to hIL-4Rα; (ii) an acetate / histidine buffer system; (iii) an organic cosolvent that is a nonionic surfactant; (iv) a heat stabilizer for carbohydrates; and (v) a viscosity reducer. Specific exemplary components and formulations included in this disclosure are described in detail below.

[0217] The pharmaceutical formulations disclosed herein may comprise a human antibody or antigen-binding fragment thereof that specifically binds to hIL-4Rα. As used herein, the term "hIL-4Rα" refers to a human cytokine receptor that specifically binds to interleukin-4 (IL-4). In some embodiments, the antibody contained in the pharmaceutical formulations of this disclosure specifically binds to the extracellular domain of hIL-4Rα. An exemplary amino acid sequence of human IL-4 receptor α (hIL-4Rα) is described in SEQ ID NO:25. Antibodies against hIL-4Rα are described in U.S. Patent Nos. 7,605,237 and 7,608,693. The extracellular domain of hIL-4Rα is represented by the amino acid sequence of SEQ ID NO:26.

[0218] As used herein, the term "antibody" generally refers to an immunoglobulin molecule comprising four polypeptide chains (two heavy (H) chains and two light (L) chains) linked together by disulfide bonds, as well as its polymers (e.g., IgM); however, immunoglobulin molecules consisting only of heavy chains (i.e., lacking light chains) are also included within the definition of the term "antibody." Each heavy chain contains a heavy chain variable region (abbreviated herein as HCVR or V). H The heavy chain constant region contains three domains: CH1, CH2, and CH3. Each light chain contains a light chain variable region (abbreviated as LCVR or V in this paper). LThe light chain constant region contains a structural domain (CL1). H District and V L The region can be further subdivided into highly variable regions (called complementary determinant regions (CDRs)) and more conservative regions scattered between them (called frame regions (FRs)). Each V H and V L It consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0219] Unless otherwise specified, as used herein, the term “antibody” should be understood to encompass the complete antibody molecule and its antigen-binding fragment. As used herein, the term “antigen-binding portion” or “antigen-binding fragment” (or simply “antibody portion” or “antibody fragment”) of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to hIL-4Rα or its epitopes.

[0220] As used herein, “isolated antibody” is intended to refer to an antibody that is substantially free of other antibodies that have different antigen specificities (e.g., an isolated antibody that specifically binds to hIL-4Rα is substantially free of antibodies that specifically bind to antigens other than hIL-4Rα).

[0221] The term "specific binding" refers to the formation of a relatively stable complex between an antibody or its antigen-binding fragment and the antigen under physiological conditions. Specific binding can be achieved by a dissociation constant of at least approximately 1 × 10⁻⁶. -6 M or larger is used for characterization. Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, etc. However, isolates that specifically bind hIL-4Rα may cross-react with other antigens, such as IL-4R molecules (orthologs) from other species. In the context of this disclosure, multispecific (e.g., bispecific) antibodies that bind hIL-4Rα and one or more additional antigens are considered to "specifically bind" hIL-4Rα. Furthermore, isolates may be substantially free of other cellular material or chemicals.

[0222] Exemplary anti-hIL-4Rα antibodies that may be included in the pharmaceutical formulations disclosed herein are described in U.S. Patent Nos. 7,605,237 and 7,608,693, the disclosure of which is incorporated herein by reference in its entirety.

[0223] According to certain embodiments of this disclosure, the anti-hIL-4Rα antibody is human IgG1 comprising the heavy chain variable region of the IGHV3-9 subtype and the light chain variable region of the IGKV2-28 subtype (see Barbie and Lefranc, The Human Immunoglobulin Kappa Variable (IGKV) Genes and Joining (IGKJ) Segments, Exp. Clin. Immunogenet. 1998; 15:171-183; and Scaviner, D. et al., Protein Displays of the Human Immunoglobulin Heavy, Kappa and Lambda Variable and Joining Regions, Exp. Clin. Immunogenet. 1999; 16:234-240).

[0224] In some embodiments, anti-hIL-4Rα comprises at least one amino acid substitution that results in a charge change at the exposed surface of the antibody relative to the germline IGHV3-9 sequence or the germline IGKV2-28 sequence. The germline IGHV3-9 and IGKV2-28 sequences and amino acid position assignments provided herein conform to the International Immunogenetics (IMGT) Information System, such as Lefranc, M.-P. et al. the international ImMunoGeneTicsInformation As described in Nucl. Acids Res, 37, D1006-D1012 (2009). In some embodiments, the exposed surface comprises a complementarity-determining region (CDR). In some embodiments, one or more amino acid substitutions are selected from the group consisting of: (a) basic amino acid substitutions for neutral amino acids within CDR2 (e.g., position 58) of IGHV3-9; (b) neutral amino acid substitutions for acidic amino acids within CDR3 (e.g., position 107) of IGHV3-9; and (c) neutral amino acid substitutions for basic amino acids within CDR1 (e.g., position 33) of IGKV2-28. The unique arrangement of the antibody's charge distribution, particularly at the environmental interface (e.g., in the CDR), is expected to create unpredictable conditions for the antibody's stability in solution.

[0225] In some embodiments, the anti-hIL-4Rα antibody comprises at least one amino acid substitution that produces a torsional strain change relative to the germline IGHV3-9 sequence or the germline IGKV2-28 sequence within the frame region of the antibody's variable region. In some embodiments, one or more amino acid substitutions are selected from the group consisting of: (a) proline substitution for a non-proline amino acid in frame region 3 (FR3) (e.g., position 96) of IGHV3-9; and (b) non-proline substitution for a proline in frame region 2 (FR2) (e.g., position 46) of IGKV2-28. Changes in peptide chain rotation capability, particularly within the frame region, affect the interface between the CDR and the solvent, and are expected to create unpredictable conditions for the antibody's stability in solution.

[0226] According to certain embodiments of this disclosure, the anti-hIL-4Rα antibody or its antigen-binding fragment comprises the heavy chain complementarity-determining region (HCDR) 1 of SEQ ID NO:2, HCDR 2 of SEQ ID NO:3, and HCDR 3 of SEQ ID NO:4. In some embodiments, the anti-hIL-4Rα antibody or its antigen-binding fragment comprises HCVD of SEQ ID NO:1.

[0227] According to certain embodiments of this disclosure, the anti-hIL-4Rα antibody or its antigen-binding fragment comprises the light (κ) chain complementarity-determining region (LCDR) 1 of SEQ ID NO:6, the LCDR2 of SEQ ID NO:7, and the LCDR3 of SEQ ID NO:8. In some embodiments, the anti-hIL-4Rα antibody or its antigen-binding fragment comprises the LCVD of SEQ ID NO:5.

[0228] According to certain other embodiments of this disclosure, the anti-hIL-4Rα antibody or its antigen-binding fragment comprises HCDR1 of SEQ ID NO:10, HCDR2 of SEQ ID NO:11, HCDR3 of SEQ ID NO:12, LCDR1 of SEQ ID NO:14, LCDR2 of SEQ ID NO:15, and LCDR3 of SEQ ID NO:16. In some embodiments, the anti-hIL-4Rα antibody or its antigen-binding fragment comprises HCVD of SEQ ID NO:9 and LCVD of SEQ ID NO:13.

[0229] According to certain other embodiments of this disclosure, the anti-hIL-4Rα antibody or its antigen-binding fragment comprises HCDR1 of SEQ ID NO:18, HCDR2 of SEQ ID NO:19, HCDR3 of SEQ ID NO:20, LCDR1 of SEQ ID NO:22, LCDR2 of SEQ ID NO:23, and LCDR3 of SEQ ID NO:24. In some embodiments, the anti-hIL-4Rα antibody or its antigen-binding fragment comprises HCVD of SEQ ID NO:17 and LCVD of SEQ ID NO:21.

[0230] Another non-limiting exemplary antibody that can be used in practice with this disclosure is called “mAb2”. This antibody is also referred to as H4H083P in U.S. Patent No. 7,608,693. mAb2 (H4H083P) comprises the HCVR / LCVR amino acid sequence pair having SEQ ID NO:9 / 13 and the HCDR1-HCDR2-HCDR3 / LCDR1-LCDR2-LCDR3 domain represented by SEQ ID NO:10-11-12 / SEQ ID NO:14-15-16.

[0231] Another non-limiting exemplary antibody that can be used in practice with this disclosure is called “mAb3”. This antibody is also referred to as H4H095P in U.S. Patent No. 7,608,693. mAb3 (H4H095P) comprises the HCVR / LCVR amino acid sequence pair having SEQ ID NO:17 / 21 and the HCDR1-HCDR2-HCDR3 / LCDR1-LCDR2-LCDR3 domain represented by SEQ ID NO:18-19-20 / SEQ ID NO:22-23-24.

[0232] The amount of antibody or antigen-binding fragment thereof contained in the pharmaceutical formulations disclosed herein may vary depending on the specific properties required by the formulation and the specific circumstances and purpose for which the formulation is intended to be used. In some embodiments, the pharmaceutical formulation is a liquid formulation, which may contain about 100±10 mg / mL to about 200±20 mg / mL of antibody, about 110±11 mg / mL to about 190±19 mg / mL of antibody, about 120±12 mg / mL to about 180±18 mg / mL of antibody, about 130±13 mg / mL to about 170±17 mg / mL of antibody, about 140±14 mg / mL to about 160±16 mg / mL of antibody, or about 150±15 mg / mL of antibody. For example, the formulations disclosed herein may comprise an antibody or an antigen-binding fragment thereof that specifically binds to hIL-4Rα at concentrations of about 90 mg / mL, about 95 mg / mL, about 100 mg / mL, about 105 mg / mL, about 110 mg / mL, about 115 mg / mL, about 120 mg / mL, about 125 mg / mL, about 130 mg / mL, about 131 mg / mL, about 132 mg / mL, about 133 mg / mL, about 134 mg / mL, about 135 mg / mL, about 140 mg / mL, about 145 mg / mL, about 150 mg / mL, about 155 mg / mL, about 160 mg / mL, about 165 mg / mL, about 170 mg / mL, about 175 mg / mL, about 180 mg / mL, about 185 mg / mL, about 190 mg / mL, about 195 mg / mL, or about 200 mg / mL.

[0233] The pharmaceutical formulations disclosed herein comprise one or more excipients. As used herein, the term "excipient" means any non-therapeutic agent added to the formulation to provide a desired consistency, viscosity, or stabilizing effect.

[0234] In some embodiments, the pharmaceutical formulation of this disclosure comprises at least one organic cosolvent, the type and amount of which stabilizes the hIL-4Rα antibody under rough treatment (e.g., vortexing). In some embodiments, "stabilize" means preventing the formation of aggregated antibodies exceeding 2% (in moles per liter) of the total antibody volume during rough treatment. In some embodiments, rough treatment involves vortexing the solution containing the antibody and the organic cosolvent for approximately 120 minutes.

[0235] In some embodiments, the organic cosolvent is a nonionic surfactant, such as alkyl poly(ethylene oxide). Specific nonionic surfactants that may be included in the formulations disclosed herein include, for example, polysorbates, such as polysorbate 20, polysorbate 28, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 81, and polysorbate 85; poloxamers, such as poloxamer 181, poloxamer 188, and poloxamer 407; or polyethylene glycol (PEG). Polysorbate 20 is also known as TWEEN 20, dehydrated sorbitan monolaurate, and polyoxyethylene dehydrated sorbitan monolaurate. Poloxamer 181 is also known as PLURONIC F68.

[0236] The amount of organic cosolvent contained in the pharmaceutical formulations of this disclosure can vary depending on the specific properties required by the formulation and the specific circumstances and purpose for which the formulation is intended to be used. In some embodiments, the formulation may contain about 0.1% ± 0.01% to about 2% ± 0.2% of a surfactant. For example, the formulations of this disclosure may contain about 0.09%, about 0.10%, about 0.11%, about 0.12%, about 0.13%, about 0.14%, about 0.15%, about 0.16%, about 0.17%, about 0.18%, about 0.19%, about 0.20%, about 0.21%, about 0.22%, about 0.23%, about 0.24%, about 0.25%, about 0.26%, about 0.27%, about 0.28%, about 0.29%, or about 0.30% of polysorbate 20 or poloxamer 181. For example, the formulations disclosed herein may contain about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, or about 2.0% of PEG 3350.

[0237] Exemplary organic cosolvents for stabilizing hIL-4Rα antibodies include 0.2% ± 0.02% polysorbate 20, 0.2% ± 0.02% poloxamer 181, or 1% ± 0.1% PEG 3350.

[0238] The pharmaceutical formulation disclosed herein may also comprise one or more heat stabilizers, the type and amount of which stabilize the hIL-4Rα antibody under thermal stress conditions. In some embodiments, "stable" means that when the solution containing the antibody and the heat stabilizer is maintained at about 45°C for up to about 28 days, greater than about 92% of the antibody remains in its native conformation. In some embodiments, "stable" means that when the solution containing the antibody and the heat stabilizer is maintained at about 45°C for up to about 28 days, less than about 5% of the antibody aggregates.

[0239] In some embodiments, the heat stabilizer is a sugar or sugar alcohol selected from sucrose, trehalose, and mannitol, or any combination thereof, and the amount of such sugar or sugar alcohol contained in the formulation may vary depending on the specific circumstances of using the formulation and its intended use. In some embodiments, the formulation may contain about 2.5% to about 10% sugar or sugar alcohol, about 3% to about 9.5% sugar or sugar alcohol, about 3.5% to about 9% sugar or sugar alcohol, about 4% to about 8.5% sugar or sugar alcohol, about 4.5% to about 8% sugar or sugar alcohol, about 5% to about 7.5% sugar or sugar alcohol, about 5.5% to about 7% sugar or sugar alcohol, or about 6.0% to about 6.5% sugar or sugar alcohol. For example, the pharmaceutical formulations disclosed herein may contain about 2.5% ± 0.375%, about 3% ± 0.45%, about 3.5% ± 0.525%, about 4.0% ± 0.6%, about 4.5% ± 0.675%, about 5.0% ± 0.75%, about 5.5% ± 0.825%, about 6.0% ± 0.9%, about 6.5% ± 0.975%, about 7.0% ± 1.05%, about 7.5% ± 1.125%, about 8.0% ± 1.2%, 8.5% ± 1.275%, about 9.0% ± 1.35%, or about 10.0% ± 1.5% of sugars or sugar alcohols (e.g., sucrose, trehalose, or mannitol).

[0240] The pharmaceutical formulation disclosed herein may also comprise a buffer or buffer system for maintaining a stable pH and aiding in the stabilization of the hIL-4Rα antibody. In some embodiments, "stable" means that less than 3.0% ± 0.5% of the antibody aggregates when the solution containing the antibody and buffer is maintained at about 45°C for up to about 14 days. In some embodiments, "stable" means that less than 3.7% ± 0.5% of the antibody aggregates when the solution containing the antibody and buffer is maintained at about 25°C for up to about 6 months. In some embodiments, "stable" means that at least 95% ± 0.5% of the antibody remains in its native conformation when the solution containing the antibody and buffer is maintained at about 45°C for up to about 14 days, as determined by size exclusion chromatography. In some embodiments, "stable" means that at least 96% ± 0.5% of the antibody remains in its native conformation when the solution containing the antibody and buffer is maintained at about 25°C for up to about 6 months, as determined by size exclusion chromatography. In some embodiments, "stable" means that when the solution containing antibody and buffer is maintained at about 45°C for up to about 14 days, at least 62% ± 0.5% of the antibody is in a neutral conformation, as determined by cation exchange chromatography. In some embodiments, "stable" means that when the solution containing antibody and buffer is maintained at about 25°C for up to about 6 months, at least 54% ± 0.5% of the antibody is in a neutral conformation, as determined by cation exchange chromatography. The term "neutral conformation" means that the antibody fraction eluted from the ion exchange resin typically has a more "basic" peak on one side and a more "acidic" peak on the other.

[0241] The pharmaceutical formulations disclosed herein can have a pH of about 5.2 to about 6.4. For example, the formulations disclosed herein can have a pH of about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, or about 6.4. In some embodiments, the pH is about 5.3 ± 0.2, about 5.9 ± 0.2, or about 6.0 ± 0.2.

[0242] In some embodiments, the buffer solution or buffer system comprises at least one buffer having a buffering range that fully or partially overlaps with the pH range of 5.2-6.4. In one embodiment, the buffer solution or buffer system comprises two buffers, with the first buffer having an effective pH range of 3.6-5.6 and the second buffer having an effective pH range of 5.5-7.4. In one embodiment, the first buffer has a pKa of about 4.8 ± 0.3, and the second buffer has a pKa of about 6.0 ± 0.3. In some embodiments, the buffer system comprises an acetate buffer and a histidine buffer. In some embodiments, histidine is present at about 1.3-1.9 parts per part acetate (in moles). In some embodiments, histidine is present at about 1.6 ± 0.25 parts to 1 part acetate (in moles). In some embodiments, the acetate is expressed in amounts from about 2.5 mM to about 22.5 mM, from about 3.0 mM to about 22 mM, from about 3.5 mM to about 21.5 mM, from about 4.0 mM to about 21.0 mM, from about 4.5 mM to about 20.5 mM, from about 5.0 mM to about 20 mM, from about 5.5 mM to about 19.5 mM, from about 6.0 mM to about 19.0 mM, from about 6.5 mM to about 18.5 mM, from about 7.0 mM to about 18.0 mM, and from about 7.5 mM to... Concentrations of about 17.5 mM, about 8.0 mM to about 17 mM, about 8.5 mM to about 16.5 mM, about 9.0 mM to about 16.0 mM, about 9.5 mM to about 15.5 mM, about 10.0 mM to about 15.0 mM, about 10.5 mM to about 14.5 mM, about 12.5 mM ± 1.875 mM, about 11.0 mM to about 14.0 mM, about 11.5 mM to about 13.5 mM, or about 12.0 mM to about 13.0 mM are present. In some embodiments, histidine is present at concentrations of about 10 mM to about 30 mM, about 11 mM to about 29 mM, about 12 mM to about 28 mM, about 13 mM to about 27 mM, about 14 mM to about 26 mM, about 15 mM to about 25 mM, about 16 mM to about 24 mM, about 17 mM to about 23 mM, about 18 mM to about 22 mM, or about 19 mM to about 21 mM. In some embodiments, the buffer system comprises about 12.5 mM of acetate and about 20 mM of histidine at a pH of about 5.9.

[0243] The pharmaceutical formulations disclosed herein may also comprise one or more excipients for maintaining reduced viscosity or reducing the viscosity of formulations containing high concentrations of protein (e.g., typically >100 mg / ml protein). In some embodiments, the formulation comprises arginine in an amount sufficient to maintain the viscosity of the liquid formulation at less than about 35 cPoise, less than about 30 cPoise, less than about 25 cPoise, less than about 20 cPoise, less than about 15 cPoise, less than about 14 cPoise, less than about 13 cPoise, less than about 12 cPoise, less than about 10 cPoise, or less than about 9 cPoise.

[0244] In some embodiments, the pharmaceutical formulation of this disclosure contains arginine at a concentration of about 25 mM ± 3.75 mM, about 50 mM ± 7.5 mM, or about 100 mM ± 15 mM, preferably L-arginine hydrochloride. In some embodiments, the arginine is about 20 mM to about 30 mM, about 21 mM to about 29 mM, about 21.25 mM to about 28.75 mM, about 22 mM to about 28 mM, about 23 mM to about 27 mM, or about 24 mM to about 26 mM.

[0245] According to one aspect of this disclosure, the pharmaceutical formulation is a low-viscosity, generally physiologically isotonic liquid formulation comprising: (i) a human antibody (e.g., mAb1, mAb2, or mAb3 [as above]) that specifically binds to hIL-4Rα at a concentration of about 100 mg / ml or higher; (ii) a buffer system that provides adequate buffering at about 5.9 ± 0.6; (iii) a sugar, particularly used as a heat stabilizer; (iv) an organic cosolvent that protects the structural integrity of the antibody; and (v) an amino acid for maintaining manageable viscosity for subcutaneous injection.

[0246] According to one embodiment, the pharmaceutical formulation comprises: (i) a human IgG1 antibody at a concentration of about 100 mg / ml to about 200 mg / ml, which specifically binds to hIL-4Rα and comprises a substituted IGHV3-9 type heavy chain variable region and a substituted IGHV2-28 type light chain variable region (e.g., mAb1); (ii) a buffer system comprising acetate and histidine, which effectively buffers at about pH 5.9 ± 0.6; (iii) sucrose as a heat stabilizer; (iv) polysorbate as an organic cosolvent; and (v) arginine as a viscosity reducer.

[0247] According to one embodiment, the pharmaceutical formulation comprises: (i) a human IgG1 antibody at a concentration of about 150 mg / ml ± 25 mg / ml, which specifically binds to hIL-4Rα, and comprising HCDR1 of SEQ ID NO:2, HCDR2 of SEQ ID NO:3, HCDR3 of SEQ ID NO:4, LCDR1 of SEQ ID NO:6, LCDR2 of SEQ ID NO:7, and LCDR3 of SEQ ID NO:8; (ii) about 12.5 mM ± 1.9 mM of acetate and about 20 mM ± 3 mM of histidine, which are effectively buffered at about pH 5.9 ± 0.3; (iii) about 5% w / v ± 0.75% w / v of sucrose; (iv) about 0.2% w / v ± 0.03% w / v of polysorbate 20; and (v) about 25 mM ± 3.75 mM of arginine in the form of L-arginine hydrochloride.

[0248] Additional non-limiting examples of pharmaceutical formulations covered by this disclosure are set forth elsewhere herein, including the working examples given below.

[0249] The pharmaceutical formulations disclosed herein generally exhibit a high level of stability. As used herein, the term "stable" with respect to a pharmaceutical formulation means that the antibody within the pharmaceutical formulation retains an acceptable degree of chemical structure or biological function after storage under defined conditions. A formulation may be stable even if the antibody contained therein does not retain 100% of its chemical structure or biological function after a defined storage time. In some cases, retention of approximately 90%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% of the antibody's structure or function after a defined storage time may be considered "stable."

[0250] Stability can be measured, in particular, by determining the percentage of natural antibodies retained in the formulation after storage at a defined temperature for a defined time. The percentage of natural antibodies can be determined, in particular, by size exclusion chromatography (e.g., size exclusion high-performance liquid chromatography [SE-HPLC]). As used herein, the phrase “acceptable level of stability” means that at least 90% of the natural form of antibody can be detected in the formulation after storage at a given temperature for a defined time. In some embodiments, at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the natural form of antibody can be detected in the formulation after storage at a defined temperature for a defined time. The defined time for measuring stability can be at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or longer. When assessing stability, the permitted storage temperature for a pharmaceutical preparation can be any temperature from about -80°C to about 45°C, such as storage at about -30°C, about -20°C, about 0°C, about 4°C–8°C, about 5°C, about 25°C, or about 45°C. For example, if after 3 months of storage at 5°C, greater than about 90%, 95%, 96%, 97%, or 98% of natural antibodies are detected by SE-HPLC, the pharmaceutical preparation can be considered stable. If after 6 months of storage at 5°C, greater than about 90%, 95%, 96%, 97%, or 98% of natural antibodies are detected by SE-HPLC, the pharmaceutical preparation can also be considered stable. If after 9 months of storage at 5°C, greater than about 90%, 95%, 96%, 97%, or 98% of natural antibodies are detected by SE-HPLC, the pharmaceutical preparation can also be considered stable. The pharmaceutical preparation can also be considered stable if, after storage at 25°C for 3 months, greater than approximately 90%, 95%, 96%, or 97% of natural antibodies are detected by SE-HPLC. Similarly, the pharmaceutical preparation can be considered stable if, after storage at 25°C for 6 months, greater than approximately 90%, 95%, 96%, or 97% of natural antibodies are detected by SE-HPLC. Finally, the pharmaceutical preparation can be considered stable if, after storage at 25°C for 9 months, greater than approximately 90%, 95%, 96%, or 97% of natural antibodies are detected by SE-HPLC.

[0251] Stability can be measured, in particular, by determining the percentage of antibodies formed in aggregates within the formulation after storage at a defined temperature for a defined time, wherein stability is inversely proportional to the percentage of aggregates formed. The percentage of aggregated antibodies can be determined, in particular, by size exclusion chromatography (e.g., size exclusion high-performance liquid chromatography [SE-HPLC]). As used herein, the phrase “acceptable level of stability” means that after storage at a defined temperature for a defined time, up to 5% of the antibody is detected in aggregate form in the formulation. In some embodiments, acceptable level of stability means that after storage at a defined temperature for a defined time, up to about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody can be detected in aggregates within the formulation after storage at a defined temperature for a defined time. The defined time for measuring stability can be at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or longer. When assessing stability, the pharmaceutical preparation can be stored at any temperature from about -80°C to about 45°C, such as at about -30°C, about -20°C, about 0°C, about 4°C-8°C, about 5°C, about 25°C, or about 45°C. For example, if less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies in aggregated form are detected after 3 months of storage at 5°C, the pharmaceutical preparation can be considered stable. If less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies in aggregated form are detected after 6 months of storage at 5°C, the pharmaceutical preparation can also be considered stable. If less than about 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies in aggregated form are detected after 9 months of storage at 5°C, the pharmaceutical preparation can also be considered stable. The pharmaceutical preparation can also be considered stable if, after storage at 25°C for 3 months, less than approximately 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies in aggregated form are detected. Similarly, the pharmaceutical preparation can be considered stable if, after storage at 25°C for 6 months, less than approximately 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies in aggregated form are detected. Finally, the pharmaceutical preparation can be considered stable if, after storage at 25°C for 9 months, less than approximately 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies in aggregated form are detected.

[0252] Stability can be measured, in particular, by determining the percentage of antibody that migrates during ion exchange in a more acidic fraction (“acidic form”) rather than in the majority fraction (“neutral conformation”), where stability is inversely proportional to the fraction of antibody in the acidic form. While not wishing to be bound by theory, deamidation of antibodies can result in antibodies carrying a greater negative charge and thus being more acidic relative to undeamidated antibodies (see, for example, Robinson, N., Protein Deamidation, PNAS, Apr. 16, 2002, 99(8): 5283-5288). The percentage of “acidified” or “deamidized” antibodies can be determined, in particular, by ion exchange chromatography (e.g., cation exchange high-performance liquid chromatography [CEX-HPLC]). As used herein, the phrase “acceptable level of stability” means that, after storage at a defined temperature for a defined time, up to 45% of the antibody is detected in the formulation in a more acidic form. In some embodiments, acceptable stability means that after storage at a given temperature for a specified period of time, up to about 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibody in its acidic form can be detectable in the formulation. The specified period for measuring stability can be at least 2 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 18 months, at least 24 months, or longer. When assessing stability, the pharmaceutical formulation can be stored at any temperature from about -80°C to about 45°C, for example, at about -30°C, about -20°C, about 0°C, about 4°C–8°C, about 5°C, about 25°C, or about 45°C. For example, if after storage at 5°C for 3 months, less than approximately 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are in a more acidic form, the pharmaceutical preparation can be considered stable. Similarly, if after storage at 25°C for 3 months, less than approximately 18%, 17%, 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies are in a more acidic form, the pharmaceutical preparation can also be considered stable. If, after being stored at 45°C for 8 weeks, less than approximately 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of the antibodies are in a more acidic form, the pharmaceutical preparation can also be considered stable.If, after storage at 40°C for 2 weeks, less than approximately 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of antibodies in a more acidic form can be detected, the pharmaceutical preparation can also be considered stable.

[0253] Other methods can be used to assess the stability of the formulations disclosed herein, such as using differential scanning calorimetry (DSC) to determine thermal stability, using controlled stirring to determine mechanical stability, and using absorbance at about 350 nm or about 405 nm to determine solution turbidity. For example, if the OD405 of the formulation changes by less than about 0.05 (e.g., 0.04, 0.03, 0.02, 0.01, or less) after being stored at about 5°C to about 25°C for 6 or more months, the formulation disclosed herein can be considered stable.

[0254] Stability can also be assessed by measuring the biological activity or binding affinity of the antibody to its target. For example, a formulation of this disclosure can be considered stable if, after storage for a defined period of time (e.g., 1 to 12 months) at, for example, 5°C, 25°C, 45°C, etc., the anti-IL-4Rα antibody contained in the formulation binds to IL-4Rα with at least 90%, 95%, or higher affinity than the antibody before storage. Binding affinity can be determined by, for example, ELISA or plasma resonance. Biological activity can be determined by an IL-4Rα activity assay, for example, by contacting cells expressing IL-4Rα with a formulation containing an anti-IL-4Rα antibody. The binding of the antibody to such cells can be measured directly, for example, by FACS analysis. Alternatively, downstream activity of the IL-4Rα system can be measured in the presence of the antibody and an IL-4Rα agonist and compared with the activity of the IL-4Rα system in the absence of the antibody. In some embodiments, IL-4Rα can be endogenous in cells. In other embodiments, IL-4Rα can be ectopically expressed in cells.

[0255] Additional methods for assessing the stability of antibodies in formulations are shown in the examples given below.

[0256] In some embodiments, the liquid pharmaceutical formulations of this disclosure may exhibit low to moderate levels of viscosity. As used herein, “viscosity” can be “kinematic viscosity” or “absolute viscosity.” Kinematic viscosity is a measure of the resistance to flow of a fluid under gravity. When two fluids of equal volume are placed in the same capillary viscometer and allowed to flow under gravity, the viscous fluid takes longer to flow through the capillary than the less viscous fluid. For example, if one fluid takes 200 seconds to complete its flow and another takes 400 seconds, the second fluid has twice the viscosity of the first fluid on the kinematic viscosity scale. Absolute viscosity, sometimes called dynamic viscosity or simple viscosity, is the product of kinematic viscosity and fluid density (absolute viscosity = kinematic viscosity × density). The dimension of kinematic viscosity is L² / T, where L is length and T is time. Typically, kinematic viscosity is expressed in centistokes (cSt). The SI unit of kinematic viscosity is mm² / s, or 1 cSt. Absolute viscosity is expressed in centipoises (cP). The SI unit for absolute viscosity is millipascal-second (mPa-s), where 1 cP = 1 mPa-s.

[0257] As used herein, with respect to the fluid formulations of this disclosure, low levels of viscosity will exhibit an absolute viscosity of less than about 15 cPoise (cP). For example, if a fluid formulation of this disclosure exhibits an absolute viscosity of about 15 cP, about 14 cP, about 13 cP, about 12 cP, about 11 cP, about 10 cP, about 9 cP, about 8 cP, or lower when measured using standard viscosity measurement techniques, the formulation will be considered to have “low viscosity.” As used herein, with respect to the fluid formulations of this disclosure, medium levels of viscosity will exhibit an absolute viscosity between about 35 cP and about 15 cP. For example, if the fluid formulation of this disclosure exhibits an absolute viscosity of about 34 cP, about 33 cP, about 32 cP, about 31 cP, about 30 cP, about 29 cP, about 28 cP, about 27 cP, about 26 cP, about 25 cP, about 24 cP, about 23 cP, about 22 cP, about 21 cP, about 20 cP, about 19 cP, 18 cP, about 17 cP, about 16 cP, or about 15.1 cP when measured using standard viscosity measurement techniques, then the formulation will be considered to have “medium viscosity”.

[0258] As illustrated in the examples below, the inventors have made a surprising discovery: by formulating antibodies with arginine at a concentration of about 25 mM to about 100 mM, it is possible to obtain liquid formulations containing high concentrations of anti-hIL-4Rα antibodies (e.g., about 100 mg / mL to at least 200 mg / mL). Furthermore, it has been found that by adjusting the sucrose content to below about 10%, the viscosity of the formulation can be reduced to even greater degrees.

[0259] The pharmaceutical formulations disclosed herein can be contained in any container suitable for storing pharmaceutical products and other therapeutic compositions. For example, the pharmaceutical formulations can be contained in sealed and sterile plastic or glass containers with a defined volume, such as vials, ampoules, syringes, cartridges, or bottles. Different types of vials can be used to contain the formulations of this disclosure, including, for example, transparent and opaque (e.g., amber) glass or plastic vials. Similarly, any type of syringe can be used to contain or administer the pharmaceutical formulations of this disclosure.

[0260] The pharmaceutical formulations disclosed herein can be contained in either "normal tungsten" syringes or "low tungsten" syringes. As will be understood by those skilled in the art, methods of manufacturing glass syringes typically involve using a heated tungsten rod to pierce the glass, thereby forming a hole through which liquid can be drawn and expelled from the syringe. This process results in the deposition of trace amounts of tungsten on the inner surface of the syringe. Subsequent washing and other treatment steps can be used to reduce the amount of tungsten in the syringe. As used herein, the term "normal tungsten" means that the syringe contains more than 500 parts per billion (ppb) of tungsten. The term "low tungsten" means that the syringe contains less than 500 ppb of tungsten. For example, the low-tungsten injector according to this disclosure may contain less than about 490, 480, 470, 460, 450, 440, 430, 420, 410, 390, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10 ppb or less of tungsten.

[0261] Rubber plungers used in syringes and rubber stoppers used to seal vial openings can be coated to prevent contamination of the pharmaceutical contents of the syringe or vial or to maintain their stability. Therefore, according to certain embodiments, pharmaceutical formulations of this disclosure can be contained in syringes including coated plungers or in vials sealed with coated rubber stoppers. For example, the plunger or stopper can be coated with a fluorocarbon film. Examples of coated stoppers or plungers suitable for use with vials and syringes containing pharmaceutical formulations of this disclosure are mentioned, for example, in U.S. Patent Nos. 4,997,423, 5,908,686, 6,286,699, 6,645,635, and 7,226,554, the contents of which are incorporated herein by reference in their entirety. Specific exemplary coated rubber stoppers and plungers that may be used in the context of this disclosure may be traded under various names. Available commercially from West Pharmaceutical Services, Inc. (Lionville, Pa.).

[0262] According to certain embodiments of this disclosure, the pharmaceutical preparation may be contained within a low-tungsten syringe comprising a plunger coated with a fluorocarbon compound.

[0263] In one embodiment, a liquid pharmaceutical preparation containing approximately 150 mg / ml ± 15 mg / ml of anti-IL-4Rα antibody is administered subcutaneously from a pre-filled syringe in an auto-injector at a volume of approximately 1 ml ± 0.15 ml.

[0264] The pharmaceutical formulations disclosed herein are particularly useful for treating, preventing, or improving any disease or condition associated with IL-4 activity, including those mediated by IL-4Rα activation. Exemplary non-limiting diseases and conditions that can be treated or prevented by administration of the pharmaceutical formulations disclosed herein include various atopic diseases, such as atopic dermatitis, allergic conjunctivitis, allergic rhinitis, asthma, and other IgE / Th2-mediated diseases.

[0265] Therefore, this disclosure includes methods for treating, preventing, or improving any disease or condition (including any of the exemplary diseases, conditions, and illnesses described above) associated with IL-4 activity or IL-4Rα activation. Treatment methods of this disclosure include administering to a subject any formulation comprising an anti-hIL-4Rα antibody disclosed herein. The subject administering the pharmaceutical formulation may be, for example, any person or non-human animal requiring such treatment, prevention, or improvement, or any person or non-human animal that would otherwise benefit from inhibiting or attenuating IL-4 or IL-4Rα-mediated activity. For example, the subject may be an individual diagnosed with any of the aforementioned diseases or conditions or considered to be at risk of developing any of the aforementioned diseases or conditions. This disclosure also includes the use of any pharmaceutical formulation disclosed herein in the preparation of a medicament for treating, preventing, or improving any disease or condition (including any of the exemplary diseases, conditions, and illnesses described above) associated with IL-4 activity or IL-4Rα activation.

[0266] Example

[0267] The following examples are provided to give those skilled in the art a complete disclosure and description of how to prepare and use the methods and compositions of this disclosure, and are not intended to limit the scope of this disclosure. Efforts have been made to ensure the accuracy of the numerical values ​​used (e.g., amounts, temperatures, etc.), but some experimental errors and deviations should also be taken into account. Unless otherwise stated, parts are molar parts, molecular weights are average molecular weights, temperatures are in degrees Celsius, and pressures are atmospheric pressure or near atmospheric pressure.

[0268] Initial formulation development activities involved screening organic solubilizers, heat stabilizers, and buffers in liquid formulations of mAb1 (the anti-IL-4Rα antibody of this disclosure) to identify excipients compatible with the protein and enhancing its stability, while maintaining the wt. molar osmotic concentration and viscosity for subcutaneous injection. Buffer conditions were also examined to determine the optimal pH for maximum protein stability.

[0269] Example 1: Organic Co-solvent

[0270] Observations revealed that mAb1 is unstable under stirring stress. Analysis by reversed-phase high-performance liquid chromatography (RP-HPLC) and size exclusion high-performance liquid chromatography (SE-HPLC) showed protein loss and increased protein aggregates when mAb1 was vortexed at room temperature (Table 1, see “No Co-solvent” data). As measured by SE-HPLC and RP-HPLC, the addition of organic co-solvents to the mAb1 solution prevented protein degradation (Table 1). However, observations showed that the addition of some organic co-solvents reduced the thermal stability of mAb1 (Table 2). Following thermal stress, as determined by RP-HPLC, a loss of protein recovery was observed in formulations containing PEG 3350 (3%) and PEG 300 (10% and 20%) (Table 2). Furthermore, as determined by SE-HPLC, formulations containing PLURONIC F68 (poloxam 181) (0.2%), PEG 300 (10% and 20%), and propylene glycol (20%) exhibited more aggregate formation than those without cosolvents. Polysorbate 20 (0.2%) and polysorbate 80 (0.2%) provided comparable stability under stirring and thermal stress.

[0271] According to Table 1, 0.3 ml of 15 mg / ml mAb1 in 10 mM phosphate (pH 6.0) along with various organic co-solvents was vortexed in a 2 ml glass vial for approximately 120 min. Turbidity was assessed by optical density (OD) at 405 nm and reported as the relative change in OD at 405 nm compared to the starting materials. The percentage of total mAb1 recovered was determined by reversed-phase HPLC (RP-HPLC). The percentages of native and aggregated mAb1 were determined by size exclusion HPLC (SE-HPLC). The SE-HPLC results given in the “Starting Materials” results are averages of the values ​​for each formulation without vortexing.

[0272] Table 1

[0273]

[0274] According to Table 2, 0.3 ml of 15 mg / ml mAb1 in 10 mM phosphate (pH 6.0) along with various organic co-solvents were stored in 2 ml glass vials at approximately 45°C for approximately 28 days. Turbidity was assessed by optical density (OD) at 405 nm and reported as the relative change in OD at 405 nm compared to the starting materials. The percentage of recovered total mAb1 was determined by reversed-phase HPLC (RP-HPLC). The percentages of native and aggregated mAb1 were determined by size exclusion HPLC (SE-HPLC). The SE-HPLC results given in the “Starting Materials” results are averages of the values ​​for each formulation under conditions without thermal stress.

[0275] Table 2

[0276]

[0277] Example 2: Heat stabilizer

[0278] The ability of various heat stabilizers, such as sugars, amino acids, and inorganic salts, to inhibit mAb1 degradation was examined while maintaining a temperature of approximately 45°C. A summary of the heat stabilizers studied is given in Table 3. Formulations containing sucrose or trehalose exhibited the greatest stabilizing effect on mAb1 in solution when incubated at elevated temperatures (as determined by SE-HPLC). Sucrose was chosen as the stabilizer due to its history of safe use in monoclonal antibody formulations.

[0279] According to Table 3, 0.3 ml of 25 mg / ml mAb1 in 10 mM acetate (pH 5.3) along with various heat stabilizers were stored in 2 ml glass vials at approximately 45°C for approximately 28 days. Turbidity was assessed by optical density (OD) at 405 nm and reported as the relative change in OD at 405 nm compared to the starting materials. Turbidity was negligible for all samples. The percentage of total mAb1 recovered was determined by reversed-phase HPLC (RP-HPLC). The percentages of native and aggregated mAb1 were determined by size exclusion HPLC (SE-HPLC). Acidic or basic substances were defined as the sum of mAb1 peaks eluted from the cation exchange (CEX-HPLC) column whose retention times were earlier or later than the main peak, respectively. The SE-HPLC results given in the “Starting Materials” results are averages of the values ​​for each formulation under no thermal stress.

[0280] Table 3

[0281] Example 3: Buffer solution and pH

[0282] The effects of pH and buffer type on mAb1 stability were also examined. 15 mg / mL mAb1 was incubated in different buffers at pH values ​​ranging from 4.5 to 7.0. Protein stability was monitored by SE-HPLC and cation exchange HPLC (CEX-HPLC). As determined by both SE-HPLC and CEX-HPLC, maximum protein stability was observed when mAb1 was formulated in histidine buffer at pH 6.0 or acetate buffer at pH 5.3 (Tables 4 and 5). Compared to formulations containing histidine buffer, the acetate buffer system provided a wider pH stability range and a lower rate of charge variant formation (Table 5). Therefore, acetate buffer at pH 5.3 was partially selected for formulations of mAb1.

[0283] According to Table 4, 0.3 ml of 15 mg / ml mAb1 and 0.2% polysorbate 20 in various buffer solutions (10 mM) were incubated in 2 ml glass vials at approximately 45°C for approximately 14 days. Turbidity was assessed by optical density (OD) at 405 nm and reported as the relative change in OD at 405 nm compared to the starting materials. Turbidity was negligible for all samples. The percentage of total mAb1 recovered was determined by reversed-phase HPLC (RP-HPLC). The percentages of native and aggregated mAb1 were determined by size exclusion HPLC (SE-HPLC). Acidic or basic substances were defined as the sum of mAb1 peaks eluted from the cation exchange (CEX-HPLC) column whose retention times were earlier or later than the main peak, respectively. The SE-HPLC results given in the “Starting Materials” results are averages of the values ​​for each formulation under conditions without thermal stress.

[0284] Table 4

[0285]

[0286] According to Table 5, 0.3 ml of 15 mg / ml mAb1 and 0.2% polysorbate 20 in various buffer solutions (10 mM) were stored in 2 ml glass vials at approximately 45°C for approximately 14 days. Turbidity was assessed by optical density (OD) at 405 nm and reported as the relative change in OD at 405 nm compared to the starting materials. Turbidity was negligible for all samples. The percentage of total mAb1 recovered was determined by reversed-phase HPLC (RP-HPLC). The percentages of native and aggregated mAb1 were determined by size exclusion HPLC (SE-HPLC). Acidic or basic substances were defined as the sum of mAb1 peaks eluted from the cation exchange (CEX-HPLC) column whose retention times were earlier or later than the main peak, respectively. The SE-HPLC results given in the “Starting Materials” results are averages of the values ​​for each formulation under conditions without thermal stress.

[0287] Formulation development studies showed that mAb1 in solution can be deamidinated under alkaline conditions (pH 6.5). Conversely, below pH 5.0, an increased rate of formation of molecular weight variants of mAb1 was observed. Based on these data, the pH of mAb1 formulations was maintained between pH 5.6 and pH 6.2. Observations revealed that mAb1 is stable within this pH range.

[0288] Table 5

[0289]

[0290] The effects of pH and buffer type on mAb1 stability were further evaluated in formulations containing 20 mM histidine (pH 6) and 12.5 mM acetate (pH 5.3) or a combination of 20 mM histidine and 12.5 mM acetate (pH 5.9) (Table 6). Compared to the buffer systems alone, mAb1 was most stable at approximately pH 5.9 in formulations containing both histidine and acetate. The slowest aggregation rate was detected when mAb1 was formulated in this combined buffer system (SE-HPLC) (Table 6).

[0291] According to Table 6, 0.4 ml of 150 mg / ml mAb1, 10% sucrose, and 0.2% polysorbate 20 in various buffer solutions were incubated in 2 ml glass vials at approximately 45°C for approximately 14 days. Turbidity was assessed by optical density (OD) at 405 nm and reported as the relative change in OD at 405 nm compared to the starting materials. Turbidity was negligible for all samples. The percentage of total mAb1 recovered was determined by reversed-phase HPLC (RP-HPLC). The percentages of native and aggregated mAb1 were determined by size exclusion HPLC (SE-HPLC). Acidic or basic substances were defined as the sum of mAb1 peaks eluted from the cation exchange (CEX-HPLC) column whose retention times were earlier or later than the main peak, respectively. The SE-HPLC results given in the “Starting Materials” results are averages of the values ​​for each formulation under conditions without thermal stress.

[0292] Table 6

[0293] Example 4: Viscosity and Tension Management

[0294] The viscosity and tonicity (expressed as weight molar osmolality) of various excipient combinations with high concentrations of mAb1 (i.e., 150 mg / ml, 175 mg / ml, and 200 mg / ml) were evaluated. The levels of sucrose, sodium chloride, and L-arginine hydrochloride were adjusted to develop formulations containing high concentrations of mAb1 at low viscosity and physiological tonicity, enabling easy, comfortable, and rapid subcutaneous delivery of large amounts of mAb1 (Table 7). A liquid formulation (pH 5.9) (Formulation A) containing 25 mM arginine, 20 mM histidine, 12.5 mM acetate, 5% (w / v) sucrose, 0.2% (w / v) polysorbate 20, and 150 mg / mL mAb1 represents an optimized formulation with low viscosity (approximately 8.5 cPoise) and physiological isotonicity (approximately 293 mOsm / kg) while maintaining mAb1 stability.

[0295] Example 5: Characterization of Formulation A

[0296] The main degradation pathways identified during the development of mAb1 liquid formulations were the formation of aggregates, cleavage products, and charge variants. The formation of these degradation products was minimized by formulating mAb1 in a formulation containing 20 mM histidine, 12.5 mM acetate, 0.2% polysorbate 20, 5% sucrose, and 25 mM L-arginine hydrochloride (pH 5.9). The formulated 150 mg / mL mAb1 was observed to be a clear to slightly milky liquid solution, substantially free of visible particles.

[0297] The prepared mAb1 was physically and chemically stable when subjected to various stresses (incubation at 25°C and 45°C) and real-time storage conditions (5°C) (Table 8). The appearance of mAb1 was unaffected when incubated at 25°C for 3 months or stored at 5°C for 6 months. Furthermore, no effect was observed on solution pH, turbidity, or the amount of mAb1 recovered. After incubation at 25°C for 3 months, no significant degradation of the antibody was observed, as determined by SE-HPLC, while a degradation of more than 3.3% was observed, as determined by CEX-HPLC. Increased degradation was observed after incubation at 45°C for 8 weeks, as determined by both SE-HPLC and CEX-HPLC, indicating that aggregate and charge variant formation are the main degradation pathways for mAb1 antibody molecules. No degradation was observed when the prepared mAb1 antibody was stored at 5°C for 6 months.

[0298] Table 7

[0299]

[0300] According to Table 8, OD = optical density; RP-HPLC = reversed-phase high-performance liquid chromatography; SE-HPLC = size exclusion high-performance liquid chromatography; and CEX-HPLC = cation exchange high-performance liquid chromatography. Acidic or basic substances are defined as the sum of mAb1 peaks eluted from the CEX-HPLC column, whose retention times are earlier or later than the main peak, respectively.

[0301] Example 6: Container

[0302] It has been established that formulations containing mAb1 are stable during filtration sterilization. Millipore MILLIPAK filtration devices were used in the production of clinical products, while filters of the same composition (Millipore MILLEXDURAPORE) were used in the studies.

[0303] Fill a 5-mL glass vial with at least 2.5mL of 150mg / mL mAb1, 5% (w / v) sucrose, 25mM L-arginine hydrochloride, 0.2% (w / v) polysorbate 20, 12.5mM acetate, and 20mM histidine (pH 5.9). Add an excess of 0.5mL of formulation to the 5-mL vial to ensure that 2.0mL of formulation can be dispensed. This excess is not to compensate for loss of mAb1 or formulations containing mAb1 during manufacturing, degradation during manufacturing, degradation during storage (shelf life), or to extend shelf life.

[0304] Compared with storage in glass vials, the stability of the prepared mAb1 (formulation A) is not affected when stored in polypropylene tubes, polystyrene tubes, polycarbonate tubes or glass vials containing stainless steel sheets (Table 9).

[0305] Table 8

[0306]

[0307] According to Table 9, 150 mg / mL mAb1, 5% sucrose, 25 mM arginine hydrochloride, 0.2% PS-20, 20 mM histidine, and 12.5 mM acetate (pH 5.9) were incubated with various materials at 40°C for 14 days. OD = optical density; RP-HPLC = reversed-phase high-performance liquid chromatography; SE-HPLC = size exclusion high-performance liquid chromatography; and CEX-HPLC = cation exchange high-performance liquid chromatography. Turbidity is reported as the relative change in OD at 405 nm compared to the starting material. Acidic or basic substances are defined as the sum of mAb1 peaks eluted from the CEX-HPLC column, whose retention times are earlier or later than the main peak, respectively.

[0308] Table 9

[0309]

[0310] Next, exemplary pharmaceutical products, drugs, and / or pharmaceutical formulations conforming to U.S. Patent No. 9,987,500B2 are described. The term “PD-1” refers to programmed death-1 protein, a T-cell co-inhibitor, also known as CD279. The amino acid sequence of full-length PD-1 is provided in GenBank as accession number NP_005009.2 and is also referred to herein as SEQ ID NO:327. The term “PD-1” also includes protein variants of PD-1 having the amino acid sequence of SEQ ID NO:321, 322, 323, or 324. The term “PD-1” includes recombinant PD-1 or fragments thereof. The term also covers PD-1 or fragments thereof coupled to, for example, a histidine tag, mouse or human Fc, or a signaling sequence such as ROR1. For example, the term includes sequences exemplified by SEQ ID NO:323 or 324 that contain, at their C-terminus, amino acid residues 25-170 of the full-length PD-1 with the C93S variation, mouse Fc (mlgG2a) or human Fc (hIgG1). Protein variants, as exemplified by SEQ ID NO:321, contain, at their C-terminus, a histidine tag coupled to amino acid residues 25-170 of the full-length PD-1. Unless specified as originating from a non-human species, the term "PD-1" refers to human PD-1.

[0311] PD-1 is a member of the CD28 / CTLA-4 / ICOS family of T-cell co-inhibitors. PD-1 is a 288-amino acid protein with an IgV-like extracellular N-terminal domain, a transmembrane domain, and intracellular domains containing both an immunoreceptor tyrosine-based inhibitory (ITIM) motif and an immunoreceptor tyrosine-based switch (ITSM) motif (Chattopadhyay et al. 2009, Immunol. revised). The PD-1 receptor has two ligands, PD-ligand-1 (PD-L1) and PD-L2.

[0312] The term "PD-L1" refers to the ligand of the PD-1 receptor, also known as CD274 and B7H1. The amino acid sequence of full-length PD-L1 is provided in GenBank as accession number NP_054862.1 and is also referred to herein as SEQ ID NO:328. The term also encompasses PD-L1 or fragments thereof coupled to, for example, histidine tags, mouse or human Fc, or signal sequences such as ROR1. For example, the term includes sequences exemplified by SEQ ID NO:325 or 326 that contain, at their C-terminus, mouse Fc (mlgG2a) or human Fc (hIgG1) coupled to amino acid residues 19-239 of full-length PD-L1. PD-L1 is a 290-amino acid protein with an extracellular IgV-like domain, a transmembrane domain, and a highly conserved intracellular domain of approximately 30 amino acids. PD-L1 is constitutively expressed in many cell types, such as antigen-presenting cells (e.g., dendritic cells, macrophages, and B cells) and hematopoietic and non-hematopoietic cells (e.g., vascular endothelial cells, pancreatic islets, and immune-exempt sites). PD-L1 is also expressed in a variety of tumors, virus-infected cells, and autoimmune tissues, and is an integral part of the immunosuppressive environment (Ribas 2012, NEJM 366:2517-2519).

[0313] As used herein, the term “T-cell co-inhibitor” refers to ligands and / or receptors that modulate immune responses through T-cell activation or inhibition. The term “T-cell co-inhibitor,” also known as T-cell co-signaling molecules, includes, but is not limited to, lymphocyte activation gene 3 protein (LAG-3, also known as CD223), cytotoxic T-lymphocyte antigen-4 (CTLA-4), B and T lymphocyte attenuating factor (BTLA), CD-28, 2B4, LY108, T-cell immunoglobulin and mucin 3 (TIM3), T-cell immune receptor with immunoglobulin and ITIM (TIGIT; also known as VSIG9), leukocyte-associated immunoglobulin-like receptor 1 (LAIR1; also known as CD305), inducible T-cell co-stimulatory factor (ICOS; also known as CD278), T-cell activation V-domain Ig repressor (VISTA), and CD160.

[0314] As used herein, the term "Fc receptor" refers to a surface receptor protein found on immune cells (including B lymphocytes, natural killer cells, macrophages, basophils, neutrophils, and mast cells) that has binding specificity to the Fc region of an antibody. The term "Fc receptor" includes, but is not limited to, Fcγ receptors [e.g., FcγRI (CD64), FcγRIIA (CD32), FcγRIIB (CD32), FcγRIIIA (CD16a), and FcγRIIIB (CD16b)], Fcα receptors (e.g., FcαRI or CD89), and Fcε receptors [e.g., FcεRI and FcεRII (CD23)].

[0315] As used herein, the term "antibody" is intended to refer to an immunoglobulin molecule (i.e., a "complete antibody molecule") consisting of four polypeptide chains, two heavy (H) chains, and two light (L) chains linked together by disulfide bonds, as well as its multimers (e.g., IgM) or its antigen-binding fragments. Each heavy chain consists of a heavy chain variable region ("HCVR" or "VH") and a heavy chain constant region (composed of domains CH1, CH2, and CH3). Each light chain consists of a light chain variable region ("LCVR" or "VL") and a light chain constant region (CL). The VH and VL regions can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments of this disclosure, the FR of the antibody (or its antigen-binding fragment) may be identical to the human germline sequence, or it may be natural or artificially modified. The common amino acid sequence may be defined based on a side-by-side analysis of two or more CDRs.

[0316] One or more CDR residues can be substituted or omitted. These antibodies, in which one or two CDRs can be omitted for binding, have been described in the scientific literature. Padlan et al. (1995 FASEB J.9:133-139) analyzed the contact regions between antibodies and their antigens based on published crystal structures and concluded that only about one-fifth to one-third of the CDR residues actually contact the antigen. Padlan also discovered many antibodies in which one or two CDRs lack the amino acid that contacts the antigen (see also Vajdos et al. 2002 J Mol Biol 320:415-428).

[0317] CDR residues that do not contact the antigen can be identified from the Kabat CDR region located outside the Chothia CDR based on previous studies (e.g., residues H60-H65 in CDRH2 are generally not required), through molecular modeling, and / or experience. If the CDR or its residues are omitted, it is usually replaced by an amino acid occupying the corresponding position in another human antibody sequence or a shared sequence of such sequences. The position within the CDR for substitution and the amino acid to be substituted can also be selected empirically. Empirical substitutions can be conserved or non-conserved.

[0318] Compared to corresponding germline sequences, the fully human anti-PD-1 monoclonal antibodies disclosed herein may contain one or more amino acid substitutions, insertions, and / or deletions in the frame regions and / or CDR regions of the heavy chain variable domain and light chain variable domain. Such mutations can be readily identified by comparing the amino acid sequences disclosed herein with germline sequences available from, for example, public antibody sequence databases. This disclosure includes antibodies and their antigen-binding fragments derived from any amino acid sequence disclosed herein, wherein one or more amino acids within one or more frame regions and / or CDR regions are mutated to corresponding residues in the germline sequence of the derived antibody, or mutated to corresponding residues in another human germline sequence, or mutated to conserved amino acid substitutions of the corresponding germline residues (such sequence changes are collectively referred to herein as “germline mutations”). Those skilled in the art can readily generate numerous antibody and antigen-binding fragments containing one or more individual germline mutations or combinations thereof, starting from the heavy chain and light chain variable region sequences disclosed herein. In some embodiments, all frame and / or CDR residues within the VH and / or VL domains are mutated back to residues found in the original germline sequence of the derived antibody. In other embodiments, only certain residues are mutated back to the original germline sequence, such as mutated residues found only in the first 8 amino acids of FR1 or the last 8 amino acids of FR4, or mutated residues found only in CDR1, CDR2, or CDR3. In other embodiments, one or more of the frame and / or CDR residues are mutated to corresponding residues of a different germline sequence (i.e., a germline sequence different from the germline sequence of the originally derived antibody). Furthermore, the antibodies of this disclosure may contain any combination of two or more germline mutations within the frame and / or CDR regions, for example, where certain individual residues are mutated to corresponding residues of a specific germline sequence, while certain other residues different from the original germline sequence remain unchanged or are mutated to corresponding residues of a different germline sequence. Once an antibody and antigen-binding fragment containing one or more germline mutations is obtained, one or more desired properties can be readily tested, such as improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as applicable), reduced immunogenicity, etc. Antibody and antigen-binding fragments obtained in this general manner are included in this disclosure.

[0319] This disclosure also includes fully human anti-PD-1 monoclonal antibodies comprising variants of any HCVR, LCVR, and / or CDR amino acid sequences disclosed herein with one or more conserved substitutions. For example, this disclosure includes anti-PD-1 antibodies having HCVR, LCVR, and / or CDR amino acid sequences with conserved amino acid substitutions, such as 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., relative to any HCVR, LCVR, and / or CDR amino acid sequences disclosed herein.

[0320] As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human mAbs of this disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo), such as in CDRs, and particularly CDR3. However, as used herein, the term "human antibody" is not intended to include mAbs in which a CDR sequence derived from another mammalian species (e.g., mouse) has been grafted onto a human FR sequence. The term includes antibodies generated in or recombined in non-human mammals. The term is not intended to include antibodies isolated from or generated in human subjects.

[0321] As used herein, the term "recombinant" means an antibody or antigen-binding fragment thereof of this disclosure produced, expressed, isolated, or obtained by techniques or methods known in the art as recombinant DNA technologies (including, for example, DNA splicing and transgenic expression). The term also refers to antibodies expressed in or isolated from a library of recombinant human antibodies in non-human mammalian (including transgenic non-human mammalian, e.g., transgenic mouse) or cell (e.g., CHO cell) expression systems.

[0322] As used herein, the term "multispecific antigen-binding molecule" refers to a bispecific, trispecific, or multispecific antigen-binding molecule and its antigen-binding fragment. A multispecific antigen-binding molecule may be specific to different epitopes of a target polypeptide, or may contain antigen-binding domains specific to epitopes of more than one target polypeptide. A multispecific antigen-binding molecule may be a single multifunctional polypeptide, or it may be a multimeric complex of two or more polypeptides covalently or non-covalently associated with each other. The term "multispecific antigen-binding molecule" includes antibodies of this disclosure that can be linked to or co-expressed with another functional molecule (e.g., another peptide or protein). For example, an antibody or fragment thereof may be functionally linked to one or more other molecular entities (such as proteins or fragments thereof) (e.g., by chemical coupling, gene fusion, non-covalent association, or other means) to produce a bispecific or multispecific antigen-binding molecule having a second binding specificity. According to this disclosure, the term "multispecific antigen-binding molecule" also includes bispecific, trispecific, or multispecific antibodies or their antigen-binding fragments. In some embodiments, the antibody of this disclosure is functionally linked to another antibody or its antigen-binding fragment to produce a bispecific antibody with a second binding specificity. Bispecific and multispecific antibodies of this disclosure are described elsewhere in this document.

[0323] The terms "specific binding" or "specific binding to" refer to the formation of a relatively stable complex between an antibody or its antigen-binding fragment and the antigen under physiological conditions. Specific binding can be characterized by an equilibrium dissociation constant of at least about 1 × 10⁻⁸ M or less (e.g., a smaller KD indicates a tighter binding). Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, etc. As described herein, antibodies that specifically bind to PD-1 have been identified by surface plasmon resonance, for example, BIACORE. TM Furthermore, multispecific antibodies that bind to one domain of PD-1 and one or more additional antigens, or bispecific antibodies that bind to two different regions of PD-1, are still considered to be "specifically binding" antibodies as used herein.

[0324] The term "high affinity" antibody refers to those mAbs that have a binding affinity (expressed in KD) of at least 10⁻⁷ M, preferably 10⁻⁸ M, more preferably 10⁻⁹ M, even more preferably 10⁻¹⁰ M, and even more preferably 10⁻¹¹ M for PD-1, such as through surface plasmon resonance (e.g., BIACORE). TM It can be measured by solution affinity ELISA.

[0325] The terms "slow dissociation rate," "Koff," or "kd" refer to the dissociation of the antibody from PD-1 with a rate constant of 1 × 10⁻³ s⁻¹ or less, preferably 1 × 10⁻⁴ s⁻¹ or less, such as by surface plasmon resonance (e.g., BIACORE). TM As determined by ).

[0326] As used herein, the terms “antigen-binding portion” and “antigen-binding fragment” of an antibody include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. As used herein, the terms “antigen-binding fragment” or “antibody fragment” of an antibody refer to one or more fragments of an antibody that retain the ability to bind to PD-1.

[0327] In specific embodiments, the antibodies or antibody fragments of this disclosure may be conjugated to portions such as ligands or therapeutic portions (“immunoconjugates”), such as antibiotics, secondary anti-PD-1 antibodies or antibodies against another antigen, such as tumor-specific antigens, autoimmune tissue antigens, virus-infected cell antigens, Fc receptors, T-cell receptors or T-cell co-inhibitors or immunotoxins, or any other therapeutic portion that may be used to treat diseases or conditions including cancer, autoimmune diseases or chronic viral infections.

[0328] As used herein, “isolated antibody” is intended to refer to an antibody that is substantially free of other antibodies (Abs) that have different antigen specificities (e.g., an isolated antibody that specifically binds to PD-1 or fragments thereof is substantially free of Abs that specifically bind to antigens other than PD-1).

[0329] As used herein, “blocking antibody” or “neutralizing antibody” (or “antibody that neutralizes PD-1 activity” or “antagonistic antibody”) is intended to refer to an antibody whose binding to PD-1 results in the inhibition of at least one biological activity of PD-1. For example, the antibodies disclosed herein can prevent or block the binding of PD-1 to PD-L1.

[0330] As used herein, "activating antibody" or "enhancing antibody" (or "agonist antibody") is intended to refer to an antibody whose binding to PD-1 results in an increase or stimulation of at least one biological activity of PD-1. For example, the antibody disclosed herein can increase the binding of PD-1 to PD-L1.

[0331] As used herein, the term "surface plasmon resonance" refers to the ability to achieve surface plasmon resonance through, for example, the use of BIACORE. TM The system detects changes in protein concentration within a biosensor matrix to analyze optical phenomena of real-time biomolecular interactions (Pharmacia Biosensor AB, Uppsala, Sweden, and Piscataway, NJ).

[0332] As used in this article, the term "KD" is intended to refer to the equilibrium dissociation constant of a particular antibody-antigen interaction.

[0333] The term "epitope" refers to an antigenic determinant that interacts with a specific antigen-binding site (called a complementary site) in the variable region of an antibody molecule. A single antigen may have more than one epitope. Therefore, different antibodies can bind to different regions on the antigen and may have different biological effects. The term "epitope" also refers to a site on an antigen that responds to B cells and / or T cells. It also refers to a region of the antigen that is bound by an antibody. An epitope can be defined as a structural epitope or a functional epitope. Functional epitopes are typically a subset of structural epitopes and have those residues that directly contribute to the interaction affinity. Epitopes can also be conformational epitopes, i.e., composed of non-linear amino acids. In some embodiments, an epitope may include determinants that are chemically active surface groups of a molecule, such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and in some embodiments may have specific three-dimensional structural features and / or specific charge features.

[0334] When referring to nucleic acids or fragments thereof, the terms "substantially identical" or "substantially the same" mean that, when optimally aligned with appropriate nucleotide insertions or deletions in another nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 90%, and more preferably at least about 95%, 96%, 97%, 98%, or 99% of the nucleotide bases, as measured by any well-known sequence identity algorithm (such as FASTA, BLAST, or GAP), as discussed below. In some cases, a nucleic acid molecule substantially identical to a reference nucleic acid molecule may encode a polypeptide having the same or substantially similar amino acid sequence as the polypeptide encoded by the reference nucleic acid molecule.

[0335] When applied to peptides, the term "substantially similar" or "fundamentally similar" means that two peptide sequences share at least 90% sequence identity when optimally aligned (e.g., using the default GAP weights via GAP or BESTFIT procedures), and more preferably at least 95%, 98%, or 99% sequence identity. Preferably, the dissimilar residue positions differ due to conserved amino acid substitutions. A "conserved amino acid substitution" is an amino acid substitution in which one amino acid residue is replaced by another amino acid residue in a side chain (R group) having similar chemical properties (e.g., charge or hydrophobicity). Typically, conserved amino acid substitutions do not substantially alter the functional properties of the protein. In cases where two or more amino acid sequences differ from each other due to conserved substitutions, the percentage or degree of similarity can be adjusted upwards to correct for the conservatism of the substitution. Methods for making such adjustments are well known to those skilled in the art. See, for example, Pearson (1994) Methods Mol. Biol. 24:307-331, which is incorporated herein by reference. Examples of amino acid groups with side chains having similar chemical properties include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic-hydroxy side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartic acid and glutamic acid; and 7) sulfur-containing side chains: cysteine ​​and methionine. Preferred conserved amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine. Alternatively, a conserved substitution is any change with a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256:1443 45, which is incorporated herein by reference. A “moderately conservative” substitution is any change that has a non-negative value in the PAM250 log-likelihood matrix.

[0336] Sequence analysis software is commonly used to measure the sequence similarity of peptides. Protein analysis software uses similarity measurements assigned to various substitutions, deletions, and other modifications (including conserved amino acid substitutions) to match similar sequences. For example, GCG software includes programs such as GAP and BESTFIT, which can be used with default parameters to determine sequence homology or sequence identity between closely related peptides (such as homologous peptides from different biological species) or between wild-type proteins and their mutants. See, for example, GCG version 6.1. Peptide sequences can also be compared using FASTA (the program in GCG version 6.1) with default or recommended parameters. FASTA (e.g., FASTA2 and FASTA3) provides alignment of the best overlapping regions between the query sequence and the search sequence and the percentage of sequence identity (Pearson (2000) ibid.). When comparing sequences disclosed herein with databases containing large numbers of sequences from different organisms, another preferred algorithm is the computer program BLAST, especially BLASTP or TBLASTN, using default parameters. See, for example, Altschul et al. (1990) J.Mol.Biol.215:403-410 and (1997) Nucleic Acids Res.25:3389-3402, each of which is incorporated herein by reference.

[0337] The phrase “therapeutic effective amount” means the amount that produces the desired effect of its application. The exact amount will depend on the therapeutic purpose and will be determined by those skilled in the art using known techniques (see, for example, Lloyd (1999) The Art, Science and Technology of Pharmaceutical Compounding).

[0338] As used herein, the term “subject” refers to an animal, preferably a mammal, that requires improvement, prevention and / or treatment of a disease or condition (such as chronic viral infection, cancer or autoimmune disease).

[0339] As used herein, “anticancer drug” means any agent that can be used to treat cancer, including but not limited to cytotoxins and agents such as antimetabolites, alkylating agents, anthracyclines, antibiotics, antimitotic agents, procarbazine, hydroxyurea, asparaginase, corticosteroids, mitotane (O,P'-(DDD)), biologics (e.g., antibodies and interferons), and radioactive agents. As used herein, “cytotoxic agent” also refers to chemotherapeutic agents and means any agent that is harmful to cells. Examples include… (Paclitaxel), temozolomide, cytochalasin B, bacitracin D, ethidium bromide, emetine, cisplatin, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthraquinone, mitoxantrone, photomycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, as well as their analogues or homologues.

[0340] As used herein, the term "antiviral drug" refers to any drug or therapy used to treat, prevent, or reduce viral infection in a host subject. The term "antiviral drug" includes, but is not limited to, zidovudine, lamivudine, abacavir, ribavirin, lopinavir, efavirenz, cobistat, tenofovir, rilpivirine, analgesics, and corticosteroids. In the context of this disclosure, viral infection includes long-term or chronic infections caused by viruses, including but not limited to human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), human papillomavirus (HPV), lymphocytic choriomeningitis virus (LCMV), and simian immunodeficiency virus (SIV).

[0341] The antibodies and antigen-binding fragments of this disclosure specifically bind to PD-1 and modulate the interaction between PD-1 and PD-L1. Anti-PD-1 antibodies can bind to PD-1 with high or low affinity. In some embodiments, the antibodies of this disclosure can be blocking antibodies, wherein the antibody can bind to PD-1 and block the interaction between PD-1 and PD-L1. In some embodiments, the blocking antibodies of this disclosure can block the binding of PD-1 to PD-L1 and / or stimulate or enhance T cell activation. In some embodiments, blocking antibodies can be used to stimulate or enhance immune responses and / or to treat subjects with cancer or chronic viral infections. When administered to subjects in need, antibodies can alleviate chronic infection with viruses such as HIV, LCMV, or HBV in the subject. They can be used to inhibit the growth of tumor cells in the subject. They can be used alone or as adjunctive therapy with other parts or forms of treatment known in the art for the treatment of cancer or viral infections.

[0342] In other embodiments, the antibody disclosed herein may be an activating antibody, wherein the antibody can bind to PD-1 and enhance the interaction between PD-1 and PD-L1. In some embodiments, the activating antibody can enhance the binding of PD-1 to PD-L1 and / or inhibit or prevent T cell activation. The activating antibody disclosed herein may be used to suppress immune responses in subjects and / or to treat autoimmune diseases.

[0343] In some embodiments, anti-PD-1 antibodies may be multispecific antigen-binding molecules, wherein they contain a first binding specificity for PD-1 and a second binding specificity for antigens selected from a group consisting of another T-cell co-inhibitor, an autoimmune tissue antigen, a T-cell receptor, an Fc receptor, a T-cell receptor, PD-L1, and different epitopes of PD-1.

[0344] In some embodiments, the antibodies of this disclosure are obtained from mice immunized with a primary immunogen such as full-length PD-1 [see GenBank accession number NP_005009.2 (SEQ ID NO:327)] or with recombinant form of PD-1 or a modified human PD-1 fragment (SEQ ID NO:321, 323 or 324) or a modified cynomolgus monkey PD-1 fragment (SEQ ID NO:322), followed by immunization with a secondary immunogen or an immunogenic active fragment of PD-1.

[0345] The immunogen may be a biologically active and / or immunogenic fragment of PD-1 or DNA encoding its active fragment. The fragment may be derived from the N-terminal or C-terminal domain of PD-1. In some embodiments of this disclosure, the immunogen is a PD-1 fragment of amino acid residues 25-170 of SEQ ID NO:327 having a C93S variation.

[0346] Peptides can be modified, including by adding or substituting certain residues, for labeling or for conjugation to carrier molecules, such as KLHs. For example, a cysteine ​​residue can be added to the N-terminus or C-terminus of the peptide, or a linker sequence can be added to prepare a peptide for conjugation to KLHs, for example, for immunization.

[0347] The full-length amino acid sequence of full-length human PD-1 is shown in SEQ ID NO:327.

[0348] In some embodiments, fragments of the aforementioned regions or peptides extending from one or both of the N-terminus or C-terminus of the regions described herein beyond the designated region by about 5 to about 20 amino acid residues can be used to prepare antibodies that specifically bind to PD-1. In some embodiments, any combination of the aforementioned regions or fragments thereof can be used to prepare PD-1 specific antibodies. In some embodiments, any one or more of the aforementioned regions of PD-1 or fragments thereof can be used to prepare monospecific, bispecific, or multispecific antibodies.

[0349] Some of the anti-PD-1 antibodies disclosed herein are capable of binding to and neutralizing the activity of PD-1, as determined by in vitro or in vivo assays. The ability of the antibodies of this disclosure to bind to PD-1 and neutralize its activity can be measured using any standard method known to those skilled in the art, including binding assays or activity assays as described herein.

[0350] Non-limiting exemplary in vitro assays for measuring binding activity are illustrated in the examples herein. In Example 3, the binding affinity and kinetic constants of the human anti-PD-1 antibody to human PD-1 and cynomolgus monkey PD-1 were determined by surface plasmon resonance and measured on a Biacore 4000 or T200 instrument. In Examples 4 and 5, blocking assays were used to determine the ability of the anti-PD-1 antibody to block PD-1 binding to PD-L1 in vitro. In Example 6, blocking assays were used to determine cross-competition between anti-PD-1 antibodies. Example 7 describes the binding of the antibody to cells overexpressing PD-1. In Example 8, a luciferase assay was used to determine the ability of the anti-PD-1 antibody to antagonize PD-1 / PD-L1 signaling in T cells.

[0351] In some embodiments, the antibodies of this disclosure are capable of enhancing or stimulating T cell activation in vitro and in subjects with cancer or infected with a virus (such as LCMV). In some embodiments, the antibodies of this disclosure are used in combination with a second therapeutic agent (such as an antibody against a second T cell co-inhibitor) to enhance the immune response of the subject and inhibit tumor growth.

[0352] Antibodies specific to PD-1 may be free of additional labeling or portions, or they may contain N-terminal or C-terminal labels or portions. In one embodiment, the label or portion is biotin. In a binding assay, the location of the label (if present) determines the orientation of the peptide relative to the surface on which it binds. For example, if the surface is coated with avidin, a peptide containing N-terminal biotin will be oriented such that the C-terminal portion of the peptide will be away from the surface. In one embodiment, the label may be a radionuclide, a fluorescent dye, or an MRI-detectable label. In some embodiments, antibodies with such labels can be used for diagnostic assays, including imaging assays.

[0353] Antigen-binding fragments of antibodies

[0354] Unless otherwise specified, as used herein, the term "antibody" should be understood to encompass an antibody molecule (i.e., a "complete antibody molecule") comprising two immunoglobulin heavy chains and two immunoglobulin light chains, and its antigen-binding fragment. As used herein, the terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, etc., include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. As used herein, the terms "antigen-binding fragment" or "antibody fragment" of an antibody refer to one or more fragments of an antibody that retain the ability to specifically bind to PD-1. Antibody fragments may include Fab fragments, F(ab')2 fragments, Fv fragments, dAb fragments, fragments containing a CDR, or isolated CDRs. In some embodiments, the term "antigen-binding fragment" refers to a polypeptide fragment of a multispecific antigen-binding molecule. In such embodiments, the term "antigen-binding fragment" includes, for example, an extracellular domain of PD-1 that specifically binds to PD-L1. Antigen-binding fragments of antibodies can be derived from intact antibody molecules, for example, using any suitable standard techniques such as proteolytic digestion or recombinant genetic engineering techniques. These techniques involve the manipulation and expression of DNA encoding variable and (optionally) constant domains of the antibody. This DNA is known and / or readily available from, for example, commercial sources, DNA libraries (including, for example, phage antibody libraries), or can be synthesized. The DNA can be sequenced and manipulated chemically or using molecular biology techniques, for example, to arrange one or more variable and / or constant domains into suitable conformations, or to introduce codons, generate cysteine ​​residues, modify, add or delete amino acids, etc.

[0355] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues of a hypervariable region of a mimic antibody (e.g., a separated complementarity-determining region (CDR), such as a CDR3 peptide) or a constrained FR3-CDR3-FR4 peptide. Other engineered molecules, such as domain-specific antibodies, single-domain antibodies, domain-deficient antibodies, chimeric antibodies, CDR-transplanted antibodies, biantibodies, triantibodies, tetraantibodies, microantibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the term "antigen-binding fragments" as used herein.

[0356] Antigen-binding fragments of antibodies typically contain at least one variable domain. The variable domain can be of any size or amino acid composition and typically contains at least one CDR adjacent to or co-framed with one or more frame sequences. In antigen-binding fragments having a VH domain associated with a VL domain, the VH and VL domains can be positioned relative to each other in any suitable arrangement. For example, the variable region can be a dimer and contain VH-VH, VH-VL, or VL-VL dimers. Alternatively, the antigen-binding fragment of an antibody can contain a monomeric VH or VL domain.

[0357] In some embodiments, the antigen-binding fragment of the antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of the variable and constant domains that may be found within the antigen-binding fragment of the antibody of this disclosure include: (i) VH-CH1; (ii) VH-CH2; (iii) VH-CH3; (iv) VH-CH1-CH2; (v) VH-CH1-CH2-CH3; (vi) VH-CH2-CH3; (vii) VH-CL; (viii) VL-CH1; (ix) VL-CH2; (x) VL-CH3; (xi) VL-CH1-CH2; (xii) VL-CH1-CH2-CH3; (xiii) VL-CH2-CH3; and (xiv) VL-CL. In any configuration of the variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be directly connected to each other or connected via complete or partial hinge or linker regions. The hinge region may consist of at least two amino acids (e.g., 5, 10, 15, 20, 40, 60, or more), resulting in flexible or semi-flexible connections between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, the antigen-binding fragment of the antibody of this disclosure may comprise a homodimer or heterodimer (or other multimer) of any of the above-described variable and constant domain configurations that are non-covalently associated (e.g., via disulfide bonds) with each other and / or with one or more monomeric VH or VL domains.

[0358] Like intact antibody molecules, antigen-binding fragments can be monospecific or multispecific (e.g., bispecific). Multispecific antigen-binding fragments of antibodies typically contain at least two distinct variable domains, each capable of specifically binding to a single antigen or different epitopes on the same antigen. Any multispecific antibody form, including the exemplary bispecific antibody forms disclosed herein, can be adapted to the antigen-binding fragments of the antibodies disclosed herein using conventional techniques available in the art.

[0359] Preparation of human antibodies

[0360] Methods for generating human antibodies in transgenic mice are known in the art. In the context of this disclosure, any such known method can be used to prepare human antibodies that specifically bind to PD-1.

[0361] An immunogen comprising any of the following can be used to generate antibodies against PD-1. In some embodiments, the antibodies of this disclosure are obtained by immunizing mice with full-length natural PD-1 (see GenBank accession number NP_005009.2) (SEQ ID NO:327) or a recombinant PD-1 peptide. Alternatively, PD-1 or fragments thereof can be produced and modified using standard biochemical techniques (SEQ ID NO:321-324) and used as an immunogen.

[0362] In some embodiments, the immunogen may be a peptide derived from the N-terminus or C-terminus of PD-1. In one embodiment, the immunogen is an extracellular domain or IgV-like domain of PD-1. In some embodiments of this disclosure, the immunogen is a PD-1 fragment of approximately amino acid residues 25-170 of SEQ ID NO:327 having a C93S variation.

[0363] In some implementations, the immunogen may be a recombinant PD-1 peptide expressed in Escherichia coli or any other eukaryotic or mammalian cell, such as Chinese hamster ovary (CHO) cells.

[0364] In some embodiments, a fragment of the aforementioned region or a peptide extending from one or both of the N-terminus or C-terminus of the region described herein beyond the designated region by about 5 to about 20 amino acid residues can be used to prepare an antibody that specifically binds to PD-1. In some embodiments, any combination of the aforementioned region or fragments thereof can be used to prepare a PD-1 specific antibody.

[0365] use Technology (see, for example, U.S. Patent No. 6,596,541, Regeneron Pharmaceuticals), (or any other known method for generating monoclonal antibodies) initially isolates a high-affinity chimeric antibody against PD-1 having a human variable region and a mouse constant region. The technology involves generating transgenic mice with a genome containing human heavy and light chain variable regions operatively linked to endogenous mouse constant region loci, enabling the mice to produce antibodies containing human variable regions and mouse constant regions in response to antigen stimulation. DNA encoding the antibody heavy and light chain variable regions is isolated and operatively linked to DNA encoding the human heavy and light chain constant regions. The DNA is then expressed in cells capable of expressing fully human antibodies.

[0366] bioequivalent

[0367] The anti-PD-1 antibodies and antibody fragments disclosed herein encompass proteins having an amino acid sequence that differs from the amino acid sequence of the antibody but retains the ability to bind PD-1. Such variant antibodies and antibody fragments contain one or more additions, deletions, or substitutions of amino acids when compared to the parental sequence, but their biological activity is substantially equivalent to that of the antibody. Similarly, the antibody-encoding DNA sequences disclosed herein encompass sequences containing one or more additions, deletions, or substitutions of nucleotides when compared to the disclosed sequences, but which encode antibodies or antibody fragments that are substantially bioequivalent to the antibodies or antibody fragments of this disclosure.

[0368] If, for example, two antigen-binding proteins or antibodies are drug equivalents or drug substitutes, and their absorption rates and extents do not show significant differences when administered under similar experimental conditions at the same molar per liter dose (whether as a single dose or multiple doses), they are considered bioequivalent. If some antibodies are equivalent in extent of absorption but not in rate of absorption, they are considered equivalents or drug substitutes, but can still be considered bioequivalent because such differences in absorption rates are intentional and reflected in the label, are not necessary for achieving effective in vivo drug concentrations for example, with long-term use, and are considered to have no medical significance for the specific drug product under investigation.

[0369] In one implementation, two antigen-binding proteins are bioequivalent if they do not differ clinically in terms of safety, purity, or potency.

[0370] In one implementation, if a patient can switch between the reference product and the biologic one or more times without an expected increase in the risk of side effects, including clinically significant changes in immunogenicity or reduced efficacy, compared to continued treatment without such switching, the two antigen-binding proteins are bioequivalent.

[0371] In one implementation, if two antigen-binding proteins act on one or more conditions of use through one or more common mechanisms of action, they are bioequivalent, provided that such mechanisms are known.

[0372] Bioequivalence can be demonstrated by in vivo and / or in vitro methods. Bioequivalence measurements include, for example, (a) in vivo studies in humans or other mammals, in which changes in the concentration of the antibody or its metabolites in blood, plasma, serum or other biological fluids over time are measured; (b) in vitro studies that correlate with and reasonably predict in vivo bioavailability data; (c) in vivo studies in humans or other mammals, in which changes in the appropriate acute pharmacological effect of the antibody (or its target) over time are measured; and (d) in well-controlled clinical trials that establish the safety, efficacy, or bioavailability or bioequivalence of the antibody.

[0373] Bioequivalent variants of the antibodies disclosed herein can be constructed, for example, by various substitutions or deletions of biologically unwanted terminal or internal residues or sequences. For example, cysteine ​​residues that are not essential for biological activity can be deleted or replaced with other amino acids to prevent the formation of unwanted or incorrect intramolecular disulfide bonds during renaturation. In other cases, bioequivalent antibodies may include antibody variants comprising amino acid changes (e.g., mutations that eliminate or remove glycosylation) that incorporate the glycosylation characteristics of the modified antibody.

[0374] Anti-PD-1 antibodies containing Fc variants

[0375] According to certain embodiments of this disclosure, anti-PD-1 antibodies comprising an Fc domain containing one or more mutations, such mutations enhancing or reducing antibody binding to FcRn receptors, for example, at acidic pH compared to neutral pH. For example, this disclosure includes anti-PD-1 antibodies containing a mutation in the CH2 or CH3 region of the Fc domain, wherein the mutation increases the affinity of the Fc domain for FcRn in acidic environments (e.g., in endosomes with a pH range of about 5.5 to about 6.0). When administered to animals, such mutations may result in a prolonged serum half-life of the antibody. Non-limiting examples of such Fc modifications include, for example, modifications at position 250 (e.g., E or Q); 250 and 428 (e.g., L or F); 252 (e.g., L / Y / F / W or T), 254 (e.g., S or T) and 256 (e.g., S / R / Q / E / D or T); or modifications at positions 428 and / or 433 (e.g., H / L / R / S / P / Q or K) and / or 434 (e.g., A, W, H, F or Y [N434A, N434W, N434H, N434F or N434Y]); or modifications at positions 250 and / or 428; or modifications at positions 307 or 308 (e.g., 308F, V308F) and 434. In one embodiment, the modifications include: 428L (e.g., M428L) and 434S (e.g., N434S) modifications; 428L, 259I (e.g., V259I) and 308F (e.g., V308F) modifications; 433K (e.g., H433K) and 434 (e.g., 434Y) modifications; 252, 254 and 256 (e.g., 252Y, 254T and 256E) modifications; 250Q and 428L modifications (e.g., T250Q and M428L); and 307 and / or 308 modifications (e.g., 308F or 308P). In yet another embodiment, the modifications include 265A (e.g., D265A) and / or 297A (e.g., N297A) modifications.

[0376] For example, this disclosure includes an anti-PD-1 antibody comprising an Fc domain comprising one or more mutation pairs or mutation groups selected from the group consisting of: 250Q and 248L (e.g., T250Q and M248L); 252Y, 254T and 256E (e.g., M252Y, S254T and T256E); 428L and 434S (e.g., M428L and N434S). ); 257I and 311I (e.g., P257I and Q311I); 257I and 434H (e.g., P257I and N434H); 376V and 434H (e.g., D376V and N434H); 307A, 380A and 434A (e.g., T307A, E380A and N434A); and 433K and 434F (e.g., H433K and N434F). In one embodiment, this disclosure includes an anti-PD-1 antibody comprising an Fc domain containing an S108P mutation in the hinge region of IgG4 to promote dimer stability. All possible combinations of the aforementioned Fc domain mutations and other mutations within the variable domains of the antibodies disclosed herein are considered within the scope of this disclosure.

[0377] This disclosure also includes anti-PD-1 antibodies comprising a chimeric heavy chain constant (CH) region, wherein the chimeric CH region comprises segments derived from CH regions of more than one immunoglobulin isotype. For example, the antibody of this disclosure may comprise a chimeric CH region comprising part or all of a CH2 domain derived from a human IgG1, human IgG2, or human IgG4 molecule, combined with part or all of a CH3 domain derived from a human IgG1, human IgG2, or human IgG4 molecule. According to certain embodiments, the antibody of this disclosure comprises a chimeric CH region having a chimeric hinge region. For example, the chimeric hinge may comprise an “upper hinge” amino acid sequence (amino acid residues from positions 216 to 227 according to EU designation) derived from a hinge region of human IgG1, human IgG2, or human IgG4, combined with a “lower hinge” sequence (amino acid residues from positions 228 to 236 according to EU designation) derived from a hinge region of human IgG1, human IgG2, or human IgG4. According to certain embodiments, the chimeric hinge region comprises amino acid residues derived from the upper hinge of human IgG1 or human IgG4 and amino acid residues derived from the lower hinge of human IgG2. In some embodiments, antibodies comprising the chimeric CH region as described herein can exhibit modified Fc effector function without adversely affecting the therapeutic or pharmacokinetic properties of the antibody. (See, for example, U.S. Serial No. 14 / 170,166, filed January 31, 2014, the disclosure of which is incorporated herein by reference in its entirety.)

[0378] Biological characteristics of antibodies

[0379] Typically, the antibodies of this disclosure exert their effects by binding to PD-1. This disclosure includes anti-PD-1 antibodies and antigen-binding fragments thereof that bind to soluble monomeric or dimer PD-1 molecules with high affinity. For example, this disclosure includes antibodies that bind to monomeric PD-1 at less than about 50 nM KD (e.g., at 25°C or 37°C) and antigen-binding fragments of the antibodies, as measured by surface plasmon resonance, for example using an assay format as defined in Example 3 herein. In some embodiments, the antibody or its antigen-binding fragment binds to monomeric PD-1 at less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 2 nM, or less than about 1 nM KD, as measured by surface plasmon resonance, for example using an assay format as defined in Example 3 herein or a substantially similar assay.

[0380] This disclosure also includes antibodies and antigen-binding fragments thereof with KD-binding dimer PD-1 at less than about 400 pM (e.g., at 25°C or 37°C), as measured by surface plasmon resonance, for example using an assay format as defined in Example 3 herein. In some embodiments, antibodies or antigen-binding fragments thereof with KD-binding dimer PD-1 at less than about 300 pM, less than about 250 pM, less than about 200 pM, less than about 100 pM, or less than about 50 pM, as measured by surface plasmon resonance, for example using an assay format as defined in Example 3 herein or a substantially similar assay.

[0381] This disclosure also includes antibodies or antigen-binding fragments thereof that bind to cynomolgus monkey (Macaca fascicularis) PD-1 at a KD concentration of less than about 35 nM (e.g., at 25°C or 37°C), as measured by surface plasmon resonance, for example using an assay format as defined in Example 3 herein. In some embodiments, antibodies or antigen-binding fragments thereof bind to cynomolgus monkey PD-1 at KD concentrations of less than about 30 nM, less than about 20 nM, less than about 15 nM, less than about 10 nM, or less than about 5 nM, as measured by surface plasmon resonance, for example using an assay format as defined in Example 3 herein or a substantially similar assay.

[0382] This disclosure also includes antibodies and antigen-binding fragments of PD-1 with a dissociation half-life (t1 / 2) greater than about 1.1 minutes, as measured by surface plasmon resonance at 25°C or 37°C, for example using an assay format defined as in Example 3 herein or a substantially similar assay. In some embodiments, the antibodies or antigen-binding fragments of this disclosure have a t1 / 2 time of greater than about 5 minutes, greater than about 10 minutes, greater than about 30 minutes, greater than about 50 minutes, greater than about 60 minutes, greater than about 70 minutes, greater than about 80 minutes, greater than about 90 minutes, greater than about 100 minutes, greater than about 200 minutes, greater than about 300 minutes, greater than about 400 minutes, greater than about 500 minutes, greater than about 600 minutes, greater than about 700 minutes, greater than about 800 minutes, greater than about 900 minutes, greater than about 1000 minutes, or greater than about 1200 minutes. 1 / 2 Combined with PD-1, such as by surface plasmon resonance at 25°C or 37°C, for example using an assay format as defined in Example 3 of this document (e.g., mAb capture or antigen capture format) or substantially similar assay.

[0383] This disclosure also includes antibodies or antigen-binding fragments thereof that block the binding of PD-1 to PD-L1 with an IC50 of less than about 3 nM, as determined by an ELISA-based immunoassay (e.g., as shown in Example 4) or substantially similar assays. This disclosure also includes antibodies and antigen-binding fragments thereof that bind to PD-1 and enhance the binding of PD-1 to PD-L1.

[0384] In some embodiments, the antibody of this disclosure can bind to an extracellular domain of PD-1 or a fragment of that domain. In some embodiments, the antibody of this disclosure can bind to more than one domain (cross-reactive antibody). In some embodiments, the antibody of this disclosure can bind to an epitope located in an extracellular domain comprising amino acid residues 21-171 of PD-1 (SEQ ID NO: 327). In one embodiment, the antibody can bind to an epitope comprising one or more amino acids selected from the group consisting of amino acid residues 1-146 of SEQ ID NO: 321-324.

[0385] In some embodiments, the antibodies of this disclosure can exert their effects by binding to any other region or fragment of the full-length protein, the amino acid sequence of which is shown in SEQ ID NO:327, to block or inhibit PD-1-associated PD-L1 binding activity. In some embodiments, the antibodies can attenuate or modulate the interaction between PD-1 and PD-L1.

[0386] In some embodiments, the antibody disclosed herein may be a bispecific antibody. The bispecific antibody of this disclosure may bind to one epitope in one domain of PD-1 and may also bind to a second epitope in a different domain of PD-1. In some embodiments, the bispecific antibody of this disclosure may bind to two different epitopes in the same domain. In one embodiment, the multispecific antigen-binding molecule comprises: a first binding specificity, wherein the first binding specificity comprises an extracellular domain of PD-L1 or a fragment thereof; and a second binding specificity for another epitope of PD-1.

[0387] In one embodiment, this disclosure provides an isolated fully human monoclonal antibody or antigen-binding fragment thereof that binds to PD-1, wherein the antibody or fragment thereof exhibits one or more of the following characteristics: (i) comprising an amino acid sequence having an amino acid sequence selected from the group consisting of SEQ ID NO:2, 18, 34, 50, 66, 82, 98, 114, 130, 146, 162, 178, 194, 210, 218, 226, 234, 242, 250, 258, 266, 274, 282, 290, 298, 306, and 314, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; (ii) comprising an HCVR having an amino acid sequence selected from the group consisting of SEQ ID NO:2, 18, 34, 50, 66, 82, 98, 114, 130, 146, 162, 178, 194, 210, 218, 226, 234, 242, 250, 258, 266, 274, 282, 290, 298, 306, and 314; (ii) comprising an amino acid sequence having an amino acid sequence selected from the group consisting of SEQ ID NO:2, 18, 34, 50, 66, 82, 98, 114, 130, 146, 162, 178, 194, 210, 218, 226, 234, 242, 250, 258, 266, 274, 28 LCVR comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 138, 154, 170, 186, and 202, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; (iii) an HCDR3 domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 8, 24, 40, 56, 72, 88, 104, 120, 136, 152, 168, 184, 200, 216, 224, 232, 240, 248, 256, 264, 272, 280, 288, 296, 304, 312, and 320, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; and a LCVR ... HCDR3 domain having an amino acid sequence selected from the group consisting of SEQ ID (iv) An LCDR3 domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 16, 32, 48, 64, 80, 96, 112, 128, 144, 160, 176, 192 and 208, or a substantially similar sequence having at least 90%, at least 95%, at least 98% or at least 99% sequence identity;The HCDR2 domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 6, 22, 38, 54, 70, 86, 102, 118, 134, 150, 166, 182, 198, 214, 222, 230, 238, 246, 254, 262, 270, 278, 286, 294, 302, 310, and 318, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; the LCDR1 domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 12, 28, 44, 60, 76, 92, 108, 124, 140, 156, 172, 188, and 204, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; and the HCDR2 domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 6, 22, 38, 54, 70, 86, 102, 118, 134, 150, 166, 182, 198, 214, 222, 230, 238, 246, 254, 262, 270, 278, 286, 294, 302, 310, and 318, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; and the HCDR2 domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 6 NO: The amino acid sequence of the group consisting of 14, 30, 46, 62, 78, 94, 110, 126, 142, 158, 174, 190 and 206 or substantially similar sequences having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; (v) is a multispecific antigen-binding molecule containing a first binding specificity for PD-1 and a second binding specificity for antigens selected from the group consisting of PD-1, tumor-specific antigens, autoimmune tissue-specific antigens, virus-infected cell antigens, different T-cell co-inhibitors, T-cell receptors and Fc receptors; (vi) with (vii) Binding of human PD-1 with KD at approximately 28 pM to approximately 1.5 μM; (viii) Binding of cynomolgus monkey PD-1 with KD at approximately 3 nM to approximately 7.5 μM; (viii) Blocking or enhancing the binding of PD-1 to PD-L1 with an IC50 of ≤ approximately 3.3 nM; (ix) Blocking PD-1-induced T cell downregulation and / or rescuing T cell signaling in a T cell / APC luciferase reporter assay; (x) Stimulating T cell proliferation and activity in a mixed lymphocyte response (MLR) assay; (xi) Inducing IL-2 and / or IFNγ production in an MLR assay; and (xii) Inhibiting tumor growth and prolonging survival in subjects with cancer.

[0388] In one embodiment, this disclosure provides an isolated fully human monoclonal antibody or antigen-binding fragment thereof that blocks the binding of PD-1 to PD-L1, wherein the antibody or fragment thereof exhibits one or more of the following features: (i) comprising an HCVR having an amino acid sequence selected from the group consisting of SEQ ID NO: 130, 162, 234 and 314 or a substantially similar sequence having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; (ii) comprising an LCVR having an amino acid sequence selected from the group consisting of SEQ ID NO: 138, 170, 186 and 202 or a substantially similar sequence having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; and having an HCVR having an amino acid sequence selected from the group consisting of SEQ ID NO: 138, 170, 186 and 202 or a substantially similar sequence having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; and having an HCVR having an amino acid sequence selected from the group consisting of SEQ ID NO: 136, 168, 240 and 320 ...0, 162, 234 and 314 or a substantially similar sequence having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; and having an (iv) An LCDR3 domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 144, 176, 192, and 208, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; an HCDR1 domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 132, 164, 236, and 316, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; an HCDR2 domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 134, 166, 238, and 318, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; an LCDR1 domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 140, 172, 188, and 204, or a substantially similar sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity; and an LCDR3 domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 14 ...; and an LCDR3 NO: The group consisting of amino acid sequences of NO: 142, 174, 190 and 206 or substantially similar sequences having at least 90%, at least 95%, at least 98% or at least 99% sequence identity; (v) is a multispecific antigen-binding molecule having a first binding specificity for PD-1 and a second binding specificity for antigens selected from the group consisting of different epitopes of PD-1, tumor-specific antigens, autoimmune tissue-specific antigens, viral-infected cell antigens, different T-cell co-inhibitors, T-cell receptors and Fc receptors; (vi) binds human PD-1 with ≤10⁻⁹ M KD; (vii) binds cynomolgus monkey PD-1 with 10⁻⁸ M KD;(viii) Blocking the binding of PD-1 to PD-L1 with an IC50 of ≤10⁻¹⁰ M; (ix) Blocking PD-1-induced T cell downregulation and / or rescuing T cell signaling in a T cell / APC luciferase reporter assay; (x) Stimulating T cell proliferation and activity in a mixed lymphocyte response (MLR) assay; (xi) Inducing IL-2 and / or IFNγ production in an MLR assay; and (xii) Inhibiting tumor growth and prolonging survival in subjects with cancer.

[0389] The antibodies disclosed herein may have one or more of the foregoing biological characteristics or any combination thereof. Other biological characteristics of the antibodies disclosed herein will be apparent to those skilled in the art from this overview of the disclosure (including the working examples herein).

[0390] Species selectivity and species cross-reactivity

[0391] According to certain embodiments of this disclosure, the anti-PD-1 antibody binds to human PD-1 but not to PD-1 from other species. Alternatively, in some embodiments, the anti-PD-1 antibody of this disclosure binds to human PD-1 and PD-1 from one or more non-human species. For example, the anti-PD-1 antibody of this disclosure may bind to human PD-1 and may, depending on the circumstances, bind to or not bind to one or more of mouse, rat, guinea pig, hamster, gerbil, pig, cat, dog, rabbit, goat, sheep, cattle, horse, camel, cynomolgus monkey, marmoset, rhesus monkey, or chimpanzee PD-1. In some embodiments, the anti-PD-1 antibody of this disclosure may bind to human and cynomolgus monkey PD-1 with the same or different affinities but not to rat and mouse PD-1.

[0392] Epitope plotting and related techniques

[0393] This disclosure includes anti-PD-1 antibodies that interact with one or more amino acids found within one or more domains of a PD-1 molecule, the domains including, for example, extracellular (IgV-like) domains, transmembrane domains, and intracellular domains containing immunoreceptor tyrosine-based inhibitory motifs (ITIM) and immunoreceptor tyrosine-based switching motifs (ITSM). The antibody-binding epitope may consist of a single, continuous sequence of three or more amino acids (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more) located within any of the aforementioned domains of the PD-1 molecule (e.g., a linear epitope within the domain). Alternatively, the epitope may consist of multiple non-continuous amino acids (or amino acid sequences) located within one or both of the aforementioned domains of the PD-1 molecule (e.g., a conformational epitope).

[0394] Various techniques known to those skilled in the art can be used to determine whether an antibody “interacts with one or more amino acids within a peptide or protein.” Exemplary techniques include, for example, conventional cross-blocking assays, such as those described in Antibodies, Harlow, and Lane (Cold Spring Harbor Press, Cold Spring Harbor, NY). Other methods include alanine scanning mutation analysis, peptide blotting analysis (Reineke (2004) Methods Mol. Biol. 248:443-63), peptide cleavage analysis, crystallographic studies, and NMR analysis. Additionally, methods such as epitope excision, epitope extraction, and chemical modification of the antigen can be employed (Tomer (2000) Prot. Sci. 9:487-496). Another method that can be used to identify amino acids within a peptide that interact with the antibody is hydrogen / deuterium exchange detected by mass spectrometry. Generally, hydrogen / deuterium exchange methods involve deuterating the protein of interest and then binding the antibody to the deuterated protein. Next, the protein / antibody complex is transferred to water, and the exchangeable protons within the amino acids protected by the antibody complex undergo deuterium-to-hydrogen reverse exchange at a slower rate than those within amino acids not part of the interface. Therefore, amino acids forming part of the protein / antibody interface may retain deuterium and thus exhibit relatively higher quality compared to amino acids not included in the interface. After antibody dissociation, the target protein is subjected to protease cleavage and mass spectrometry analysis to reveal deuterium-tagged residues corresponding to the specific amino acids interacting with the antibody. See, for example, Ehring (1999) Analytical Biochemistry 267:252-259; Engen and Smith (2001) Anal. Chem. 73:256A-265A.

[0395] The term "epitope" refers to a site on an antigen that responds to B cells and / or T cells. B cell epitopes can be formed from consecutive or non-consecutive amino acids arranged side-by-side through the ternary folding of a protein. Epitopes formed from consecutive amino acids are generally retained upon exposure to denaturing solvents, while epitopes formed through ternary folding are generally lost upon treatment with denaturing solvents. Epitopes typically contain at least three, and more often at least five, or eight to ten amino acids in a unique spatial conformation.

[0396] Modification-assisted analysis (MAP), also known as antigen structure-based antibody analysis (ASAP), is a method for classifying a large number of monoclonal antibodies (mAbs) against the same antigen based on the similarity of their binding profiles to chemically or enzymatically modified antigen surfaces (see US 2004 / 0101920, which is incorporated herein by reference in its entirety). Each category can reflect a unique epitope that is distinctly different from or partially overlaps with an epitope represented by another category. This technique allows for rapid filtering of genetically identical antibodies, enabling characterization to focus on genetically distinct antibodies. When applied to hybridoma screening, MAP can facilitate the identification of rare hybridoma clones that produce mAbs with the desired characteristics. MAP can be used to sort the antibodies of this disclosure into groups of antibodies that bind to different epitopes.

[0397] In some embodiments, an anti-PD-1 antibody or its antigen-binding fragment binds to epitopes in any one or more regions of PD-1, either in its natural form (as illustrated in SEQ ID NO: 327) or recombinant form (as illustrated in SEQ ID NO: 321-324), or to fragments thereof. In some embodiments, the antibody binding of this disclosure comprises an extracellular region of one or more amino acids selected from the group consisting of amino acid residues 21-171 of PD-1. In some embodiments, the antibody binding of this disclosure comprises an extracellular region of one or more amino acids selected from the group consisting of amino acid residues 1-146 (as illustrated in SEQ ID NO: 322) of cynomolgus monkey PD-1.

[0398] In some embodiments, as shown in Table 1, the antibody of this disclosure interacts with at least one amino acid sequence selected from the group consisting of amino acid residues from about position 21 to about position 136 of SEQ ID NO:327 or amino acid residues from about position 136 to about position 171 of SEQ ID NO:327. These regions are partially exemplified in SEQ ID NO:321-324.

[0399] This disclosure includes anti-PD-1 antibodies that bind to the same epitope or a portion of an epitope as any particular exemplary antibody described in Table 1 herein or an antibody having a CDR sequence of any exemplary antibody described in Table 1. Similarly, this disclosure also includes anti-PD-1 antibodies that compete with any particular exemplary antibody described in Table 1 herein or an antibody having a CDR sequence of any exemplary antibody described in Table 1 for binding to PD-1 or a PD-1 fragment. For example, this disclosure includes anti-PD-1 antibodies that cross-competitively bind to PD-1 with one or more antibodies (e.g., H2aM7788N, H4×H8992P, H4×H8999P, H1M7799N, H2aM7780N, H1M7800N, H2aM7794N, H2aM7798N, H4×H9145P2, H4H9057P2, H4×H9120P2, H4×H9128P2, H4H9019P, H4×H9119P2, H4×H9135P2, H4×H9034P, H2aM7790N, H4×H9035P, H4×H9037P, H4×H9045P, and H2aM7795N) as defined in Example 6 herein.

[0400] Using conventional methods known in the art, it can be readily determined whether an antibody binds to the same epitope as or competes with a reference anti-PD-1 antibody. For example, to determine whether a test antibody binds to the same epitope as a reference anti-PD-1 antibody of this disclosure, the reference antibody is allowed to bind to the PD-1 protein or peptide under saturation conditions. Next, the ability of the test antibody to bind to the PD-1 molecule is evaluated. If the test antibody is able to bind to PD-1 after saturation binding with the reference anti-PD-1 antibody, it can be concluded that the test antibody binds to a different epitope than the reference anti-PD-1 antibody. On the other hand, if the test antibody cannot bind to the PD-1 protein after saturation binding with the reference anti-PD-1 antibody, the test antibody can bind to the same epitope as the one bound by the reference anti-PD-1 antibody of this disclosure.

[0401] To determine whether an antibody competitively binds to a reference anti-PD-1 antibody, the binding method described above is performed in two directions: In the first direction, the reference antibody is allowed to bind to the PD-1 protein under saturation conditions, and then the binding of the test antibody to the PD-1 molecule is evaluated. In the second direction, the test antibody is allowed to bind to the PD-1 molecule under saturation conditions, and then the binding of the reference antibody to the PD-1 molecule is evaluated. If, in both directions, only the first (saturated) antibody is able to bind to the PD-1 molecule, it is concluded that the test antibody and the reference antibody compete for binding to PD-1. As will be understood by those skilled in the art, the antibody competing for binding with the reference antibody may not necessarily bind to the same epitope as the reference antibody, but the binding of the reference antibody can be spatially blocked by binding to overlapping or adjacent epitopes.

[0402] If two antibodies competitively inhibit (block) the binding of another antibody to an antigen, then the two antibodies bind to the same or overlapping epitopes. That is, an excess of 1, 5, 10, 20, or 100 times that of one antibody inhibits the binding of the other antibody by at least 50%, but preferably 75%, 90%, or even 99%, as measured in a competitive binding assay (see, for example, Junghans et al., Cancer Res. 1990 50:1495-1502). Alternatively, if substantially all amino acid mutations in the antigen that reduce or eliminate the binding of one antibody reduce or eliminate the binding of the other antibody, then the two antibodies have the same epitope. If some amino acid mutations that reduce or eliminate the binding of one antibody reduce or eliminate the binding of the other antibody, then the two antibodies have overlapping epitopes.

[0403] Additional routine experiments (e.g., peptide mutation and binding assays) can then be performed to confirm whether the observed lack of binding to the test antibody is actually due to binding to the same epitope as the reference antibody or whether steric occlusion (or another phenomenon) is the cause of the observed lack of binding. Such experiments can be performed using ELISA, RIA, surface plasmon resonance, flow cytometry, or any other quantitative or qualitative antibody binding assay available in the art.

[0404] Immunoconjugates

[0405] This disclosure covers human anti-PD-1 monoclonal antibodies conjugated to a therapeutic portion (“immunoconjugate”) (such as a cytotoxin or chemotherapeutic agent) to treat cancer. As used herein, the term “immunoconjugate” refers to an antibody chemically or biologically linked to a cytotoxin, radiopharmaceutical, cytokine, interferon, target or reporter portion, enzyme, toxin, peptide, or protein or therapeutic agent. An antibody may be linked to a cytotoxin, radiopharmaceutical, cytokine, interferon, target or reporter portion, enzyme, toxin, peptide, or therapeutic agent at any position along the molecule, as long as it can bind to its target. Examples of immunoconjugates include antibody-pharmaceutical conjugates and antibody-toxin fusion proteins. In one embodiment, the agent may be a second, different antibody targeting PD-1. In some embodiments, the antibody may be conjugated to an agent specific to tumor cells or virus-infected cells. The type of therapeutic portion that may be conjugated to an anti-PD-1 antibody will take into account the condition to be treated and the desired therapeutic effect to be achieved. Examples of suitable agents for forming immunoconjugates are known in the art; see, for example, WO 05 / 103081.

[0406] Multispecific antibodies

[0407] The antibodies disclosed herein can be monospecific, bispecific, or multispecific antibodies. Multispecific antibodies can be specific to different epitopes of a single target polypeptide, or may contain antigen-binding domains specific to more than one target polypeptide. See, for example, Tutt et al., 1991, J. Immunol. 147:60-69; Kufer et al., 2004, Trends Biotechnol. 22:238-244.

[0408] In one aspect, this disclosure includes a multispecific antigen-binding molecule or an antigen-binding fragment thereof, wherein one specificity of the immunoglobulin is specific to the extracellular domain of PD-1 or a fragment thereof, while another specificity of the immunoglobulin is specific to binding outside the extracellular domain of PD-1 or to a second therapeutic target, or conjugated to a therapeutic portion. In some embodiments, the first antigen-binding specificity may comprise PD-L1 or PD-L2 or a fragment thereof. In some embodiments of this disclosure, one specificity of the immunoglobulin is specific to an epitope comprising amino acid residues 21-171 of PD-1 (SEQ ID NO:327) or a fragment thereof, while another specificity of the immunoglobulin is specific to a second target antigen. The second target antigen may be on the same cells as PD-1 or on different cells. In one embodiment, the second target cell is on immune cells other than T cells, such as B cells, antigen-presenting cells, monocytes, macrophages, or dendritic cells. In some embodiments, the second target antigen may be present on tumor cells or autoimmune tissue cells or virus-infected cells.

[0409] In another aspect, this disclosure provides a multispecific antigen-binding molecule or an antigen-binding fragment thereof, comprising a first antigen-binding specificity for binding PD-1 and a second antigen-binding specificity for binding T-cell receptors, B-cell receptors, or Fc receptors. In a related aspect, this disclosure provides a multispecific antigen-binding molecule or an antigen-binding fragment thereof, comprising a first antigen-binding specificity for binding PD-1 and a second antigen-binding specificity for binding various T-cell co-inhibitors (such as LAG-3, CTLA-4, BTLA, CD-28, 2B4, LY108, TIGIT, TIM3, LAIR1, ICOS, and CD160).

[0410] On the other hand, this disclosure provides a multispecific antigen-binding molecule or an antigen-binding fragment thereof, comprising a first antigen-binding specificity for binding PD-1 and a second antigen-binding specificity for binding autoimmune tissue-specific antigens. In some embodiments, the antibody may be an activating antibody or an agonist antibody.

[0411] Any multispecific antigen-binding molecule or variant thereof disclosed herein may be constructed using standard molecular biology techniques (e.g., recombinant DNA and protein expression techniques) as known to those skilled in the art.

[0412] In some implementations, PD-1-specific antibodies are generated in a bispecific form (“bispecific antibody”), wherein variable regions that bind to different domains of PD-1 are linked together to confer bidomain specificity within a single binding molecule. Properly designed bispecific antibodies can enhance overall PD-1 inhibitory efficacy by increasing specificity and binding affinity. Variable regions specific to individual domains (e.g., segments of the N-terminal domain) or variable regions capable of binding to different regions within a domain are paired on a structural scaffold that allows each region to simultaneously bind a separate epitope or to different regions within a domain. In one example of a bispecific antibody, a heavy chain variable region (VH) from a binder specific to one domain is recombined with light chain variable regions (VL) from a series of binders specific to a second domain to identify non-homologous VL partners that can pair with the original VH without disrupting the original specificity of that VH. In this way, a single VL segment (e.g., VL1) can be combined with two different VH domains (e.g., VH1 and VH2) to produce bispecific antibodies consisting of two binding "arms" (VH1-VL1 and VH2-VL1). The use of a single VL segment reduces the complexity of the system, thereby simplifying and improving the efficiency of the cloning, expression, and purification processes used to generate bispecific antibodies (see, for example, US Serial Nos. 13 / 022,759 and US2010 / 0331527).

[0413] Alternatively, antibodies binding more than one domain and a second target (such as, but not limited to, a second different anti-PD-1 antibody) can be prepared in a bispecific form using the techniques described herein or other techniques known to those skilled in the art. Antibody variable regions binding different regions can be linked together with variable regions binding, for example, to relevant sites on the extracellular domain of PD-1 to confer dual antigen specificity within a single binding molecule. Appropriately designed bispecific antibodies of this nature have dual functions. Variable regions specific for the extracellular domain are combined with variable regions specific for the exterior of the extracellular domain, and paired on a structural scaffold that allows each variable region to bind to a separate antigen.

[0414] Exemplary bispecific antibody forms that can be used within the context of this disclosure involve the use of a first immunoglobulin (Ig) CH3 domain and a second Ig CH3 domain, wherein the first and second Ig CH3 domains differ from each other by at least one amino acid, and wherein the at least one amino acid difference reduces the binding of the bispecific antibody to protein A compared to a bispecific antibody lacking this amino acid difference. In one embodiment, the first Ig CH3 domain binds to protein A, and the second Ig CH3 domain contains a mutation that reduces or eliminates protein A binding, such as an H95R modification (exon numbered according to IMGT; H435R according to EU). The second CH3 may also contain a Y96F modification (Y436F according to IMGT; Y436F according to EU). Other modifications that may be found in the second CH3 include: in the case of IgG1 antibodies, D16E, L18M, N44S, K52N, V57M, and V82I (based on IMGT; D356E, L358M, N384S, K392N, V397M, and V422I); in the case of IgG2 antibodies, N44S, K52N, and V82I (based on IMGT; N384S, K392N, and V422I); and in the case of IgG4 antibodies, Q15R, N44S, K52N, V57M, R69K, E79Q, and V82I (based on IMGT; Q355R, N384S, K392N, V397M, R409K, E419Q, and V422I). Variations of the above-described bispecific antibody forms are considered within the scope of this disclosure.

[0415] Other exemplary bispecific forms that may be used in the context of this disclosure include, but are not limited to, scFv-based or dual-antibody bispecific forms, IgG-scFv fusion proteins, dual variable domain (DVD)-Ig, Quadroma, mortar structure, common light chain (e.g., common light chain with mortar structure, etc.), CrossMab, CrossFab, (SEED) body, leucine zipper, Duobody, IgG1 / IgG2, dual-action Fab (DAF)-IgG, and Mab2 bispecific forms (for a review of the above forms, see, for example, Klein et al. 2012, mAbs 4:6, 1-11 and the references cited therein). Bispecific antibodies can also be constructed using peptide / nucleic acid conjugations, for example, where non-natural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugations, which then self-assemble into multimeric complexes having defined composition, valence, and geometry. (See, for example, Kazane et al., J. Am. Chem. Soc. [Epub: December 4, 2012]).

[0416] Therapeutic application and formulation

[0417] This disclosure provides therapeutic compositions comprising the anti-PD-1 antibody or its antigen-binding fragment thereof. The therapeutic compositions according to this disclosure will be administered together with suitable carriers, excipients, and other pharmaceutical agents incorporated into the formulation to provide improved transfer, delivery, tolerability, etc. Many suitable formulations can be found in all formularies known to medicinal chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, lipid-containing (cationic or anionic) vesicles (such as LIPOFECTIN). TM DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, polyethylene glycol emulsions (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing polyethylene glycol. See also Powell et al., “Compendium of excipients for parenteral formulations”, PDA (1998), J Pharm SciTechnol 52:238-311.

[0418] The dosage of the antibody can vary depending on the age and body size of the subject to be administered, the target disease, the condition, the route of administration, etc. When the antibody of this disclosure is used to treat or prevent a disease in an adult patient, it is generally advantageous to administer the antibody of this disclosure in a single dose of about 0.1 to about 60 mg / kg body weight, more preferably about 5 to about 60 mg / kg body weight, about 10 to about 50 mg / kg body weight, or about 20 to about 50 mg / kg body weight. The frequency and duration of treatment can be adjusted according to the severity of the condition. In some embodiments, the antibody of this disclosure or its antigen-binding fragment may be administered at an initial dose of at least about 0.1 mg to about 800 mg, about 1 to about 500 mg, about 5 to about 300 mg, or about 10 to about 200 mg, about 100 mg, or about 50 mg. In some embodiments, a second or more subsequent doses of the antibody or its antigen-binding fragment may be administered after the initial dose, in amounts that may be substantially the same as or less than the initial dose, wherein the intervals between subsequent doses are at least 1 to 3 days; at least one week; at least 2 weeks; at least 3 weeks; at least 4 weeks; at least 5 weeks; at least 6 weeks; at least 7 weeks; at least 8 weeks; at least 9 weeks; at least 10 weeks; at least 12 weeks; or at least 14 weeks.

[0419] Various delivery systems are known and can be used to administer the pharmaceutical compositions disclosed herein, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, and receptor-mediated endocytosis (see, for example, Wu et al. (1987) J. Biol. Chem. 262:4429-4432). Methods of administration include, but are not limited to, intradermal, transdermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered via any convenient route, such as by infusion or bolus, absorption through epithelial or mucosal skin linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and can be administered together with other bioactive agents. Administration can be systemic or local. The pharmaceutical compositions can also be delivered in vesicles (particularly liposomes) (see, for example, Langer (1990) Science 249:1527-1533).

[0420] This document also considers the use of nanoparticles to deliver the antibodies disclosed herein. Antibody-conjugated nanoparticles can be used for both therapeutic and diagnostic applications. Antibody-conjugated nanoparticles and methods for their preparation and use are described in detail in M. et al. 2009 (“Antibody-conjugated nanoparticles for biomedical applications” in J. Nanomat. LV2009, Article ID 439389, p. 24, doi:10.1155 / 2009 / 439389), which is incorporated herein by reference. Nanoparticles can be developed and conjugated with antibodies contained in pharmaceutical compositions to target tumor cells, autoimmune tissue cells, or virus-infected cells. Nanoparticles for drug delivery have also been described, for example, in U.S. Patent Nos. 8,257,740 or 8,246,995, each of which is incorporated herein by reference in its entirety.

[0421] In some cases, the drug composition can be delivered using a controlled-release system. In one embodiment, a pump can be used. In another embodiment, a polymer material can be used. In yet another embodiment, the controlled-release system can be placed near the target of the composition, thus requiring only a portion of the systemic dose.

[0422] Injectable formulations can include dosage forms for intravenous, subcutaneous, intradermal, intracranial, intraperitoneal, and intramuscular injection, infusion, etc. These injectable formulations can be prepared by methods known to the public. For example, injectable formulations can be prepared, for instance, by dissolving, suspending, or emulsifying the aforementioned antibodies or their salts in a sterile aqueous or oily medium conventionally used for injection. As an aqueous medium for injection, examples include physiological saline, isotonic solutions containing glucose and other adjuvants, which can be used in combination with suitable solubilizers such as alcohols (e.g., ethanol), polyols (e.g., propylene glycol, polyethylene glycol), and nonionic surfactants [e.g., polysorbate 80, HCO-50 (a polyoxyethylene (50 mol) adduct of hydrogenated castor oil)]. As an oily medium, examples include sesame oil and soybean oil, which can be used in combination with solubilizers such as benzyl benzoate, benzyl alcohol, etc. The prepared injection is preferably packaged in a suitable ampoule.

[0423] The pharmaceutical compositions disclosed herein can be delivered subcutaneously or intravenously via an autoinjector, as previously described herein. Such an autoinjector can be reusable or disposable. Reusable autoinjectors typically utilize a replaceable cartridge containing the pharmaceutical composition. Once all the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The autoinjector can then be reused. Disposable autoinjectors do not have replaceable cartridges. Instead, the autoinjector is pre-filled with the pharmaceutical composition held in a reservoir within the device. Once the pharmaceutical composition in the reservoir is emptied, the entire device is discarded.

[0424] The amount of antibody contained is typically about 5 to about 500 mg per unit dose; particularly in injectable form, the antibody content is preferably about 5 to about 100 mg, and for other dosage forms, the antibody content is about 10 to about 250 mg.

[0425] Therapeutic uses of antibodies

[0426] The antibodies disclosed herein are particularly useful for treating, preventing, and / or improving any disease or condition associated with or mediated by PD-1 expression, signaling, or activity, or for treating by blocking the interaction between PD-1 and PD-1 ligands (e.g., PD-L1 or PD-L2) or otherwise inhibiting PD-1 activity and / or signaling. For example, this disclosure provides methods for treating cancer (tumor growth inhibition), chronic viral infections, and / or autoimmune diseases by administering an anti-PD-1 antibody (or a pharmaceutical composition comprising an anti-PD-1 antibody) as described herein to a patient in need of such treatment. The antibodies of this disclosure can be used to treat, prevent, and / or improve diseases or conditions such as cancer, autoimmune diseases, or viral infections, and / or to improve at least one symptom associated with such disease, condition, or illness. In the context of the treatments described herein, anti-PD-1 antibodies can be administered as a monotherapy (i.e., as the sole therapeutic agent) or in combination with one or more additional therapeutic agents (examples of which are described elsewhere herein).

[0427] In some embodiments of this disclosure, the antibodies described herein may be used to treat subjects with primary or recurrent cancer, including but not limited to renal cell carcinoma, colorectal cancer, non-small cell lung cancer, brain cancer (e.g., glioblastoma multiforme), head and neck squamous cell carcinoma, gastric cancer, prostate cancer, ovarian cancer, kidney cancer, breast cancer, multiple myeloma, and melanoma.

[0428] These antibodies can be used to treat early or late-stage symptoms of cancer. In one embodiment, the antibodies or fragments thereof disclosed herein can be used to treat metastatic cancer. These antibodies can be used to reduce, inhibit, or shrink tumor growth in both solid tumors and hematologic malignancies. In some embodiments, treatment with the antibodies or antigen-binding fragments thereof disclosed herein results in tumor regression of more than 50%, more than 60%, more than 70%, more than 80%, or more than 90% in a subject. In some embodiments, the antibodies can be used to prevent tumor recurrence. In some embodiments, the antibodies can be used to prolong the overall survival of subjects with cancer. In some embodiments, the antibodies can be used to reduce toxicity caused by chemotherapy or radiation therapy while maintaining long-term survival in patients with cancer.

[0429] In some embodiments, the antibodies of this disclosure can be used to treat subjects with chronic viral infections. In some embodiments, the antibodies of this disclosure can be used to reduce viral titers in the host and / or rescue exhausted T cells. In some embodiments, the antibodies of this disclosure or fragments thereof can be used to treat chronic viral infections caused by lymphocytic choriomeningitis virus (LCMV). In some embodiments, the antibodies of this disclosure or antigen-binding fragments thereof can be administered at therapeutic doses to patients with human immunodeficiency virus (HIV), human papillomavirus (HPV), or hepatitis B / C virus (HBV / HCV) infection. In related embodiments, the antibodies of this disclosure or antigen-binding fragments thereof can be used to treat simian immunodeficiency virus (SIV) infection in simian subjects (such as cynomolgus monkeys).

[0430] In some implementations, the blocking antibody of this disclosure can be administered to subjects with cancer or viral infection at a therapeutically effective amount.

[0431] In some embodiments, the antibodies of this disclosure can be used to treat autoimmune diseases, including but not limited to alopecia areata, autoimmune hepatitis, celiac disease, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, hemolytic anemia, inflammatory bowel disease, inflammatory myopathy, multiple sclerosis, primary biliary cirrhosis, psoriasis, rheumatoid arthritis, scleroderma, Sjögren's syndrome, systemic lupus erythematosus, vitiligo, autoimmune pancreatitis, autoimmune urticaria, autoimmune thrombocytopenic purpura, Crohn's disease, type I diabetes, eosinophilic fasciitis, eosinophilic gastroenteritis, Goodpasseerator syndrome, myasthenia gravis, psoriatic arthritis, rheumatic fever, ulcerative colitis, vasculitis, and Wegener's granulomatosis. In some embodiments, the activated antibodies of this disclosure can be used to treat subjects with autoimmune diseases.

[0432] One or more antibodies disclosed herein may be administered to alleviate or prevent one or more symptoms or conditions of a disease or illness, or to reduce its severity.

[0433] This article also considers the prophylactic use of one or more antibodies of this disclosure in patients at risk of developing diseases or conditions such as cancer, autoimmune diseases, and chronic viral infections.

[0434] In another embodiment of this disclosure, the antibody of the present invention is used to prepare a pharmaceutical composition for treating patients suffering from cancer, autoimmune diseases, or viral infections. In another embodiment of this disclosure, the antibody of the present invention is used as an adjunct therapy with any other agent or therapy known to those skilled in the art for the treatment of cancer, autoimmune diseases, or viral infections.

[0435] Combination therapies and formulations

[0436] Combination therapy may include the anti-PD-1 antibody of this disclosure and any additional therapeutic agent that may be advantageously combined with the antibody of this disclosure or with a bioactive fragment of the antibody of this disclosure.

[0437] The antibodies disclosed herein can be synergistically combined with one or more anticancer drugs or therapies used to treat cancers including, for example, renal cell carcinoma, colorectal cancer, glioblastoma multiforme, head and neck squamous cell carcinoma, non-small cell lung cancer, colon cancer, ovarian cancer, adenocarcinoma, prostate cancer, glioma, and melanoma. This document considers using the anti-PD-1 antibodies of this disclosure in combination with immunostimulatory and / or immunosupportive therapies to inhibit tumor growth and / or prolong the survival of cancer patients. Immunostimulatory therapies include direct immunostimulatory therapies, which enhance immune cell activity by “relieving the suppression of suppressed immune cells” or “accelerating” the activation of an immune response. Examples include targeting other checkpoint receptors, vaccination, and adjuvants. Forms of immunosupportive therapy can increase the antigenicity of tumors by promoting immunogenic cell death, inflammation, or other indirect effects that promote antitumor immune responses. Examples include radiation, chemotherapy, anti-angiogenic agents, and surgery.

[0438] In various embodiments, one or more antibodies of this disclosure may be used in combination with the following agents: antibodies against PD-L1, secondary antibodies against PD-1 (e.g., nivolumab), LAG-3 inhibitors, CTLA-4 inhibitors (e.g., ipilimumab), TIM3 inhibitors, BTLA inhibitors, TIGIT inhibitors, CD47 inhibitors, another T-cell co-inhibitor or ligand antagonist (e.g., antibodies against CD-28, 2B4, LY108, LAIR1, ICOS, CD160, or VISTA), and indoleamine-2,3-dioxygenase (IDO) inhibitors. Anti-VEGF agents, vascular endothelial growth factor (VEGF) antagonists [e.g., "VEGF-Trap", such as aflibercept or other VEGF-inhibiting fusion proteins as described in U.S. Patent No. 7,087,411, or anti-VEGF antibodies or their antigen-binding fragments (e.g., bevacizumab or ranibizumab) or small molecule kinase inhibitors of the VEGF receptor (e.g., sunitinib, sorafenib, or pazopanib)], Ang2 inhibitors (e.g., nesivamab), transforming growth factor β (TGFβ) inhibitors, epidermal growth factor receptor (EGFR) inhibitors (e.g., erlotinib, cetuximab), co-stimulatory agents, etc. Agonists of stimulatory receptors (e.g., glucocorticoid-induced TNFR-related protein agonists), antibodies against tumor-specific antigens (e.g., CA9, CA125, melanoma-associated antigen 3 (MAGE3), carcinoembryonic antigen (CEA), vimentin, tumor-M2-PK, prostate-specific antigen (PSA), mucin-1, MART-1, and CA19-9), vaccines (e.g., BCG, a cancer vaccine), adjuvants that enhance antigen presentation (e.g., granulocyte-macrophage colony-stimulating factor), and bispecific antibodies (e.g., CD3×CD20 bispecific antibody, PSMA×...). CD3 bispecific antibodies), cytotoxins, chemotherapeutic agents (e.g., dacarbazine, temozolomide, cyclophosphamide, docetaxel, doxorubicin, daunorubicin, cisplatin, carboplatin, gemcitabine, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, and vincristine), cyclophosphamide, radiation therapy, IL-6R inhibitors (e.g., sarilumab), IL-4R inhibitors (e.g., dupilumab), IL-10 inhibitors, cytokines such as IL-2, IL-7, IL-21, and IL-15, antibody-drug conjugates (ADCs) (e.g., anti-CD19-DM4 ADC and anti-DS6-DM4 ADC), anti-inflammatory drugs (e.g., corticosteroids and nonsteroidal anti-inflammatory drugs), dietary supplements such as antioxidants, or any palliative therapy used to treat cancer. In some embodiments, the anti-PD-1 antibody disclosed herein can be used in combination with cancer vaccines (including dendritic cell vaccines, oncolytic viruses, tumor cell vaccines, etc.) to enhance the anti-tumor response.Examples of cancer vaccines that can be used in combination with the anti-PD-1 antibodies disclosed herein include the MAGE3 vaccine for melanoma and bladder cancer, the MUC1 vaccine for breast cancer, and EGFRv3 (e.g., Rindopepimut) or ALVAC-CEA (for CEA+ cancers) for brain cancers (including glioblastoma multiforme).

[0439] In some embodiments, the anti-PD-1 antibody of this disclosure may be administered in combination with radiotherapy in methods for producing a long-lasting anti-tumor response and / or prolonging the survival of cancer patients. In some embodiments, the anti-PD-1 antibody of this disclosure may be administered before, simultaneously with, or after radiotherapy to a cancer patient. For example, radiotherapy may be administered to the tumor lesion in one or more doses, followed by administration of one or more doses of the anti-PD-1 antibody of this disclosure. In some embodiments, radiotherapy may be locally administered to the tumor lesion to enhance the local immunogenicity of the patient's tumor (adjuvant radiation) and / or kill tumor cells (ablation radiation), followed by systemic administration of the anti-PD-1 antibody of this disclosure. For example, intracranial radiation to a patient with brain cancer (e.g., glioblastoma multiforme) may be combined with systemic administration of the anti-PD-1 antibody of this disclosure. In some embodiments, the anti-PD-1 antibody of this disclosure may be administered in combination with radiotherapy and a chemotherapeutic agent (e.g., temozolomide) or a VEGF antagonist (e.g., aflibercept).

[0440] In some embodiments, the anti-PD-1 antibody of this disclosure may be administered in combination with one or more antiviral drugs to treat chronic viral infections caused by LCMV, HIV, HPV, HBV, or HCV. Examples of antiviral drugs include, but are not limited to, zidovudine, lamivudine, abacavir, ribavirin, lopinavir, efavirenz, cobistat, tenofovir, rilpivirine, and corticosteroids. In some embodiments, the anti-PD-1 antibody of this disclosure may be administered in combination with any antagonist of a LAG3 inhibitor, a CTLA-4 inhibitor, or another T-cell co-inhibitor to treat chronic viral infections.

[0441] In some embodiments, the anti-PD-1 antibody of this disclosure can be combined with antibodies targeting Fc receptors on immune cells for the treatment of autoimmune diseases. In one embodiment, the antibody of this disclosure or a fragment thereof is administered in combination with an antibody or antigen-binding protein targeting an autoimmune tissue-specific antigen. In some embodiments, the antibody of this disclosure or an antigen-binding fragment thereof is administered in combination with an antibody or antigen-binding protein targeting T cell receptors or B cell receptors (including, but not limited to, Fcα (e.g., CD89), Fcγ (e.g., CD64, CD32, CD16a, and CD16b), CD19, etc.). The antibodies of fragments disclosed herein can be used in combination with any drugs or therapies known in the art (e.g., corticosteroids and other immunosuppressants) to treat autoimmune diseases or conditions, including but not limited to alopecia areata, a...

Claims

1. An injection device, comprising: The needle is capable of moving from a retracted configuration to a deployed configuration; vials containing one or more of dupilumab and cimiprimab; A piston configured to move within the vial; A motor configured to apply force to the piston; and A controller, connected to the motor, wherein the controller is configured to: Receive an instruction that the injection device is positioned to contact the user; and Upon receiving the instruction, a signal is sent to the motor to apply force to the piston in a first direction, wherein the force applied in the first direction causes (1) the needle to move from the retracted configuration to the deployed configuration, and (2) the medicine to be dispensed from the vial through the needle.

2. The injection device of claim 1, wherein the controller is further configured to: Upon receiving the instruction and after sending the signal to apply force to the motor in the first direction, a signal is automatically sent to the motor to apply force to the piston in the second direction to retract the needle, without any user intervention.

3. The injection device of claim 1, further comprising a housing that encloses the vial, the piston, the motor, the controller, and the needle when the needle is in the retracted configuration, wherein the needle extends out of the housing in the deployment configuration.

4. The injection device of claim 1, further comprising a cover or shield, the cover or shield including the distal portion of the needle in the retracted configuration.

5. The injection device of claim 1, further comprising an audio module, a vision module, and a tactile module, each of which is connected to the controller and configured to provide feedback to the user of the injection device.

6. The injection device of claim 1, further comprising a top that seals the opening of the vial, the top including a portion comprising a rubber material permeable to a sterilizing agent, wherein the needle includes a proximal portion configured to engage with the vial, and the proximal portion of the needle is disposed within the portion formed of the rubber material prior to fluid communication between the needle and the vial.

7. The injection device of claim 1, further comprising a cantilever coupled to the controller and movable by the needle, wherein, When the needle is in the retracted configuration, the cantilever forms part of an open circuit that sends a signal to the controller that the needle is in the retracted configuration, and when the needle is in the deployed configuration, the cantilever forms part of a closed circuit that sends a signal to the controller that the needle is in the deployed configuration.

8. The injection device of claim 1, further comprising one or more spring contacts configured to form an electrical connection between the motor and the controller.

9. The injection device of claim 1, further comprising an activation switch movable between extended positioning and pressed positioning; The indication is at least in part based on the activation switch being in the pressed position.

10. The injection device of claim 1, further comprising a touch sensor configured to detect contact with skin. The indication is at least in part based on the touch sensor detecting contact with the skin.

11. The injection device of claim 10, wherein the touch sensor includes a capacitive sensing electrode and is configured to detect contact with skin based on a change in the capacitance of the capacitive sensing electrode.

12. The injection device of claim 2, further comprising: A transmitter configured to transmit light through the vial; and A detector configured to receive the light and detect interruptions to the light by the piston.

13. The injection device of claim 12, wherein the controller is further configured to send the signal to the motor in response to an interruption of the light by the piston to apply force to the piston in the second direction.

14. The injection device of claim 12, wherein the controller is further configured to send the signal to the motor to apply force to the piston in the second direction in response to an interruption of the light by the piston after a predetermined delay.

15. The injection device of claim 1, further comprising a wireless communication module configured to transmit one or more of the following information to a remote device: diagnostic information of the injection device, information indicating an error state of the injection device, and information indicating that the injection is complete.

16. The injection device of claim 1, further comprising a wireless communication module configured to receive an activation command from a remote device, wherein the indication is at least in part based on the activation command.

17. The injection device of claim 1, wherein the vial is configured to be in fluid communication with the needle.

18. The injection device of claim 1, wherein in response to the motor driving the piston in the first direction, the needle establishes fluid communication with the vial.

19. An injection device, comprising: A housing having a tissue-joining surface and an opening in the tissue-joining surface; The needle is capable of moving from a retracted configuration to a deployed configuration through the opening; An activation switch is configured to move from a deactivated configuration to an activated configuration; vials configured to form fluid communication with the needle, wherein the vials contain one or more of dupilumab and cimipril; A motor configured to move the needle through the opening; and A controller, connected to the motor, wherein the controller is configured to: In response to a trigger, the motor moves the needle from the retracted configuration to the deployed configuration and dispenses the medicine from the vial through the needle; The triggering includes receiving a first indication that the activation switch has been moved from the deactivated configuration to the activated configuration.

20. The injection device of claim 19, further comprising: A touch sensor, which is disposed on the tissue-attaching surface and configured to detect contact with the skin; The trigger also includes a second indication that the touch sensor is in contact with the skin.

21. The injection device of claim 19, further comprising: A lock removable from the housing, the lock having a first part and a second part, wherein: In the first configuration where the lock is connected to the housing, the first portion of the lock is disposed outside the housing, and the second portion of the lock is disposed inside the housing, between the container and the conduit; In the first configuration, the second portion of the lock prevents the conduit from moving into fluid communication with the fluid contained in the container; and In the second configuration, where the lock is removed from the injection device, the conduit is movable to be in fluid communication with the fluid contained in the container.

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