Chemical dehydration and suspension delivery of protein drug products

Abstract

Disclosed herein are processes for dehydrating antibody-containing formulations, processes of forming a high concentration antibody suspension, dehydrated antibody-containing formulations prepared according to the processes described herein, suspensions, including high concentration antibody suspensions, and dosage forms.

Description

CHEMICAL DEHYDRATION AND SUSPENSION DELIVERY OF PROTEIN DRUGPRODUCTSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority of U.S. Provisional Patent Application No. 63 / 624,112, filed on January' 23, 2024, the disclosure of which is incorporated by reference herein.TECHNICAL FIELD

[0002] The disclosure provides processes for dehydrating antibody-containing formulations and uses for the dehydrated antibody-containing formulations.BACKGROUND

[0003] Monoclonal antibodies (mAb) have become a critical class of therapeutics for their high affinity', specificity', and PK profile. Intravenous mAb administration allows the delivery of enough volume of low concentration mAb (e.g., 25-150 mg / mL) to meet the dose requirements with maximum bioavailability'. Subcutaneous mAb administration may improve patient compliance and reduce the cost to hospitals and clinics, but in order to achieve subcutaneous delivery. mAbs must be formulated at higher concentrations to reduce the delivery volume while achieving the same dose requirements. At high concentrations (e.g., over 100 mg / mL), however, mAb formulation are susceptible to high viscosity7, aggregation, microbead flocculation, stability concerns, suspension stability in PFS (prefilled syringes), and issues with settling, among others.SUMMARY

[0004] Disclosed herein are processes for dehydrating an antibody-containing formulation. The processes comprise introducing the antibody-containing formulation into a dehydrating solvent mixture comprising 1 -pentanol and a cosolvent that is an ester, wherein the v / v ratio of the 1 -pentanol to the cosolvent is about 70% of 1 -pentanol to about 90% of 1- pentanol: about 30% of cosolvent to about 10% of cosolvent, to thereby dehydrate the antibody-containing formulation. In some embodiments, the ester is ethyl lactate.

[0005] Dehydrated antibody-containing formulations prepared according to the processes described herein are also disclosed.

[0006] Suspensions prepared according to the processes described herein are further disclosed

[0007] Also disclosed herein are dosage forms comprising the suspensions described herein.

[0008] Further disclosed herein are suspensions comprising a dehydrating solvent comprising 1-pentanol and a cosolvent that is an ester, wherein the v / v ratio of the 1 -pentanol to the cosolvent is about 70 to about 90: about 30 to about 10, and an antibody-containing formulation.

[0009] Also disclosed are compositions comprising a vehicle and a plurality of antibody-containing microbeads, wherein the plurality of antibody-containing microbeads comprise at least about 150 mg / mL of an antibody and wherein the composition has one or more of the following characteristics as compared to a control composition comprising antibody-containing microbeads formed using 100% 1-pentanol: an injection force of less than about 45 N when suspended in benzyl benzoate and injected through a 27 G manual prefilled syringe (PFS); a reduced density compared to the control composition as exemplified in FIG. 9; a reduced percentage of flocculation as compared to the control composition as exemplified in FIGs. 3 and 4; or about 10% or less separation for up to about 2 hours as compared to the control composition as exemplified in FIG. 9.

[0010] Also disclosed are compositions comprising a vehicle and a plurality of antibody-containing microbeads, wherein the composition has one or more of the following characteristics as compared to a control composition comprising antibody -containing microbeads formed using 100% 1-pentanol: an injection force of about 2 to about 20 N when sieved with a 75 pm sieve, suspended in benzy l benzoate, and injected through a 27 G manual prefilled syringe (PFS); about 60% total weight of the material being < 75 microns (usable material) as compared to about 85% total weight of the material being > 75 microns (unusable material) when formed using 100% 1-pentanol; a reduced settling of particles (about 80 % weight of particles formed using 100% 1-pentanol are >75 micron and settle in about 2 minutes or less as compared to about 40 % weight of particles formed using ethyl lactate and 1-pentanol which are >75 micron;a reduced setling (higher sedimentation rate) compared to a control composition as exemplified in FIG. 9; a greater particle uniformity as compared to particles formed using 100% 1- pentanol (75-micron sieved powder showed an increase of greater than 50% in particles with fractures and flocculations when dehydrated with 100% 1- pentanol alone; a greater powder uniformity as compared to particles formed using 100% 1- pentanol (94% of particles in a 75-micron sieved powder were within a 30 pm range as compared to 87% of particles in a 75-micron sieved powder formed using 100% 1-pentanol alone); smaller particle sizes as compared to particles formed using 100% 1-pentanol; or smaller 75-micron sieved particle sizes as compared to particles formed using100% 1-pentanol (particle sizes of 15 urn as compared to particle sizes of 25 urn when formed using 100% 1-pentanol); a reduced percentage of flocculation as compared to the control composition as exemplified in FIGs. 3 and 4.

[0011] Further disclosed are processes for forming a high concentration antibody suspension. The processes comprise dehydrating an antibody-containing formulation as described herein to form a dehydrated antibody-containing formulation; and resuspending the dehydrated antibody-containing formulation in a vehicle to form the high concentration antibody suspension.BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The summary, as well as the following detailed description, is further understood when read in conjunction with the appended drawings. For the purpose of illustrating the disclosed processes, dehydrated antibody-containing formulations, suspensions, dosage forms, and compositions, there are shown in the drawings exemplary embodiments of the processes, dehydrated antibody-containing formulations, suspensions, dosage forms, and compositions; however, the processes, dehydrated antibody-containing formulations, suspensions, dosage forms, and compositions are not limited to the specific embodiments disclosed. In the drawings:

[0013] FIG. 1A, FIG. IB, FIG. 1C, FIG. ID, FIG. IE, FIG. IF, FIG. 1G, and FIG. 1H are microscopy images of the chemically dehydrated microbeads using the noted solvents and an exemplary anti-PD- 1 Ab. Images were obtained at 40X magnification of theanti-PD-1 Ab microbeads suspended in benzyl benzoate. The scale at bottom of each image is 100 microns.

[0014] FIG. 2A, FIG. 2B, FIG. 2C, and FIG. 2D are microscopy images of exemplary anti-PD-1 Ab microbeads in different % ethyl lactate / 1 -pentanol solvent mixtures. Images of anti-PD-1 Ab microbeads produced by 10% (FIG. 2A), 20% (FIG. 2B), 30% (FIG. 2C), and 40% (FIG. 2D) ethyl lactate / 1 -pentanol organic solvent mixtures and suspended in benzyl benzoate are shown. Images were taken at 40X magnification and the scale at bottom of each image is 100 microns.

[0015] FIG. 3A, FIG. 3B, FIG. 3C, FIG. 3D, FIG. 3E, FIG. 3F, and FIG. 3G are photographs of solids produced using varying flow-rates in 1 -pentanol or different dehydrating solvent combinations at a selected flow rate using an exemplary anti -TL la Ab. The chemically dehydrated anti-TLla Ab in 100% 1-pentanol utilized flow rate of 50 pL / min (FIG. 3A), 25 pL / min (FIG. 3B), or 10 pL / min (FIG. 3C). Chemically dehydrated anti- TLla Ab utilized a flow rate of 25 pL / min in 1-pentanol and 30% ethyl lactate (FIG. 3D), 20% ethyl lactate (FIG. 3E), 30% ethyl lactate / 10% heptyl acetate (FIG. 3F), or 30% ethyl lactate / 10% benzyl acetate (FIG. 3G).

[0016] FIG. 4A, FIG. 4B, FIG. 4C, FIG. 4D, FIG. 4E, FIG. 4F, FIG. 4G, and FIG. 4H are images showing the evaluation of organic solvents on the size and morphology7of the chemically dehydrated antibody microbeads using an exemplary anti-TLla Ab. Microscopy of dehydrated anti-TLla Ab utilizing 100% 1-pentanol at 5X (FIG. 4A) and 40X (FIG. 4B) magnification, 30% ethyl lactate at 5X (FIG. 4C) and 40X (FIG. 4D) magnification, 30% ethyl lactate / 10% heptyd acetate at 5X (FIG. 4E) and 40X (FIG. 4F) magnification, or 30% ethyl lactate 1 10% benzyl acetate 5X (FIG. 4G) and 40X (FIG. 4H) magnification are shown. Dehydrated mAb was suspended in benzyl benzoate in order to obtain the microscopy images.

[0017] FIG. 5A, FIG. 5B, FIG. 5C, FIG. 5D, FIG. 5E, FIG. 5F, FIG. 5G, and FIG. 5H show the morphological characterization of the chemical dehydration process at a larger scale for various mAbs at 5X (FIG. 5A) and 40X (FIG. 5E) magnification, exemplary7anti-CGRP Ab at 5X (FIG. 5B) and 40X (FIG. 5F) magnification, and exemplary anti-PAR2 Ab at 5X (FIG. 5C) and 40X (FIG. 5G) magnification. Antibodies were dehydrated in 30% ethyl lactate I 70% 1-pentanol. Microscopy of exemplary anti-TLla Ab chemically dehydrated in 100% 1-pentanol at 5X (FIG. 5D) and 40X (FIG. 5H) magnification.

[0018] FIG. 6A, FIG. 6B, and FIG. 6C are line graphs of the secondary structure data of three different chemically dehydrated and reconstituted mAbs. CD overlays of aqueous stock mAh (circles) and dehydrated I reconstituted mAb (triangles).

[0019] FIG. 7A, FIG. 7B, and FIG. 7C are line graphs showing the thermal stability' of three different chemically dehydrated and reconstituted mAbs. The differential scanning calorimetry (DSC) overlays aqueous stock mAb (circles) and dehydrated / reconstituted mAb (triangles). The summary’ of DSC data is shown in Table 14.

[0020] FIG. 8 is an SEC chromatogram overlay showing the effect of dilution with yvater or buffer before chemical dehydrated of an exemplary anti-CGRP Ab after reconstitution. The SEC chromatogram overlay includes data for anti-CGRP Ab aqueous stock (control), stock diluted with buffer then dehydrated with 30% ethyl lactate / 70% 1- pentanol, and stock diluted with WFI water then dehydrated with 30% ethyl lactate.

[0021] FIG. 9A, FIG. 9B, FIG. 9C, FIG. 9D, FIG. 9E, FIG. 9F, and FIG. 9G are the visual evaluations of the suspensions in prefilled syringes (PFSs). FIG. 9A shows prepared suspensions of (1) an exemplary anti-CGRP Ab. (3) an exemplary anti-PAR2 Ab, (5) an exemplary anti-TLla Ab dehydrated with 30% ethyl lactate / 70% 1-pentanol, and (10. 11) an exemplary anti-TLla Ab dehydrated with 100% 1-pentanol. FIG. 9B shows PFS containing 250 mg / mL mAb suspension of (1) anti-CGRP Ab, (3) anti-PAR2 Ab, and (5) anti-TLla Ab in benzyl benzoate after 8 days at 2-8 °C. FIG. 9C shows anti-TLla Ab at 250 mg / mL in benzyl benzoate, the Ab was chemically dehydrated with (5) 30% ethyl lactate / 70% 1 -pentanol and (10) 100% 1 -pentanol. FIG. 9D shows exemplary anti-TLla suspensions after 8 days storage at 2-8 °C, folloyved by inversion 10 times to resuspend, and sitting for 5 min. The Ab dehydrated with (5) 30% ethyl lactate / 70% 1-pentanol remained a homogeneous suspension while the Ab dehydrated with (10) 100% 1-pentanol rapidly fell out of suspension. FIG. 9E shows anti-TLla Ab dehydrated with 30% ethyl lactate / 70% 1- pentanol at 300 mg / mL mAb suspension in triplicate after removal from 2-8 °C (FIG. 9F) and 5 min after inversion resuspension (FIG. 9G). Anti-TLla Ab in PFS at (FIG. 9G - left) 300 mg / mL suspension or (FIG. 9G - right) 290 mg / mL aqueous solution. The PFSs were inverted and a picture was taken when the air bubble (marked in center with a circle) reached the top of the first PFS.

[0022] FIG. 10 is a line graph showing the injection force for anti-TLla dehydrated with 100% pentanol (sample 4 - bottom line) or 30% ethyl lactate / 70% 1-pentanol (sample 3 - top line).

[0023] FIGs. 11-14 are SEM images from Example 2.

[0024] FIG. 15 is a bar graph of the particle size distribution of the particles of Example 2.

[0025] FIG. 16 is a graph of the compression force for the samples in Experiment 2 of Example 5.

[0026] FIG. 17 is a bar graph of the typical field of view of spherical particles detected from anti-TLla Ab dehydrated using 100% 1 -pentanol and sieved through 75 micron mesh of Example 6.

[0027] FIG. 18 is a bar graph of the typical field of view of spherical particles detected from anti-TLla Ab dehydrated using 30% ethyl lactate / 1 -pentanol and sieved through 75 micron mesh of Example 6.

[0028] FIG. 19 are SEM images showing the edge and shape detection results from samples A (left) and B (right) of Example 6. Middle images are enhanced to differentiate the particle shapes and sizes. Bottom images are enhanced to illustrate particle boundaries.

[0029] FIG. 20 is an injection plot for the samples of Example 8 at both concentrations.DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0030] The disclosed processes, dehydrated antibody-containing formulations, suspensions, dosage forms, and compositions may be understood more readily by reference to the following detailed description taken in connection with the accompanying figures, which form a part of this disclosure.

[0031] Throughout this text, where the disclosure describes or claims a feature or embodiment associated with a process for dehydrating an antibody-containing formulation or a process for forming a high concentration antibody suspension, such a feature or embodiment is equally applicable to the dehydrated antibody-containing formulations, dosage forms, suspensions, and compositions. Likewise, where the disclosure describes or claims a feature or embodiment associated with a dehydrated antibody-containing formulation, dosage form, suspension, or composition, such a feature or embodiment is equally applicable to the processes for dehydrating the antibody-containing formulations or the processes for forming the high concentration antibody suspensions.

[0032] Where a range of numerical values is recited or established herein, the range includes the endpoints thereof and all the individual integers and fractions within the range, and also includes each of the narrower ranges therein formed by all the various possible combinations of those endpoints and internal integers and fractions to form subgroups of thelarger group of values within the stated range to the same extent as if each of those narrower ranges was explicitly recited. Where a range of numerical values is stated herein as being greater than a stated value, the range is nevertheless finite and is bounded on its upper end by a value that is operable within the context of the herein disclosure. Where a range of numerical values is stated herein as being less than a stated value, the range is nevertheless bounded on its lower end by a non-zero value. It is not intended that the scope of the processes, dehydrated antibody-containing formulations, suspensions, dosage forms, or compositions be limited to the specific values recited when defining a range. All ranges are inclusive and combinable.

[0033] When values are expressed as approximations, by use of the antecedent “about / ’ it will be understood that the particular value forms another embodiment. Reference to a particular numerical value includes at least that particular value, unless the context clearly dictates otherwise. The term “about” when used in reference to numerical ranges, cutoffs, or specific values is used to indicate that the recited values may vary by up to as much as 10% from the listed value. Thus, the term “about” is used to encompass variations of ± 10% or less from the specified value.

[0034] As used herein, the singular forms “a,” “an,” and “the” include the plural.

[0035] Various terms relating to aspects of the description are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein.

[0036] The term “comprising” is intended to include examples encompassed by the terms “consisting essentially of and “consisting of;” similarly, the term “consisting essentially of’ is intended to include examples encompassed by the term “consisting of.”

[0037] The term “antibody” is meant in a broad sense and includes full length immunoglobulin molecules, antigen-binding fragments thereof, and antibodies conjugated to another molecule, such as antibody -protein fusions and antibody-drug conjugates. Antibody, as used herein, also includes within its scope multispecific (e.g., bispecific, trispecific, etc.) antibodies and multivalent antibodies.

[0038] Immunoglobulins can be assigned to five major classes, namely IgA, IgD, IgE, IgG, and IgM, depending on the heavy chain constant domain amino acid sequence. IgA and IgG are further sub-classified as the isotypes IgAl, IgA2, IgGl, IgG2, IgG3, and IgG4. Antibody light chains of any vertebrate species can be assigned to one of two clearly distincttypes, namely kappa (K) and lambda ( / .). based on the amino acid sequences of their constant domains.

[0039] “Antigen-binding fragment’’ refers to a portion of an immunoglobulin molecule that retains the antigen binding properties of the parental full length antibody (i.e., “antigen-binding fragment thereof’). Exemplary7antigen binding fragments can have: heavychain complementarity determining regions (CDR) 1, 2, and / or 3; light chain CDR 1, 2, and / or 3; a heavy chain variable region (VH); a light chain variable region (VL); and combinations thereof. Antigen binding fragments include: a Fab fragment, a monovalent fragment consisting of the VL, VH, constant light (CL), and constant heavy 1 (CHI) domains; a F(ab)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment consisting of the VH and CHI domains; a Fv fragment consisting of the VL and VH domains of a single arm of an antibody; and a domain antibody (dAb) fragment (Ward et al., Nature 341 :544-546, 1989), which consists of a VH domain or a VL domain. VH and VL domains can be engineered and linked together via a synthetic linker to form various types of single chain antibody designs where the VH / VL domains pair intramolecularly, or intermolecularly in those cases when the VH and VL domains are expressed by separate single chain antibody constructs, to form a monovalent antigen binding site, such as single chain Fv (scFv) or diabody, described in, e.g., Int’l Pub. Nos. W01998 / 44001, WO1988 / 01649, WO1994 / 13804, and W01992 / 01047. These antibody fragments are obtained using techniques well known to those of skill in the art, and the fragments are screened for utility in the same manner as are full length antibodies.

[0040] “Monoclonal antibody” (mAb) refers to a population of antibody molecules of a substantially single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope, or in a case of a bispecific monoclonal antibody, a dual binding specificity7to two distinct epitopes. Monoclonal antibody therefore refers to an antibody population with single amino acid composition in each heavy and each light chain, except for possible well known alterations such as removal of C-terminal lysine from the antibody heavy chain. Monoclonal antibodies may have heterogeneous glycosylation within the antibody population. Monoclonal antibody may be monospecific or multispecific, or monovalent, bivalent or multivalent. A bispecific antibody is included in the term monoclonal antibody.

[0041] The antibody can be a full-length immunoglobulin molecule. Alternatively, the antibody can be an antigen-binding fragment. The antibody can be an antibody-protein fusion. The antibody can be an antibody-drug conjugate.

[0042] In some embodiments, the antibody comprises an anti-IL-5 antibody, anti- IL-23 antibody, anti-PD-1 antibody, an anti-TL la antibody, an anti-CGRP antibody, or an anti-PAR2 antibody. In some embodiments, the antibody comprises an anti-PD-1 antibody. Examples of anti-PD-1 antibodies include, without limitation, pembrolizumab (Keytruda®), nivolumab (Opdivo®), and cemiplimab (Libtayo®). In some embodiments, the antibody comprises an anti-TL la antibody. In some embodiments, the antibody comprises an anti- CGRP antibody. Examples of anti-CGRP antibodies include, without limitation, erenumab (Aimovig®), eptinezumab, galcanezumab (Emgality®), and fremanezumab (Ajovy®). In some embodiments, the antibody comprises an anti-PAR2 antibody. An example of an anti- PAR2 antibody is MEDI0618. In some embodiments, the antibody comprises an anti -IL-5 antibody. Examples of anti-IL-5 antibodies include, without limitation, mepolizumab (Nucala®), reslizumab (Cinqair®), and benralizumab (Fasenra®). In some embodiments, the antibody comprises an anti-IL-23 antibody. Examples of anti-IL-23 antibodies include, without limitation, risankizumab-rzaa (Skyrizi®), guselkumab (Tremfya®), tildrakizumab (Ilumya®). and mirikizumab (Omvoh®).Processes

[0043] The present disclosure provides processes for dehydrating antibodycontaining formulations. Advantageously, the antibodies dehydrated as described herein are of sufficient size to be injected through a syringe. In some embodiments, at least about 60 % w / w of the dehydrated antibody particles are less than 75 microns. In contrast, dehydrated antibodies prepared using 1 -pentanol alone resulted in about 85% w / w of particles that were greater than 75 microns in size, i.e., at least about 85% w / w of the particles cannot be injected through a syringe. The desirable particle size results from the use of the cosolvent that prevents flocculation / aggregation as discussed herein.

[0044] The processes comprise introducing an antibody-containing formulation into a dehydrating solvent mixture comprising 1 -pentanol and a cosolvent that is an ester, wherein the v / v ratio of the 1-pentanol to the cosolvent is about 70 to about 90: about 30 to about 10, to thereby dehydrate the antibody-containing formulation. In some embodiments, the introducing is performed by mixing the antibody-containing formulation and the dehydrating solvent mixture. In some embodiments, the introducing is performed using homogenization, or a porous glass membrane. In some embodiments, the introducing is performed using homogenization. In some embodiments, the introducing is performed using porous glass membranes, such as a porous silica membrane, or such as a Shirasu porous glass (SPG)membrane, optionally including a 2 pm pore syringe adapter. The introducing may be performed at a rate that results in the dehydration of the anti body -containing formulation. In some embodiments, the introducing is performed at a rate that results in flocculation in the dehydrated antibody-containing formulation. For an emulsifying membrane, such as SPG with a 2 pm pore size, the volume per unit time may be scaled linearly according to the surface area of the membrane, expressed in liters per square meter hour (L / (m2*h)), abbreviated as LMH. In some embodiments, the introducing is performed at a rate of about 50 LMH to about 400 LMH. In some embodiments, the introducing is performed at a rate of about 50 LMH, about 100 LMH, about 150 LMH, about 200 LMH. about 250 LMH, about 300 LMH, about 350 LMH, or about 400 LMH. In some embodiments, the introducing is performed at a rate of about 50 to about 350 LMH, about 50 to about 300 LMH, about 50 to about 250 LMH, about 50 to about 200 LMH, about 50 to about 150 LMH, about 50 to about 100 LMH, about 100 to about 400 LMH, about 100 to about 350 LMH, about 100 to about300 LMH, about 100 to about 250 LMH, about 100 to about 200 LMH, about 100 to about150 LMH, about 150 to about 400 LMH, about 150 to about 350 LMH, about 150 to about300 LMH, about 150 to about 250 LMH, about 150 to about 200 LMH, about 200 to about400 LMH, about 200 to about 350 LMH, about 200 to about 300 LMH, about 200 to about250 LMH, about 250 to about 400 LMH, about 250 to about 350 LMH, about 250 to about300 LMH, about 300 to about 400 LMH, about 300 to about 350 LMH, or about 350 to about 400 LMH. In some embodiments, the rate is less than about 375 LMH, less than about 350 LMH, less than about 325 LMH, less than about 300 LMH, less than about 250 LMH, less than 200 LMH, less than about 150 LMH, or less than about 100 LMH. In some embodiments, the rate is about 350 to about 355 LMH. In some embodiments, the introducing is performed at a rate of about 300 to about 400 LMH. In some embodiments, the introducing is performed at a rate of about 350 LMH or less.

[0045] Examples of esters suitable for cosolvents include acetates or lactates. In some embodiments, the cosolvent is an acetate. Suitable acetates include, but are not limited to, ethyl acetate, butyl acetate, heptyl acetate, octyl acetate, or benzyl acetate. In some aspects, the cosolvent is benzyl acetate. In some embodiments, the cosolvent is a lactate. Suitable lactates include, but are not limited to, ethyl lactate. The ethyl lactate can be, for example, ethyl D-lactate, or ethyl D,L-lactate. When not specified, “ethyl lactate” is ethyl-D- lactate. In some aspects, the dehydrating solvent comprises 1 -pentanol and ethyl lactate.

[0046] In some embodiments, the v / v ratio of 1 -pentanol to the cosolvent is about 70 to about 90: about 30 to about 10. In other words, the v / v ratio can be about 70% of 1- pentanol : about 30% of cosolvent, to about 90% 1 -pentanol : about 10% of cosolvent. In some embodiments, the v / v ratio of the 1-pentanol to the cosolvent is about 90: about 10. In some embodiments, the v / v ratio of the 1-pentanol to the cosolvent is about 80: about 20. In some embodiments, the v / v ratio to the 1-pentanol to the cosolvent is about 70: about 30. In some embodiments, the cosolvent is an acetate and the v / v ratio of the 1-pentanol to the acetate is about 80 to about 90: about 20 to about 10. In some embodiments, the v / v ratio of the 1-pentanol to the acetate is about 90: about 10. In some embodiments, the v / v ratio of the 1-pentanol to the acetate is about 80: about 20. In some embodiments, the cosolvent is a lactate and the v / v ratio of the 1-pentanol to the lactate is about 70 to about 90: about 30 to about 10. In some embodiments, the v / v ratio of the 1-pentanol to the lactate is about 90: about 10. In some embodiments, the v / v ratio of the 1-pentanol to the lactate is about 80: about 20. In some embodiments, the v / v ratio to the 1-pentanol to the lactate is about 70: about 30.

[0047] The processes may further comprise separating a liquid phase compnsing the dehydrating solvent from a solid phase comprising the dehydrated antibody-containing formulation. Separating can be performed, for example, using fdtration, decanting, centrifugation, evacuation (e.g, vacuum) among others. In some embodiments, the processes further comprise removing the liquid phase. The processes may include several steps of separating liquid phases from solid phases.

[0048] Once separated, the solid phase may be washed. In some embodiments, the solid phase is washed using a liquid that lacks water. In some embodiments, the solid phase is washed using dehydrated ethanol. In some embodiments, the solid phase is washed using 200 proof ethanol. In some embodiments, the solid phase is washed using dehydrated ethanol containing about 0 v / v%, about 1 v / v%, about 5 v / v%, or about 10 v / v% of water.

[0049] The processes may also include one or more optional dry ing steps. In some embodiments, the solid phase is dried under reduced pressure. Such techniques include, without limitation, the use of vacuums at varying temperatures and pressures.

[0050] The processes may further include one or more optional cooling and / or heating steps. In some embodiments, the solutions, solids, or a combination thereof are cooled by reduced temperatures. For example, the cooling may be performed to reduce the temperature of a solution and / or solid to temperatures above 0°C. such as about 2 to about 8°C. Cooling may be performed using ice baths, refrigerators, cooling rooms or the like.

[0051] The term ‘‘dehydrated” as used herein with reference to the “dehydrated antibody-containing formulation” refers to the lack of water in the solid dehydrated antibodycontaining formulation. In some embodiments, the dehydrated antibody-containing formulations contain less than about 5 w / w% of water, based on the weight of the formulation. In some embodiments, the dehydrated antibody-containing formulations contain less than about 4 w / w%, less than about 3 w / w%, less than about 2 w / w%, less than about 1 w / w%. less than about 0.5 w / w%, or less than about 0. 1 w / w% of water, based on the weight of the formulation. In some embodiments, the dehydrated antibody-containing formulations contain about 0. 1 w / w% to about 5 w / w% of water, based on the weight of the formulation. In further embodiments, the dehydrated antibody-containing formulations contain about 0. 1 w / w% to about 4 w / w%. about 0. 1 w / w% to about 3 w / w%, about 0. 1 w / w% to about 2 w / w%, about 0.1 w / w% to about 1 w / w%, about 0.1 w / w% to about 0.5 w / w%, about 0.5 w / w% to about 5 w / w%, about 0.5 w / w% to about 4 w / w%, about 0.5 w / w% to about 3 w / w%, about 0.5 w / w% to about 2 w / w%, about 0.5 w / w% to about 1 w / w%, about 1 w / w% to about 5 w / w%, about 1 w / w% to about 4 w / w%, about 1 w / w% to about 3 w / w%, about 1 w / w% to about 2 w / w%. about 2 w / w% to about 5 w / w%, about 2 w / w% to about 4 w / w%, about 2 w / w% to about 3 w / w%, about 3 w / w% to about 5 w / w%, about 3 w / w% to about 4 w / w%, or about 4 w / w% to about 5 w / w% of water, based on the weight of the formulation.

[0052] The dehydrated antibody-containing formulation can comprise antibodycontaining microbeads. The antibody-containing microbeads can be small, uniform beads that remain in suspension for long lengths of time and can be injected through a syringe, including subcutaneously administered. The term “uniform,” with reference to the beads, refers to a consistent bead structure throughout the antibody-containing formulation. “Uniform” includes, for example, a lack of fractures, occlusions, and flocculations. “Uniform” also includes, for example, a similar particle size. Such fractures, occlusions, and flocculations may be viewed using techniques in the art including, without limitation, scanning electron microscopy (SEM).

[0053] In some embodiments, the antibody-containing microbeads have an average diameter of less than about 30 pm. In some embodiments, the antibody-containing microbeads have an average diameter of less than about 25 pm, less than about 20 pm, less than about 15 pm, less than about 10 pm, or less than about 5 pm. In some embodiments, the antibody-containing microbeads have an average diameter of about 1 pm to about 25 pm. In some embodiments, the antibody-containing microbeads have an average diameter of about 1 pm to about 20 pm, about 1 pm to about 15 pm, about 1 pm to about 10 pm, about 1 pm toabout 5 un, about 5 un to about 25 pun, about 5 pun to about 20 pun, about 5 pun to about 15 pun, about 5 pun to about 10 pun. about 10 pun to about 25 pun, about 10 pun to about 20 pun, about 10 pun to about 15 pun, about 15 pun to about 25 pun, about 15 pun to about 20 pun, or about 20 pun to about 25 pm. In some embodiments, the antibody-containing microbeads have an average diameter of about 1 pun to about 20 pun. In some embodiments, the antibody-containing microbeads have an average diameter of about 29 pun, about 28 pun, about 27 pun. about 26 pun, about 25 pun, about 24 pun, about 23 pun. about 22 pun. about 21 pun, about 20 pun, about 19 pun, about 18 pun, about 17 pun, about 16 pun, about 15 pun, about 14 pun, about 13 pun, about 12 pun, about 11 pun, about 10 pun, about 9 pun, about 8 pun, about 7 pun, about 6 pun, about 5 pun, about 4 pun. about 3 pun, about 2 pun, or about 1 pirn.

[0054] Prior to dehydration, the antibody -containing formulations may also contain other components. In some embodiments, the antibody-containing formulations contain sucrose. In some embodiments, the anti-IL-5 antibody-containing formulations, anti-PD-1 antibody-containing formulations, or anti-CGRP antibody-containing formulations contain sucrose.

[0055] Also disclosed herein are processes for forming a high concentration antibody suspension, the processes comprising: dehydrating an antibody -containing formulation according to any of the herein disclosed processes to form a dehydrated antibody-containing formulation and resuspending the dehydrated antibody-containing formulation in a vehicle to form the high concentration antibody suspension.

[0056] The dehydrated antibody -containing formulation used in the process for forming a high concentration antibody suspension can comprise antibody-containing microbeads. The antibody-containing microbeads can have an average diameter of less than about 30 pm. such as about 1 pm to about 20 pm. or such as about 3 pm to about 7 pm. In some embodiments, the antibody-containing microbeads have an average diameter of less than about 30 pm, less than about 28 pm, less than about 26 pm, less than about 24 pm, less than about 22 pm, less than about 20 pm, less than about 18 pm, less than about 16 pm, less than about 14 pm, less than about 12 pm, less than about 10 pm, less than about 8 pm, less than about 6 pm. less than about 4 pm, less than about 2 pm, or less than about 1 pm. In some embodiments, at least about 90% of the antibody-containing microbeads have an average diameter of less than about 30 pm.

[0057] Suitable vehicles include those disclosed herein, such as benzy l benzoate, medium chain triglyceride, ethyl lactate, ethyl oleate, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane.isopropyl myristate, triglyceride of caprylic and capric acid, or combinations thereof. In some embodiments, the vehicle is benzyl benzoate.

[0058] The high concentration antibody suspensions produced by the disclosed methods can contain at least about 100 mg / mL of the antibody. In some embodiments, the suspension contains about 150 mg / mL to about 400 mg / mL of the antibody. In some embodiments, the suspension contains at least about 100 mg / mL, at least about 150 mg / mL, at least about 200 mg / mL, at least about 250 mg / mL, at least about 300 mg / mL, at least about 350 mg / mL, or at least about 400 mg / mL of the antibody. In some embodiments, the suspension contains about 100 mg / mL to about 400 mg / mL, about 100 mg / mL to about 375 mg / mL, about 100 mg / mL to about 350 mg / mL, about 100 mg / mL to about 325 mg / mL, about 100 mg / mL to about 300 mg / mL, about 100 mg / mL to about 275 mg / mL, about 100 mg / mL to about 250 mg / mL, about 100 mg / mL to about 200 mg / mL, about 100 mg / mL to about 140 mg / mL, about 150 mg / mL to about 400 mg / mL, about 150 mg / mL to about 375 mg / mL, about 150 mg / mL to about 350 mg / mL, about 150 mg / mL to about 300 mg / mL, about 150 mg / mL to about 275 mg / mL, about 150 mg / mL to about 250 mg / mL, about 150 mg / mL to about 200 mg / mL, about 175 mg / mL to about 400 mg / mL. about 175 mg / mL to about 375 mg / mL, about 175 mg / mL to about 350 mg / mL, about 175 mg / mL to about 300 mg / mL, about 175 mg / mL to about 275 mg / mL, about 175 mg / mL to about 250 mg / mL, about 175 mg / mL to about 200 mg / mL, about 200 mg / mL to about 400 mg / mL, about 200 mg / mL to about 375 mg / mL, about 200 mg / mL to about 350 mg / mL. about 200 mg / mL to about 300 mg / mL, about 200 mg / mL to about 275 mg / mL, about 200 mg / mL to about 250 mg / mL, about 225 mg / mL to about 400 mg / mL, about 225 mg / mL to about 375 mg / mL, about 225 mg / mL to about 350 mg / mL, about 225 mg / mL to about 300 mg / mL, about 225 mg / mL to about 275 mg / mL, about 225 mg / mL to about 250 mg / mL. about 250 mg / mL to about 400 mg / mL, about 250 mg / mL to about 375 mg / mL, about 250 mg / mL to about 350 mg / mL, about 250 mg / mL to about 325 mg / mL, about 250 mg / mL to about 300 mg / mL, about 250 mg / mL to about 275 mg / mL, about 275 mg / mL to about 400 mg / mL, about 275 mg / mL to about 375 mg / mL, about 275 mg / mL to about 350 mg / mL. about 275 mg / mL to about 325 mg / mL. about 275 mg / mL to about 300 mg / mL, about 300 mg / mL to about 400 mg / mL, about 300 mg / mL to about 375 mg / mL, about 300 mg / mL to about 350 mg / mL, about 300 mg / mL to about 325 mg / mL, about 325 mg / mL to about 400 mg / mL, about 325 mg / mL to about 375 mg / mL, about 325 mg / mL to about 350 mg / mL, about 350 mg / mL to about 400 mg / mL, about 350 mg / mL to about 375 mg / mL, or about 375 mg / mL to about 400 mg / mL of the antibody.

[0059] The high concentration antibody suspensions generated by the disclosed processes can be delivered to a subject in low volumes (such as equal to or less than about 2 mL), and are therefore suitable for subcutaneous delivery. Antibody therapeutics that are typically administered by intravenous routes can therefore be concentrated using the disclosed processes and administered subcutaneously.Dehydrated Antibody-Containing Formulations and Suspensions Comprising The Same

[0060] Disclosed herein are dehydrated antibody-containing formulations prepared according to any of the herein disclosed processes. In some embodiments, the dehydrated antibody-containing formulations comprise an anti-PD-1 antibody. In some embodiments, the dehydrated antibody-containing formulations comprise an anti-TLla antibody. In some embodiments, the dehydrated antibody-containing formulations comprise an anti-CGRP antibody. In some embodiments, the dehydrated antibody-containing formulations comprise an anti-PAR2 antibody.

[0061] The dehydrated antibody-containing formulations comprise about 40% to about 95% w / w of antibody, based on the weight of the formulation. In some embodiments, the dehydrated antibody-containing formulations comprise about 40% w / w, about 50% w / w, about 60% w / w, about 70% w / w, about 80% w / w, about 90% w / w, or about 95% w / w of antibody, based on the weight of the formulation. In some embodiments, the dehydrated antibody-containing formulations comprise about 40% w / w to about 80% w / w, about 40% w / w to about 60% w / w, about 50% w / w to about 95% w / w, about 50% w / w to about 80% w / w, about 60% w / w to about 95% w / w, or about 60% w / w to about 80% w / w of antibody, based on the weight of the formulation.

[0062] The dehydrated antibody-containing formulations can be resuspended in a vehicle to generate a suspension. Thus, disclosed herein are suspensions prepared according to any of the herein disclosed methods. In some embodiments, the suspensions comprise antibody-containing microbeads. In some embodiments, the vehicle is benzyl benzoate, benzyl alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of caprylic and capric acid, or combinations thereof. In some embodiments, the vehicle is benzy l benzoate. In some embodiments, the vehicle is benzyl alcohol. In some embodiments, the vehicle is a medium chain triglyceride. In some embodiments, the vehicleis ethyl lactate. In some embodiments, the vehicle is ethyl oleate. In some embodiments, the vehicle is polyethylene glycol 200. In some embodiments, the vehicle is propylene glycol. In some embodiments, the vehicle is a perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane. In some embodiments, the vehicle is isopropyl myristate. In some embodiments, the vehicle is a triglyceride of caprylic and capric acid.

[0063] The suspension can comprise less than about 0.5 area%. based on the area% of the antibody in the antibody-containing formulation prior to the dehydration, of antibody multimers, such as dimerized antibody. The term “antibody multimer” as used herein refers to a compound that includes two or more antibodies that are chemically bound together. The term “dimerized antibody” as used herein refers to a compound that includes two antibodies that are chemically bound together. In some embodiments, the suspension comprises less than about 0.4 area%, less than about 0.3 area%, less than about 0.2 area%, or less than about 0. 1 area%, based on the area% of the antibody in the antibody-containing formulation prior to the dehydration, of dimerized antibody. In other embodiments, the suspension comprises about 0.01 area% to about 0.5 area%, based on the area% of the antibody in the antibodycontaining formulation prior to the dehydration, of dimerized antibody. In further embodiments, the suspension comprises about 0.01 area% to about 0.4 area%, about 0.01 area% to about 0.3 area%, about 0.01 area% to about 0.2 area%, about 0.01 area% to about 0. 1 area%, about 0.01 area% to about 0.05 area%. about 0.05 area% to about 0.5 area%, about 0.05 area% to about 0.4 area%, about 0.05 area% to about 0.3 area%, about 0.05 area% to about 0.2 area%, about 0.05 area% to about 0.1 area%, about 0.1 area% to about 0.5 area%, about 0.1 area% to about 0.4 area%, about 0.1 area% to about 0.3 area%, about 0.1 area% to about 0.2 area%, about 0.2 area% to about 0.5 area%, about 0.2 area% to about 0.4 area%, about 0.2 area% to about 0.3 area%, about 0.3 area% to about 0.5 area%, about 0.3 area% to about 0.4 area%, or about 0.4 area% to about 0.5 area%, based on the area% of the antibody in the antibody-containing formulation prior to the dehydration, of dimerized antibody.

[0064] The suspension can have about 10% or less separation for up to about 2 hours, as compared to a control suspension that was formed by dehydrating the antibodycontaining formulation using 100% 1 -pentanol. The term “separation” as used herein refers to a chemical state where the solid and liquid materials physically separate from each other. In some embodiments, the suspension has about 9%, about 8%. about 7%, about 6%. about 5%, about 4%, about 3%, about 2%, or about 1% or less separation for up to about 2 hours, ascompared to a control suspension that was formed by dehydrating the antibody-containing formulation using 100% 1 -pentanol. In some embodiments, the suspension has about 1% to about 10% less separation for up to about 2 hours, as compared to a control suspension that was formed by dehydrating the antibody-containing formulation using 100% 1 -pentanol. In some embodiments, the suspension has about 1% to about 8%, about 1% to about 6%, about 1% to about 4%, about 1% to about 2%, about 2% to about 10%, about 2% to about 8%, about 2% to about 6%. about 2% to about 4%. about 4% to about 10%. about 4% to about 8%, about 4% to about 6%, about 6% to about 10%, about 6% to about 8%, or about 8% to about 10% less separation for up to about 2 hours, as compared to a control suspension that was formed by dehydrating the antibody-containing formulation using 100% 1-pentanol.

[0065] The suspensions can comprise 1-pentanol, a cosolvent, and an antibodycontaining formulation. In some embodiments, the suspension comprises an antibodycontaining formulation, and a dehydrating solvent comprising 1-pentanol and a cosolvent that is an ester, such as an acetate or a lactate. The v / v ratio of the 1-pentanol to the cosolvent can be as described herein, such as about 70 to about 90: about 30 to about 10. Thus, disclosed herein are suspensions comprises a dehydrating solvent comprising 1-pentanol and a cosolvent that is an ester, such as an acetate or a lactate, wherein the v / v ratio of the 1- pentanol to the cosolvent is about 70 to about 90: about 30 to about 10, and an antibodycontaining formulation. In some aspects, the antibody-containing formulation is dehydrated. In some aspects, the antibody-containing formulation is partially dehydrated. In some aspects, the antibody-containing formulation has not yet been dehydrated. In some embodiments, the dehydrating solvent comprises 1-pentanol and ethyl lactate.

[0066] The v / v ratio of the 1-pentanol to the cosolvent can be about 70: about 30. or about 80: about 20, or about 90: about 10.

[0067] The suspension may contain microbeads that contain the antibodies. In some embodiments, at least a portion of the antibody-containing formulation is in a form of antibody-containing microbeads. In some aspects, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%. at least about 60%, at least about 70%. at least about 80%. at least about 90%, at least about 95%, at least about 99%, or about 100% of the antibody-containing formulation is in a form of antibody-containing microbeads.

[0068] The suspensions can contain about 150 mg / mL to about 400 mg / mL of the antibody. In some embodiments, the suspension contains at least about 150 mg / mL, at least about 200 mg / mL, at least about 250 mg / mL, at least about 300 mg / mL, at least about 350 mg / mL, or at least about 400 mg / mL of the antibody. In some embodiments, the suspensioncontains about 150 mg / rnL to about 400 mg / mL, about 150 mg / rnL to about 375 mg / rnL. about 150 mg / mL to about 350 mg / mL, about 150 mg / mL to about 300 mg / mL, about 150 mg / mL to about 275 mg / mL, about 150 mg / mL to about 250 mg / mL, about 150 mg / mL to about 200 mg / mL, about 175 mg / mL to about 400 mg / mL, about 175 mg / mL to about 375 mg / rnL, about 175 mg / mL to about 350 mg / mL, about 175 mg / mL to about 300 mg / mL, about 175 mg / mL to about 275 mg / mL, about 175 mg / mL to about 250 mg / mL, about 175 mg / mL to about 200 mg / mL, about 200 mg / mL to about 400 mg / mL. about 200 mg / mL to about 375 mg / mL, about 200 mg / mL to about 350 mg / mL, about 200 mg / mL to about 300 mg / rnL, about 200 mg / mL to about 275 mg / mL, about 200 mg / mL to about 250 mg / mL, about 225 mg / mL to about 400 mg / mL, about 225 mg / mL to about 375 mg / mL, about 225 mg / mL to about 350 mg / mL, about 225 mg / mL to about 300 mg / mL. about 225 mg / mL to about 275 mg / mL, about 225 mg / mL to about 250 mg / mL, about 250 mg / mL to about 400 mg / mL, about 250 mg / mL to about 375 mg / mL, about 250 mg / mL to about 350 mg / mL, about 250 mg / mL to about 325 mg / mL, about 250 mg / mL to about 300 mg / mL, about 250 mg / mL to about 275 mg / mL, about 275 mg / mL to about 400 mg / mL. about 275 mg / mL to about 375 mg / mL. about 275 mg / mL to about 350 mg / mL, about 275 mg / mL to about 325 mg / mL, about 275 mg / mL to about 300 mg / mL, about 300 mg / mL to about 400 mg / mL, about 300 mg / mL to about 375 mg / mL, about 300 mg / mL to about 350 mg / mL, about 300 mg / mL to about 325 mg / mL, about 325 mg / mL to about 400 mg / mL. about 325 mg / mL to about 375 mg / mL, about 325 mg / mL to about 350 mg / mL, about 350 mg / mL to about 400 mg / mL, about 350 mg / mL to about 375 mg / mL, or about 375 mg / mL to about 400 mg / mL of the antibody.Compositions

[0069] Provided herein are compositions comprising a vehicle and a plurality of antibody-containing microbeads. The plurality of antibody-containing microbeads comprise at least about 150 mg / mL of an antibody. In some embodiments, the plurality of antibodycontaining microbeads comprise about 150 mg / mL to about 400 mg / mL of the antibody. In some embodiments, the plurality of antibody-containing microbeads contain at least about 150 mg / mL, at least about 200 mg / mL, at least about 250 mg / mL, at least about 300 mg / mL, at least about 350 mg / mL, or at least about 400 mg / mL of the antibody. In some embodiments, the plurality of antibody-containing microbeads contains about 150 mg / mL to about 400 mg / mL, about 150 mg / mL to about 375 mg / mL, about 150 mg / mL to about 350 mg / mL, about 150 mg / mL to about 300 mg / mL, about 150 mg / mL to about 275 mg / mL,about 150 mg / mL to about 250 mg / mL, about 150 mg / mL to about 200 mg / mL, about 175 mg / mL to about 400 mg / mL, about 175 mg / mL to about 375 mg / mL. about 175 mg / mL to about 350 mg / mL, about 175 mg / mL to about 300 mg / mL, about 175 mg / mL to about 275 mg / mL, about 175 mg / mL to about 250 mg / mL, about 175 mg / mL to about 200 mg / mL, about 200 mg / mL to about 400 mg / mL, about 200 mg / mL to about 375 mg / mL, about 200 mg / mL to about 350 mg / mL, about 200 mg / mL to about 300 mg / mL. about 200 mg / mL to about 275 mg / mL. about 200 mg / mL to about 250 mg / mL, about 225 mg / mL to about 400 mg / mL, about 225 mg / mL to about 375 mg / mL, about 225 mg / mL to about 350 mg / mL, about 225 mg / mL to about 300 mg / mL, about 225 mg / mL to about 275 mg / mL, about 225 mg / mL to about 250 mg / mL, about 250 mg / mL to about 400 mg / mL. about 250 mg / mL to about 375 mg / mL, about 250 mg / mL to about 350 mg / mL, about 250 mg / mL to about 325 mg / mL, about 250 mg / mL to about 300 mg / mL, about 250 mg / mL to about 275 mg / mL, about 275 mg / mL to about 400 mg / mL, about 275 mg / mL to about 375 mg / mL, about 275 mg / mL to about 350 mg / mL, about 275 mg / mL to about 325 mg / mL, about 275 mg / mL to about 300 mg / mL, about 300 mg / mL to about 400 mg / mL, about 300 mg / mL to about 375 mg / mL, about 300 mg / mL to about 350 mg / mL. about 300 mg / mL to about 325 mg / mL. about 325 mg / mL to about 400 mg / mL, about 325 mg / mL to about 375 mg / mL, about 325 mg / mL to about 350 mg / mL, about 350 mg / mL to about 400 mg / mL, about 350 mg / mL to about 375 mg / mL, or about 375 mg / mL to about 400 mg / mL of the antibody.

[0070] The aforementioned compositions have one or more of the following characteristics as compared to a control composition comprising antibody-containing microbeads formed using 100% 1 -pentanol: an injection force of less than about 45 N when suspended in benzyl benzoate and injected through a 27 G manual prefilled syringe (PFS); a reduced density compared to the control composition as exemplified in FIG. 9; a reduced percentage of flocculation as compared to the control composition as exemplified in FIGs. 3 and 4; or about 10% or less separation for up to about 2 hours as compared to the control composition as exemplified in FIG. 9..

[0071] The compositions may also have one or more of the following characteristics as compared to a control composition comprising antibody-containing microbeads formed using 100% 1 -pentanol:an injection force of about 2 to about 20 N when sieved with a 75 pm sieve, suspended in benzyl benzoate, and injected through a 27 G manual prefilled syringe (PFS); about 60% total weight of the material being < 75 microns (usable material) as compared to about 85% total weight of the material being > 75 microns (unusable material) when formed using 100% 1 -pentanol; a reduced settling of particles (about 80 % weight of particles formed using 100% 1-pentanol are >75 micron and settle in about 2 minutes or less as compared to about 40 % weight of particles formed using ethyl lactate and 1 -pentanol which are >75 micron; a reduced settling (higher sedimentation rate) compared to a control composition as exemplified in FIG. 9; a greater particle uniformity as compared to particles formed using 100% 1- pentanol (75-micron sieved powder showed an increase of greater than 50% in particles with fractures and flocculations when dehydrated with 100% 1- pentanol alone; a greater powder uniformity as compared to particles formed using 100% 1- pentanol (94% of particles in a 75-micron sieved pow der were within a 30 pm range as compared to 87% of particles in a 75-micron sieved powder formed using 100% 1-pentanol alone); smaller 75-micron sieved particle sizes as compared to particles formed using 100% 1-pentanol (particle sizes of 15 um as compared to particle sizes of 25 um when formed using 100% 1-pentanol); or a reduced percentage of flocculation as compared to the control composition as exemplified in FIGs. 3 and 4.

[0072] In some embodiments, the compositions have an injection force of about 2 to about 20 N when sieved with a 75 pm sieve, suspended in benzy l benzoate, and injected through a 27 G manual prefilled syringe (PFS), as compared to a control composition comprising antibody-containing microbeads formed using 100% 1-pentanol. In some embodiments, the compositions have an injection force of about 8N when sieved w ith a 75 pm sieve, suspended in benzyd benzoate, at a concentration of about 100 pg / mL and injected through a 27 G manual prefilled syringe (PFS), as compared to a control composition comprising antibody-containing microbeads formed using 100% 1-pentanol. In some embodiments, the compositions have an injection force of about 14N when sieved with a 75pm sieve, suspended in benzyl benzoate, at a concentration of about 400 pg / mL and injected through a 27 G manual prefilled syringe (PFS), as compared to a control composition comprising antibody-containing microbeads formed using 100% 1 -pentanol. In some embodiments, about 60% total weight of the material of the compositions are < 75 microns (usable material) as compared to about 85% total weight of the material being > 75 microns (unusable material) when formed using 100% 1 -pentanol. In some embodiments, the compositions have a reduced settling of particles (about 80 % weight of particles formed using 100% 1-pentanol are >75 micron and settle in about 2 minutes or less as compared to about 40 % weight of particles formed using ethyl lactate and 1-pentanol which are >75 micron. In some embodiments, the compositions have a reduced settling (higher sedimentation rate) compared to a control composition as exemplified in FIG. 9. In some embodiments, the compositions have a greater particle uniformity as compared to particles formed using 100% 1-pentanol (75-micron sieved powder showed an increase of greater than 50% in particles with fractures and flocculations when dehydrated with 100% 1-pentanol alone. In some embodiments, the compositions have a greater powder uniformity as compared to particles formed using 100% 1-pentanol (94% of particles in a 75-micron sieved powder were within a 30 pm range as compared to 87% of particles in a 75-micron sieved powder formed using 100% 1-pentanol alone). In some embodiments, the compositions have smaller 75-micron sieved particle sizes as compared to particles formed using 100% 1- pentanol (particle sizes of 15 um as compared to particle sizes of 25 um when formed using 100% 1 -pentanol). In some embodiments, the compositions have a reduced percentage of flocculation as compared to the control composition as exemplified in FIGs. 3 and 4. In some embodiments, the compositions have about 10% or less separation for up to about 2 hours as compared to the control composition as exemplified in FIG. 9.

[0073] The antibody-containing microbeads can have an average diameter of less than about 30 pm, such as about 1 to about 20 pm.

[0074] Suitable vehicles include, for example, benzyl benzoate, medium chain triglyceride, ethyl lactate, ethyl oleate, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of caprylic and capric acid, or combinations thereof. In some embodiments, the vehicle is benzy l benzoate.Dosage Forms

[0075] The disclosure also provides dosage forms comprising any of the suspensions described herein. In some embodiments, the dosage form is a subcutaneous dosage form. For example, the subcutaneous dosage form can be formulated for administration by injection or infusion. The dosage form may include, e.g., hypodermic needle / syringe, injector pen, pump / canula, among others, as determined by the attending physician.

[0076] The dosage forms can comprise antibody-containing microbeads prepared as described herein. In some embodiments, the antibody-containing microbeads are prepared by dehydrating an antibody-containing formulation. The dosage forms can comprise suspensions that comprise the antibody -containing microbeads prepared as described herein. In some embodiments, the concentration of the antibody in the antibody-containing microbead suspension is at least about 150 mg / mL, such as up to about 400 mg / mL.

[0077] Such dosage forms may have advantageously low viscosity. The ability to provide large doses of antibody in a small volume, with low viscosity, permits administration, e.g. subcutaneously.Methods

[0078] The cosolvent assists in protecting against or preventing aggregation that may be caused by 1 -pentanol. Moreover, the cosolvent provided a dehydrated Ab that, upon reconstitution and injection, resulted in at least 3 times the amount of Ab expelled through the injection needle.

[0079] Thus, disclosed herein are methods of reducing antibody aggregation during dehydration with 1 -pentanol, the methods comprising introducing an antibody-containing formulation into a dehydrating solvent mixture comprising 1 -pentanol and ethyl lactate, wherein the v / v ratio of the 1 -pentanol to the ethyl lactate is about 70 to about 90: about 30 to about 10, to thereby reduce the aggregation of the antibody.Aspects I

[0080] Aspect IL A process for dehydrating an antibody-containing formulation, the process comprising: introducing the antibody-containing formulation into a dehydrating solvent mixture comprising 1-pentanol and a cosolvent that is an ester, wherein the v / v ratio of the 1-pentanol to the cosolvent is about 70 to about 90: about 30 to about 10, to thereby dehydrate the antibody-containing formulation.

[0081] Aspect 12: The process of Aspect II, wherein the ester is an acetate or lactate.

[0082] Aspect 13: The process of Aspect II or 12, wherein the ester is an acetate.

[0083] Aspect 14: The process of Aspect 13, wherein the acetate is ethyl acetate, butyl acetate, heptyl acetate, octyl acetate, or benzyl acetate, such as benzyl acetate.

[0084] Aspect 15: The process of Aspect II or 12, wherein the ester is a lactate.

[0085] Aspect 16: The process of Aspect 15. wherein the lactate is ethyl lactate, such as ethyl D-lactate, ethyl L-lactate, or ethyl D,L-lactate.

[0086] Aspect 17 : The process of any one of the preceding Aspects I, wherein the ratio of the 1-pentanol to the ester is about 90: about 10, about 80: about 20, or about 70: about 30.

[0087] Aspect 18: The process of any one of the preceding Aspects I, wherein the dehydrating solvent comprises 1-pentanol and ethyl lactate.

[0088] Aspect 19: The process of any one of the preceding Aspects I, wherein the dehydrated antibody-containing formulation comprises antibody-containing microbeads.

[0089] Aspect 110: The process of Aspect 19, wherein the antibody-containing microbeads have an average diameter of less than about 30 pm, such as about 1 pm to about 20 pm.

[0090] Aspect Il l : The process of any one of the preceding Aspects I, wherein the introducing is performed using homogenization, a Shirasu porous glass membrane, or microfluidics.

[0091] Aspect 112: The process of any one of the preceding Aspects I, wherein the introducing is performed at a rate of less than 50 pL / min.

[0092] Aspect 113: The process of any one of the preceding Aspects I, further comprising separating a liquid phase comprising the dehydrating solvent from a solid phase comprising the dehydrated antibody -containing formulation.

[0093] Aspect 114: The process of Aspect 113, further comprising removing the liquid phase.

[0094] Aspect 115: The process of Aspect 114. further comprising washing the solid phase.

[0095] Aspect 116: The process of Aspect 115, wherein the washing is performed using ethanol, such as 200 proof ethanol.

[0096] Aspect 117: The process of any one of the preceding Aspects I, further comprising resuspending the dehydrated antibody-containing formulation in a vehicle to generate a suspension comprising antibody-containing microbeads.

[0097] Aspect 118: The process of Aspect 117, wherein the vehicle is benzy l benzoate, benzy l alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200. propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of caprylic and capric acid, or combinations thereof.

[0098] Aspect 119: The process of Aspect 118, wherein the vehicle is benzy l benzoate.

[0099] Aspect 120: The process of any one of Aspects 117-119, wherein the suspension comprises less than about 0.5 area%, based on the area% of the antibody in the antibody-containing formulation prior to the dehydration, of dimerized antibody.

[0100] Aspect 121: The process of any one of Aspects 117-120. wherein the suspension has about 10% or less separation for up to about 2 hours, as compared to a control suspension that was formed by dehydrating the antibody-containing formulation using 100% 1 -pentanol.

[0101] Aspect 122: A dehydrated antibody-containing formulation prepared according to the process of any one of Aspects 11-116.

[0102] Aspect 123: The dehydrated antibody-containing formulation of Aspect 122, wherein the antibody comprises an anti-PD-1 antibody, an anti-TLl a antibody, an anti-CGRP antibody, or an anti-PAR2 antibody.

[0103] Aspect 124: A suspension prepared according to the process of any one of Aspects 117-121.

[0104] Aspect 125: A dosage form comprising the suspension of Aspect 124.

[0105] Aspect 126: The dosage form of Aspect 125, which is a subcutaneous dosage form.

[0106] Aspect 127: The dosage form of Aspect 125 or 126, wherein the dosage form comprises antibody-containing microbeads.

[0107] Aspect 128: The dosage form of Aspect 127, wherein the concentration of the antibody in the antibody-containing microbead suspension is at least about 150 mg / mL, such as up to about 400 mg / mL.

[0108] Aspect 129: A suspension comprising:a dehydrating solvent comprising 1 -pentanol and a cosolvent that is an ester, wherein the v / v ratio of the 1-pentanol to the cosolvent is about 70 to about 90: about 30 to about 10, and an antibody-containing formulation.

[0109] Aspect 130: The suspension of Aspect 129, wherein the v / v ratio of the 1- pentanol to the cosolvent is about 70: about 30, or about 80: about 20, or about 90: about 10.

[0110] Aspect 131 : The suspension of Aspect 129 or 130. wherein the ester is an acetate or lactate.

[0111] Aspect I32:The suspension of any one of Aspects 129-131 , wherein the dehydrating solvent comprises 1-pentanol and ethyl lactate.

[0112] Aspect 133: The suspension of any one of Aspects 129-132. wherein at least a portion of the antibody-containing formulation is in a form of antibody-containing microbeads.

[0113] Aspect 134: A composition comprising: a vehicle and a plurality of antibody-containing microbeads, wherein the plurality of antibody -containing microbeads comprise at least about 150 mg / mL of an antibody and wherein the composition has one or more of the following characteristics as compared to a control composition comprising antibodycontaining microbeads formed using 100% 1-pentanol: an injection force of less than about 45 N when suspended in benzyl benzoate and injected through a 27 G manual prefilled syringe (PFS); a reduced density' compared to the control composition as exemplified in FIG. 9; a reduced percentage of flocculation as compared to the control composition as exemplified in FIGs. 3 and 4; or about 10% or less separation for up to about 2 hours as compared to the control composition as exemplified in FIG. 9.

[0114] Aspect 135: The composition of Aspect 134, wherein the antibodycontaining microbeads have an average diameter of less than about 30 pm, such as about 1 to about 20 pm.

[0115] Aspect 136: The composition of Aspect 134 or 135, wherein the plurality of antibody-containing microbeads comprise at least about 150 mg / mL, at least about 200 mg / mL, at least about 250 mg / mL, at least about 300 mg / mL, or at least about 350 mg / mL, or at least about 400 mg / mL of the antibody.

[0116] Aspect 137: The composition of any one of Aspects 134-136, wherein the vehicle is benzyl benzoate, benzyl alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of capry lic and capric acid, or combinations thereof.

[0117] Aspect 38: A process for forming a high concentration antibody suspension, the process comprising: dehydrating an antibody-containing formulation according to the process of any one of Aspects 11-116 to form a dehydrated antibody-containing formulation; and resuspending the dehydrated antibody-containing formulation in a vehicle to form the high concentration antibody suspension.

[0118] Aspect 139: The process of Aspect 138, wherein the dehydrated antibodycontaining formulation comprises antibody-containing microbeads.

[0119] Aspect 140: The process of Aspect 139, wherein the antibody-containing microbeads have an average diameter of less than about 30 pm. such as about 1 to about 20 pm.

[0120] Aspect 141: The process of any one of Aspects 138-140, wherein the vehicle is benzyl benzoate, benzyl alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of capry lic and capric acid, or combinations thereof.Aspects II

[0121] Aspect III: A process for dehydrating an antibody-containing formulation, the process comprising: introducing the anti body -containing formulation into a dehy drating solvent mixture comprising 1-pentanol and a cosolvent that is an ester, wherein the v / v ratio of the 1-pentanol to the cosolvent is about 70 to about 90: about 30 to about 10, to thereby dehydrate the antibody-containing formulation.

[0122] Aspect II-2: The process of Aspect III, wherein the ester is an acetate or lactate.

[0123] Aspect 113: The process of Aspect III or 112, wherein the ester is an acetate.

[0124] Aspect 114: The process of Aspect 113, wherein the acetate is ethyl acetate, butyl acetate, heptyl acetate, octyl acetate, or benzyl acetate, such as benzyl acetate.

[0125] Aspect 115: The process of Aspect III or 112, wherein the ester is a lactate.

[0126] Aspect 116: The process of Aspect 115, wherein the lactate is ethyl lactate, such as ethyl D-lactate, ethyl L-lactate, or ethyl D,L-lactate.

[0127] Aspect 117: The process of any one of the preceding Aspects II, wherein the ratio of the I-pentanol to the ester is about 90: about 10, about 80: about 20, or about 70: about 30.

[0128] Aspect 118: The process of any one of the preceding Aspects II, wherein the dehydrating solvent comprises 1-pentanol and ethyl lactate.

[0129] Aspect 119: The process of any one of the preceding Aspects II, wherein the dehydrated antibody-containing formulation comprises antibody-containing microbeads.

[0130] Aspect III 0: The process of Aspect 119, wherein the antibody-containing microbeads have an average diameter of less than about 30 pm, such as about 1 pm to about 20 pm.

[0131] Aspect III 1: The process of any one of the preceding Aspects II. wherein the introducing is performed using homogenization, a Shirasu porous glass membrane, or microfluidics.

[0132] Aspect 1112: The process of any one of the preceding Aspects II, wherein the introducing is performed at a rate of less than 50 pL / min.

[0133] Aspect III 3: The process of any one of the preceding Aspects II, wherein the antibody-containing formulation further comprises sucrose.

[0134] Aspect 1114: The process of any one of the preceding Aspects II, further comprising separating a liquid phase comprising the dehydrating solvent from a solid phase comprising the dehydrated antibody -containing formulation.

[0135] Aspect 1115: The process of Aspect 1114, further comprising removing the liquid phase.

[0136] Aspect 1116: The process of Aspect 1114, further comprising washing the solid phase.

[0137] Aspect 1117: The process of Aspect 1116, wherein the washing is performed using ethanol, such as 200 proof ethanol.

[0138] Aspect 1118: The process of any one of the preceding Aspects II, further comprising resuspending the dehydrated antibody-containing formulation in a vehicle to generate a suspension comprising antibody-containing microbeads.

[0139] Aspect 1119: The process of Aspect 1118, wherein the vehicle is benzyl benzoate, benzyl alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of caprylic and capric acid, or combinations thereof.

[0140] Aspect 1120: The process of Aspect III 8, wherein the vehicle is benzyl benzoate.

[0141] Aspect 1121 : The process of any one of Aspects III 8-II20, wherein the suspension comprises less than about 0.5 area%, based on the area% of the antibody in the antibody-containing formulation prior to the dehydration, of dimerized antibody.

[0142] Aspect 1122: A dehydrated antibody-containing formulation prepared according to the process of any one of Aspects III -1121.

[0143] Aspect 1123: The dehydrated antibody-containing formulation of Aspect 1122, wherein the antibody comprises an anti-IL-5 antibody, anti-IL-23 antibody, anti-PD-1 antibody, an anti-TLla antibody, an anti-CGRP antibody, or an anti-PAR2 antibody.

[0144] Aspect 1124: A suspension prepared according to the process of any one of Aspects III 8-II21.

[0145] Aspect 1125: A dosage form comprising the suspension of Aspect 1124.

[0146] Aspect 1126: The dosage form of Aspect 1125. which is a subcutaneous dosage form.

[0147] Aspect 1127: The dosage form of Aspect 1125 or 1126, wherein the dosage form comprises antibody-containing microbeads.

[0148] Aspect 1128: The dosage form of Aspect 1127. wherein the concentration of the antibody in the antibody-containing microbead suspension is at least about 150 mg / mL, such as up to about 400 mg / mL.

[0149] Aspect 1129: A suspension comprising: a dehydrating solvent comprising 1 -pentanol and a cosolvent that is an ester, wherein the v / v ratio of the 1 -pentanol to the cosolvent is about 70 to about 90: about 30 to about 10. and an antibody-containing formulation.

[0150] Aspect 1130: The suspension of Aspect 1129, wherein the v / v ratio of the 1- pentanol to the cosolvent is about 70: about 30, or about 80: about 20, or about 90: about 10.

[0151] Aspect 1131 : The suspension of Aspect 1129 or 1130, wherein the ester is an acetate or lactate.

[0152] Aspect 1132: The suspension of any one of Aspects II29-II31 , wherein the dehydrating solvent comprises 1 -pentanol and ethyl lactate.

[0153] Aspect 1133: The suspension of any one of Aspects II29-II32, wherein at least a portion of the antibody-containing formulation is in a form of antibody-containing microbeads.

[0154] Aspect 1134: A composition comprising: a vehicle and a plurality of antibody-containing microbeads, wherein the composition has one or more of the following characteristics as compared to a control composition comprising antibody-containing microbeads formed using 100% 1- pentanol: an injection force of about 2 to about 20 N when sieved with a 75 pm sieve, suspended in benzyl benzoate, and injected through a 27 G manual prefilled syringe (PFS); about 60% total weight of the material being < 75 microns (usable material) as compared to about 85% total weight of the material being > 75 microns (unusable material) when formed using 100% 1 -pentanol; a reduced settling of particles (about 80 % weight of particles formed using 100% 1 -pentanol are >75 micron and settle in about 2 minutes or less as compared to about 40 % weight of particles formed using ethyl lactate and 1 -pentanol which are >75 micron; a reduced settling (higher sedimentation rate) compared to a control composition as exemplified in FIG. 9; a greater particle uniformity' as compared to particles formed using 100% 1- pentanol (75-micron sieved powder showed an increase of greater than 50% in particles with fractures and flocculations when dehydrated with 100% 1- pentanol alone; a greater powder uniformity7as compared to particles formed using 100% 1- pentanol (94% of particles in a 75-micron sieved powder were within a 30 pm range as compared to 87% of particles in a 75-micron sieved powder formed using 100% 1 -pentanol alone); smaller 75-micron sieved particle sizes as compared to particles formed using 100% 1 -pentanol (particle sizes of 15 um as compared to particle sizes of 25 um when formed using 100% 1 -pentanol); ora reduced percentage of flocculation as compared to the control composition as exemplified in FIGs. 3 and 4.

[0155] Aspect 1135: The composition of Aspect 1134, wherein the antibodycontaining microbeads have an average diameter of less than about 30 pm, such as about 1 to about 20 pm.

[0156] Aspect 1136: The composition of Aspect 1134 or 1135, wherein the vehicle is benzyl benzoate, benzyl alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of capry lic and capric acid, or combinations thereof.

[0157] Aspect 37: A process for forming a high concentration antibody suspension, the process comprising: dehydrating an antibody-containing formulation according to the process of any one of Aspects III -II 17 to form a dehydrated antibody-containing formulation; and resuspending the dehydrated antibody-containing formulation in a vehicle to form the high concentration antibody suspension.

[0158] Aspect 1138: The process of Aspect 1137, wherein the dehydrated antibodycontaining formulation comprises antibody-containing microbeads.

[0159] Aspect 1139: The process of Aspect 1138, wherein the antibody-containing microbeads have an average diameter of less than about 30 pm, such as about 1 to about 20 pm.

[0160] Aspect 1140: The process of any one of Aspects II37-II39, wherein the vehicle is benzyl benzoate, benzyl alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of capry lic and capric acid, or combinations thereof.Examples

[0161] The following examples are provided to further describe some of the embodiments disclosed herein. The examples are intended to illustrate, not to limit, the disclosed embodiments.Example 1A. Materials and Procedures

[0162] The antibodies listed in Table 1 were used in these examples.

[0163] SPG syringe adapter was purchased through Fisher Scientific from ASONE (Japanese).

[0164] All mAbs were used as received without changing the buffer composition as shown in Table 1. mAbs were diluted to the operating concentration with MiliQ or WFI water, unless otherwise indicated.

[0165] Buffer for anti-TLla Ab, anti-CGRP Ab, and anti-PAR2 Ab were prepared. See, Table 1.

[0166] (i) Vortexing Procedure for Chemical Dehydration

[0167] Procedure A: 20 pL of anti-PD-1 Ab (25 mg / mL, allowed to come to room temperature for 30 min before starting) in buffer (10 mM histidine, 7.0% w / v sucrose, 0.02% w / v polysorbate 80, pH 5.5) was added to 1 mL of the organic solvents in Table 2, and then vortexed for 30 seconds at 3000 rpm. The samples were then centrifuged at 10.000 RPM for 60 seconds. The supernatant was removed, a hole was punctured in the top of the tube and closed, and the samples were placed under high-vacuum for 90 min in a lyophilizer jar with aluminum foil wrapping. The samples were then reconstituted in 20 pL of water and 90 pL of formulation buffer (buffer filtered through 0.2 pm syringe filter before use) then vortexed for 30 seconds at 3000 rpm. The samples were then centrifuged at 10,000 RPM for 120 seconds, transferred to a PCR tube and the sample concentration was determined using A280. A 20 pL of stock anti-PD-1 Ab diluted with 90 pL of formulation buffer then centrifuged at 10,000 RPM for 120 seconds, was prepared and used as a control. All solutions remained clear except sample 25 which was dehydrated with ethanol / ethyl lactate solution combination.

[0168] Procedure B: 10 pL of anti-PD-1 Ab (25 mg / mL, allowed to come to room temperature for 30 min before starting) in buffer (10 mm histidine, 7.0% w / v sucrose, 0.02% w / v polysorbate 80, pH 5.5) was added to 0.5 mL of the organic solvents in Table 2, and then vortexed for 30 seconds at 3000 rpm. The dehydrated antibody was allowed the settle and the organic solvent was removed with a pipette tip attached to a vacuum flask. The pellet was broken up and resuspended in benzy l benzoate. The tip of a 200 pL pipette was cut off, 10 pL of sample was placed on a slide with a glass top then images were captured at 40X magnification.

[0169] Procedure C: 50 pL of the diluted anti-CGRP Ab (75 mg / mL) was added to 1 mL of the organic solvent on Table 2 in the bottom of a 1.5 mL tube and vortexed at 3000 rpm for 30 seconds. The precipitate was pelleted by centrifuging the samples at 10000 rpm for 5 min and the solvent was removed with a vacuum flask. The pellet was washed by adding 400 pL of 200 proof ethanol, vortexed for 10 seconds at 3000 rpm, pelleted by centrifuge, and the ethanol was removed by vacuum flask. Four holes were punctured in the top of the Eppendorf tubes and they were placed on high vacuum for 48 hrs.

[0170] (ii) Reconstitutions

[0171] Chemically dehydrated powder was reconstituted in buffer by pipetting. The aqueous mAh solution was then filtered through a 0.22 pm syringe filter and assayed with the assays listed in table below (Table 3).

[0172] (ii) SPG Membrane Procedure for Chemical Dehydration

[0173] Procedure A - Initial assessment: Anti-PD-1 Ab (25 mg / mL) was taken up into a 1 rnL BD plastic syringe, the syringe was then inverted and the SPG membrane was placed on the end of the syringe. The air was fully expelled. The syringe pump was placed on the following settings: infuse only mode, BD plastic 1 mL syringe, 0.1 mL / min, target volume is 250 pL. The syringe was attached to the syringe pump, inverted, and placed in a 10 rnL blue polycarbonate bottle containing 8 mL of 1 -pentanol with a stir bar on a stir plate set to 400 rpm. After bead formation the suspension was allowed to mix for an additional 3 min to allow full dehydration, the beads were allowed to settle and the organic solvent was decanted off. The containers were then placed under high vacuum for 24 h then placed at 2-8 °C over the weekend.

[0174] Procedure B - Process Scale-Up: A 3.5 mL of mAb was diluted to 7 mL with WFI water. The mAh solution was taken up into a 10 mL syringe, inverted, and the air was removed. A syringe extension tubing was connected to the end of the syringe and the air was removed replaced with the diluted mAb liquid. The SPG membrane adapter was then attached to the end of the extension tubing and the air was gently pressed out of the system. The exterior of the SPG membrane was dabbed with a ChemWipe to remove residual liquid on membrane face. The syringe was then loaded into a syringe pump and the SPG membrane was lowered into the center of a 125 mL Erlenmeyer flask containing 110 mL of 30% ethyl lactate / 1 -pentanol with a stir-bar set to 300 rpm. The pump was set to 25 pL / min and the pump was started. Upon completion, the solution was poured into a 50 mL conical tube and centrifuged at 1500 rpm for 5 min. The organic liquid was decanted and more of the productwas poured in, centrifuged, and decanted. This was repeated until all product was collected in the 50 mL conical tube. 20 mL of 200 proof ethanol was added to the tube, the tube capped, and the product was resuspended and washed by gently inverting the tube back and forth. The sample was centrifuged at 1500 rpm and the ethanol was removed using a vacuum flask with a pipette tip. The sample was washed two additional times (totaling 3 wash cycles). After removal of ethanol, holes were placed in the cap of the tube using a 24 g needle and the cap was placed on the 50 mL conical tube. A ChemWipe was placed over the cap and secured with a rubber band. The sample was placed in a lyophilization jar and placed on high vacuum for ~48 hrs.B. Results and discussion

[0175] (i) Results with Single Solvents

[0176] Work was performed using anti-PD-1 Ab in the histidine-based buffer to identify dehydrating organic liquids that would not impact chemical dehydration or mAb stability'. 20 pL of anti-PD-1 Ab was homogenized, by vortexing (procedure A), into various organic liquids with observations recorded (Table 4). 1-pentanol and 1-octanol led to the dehydration of the micro-droplets into microbeads, although there were visibly more microbeads present for the 1-pentanol dehydration compared to 1-octanol. Ethanol and 2- propanol led to a turbid solution, suggesting mAb instability. Benzyl alcohol led to a clear solution, while toluene did not absorb water and remained phase separated. All of the alcohols, except for toluene, solvated water but only 1-pentanol and 1-octanol led to microbead formation. Acetates were also examined and ethyl acetate dehydrated the mAb into a single large pellet. Larger acetates, butyl-octyl and benzy l acetates, led to a biphasic mixture, with the aqueous on the bottom and organic layer on top.

[0177] Interestingly, the ability to form microbeads does not clearly follow solvation. Fully miscible solvents form turbid solutions. Microbeads were formed in 1- pentanol (22g / L) and 1 -octanol (almost insoluble), but not in ethyl acetate, diethyl ether, benzyl alcohol, or butyl acetate nor other solvents with very low water solubility.

[0178] (ii) Results with Solvent mixtures

[0179] After establishing that chemical dehydration was compatible with 1 - pentanol and 1 -octanol, dehydration properties were analyzed by mixing alcohols with acetate. Initial tests were run using 50 / 50 mixtures of alcohol / acetate. Results showed that ethyl and butyl acetates in 1 -pentanol led to a single dehydrated pellet while heptyl and octyl acetates led to a biphasic solution. Both ethyl lactate and benzyl acetate led to microbead formation. 2-propanol and ethanol mostly lead to turbid solutions, suggesting low mAb stability'. Observations are summarized in Table 5 below.

[0180] Further studies utilizing 10% ethyl acetate, butyl acetate, heptyl acetate, octyl acetate, benzyl benzoate, benzyl alcohol, or ethyl lactate in 90% 1 -pentanol all led to microbead formation.

[0181] (iii) Analysis of Dehydrated Antibodies

[0182] Microscopy and A280 (after reconstitution) were used to analyze the dehydrated mAb powders prepared as described above. Dehydrated mAb powder from conditions found to form microbeads or possible microbeads were reconstituted in buffer. Analysis by A280 revealed >90% yield for all conditions except 1 -octanol which exhibited 41% yield, relative to the non-dehydrated control (Table 6). This was consistent with the observation that the pellet from octanol dehydration was much smaller than the other solvent and solvent combinations (results not shown). Residual organic solvent can inflate concentrations above a 100 % of 5 mg / mL. Full reconstitution is later demonstrated by SEC (Table 8) as well by total area under the curve compared to the control.

[0183] Microscopy was performed on all conditions that lead to microbead formation (Figure 1). 1-pentanol led to microbeads -15-25 pm in diameter. Ethyl and butyl acetate in 1-pentanol led to irregular microbeads or amorphous material. Heptyl acetate in 1- pentanol led to uniform microbeads -5-13 pm in diameter while octyl acetate in 1-pentanol led to uniform microbeads -10-15 pm in diameter. However, flocculation was observed for all of these conditions. The addition of benzyl, ethyl, and butyl acetates all led to irregular microbeads.

[0184] (iv) Solvent Ratio Analysis

[0185] The results showed that at 20% and greater of heptyl, octyl, and benzyl acetates, and benzy l benzoate, anti-PD-1 Ab fused into a single pellet. By increasing the cosolvents to 40%, the organic liquid became biphasic. Interestingly, 10% ethyl lactate (mixture of isomers) in 1 -pentanol led to small uniform microbeads (FIGs. 2A-2D). As the ethyl lactate content increased, the microbeads became more uniform and smaller in size.

[0186] At 50% ethyl lactate / 1 -pentanol, the solution became turbid. SEC performed on this sample showed a higher population of aggregates. Further, the dehydrated mAb was challenging to reconstitute (consistent with insoluble aggregation) and the sample lacked clearly defined microbeads.

[0187] The dehydrated anti-PD-1 Ab reconstituted in buffer was then analyzed by SEC (Table 7).

[0188] Anti-PD-1 Ab exhibited >99.5% monomer populations (by SEC) after the homogenization and various dehydrating organic solutions, suggesting it is fully stable to the tested conditions. Anti-PD-1 Ab dehydrated with 10% ethyl lactate exhibited slightly- elevated H3 populations (0.06%). Increasing the ethyl lactate content to up to 40% lead to a 9% decrease in the monomer population (Table 7).

[0189] SEC was then performed on the same samples. All samples showed greater than 90% recovery, except for a 50:50 mixture of n-pentanol / benzyl acetate. See, Table 8.

[0190] (v) SPG Membrane

[0191] The homogenization was then analyzed. In particular, aqueous mAbs (in the buffers noted in Table 1) were homogenized through a SPG membrane into the dehydrating solution. A single use SPG membrane syringe adapter, syringe pump, and syringe with aqueous mAb was utilized for homogenization. First, the flow rate of the syringe pump loaded with anti-PD-1 Ab at 25 mg / mL was used with procedure A. At a flow rate of 100 pL / min, microbeads were observed to coalesce around the outer side of the SPG membrane, leading to flocculated beads. The flow rate was decreased to 50 pL / min and less flocculation was observed leading to a fine homogenous powder. Decreasing the flow rate further to 10 pL / min lead to a single pellet and few microbeads.

[0192] Homogenization was then performed using anti-TLla Ab. Anti-TLla Ab was diluted in half to 75 mg / mL with water for injection (WFI) and the flow rate of the syringe pump was examined. Using 100 pL / min, large droplets coalesced on the outer face of the SPG membrane indicating the flow rate was too high for small droplets to fall off the membrane face. The flow rate was increased to 150, 200, and 300 pL / min where flocculation became increasingly severe. With 25 mg / mL anti-PD-1 Ab, a flow rate of 25-50 pL / min was preferable for microbead formation without flocculation. With 75 mg / mL anti-TLla Ab, all conditions led to flocculation and large conglomerates (FIGs. 3A-3C). However, dehydration solvents utilizing 30% ethyl lactate all led to uniform anti-TLla Ab powder (FIGs. 3D, 3F,and 3G). The anti-TLla Ab powder was suspended in benzyl benzoate and analyzed by microscopy confirming the severe flocculation caused by 1 -pentanol alone (FIG. 4A). All samples dehydrated with 30% ethyl lactate solutions exhibited insignificant flocculation (FIGs. 4C, 4E, and 4G). Beyond flocculation, the dehydration solvent exhibited a large impact on microbead morphology. Anti-TLla Ab dehydrated in 1-pentanol alone appeared clear with diameters -20-40 pm, while anti-TLla Ab dehydrated in 30% ethyl lactate was opaque with diameters of -2-4 pm (major) and -10-20 pm (minor) (FIGs. 4B and 4D). Given the SPG pore size is 1 pm, this data indicates the ethyl lactate helps to maintain small microbead size and prevents coalescing of the micro-droplets.

[0193] To confirm the stability of anti-TLla Ab, dehydrated samples under various conditions were reconstituted in buffer and analyzed by SEC (Table 10). The data indicated that anti-TLla Ab was slightly sensitive to ethyl lactate, yet maintained an acceptable 98.6% monomer populations. Dehydrated anti-TLla Ab, from 30% ethyl lactate / 70% 1-pentanol, was washed three times with 200 proof ethanol then dried on vacuum. The sample was analyzed by SEC indicating that dehydration of the mAb was stable to 200 proof ethanol washes.

[0194] (vi) Process Scale-up

[0195] The scalability and biophysical analysis of mAbs anti-TLla Ab (150 mg / mL), anti-CGRP Ab (150 mg / mL), and anti-PAR2 Ab (100 mg / mL) was conducted. mAbs were diluted to a half of their starting concentration and 7 mL of each was loaded into a 10 mL syringe for homogenization into 110 mL of 30% ethyl lactate / 1-pentanol. All dehydrated mAbs resuspended homogeneously in the ethanol wash, indicating no flocculation. mAb powders were placed under high vacuum at room temperature for 48 hr. To determine the efficiency of the ethanol wash and high vacuum, anti-TLla Ab and anti- PAR2 Ab were dissolved at 50 mg / mL, then dissolved to 1 mg / mL. and analyzed by GCMSusing standard curves. Reconstituted anti-TLla Ab solution at 50 mg / mL contained 1.679 mg / rnL of ethanol, 0.481 mg / mL of 1-pentanol, and 0.574 mg / mL of ethyl lactate. Reconstituted anti-PAR2 Ab solution at 50 mg / mL contained 1.581 mg / rnL ethanol, 0.927 mg / rnL 1-pentanol, and 1.038 mg / mL ethyl lactate.

[0196] The process scale-up discussed above produced >500 mg of the mAbs with comparable stoichiometry between the mAbs and formulates to the unmodified aqueous mAb stocks (the theoretical mAb %w / w to all molecules present is comparable to the calculated mAb %w / w of the powder; z.e., only water was removed during the process) (Table 11). The dehydrated mAb was analyzed by Karl Fisher, which indicated the presence of 2-3% water. The powder was very' fine and exhibited no flocculation. By microscopy, all three mAbs exhibited similar particle size with the majority of particles being ~2-4 pm and minor particles being -10-15 pm, again with no observed flocculation. As a control, anti-TLla Ab was chemically dehydrated under the same process using 100% 1-pentanol. Surprisingly, and in contrast to all previous studies, the powder resuspended homogenously during the ethanol wash and no flocculation was observed by microscopy (FIG. 5D). Using a SPG membrane, flow rate of 25 pL / min, and 8 mL of dehydration liquid, severe flocculation was observed. In contrast, maintaining the same flow rate with 110 mL of dehydration liquid led to no flocculation. This finding is in contrast to literature reports using a lab scale SPG membrane and pump, where faster dehydration rates led to less flocculation. The microbead size was much larger at approximately 20-30 pm for anti-LKla Ab dehydrated with 1-pentanol alone.

[0197] The dehydrated mAbs were reconstituted in buffer and analyzed for potency and stability. Using an ELISA assay, dehydrated anti-TLla Ab had a reported mean relative potency of 91% indicating comparable potency to the control sample. Using a cell based assay, dehydrated anti-CGRP Ab had a reported mean relative potency of 99%, while anti-PAR2 Ab had a reported mean relative potency of 102%. All mAbs maintained full potency after dehydration and reconstitution. Utilizing circular dichroism spectroscopy,overlaying both the control and dehydrated samples revealed identical secondary structures (FIGs. 6A-6C). Local maximum and minimum wavelengths of the dehydrated and reconstructed samples were consistent with the control samples. The data was further analyzed by secondary7structure prediction software confirming no changes occurred to the secondary structure during the course of dehydration I reconstitution process. The samples were then examined by DSC to determine any loss in thermal stability due to the process. Both anti-TLla Ab and anti-CGRP Ab exhibited identical DSC results between the control and dehydrated samples, consistent with CD and no change in structure due to the dehydration process (FIGs. 7A-7C). Anti-PAR2 Ab exhibited comparable Tml and Tm2 to the control sample. However, the overall peak shape and Tm3 was not consistent with the control. Although the DSC data suggested a change in tertiary structure of potentially CH3 domain, the CD data along with Tm1 (potentially CH2 domain) and Tm2 (potentially Fc domain) from DSC was consistent, with no change in tertiary (DSC) and secondary (CD) structure. Data from CGE and icIEF indicated that the mAbs are fully intact, exhibiting comparable results between all dehydrated mAbs and controls. By using DLS, anti-PAR2 Ab exhibited identical hydrodynamic radius to the control while anti-TLla Ab exhibited a small increase of 0.9 nm, suggesting both molecules are stable to aggregation (Table 17). Interestingly, anti-CGRP Ab exhibited a large increase in size from 5.61 nm to 13.32 nm consistent with potential aggregation as there was a multimodal distribution of the light scattering species. All samples exhibited an increase in %PD due to insoluble formulate particles from the dehydration procedure. SEC showed that anti-TLl a Ab and anti-PAR2 Ab exhibited a small decrease in the monomer population to 97.5% and 96.8%, respectively - this is consistent with what was observed using 30% ethyl lactate / 1 -pentanol with anti-PD-1 Ab. Consistent with the DLS results, anti-CGRP Ab exhibited a large decrease in monomer population from 96.6 to 75.5% along with an increase in HMW species from 3. 1 to 24.2%, confirming ethyl lactate induced aggregation.

[0198] (vii) Buffer Dilutions

[0199] Dilution of the mAbs using buffer, instead of water was then analyzed.When anti-CGRP Ab was diluted with buffer (16 mM histidine, 6.6 % w / v sucrose, 0. 136 mg / mL disodium EDTA dihydrate, 0.02 % PS80, pH 5.5), its SEC elution profile was restored from 84.5% monomer population (with water) to 96. 1% by SEC along with a decrease in hydrodynamic radius by DLS from 9.23 nm (with water) to 6.43 nm (FIG. 8, Table 17). Samples diluted with buffer all exhibited comparable monomer populations of 96- 97%. Every dehydration condition where anti-CGRP Ab was diluted with buffer out performed water dilutions significantly.

[0200] As shown above, benzyl benzoate was found to be more appropriate for suspension studies.

[0201] (viii) PFS Suspension and Injectability7

[0202] Anti-CGRP Ab, anti-PAR2 Ab, and anti -TL la Ab (dehydrated with (a) 30% ethyl lactate / 70% 1 -pentanol or (b) 100% 1 -pentanol) powders were suspended at 250 mg / mL of mAb in benzyl benzoate (FIG. 9A). The mAbs were loaded into prefilled syringes (PFSs), capped, and stored at 2-8°C. After 8 days, the suspension settled comparably for all 3 mAbs (FIG. 9B). The PFSs containing anti-TLla Ab dehydrated with 100% 1 -pentanol appeared more densely packed than the PFS containing anti-TLla Ab dehydrated with 30% ethyl lactate / 70% 1 -pentanol, since they have the same weight of material (FIG. 9C). The PFS were inverted ten times to resuspend the mAbs then allowed to sit for 5 min. After 5 min, all mAbs dehydrated with 30% ethyl lactate / 70% 1 -pentanol remained a stable homogenous suspension while anti-TLla Ab samples dehydrated with 100% 1 -pentanol rapidly settled at the bottom of the syringe (FIG. 9D).

[0203] Anti-TLla Ab was examined at 300 mg / mL suspension in PFS in triplicate for 8 days after storage at 2-8°C (FIG. 9E). The samples were resuspended by inverting the PFS and, after 5 min, the mAbs remained a homogenous suspension (FIG. 9F). The 300mg / mL of mAb suspensions remained homogenous for more than 2 h after resuspension, demonstrating the stability of the suspension for the test period.

[0204] PFSs containing anti-TLla Ab as a 300 mg / mL of mAb suspension or a 290 mg / mL of mAb aqueous solution were further examined. Both PFSs were inverted to compare the viscosity of the two mAbs based on displacement of the head space air bubble (FIG. 9G). While the air in the suspension rapidly floated to the top of the PFS, the aqueous formulation took more than twice as long to do so. Based on this visual inspection, the higher concentration suspension at 300 mg / mL mAb had a lower viscosity than the aqueous mAb at 290 mg / mL.

[0205] The break loose (BLF) and extrusion force of the PFSs was then analyzed using aqueous and chemically dehydrated and suspended mAbs. Aqueously dissolved anti- TLla Ab at 250 mg / mL had an average BLF force of 3.40 N and extrusion force of 22.75 N while the 290 mg / mL had an average BLF force of 3.82 N and extrusion force of 49.43 N. Although the concentration increased by 14%, the extrusion force doubled. Benzy l benzoate (in the absence of mAb) in PFSs exhibited an average BLF of 3.25 N and extrusion force of 6.89 N. while ethyl lactate (in the absence of mAb) in PFSs exhibited an average BLF of 3.49 N and extrusion force of 2.65 N.

[0206] The sample dehydrated with 100% pentanol did not exhibit any stable injection force and was consistent with extrusion of the vehicle liquid predominantly while the powder dehydrated with the 30% ethyl lactate mixture exhibited a stable injection force of about 20 N. The term ‘'stable injection” force can be shown when the slope does not change for a given period of time. See, FIG. 10.

[0207] Examining the plots from 4-6 mm suggests the extrusion force of anti-TLla Ab dehydrated with 30% ethyl lactate is ~20 N.

[0208] mAbs also were dehydrated and homogenized with a disposable SPG syringe adapter. 3.5 mL of mAb was diluted to 75 mg / mL with WFI water. The mAb solution was taken up into a 10 mL syringe, inverted, and the air was removed. A syringe extension tubing was connected to the end of the syringe and the air was removed replaced with the diluted liquid mAb. The SPG membrane adapter was then attached to the end of the extension tubing and the air was gently pressed out of the system. The syringe was then loaded into a syringe pump and the SPG membrane was lowered into the center of a 125 mL Erlenmeyer flask containing 110 mL of 30% ethyl lactate / 1 -pentanol with a stir-bar set to 300 rpm. The pump was set to 25 pL / min and the pump was started. Upon completion the solution was poured into a 50 mL conical tube and centrifuged at 1500 rpm for 5 min. Theorganic liquid was decanted and more of the product was poured in, centrifuged, and decanted. This was repeated until all product was collected in the 50 mL conical tube. 20 mL of 200 proof ethanol was added to the tube, capped, and the product was resuspended and washed by gently inverting the tube back and forth. The sample was centrifuged at 1500 rpm and the ethanol was removed using a vacuum flask with a pipette tip. The sample was washed two additional times (totaling 3 wash cycles). After removal of ethanol, holes were placed in the cap of the tube using a 24 g needle and the cap was placed on the 50 mL conical tube. The sample was placed in a lyophilization jar and placed on high vacuum for ~48 hrs.C. Conclusion

[0209] It was found that adding 10-30% ethyl lactate to the 1 -pentanol dehydration solution had several advantages: 1) reduced powder flocculation, 2) smaller particles, 3) reduced microbead density, and 4) a stable homogenous suspension in benzyl benzoate even at 300 mg / mL of mAb. All of these attributes led to a better mAb formulation for S.C. administration as an alternative to the I.V. administration.

[0210] Chemical dehydration is a robust process and was successfully performed on anti-PD-1 Ab, anti-CGRP Ab, anti-PAR2 Ab, and anti-TLla Ab. In contrast to previously reported uses of SPG membranes, it was found that slow flow rates relative to a large organic liquid volume reduces or eliminates flocculation.

[0211] This data indicates chemical dehydration is a viable process. The process has several advantages to traditional mAb formulation development: 1) the mAb formulation is directly dehydrated, stored dry, and administered as a suspension without developing a long term aqueous stable formulation, 2) storing and delivering the mAb dry eliminates certain degradation pathways, 3) allows high concentration delivery independent of the mAb’s viscosity or the need to develop a high concentration liquid formulation, 4) concentration ranges of at least 150 mg / mL are attainable as opposed to aqueous formulations, and 5) eliminates the need for process development to handle high viscosity aqueous mAb formulations.Example 2: Particle Size Analysis

[0212] The sample of anti-IL-5 Ab was a dry' powder and was dispersed across a carbon adhesive tab then coated with a few nm of gold for conductivity.

[0213] Images were taken using a thermal field emission SEM (Leo 1530 TFE). The magnification was chosen to include many particles in each image whilst keepingenough free space between them to measure individual particles. The particle size distribution was measured using a semi -automated analysis algorithm. 1036 particles were measured to determine the average particle size and statistical uncertainty.

[0214] Representative SEM images are shown in FIGs. 11-14, along with FIG. 12 that show s the outlines of the measured particles. Clusters that could not be separated and any particles at the edges of the image were excluded. The overall morphology was very circular, with a relatively broad size distribution and occasional large particles showing dimples.

[0215] The particle size distribution shows the expected log-normal distribution. The average particle size w as 12.4 ± 0.3 pm. See, FIG. 15.Example 3: Chemical Dehydration Stability Studies

[0216] Anti PD-1 Ab. anti-IL-23 Ab. and anti IL-5 Ab were dehydrated as described herein. The samples were first formulated as follows:(i) Anti PD-1 Ab 100 mg / mL, 25 mM acetate, 7.5% trehalose, 0.03% P188(ii) Anti -IL-23 Ab, 152 mg / mL, 10 mM acetate, 7% trehalose, pH 5.8(hi) Anti IL-5 Ab 95 mg / mL, 23 mM acetate, 300 mM sucrose, 0.3 mg / mL PS80, pH5.5

[0217] Each sample was then diluted using 100% water or 7% sucrose / water and dehydrated using 100% 1 -pentanol or ethyl lactate / 1 -pentanol mixtures having the amounts of ethyl lactate shown in Tables 18-20. The stability of each sample was analyzed using size exclusion chromatography (SEC) measuring % of high molecular weight (HMW), % monomer, and % of low molecular weight (LMW). See, Tables 18-20.

[0218] The results in Table 18 show decreasing monomer stability with increasing ethyl lactate %. Dilution with sucrose decreased monomer stability to ethyl lactate. Overall, anti-PD-1 Ab is very stable to the chemical dehydration described herein.

[0219] The results in Table 19 show that decreasing monomer stability with increasing ethyl lactate %. Further, dilution with sucrose decreases monomer stability to ethyl lactate. Overall, anti-IL-23 Ab is very stable to chemical dehydration.

[0220] The results in Table 20 show that increasing then decreasing monomer stability with increasing ethyl lactate %. Ethyl lactate at low % protects anti-IL-5 Ab from chemical dehydration. Moreover, dilution with sucrose increased monomer stability to ethyl lactate. Overall, anti -IL-5 Ab is stable to 1 -pentanol with 20-30 % ethyl lactate. Unexpectedly, ethyl lactate enhanced BOTH mAb stability and powder properties.Example 4: Scale-Up of Anti-IL-5 Ab Dehydration

[0221] Anti-IL-5 Ab (95 mg / mL) was diluted to 72 mg / mL with 7% sucrose then dehydrated as described herein using 25% ethyl L-lactate / 1-pentanol. The dehydration was performed in BSC. The flask, stir bar, and sieve were autoclaved before use. The syringe and extension tubing were sterile and for single use. The emulsification adapter was cleaned with 70 % IPA before dehydration. The dehydrated powder was analyzed as described inTable 21.

[0222] A280 results showed that the mAb content in powder equal to 42 % w / w (indicating 29.76 mg of powder per rat). NMR results showed that no acetate buffer was present and that sucrose content was 41.5 % w / w. Karl Fisher analysis showed 3.02 % w / w water. GC showed residual organic liquid, z.e., ethanol content was 1.36 % w / w. ethyl lactate content was 5.27 % w / w, and 1-pentanol content was 3.09 % w / w. Scanning electron microscopy showed an average particle size of 12.4 pm, with no particles >76 pm. See, FIG. 14.

[0223] The dehydrated anti-IL-5 Ab was reconstituted in and analyzed using dynamic light scattering (DLS), full spectrum fluorescence (FSF), SEC, non-reduced (NR)- CGE, reduced (R)-CGE, and icIEF. DLS showed a DH of 15.7 nm, where the control (z.e., the same anti-IL-5 before dilution and dehydration) was 14.0 nm. See, FIG. 15. FSF showeda Tml of 56.9°C, where the control was 57.5°C. SEC showed 97.2% monomer, where the control was 98.9%. NR-CGE showed 97.7% purity, where the control was 97.9%. R-CGE showed 99.3% purity, wherein the control was 99.2%. icIEF showed 61.7% for the main peak, where the control had a 67.2% for the main peak.Example 5: PFS Experiments for Delivery of Anti-TLla Ab

[0224] (i) Experiment 1

[0225] (a) Sample Preparation

[0226] Dehydrated anti -TL la Ab was prepared as described herein using 100% 1- pentanol (2 samples) or 30% ethyl lactate / 1 -pentanol (6 samples). Each sample was washed three times using ethanol. Samples were then transferred to a funnel connected to a vacuum flask with a filter membrane between. After ethanol removal, the funnel was removed and the filter membrane containing the powder was transferred to a 50 mL conical tube and placed on vacuum. This optimized product recover}'.

[0227] GC of residual alcohols in the dehydrated anti-TLla Ab sample in 20 mM acetate, 7 % sucrose. pH 5.5 showed 0.8% w / w ethanol. 3.2% w / w 1-pentanol, and 5.5% w / w ethyl lactate.

[0228] The washed powders were then loaded onto a 75-micron sieve over weighing paper, thereby separating the sample into sizes less than and greater than 75 microns. The weights of each particle size sample was then measured. See, Table 22.

[0229] The results illustrate that the use 30% ethyl lactate / I -pentanol tripled the yield of product having a particle size of less than 75 microns.

[0230] (b) Injection Testing

[0231] The 30% ethyl lactate / 1 -pentanol anti-TLla Ab powders from part (i) were weighed into 1.5 mL Eppendorf tubes and the volume of mAh powder calculated using density. Benzyl benzoate was then added to each tube to a volume of 500 pL. Each sample was briefly vortexed at 2300 rpm until all powder appeared wet. Samples were taken up in a 1 mL pipette and added to an empty 27G PFS, capped, and dispensed from the PFS via injection. The break loose and extrusion forces were then determined. See, Table 23.

[0232] These data show that the average extrusion force was low at the high concentrations utilized.

[0233] (ii) Experiment 2

[0234] A 1 ml Luer Lock ISO Type 1 glass syringe was utilized to reduce opportunity for needle clogging. The goal was to reduce initial force profile and maintain consistent initiating and glide forces.

[0235] Anti-TLIA Ab (25 mg) that was dehydrated as described herein was reconstituted in benzyl benzoate (100 pL) and added to the syringe. The solution / syringe was then stored for 1 week at room temperature (2 samples) or 2-8°C (3 samples) in an upright position. After storage, the syringe was inverted and the solution was resuspended by swirling, and shaking or flicking. The injection plunger was attached, the cap was removed, and a 25 gauge needle was attached. The plunger was pressed and the contents dispensed into a beaker. The results showed no clogging of the needle, with a minimal compression force. See. FIG. 16.Example 6: Particle Size Measurements

[0236] Dry powders of anti-TLla Ab (samples A and B) were analyzed using high-resolution scanning electron microscopy (SEM). Sample A was dehydrated using 100% 1-pentanol as described herein and sieved through 75 micron mesh. Sample B was dehydrated using 30% ethyl lactate / 1 -pentanol as described herein and sieved through 75 micron mesh. Table 24 shows the results obtained from the particle detection and measurement.

[0237] FIGs. 17 and 18 show typical field of view of spherical particles detected from sample A and B, respectively. FIG. 19 shows the edge and shape detection results from Sample A (left) and B (right). Size magnification scale bars are at the bottom left margin of each SEM image.

[0238] The results show that the use of 30% ethyl lactate / 1 -pentanol as the dehydration solvent result in particles having a more uniform surface.Example 7: Density Analysis

[0239] The densities of four anti-TLIA Ab samples dehydrated as described herein using 100% 1-pentanol (2 samples) and 30% ethyl lactate / 1 -pentanol (2 samples) were analyzed using the parameters summarized in Table 25.

[0240] Table 25 provides the results of the density analyses.

[0241] These data show that average densities were obtained.Example 8: Break Loose and Glide / Extrusion Force Measurements

[0242] Three samples of anti -TL la Ab were analyzed for break loose and extrusion using a naked 1 mL long syringe (Ompi Nexa) with a stopper (West Novapure) at rate of 5 rnm / s. Samples were prepared by dehydrated anti-TLIA Ab using 30 % ethyl lactate / 1 -pentanol as described herein and passed through a 75 pM sieve. The samples were high concentration suspensions that were added to the syringes immediately after preparation. Syringes were then stopped and dispensed at 22°C using a 75 micron filter sieve (Post) at rate of 5 mm / s. Results are shown in Table 27.

[0243] These data show that the highly concentrated Ab solutions completely expelled from the syringe without clogging. See, FIG. 20 which shows clean injection plots for both concentrations.

[0244] Those skilled in the art will appreciate that numerous changes and modifications can be made to the preferred embodiments disclosed herein and that such changes and modifications can be made without departing from the spirit of the invention. It is, therefore, intended that the appended claims cover all such equivalent variations as fall within the true spirit and scope of the invention.

[0245] The disclosures of each patent, patent application, and publication cited or described in this document are hereby incorporated herein by reference, in its entirety.

Claims

What is claimed is:1 . A process for dehydrating an antibody-containing formulation, the process comprising: introducing the antibody-containing formulation into a dehydrating solvent mixture comprising 1 -pentanol and a cosolvent that is an ester, wherein the v / v ratio of the 1 -pentanol to the cosolvent is about 70 to about 90: about 30 to about 10, to thereby dehydrate the antibody-containing formulation.

2. The process of claim 1, wherein the ester is an acetate or lactate.

3. The process of claim 1 or 2, wherein the ester is an acetate.

4. The process of claim 3, wherein the acetate is ethyl acetate, butyl acetate, heptyl acetate, octyl acetate, or benzyl acetate, such as benzyl acetate.

5. The process of claim 1 or 2, wherein the ester is a lactate.

6. The process of claim 5, wherein the lactate is ethyl lactate, such as ethyl D-lactate, ethyl L-lactate, or ethyl D,L-lactate.

7. The process of any one of the preceding claims, wherein the ratio of the 1 -pentanol to the ester is about 90: about 10, about 80: about 20, or about 70: about 30.

8. The process of any one of the preceding claims, wherein the dehydrating solvent comprises 1 -pentanol and ethyl lactate.

9. The process of any one of the preceding claims, wherein the dehydrated antibodycontaining formulation comprises antibody-containing microbeads.

10. The process of claim 9, wherein the antibody-containing microbeads have an average diameter of less than about 30 pm, such as about 1 pm to about 20 pm.

11. The process of any one of the preceding claims, wherein the introducing is performed using homogenization, a Shirasu porous glass membrane, or microfluidics.

12. The process of any one of the preceding claims, wherein the introducing is performed at a rate of less than 50 pL / min.

13. The process of any one of the preceding claims, wherein the antibody-containing formulation further comprises sucrose.

14. The process of any one of the preceding claims, further comprising separating a liquid phase comprising the dehydrating solvent from a solid phase comprising the dehydrated antibody-containing formulation.

15. The process of claim 14, further comprising removing the liquid phase.

16. The process of claim 14, further comprising washing the solid phase.

17. The process of claim 16, wherein the washing is performed using ethanol, such as 200 proof ethanol.

18. The process of any one of the preceding claims, further comprising resuspending the dehydrated antibody-containing formulation in a vehicle to generate a suspension comprising antibody-containing microbeads.

19. The process of claim 18, wherein the vehicle is benzyl benzoate, benzyl alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of caprylic and capric acid, or combinations thereof.

20. The process of claim 18, wherein the vehicle is benzyl benzoate.

21. The process of any one of claims 18-20, wherein the suspension comprises less than about 0.5 area%, based on the area% of the antibody in the antibody-containing formulation prior to the dehydration, of dimerized antibody.

22. A dehydrated antibody-containing formulation prepared according to the process of any one of claims 1-21.

23. The dehydrated antibody-containing formulation of claim 22, wherein the antibody comprises an anti-IL-5 antibody. anti-IL-23 antibody. anti-PD-1 antibody, an anti- TLla antibody, an anti-CGRP antibody, or an anti-PAR2 antibody.

24. A suspension prepared according to the process of any one of claims 18-21.

25. A dosage form comprising the suspension of claim 24.

26. The dosage form of claim 25, which is a subcutaneous dosage form.

27. The dosage form of claim 25 or 26, wherein the dosage form comprises antibodycontaining microbeads.

28. The dosage form of claim 27, wherein the concentration of the antibody in the antibody-containing microbead suspension is at least about 150 mg / mL, such as up to about 400 mg / mL.

29. A suspension comprising: a dehydrating solvent comprising 1 -pentanol and a cosolvent that is an ester, wherein the v / v ratio of the 1-pentanol to the cosolvent is about 70 to about 90: about 30 to about 10, and an antibody-containing formulation.

30. The suspension of claim 29, wherein the v / v ratio of the 1-pentanol to the cosolvent is about 70: about 30. or about 80: about 20. or about 90: about 10.

31. The suspension of claim 29 or 30, wherein the ester is an acetate or lactate.

32. The suspension of any one of claims 29-31, wherein the dehydrating solvent comprises 1-pentanol and ethyl lactate.

33. The suspension of any one of claims 29-32, wherein at least a portion of the antibodycontaining formulation is in a form of antibody-containing microbeads.

34. A composition comprising: a vehicle and a plurality of antibody -containing microbeads, wherein the composition has one or more of the following characteristics as compared to a control composition comprising antibody-containing microbeads formed using 100% 1- pentanol: an injection force of about 2 to about 20 N when sieved with a 75 pm sieve, suspended in benzyl benzoate, and injected through a 27 G manual prefilled syringe (PFS);about 60% total weight of the material being < 75 microns (usable material) as compared to about 85% total weight of the material being > 75 microns (unusable material) when formed using 100% 1 -pentanol; a reduced settling of particles (about 80 % weight of particles formed using 100% 1 -pentanol are >75 micron and settle in about 2 minutes or less as compared to about 40 % weight of particles formed using ethyl lactate and 1 -pentanol which are >75 micron; a reduced settling (higher sedimentation rate) compared to a control composition as exemplified in FIG. 9; a greater particle uniformity as compared to particles formed using 100% 1- pentanol (75-micron sieved powder showed an increase of greater than 50% in particles with fractures and flocculations when dehydrated with 100% 1- pentanol alone; a greater powder uniformity7as compared to particles formed using 100% 1- pentanol (94% of particles in a 75-micron sieved powder were within a 30 pm range as compared to 87% of particles in a 75-micron sieved powder formed using 100% 1 -pentanol alone); smaller 75-micron sieved particle sizes as compared to particles formed using 100% 1 -pentanol (particle sizes of 15 um as compared to particle sizes of 25 um when formed using 100% 1 -pentanol); or a reduced percentage of flocculation as compared to the control composition as exemplified in FIGs. 3 and 4.

35. The composition of claim 34, wherein the antibody -containing microbeads have an average diameter of less than about 30 pm, such as about 1 to about 20 pm.

36. The composition of claim 34 or 35, wherein the vehicle is benzy l benzoate, benzy l alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of capry lic and capric acid, or combinations thereof.

37. A process for forming a high concentration antibody suspension, the process comprising:dehydrating an antibody-containing formulation according to the process of any one of claims 1-17 to form a dehydrated antibody -containing formulation; and resuspending the dehydrated antibody-containing formulation in a vehicle to form the high concentration antibody suspension.

38. The process of claim 37, wherein the dehydrated antibody-containing formulation comprises antibody-containing microbeads.

39. The process of claim 38, wherein the antibody-containing microbeads have an average diameter of less than about 30 pm, such as about 1 to about 20 pm.

40. The process of any one of claims 37-39, wherein the vehicle is benzyl benzoate, benzyl alcohol, ethyl alcohol, medium chain triglyceride, ethyl lactate, ethyl oleate, polyethylene glycol 200, propylene glycol, perfluoroalkane such as perfluorohexyloctane, perfluorodecane, perfluorooctane, or perfluorobutylpentane, isopropyl myristate, triglyceride of caprylic and capric acid, or combinations thereof.

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