High-concentration anti-C5 antibody preparation

Abstract

This disclosure provides a stable aqueous solution containing a high concentration of anti-C5 antibody (e.g., ravulizumab), and a method for preparing said aqueous solution. [Solution] The disclosure further provides a method for treating or preventing complement-related disorders, such as paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic uremic syndrome (aHUS), using the solution. The disclosure also features a therapeutic kit comprising one or more of the solution and means for administering the solution to a patient in need of such treatment.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the benefits of U.S. Provisional Patent Application No. 62 / 537,741, filed on 27 July 2017. The entire contents of the aforementioned Provisional Patent Application are incorporated herein by reference.

[0002] Sequence List This application includes a sequence listing, which has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. This ASCII copy was created on 26 July 2018, is named AXJ-226PC_SL.txt, and has a size of 32,987 bytes. [Background technology]

[0003] The complement system, working in conjunction with the body's other immune systems, defends against the invasion of cellular and viral pathogens. At least 25 complement proteins exist, presenting as a collection of complexes of plasma proteins and membrane cofactors. Plasma proteins account for approximately 10% of globulins in vertebrate serum. Complement components achieve their immunodefense functions by interacting in a complex but precise series of enzymatic cleavage and membrane binding events. The resulting complement cascade leads to the production of products with opsonizing, immunomodulatory, and lytic effects. A concise summary of the biological activities associated with complement activation can be found, for example, in The Merck Manual, 16. th It is offered in Edition.

[0004] While a properly functioning complement system provides stable defense against infectious microorganisms, when the complement system is inappropriately controlled or activated, it is involved in the pathogenesis of various disorders, including paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic uremic syndrome (aHUS) (for example, Socie G, et al., French Society of Haematology. Lancet. 1996; 348(9027):573 - 577; Brodsky, R., Blood. 2014; 124(18):2804 - 2811); Hillmen, P., et al, Am. J. Hematol. 2010; 85(8):553 - 559; Caprioli et al. (2006) Blood 108 : 1267 - 1279; and see Kavanagh et al. (2006) British Medical Bulletin 77 and 78:5 - 22).

[0005] Patients with complement - related disorders such as PNH or aHUS were at risk of significant morbidity and mortality. Accordingly, an object of the present invention is to provide improved compositions and methods for treating patients having complement - related disorders.

Prior Art Documents

Non - Patent Documents

[0006]

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[0007] This specification provides stable, high-concentration aqueous solutions of anti-C5 antibodies, as well as methods for preparing and using such formulations. In particular, this disclosure provides formulation conditions suitable for maintaining the long-term physical and functional stability of anti-C5 antibodies (e.g., ravulizumab, also known as "antibody BNJ441" and "ALXN1210") in high-concentration solutions. For example, this disclosure provides formulation conditions that allow anti-C5 antibodies to be maintained, primarily in monomeric form, for up to two years at 2°C to 8°C, even when the antibody is maintained in a solution at a concentration of approximately 100 mg / mL or higher. Furthermore, as described herein and illustrated in the examples, such formulations also minimize aggregation, fragmentation, or degradation of anti-C5 antibodies (e.g., ravulizumab) in high-concentration solutions. For example, this disclosure provides formulation conditions that allow anti-C5 antibodies to be maintained at high concentrations for two years without detection of antibody fragmentation or degradation products (when measured using size exclusion chromatography-high-performance liquid chromatography (SEC-HPLC), e.g., HPLC-gel permeation), and with aggregation of 2% or less. Furthermore, this specification provides conditions suitable for the formulation of anti-C5 antibody solutions, such as ravulizumab, at concentrations exceeding 200 mg / mL.

[0008] Stable, high-concentration anti-C5 antibody solutions offer numerous advantages. Firstly, in therapeutic applications requiring the administration of antibodies in small volumes, therapeutic efficacy often depends on the amount of antibody that can be delivered in that small volume. If high-concentration anti-C5 antibodies cannot be formulated, routes such as subcutaneous, intravitreous, and / or intra-articular delivery are often excluded. In this regard, high-concentration antibody formulations offer patients more options regarding the route of administration. For therapeutic applications requiring frequent and chronic administration, and / or self-delivery, administration with high-concentration formulations makes this possible and would be more preferable to intravenous infusion for patients. For example, high-concentration anti-C5 antibody formulations would allow patients to self-administer antibodies, such as by subcutaneous or intravenous injection. Therefore, formulating antibodies at high concentrations provides a convenient alternative for home administration, increasing administration compliance in patients with complement-related disorders.

[0009] Furthermore, the method for preparing the aqueous solution described herein does not require a lyophilization step, nor does it require reconstitution of the high-concentration aqueous solution from the lyophilized material. The high-concentration antibody solution offers several advantages over reconstituted lyophilized antibody preparations. Firstly, physicians must reconstitute the lyophilized antibody solution aseptically in the field, but this process increases the chance of bacterial contamination of the solution before administration. Furthermore, reconstitution requires careful dissolution of all solids contained in the reconstitution tube into the solution. Thus, the high-concentration aqueous solution provided herein provides physicians, caregivers, and / or patients with a means of delivering therapeutic antibodies rapidly, simply, safely, and efficiently to patients in need.

[0010] Other advantages of high-concentration formulations include, for example, reduced bulk storage space and / or lower manufacturing costs due to the reduced number of products required. Furthermore, producing products with longer shelf lives ultimately reduces the total production volume needed. This ultimately reduces costs for both manufacturers and consumers of high-concentration therapeutic antibodies.

[0011] Exemplary anti-C5 antibodies are ravulizumab (also known as the antibodies BNJ441 and ALXN1210), or its antigen-binding fragments and variants, comprising heavy and light chains having sequences shown in SEQ ID NOs. 14 and 11, respectively. In other embodiments, the antibody comprises the complementarity-determining regions (CDRs) or variable regions (VRs) of the heavy and light chains of ravulizumab. Thus, in one embodiment, the antibody comprises the CDR1, CDR2, and CDR3 domains of the heavy chain variable (VH) region of ravulizumab having the sequence described in SEQ ID NOs. 12, and the CDR1, CDR2, and CDR3 domains of the light chain variable (VL) region of ravulizumab having the sequence described in SEQ ID NOs. 8. In another embodiment, the antibody comprises the heavy chain sequences of CDR1, CDR2, and CDR3 described in SEQ ID NOs. 19, 18, and 3, respectively, and the light chain sequences of CDR1, CDR2, and CDR3 described in SEQ ID NOs. 4, 5, and 6, respectively.

[0012] In another embodiment, the antibody includes a VH region and a VL region having the amino acid sequences described in SEQ ID NO: 12 and SEQ ID NO: 8, respectively.

[0013] In another embodiment, the antibody includes a heavy chain constant region as described in SEQ ID NO: 13.

[0014] In another embodiment, the antibody comprises a variant human Fc constant region that binds to the human neonatal Fc receptor (FcRn), wherein the variant human Fc CH3 constant region comprises substitutions of Met-429-Leu and Asn-435-Ser, respectively, in EU numbering, at the residues corresponding to methionine 428 and asparagine 434 in the native human IgG Fc constant region.

[0015] In another embodiment, the antibody comprises the heavy chain sequences of CDR1, CDR2, and CDR3 described in SEQ ID NOs. 19, 18, and 3, respectively, and the light chain sequences of CDR1, CDR2, and CDR3 described in SEQ ID NOs. 4, 5, and 6, respectively, as well as a variant human Fc constant region that binds to the human neonatal Fc receptor (FcRn), wherein the variant human Fc CH3 constant region, in EU numbering, includes substitutions of Met-429-Leu and Asn-435-Ser at the residues corresponding to methionine 428 and asparagine 434 in the native human IgG Fc constant region.

[0016] In another embodiment, the antibody competes with the antibody described above for binding and / or binds to the same C5 epitope as the antibody described above. In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity with the antibody described above (e.g., at least about 90%, 95%, or 99% variable region identity with SEQ ID NO: 12 and SEQ ID NO: 8).

[0017] In another embodiment, the antibody is 0.1 nM ≤ K at pH 7.4 and 25°C. D Affinity dissociation constant (K) in the range ≤ 1 nM D ) binds to human C5. In another embodiment, the antibody is at pH 6.0 and 25°C, K D The antibody binds to human C5 at a concentration of ≥10 nM. In yet another embodiment, the antibody or its antigen-binding fragment against human C5 at pH 6.0 and 25°C is K D ) / (K of antibody or antigen-binding fragment against human C5 at pH 7.4 and 25°C) D )] is greater than 25.

[0018] In one embodiment, a stable aqueous solution (e.g., a sterile solution) is provided, the solution containing an anti-C5 antibody at a concentration of about 100 mg / mL, the anti-C5 antibody comprising a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6. In another embodiment, the solution contains an anti-C5 antibody (e.g., ravulizumab) at a concentration of approximately 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, or 300 mg / mL.

[0019] In another embodiment, the stable aqueous solution comprises one or more additional agents (e.g., stabilizers, buffers, surfactants, and / or preservatives). For example, in one embodiment, the stable aqueous solution comprises a stabilizer. Exemplary stabilizers include, but are not limited to, polyols, sugars (e.g., sucrose or trehalose), amino acids (e.g., arginine), amines, and salting-out salts. In one embodiment, the solution comprises both ends and contains at least one stabilizer at a concentration of 2–10%. In one embodiment, the solution comprises both ends and contains at least one stabilizer at a concentration of 10 mM–50 mM. In another embodiment, the stabilizer is present in the solution at a concentration of 20 mM or higher. In another embodiment, the stabilizer is present in the solution at a concentration of at least 25 mM or 25 mM. In another embodiment, the stabilizer is present in the solution at a concentration of 50 mM or higher. In another embodiment, the solution contains 25 mM arginine.

[0020] In another embodiment, the solution comprises at least one buffer. Typical buffers that may be contained in the washing solution include: Tris(tris(hydroxymethyl)methylamine), bis-tris, bis-trispropane, histidine, triethanolamine, diethanolamine, formate, acetate, MES(2-(N-morpholino)ethanesulfonic acid), phosphate, HEPES(4-2-hydroxyethyl-1-piperazineethanesulfonic acid), citrate, MOPS(3-(N-morpholino)propanesulfonic acid), TAPS(3{[tris(hydroxymethyl)methyl]amino}propanesulfonic acid), Bicine(N,N-bis(2-hydroxyethyl)glycine), Tricine(N-tris( Examples include hydroxymethyl(methylglycine), TES (2-{[tris(hydroxymethyl)methyl]amino}ethanesulfonic acid), PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid), cacodylate (dimethylarsinic acid), SSC (sodium citrate salt), and sodium phosphate. In another embodiment, the buffer is an amino acid. The amino acid may be an amino acid selected from the group consisting of, for example, histidine (e.g., L-histidine), serine (e.g., L-serine), and glycine (e.g., L-glycine). In another embodiment, the solution contains two or more buffers. In a particular embodiment, the buffer is sodium phosphate.

[0021] In another embodiment, the solution includes both ends and contains at least one buffer at a concentration of 10 mM to 300 mM. In another embodiment, the solution includes both ends and contains at least one buffer at a concentration of 10 mM to 200 mM. In another embodiment, the solution includes both ends and contains at least one buffer at a concentration of 10 mM to 100 mM. In another embodiment, the solution includes both ends and contains at least one buffer at a concentration of 10 mM to 50 mM. In another embodiment, the solution includes both ends and contains at least one buffer at a concentration of 20 mM to 50 mM. In another embodiment, the buffer is present in the solution at a concentration of 20 mM or higher. In another embodiment, the buffer is present in the solution at a concentration of 25 mM or higher. In another embodiment, the buffer is present in the solution at a concentration of 50 mM or higher.

[0022] In another embodiment, the solution contains a sugar excipient at a concentration of 0.1 to 5%. In one embodiment, the sugar excipient is present in the solution at a concentration of 1.5% or more. In another embodiment, the sugar excipient is present in the solution at a concentration of 3% or more. The sugar excipient may be a sugar selected from the group consisting of, for example, sorbitol and mannitol. In another embodiment, the solution contains two or more sugar excipients.

[0023] In another embodiment, the solution contains a surfactant. Suitable surfactants for use in the formulations of the present invention include, but are not limited to, fatty acid esters (e.g., sorbitan monocaprate, sorbitan monolaurate, sorbitan monopalmitate), sorbitan trioleates, glycerin fatty acid esters (e.g., glycerin monocaprate, glycerin monomyristate, glycerin monostearate), polyglycerin fatty acid esters (e.g., decaglyceryl monostearate, decaglyceryl distearate, decaglyceryl monolinoleate), polyoxyethylene sorbitan fatty acid esters (e.g., sorbitan polyoxyethylene monolaurate, sorbitan polyoxyethylene monooleate, sorbitan polyoxyethylene monostearate, sorbitan polyoxyethylene monopalmitate, sorbitan polyoxyethylene trioleate, sorbitan polyoxyethylene tristearate), and polyoxyethylene sorbitol fatty acid esters (e.g., sorbitol polyoxyethylene tetrastearate, sorbitol polyoxyethylene tetraoleate). , polyoxyethylene glycerin fatty acid esters (e.g., glyceryl polyethylene monostearate), polyethylene glycol fatty acid esters (e.g., glycol polyethylene distearate), polyoxyethylene alkyl esters (e.g., polyoxyethylene lauryl ether), polyoxyethylene polyoxypropylene alkyl ethers (e.g., polyoxyethylene polyoxypropylene glycol, polyoxyethylene polyoxypropylene propyl ether, polyoxyethylene polyoxypropylene cetyl ether), polyoxyethylene alkylphenyl ethers (e.g., polyoxyethylene nonylphenyl ether), polyoxyethylene hydrogenated castor oil (e.g., polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil), polyoxyethylene beeswax derivatives (e.g., polyoxyethylene sorbitol beeswax), polyoxyethylene lanolin derivatives (e.g., polyoxyethylene lanolin), and polyoxyethylene fatty acid amides (e.g., polyoxyethylene stearic acid amide);Examples include C12-C18 alkyl sulfates (e.g., sodium cetyl sulfate, sodium lauryl sulfate, sodium oleyl sulfate), polyoxyethylene C10-C18 alkyl ether sulfates with an average of 2-4 moles of ethylene oxide units added (e.g., sodium polyoxyethylene lauryl sulfate), and C10-C18 alkyl sulfosuccinate salts (e.g., sodium lauryl sulfosuccinate); as well as natural surfactants such as lecithin, glycerophospholipids, and sphingophospholipids (e.g., sphingomyelin), and sucrose esters of C12-C18 fatty acids.

[0024] In one embodiment, the surfactant in the formulation is a nonionic surfactant. In a particular embodiment, the surfactant in the formulation is a polyoxyethylene sorbitan fatty acid ester, such as polysorbate 20, 40, 60, 80, or one or more combinations thereof. In one embodiment, the surfactant in the formulation is polysorbate 80 (Tween 80). In another embodiment, the surfactant in the formulation is polysorbate 60. In another embodiment, the surfactant in the formulation is polysorbate 40. In another embodiment, the surfactant in the formulation is polysorbate 20 (Tween 20). The concentration of the surfactant in the solution may be, for example, 0.001% to 0.02%, including both ends. For example, the surfactant may be present in the formulation in an amount of about 0.001% to about 1%, or about 0.001% to about 0.5%, or about 0.01% to about 0.2%. In one embodiment, the aqueous solution contains at least, or approximately, 0.001 (for example, at least, or approximately 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, Contains surfactants at concentrations of 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.3, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, or 0.5% or higher. In another embodiment, the aqueous solution contains 0.2% (e.g., 0.19, 0.18, 0.17, 0.16, 0.15, 0.14, 0.13, 0.12, 0.11, 0.10, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, or 0.001)% or less of a pharmaceutically acceptable surfactant. In a particular embodiment, the surfactant is 0.05% polysorbate 80.

[0025] In another embodiment, the solution contains a preservative. Examples of exemplary preservatives include, but are not limited to, benzyl alcohol, m-cresol, and phenol.

[0026] In one embodiment, the stable aqueous solution contains five or fewer agents in addition to the anti-C5 antibody. In another embodiment, the stable aqueous solution contains four or fewer agents in addition to the anti-C5 antibody. In yet another embodiment, the stable aqueous solution contains three or fewer agents in addition to the anti-C5 antibody. In yet another embodiment, the stable aqueous solution contains two or fewer agents in addition to the anti-C5 antibody. In yet another embodiment, the stable aqueous solution contains one or fewer agents in addition to the anti-C5 antibody.

[0027] In another embodiment, the stable aqueous solution contains a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and an amino acid sequence described in SEQ ID NO: 6. Anti-C5 antibody containing a light chain CDR3 with a minoic acid sequence, 50±15 mM (e.g., 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or 65) mM phosphate buffer, 5±3 (e.g., 2, 3, 4, 5, 6, 7 or 8) % sucrose , and also comprising 25±10 (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) mM of arginine, in which case the solution has a pH of 7.4±0.5 (e.g., 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8 or 7.9).

[0028] In another embodiment, the stable aqueous solution contains a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and an amino acid sequence described in SEQ ID NO: 6, all at a concentration of 100 ± 20 mg / mL (e.g., 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120) mg / mL, a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and an amino acid sequence described in SEQ ID NO: 6 Anti-C5 antibody containing a light chain CDR3 containing an acid sequence, 50±15 mM (e.g., 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or 65) mM phosphate buffer, 5±3 (e.g., 2, 3, 4, 5, 6, 7 or 8) % sucrose, The solution consists of 25 ± 10 mM (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) mM arginine, in which case the solution has a pH of 7.4 ± 0.5 (e.g., 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, or 7.9).

[0029] In another embodiment, the stable aqueous solution has a concentration of 100 ± 20 mg / mL (e.g., 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120) Anti-C5 antibody containing heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6, 50±15 (for example) 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or 65) mM phosphate buffer, 5 ± 3 (e.g., 2, 3, 4, 5, 6, 7 or 8) % sucrose, 25 ± 10 (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 2 The solution comprises 7, 28, 29, 30, 31, 32, 33, 34, or 35 mM arginine, and 0.05 ± 0.03 (e.g., 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, and 0.08)% polysorbate 80, in which case the solution has a pH of 7.4 ± 0.5 (e.g., 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, or 7.9).

[0030] In another embodiment, the stable aqueous solution is a mixture of 100 ± 20 mg / mL (e.g., 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120) mg / mL. Anti-C5 antibody containing heavy chain CDR1 containing the amino acid sequence described in sequence number 19, heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6, 50±15 (for example) 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or 65) mM phosphate buffer, 5 ± 3 (e.g., 2, 3, 4, 5, 6, 7 or 8) % sucrose, 25 ± 10 (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 The solution comprises 28, 29, 30, 31, 32, 33, 34 or 35 mM arginine and 0.05 ± 0.03 (e.g., 0.02, 0.03, 0.04, 0.05, 0.06, 0.07 and 0.08)% polysorbate 80, in which case the solution has a pH of 7.4 ± 0.5 (e.g., 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8 or 7.9).

[0031] In another embodiment, a stable aqueous solution is provided (e.g., a sterile solution) in which the solution comprises (a) an anti-C5 antibody (e.g., ravulizumab), (b) about 50 mM phosphate buffer, (c) about 5% sucrose, and (d) about 25 mM arginine. In another embodiment, a stable aqueous solution is provided (e.g., a sterile solution) in which the solution comprises (a) an anti-C5 antibody (e.g., ravulizumab) at a concentration of about 100 mg / mL, (b) about 50 mM phosphate buffer, (c) about 5% sucrose, and (d) about 25 mM arginine. In another embodiment, the stable aqueous solution comprises a) an anti-C5 antibody (e.g., ravulizumab), (b) 50 mM phosphate buffer, (c) 5% sucrose, and (d) 25 mM arginine. In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody (e.g., ravulizumab) at a concentration of 100 mg / mL, (b) 50 mM phosphate buffer, (c) 5% sucrose, and (d) 25 mM arginine.

[0032] In another embodiment, the stable aqueous solution comprises (a) anti-C5 antibody, (b) about 50 mM phosphate buffer, (c) about 5% sucrose, (d) about 0.05% polysorbate 80, and (e) 25 mM arginine. In another embodiment, the stable aqueous solution comprises (a) anti-C5 antibody at a concentration of about 100 mg / mL, (b) about 50 mM phosphate buffer, (c) about 5% sucrose, (d) about 0.05% polysorbate 80, and (e) about 25 mM arginine. In another embodiment, the stable aqueous solution comprises a) anti-C5 antibody, (b) 50 mM phosphate buffer, (c) 5% sucrose, (d) 0.05% polysorbate 80, and (e) 25 mM arginine.

[0033] In another embodiment, the stable aqueous solution comprises (a) anti-C5 antibody, (b) 50 mM phosphate buffer, (c) 5% sucrose, (d) 0.05% polysorbate 80, and (e) 25 mM arginine. In another embodiment, the stable aqueous solution comprises (a) anti-C5 antibody at a concentration of 100 mg / mL, (b) 50 mM phosphate buffer, (c) 5% sucrose, (d) 0.05% polysorbate 80, and (e) 25 mM arginine. In another embodiment, the stable aqueous solution comprises a) anti-C5 antibody at a concentration of 100 mg / mL, (b) 50 mM phosphate buffer, (c) 5% sucrose, (d) 0.05% polysorbate 80, and (e) 25 mM arginine.

[0034] In another embodiment, the stable aqueous solution contains four or fewer agents in addition to the anti-C5 antibody. In another embodiment, the stable aqueous solution contains three or fewer agents in addition to the anti-C5 antibody. In another embodiment, the stable aqueous solution contains two or fewer agents in addition to the anti-C5 antibody. In another embodiment, the stable aqueous solution contains one or fewer agents in addition to the anti-C5 antibody.

[0035] In another embodiment, the stable aqueous solution consists of (a) an anti-C5 antibody at a concentration of about 100 mg / mL, (b) a phosphate buffer at about 50 mM, (c) sucrose at about 5%, and (d) arginine at about 25 mM.

[0036] In another embodiment, the stable aqueous solution consists of (a) an anti-C5 antibody at a concentration of about 100 mg / mL, (b) about 50 mM phosphate buffer, (c) about 5% sucrose, (d) about 0.05% polysorbate 80, and (e) about 25 mM arginine.

[0037] In one embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of about 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) about 50 mM phosphate buffer; (c) about 5% sucrose; and (d) about 25 mM arginine.

[0038] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) 50 mM phosphate buffer; (c) 5% sucrose; and (d) 25 mM arginine.

[0039] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of about 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) about 50 mM phosphate buffer; (c) about 5% sucrose; (d) about 0.05% polysorbate 80; and (e) about 25 mM arginine.

[0040] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) 50 mM phosphate buffer; (c) 5% sucrose; (d) 0.05% polysorbate 80; and (e) about 25 mM arginine.

[0041] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of about 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) about 50 mM phosphate buffer; (c) about 5% sucrose; (d) about 0.05% polysorbate 80; and (e) about 25 mM arginine.

[0042] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) 50 mM phosphate buffer; (c) 5% sucrose; (d) 0.05% polysorbate 80; and (e) 25 mM arginine.

[0043] In one embodiment, the pH is 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, or 7.9. In another embodiment, the pH of the solution is 7.0 to 7.4. In yet another embodiment, the pH of the solution is 7.2 to 7.8. In yet another embodiment, the pH of the solution is 7.2 to 7.6. In a particular embodiment, the pH of the solution is 7.4.

[0044] The solutions described herein may be formulated for any suitable mode of administration. In one embodiment, the solution is formulated for parenteral administration (e.g., intravenous, subcutaneous, intraperitoneal, or intramuscular injection). In a particular embodiment, the solution is formulated for subcutaneous administration. For example, in one embodiment, a stable aqueous solution containing an anti-C5 antibody at a concentration of 100 mg / mL is formulated for subcutaneous administration. In another particular embodiment, the solution is formulated for intravenous administration. For example, in one embodiment, a stable aqueous solution containing an anti-C5 antibody at a concentration of 100 mg / mL is formulated for intravenous administration.

[0045] In one embodiment of any of the solutions described herein, the anti-C5 antibody (e.g., ravulizumab) retains at least 95% (e.g., at least 96, 97, 98, or 99%) monomers during a storage period of at least 6 months at 2°C to 8°C, as determined by SEC-HPLC (e.g., gel permeation HPLC). In another embodiment, the anti-C5 antibody retains at least 95% (e.g., at least 96, 97, 98, or 99%) monomers during a storage period of at least 9 months at 2°C to 8°C, as determined by SEC-HPLC. In yet another embodiment, the anti-C5 antibody retains at least 95% (e.g., at least 96, 97, 98, or 99%) monomers during a storage period of at least 1 year at 2°C to 8°C, as determined by SEC-HPLC. In yet another embodiment, the anti-C5 antibody retains at least 95% (e.g., at least 96, 97, 98, or 99%) monomers during a storage period of at least 18 months at 2°C to 8°C, as determined by SEC-HPLC. In another embodiment, the anti-C5 antibody retains at least 95% (e.g., at least 96, 97, 98, or 99%) of monomers during a storage period of at least two years at 2°C to 8°C, as determined by SEC-HPLC.

[0046] In any other embodiment of the solutions described herein, less than 5% of the anti-C5 antibody (e.g., ravulizumab) in the solution is aggregated when determined by SEC-HPLC (e.g., gel permeation HPLC). In another embodiment, less than 4% of the anti-C5 antibody in the solution is aggregated when determined by SEC-HPLC. In another embodiment, less than 3% of the anti-C5 antibody in the solution is aggregated when determined by SEC-HPLC. In another embodiment, less than 2% of the anti-C5 antibody in the solution is aggregated when determined by SEC-HPLC. In another embodiment, less than 1% of the anti-C5 antibody in the solution is aggregated when determined by SEC-HPLC.

[0047] In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains its C5-binding activity by at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) for at least 6 months at 2°C to 8°C compared to a reference anti-C5 antibody corresponding to the anti-C5 antibody before storage. In another embodiment, an anti-C5 antibody (e.g., ravulizumab) maintains its C5-binding activity by at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) for at least 9 months at 2°C to 8°C compared to a reference anti-C5 antibody corresponding to the anti-C5 antibody before storage. In another embodiment, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99)% of its C5-binding activity for at least 18 months at 2°C to 8°C, compared to a reference anti-C5 antibody (e.g., ravulizumab) that corresponds to an anti-C5 antibody before storage. In another embodiment, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) of its C5-binding activity for at least 3 years at 2°C to 8°C, compared to a reference anti-C5 antibody (e.g., ravulizumab) that corresponds to an anti-C5 antibody before storage.

[0048] In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99)% of its hemolytic inhibitory ability for at least 6 months at 2°C to 8°C, compared to a reference anti-C5 antibody (e.g., ravulizumab) that corresponds to an anti-C5 antibody before storage. In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99)% of its hemolytic inhibitory capacity for at least 18 months at 2°C to 8°C, compared to a reference anti-C5 antibody (e.g., ravulizumab) that corresponds to an anti-C5 antibody before storage. In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99)% of its hemolytic inhibitory capacity for at least 18 months at 2°C to 8°C, compared to a reference anti-C5 antibody that corresponds to an anti-C5 antibody before storage. In any other embodiment of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) of its hemolytic inhibitory ability for at least two years at 2°C to 8°C, compared to a reference anti-C5 antibody equivalent to the anti-C5 antibody before storage.In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) of its hemolytic inhibitory ability for at least 3 years at 2°C to 8°C, compared to a reference anti-C5 antibody equivalent to the anti-C5 antibody before storage.

[0049] In another embodiment, a method is provided for preparing a stable concentrated antibody solution comprising 100 mg / mL of anti-C5 antibody, 50 mM phosphate buffer, 5% sucrose, and 25 mM arginine, the method being

[0050] i) To provide a first aqueous solution containing an anti-C5 antibody, wherein the first aqueous solution has a first formulation and contains an anti-C5 antibody at a concentration of 10 mg / mL or less.

[0051] ii) Dialysis filtration is performed on the first aqueous solution to obtain a formulation containing 50 mM phosphate buffer, 5% sucrose, and 25 mM arginine at pH 7.4, thereby producing a second aqueous solution, wherein the second aqueous solution has the second formulation as a result of the dialysis filtration, and

[0052] iii) The second aqueous solution is concentrated to prepare a stable concentrated antibody solution containing 100 mg / mL of anti-C5 antibody, 50 mM phosphate buffer, 5% sucrose, and 25 mM arginine.

[0053] In another embodiment, a method is provided for preparing a stable concentrated antibody solution comprising 100 mg / mL of anti-C5 antibody, 50 mM phosphate buffer, 5% sucrose, 25 mM arginine, and 0.05% polysorbate 80, the method being described as follows:

[0054] i) To provide a first aqueous solution containing an anti-C5 antibody, wherein the first aqueous solution has a first formulation and contains an anti-C5 antibody at a concentration of 10 mg / mL or less.

[0055] ii) Dialysis filtration is performed on the first aqueous solution to a formulation containing 50 mM phosphate buffer, 5% sucrose, 25 mM arginine, and 0.05% polysorbate 80 at pH 7.4, thereby producing a second aqueous solution, wherein the second aqueous solution has the second formulation as a result of the dialysis filtration, and

[0056] iii) The second aqueous solution is concentrated to prepare a stable concentrated antibody solution containing 100 mg / mL of anti-C5 antibody, 50 mM phosphate buffer, 5% sucrose, 25 mM arginine, and 0.05% polysorbate 80.

[0057] A method for treating human patients with complement-related conditions is also provided, comprising administering to the patient (e.g., subcutaneously or intravenously) a stable aqueous solution described herein in an amount effective to treat the complement-related condition. Examples of complement-related conditions, but not limited to, include rheumatoid arthritis, antiphospholipid syndrome, lupus nephritis, ischemia-reperfusion injury, atypical hemolytic uremic syndrome (aHUS), typical hemolytic uremic syndrome, paroxysmal nocturnal hemoglobinuria (PNH), dense deposit disease, neuromyelitis optica, multifocal motor neuropathy, multiple sclerosis, macular degeneration, HELLP syndrome, spontaneous fetal disappearance, and thrombotic thrombocytopenic purpura. Plaque disease, pauci-immune type vasculitis, epidermolysis bullosa, recurrent miscarriage, traumatic brain injury, myocarditis, cerebrovascular disease, peripheral vascular disease, renal vascular disease, mesenteric / intestinal vascular disease, vasculitis, Henoch-Schönlein purpura nephritis, lupus erythematosus-associated vasculitis, vasculitis associated with rheumatoid arthritis, immune complex vasculitis, Takayasu's arteritis, dilated cardiomyopathy, diabetic vasculopathy, Kawasaki disease, venous gas embolism (venous gas embolism) Examples of complement-related conditions include gas embolus, restenosis after stent placement, plaque resection, percutaneous transluminal coronary angioplasty, myasthenia gravis, cold agglutinin disease, dermatomyositis, paroxysmal cold hemoglobinuria, antiphospholipid syndrome, Graves' disease, atherosclerosis, Alzheimer's disease, sepsis with systemic inflammatory response, septic shock, spinal cord injury, glomerulonephritis, graft rejection, Hashimoto's thyroiditis, type 1 diabetes mellitus, psoriasis, pemphigus, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, Goodpasture syndrome, Degos disease, and fulminant antiphospholipid syndrome. In certain embodiments, the complement-related condition is atypical hemolytic uremic syndrome (aHUS). In other embodiments, the complement-related condition is paroxysmal nocturnal hemoglobinuria (PNH).

[0058] Furthermore, a kit is provided containing a therapeutically effective amount of the stable aqueous solution described herein, adapted for use in the methods described herein. In one embodiment, the kit comprises (i) any of the solutions described herein, and (ii) means (e.g., a syringe) for delivering the solution to a patient in need. In one embodiment, the means is suitable for subcutaneous delivery of the solution to the patient. In one embodiment, the means is suitable for intravenous delivery of the solution to the patient. In another embodiment, the kit further comprises at least one additional activator for use in the treatment of complement-related conditions in a subject. The present invention provides, for example, the following items: (Item 1) It is a stable aqueous solution, (a) An anti-C5 antibody with a concentration of approximately 100 mg / mL, comprising: heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19; heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18; heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3; light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4; light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5; and light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6. (b) Approximately 50 mM phosphate buffer, (c) Approximately 5% sucrose, and (d) A stable aqueous solution containing approximately 25 mM arginine. (Item 2) It is a stable aqueous solution, (a) An anti-C5 antibody comprising a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6. (b) Approximately 50 mM phosphate buffer, (c) Approximately 5% sucrose, (d) Approximately 0.05% polysorbate 80, and (e) A stable aqueous solution containing approximately 25 mM arginine. (Item 3) A stable aqueous solution as described in item 1, further containing a surfactant. (Item 4) The stable aqueous solution described in item 3, wherein the surfactant is approximately 0.05% polysorbate 80. (Item 5) The stable aqueous solution of the anti-C5 antibody described above, at a concentration of approximately 100 mg / mL, as described in item 2. (Item 6) The stable aqueous solution according to any one of items 1 to 5, further comprising a variant human Fc constant region that binds to the human neonatal Fc receptor (FcRn), wherein the variant human Fc CH3 constant region comprises substitutions of Met-429-Leu and Asn-435-Ser in EU numbering, respectively, at the residues corresponding to methionine 428 and asparagine 434 of the natural human IgG Fc constant region. (Item 7) A stable aqueous solution according to any one of items 1 to 6, wherein the anti-C5 antibody comprises a heavy chain variable region described in SEQ ID NO: 12 and a light chain variable region described in SEQ ID NO: 8. (Item 8) The anti-C5 antibody is a stable aqueous solution according to any one of items 1 to 7, wherein the anti-C5 antibody contains the heavy chain constant region described in SEQ ID NO: 13. (Item 9) The stable aqueous solution according to any one of items 1 to 8, wherein the anti-C5 antibody comprises a heavy chain polypeptide having the amino acid sequence described in SEQ ID NO: 14 and a light chain polypeptide having the amino acid sequence described in SEQ ID NO: 11. (Item 10) A stable aqueous solution according to any one of items 1 to 9, wherein the anti-C5 antibody is ALXN1210 (ravulizumab). (Item 11) A stable aqueous solution according to any one of items 1 to 10, wherein the pH of the solution is 7.2 to 7.6. (Item 12) The stable aqueous solution described in item 11, wherein the pH of the solution is 7.4. (Item 13) The aforementioned solution is a sterilized, stable aqueous solution as described in any one of items 1 to 12. (Item 14) A stable aqueous solution according to any one of items 1 to 13, wherein the anti-C5 antibody maintains at least 97% monomer content when determined by SEC-HPLC during storage at 2°C to 8°C for at least 6 months. (Item 15) A stable aqueous solution according to any one of items 1 to 14, wherein the anti-C5 antibody maintains at least 97% monomer content when determined by SEC-HPLC during storage at 2°C to 8°C for at least one year. (Item 16) A stable aqueous solution according to any one of items 1 to 15, wherein less than 3% of the anti-C5 antibody in the solution is aggregated as determined by SEC-HPLC. (Item 17) A stable aqueous solution according to any one of items 1 to 16, wherein less than 2% of the anti-C5 antibody in the solution is aggregated as determined by SEC-HPLC. (Item 18) A stable aqueous solution according to any one of items 1 to 17, wherein less than 1% of the anti-C5 antibody in the solution is aggregated as determined by SEC-HPLC. (Item 19) A stable aqueous solution according to any one of items 1 to 18, wherein, during storage at 2°C to 8°C for at least 6 months, the anti-C5 antibody maintains at least 90% of its C5 binding activity compared to the anti-C5 antibody before storage. (Item 20) A stable aqueous solution according to any one of items 1 to 19, wherein, during storage at 2°C to 8°C for at least 6 months, the anti-C5 antibody maintains at least 95% of its hemolytic inhibitory ability compared to the anti-C5 antibody before storage. (Item 21) The aforementioned solution is a stable aqueous solution according to any one of items 1 to 20, which is suitable for subcutaneous administration. (Item 22) The aforementioned solution is a stable aqueous solution as described in any one of items 1 to 21, suitable for intravenous administration. (Item 23) A method for preparing a stable concentrated antibody solution comprising an anti-C5 antibody at a concentration of 100 mg / mL, containing a heavy chain CDR1 having the amino acid sequence described in SEQ ID NO: 23, a heavy chain CDR2 having the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR3 having the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 having the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 having the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 having the amino acid sequence described in SEQ ID NO: 6, 50 mM phosphate buffer, 5% sucrose, and 25 mM arginine, i) To provide a first aqueous solution containing the anti-C5 antibody, wherein the first aqueous solution has a first formulation and contains the anti-C5 antibody at a concentration of 10 mg / mL or less. ii) Dialysis filtration is performed on the first aqueous solution to obtain a formulation containing 50 mM phosphate buffer, 5% sucrose, and 25 mM arginine at pH 7.4, thereby producing a second aqueous solution, wherein the second aqueous solution has the second formulation as a result of the dialysis filtration. iii) A method comprising concentrating the second aqueous solution to prepare a stable concentrated antibody solution containing 100 mg / mL of anti-C5 antibody, 50 mM phosphate buffer, 5% sucrose, and 25 mM arginine. (Item 24) A method for treating a human patient having a complement-related condition, comprising administering to the patient a stable aqueous solution described in any one of items 1 to 21 in an amount effective for treating the complement-related condition. (Item 25) The complement-related conditions include rheumatoid arthritis, antiphospholipid syndrome, lupus nephritis, ischemia-reperfusion injury, atypical hemolytic uremic syndrome (aHUS), typical hemolytic uremic syndrome, paroxysmal nocturnal hemoglobinuria (PNH), dense deposit disease, neuromyelitis optica, multifocal motor neuropathy, multiple sclerosis, macular degeneration, HELLP syndrome, spontaneous fetal disappearance, thrombotic thrombocytopenic purpura, and vascular Pauci-immune type inflammation, epidermolysis bullosa, recurrent miscarriage, traumatic brain injury, myocarditis, cerebrovascular disease, peripheral vascular disease, renal vascular disease, mesenteric / intestinal vascular disease, vasculitis, Henoch-Schönlein purpura nephritis, lupus erythematosus-associated vasculitis, vasculitis associated with rheumatoid arthritis, immune complex vasculitis, Takayasu's arteritis, dilated cardiomyopathy, diabetic vasculopathy, Kawasaki disease, venous gas embolism. The method described in item 23, selected from the group consisting of gas embolus, restenosis after stent placement, atherosclerosis, percutaneous transluminal coronary angioplasty, myasthenia gravis, cold agglutinin disease, dermatomyositis, paroxysmal cold hemoglobinuria, antiphospholipid syndrome, Graves' disease, atherosclerosis, Alzheimer's disease, systemic inflammatory septicemia, septic shock, spinal cord injury, glomerulonephritis, graft rejection, Hashimoto's thyroiditis, type 1 diabetes mellitus, psoriasis, pemphigus, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, Goodpasture syndrome, Degos disease, and fulminant antiphospholipid syndrome. (Item 26) The complement-related condition is atypical hemolytic uremic syndrome (aHUS), as described in item 24. (Item 27) The complement-related condition is paroxysmal nocturnal hemoglobinuria (PNH), as described in item 24. (Item 28) The method according to any one of items 24 to 26, wherein the stable aqueous solution is administered subcutaneously to the patient. (Item 29) The method according to any one of items 24 to 26, wherein the stable aqueous solution is administered intravenously to the patient. (Item 30) A therapeutic kit comprising (i) the stable aqueous solution described in any one of items 1 to 21, and (ii) means for delivering the stable aqueous solution to a human. (Item 31) The therapeutic kit described in item 28, wherein the means is a syringe. [Brief explanation of the drawing]

[0059] [Figure 1] The results of the dynamic light scattering method for the salt titration of ravulizumab (ALXN1210) in histidine buffer at 50 mg / mL are shown. [Figure 2] The results of the dynamic light scattering method for the L-arginine titration of 50 mg / mL buffer-exchanged ravulizumab (ALXN1210) are shown. [Figure 3] The results of the dynamic light scattering method for the salt titration of 50 mg / mL phosphate-buffered ravulizumab (ALXN1210) are shown. [Figure 4] The results of differential scanning fluorescence imaging for 50 mg / mL buffer-exchanged ravulizumab (ALXN1210) are shown. [Figure 5] The results of dynamic light scattering for ravulizumab (ALXN1210) at concentrations of 10 mg / mL and 114 mg / mL without L-arginine, and for ravulizumab (ALXN1210) at concentration of 114 mg / mL with added L-arginine are shown. [Figure 6] The stability data for ravulizumab (ALXN1210) is shown (T=0 to T=2 weeks, 2 to 8°C). [Figure 7] The stability data for ravulizumab (ALXN1210) is shown (T=3 weeks to T=2 months, 2-8°C). [Figure 8] The stability data for ravulizumab (ALXN1210) is shown (T=0 to T=3 weeks, 23 to 27°C). [Figure 9] The stability data for ravulizumab (ALXN1210) is shown (T=January to T=February, 23-27°C). [Figure 10] The stability data for ravulizumab (ALXN1210) is shown (T=1 week to T=3 weeks, 37°C). [Figure 11] The stability data for ravulizumab (ALXN1210) is shown (T=January to T=February, 37°C). [Figure 12] Stability size exclusion chromatography (SEC) data for ravulizumab (ALXN1210), showing monomer percentage (T=0 to T=2 months, 2 to 8°C). [Figure 13] Stability size exclusion chromatography (SEC) data for ravulizumab (ALXN1210), showing monomer percentage (T=0 to T=2 months, 23-27°C). [Figure 14] Stability size exclusion chromatography (SEC) data for ravulizumab (ALXN1210), showing monomer percentage (T=0 to T=2 months, 37°C). [Figure 15] Stability dynamic light scattering data for ravulizumab (ALXN1210) and histidine samples (T=0) are shown. [Figure 16] The data for the stability of the ravulizumab (ALXN1210) histidine AS sample (T=0) is shown using dynamic light scattering. [Figure 17] The data for the stability of the ravulizumab (ALXN1210) phosphate sample (T=0) is shown using dynamic light scattering. [Figure 18] The data for the stability of ravulizumab (ALXN1210) phosphate samples (T=February, 2-8°C) are shown using dynamic light scattering. [Figure 19] The freeze-thaw stability data for ravulizumab (ALXN1210) is shown (T=0 to cycle 2, T=1M at -20℃). [Figure 20] The stability data for ravulizumab (ALXN1210) under freeze-thaw cycles 3 to 5, T=1M, and -20°C is shown. [Figure 21] Stability size exclusion chromatography (SEC) data for ravulizumab (ALXN1210), showing monomer percentage (T=1 month, freeze-thaw cycle at -20°C). [Figure 22] Stability data for the prototype of ravulizumab (ALXN1210) is shown (T=0 months to T=2 months, 2 to 8°C). [Figure 23] Stability data for the prototype of ravulizumab (ALXN1210) is shown (T=March to T=June, 2-8°C). [Figure 24] This shows the stability data for the prototype of ravulizumab (ALXN1210) (T=September to T=December, 2-8°C). [Figure 25] Stability data for the prototype of ravulizumab (ALXN1210) is shown (T=January to T=February, 23-27°C). [Figure 26] Stability data for the prototype of ravulizumab (ALXN1210) is shown (T=March to T=June, 23-27°C). [Figure 27] This shows the stability data for the prototype of ravulizumab (ALXN1210) (T=September to T=December, 23-27°C). [Figure 28] Stability data for the prototype of ravulizumab (ALXN1210) is shown (T=2 weeks to T=2 months, 37°C). [Figure 29] Stability data for the prototype of ravulizumab (ALXN1210) is shown (T=January to T=March, -20°C). [Figure 30] Stability data for the prototype of ravulizumab (ALXN1210) is shown (T=June to T=December, -20°C). [Figure 31] The results of the stability test of the ravulizumab (ALXN1210) prototype are shown (T=March to T=June, -80°C). [Figure 32] The results of the stability test of the ravulizumab (ALXN1210) prototype are shown (T=December, -80°C). [Figure 33] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing the monomer percentage (T=0 to T=12 months, 2 to 8°C). [Figure 34] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing monomer percentage (T=0 to T=12 months, 23-27°C). [Figure 35]Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing monomer percentage (T=0 to T=12 months, 37°C). [Figure 36] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing the monomer percentage (T=0 to T=12 months, -20°C). [Figure 37] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing monomer percentage (T=0 to T=12 months, -80°C). [Figure 38] Stability data from dynamic light scattering of a prototype 75 mg / mL ravulizumab ALXN1210 phosphate sample (T=0) are shown. [Figure 39] Stability data from dynamic light scattering of a 75 mg / mL ravulizumab ALXN1210 phosphate sample (T=1 month, 2-8°C) are shown. [Figure 40] Stability data from dynamic light scattering of a 100 mg / mL ravulizumab ALXN1210 phosphate sample (T=1 month, 2-8°C) are shown. [Figure 41] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing monomer percentage - freeze-thaw cycles 1-3, T=1 month, -20°C. [Figure 42] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing monomer percentage - freeze-thaw cycles 4-5, T=1 month, -20°C. [Figure 43] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing monomer percentage - freeze-thaw cycles 1-3, T=3 months, -80°C. [Figure 44] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing monomer percentage - freeze-thaw cycles 4-5, T=3 months, -80°C. [Figure 45] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing monomer percentage. Freeze-thaw cycles 1-5, T=1 month, -20°C. [Figure 46] Stability size exclusion chromatography (SEC) data for the prototype of ravulizumab (ALXN1210), showing monomer percentage. Freeze-thaw cycles 1-5, T=3 months, -80°C. [Figure 47] This describes the overall design of a Phase I trial designed to evaluate the safety, tolerability, pharmacokinetics (PK), disease progression (PD), and immunogenicity of a single 400 mg subcutaneous dose of ravulizumab (ALXN1210) compared to a single 400 mg intravenously administered dose of ravulizumab (ALXN1210) or a subcutaneously administered placebo in 42 healthy subjects. [Figure 48] A summary of the ongoing status of all clinical trials will be presented. [Figure 49] This graph shows individual ALXN1210 serum concentrations relative to the scheduled time, using a linear scale. [Figure 50] This graph shows individual ALXN1210 serum concentrations relative to the scheduled time, using a log-linear scale. [Figure 51] This graph shows the mean (±SD) percentage change in free C5 serum concentration over time from baseline for subjects who received placebo SC, ALXN1210 SC, and ALXN1210 IV. [Figure 52] This graph shows the mean (±SD) percentage change in total C5 serum concentration over time from baseline for subjects who received placebo SC, ALXN1210 SC, and ALXN1210 IV. [Figure 53] This graph shows the mean (±SD) percentage change in chicken erythrocyte (cRBC) hemolysis over time from baseline in subjects who received placebo SC, ALXN1210 SC, and ALXN1210 IV. [Modes for carrying out the invention]

[0060] This disclosure features a stable aqueous solution containing a high concentration of an anti-C5 antibody (e.g., ravulizumab). The solution can be used in a variety of therapeutic applications, such as the treatment or prevention of complement-related disorders. Without intent to limit, exemplary solutions, formulations, therapeutic kits, and methods for preparing and using any of the foregoing are described in detail below and illustrated in the examples.

[0061] I. definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those universally understood by those skilled in the art. Any methods and compositions similar to or equivalent to those described herein may be used in carrying out or verifying the present invention, but preferred methods and compositions are described herein.

[0062] The singular forms "a," "an," and "the" refer to multiple objects unless otherwise explicitly indicated by the context.

[0063] The term "approximately" means to include a deviation within plus or minus 10 percent (±10%), especially when relating to a given quantity or number (e.g., ±5%).

[0064] The term "pharmaceutical preparation" refers to a preparation in which the biological activity of an active ingredient is clearly effective, and which does not contain any additional components that are significantly toxic to the subject to which the preparation is administered.

[0065] As used herein, an "aqueous" pharmaceutical composition is a composition suitable for pharmaceutical use, in which case the aqueous carrier is water. A composition suitable for pharmaceutical use may be sterile, homogeneous, and / or isotonic. Aqueous pharmaceutical compositions may be prepared directly in aqueous form and / or reconstituted from lyophilized products.

[0066] An "isotonic" preparation is one that has essentially the same osmotic pressure as human blood. Isotonic preparations generally have an osmotic pressure of approximately 275–350 mOsm / kg. The term "hypotonic" describes a preparation that has an osmotic pressure lower than that of human blood. Accordingly, the term "hypertonic" is used to describe a preparation that has an osmotic pressure higher than that of human blood. Isotonicity can be measured, for example, using vapor pressure or an ice-freezing osmometer. An "isotonic agent" is a compound that makes a preparation isotonic.

[0067] As used herein, the “osmotic pressure” of a solution is the number of osmoles of solute per kilogram of solvent. Osmotic pressure is a unit of measurement for the number of particles present in a solution and is independent of the size or weight of the particles. It can only be measured by using properties of the solution that depend solely on the particle concentration. These properties are vapor pressure depression, freezing point depression, boiling point elevation, and osmotic pressure, and are collectively referred to as colligative properties.

[0068] A "sterilized" preparation is either sterile, or free from or substantially free from all living microorganisms or their spores.

[0069] A “stable” formulation, as used herein, is a formulation in which the antibody in the formulation inherently retains its physical and / or chemical stability and / or biological activity during storage. Various analytical techniques for measuring protein stability are available in the art and are summarized in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). The stability of anti-C5 antibody formulations can be measured at a selected temperature after a selected period. For example, increased agglutination after storage indicates instability of aqueous anti-C5 antibody formulations. In addition to agglutination, retention of the original clarity, color, and odor throughout the shelf life are indicators used to monitor the stability of anti-C5 antibody aqueous solutions described herein.

[0070] If, upon visual inspection of color and / or clarity, or when measured by UV light scattering or size exclusion chromatography, there are virtually no signs of aggregation, precipitation, and / or denaturation, the antibody is considered to have "maintained its physical stability" in the pharmaceutical formulation.

[0071] The term "aggregation" refers to the assembly of naturally folded proteins into aggregates containing non-natural structures. Aggregation can occur even under physiologically non-denaturing conditions, and is often irreversible, resulting in non-natural aggregates that are inert, and sometimes immunogenic and toxic.

[0072] As used herein, the term "low to undetectable levels of aggregation" refers to samples containing aggregation of approximately 5% or less, approximately 4% or less, approximately 3% or less, approximately 2% or less, approximately 1% or less, and approximately 0.5% or less of the protein weight, as measured by gel permeation high-performance liquid chromatography (GP-HPLC), high-speed size exclusion chromatography (HPSEC), or static light scattering (SLS).

[0073] Chemical stability at a given point in time is defined as the ability of an antibody to retain its biological activity, in which case the antibody "retains its chemical stability" in a pharmaceutical formulation. Chemical stability can be assessed by detecting and quantifying chemically altered forms of the antibody. Chemical alterations may include size modification (e.g., clipping), deamidation, racemization, hydrolysis, oxidation, beta-elimination, and disulfide exchange, which can be assessed by known techniques, such as size exclusion chromatography, SDS-PAGE, matrix-assisted laser desorption / ionization / time-of-flight mass spectrometry (MALDI / TOF MS), and / or ion-exchange chromatography.

[0074] An antibody “retains its biological activity” in a pharmaceutical formulation if it is biologically active for its intended purpose within the formulation. For example, biological activity is retained if the biological activity of the antibody in the pharmaceutical formulation is within approximately 30%, 20%, or 10% (within the assay error range) of the biological activity exhibited at the time the pharmaceutical formulation was prepared (determined, e.g., by an antigen-binding assay). In this specification, “biological activity” of a monoclonal antibody refers to the antibody’s ability to bind to an antigen. Further, antibodies may include those that bind to an antigen and thereby produce a measurable biological response that can be measured in vitro or in vivo.

[0075] The "shelf life" of a pharmaceutical product, such as an aqueous solution containing an anti-C5 antibody, is the length of time the product can be stored before decomposition occurs. For example, the shelf life may be defined as the time it takes for 0.1%, 0.5%, 1%, 5%, or 10% of the product to decompose.

[0076] As used herein, the term "antibody" describes a polypeptide that includes an antigen-binding site (e.g., VH / VL region or Fv or CDR) derived from at least one antibody. Antibodies include known antibody types. For example, an antibody may be a human antibody, a humanized antibody, a bispecific antibody, or a chimeric antibody. An antibody may be a Fab, Fab’2, ScFv, SMIP, Affibody®, nanobody or domain antibody. An antibody may further be any of the following isotypes: IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgAsec, IgD and IgE. An antibody may be a natural antibody or an antibody modified by protein modification techniques (e.g., mutation, deletion, substitution, complexation with non-antibody moieties, etc.). For example, an antibody may include one or more variant amino acids that change the properties (e.g., functional properties) of the antibody (compared to the native antibody). For example, many such modifications that affect half-life, effector function, and / or the immune response to the antibody in a patient are known in the art. The term "antibody" also includes artificial or modified polypeptide constructs that contain an antigen-binding site derived from at least one antibody.

[0077] As used herein, the terms "specific binding", "selective binding", "selectively binds" and "specifically binds" refer to an antibody that binds to an epitope on a defined antigen but does not bind to other antigens. Typically, an antibody has an equilibrium dissociation constant (K -7 less than 10 -8 M, e.g., approximately 10 -9 M, less than 10 -10 M or less or even lower, determined by surface plasmon resonance (SPR) technology, e.g., in a BIACORE® 2000 surface plasmon resonance apparatus, using a defined antigen such as C5 as an analyte and an antibody as a ligand, or by Scatchard analysis of the antibody against antigen-positive cells. D(ii) binds to the given antigen with an affinity at least twice as high as the affinity to nonspecific antigens other than the given antigen or closely related antigens (e.g., BSA, casein, etc.). Therefore, unless otherwise indicated, an antibody that "specifically binds to human C5" means 10 -7 M or less, for example, approximately 10 -8 Less than M, 10 -9 Less than M or 10 -10 K less than M or even lower D This refers to antibodies that bind to soluble or cell-bound human C5.

[0078] As used herein, the term "surface plasmon resonance" refers to an optical phenomenon that enables real-time analysis of biospecific interactions by detecting changes in protein concentration within a biosensor matrix using, for example, the BIAcore system (Pharmacia Biosensor AB, Uppsala, Sweden, and Piscataway, New Jersey). For further details, see Jonsson, U., et al. (1993) Ann. Biol. Clin. 51:19-26; Jonsson, U., et al. (1991) Biotechniques 11:620-627; Johnsson, B., et al. (1995) J. Mol. Recognit. 8:125-131; and Johnsson, B., et al. (1991) Anal. Biochem. 198:268-277.

[0079] When used herein, "K off The term "dissociation rate constant" is intended to refer to the dissociation rate constant at which an antibody dissociates from an antibody / antigen complex.

[0080] When used herein, "K d The term "dissociation constant" is intended to refer to the dissociation constant of a particular antibody-antigen interaction.

[0081] As used herein, the terms “subject” or “patient” are interchangeable herein and refer to mammals such as humans, mice, rats, hamsters, guinea pigs, rabbits, cats, dogs, monkeys, cattle, horses, and pigs. In one embodiment, the patient is a human patient (e.g., a human patient having a complement-related condition).

[0082] As used herein, the terms “to treat,” “to treat,” and “treatment” refer to the therapeutic means described herein. A “treatment” method involves administering to a subject any combination of the treatments disclosed herein in order to cure, delay, reduce the severity of, or improve one or more symptoms of a disease or disorder or a recurrent disease or disorder, or to extend the subject’s survival beyond the period that would be expected if such treatment were not performed.

[0083] As used herein, “effective treatment” means treatment that produces a beneficial effect, such as improvement of at least one symptom of a disease or disorder. Beneficial effects may also take the form of improvement exceeding a standard, i.e., improvement exceeding the results of measurements or observations made before initiating treatment according to this method. Effective treatment may also mean the alleviation of at least one symptom of a disease or condition.

[0084] The term “effective dose” refers to the amount of an agent that produces a desired biological, therapeutic, and / or preventive outcome. This outcome may be a reduction, improvement, relief, attenuation, delay, and / or mitigation of one or more signs, symptoms, or causes of a disease or condition, or any other desirable change in the biological system. In one example, an “effective dose” is the amount of a stable aqueous solution that alleviates at least one symptom of a disease or condition. An effective dose may be administered in one or more doses.

[0085] As used herein, the terms “induction” and “induction phase” are interchangeable and refer to the first phase of treatment.

[0086] As used herein, the terms “maintenance” and “maintenance phase” are interchangeable and refer to the second phase of treatment. In certain embodiments, treatment is continued as long as a clinical benefit is observed or until uncontrollable toxicity or disease progression occurs.

[0087] II. Anti-C5 antibody The anti-C5 antibodies described herein bind to complement component C5 (e.g., human C5) and inhibit its cleavage into C5a and C5b fragments. As described above, these antibodies also exhibit improved pharmacokinetic properties compared to other anti-C5 antibodies used for therapeutic purposes (e.g., eculizumab).

[0088] Anti-C5 antibodies (or VH / VL domains derived therefrom) suitable for use in the present invention can be produced using methods well known in the art. Alternatively, anti-C5 antibodies recognized in the art can be used. Antibodies that bind to C5 and compete with any of these recognized antibodies in the art can also be used.

[0089] Examples of anti-C5 antibodies include ravulizumab, or its antigen-binding fragments and variants thereof, which contain heavy and light chains having the sequences shown in SEQ ID NOs. 14 and 11, respectively. Rabulizumab (also known as BNJ441 and ALXN1210) is described in International Patent Application No. PCT / US2015 / 019225 and U.S. Patent No. 9,079,949, the teachings of which are incorporated herein by reference. The terms ravulizumab, BNJ441, and ALXN1210 may be used interchangeably throughout this document. Rabulizumab selectively binds to human complement protein C5 and inhibits the cleavage of C5 into C5a and C5b during complement activation. This inhibition disrupts the release of the pro-inflammatory mediator C5a and the formation of C5b-9 of the membrane invasion complex (MAC), which forms cytolytic pores, while conserving the basal or early components of complement activation (e.g., C3 and C3b), which are essential for microbial opsonization and immune complex clearance.

[0090] In other embodiments, the antibody comprises the CDR or variable region of the heavy and light chains of ravulizumab. Thus, in one embodiment, the antibody comprises the CDR1, CDR2, and CDR3 domains of the VH region of ravulizumab having the sequence described in SEQ ID NO: 12, and the CDR1, CDR2, and CDR3 domains of the VL region of ravulizumab having the sequence described in SEQ ID NO: 8. In another embodiment, the antibody comprises the heavy chain CDR1, CDR2, and CDR3 domains having the sequences described in SEQ ID NOs: 19, 18, and 3, respectively, and the light chain CDR1, CDR2, and CDR3 domains having the sequences described in SEQ ID NOs: 4, 5, and 6, respectively. In yet another embodiment, the antibody comprises the VH region and the VL region having the amino acid sequences described in SEQ ID NO: 12 and SEQ ID NO: 8, respectively.

[0091] Another exemplary anti-C5 antibody is antibody BNJ421, which comprises a heavy chain and a light chain having the sequences shown in SEQ ID NOs. 20 and 11, respectively, or its antigen-binding fragment and variants. BNJ421 (also known as ALXN1211) is described in International Patent Application No. PCT / US2015 / 019225 and U.S. Patent No. 9,079,949, the teachings of which are incorporated herein by reference.

[0092] In other embodiments, the antibody comprises the CDR or variable region of the heavy and light chains of BNJ421. Thus, in one embodiment, the antibody comprises the CDR1, CDR2, and CDR3 domains of the VH region of BNJ421 having the sequence described in SEQ ID NO: 12, and the CDR1, CDR2, and CDR3 domains of the VL region of BNJ421 having the sequence described in SEQ ID NO: 8. In another embodiment, the antibody comprises the heavy chain CDR1, CDR2, and CDR3 domains having the sequences described in SEQ ID NOs: 19, 18, and 3, respectively, and the light chain CDR1, CDR2, and CDR3 domains having the sequences described in SEQ ID NOs: 4, 5, and 6, respectively. In yet another embodiment, the antibody comprises the VH region and the VL region having the amino acid sequences described in SEQ ID NO: 12 and SEQ ID NO: 8, respectively.

[0093] The precise boundaries of the CDR are defined in various ways according to different methods. In some embodiments, the location of the CDR or framework region within the light chain variable domain or heavy chain variable domain is described in Kabat et al.'s [(1991) “Sequences of Proteins”] The CDR may be as defined in “of Immunological Interest.” NIH Publication No. 91-3242, USD Department of Health and Human Services, Bethesda, MD. In such cases, the CDR may be referred to as “Kabat CDR” (e.g., “Kabat LCDR2” or “Kabat HCDR1”). In some embodiments, the location of the CDR in the light chain variable region or heavy chain variable region is as defined in Chothia et al. (1989) Nature 342 These can be as defined in :877-883. Therefore, these regions may be referred to as "Chothia CDR" (e.g., "Chothia LCDR2" or "Chothia HCDR3"). In some embodiments, the locations of the light-chain variable region and the heavy-chain variable region of the CDR can be as defined by the combined definition of Kabat and Chothia (Kabat-Chothia). In such embodiments, these regions may be referred to as "Kabat-Chothia combination CDR". Thomas et al. [(1996) Mol Immunol 33(17 / 18) [1389-1401] illustrates the identification of CDR boundaries according to the definitions of Kabat and Chothia.

[0094] In some embodiments, the anti-C5 antibody described herein contains a heavy chain CDR1 comprising or consisting of the following amino acid sequence:G H IFSNYWIQ (SEQ ID NO: 19). In some embodiments, the anti-C5 antibody described herein comprises a heavy chain CDR2 having or consisting of the following amino acid sequence: EILPGSGH TEYTENFKD (SEQ ID NO: 18). In some embodiments, the anti-C5 antibodies described herein include a heavy chain variable region comprising the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASG H IFSNYWIQWVRQAPGQGLEWMGEILPGSG H TEYTENFKDRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARYFFGSSPNWYFDVWGQGTLVTVSS(Sequence ID 12).

[0095] In some embodiments, the anti-C5 antibody described herein includes a light chain variable region comprising the following amino acid sequence:DIQMTQSPSSLSASVGDRVTITCGASENIYGALNWYQQKPGKAPKLLIYGATNLADGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQNVLNTPLTFGQGTKVEIK (SEQ ID NO: 8).

[0096] In some embodiments, the anti-C5 antibodies described herein may contain a mutant human Fc constant region that binds to a human neonatal Fc receptor (FcRn) with a higher affinity than the native human Fc constant region from which the mutant human Fc constant region is induced. For example, the Fc constant region may contain one or more (e.g., 2, 3, 4, 5, 6, 7, or 8 or more) amino acid substitutions compared to the native human Fc constant region from which the mutant human Fc constant region is induced. The substitutions can increase the binding affinity of an IgG antibody containing the mutant Fc constant region to FcRn at pH 6.0 while maintaining the pH dependence of the interaction. Methods for testing whether one or more substitutions in the Fc constant region of an antibody increase the affinity of the Fc constant region to FcRn at pH 6.0 (while maintaining the pH dependence of the interaction) are known in the art and are illustrated in the examples. See, for example, International Patent Application No. PCT / US2015 / 019225 and U.S. Patent No. 9,079949, the respective disclosures of which are incorporated herein by reference in their entirety.

[0097] Substitutions that enhance the binding affinity of the antibody Fc constant region to FcRn are known in this field, for example, (1) Dall'Acqua et al. (2006) J Biol Chem 281 (2) Triple substitution of M252Y / S254T / T256E as described in 23514-23524, (2) Hinton et al. (2004) J Biol Chem 279 :6213-6216 and Hinton et al. (2006) J Immunol 176 Substitutions of M428L or T250Q / M428L as described in 346-356, and (3) Petkova et al. (2006) Int Immunol 18(12) Substitutions include N434A or T307 / E380A / N434A as described in 1759-69. Further substitution combinations include P257I / Q311I, P257I / N434H, and D376V / N434H, for example, Datta-Mannan et al. (2007) J Biol. Chem 282(3) This is described in 1709-1717, and its disclosure is incorporated herein by reference in its entirety.

[0098] In some embodiments, the variant constant region has a substitution for valine at EU amino acid residue 255. In some embodiments, the variant constant region has a substitution for asparagine at EU amino acid residue 309. In some embodiments, the variant constant region has a substitution for isoleucine at EU amino acid residue 312. In some embodiments, the variant constant region has a substitution at EU amino acid residue 386.

[0099] In some embodiments, the mutant Fc constant region includes 30 or fewer amino acid substitutions, insertions, or deletions (e.g., 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2) compared to the native constant region from which it originates. In some embodiments, the mutant Fc constant region includes one or more amino acid substitutions selected from the group consisting of: M252Y, S254T, T256E, N434S, M428L, V259I, T250I, and V308F. In some embodiments, the mutant human Fc constant region contains methionine at position 428 and asparagine at position 434, respectively, according to EU numbering. In some embodiments, the mutant Fc constant region contains two substitutions, for example, 428L / 434S, as described in U.S. Patent No. 8,088,376.

[0100] In some embodiments, the precise locations of these mutations may be shifted from the native human Fc constant region locations by antibody engineering. For example, when used in IgG2 / 4 chimeric Fc, the two substitutions 428L / 434S may correspond to 429L and 435S, as seen in the M429L and N435S variants found in ravulizumab (BNJ441) and described in U.S. Patent No. 9,079,949, which is incorporated herein by reference in its entirety.

[0101] In some embodiments, the mutant constant region includes substitutions at amino acid positions 237, 238, 239, 248, 250, 252, 254, 255, 256, 257, 258, 265, 270, 286, 289, 297, 298, 303, 305, 307, 308, 309, 311, 312, 314, 315, 317, 325, 332, 334, 360, 376, 380, 382, ​​384, 385, 386, 387, 389, 424, 428, 433, 434, or 436 (EU numbering) compared to the native human Fc constant region. In some embodiments, substitutions include: methionine for glycine at position 237; alanine for proline at position 238; lysine for serine at position 239; isoleucine for lysine at position 248; alanine, phenylalanine, isoleucine, methionine, glutamine, serine, valine, tryptophan, or tyrosine for threonine at position 250; phenylalanine, tryptophan, or tyrosine for methionine at position 252; threonine for serine at position 254; glutamic acid for arginine at position 255; and aspartic acid or glutamic acid for threonine at position 256. , or glutamine; alanine, glycine, isoleucine, leucine, methionine, asparagine, serine, threonine, or valine relative to proline at position 257; histidine relative to glutamic acid at position 258; alanine relative to aspartic acid at position 265; phenylalanine relative to aspartic acid at position 270; alanine or glutamic acid relative to asparagine at position 286; histidine relative to threonine at position 289; alanine relative to asparagine at position 297; glycine relative to serine at position 298; alanine relative to valine at position 303; alanine relative to valine at position 305;Alanine, aspartic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, valine, tryptophan, or tyrosine relative to threonine at position 307; alanine, phenylalanine, isoleucine, leucine, methionine, proline, glutamine, or threonine relative to valine at position 308; alanine, aspartic acid, glutamic acid, proline, or arginine relative to leucine or valine at position 309. Arginine; alanine, histidine, or isoleucine for glutamine at position 311; alanine or histidine for aspartic acid at position 312; lysine or arginine for leucine at position 314; alanine or histidine for asparagine at position 315; alanine for lysine at position 317; glycine for asparagine at position 325; valine for isoleucine at position 332; leucine for lysine at position 334; histidine for lysine at position 360 Alanine for aspartic acid at position 376; alanine for glutamic acid at position 380; alanine for glutamic acid at position 382; alanine for asparagine or serine at position 384; aspartic acid or histidine for glycine at position 385; proline for glutamine at position 386; glutamic acid for proline at position 387; alanine or serine for asparagine at position 389; alanine for serine at position 424; and methionine at position 428. The following are selected from the group consisting of alanine, aspartic acid, phenylalanine, glycine, histidine, isoleucine, lysine, asparagine, proline, glutamine, serine, threonine, valine, tryptophan, or tyrosine; lysine instead of histidine at position 433; alanine, phenylalanine, histidine, serine, tryptophan, or tyrosine instead of asparagine at position 434; and histidine instead of tyrosine or phenylalanine at position 436, all according to EU numbering.

[0102] In some embodiments, an anti-C5 antibody suitable for use in the method described herein comprises a heavy chain polypeptide containing the amino acid sequence shown in SEQ ID NO: 14 and / or a light chain polypeptide containing the amino acid sequence shown in SEQ ID NO: 11. Alternatively, in some embodiments, an anti-C5 antibody suitable for use in the method described herein comprises a heavy chain polypeptide containing the amino acid sequence shown in SEQ ID NO: 20 and / or a light chain polypeptide containing the amino acid sequence shown in SEQ ID NO: 11.

[0103] In one embodiment, the antibody has an affinity dissociation constant (K) of at least 0.1 (e.g., at least 0.15, 0.175, 0.2, 0.25, 0.275, 0.3, 0.325, 0.35, 0.375, 0.4, 0.425, 0.45, 0.475, 0.5, 0.525, 0.55, 0.575, 0.6, 0.625, 0.65, 0.675, 0.7, 0.725, 0.75, 0.775, 0.8, 0.825, 0.85, 0.875, 0.9, 0.925, 0.95 or 0.975) nM. D ) binds to C5 at pH 7.4 and 25°C (or under physiological conditions by other means). In some embodiments, the anti-C5 antibody or its antigen-binding fragment K D This is less than or equal to 1 nM (for example, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, or 0.2 nM or less).

[0104] In other embodiments, [(pH 6.0, °C, K of antibody against C5] D ) / (K of antibody against C5 at pH 7.4, 25℃) D)] is greater than 21 (for example, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, (Greater than 5000, 5500, 6000, 6500, 7000, 7500, or 8000).

[0105] Methods for determining whether an antibody is bound to a protein antigen and / or the affinity of an antibody to a protein antigen are known in the art. For example, antibody binding to protein antigens can be determined by Western blotting, dot blotting, surface plasmon resonance (SPR) (e.g., BIAcore system; Pharmacia Biosensor). It can be detected and / or quantified using various techniques such as enzyme-linked immunosorbent assay (ELISA) or AB (Agency for Antibody Engineering, Sweden, Uppsala and Piscataway, New Jersey). For example, Benny KCLo (2004) “Antibody Engineering: Methods and Protocols,” Humana Press (ISBN: 1588290921); Johne et al. (1993) J Immunol Meth 160 :191-198;Jonsson et al.(1993)Ann Biol Clin 51 :19-26; and Jonsson et al. (1991) Biotechniques 11 See 620-627. Furthermore, methods for measuring affinity (e.g., dissociation and association constants) are described in the examples.

[0106] When used herein, "K a The term "k" refers to the rate constant at which antibodies associate with an antigen. dThe term "K" refers to the rate constant for the dissociation of an antibody from an antibody / antigen complex. D The term "equilibrium dissociation constant" refers to the equilibrium dissociation constant of antibody-antigen interactions. The equilibrium dissociation constant is the velocity constant K D = k a / k d This is estimated from the ratio. Such determinations are preferably measured at 25°C or 37°C (see Examples). For example, the kinetics of antibodies binding to human C5 can be determined at pH 8.0, 7.4, 7.0, 6.5, and 6.0 via surface plasmon resonance (SPR) on a BIAcore 3000 instrument using an anti-Fc antibody capture method to immobilize the antibody.

[0107] In one embodiment, an anti-C5 antibody or its antigen-binding fragment inhibits the generation or activity of the C5a and / or C5b active fragments of the C5 protein (e.g., human C5 protein). Through this inhibitory action, the antibody inhibits, for example, the pro-inflammatory effects of C5a and the formation of the C5b-9 membrane invasion complex (MAC) on the cell surface.

[0108] Methods for determining whether specific antibodies described herein inhibit C5 cleavage are known in the art. Inhibition of human complement component C5 can reduce the cytolytic capacity of complement in the target body fluid. Such a reduction in the cytolytic capacity of complement present in body fluid is described, for example, in Kabat and Mayer (eds), "Experimental Immunochemistry, 2 nd Conventional hemolytic assays, such as the hemolytic assay described in "Edition," 135-240, Springfield, IL, CC Thomas (1961), pages 135-139, or, for example, Hillmen et al. (2004) N Engl J Med 350(6)Measurement can be performed by various conventional assays, such as the hemolysis method for chicken red blood cells described in :552, which are well known in the art. Methods for determining whether a candidate compound inhibits the cleavage of human C5 to morphologies C5a and C5b are well known in the art, for example, Evans et al. (1995) Mol Immunol 32(16) As described in 1183-95, for example, the concentrations and / or physiological activities of C5a and C5b in body fluids can be measured by methods well known in the art. For C5b, hemolytic assays or assays for soluble C5b-9 described herein can be used. Other assays known in the art can also be used. These assays, or other suitable assay types, can be used to screen for candidate substances that can inhibit human complement component C5.

[0109] For example, immunoassay techniques such as ELISA can be used to measure the protein concentrations of C5 and / or its fragmentation products, and to determine the ability of an anti-C5 antibody or its antigen-binding fragment to inhibit the conversion of C5 to a bioactive product. In some embodiments, C5a production is measured. In some embodiments, C5b-9 neoepitope-specific antibodies are used to detect terminal complement formation.

[0110] A hemolysis assay can be used to determine the inhibitory activity of an anti-C5 antibody or its antigen-binding fragment against complement activation. To determine the effect of an anti-C5 antibody or its antigen-binding fragment against classical complement pathway-mediated hemolysis in an in vitro serum test solution, hemolysin-coated sheep erythrocytes or chicken erythrocytes sensitive with an anti-chicken erythrocyte antibody are used as target cells. The percentage of hemolysis is standardized by setting the lysis occurring in the absence of the inhibitor to 100%. In some embodiments, for example, as used in the Wieslab® Classical Pathway Complement Kit (Wieslab® COMPL CP310, Euro-Diagnostica, Sweden), the classical complement pathway is activated by a human IgM antibody. Briefly, the test serum is incubated with the anti-C5 antibody or its antigen-binding fragment in the presence of a human IgM antibody. The amount of C5b-9 produced is measured by contacting the enzyme, which is complexed with an anti-C5b-9 antibody, and a fluorescence-generating substrate with this mixture and measuring the absorbance at an appropriate wavelength. As a control, the test serum is incubated in the absence of an anti-C5 antibody or its antigen-binding fragment. In some embodiments, the test serum is C5-deficient serum reconstituted with a C5 polypeptide.

[0111] To determine the effect of anti-C5 antibodies or their antigen-binding fragments on alternative pathway-mediated hemolysis, unsustainable rabbit or guinea pig erythrocytes can be used as target cells. In some embodiments, the serum test solution is C5-deficient serum reconstituted with a C5 polypeptide. The hemolysis rate is standardized by setting the lysis occurring in the absence of the inhibitor to 100%. In some embodiments, for example, as used in the Wieslab® Alternative Pathway Complement Kit (Wieslab® COMPL AP330, Euro-Diagnostica, Sweden), the alternative complement pathway is activated by a lipopolysaccharide molecule. Briefly, the test serum is incubated with an anti-C5 antibody or its antigen-binding fragment in the presence of a lipopolysaccharide. The amount of C5b-9 produced is measured by contacting this mixture with an enzyme complexed with the anti-C5b-9 antibody and a fluorescent substrate, and measuring the fluorescence at an appropriate wavelength. As a control, the test serum is incubated in the absence of anti-C5 antibody or its antigen-binding fragment.

[0112] In some embodiments, C5 activity or its inhibition is quantified using the CH50eq assay. The CH50eq assay is a method for measuring total classical complement activity in serum. This test is a lysis assay that uses antibody-sensitive erythrocytes as activators of the classical complement pathway and test serum of various dilutions to determine the amount required to obtain 50% lysis (CH50). The percentage of hemolysis can be determined, for example, using a spectrophotometer. The CH50eq assay provides an indirect measure of terminal complement complex (TCC) formation, the TCC itself being the direct cause of the hemolysis being measured.

[0113] This assay is well known and has been universally performed by those skilled in the art. Briefly, an undiluted serum sample (e.g., a reconstituted human serum sample) is added to a microassay well containing antibody-sensitive erythrocytes to activate the classical complement pathway and generate triglycerides (TCCs). The activated serum is then diluted in microassay wells coated with a capture reagent (e.g., an antibody that binds to one or more components of TCCs). The TCCs present in the activated sample bind to the monoclonal antibody covering the surface of the microassay wells. The wells are washed, and a detection reagent that recognizes the bound TCCs, which has been labeled for detection, is added to each well. The detection label may be, for example, fluorescent or enzymatic. The assay results are expressed in units of CH50 equivalents per milliliter (CH50 U Eq / mL).

[0114] For example, inhibition associated with terminal complement activity includes a reduction of at least 5% (e.g., at least 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60%) in terminal complement activity, for example in a hemolysis assay or CH50eq assay, compared to the effect of a control antibody (or its antigen-binding fragment) under similar conditions and at equimolar concentrations. As used herein, substantial inhibition refers to inhibition of a given activity (e.g., terminal complement activity) of at least 40% (e.g., at least 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95%). In some embodiments, the anti-C5 antibodies described herein contain one or more amino acid substitutions compared to the CDR of eculizumab (i.e., SEQ ID NOs: 1-6), but retain at least 30% (e.g., at least 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95%) of the complement inhibitory activity of eculizumab in a hemolysis assay or CH50eq assay.

[0115] The anti-C5 antibodies described herein have a serum half-life in humans of at least 20 days (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 days). In another embodiment, the anti-C5 antibodies described herein have a serum half-life of at least 40 days in humans. In another embodiment, the anti-C5 antibodies described herein have a serum half-life of approximately 43 days in humans. In another embodiment, the anti-C5 antibodies described herein have a serum half-life of 39 to 48 days in humans. Methods for measuring the serum half-life of antibodies are known in the art. In some embodiments, the anti-C5 antibodies or antigen-binding fragments described herein have a serum half-life at least 20% (e.g., at least 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 250, 300, 400, 500%) higher than the serum half-life of eculizumab when measured in one of the mouse model systems described in the examples (e.g., C5-deficient / NOD / SCID mouse model system or hFcRn transgenic mouse model system).

[0116] In one embodiment, an antibody competes for binding to the same C5 epitope as the antibody described herein, and / or binds to the same C5 epitope as the antibody described herein. The phrase “binding to the same epitope” when referring to two or more antibodies means that the antibodies bind to the same segment of amino acid residues, as determined by a given method. Techniques for determining whether an antibody binds to the “same C5 epitope” as the antibody described herein include, for example, epitope mapping methods such as X-ray analysis of the antigen-antibody complex crystals, where atomic analysis of the epitope is provided, and hydrogen / deuterium exchange mass spectrometry (HDX-MS). Other methods may involve monitoring antibody binding to peptide antigen fragments or mutant variations of antigens, where loss of binding due to modification of amino acid residues in the antigen sequence is often considered an expression of the epitope component. Furthermore, computational combinatorial methods for epitope mapping may also be used. These methods rely on the ability of the antibody of interest to affinity isolate specific short peptides from combinatorial phage display peptide libraries. Antibodies having the same VH and VL sequences or the same CDR1, 2, and 3 sequences are expected to bind to the same epitopes.

[0117] An antibody that "competes with another antibody for binding to a target" refers to an antibody that inhibits (partially or completely) the binding of the other antibody to that target. Whether two antibodies compete with each other for binding to a target, i.e., whether one antibody inhibits the binding of the other antibody to the target, and to what extent, can be determined using known competition experiments. In certain embodiments, an antibody competes with another antibody by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% to inhibit the binding of the other antibody to its target. The level of inhibition or competition may vary depending on which antibody is the "inhibiting antibody" (i.e., the cold antibody initially incubated with the target). Competing antibodies bind to the same epitope, overlapping epitopes, or adjacent epitopes (e.g., as is evident from steric hindrance).

[0118] The anti-C5 antibodies, or their antigen-binding fragments, described herein and used in the methods described herein can be produced using a variety of techniques recognized in the art. Monoclonal antibodies can be obtained by a variety of techniques well known to those skilled in the art. Briefly, spleen cells derived from animals immunized with a desired antigen are usually immortalized by fusion with myeloma cells (see Kohler & Milstein, Eur.J.Immunol.6:511-519 (1976)). Alternative immortalization methods include transformation using Epstein-Barr virus, oncogenes, or retroviruses, or other methods well known in the art. Colonies resulting from a single immortalized cell are screened for the production of antibodies with desired specificity and affinity for the antigen, and the production of monoclonal antibodies by such cells may be enhanced by a variety of techniques, including injection into the peritoneal cavity of a vertebrate host. Alternatively, DNA sequences encoding monoclonal antibodies or their binding fragments may be isolated by screening a DNA library from human B cells according to a general protocol outlined by Huse, et al., Science 246:1275-1281 (1989).

[0119] III. Highly concentrated anti-C5 antibody solution This specification provides stable aqueous solutions containing an anti-C5 antibody (e.g., ravulizumab). The aqueous solutions described herein may be sterile, pharmaceutical-grade compositions, for example, for administration to subjects for the treatment or prevention of complement-related disorders such as PNH or aHUS. The solutions described herein can be formulated according to standard methods. Pharmaceutical formulation is a well-established technique, for example, Gennaro (2000) “Remington: The Science and Practice of Pharmacy,” 20 th Edition, Lippincott, Williams & Wilkins (ISBN: 0683306472); Ansel et al. (1999) “Pharmaceutical Dosage Forms and Drug Delivery Systems,” 7 th Edition, Lippincott Williams & Wilkins Publishers (ISBN:0683305727); and Kibbe (2000) “Handbook of Pharmaceutical Excipients American Pharmaceutical Association,”3 rd This is described in detail in Edition (ISBN: 091733096X). Suitable formulation methods for the high-concentration antibody solutions described herein are illustrated in the examples.

[0120] The aqueous solutions described herein contain high concentrations of antibodies that bind to human complement component C5, such as ravulizumab. Such solutions may be referred to herein as “high-concentration antibody solutions.” As used herein, “high concentration” of anti-C5 antibody (e.g., ravulizumab) in an aqueous solution means at least 40 mg / mL or more (for example, at least 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, The antibody concentration is 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, or 300 mg / mL or higher. In one embodiment, the anti-C5 antibody is present in the solution at a concentration greater than 100 mg / mL (e.g., greater than 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, or 195 mg / mL). In another embodiment, the anti-C5 antibody is present in the solution at a concentration greater than 200 mg / mL (e.g., greater than 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, or 295 mg / mL). In yet another embodiment, the anti-C5 antibody is present in the solution at a concentration greater than 300 mg / mL.In another embodiment, the antibody is, for example, 40 mg / mL to 200 mg / mL, 50 mg / mL to 200 mg / mL, 60 mg / mL to 200 mg / mL, 70 mg / mL to 200 mg / mL, 80 mg / mL to 200 mg / mL, 90 mg / mL to 200 mg / mL, 100 mg / mL to 200 mg / mL, 110 mg / mL to 200 mg / mL, 120 mg / mL to 200 mg / mL, 130 mg / mL to 200 mg / mL, 140 mg / mL to 200 mg / mL, 150 mg / mL mL~200mg / mL, 40mg / mL~100mg / mL, 50mg / mL~100mg / mL, 60mg / mL~100mg / mL, 70mg / mL~100mg / mL, 80mg / mL~100mg / mL, 90mg / mL~10 0mg / mL, 40mg / mL~150mg / mL, 50mg / mL~150mg / mL, 60mg / mL~150mg / mL, 70mg / mL~150mg / mL, 80mg / mL~150mg / mL, 90mg / mL~150mg / m L, 100mg / mL~150mg / mL, 110mg / mL~150mg / mL, 120mg / mL~150mg / mL, 40mg / mL~50mg / mL, 40mg / mL~250mg / mL, 50mg / mL~250mg / mL, 6 0mg / mL~250mg / mL, 70mg / mL~250mg / mL, 80mg / mL~250mg / mL, 90mg / mL~250mg / mL, 100mg / mL~250mg / mL, 110mg / mL~250mg / mL, 120m It is present in solution at concentrations of g / mL to 250 mg / mL, 130 mg / mL to 250 mg / mL, 140 mg / mL to 250 mg / mL, 150 mg / mL to 250 mg / mL, 160 mg / mL to 250 mg / mL, 170 mg / mL to 250 mg / mL, 180 mg / mL to 250 mg / mL, 190 mg / mL to 250 mg / mL, 200 mg / mL to 250 mg / mL, greater than 200 mg / mL (e.g., at least 201 mg / mL) to 250 mg / mL, or greater than 200 mg / mL (e.g., 201 mg / mL or more) to 300 mg / mL.

[0121] As described herein and illustrated in the examples, the aqueous solutions of the present invention provide anti-C5 antibodies formulated in the aqueous solution with remarkable physical and chemical stability as well as functional stability. For example, the formulations described herein can maintain the structural integrity of anti-C5 antibodies (e.g., ravulizumab) present in high concentrations in the solution. In one embodiment, the solution is suitable for storage at 2–8°C (e.g., 4°C). In another embodiment, the solution is formulated for storage at temperatures below 0°C (e.g., -20°C or -80°C). In another embodiment, the solution is formulated for storage at 2–8°C (e.g., 4°C) for up to 3 years (e.g., 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 1.5 years, 2 years, 2.5 years, or 3 years). In another embodiment, the solution is suitable for storage at 2–8°C (e.g., 4°C) for at least 1 year, 2 years, or 3 years.

[0122] As illustrated in the examples described herein, the solutions described herein are suitable for maintaining anti-C5 antibodies at a concentration of approximately 100 mg / mL, mainly in monomeric form, for up to two years at approximately 2°C to 8°C. When used herein, anti-C5 antibodies formulated at high concentrations in aqueous solutions of the present invention are such that the antibodies present in the solution are measured, for example, by size exclusion chromatography (SEC-HPLC, e.g., gel permeation HPLC), at least 95% (e.g., at least 95.1, 95.2, 95.3, 95.4, 95.5, 95.6, 95.7, 95.8, 95.9, 96, 96.1, 96.2, 96.3, 96.4, 96) If 0.5, 96.6, 96.7, 96.8, 96.9, 97, 97.1, 97.2, 97.3, 97.4, 97.5, 97.6, 97.7, 97.8, 97.9, 98, 98.1, 98.2, 98.3, 98.4, 98.5, 98.6, 98.7, 98.8, 98.9, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8 or 99.9% or more are monomers, then it is either "primarily monomeric" or "primarily monomeric." In one embodiment, the anti-C5 antibody in the solution described herein can be maintained primarily as monomers after being stored at approximately 2°C to 8°C (e.g., at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 months or more) for at least one month (e.g., at 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10°C).

[0123] In one embodiment of any of the solutions described herein, the anti-C5 antibody (e.g., ravulizumab) retains at least 95% (e.g., at least 96, 97, 98, or 99%) monomers during a storage period of at least 6 months at 2°C to 8°C, as determined by SEC-HPLC (e.g., gel permeation HPLC). In another embodiment, the anti-C5 antibody retains at least 95% (e.g., at least 96, 97, 98, or 99%) monomers during a storage period of at least 9 months at 2°C to 8°C, as determined by SEC-HPLC. In yet another embodiment, the anti-C5 antibody retains at least 95% (e.g., at least 96, 97, 98, or 99%) monomers during a storage period of at least 1 year at 2°C to 8°C, as determined by SEC-HPLC. In yet another embodiment, the anti-C5 antibody retains at least 95% (e.g., at least 96, 97, 98, or 99%) monomers during a storage period of at least 18 months at 2°C to 8°C, as determined by SEC-HPLC. In another embodiment, the anti-C5 antibody retains at least 95% (e.g., at least 96, 97, 98, or 99%) of monomers during a storage period of at least two years at 2°C to 8°C, as determined by SEC-HPLC.

[0124] In another embodiment, less than 5% of the antibody in solution (e.g., 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3, 4.2, 4.1, 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3, 3.2, 3.1, 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1%) is oligomeric, aggregated, and / or fragmented. As used herein, antibody fragmentation refers to improperly assembled components or degradation products of a whole antibody having a molecular weight lower than that of the whole antibody. Such fragmentation forms include, but are not limited to, free monomeric heavy chain polypeptides, dimeric heavy chain polypeptides (e.g., disulfide-bonded heavy chain polypeptides), dimeric heavy chain polypeptides bonded to one light chain polypeptide, monomeric heavy chain polypeptides bonded to one light chain polypeptide, or further degradation products or fragments of a light chain polypeptide or heavy chain polypeptide. In some embodiments, less than 2% of the antibody (e.g., 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1%) agglutinates after being stored at 2°C to 8°C for at least one month (e.g., at least two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nine, twenty, twenty, twelve, twenty-two, twenty-three, or twenty-four months or more). In some embodiments, less than 1% (0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1%) of the antibody fragments after being stored at 2°C to 8°C for at least one month (e.g., at least two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twelve, twelve, twelve, thirty-four months or more).Methods for determining the amount of monomeric antibodies, as well as the amounts of oligomeric, agglutinated, or fragmented anti-C5 antibodies present in a solution, are described herein and illustrated in the examples. For example, those skilled in the art can determine the proportions of total antibodies, fragmented antibodies, unfolding intermediate antibodies, and / or agglutinated antibodies present in a given solution using methods such as size exclusion chromatography (SEC-HPLC, e.g., gel permeation HPLC), static light scattering (SLS), Fourier transform infrared spectroscopy (FTIR), circular dichroism (CD), urea-induced protein unfolding, endogenous tryptophan fluorescence, non-reduced sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and differential scanning calorimetry (DSC).

[0125] In any one embodiment of the solutions described herein, less than 5% of the anti-C5 antibody (e.g., ravulizumab) in the solution is aggregated when determined by SEC-HPLC (e.g., gel permeation HPLC). In another embodiment, less than 4% of the anti-C5 antibody in the solution is aggregated when determined by SEC-HPLC. In yet another embodiment, less than 3% of the anti-C5 antibody in the solution is aggregated when determined by SEC-HPLC. In yet another embodiment, less than 2% of the anti-C5 antibody in the solution is aggregated when determined by SEC-HPLC. In yet another embodiment, less than 1% of the anti-C5 antibody in the solution is aggregated when determined by SEC-HPLC.

[0126] As described herein and illustrated in the examples, the solutions of the present invention containing anti-C5 can retain at least 90% (e.g., 91, 92, 93, 94, 95, 96, 97, 98, 99 or even 100%) of their biological / functional activity (e.g., ability to bind to human C5) after being stored at 2°C to 8°C for at least one month (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36 months or more).

[0127] In another embodiment, an anti-C5 antibody (e.g., ravulizumab) present in the solution described herein can retain at least 90% (e.g., 91, 92, 93, 94, 95, 96, 97, 98, 99 or even 100%) of its hemolytic inhibitory activity after being stored at 2°C to 8°C for at least one month (e.g., at least two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty Suitable hemolysis assays for determining whether the antibody in the solution of the present invention retains its activity are described herein and known in the art, such as in vitro hemolysis assays using avian or porcine red blood cells. Suitable methods for evaluating the ability of antibody preparations to bind to human complement component C5 are known in the art and are described herein.

[0128] In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains its C5-binding activity by at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) for at least 6 months at 2°C to 8°C compared to a reference anti-C5 antibody corresponding to the anti-C5 antibody before storage. In another embodiment, an anti-C5 antibody (e.g., ravulizumab) maintains its C5-binding activity by at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) for at least 9 months at 2°C to 8°C compared to a reference anti-C5 antibody corresponding to the anti-C5 antibody before storage. In another embodiment, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99)% of its C5-binding activity for at least 18 months at 2°C to 8°C, compared to a reference anti-C5 antibody (e.g., ravulizumab) that corresponds to an anti-C5 antibody before storage. In another embodiment, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) of its C5-binding activity for at least 3 years at 2°C to 8°C, compared to a reference anti-C5 antibody (e.g., ravulizumab) that corresponds to an anti-C5 antibody before storage.

[0129] In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99)% of its hemolytic inhibitory ability for at least 6 months at 2°C to 8°C, compared to a reference anti-C5 antibody (e.g., ravulizumab) that corresponds to an anti-C5 antibody before storage. In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99)% of its hemolytic inhibitory capacity for at least 18 months at 2°C to 8°C, compared to a reference anti-C5 antibody (e.g., ravulizumab) that corresponds to an anti-C5 antibody before storage. In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99)% of its hemolytic inhibitory capacity for at least 18 months at 2°C to 8°C, compared to a reference anti-C5 antibody that corresponds to an anti-C5 antibody before storage. In any other embodiment of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) of its hemolytic inhibitory ability for at least two years at 2°C to 8°C, compared to a reference anti-C5 antibody equivalent to the anti-C5 antibody before storage.In another embodiment of any of the solutions described herein, an anti-C5 antibody (e.g., ravulizumab) maintains at least 80% (e.g., at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) of its hemolytic inhibitory ability for at least 3 years at 2°C to 8°C, compared to a reference anti-C5 antibody equivalent to the anti-C5 antibody before storage.

[0130] The aqueous solutions described herein may contain one or more universal agents (e.g., one or more excipients and / or additives such as buffers, sugars or monosaccharides, salts, surfactants, solubilizers, diluents, binders, stabilizers, salts, lipophilic solvents, amino acids, chelators and / or preservatives).

[0131] In one embodiment, the aqueous solution comprises one or more buffers. As used herein, the term “buffer” means one or more components that, when added to an aqueous solution, can protect the solution from pH changes due to the addition of an acid or alkali, or from pH changes during dilution with a solvent. In one embodiment, the solution comprises at least one or more buffers. Non-limiting examples of typical buffers that may be included in a washing solution include Tris (tris(hydroxymethyl)methylamine), bis-Tris, bis-Trispropane, histidine, triethanolamine, diethanolamine, formate, acetate, MES (2-(N-morpholino)ethanesulfonic acid), phosphate, HEPES (4-2-hydroxyethyl-1-piperazineethanesulfonic acid), citrate, MOPS (3-(N-morpholino)propanesulfonic acid), TAPS ( Examples include 3{[tris(hydroxymethyl)methyl]amino}propanesulfonic acid), Bicine (N,N-bis(2-hydroxyethyl)glycine), Tricine (N-tris(hydroxymethyl)methylglycine), TES (2-{[tris(hydroxymethyl)methyl]amino}ethanesulfonic acid), PIPES (piperazine-N,N'-bis(2-ethanesulfonic acid), cacodylate (dimethylarsinic acid), SSC (sodium citrate salts), and sodium phosphate.

[0132] In another embodiment, the buffer is an amino acid. The amino acid may be an amino acid selected from the group consisting of, for example, histidine (e.g., L-histidine), serine (e.g., L-serine), and glycine (e.g., L-glycine). In another embodiment, the solution contains two or more buffers. In a particular embodiment, the buffer is sodium phosphate. In one embodiment, the solution of the present invention does not contain any free amino acids as buffers. In another embodiment, the solution of the present invention contains only one free amino acid (e.g., histidine) as a buffer. In another embodiment, the solution of the present invention may contain two or more (e.g., 2, 3, 4, 5, 6, or 7 or more) different amino acids as buffers, such as serine and histidine.

[0133] The concentration of the buffer is sufficient to maintain the desired pH and may be varied, for example, to maintain the isotonicity of the formulation. Typical concentrations of standard buffers used in parenteral formulations can be found below: Pharmaceutical Dosage Form: Parenteral Medications, Volume 1, 2nd. Edition,Chapter 5,p.194,De Luca and Boylan,“Formulation of Small Volume Parenterals”,Table 5:Commonly used additives in Parenteral Product. In one embodiment, the concentration of one or more buffers in the formulation is about 10 mM to 300 mM, including both ends. In another embodiment, the solution includes at least one buffer at a concentration of 10 mM to 200 mM, including both ends. In another embodiment, the aqueous solution described herein includes a buffer at a concentration of at least 10 mM (e.g., at least 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290 or 300 mM or more). In another embodiment, the aqueous solution is approximately 10mM-50mM, 15mM-50mM, 20mM-50mM, 25mM-50mM, 30mM-50mM, 40mM-50mM, 10mM-100mM, 15mM-100mM, 20mM-100mM, 25mM-100mM, 30mM-100mM, 40mM-100mM, 10mM-150mM, 15mM-1 50mM, 20mM~150mM, 25mM~150mM, 30mM~150mM, 40mM~150mM, 50mM~100mM, 60mM~100mM, 70mM~100 mM, 80mM~100mM, 50mM~150mM, 60mM~150mM, 70mM~150mM, 80mM~150mM, 90mM~150mM, 100mM~150m M, 10mM~200mM, 15mM~200mM, 20mM~200mM, 25mM~200mM, 30mM~200mM, 40mM~200mM, 50mM~200mM, 60mM~200mM, 70mM~200mM, 80mM~200mM, 90mM~200mM, 100mM~200mM, 150mM~200mM, 10mM~250mM, Contains buffer at concentrations of 15mM-250mM, 20mM-250mM, 25mM-250mM, 30mM-250mM, 40mM-250mM, 50mM-250mM, 60mM-250mM, 70mM-250mM, 80mM-250mM, 90mM-250mM, 100mM-250mM, 150mM-250mM, or 200mM-250mM.In another embodiment, the concentration of the buffer in the formulation is about 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 90 mM, 95 mM, or about 100 mM. In another embodiment, the buffer is present in the solution at a concentration of 20 mM or higher. In another embodiment, the buffer is present in the solution at a concentration of 25 mM or higher. In another embodiment, the buffer is present in the solution at a concentration of 50 mM or higher. In embodiments of the present invention, the solution contains two or more different buffers (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more), each of the two or more buffers may be present independently at any one of the concentrations described above.

[0134] In one embodiment, the aqueous solution may have a neutral pH or be prepared to have a neutral pH. As used herein, “neutral pH” is a pH of 7 to 8, including both ends. Thus, as used herein, neutral pH includes specific pH values ​​such as 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, and 8.0. In some embodiments, neutral pH is at least pH 7 (e.g., at least pH 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.7, or 7.9) but less than pH 8 (e.g., pH 7.9, 7.8, 7.7, 7.6, 7.5, 7.4, 7.3, 7.2, or less than 7.1). Thus, in some embodiments, neutral pH may be at least pH 7 but less than pH 7.5. In some embodiments, neutral pH may be between pH 7 and pH 7.5. In some embodiments, the neutral pH may be pH 7 to pH 7.2. In other embodiments, the pH of the solution is 7.0 to 7.4. In other embodiments, the pH of the solution is 7.2 to 7.8. In other embodiments, the pH of the solution is 7.2 to 7.6. In some embodiments, the neutral pH may be, for example, pH 7. Those skilled in the art will further recognize that human blood (e.g., human blood from a healthy subject) has a neutral pH as defined herein, for example, the pH of human blood is approximately pH 7.35 to pH 7.45. See, for example, Boron and Boulpaep (2003) “Medical physiology: a cellular and molecular approach,” WBSaunders, New York (ISBN: 0721632564). In some embodiments, the pH of the highly concentrated antibody solution described herein is approximately 6.4 to 7.5 (e.g., approximately 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, or 7.7), including both ends. In one embodiment, the pH of the solution is 7.2 to 7.6. In a particular embodiment, the pH of the solution is 7.4.

[0135] In one embodiment, the solution comprises one or more surfactants, such as anionic, cationic, or nonionic surfactants. As used herein, the term “surfactant” refers to a surface-active molecule that contains both a hydrophobic moiety (e.g., an alkyl chain) and a hydrophilic moiety (e.g., a carboxyl group and a carboxylic acid group). The surfactants suitable for use in the formulation of the present invention are not limited to, but include fatty acid esters (e.g., sorbitan monocaprate, sorbitan monolaurate, sorbitan monopalmitate), sorbitan trioleates, glycerin fatty acid esters (e.g., glycerin monocaprate, glycerin monomyristate, glycerin monostearate), polyglycerin fatty acid esters (e.g., decaglyceryl monostearate, decaglyceryl distearate, decaglyceryl monolinoleate), polyoxyethylene sorbitan fatty acid esters (e.g., sorbitan polyoxyethylene monolaurate, sorbitan polyoxyethylene monooleate, sorbitan polyoxyethylene monostearate, sorbitan polyoxyethylene monopalmitate, sorbitan polyoxyethylene trioleate, sorbitan polyoxyethylene tristearate), and polyoxyethylene sorbitol fatty acid esters (e.g., polyoxyethylene sorbitol Sorbitol lastearate, sorbitol polyoxyethylene tetraoleate), polyoxyethylene glycerin fatty acid ester (e.g., glyceryl polyethylene monostearate), polyethylene glycol fatty acid ester (e.g., glycol polyethylene distearate), polyoxyethylene alkyl ester (e.g., polyoxyethylene lauryl ether), polyoxyethylene polyoxypropylene alkyl ether (e.g., polyoxyethylene polyoxypropylene glycol, polyoxyethylene polyoxypropylene propyl ether, polyoxyethylene polyoxypropylene cetyl ether), polyoxyethylene alkylphenyl ether (e.g., polyoxyethylene nonylphenyl ether), polyoxyethylene hydrogenated castor oil (e.g., polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil), polyoxyethylene beeswax derivative (e.g., polyoxyethylene sorbitol beeswax),Examples include polyoxyethylene lanolin derivatives (e.g., polyoxyethylene lanolin) and polyoxyethylene fatty acid amides (e.g., polyoxyethylene stearate amide); C12-C18 alkyl sulfates (e.g., sodium cetyl sulfate, sodium lauryl sulfate, sodium oleyl sulfate); polyoxyethylene C10-C18 alkyl ether sulfates with an average of 2-4 moles of ethylene oxide units added (e.g., sodium polyoxyethylene lauryl sulfate and C10-C18 alkyl sulfosuccinate esters (e.g., sodium lauryl sulfosuccinate ester)); and natural surfactants such as lecithin, glycerophospholipids, and sphingophospholipids (e.g., sphingomyelin); and sucrose esters of C12-C18 fatty acids.

[0136] In one embodiment, the surfactant in the formulation is a nonionic surfactant. In a particular embodiment, the surfactant in the formulation is a polyoxyethylene sorbitan fatty acid ester, such as polysorbate 20, 40, 60, 80, or one or more combinations thereof. In one embodiment, the surfactant in the formulation is polysorbate 80 (Tween 80). In another embodiment, the surfactant in the formulation is polysorbate 60. In another embodiment, the surfactant in the formulation is polysorbate 40. In another embodiment, the surfactant in the formulation is polysorbate 20 (Tween 20).

[0137] The amount of surfactant added to the formulation is sufficient to reduce aggregation of the formulated antibody and / or to minimize the formation of particulate matter in the formulation. For example, the surfactant may be present in the formulation in an amount of about 0.001% to about 1%, or about 0.001% to about 0.5%, or about 0.01% to about 0.2%. In one embodiment, the aqueous solution is at least, or approximately, 0.001 (e.g., at least, or approximately 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, Contains surfactants at concentrations of 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.3, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, or 0.5% or higher. In another embodiment, the aqueous solution contains 0.2% (e.g., 0.19, 0.18, 0.17, 0.16, 0.15, 0.14, 0.13, 0.12, 0.11, 0.10, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.002, or 0.001)% or less of a pharmaceutically acceptable surfactant.

[0138] In another embodiment, the formulation contains polysorbate at a concentration of about 0.001% to about 0.5%, about 0.005% to about 0.2%, about 0.01% to about 0.1%, or about 0.02% to about 0.06%, or about 0.03% to about 0.05% (w / v). In a particular embodiment, the formulation contains polysorbate at a concentration of 0.01%, or 0.02%, or 0.03%, or 0.04%, or 0.05%, or 0.06%, or 0.07%, or 0.08%, or 0.09%, or 0.1%, or 0.15%, or 0.2% (w / v). In a particular embodiment, the surfactant is present in the formulation at an amount of 0.02% or about 0.04% (w / v). In one embodiment, the surfactant is present in the formulation at an amount of 0.05% (w / v).

[0139] In one embodiment, the formulation comprises at least about 0.01%, at least about 0.02%, at least about 0.05%, at least about 0.1%, at least about 0.2%, at least about 0.3%, at least about 0.4%, or at least about 0.5% of polysorbate 80. In certain embodiments, the formulation contains about 0.01% to about 0.5%, about 0.01% to about 0.3%, about 0.001% to about 0.2%, about 0.02% to about 0.5%, about 0.02% to about 0.3%, about 0.02% to about 0.2%, about 0.05% to about 0.5%, about 0.05% to about 0.3%, about 0.05% to about 0.2%, about 0.075% to about 0.5%, about 0.075% to about 0.3%, or about 0.075% to about 0.2% of polysorbate 80. In further embodiments, the formulation contains about 0.01%, about 0.02%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, or about 0.5% of polysorbate 80. In one embodiment, the formulation contains about 0.05% polysorbate 80. In one embodiment, the formulation contains about 0.04% polysorbate 80. In one embodiment, the formulation contains about 0.03% polysorbate 80. In one embodiment, the formulation contains about 0.02% polysorbate 80. In one embodiment, the formulation contains about 0.01% polysorbate 80.

[0140] In one embodiment, the aqueous solution contains one or more salts, such as sodium chloride, potassium chloride, or magnesium chloride. In some embodiments, the aqueous solutions described herein contain salts at a concentration of at least 10 mM (for example, at least 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, or 300 mM or more). In some embodiments, the aqueous solutions described herein may contain salts at concentrations of approximately 200 mM or less (for example, approximately 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 90, 80, 70, 60, 50, 40, 30, 25, 20, 15 or 10 mM or less).In some embodiments, the aqueous solutions described herein are approximately 10mM-50mM, 15mM-50mM, 20mM-50mM, 25mM-50mM, 30mM-50mM, 40mM-50mM, 10mM-100mM, 15mM-100mM, 20mM-100mM, 25mM-100mM, 30mM-100mM, 40mM-100mM, 10mM-150mM M, 15mM~150mM, 20mM~150mM, 25mM~150mM, 30mM~150mM, 40mM~150mM, 50mM~100mM, 60mM~100mM, 7 0mM~100mM, 80mM~100mM, 50mM~150mM, 60mM~150mM, 70mM~150mM, 80mM~150mM, 90mM~150mM, 100m M~150mM, 10mM~200mM, 15mM~200mM, 20mM~200mM, 25mM~200mM, 30mM~200mM, 40mM~200mM, 50mM~2 00mM, 60mM~200mM, 70mM~200mM, 80mM~200mM, 90mM~200mM, 100mM~200mM, 150mM~200mM, 10mM~25 The solution contains salts at concentrations of 0 mM, 15 mM to 250 mM, 20 mM to 250 mM, 25 mM to 250 mM, 30 mM to 250 mM, 40 mM to 250 mM, 50 mM to 250 mM, 60 mM to 250 mM, 70 mM to 250 mM, 80 mM to 250 mM, 90 mM to 250 mM, 100 mM to 250 mM, 150 mM to 250 mM, or 200 mM to 250 mM. In embodiments of the present invention, the solution contains two or more different salts (for example, at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more), each of the two or more salts may exist independently at any one of the concentrations described above.

[0141] In one embodiment, the aqueous solution comprises one or more carbohydrate excipients. Suitable carbohydrate excipients are, for example, Katakam and Banga (1995) J Pharm Pharmacol. 47(2) :103-107;Andya et al.(2003)AAPS PharmSci 5(2)See Article 10; and Shire (2009) “Current Trends in Monoclonal Antibody Development and Manufacturing,” Volume 11, Springer, page 354. Suitable carbohydrate excipients for use in the solutions described herein include, but are not limited to, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, and sorbose; disaccharides such as lactose, sucrose, trehalose, and cellobiose; polysaccharides such as maltodextrin, dextran, and starch; and sugar alcohols such as mannitol, xylitol, maltitol, lactitol, and sorbitol. In one embodiment, the carbohydrate excipient is present in the solution of the present invention at a concentration of at least or approximately 0.5% (e.g., at least or approximately 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10% or more). In embodiments in which the solution of the present invention comprises two or more different carbohydrate excipients (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more), each excipient may be present independently at any of the concentrations described above.

[0142] In another embodiment, the stable aqueous solution contains one or more stabilizers. Examples of stabilizers, but not limited to, include polyols, sugars (e.g., sucrose or trehalose), amino acids (e.g., arginine), amines, and salting-out salts. In one embodiment, the solution contains both ends and at least one stabilizer at a concentration of 2–10%. In one embodiment, the solution contains 5% sucrose. In another embodiment, the solution contains both ends and at least one or more stabilizers at a concentration of 10 mM–50 mM. In another embodiment, the stabilizer is present in the solution at a concentration of 20 mM or higher. In another embodiment, the stabilizer is present in the solution at a concentration of at least 25 mM or 25 mM. In another embodiment, the stabilizer is present in the solution at a concentration of 50 mM or higher. In another embodiment, the solution contains 25 mM arginine.

[0143] In one embodiment, the solution described herein comprises one or more preservatives.

[0144] As used herein, the term “preservative” refers to an agent that reduces the action of bacteria and may optionally be added to the formulations herein. The addition of preservatives may facilitate the manufacture of, for example, multi-use (multiple-dose) formulations. Examples of possible preservatives include octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride (a mixture of alkylbenzyldimethylammonium chloride, where the alkyl group is a long-chain compound), and benzethonium chloride. Other types of preservatives include aromatic alcohols such as phenol, butyl alcohol, and benzyl alcohol, alkylparabens such as methylparaben or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol.

[0145] In one embodiment, the stable aqueous solution contains five or fewer agents in addition to the anti-C5 antibody. In another embodiment, the stable aqueous solution contains four or fewer agents in addition to the anti-C5 antibody. In yet another embodiment, the stable aqueous solution contains three or fewer agents in addition to the anti-C5 antibody. In yet another embodiment, the stable aqueous solution contains two or fewer agents in addition to the anti-C5 antibody. In yet another embodiment, the stable aqueous solution contains one or fewer agents in addition to the anti-C5 antibody.

[0146] In one embodiment, the stable aqueous solution contains a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and an amino acid sequence described in SEQ ID NO: 6. Anti-C5 antibody containing a light chain CDR3 with a minoic acid sequence, 50±15 mM (e.g., 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or 65) mM phosphate buffer, 5±3 (e.g., 2, 3, 4, 5, 6, 7 or 8) % sucrose , and also comprising 25±10 (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 or 35) mM of arginine, in which case the solution has a pH of 7.4±0.5 (e.g., 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8 or 7.9).

[0147] In another embodiment, the stable aqueous solution contains a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and an amino acid sequence described in SEQ ID NO: 6, all at a concentration of 100 ± 20 mg / mL (e.g., 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120) mg / mL, a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and an amino acid sequence described in SEQ ID NO: 6 Anti-C5 antibody containing a light chain CDR3 containing an acid sequence, 50±15 mM (e.g., 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or 65) mM phosphate buffer, 5±3 (e.g., 2, 3, 4, 5, 6, 7 or 8) % sucrose, The solution consists of 25 ± 10 mM (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) mM arginine, in which case the solution has a pH of 7.4 ± 0.5 (e.g., 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, or 7.9).

[0148] In another embodiment, the stable aqueous solution has a concentration of 100 ± 20 mg / mL (e.g., 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120) Anti-C5 antibody containing heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6, 50±15 (for example) 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or 65) mM phosphate buffer, 5 ± 3 (e.g., 2, 3, 4, 5, 6, 7 or 8) % sucrose, 25 ± 10 (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 2 The solution comprises 7, 28, 29, 30, 31, 32, 33, 34, or 35 mM arginine, and 0.05 ± 0.03 (e.g., 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, and 0.08)% polysorbate 80, in which case the solution has a pH of 7.4 ± 0.5 (e.g., 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, or 7.9).

[0149] In another embodiment, the stable aqueous solution is a mixture of 100 ± 20 mg / mL (e.g., 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120) mg / mL. Anti-C5 antibody containing heavy chain CDR1 containing the amino acid sequence described in sequence number 19, heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6, 50±15 (for example) 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or 65) mM phosphate buffer, 5 ± 3 (e.g., 2, 3, 4, 5, 6, 7 or 8) % sucrose, 25 ± 10 (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 The solution comprises 28, 29, 30, 31, 32, 33, 34 or 35 mM arginine and 0.05 ± 0.03 (e.g., 0.02, 0.03, 0.04, 0.05, 0.06, 0.07 and 0.08)% polysorbate 80, in which case the solution has a pH of 7.4 ± 0.5 (e.g., 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8 or 7.9).

[0150] In another embodiment, a stable aqueous solution is provided (e.g., a sterile solution) in which the solution comprises (a) an anti-C5 antibody (e.g., ravulizumab) at a concentration of about 100 mg / mL, (b) about 50 mM phosphate buffer, (c) about 5% sucrose, and (d) about 25 mM arginine. In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody (e.g., ravulizumab) at a concentration of 100 mg / mL, (b) 50 mM phosphate buffer, (c) 5% sucrose, and (d) 25 mM arginine.

[0151] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of about 100 mg / mL, (b) a phosphate buffer at about 50 mM, (c) sucrose at about 5%, (d) polysorbate 80 at about 0.05%, and (e) arginine at about 25 mM.

[0152] In another embodiment, the stable aqueous solution contains three or fewer additional agents. In another embodiment, the stable aqueous solution contains two or fewer additional agents. In another embodiment, the stable aqueous solution contains one or fewer additional agents.

[0153] In another embodiment, the stable aqueous solution consists of (a) an anti-C5 antibody at a concentration of about 100 mg / mL, (b) a phosphate buffer at about 50 mM, (c) sucrose at about 5%, and (d) arginine at about 25 mM.

[0154] In another embodiment, the stable aqueous solution consists of (a) an anti-C5 antibody at a concentration of about 100 mg / mL, (b) about 50 mM phosphate buffer, (c) about 5% sucrose, (d) about 0.05% polysorbate 80, and (e) about 25 mM arginine.

[0155] In one embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of about 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) about 50 mM phosphate buffer; (c) about 5% sucrose; and (d) about 25 mM arginine.

[0156] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) 50 mM phosphate buffer; (c) 5% sucrose; and (d) 25 mM arginine.

[0157] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of about 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) about 50 mM phosphate buffer; (c) about 5% sucrose; (d) about 0.05% polysorbate 80; and (e) about 25 mM arginine.

[0158] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) 50 mM phosphate buffer; (c) 5% sucrose; (d) 0.05% polysorbate 80; and (e) about 25 mM arginine.

[0159] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of about 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) about 50 mM phosphate buffer; (c) about 5% sucrose; (d) about 0.05% polysorbate 80; and (e) about 25 mM arginine.

[0160] In another embodiment, the stable aqueous solution comprises (a) an anti-C5 antibody at a concentration of 100 mg / mL, wherein the anti-C5 antibody comprises a heavy chain CDR1 containing the amino acid sequence described in SEQ ID NO: 19, a heavy chain CDR2 containing the amino acid sequence described in SEQ ID NO: 18, a heavy chain CDR3 containing the amino acid sequence described in SEQ ID NO: 3, a light chain CDR1 containing the amino acid sequence described in SEQ ID NO: 4, a light chain CDR2 containing the amino acid sequence described in SEQ ID NO: 5, and a light chain CDR3 containing the amino acid sequence described in SEQ ID NO: 6; (b) 50 mM phosphate buffer; (c) 5% sucrose; (d) 0.05% polysorbate 80; and (e) 25 mM arginine.

[0161] IV. Method for preparing highly concentrated antibody solution This specification also provides a method for preparing a highly concentrated anti-C5 antibody solution. In one embodiment, a method is provided for preparing a stable concentrated antibody solution comprising 100 mg / mL of anti-C5 antibody, 50 mM phosphate buffer, 5% sucrose, and 25 mM arginine, the method being described as follows: i) To provide a first aqueous solution containing an anti-C5 antibody, wherein the first aqueous solution has a first formulation and contains an anti-C5 antibody at a concentration of 10 mg / mL or less. ii) Dialysis filtration is performed on the first aqueous solution to obtain a formulation containing 50 mM phosphate buffer, 5% sucrose, and 25 mM arginine at pH 7.4, thereby producing a second aqueous solution, wherein the second aqueous solution has the second formulation as a result of the dialysis filtration, and iii) The second aqueous solution is concentrated to prepare a stable concentrated antibody solution containing 100 mg / mL of anti-C5 antibody, 50 mM phosphate buffer, 5% sucrose, and 25 mM arginine.

[0162] In another embodiment, a method is provided for preparing a stable concentrated antibody solution comprising 100 mg / mL of anti-C5 antibody, 50 mM phosphate buffer, 5% sucrose, 25 mM arginine, and 0.05% polysorbate 80, the method being described as follows: i) To provide a first aqueous solution containing an anti-C5 antibody, wherein the first aqueous solution has a first formulation and contains an anti-C5 antibody at a concentration of 10 mg / mL or less. ii) Dialysis filtration is performed on the first aqueous solution to a formulation containing 50 mM phosphate buffer, 5% sucrose, 25 mM arginine, and 0.05% polysorbate 80 at pH 7.4, thereby producing a second aqueous solution, wherein the second aqueous solution has the second formulation as a result of the dialysis filtration, and iii) The second aqueous solution is concentrated to prepare a stable concentrated antibody solution containing 100 mg / mL of anti-C5 antibody, 50 mM phosphate buffer, 5% sucrose, 25 mM arginine, and 0.05% polysorbate 80.

[0163] V. Route of administration The solutions described herein can be administered to patients by a variety of methods, the methods of which depend in part on the route of administration. The route may be parenteral, for example, intravenous injection or infusion (IV), subcutaneous injection (SC), intraperitoneal injection (IP), intraocular injection, intra-articular injection, or intramuscular injection (IM). As used herein, “parenteral administration,” “administered parenterally,” and other grammatically equivalent terms refer to modes of administration other than intestinal and local administration, which are usually performed by injection, and include, but are not limited to, intravenous, intranasal, intraocular, transpulmonary, intramuscular, intraarterial, intrathecal, intra-articularsternal, intra-cerebral, intra-cranial, intra-sternal, and intra-sternal injections and infusions.

[0164] In certain embodiments, the solution is administered via subcutaneous injection. Subcutaneous administration can be carried out by a device. The device may be a syringe, a pre-filled syringe, a disposable or reusable auto-injector, a pen injector, a patch injector, a wearable injector, a walk-behind syringe infusion pump with a subcutaneous infusion set, or other devices.

[0165] In one embodiment, the solution described herein is delivered to a target via topical administration. As used herein, “topical administration” or “topical delivery” means delivery that does not rely on the transport of the composition or activator (e.g., anti-C5 antibody) to the target tissue or site of interest via the vascular system. After topical administration around the target tissue or site of interest, the solution, or one or more of its components, may diffuse into the target tissue or site of interest.

[0166] For example, the solution may be delivered by injection or by implantation of a device containing the solution. The implant may be a porous, non-porous, or gelatinous material, including a membrane or fiber such as a silicone rubber membrane. The implant may be configured for continuous or periodic release of the solution to a target. See, for example, U.S. Patent Application Publication 20080241223, U.S. Patents 5,501,856, 4,863,457 and 3,710,795, EP 488401 and EP 430539. Each of these publications is incorporated herein by reference in its entirety. The solutions described herein may be delivered to a target via implantable devices based on, for example, diffusive, erosive, or convective systems, such as osmotic pumps, biodegradable implants, electrodiffusion systems, electroosmotic systems, vapor pressure pumps, electrolytic pumps, foaming pumps, piezoelectric pumps, erosive systems, or electromechanical systems.

[0167] In one embodiment, the solutions described herein can be administered topically to a joint (e.g., an articular joint). For example, in embodiments where the disorder is arthritis, a therapeutically suitable solution can be administered directly to the joint (e.g., intra-articular) or around the joint. Examples of intra-articular joints to which the compositions described herein can be administered topically include, for example, the hip, knee, elbow, wrist, sternoclavicular, temporomandibular joint, carpal tunnel joint, tarsal joint, ankle joint, and any other joints that become arthritis-prone. The compositions described herein can also be administered to sacs such as, for example, the acromion, biceps radial, cubitoradial, deltoid muscle, patellar synovial fluid, ischium, and any other sacs known in the medical field.

[0168] In another embodiment, the solution described herein may be administered topically to the eye. As used herein, the term “eye” refers to any and all anatomical tissues and structures associated with the eye. In one embodiment, the solution described herein is administered into the posterior chamber of the eye. In another embodiment, the solution described herein is administered into the vitreous humor. In another embodiment, the solution described herein is administered transsclerally.

[0169] In some embodiments, for example, embodiments relating to the treatment or prevention of disorders such as COPD or asthma, the solutions described herein may be administered to a subject via the lungs. Pulmonary drug delivery may be carried out by inhalation, and administration by inhalation as herein may be oral and / or nasal. In one embodiment, the solutions described herein may be administered to the lungs of a subject via a nebulizer. The nebulizer uses compressed air to deliver the compound as a liquefied aerosol or mist. The nebulizer may be, for example, a jet nebulizer (e.g., an air or liquid-jet nebulizer) or an ultrasonic nebulizer. Additional devices and intrapulmonary administration methods are described, for example, in U.S. Patent Application Publications 20050271660 and 20090110679. The contents of each of those disclosures are incorporated herein by reference in their entirety.

[0170] In another embodiment, the solutions described herein exist in unit dosage forms, which are particularly suitable for self-administration. The formulation products of this disclosure may be contained in containers, typically such as vials, cartridges, pre-filled syringes, or disposable pens. Administration devices, such as the administration device described in U.S. Patent No. 6,302,855, may also be used. The injection system may include the delivery pen described in U.S. Patent No. 5,308,341. Pen devices are most commonly used for self-delivery of insulin to diabetic patients and are well known in the art. Such devices may include at least one injection needle (e.g., a 31-gauge needle about 5–8 mm in length) and are typically pre-filled with one or more therapeutic unit doses of the solution, which are useful for rapidly delivering the solution to the target with as little pain as possible.

[0171] VI. Treatment method The described solutions can be used to treat a variety of diseases and conditions in human patients. In one embodiment, the solution can be used to treat complement-related disorders, including, but not limited to, rheumatoid arthritis (RA), antiphospholipid syndrome, lupus nephritis, ischemia-reperfusion injury, atypical hemolytic uremic syndrome (aHUS), typical or infectious hemolytic uremic syndrome (tHUS), dense deposit disease (DDD), paroxysmal nocturnal hemoglobinuria (PNH), neuromyelitis optica (NMO), multifocal motor neuropathy (MMN), multiple sclerosis (MS), macular degeneration (e.g., age-related macular degeneration (AMD)), hemolytic, elevated liver enzymes and thrombocytopenia (HELLP) syndrome, thrombotic thrombocytopenic purpura (TTP), spontaneous fetal loss, pauci-immune vasculitis, epidermolysis bullosa, recurrent miscarriage, and traumatic brain injury (e.g., Holers (2008) Immunological Reviews). 223 :300-316 and Holers and Thurman(2004)Molecular Immunology 41 (See pages 147-152).

[0172] In another embodiment, complement-related disorders include, but are not limited to, diabetes-related (e.g., ocular) vascular disorders, central retinal vein occlusion, cardiovascular disorders, myocarditis, cerebrovascular disorders, peripheral (e.g., musculoskeletal) vascular disorders, renal vascular disorders, mesenteric / intestinal vascular disorders, recanalization for transplantation and / or re-transplantation, vasculitis, Henoch-Schönlein purpura nephritis, systemic lupus erythematosus-associated vasculitis, rheumatoid arthritis-associated vasculitis, immune complex vasculitis, Takayasu's arteritis, dilated cardiomyopathy, diabetic vascular disorders, Kawasaki disease (arteritis), venous gas embolism (VGE), and complement-related vascular disorders such as restenosis after stent placement, rotational atherectomy, and percutaneous transcatheter angioplasty (PTCA) (see, for example, U.S. Patent Application Publication 20070172483).

[0173] Disorders associated with additional complement include, but are not limited to, myasthenia gravis, cold agglutinin disease, dermatomyositis, Graves' disease, atherosclerosis, Alzheimer's disease, Guillain-Barré syndrome, Degoss disease, graft rejection (e.g., transplant rejection), sepsis, burns (e.g., severe burns), sepsis with systemic inflammatory response, septic shock, spinal cord injury, glomerulonephritis, Hashimoto's thyroiditis, type 1 diabetes mellitus, psoriasis, pemphigus, autoimmune hemolytic anemia (AIHA), idiopathic thrombocytopenic purpura (ITP), Goodpasture syndrome, antiphospholipid syndrome (APS), fulminant APS (CAPS), amyotrophic lateral sclerosis (ALS), Alzheimer's disease, and chronic inflammatory demyelinating polyneuropathy.

[0174] In another embodiment, the solutions described herein can be used to treat thrombotic microangiopathy (TMA), such as TMA associated with complement-related disorders, including any of the complement-related disorders described herein.

[0175] Complement-related disorders include, but are not limited to, asthma, bronchitis, chronic obstructive pulmonary disease (COPD), interstitial lung disease, alpha-1 antitrypsin deficiency, emphysema, bronchiectasis, bronchiolitis obstructive, alveolitis, sarcoidosis, pulmonary fibrosis, and collagen vascular disease, among other complement-related lung disorders.

[0176] In another embodiment, the solutions described herein are administered to a subject to treat, prevent or improve at least one symptom of a complement-related inflammatory response (e.g., a complement-related inflammatory response pattern of complement-related disorder). For example, the compositions can be used to treat, prevent and / or improve one or more symptoms associated with complement-related inflammatory responses, such as graft rejection / graft-versus-host disease (GVHD), reperfusion injury (e.g., after cardiopulmonary bypass or tissue transplantation), and tissue injury following other types of trauma, such as burns (e.g., severe burns), blunt injuries, spinal cord injuries, or frostbite. For example, Park et al. (1999) Anesth Analg 99(1) :42-48;Tofukuji et al.(1998)J Thorac Cardiovasc Surg 116(6) :1060-1068;Schmid et al.(1997)Shock 8(2) :119-124; and Bless et al. (1999) Am J Physiol 276(1) See lines L57-L63.

[0177] In another embodiment, complement-mediated disorders include, but are not limited to, cardiovascular disorders, myocarditis, cerebrovascular disorders, peripheral (e.g., musculoskeletal) vascular disorders, renal vascular disorders, mesenteric / intestinal vascular disorders, recanalization for transplantation and / or re-transplantation, vasculitis, Henoch-Schönlein purpura nephritis, systemic lupus erythematosus-associated vasculitis, rheumatoid arthritis-associated vasculitis, immune complex vasculitis, organ or tissue transplantation, Takayasu's arteriovenous arthritis, capillary leak syndrome, dilated cardiomyopathy, diabetic vascular disorders, thoracic-abdominal aortic aneurysm, Kawasaki disease (arteritis), venous gas embolism (VGE), and complement-mediated vascular disorders such as restenosis after stent placement, rotational atherectomy, and percutaneous transcatheter coronary angioplasty (PTCA) (see, for example, U.S. Patent Application Publication 20070172483).

[0178] VII. Combination therapy In one embodiment, the solution described herein is administered to a patient as monotherapy. In another embodiment, the solution described herein is administered in combination with one or more additional agents and / or other treatments (e.g., those suitable for the treatment of complement-related disorders). For example, combination therapy may include administering one or more additional agents (e.g., anticoagulants, antihypertensives, or anti-inflammatory agents (e.g., steroids)) to a human patient to provide therapeutic benefits to the patient. In one embodiment, the solution described herein is administered in combination with an anti-inflammatory agent (e.g., NSAIDs, corticosteroids, methotrexate, hydroxychloroquine, anti-TNF agents (e.g., etanercept and infliximab), B-cell depletion agents (e.g., rituximab), interleukin-1 antagonists, or T-cell costimulators (e.g., abatacept)).

[0179] Additional agents for treating complement-related disorders in the patient population vary depending on the specific disorder being treated and are not limited to, but may include one or more antihypertensive agents (e.g., angiotensin-converting enzyme inhibitors, labetalol, hydralazine, nifedipine, calcium channel antagonists, nitroglycerin, or sodium nitroferricyanide), anticoagulants, corticosteroids (e.g., prednisone), immunosuppressants (e.g., vincristine or cyclosporine A), anticoagulants (e.g., warfarin (Coumadin), aspirin, heparin, phenindione, fondaparinux, hydraparinux), thrombin inhibitors (e.g., argatroban, repiridine, bivalirudine, or dabigatran), and fibrinolytic agents (e.g., Anklod, α-aminobutyric acid). Examples include pronates (e.g., antiplasmin-α1, prostacyclin, and defibrotide), antihypertensives (e.g., labetalol, hydralazine, nifedipine, calcium channel antagonists, nitroglycerin, or sodium nitroferricyanide), lipid-lowering agents (e.g., inhibitors of hydroxymethylglutaryl-CoA reductase), anticonvulsants (e.g., magnesium sulfate), antithrombotic agents (e.g., heparin, antithrombin, prostacyclin, or low-dose aspirin), sympathomimetic agents (e.g., albuterol), antibiotics, deoxyribonucleases (e.g., Pulmozyme®), anticholinergics, anti-IgE inhibitors (e.g., anti-IgE antibodies), corticosteroids, or nonsteroidal anti-inflammatory drugs (NSAIDs). Many different NSAIDs are available.Some are available over-the-counter, including ibuprofen (Advil®, Motrin®, Nuprin®) and naproxen (Alleve®), while many others are available by prescription, such as melxicam (Mobic®), etodolac (Lodine®), nabumetone (Relafen®), sulindac (Clinoril®), tolementin (Tolectin®), and salicylate. Examples include magnesium phosphate (Trilasate®), diclofenac (Cataflam®, Voltaren®, Arthrotec®), diflusinal (Dolobid®), indomethacin (Indocin®), ketoprofen (Orudis®, Oruvail®), oxaprozin (Daypro®), and piroxicam (Feldene®) (e.g., Mihu et al. (2007) J Gastrointestin Liver Dis.). 16(4) (See 419-424). In another embodiment, the solutions described herein may be formulated for administration to a patient in conjunction with intravenous gamma globulin therapy (IVIG), plasma exchange, plasma replacement therapy, or plasma exchange therapy.

[0180] In one embodiment, the solution and one or more additional agents and / or treatments are administered simultaneously. In another embodiment, the solution is administered before the administration of one or more additional agents and / or treatments. In yet another embodiment, the solution is administered after the administration of one or more additional agents and / or treatments.

[0181] When the antibody solution described herein is used in combination with a second activator, the agent (e.g., anti-C5 antibody and the second agent) may be formulated separately or together. For example, the solution and the agent may be mixed, for example, immediately before administration, and may be administered together or separately, at the same time or at different times.

[0182] VIII. Kits and individual dosage forms This specification also provides a kit comprising a stable aqueous solution containing an anti-C5 antibody or its antigen-binding fragment, such as ravulizumab or BNJ421, in a therapeutically effective dose suitable for administration to a human patient (e.g., a patient with complement-related disorder). The kit may optionally include instructions, such as an administration schedule, which enable a person (e.g., a physician, nurse, or patient) to administer the composition contained therein, and the solution to be administered to the patient.

[0183] The kit may also include appropriate means for delivering one or more solutions to patients who need them, such as patients suffering from complement-related disorders, patients suspected of having complement-related disorders, or patients at risk of developing complement-related disorders. In one embodiment, the means is suitable for invasive solution delivery to the patient (e.g., intravascular (e.g., intravenous), subcutaneous, intra-articular, intraocular, intravitreous, or intramuscular). In another embodiment, the means is suitable for subcutaneous delivery of the solution to the patient. In yet another embodiment, the means is suitable for intravenous delivery of the solution to the patient. For example, the means may be a syringe or an osmotic pump. In yet another embodiment, the solution may be formulated as eye drops, and the means may be an eye dropper.

[0184] Optionally, the kit may include multiple packages of single-dose solution, each containing an effective amount of solution for a single dose. The kit may also include the apparatus or devices necessary for administering the solution. For example, the kit may provide one or more pre-filled syringes containing the solution.

[0185] The following examples are merely illustrative and should not be construed as in any way limiting the scope of the present disclosure, as numerous variations and equivalents will be apparent to those skilled in the art upon reading the present disclosure.

[0186] The contents of all references, Genbank entries, patents and published patent applications cited throughout this specification are hereby expressly incorporated by reference into this specification.

Example

[0187] Example 1: Development of a High-Concentration Formulation of ALXN1210 for Subcutaneous Administration This example summarizes the development of a high-concentration formulation of ALXN1210 for subcutaneous administration (e.g., at 100 mg / mL, in 50 mM phosphate buffer, 5% sucrose, 25 mM arginine, pH 7.4). Preliminary experiments were conducted in the initial formulation development to obtain pre-formulation screening data for ALXN1210 and to evaluate the decrease at higher concentrations in the milky white appearance. The initial formulation of ALXN1210 (10 mM phosphate, 150 mM sodium chloride, pH 7.0, 0.02% Tween 80, 10 mg / mL) was colorless and slightly milky white. As the concentration of ALXN1210 increased, the milky white appearance also increased. Along with the results of pre-formulation screening, stability tests were conducted to obtain lead stability data. Following the initial stability tests, prototype stability tests were carried out to obtain the optimal formulation for the bulk drug substance and the drug product. The preliminary tests and stability tests are discussed in detail below.

[0188] 1. method A. Exterior Appearance was determined by visual observation using normal laboratory light against both white and black backgrounds.

[0189] B. C5 bond The C5 binding ELISA is an efficacy assay for ALXN1210. The procedure of this test is a direct binding assay using colorimetric detection and was used to verify the ability of ALXN1210 to bind to the human C5 complement protein, which is its target. The Polysorp microtiter plate was coated with human C5 protein and blocked with bovine serum albumin (BSA). The standard curve was generated from the reference material of ALXN1210. The reference material and the test samples were prepared at three dilutions determined to fall within the working range of the assay. After incubation with the standards and samples, the plate was washed and then incubated with mouse anti-human IgG4 conjugated to horseradish peroxidase (HRP). The plate was washed again and then developed using the substrate 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), (ABTS). The amount of reacted substrate was read by spectrophotometric photometry on a plate reader at 405 nm. The absorbance measurements were proportional to the concentration of ALXN1210 bound to C5 on the plate. Four-parameter curve fitting was applied to the standard curve and the results of the assay material and test samples were interpolated from the curve. The results of the test samples were compared to the results of the reference material and the relative activity (%) was reported.

[0190] C. density Density measurements using a DMA 4500 densitometer were determined via the U-tube principle. A hollow U-shaped glass tube was filled with the sample and then electronically excited at the lowest possible amplitude. The density was determined via the following relationship: ρ = A(τ 2 ) - B ρ = density τ = period of vibration

[0191] A and B are instrument constants and were determined via instrument calibration using two substances of known density.

[0192] D. Differential scanning fluorescence Differential scanning fluorescence (SIfluorescence) measures changes in thermal stability by running a thermal denaturation curve in the presence of a fluorescent dye, such as Sypro Orange. When a protein is spread out, the exposed hydrophobic surface binds to the dye, increasing fluorescence and the hydrophobic exposure temperature. h A stability curve is created that has characteristic midpoint values ​​at a given temperature.

[0193] E. Dynamic light scattering method Dynamic light scattering measures the size and interactions of proteins, nanoparticles, and other macromolecules in situ within microwell plates by using an illumination system that allows the wells in the microplate to be imaged using a 3-megapixel onboard camera. Variations in light scattering due to Brownian motion provide a diffusion coefficient, which is related to the hydrodynamic radius of the particles present in the solution.

[0194] F. HPLC gel infiltration method Using gel permeabilization (size exclusion) HPLC, monomeric IgG was identified from larger multimeric antibody species that may arise from monomer aggregation. Test samples were injected onto a TSK gel G3000 SWXL column equilibrated with phosphate-buffered saline, pH 7.0, followed by compositional elution. Protein peaks were monitored at 214 nm, and the purity percentage of monomeric IgG was expressed as the percentage of total integrated peak area. Detection of larger mass multimers was achieved by observing peaks eluting before the monomer peak.

[0195] G. Imaging Capillary Electrophoresis (iCE) This method uses the Protein Simple iCE280 or iCE3 system. This system performs free solution IEF in a capillary column and detects the focused protein zone using a full-column UV detector. Samples were prepared by pre-mixing ALXN1210, a carrier amphoteric electrolyte, and a pI marker. The sample was loaded into a capillary cartridge, and the electrolyte tanks at each end of the capillary were filled with acid and base. A voltage was applied, and the analyte was focused at its pI. A CCD camera captured UV absorbance images of the entire capillary column every 30 seconds, allowing real-time monitoring of the focusing process. The resulting separation patterns were captured and analyzed. The pI of proteins present in the sample was interpolated from the positions of the pI markers mixed into the sample.

[0196] H.Lab on Chip (LoC) This method tests the homogeneity and purity of the product. Non-reduced samples were denatured by treatment with lithium dodecyl sulfate (LDS). Reduced samples were denatured by treatment with lithium dodecyl sulfate (LDS), and the disulfide bonds were broken with dithiothreitol (DTT). Polypeptide chains were mixed with a fluorescent dye, which was then bound to LDS and separated according to molecular size by microcapillary electrophoresis. Proteins were detected and quantified by laser-induced fluorescence.

[0197] I. Osmotic pressure The osmotic pressure of the samples was determined using a freezing point depression osmometer. The osmometer was calibrated before use with commercially available, approved osmotic standards of 50 mOsm / kg and 850 mOsm / kg that encompassed the sample range. A reference 290 mOsm / kg solution was used to confirm successful calibration before validating the samples. Samples were triple-validated, and the average of the sample measurements was reported.

[0198] J.pH pH measurements were performed using a KCl composite electrode without protein-resistant saturated silver, and the associated meter and temperature monitor. The meter was calibrated before use with a commercially available solution in the appropriate pH range (i.e., pH 4.0 to pH 7.0).

[0199] Protein concentration using K.SoloVPE Protein concentration was determined using the path length variable technique in the test sample using the absorbance coefficient of 1.479 determined theoretically using the absorbance at 280 nm. In this method, triple absorbance readings were taken per sample.

[0200] L. Viscosity Viscosity measurements using an AMVn viscometer were determined via the principle of the rolling ball. A hollow tube was filled with the sample and a solid ball of known density, and then tilted at a known angle. The time it took for the ball to move from one side of the tube to the other was determined and used to calculate the viscosity via the following relationship. η = K * (ρb - ρs) * tr η = dynamic viscosity (mPa*s) K = proportionality constant ρb = density of the ball (g / mL) ρs = density of the sample (g / mL) tr = rotation time of the ball

[0201] The sample density determined using a DMA 4500M densimeter was used as ρs to calculate the viscosity.

[0202] Determination of sub-visible particles (MGI) by M. microflow imaging. The objective was to evaluate all insoluble particles in the formulation using microflow imaging (MFI). Samples were taken from storage at 2–8°C and directly validated using MFI with a BOT1 autosampler. The samples were inverted six times, and then loaded into the BOT1 to ensure complete mixing of the particles. The samples were loaded into three consecutive wells, with each well yielding one measurement result from a total of three replicate experiments. Three mixing cycles were incorporated into the BOT1 to further ensure uniform mixing.

[0203] 2. Formulation development Figures 1-5 and Tables 1-5 show the experimental results of the initial formulation development of ALXN1210 at high concentrations.

[0204] In the initial experiment, the addition of an amino acid to ALXN1210 in sodium phosphate buffer was observed to have an effect on its milky appearance. It was determined that the milky appearance was due to a lack of charge in response to the high concentration of antibody molecules in the solution. A series of experiments, as described below, were conducted to optimize the specific amino acid and its concentration necessary to produce a stable, clear solution. Based on these experiments, it was determined that the addition of a positively charged amino acid (L-arginine) reduced the milky appearance of the 50 mg / mL ALXN1210 sample in sodium phosphate buffer. Visual inspection of the vial also reached the same conclusion (data not shown).

[0205] Furthermore, the following experiments evaluated various initial buffering systems to identify a highly concentrated ALXN1210 formulation by concentrating the antibody using a 10 mg / mL ALXN1210 IV formulation and performing buffer exchange. As shown in Table 1 below, all pooled samples obtained the desired pH by performing a final buffer exchange of 1:1000. The concentration range of the pooled samples was 35.3–54.0 mg / mL. The recovery rate % after buffer exchange ranged from 70.6% to 108%. Visual results show that the vials buffered with 25 mM histidine pH 7 and 25 mM phosphate pH 7 were clear and colorless compared to eculizumab, while the solutions in all other buffered vials were milky white. Image-guided capillary electrophoresis (iCE) results showed a pI range of 5.98–6.54, with a major pI range of 6.19–6.24 and an area percentage of 63.1%–65.9%. Size exclusion chromatography (SEC) results showed monomer percentages (purity) of 98.48%–98.98%.

[0206] Table 1: Buffer exchange ALXN1210 10 mg / mL to 50 mg / mL (pooled sets 1-3) [Table 1]

[0207] As shown in Figure 1, salt titration results of a pH 7 histidine buffer exchange sample using DLS indicate that self-binding in ALXN1210 increases as the salt concentration increases.

[0208] As shown in Figure 2, results from L-arginine titration using dynamic light scattering (DLS) indicate that 25 mM L-arginine is the minimum amount required to reduce the milky white color of ALXN1210 at 50 mg / mL.

[0209] As shown in Figure 3, salt titration results of pH 7 phosphate buffer exchange samples using DLS show that the absence of salt and the addition of 150 mM salt resulted in the least self-binding in ALXN1210. Compare peaks 2 and 5 with the labeled peaks.

[0210] As shown in Figure 4, the results from buffer exchange samples using DSF indicate that the hydrophobic pocket is not exposed in ALXN1210. The citrate and acetate buffers at pH 5 and 6 have low thermal stability and the lowest melting point (Tm), while the histidine and phosphate buffer at pH 7 is the most stable and has the highest Tm.

[0211] As shown in the visual results in Table 2, approximately 100 mg / mL of ALXN1210, when 25 mM L-arginine is added to 25 mM phosphate buffer at pH 7, is clear and colorless.

[0212] Table 2: Appearance of ALXN1210 100 mg / mL sample [Table 2]

[0213] As shown in Figure 5, DLS results for ALXN1210 at 10 mg / mL, and ALXN1210 with and without L-arginine at 114 mg / mL, indicate that adding 25 mM L-arginine to at least 100 mg / mL samples brings them closest to the 10 mg / mL reference sample. The at least 100 mg / mL samples without added L-arginine are higher-order species, suggesting self-binding.

[0214] The osmotic pressure results shown in Table 3 indicate that ALXN1210 in 25 mM histidine pH 7.2 containing 8% sucrose or 4.5% sorbitol is within the desirable osmotic pressure range of 275–320. The osmotic pressure of ALXN1210 in 25 mM phosphate supplemented with 7% sucrose or 4% sorbitol, 25 mM L-arginine, pH 7 also falls within the desirable osmotic pressure range.

[0215] Table 3: Osmotic pressure of ALXN1210 in various formulations [Table 3]

[0216] The viscosity results shown in Table 4 indicate that as the concentration of ALXN1210 increases, the viscosity of the histidine and phosphate buffer solutions of ALXN1210 also increases. The density results shown in Table 4 indicate that there is no significant change in the density of the histidine and phosphate buffer solutions when the concentration changes.

[0217] Table 4: Viscosity and density of ALXN1210 samples at various concentrations [Table 4]

[0218] As shown in Table 5, the addition of L-arginine base significantly increased the pH of the sample. Adding L-arginine QS containing a single sodium phosphate base to the sample raised the pH by 1 pH unit. Adding L-arginine HCl lowered the pH by approximately 0.25 pH units. However, the milky appearance decreased with the addition of L-arginine HCl, then L-arginine QS (single sodium phosphate base), and finally L-arginine base.

[0219] Table 5: Effect on pH and appearance of L-arginine buffer ALXN1210 for 25 mM L-arginine addition [Table 5]

[0220] 3.Term Stability Figures 6-21 show the results of the initial stability tests. These results indicate that the histidine formulation was the least stable and the phosphate formulation the most stable after 2 months at 2-8°C, 23-27°C, and 37°C. Furthermore, as revealed by size exclusion chromatography, sorbitol and sucrose were comparable in the phosphate formulation after 2 months at 2-8°C and 23-27°C. However, sorbitol was slightly more stable than sucrose at 37°C after 2 months. Dynamic light scattering results showed that the addition of sucrose or sorbitol did not significantly alter the phosphate sample containing 25 mM L-arginine after 2 months at 2-8°C. The dynamic light scattering results for the histidine sample at T=2 months could not be superimposed due to high dispersion between acquired data. This indicates that the histidine formulation is less stable than the phosphate formulation. The results of the 5-day freeze-thaw cycle showed no significant changes between T=0 and the 5-cycle freeze-thaw cycle.

[0221] 4. Prototype formulation Figures 22-46 show the results of prototype stability testing. These results indicate that all phosphate formulations at 75 mg / mL and 100 mg / mL (bulk drug substance (BDS) and drug product (DP)) remained stable throughout the stability testing at 2–8°C, -20°C, and -80°C. All 100 mg / mL bulk drug substance formulations remained stable after five freeze-thaw cycles at -20°C and -80°C, showing no significant changes.

[0222] 5. Preliminary conclusions before short-term decomposition testing Based on these test results, the optimal formulation for high-concentration ALXN1210 was determined. Preliminary experiments suggested that the addition of L-arginine reduced the milky appearance of 100 mg / mL ALXN1210. Initial stability tests led to the selection of a lead formulation in phosphate buffer containing more than 50 mg / mL of L-arginine. Prototype stability tests determined that the initial optimal formulation of ALXN1210 was 100 mg / mL in 50 mM phosphate buffer, 5% sucrose, 25 mM arginine, and pH 7.4.

[0223] 6. Development of the ultimate optimal formulation The suitability of the initial optimal formulation (100 mg / mL of ALXN1210 in formulation buffer (50 mM sodium phosphate, 25 mM arginine, and 5% sucrose, pH 7.4)) was assessed through a short-term degradation test to evaluate whether polysorbate 80 (PS80) or other surfactants were necessary to prevent degradation. NOF America Corporation reported to contain 0.05% (w / v) oleic acid with a purity of over 99% was also used. The term PS80 refers to two widely used surfactants: POLYSORBATE 80(HX2)(trademark) and AVANTOR(trademark) 4117 and JTBaker(registered trademark) Polysorbate 80, both consisting of a fatty acid blend containing oleic acid and palmitic acid. Both products are often referred to as TWEEN 80(registered trademark) and are nonionic surfactants derived from polyethoxylated sorbitan and oleic acid, containing hydrophilic groups derived from ethylene oxide polymers.

[0224] The test methods used to evaluate the feasibility of JTbaker avantor 4117 PS80 in a 100 mg / mL ALXN1210 formulation are listed in Table 6 below. Details of each test method and a description of the method are provided.

[0225] Table 6: Test methods for the usability of the JTBaker Avantor 4117 PS80 [Table 6]

[0226] Vials containing 100 mg / mL of ALXN1210, either JTBaker avantor 4117 PS80 or HX2 NOF PS80, were visually inspected. All samples, after degradable storage at 45°C and 5 days of additional agitation at 2–8°C, showed no visible particles or any significant color change. The results are shown in Table 7.

[0227] Table 7. Visual inspection of 100 mg / mL ALXN1210 in a 5cc vial containing either J.TBaker avantor 4117 PS80 or HX2 NOF PS80. [Table 7]

[0228] Under different degradation conditions, a slight decrease was observed at a concentration of ALXN1210 mg / mL. Table 8 lists all the concentration measurement results.

[0229] As shown in Figure 2, the ALXN1210 100 mg / mL formulation containing either 0.05% Avantor PS80 or HX2 NOF PS80 showed no significant concentration changes when exposed to the same degradation conditions.

[0230] Table 8: Concentration measurement results [Table 8]

[0231] As shown in Table 8 above, ALXN1210 100 mg / mL formulations containing either 0.05% Avantor PS80 or HX2 NOF PS80 showed no significant concentration changes when exposed to the same degradation conditions. The concentrations of ALXN1210 100 mg / mL formulations containing either PS80 or HX2 NOF PS80 remained comparable.

[0232] Turbidity was measured by monitoring the absorbance at 650 nm. The measurement results are shown in Table 9.

[0233] ALXN1210 100 mg / mL formulations containing either 0.05% Avantor PS80 or 0.05% HX2 NOF PS80 showed no significant turbidity changes. Turbidity remained stable at all time points and under all degradation conditions in this study.

[0234] Table 9: Turbidity was measured by monitoring Abs 650 nm. [Table 9]

[0235] A decrease in monomer percentage was observed in samples incubated at 45°C for 7 or 14 days, followed by additional shaking at 2–8°C (200 RPM). This was predicted to be due to degradation conditions. However, there was no significant difference in monomer percentage between ALXN1210 100 mg / mL formulations containing either 0.05% Avantor PS80 or 0.05% HX2 NOF PS80 when exposed to the same time and conditions.

[0236] The monomer percentage data is shown in Table 10. Figure 4 shows the predicted decrease in monomer percentage after degradation conditions and that there was no significant difference between the ALXN1210 100 mg / mL formulation containing either 0.05% Avantor PS80 or 0.05% HX2 NOF PS80 when exposed to the same time and conditions.

[0237] Table 10: Monomer percentage of samples incubated at 45°C for 7 or 14 days, followed by further shaking at 2–8°C (200 RPM). [Table 10]

[0238] As shown in Table 11, the shift towards acidic species was detected by isoelectric focusing after incubating ALXN1210 with either 0.05% JTBaker Avantor Polysorbate 80 or 0.05% HX2 NOF Polysorbate 80 at 45°C for 7 and 14 days. Additional shaking of the sample did not significantly affect the shift of further major peaks towards acidic species.

[0239] Table 11: Isoelectric focusing electrophoresis using CE-SDS [Table 11]

[0240] Shaking stress tests were performed on initial optimal formulations of ALXN1210 formulation buffer containing 50 mM sodium phosphate, 25 mM arginine, and 5% sucrose, pH 7.4, in the presence and absence of two brands of PS80 at a concentration of 0.05%. Degradation was evaluated using the formation of insoluble microparticles. Samples were shaken at 200 rpm at a temperature of 2–8°C. The time points at which the presence or absence of insoluble microparticle formation was measured were 0, 1, 3, and 5 days. As shown in Table 12, the results indicate that adding 0.05% PS80 to the formulation significantly reduces the formation of insoluble microparticles when high-concentration formulations are exposed to short-term stress, such as shaking at 200 rpm.

[0241] Table 12: The generation of insoluble particles was reduced by the addition of PS80 and filtration. [Table 12]

[0242] 7. Conclusion In conclusion, the optimal subQ formulation for 100 mg / mL of ALXN1210 is a pH 7.4 buffer containing 50 mM sodium phosphate, 25 mM arginine, 5% sucrose, and 0.05% PS80.

[0243] Example 2: A Phase 1 trial to evaluate a single dose of ALXN1210, comparing subcutaneous and intravenous administration in healthy subjects. Phase I trials were conducted to evaluate the safety, tolerability, pharmacokinetic (PK) / pharmacodynamic (PD) effects, and immunogenicity of the antibody BNJ441 (also known as ALXN1210) by comparing subcutaneous (SC) and intravenous (IV) administration in healthy subjects.

[0244] 1. Purpose The primary objectives of this study were (1) to evaluate the safety and tolerability of a single dose of ALXN1210 administered subcutaneously to healthy subjects, compared with ALXN1210 administered intravenously, as assessed by physical examination findings, vital signs measurements, immunogenicity, clinical laboratory analysis, and adverse event (AE) assessment; and (2) to determine the absolute bioavailability of subcutaneously administered ALXN1210.

[0245] A secondary objective was to evaluate the PD effect by comparing subcutaneously administered ALXN1210 with intravenously administered ALXN1210, as assessed by free C5 levels and chicken red blood cell (cRBC) hemolysis.

[0246] 2. Examination Design The overall study design was carried out as shown in Figure 47. This study was a Phase I trial designed to evaluate the safety, tolerability, pharmacokinetics, disease progression, and immunogenicity of a single 400 mg subcutaneous dose of ALXN1210 compared to a single 400 mg intravenously administered dose of ALXN1210 or a subcutaneously administered placebo in 42 healthy subjects. All subjects were screened for eligibility. Subjects who did not meet the eligibility criteria were rescreened for participation in the study if the conditions leading to disqualification were transient, self-limiting, easily treatable, and expected to be resolved at the time of administration.

[0247] Six subjects were initially blindly assigned to Cohort 1a in a 2:1 ratio and received either a single 400 mg subcutaneous dose of ALXN1210 or a single subcutaneous dose of placebo. Clinical safety data for the first 48 hours after administration were evaluated for subjects in Cohort 1a, after which enrollment to Cohort 1b or 2 was initiated. Subsequently, 36 subjects were randomly assigned in a 2:1 ratio to either Cohort 1b (N=24) or Cohort 2 (N=12). Within Cohort 1b, 24 subjects were further blindly assigned in a 5:1 ratio and received either a single 400 mg subcutaneous dose of ALXN1210 (20 subjects) or a single subcutaneous dose of placebo (4 subjects). The 12 subjects in Cohort 2 received a single 400 mg intravenous dose of ALXN1210 in an open-label manner.

[0248] All enrolled subjects were included in the analysis as needed. Subjects from cohorts 1a and 1b were combined into a single cohort for analysis. Subjects participated in the study for up to 39 weeks, including a screening period of up to 70 days and a subsequent 200-day follow-up period for safety, PK, PD, and immunogenicity assessments after administration of the investigational drug.

[0249] Forty-two subjects were evaluated for the primary and secondary objectives of this study. Cohort 1a consisted of 6 subjects (4 receiving subcutaneous ALXN1210 and 2 receiving subcutaneous placebo); Cohort 1b consisted of 24 subjects (20 receiving subcutaneous ALXN1210 and 4 receiving subcutaneous placebo); and Cohort 2 consisted of 12 subjects (ALXN1210 intravenously).

[0250] 3. Rationale for dosage A single dose of 400 mg was equivalent to 4 mL and was administered subcutaneously by four 1 mL injections into the abdomen. A single dose of 400 mg of ALXN1210 SC was predicted to have an acceptable safety profile. The single doses of 400 mg of ALXN1210 SC and placebo SC administered as described in this protocol were expected to provide data that could be used to create simulations of multiple doses, allowing for the planning of the necessary dosing regimens to achieve therapeutic serum concentrations (≥50 μg / mL) in patients.

[0251] Based on a summary of clinical safety data obtained from six subjects in Cohort 1a during the first 48 hours after administration, 36 subjects were randomized in parallel to Cohort 1b and Cohort 2. Enrollment to Cohort 1b and Cohort 2 proceeded as described in Table 13.

[0252] The rules for the toxic population were as follows: Toxicity refers to a clinically significant drug-related adverse reaction. "Cohort progression" refers to progression to a sequential dosing / dosing regimen in accordance with the dosing progression rules and minimum data requirements. "Suspension" refers to the cessation of further IMP administration at the applicable dosing level / dosing regimen and the cessation of further cohort progression. Table 13: Toxicity rules [Table 13-1] [Table 13-2]

[0253] 4. Evaluation Schedule The timing of the test procedures used is shown in Tables 14-15.

[0254] Table 14: Evaluation Schedule: From Screening to First Visit [Table 14-1] [Table 14-2]

[0255] Table 15: Evaluation Schedule: Visits 2-14 [Table 15]

[0256] 5. Selection of targets and withdrawal To be eligible for this examination, candidates must meet all of the following criteria: 1. The subjects must be healthy at the time of administration, and their age must be between 25 and 55 years, including both ends of the age range. 2. The Body Mass Index (BMI) ranges from 18 to 29.9 kg / m², including both extremes. 2 , and weight including both ends is 50-100 kg. 3. The QT interval (QTcF) corrected using the Fridericia formula at screening and before administration on day 1 was ≤450 for men and ≤470 for women. 4. The patient is willing and able to submit written informed consent and adhere to the trial visit schedule. 5. Documentation of vaccination with MCV4 at least 56 days prior to administration and within 3 years thereof is required. The documentation must include positive antibody titers, and an immune response must be confirmed before administration of the investigational drug. 6. Administer serogroup B meningococcal vaccine at least 56 days before the Day 1 dose, administer a booster at least 28 days before the Day 1 dose, and allow at least 28 days between the first and second injections. 7. Women of potential pregnancy who are heterosexually active must use a highly effective or acceptable method of contraception as defined below, starting at screening and continuing for at least 6 months after administration of the study drug. Prophylactic antibiotic administration is required during the study period, and prophylactic administration may impair the effectiveness of hormonal contraception. Therefore, those using hormonal contraception were recommended to also use a barrier contraceptive (e.g., condoms or diaphragms containing spermicide) during the period of prophylactic antibiotic administration. Men who are heterosexually active and who are married to or partnered with a woman of potential pregnancy, or a woman who is pregnant or breastfeeding, must agree to use a barrier contraceptive (male condom) during the treatment period and for at least 6 months after administration of the study drug. Barrier contraception was required even if there was a written medical evaluation of the success of the vasectomy. Female spouses or partners of men of potential pregnancy must use a highly effective method of contraception as defined above, or an acceptable method of contraception as defined below, starting at the time of screening and continuing for at least six months after administration of the investigational drug. Male subjects must not donate sperm during the screening period, the treatment period, and for at least six months after administration of the investigational drug.

[0257] Individuals meeting any of the following exclusion criteria were not eligible to participate in this study: 1. Individuals who are in close, long-term contact (as defined by living under the same roof or providing personal care) with persons younger than 1.2 years or older than 65 years, or who are immunocompromised or have one of the following underlying conditions: anatomical or functional asplenia (including sickle cell anemia); congenital complement deficiency, propardin deficiency, factor D deficiency, or primary antibody deficiency; acquired complement deficiency (e.g., having received eculizumab); or human immunodeficiency virus (HIV). 2. Subjects who meet any of the following criteria: professionals who have been exposed to environments with a high risk of meningococcal disease; researchers, industrial laboratory workers, and clinical laboratory workers who have been routinely exposed to meningococcal bacteria; military personnel undergoing basic training (military personnel may be at increased risk of contracting meningococcal bacteria when placed in urban warfare situations); daycare center workers; university students or those living on university campuses; and those who plan to travel to or have traveled within the six months prior to administration to areas where meningococcal meningitis is endemic during the course of this study (e.g., India, sub-Saharan Africa, or the Hajj pilgrimage to Saudi Arabia). 3. A history of any Neisseria infection. 4. A history of recurrent infections of unknown cause, or an infection requiring systemic antibiotic treatment within 90 days prior to administration. 5. HIV infection (provided by HIV-1 or HIV-2 antibody titer). 6. Acute or chronic hepatitis B virus (HBV) infection. All subjects were required to undergo hepatitis B surface antigen (HBsAg) testing prior to enrollment. Subjects positive for HBsAg were not enrolled. For subjects negative for HBsAg, the following testing algorithm was required: If hepatitis B core antibody (HBcAb) was negative, the subject was eligible for enrollment. If HBcAb was positive, hepatitis B surface antibody (HBsAb) was tested. If both HBcAb and HBsAb were positive, the subject was eligible for enrollment. If HBcAb was positive and HBsAb was negative, the subject was not enrolled. 7. Acute or chronic hepatitis C virus (HCV) infection (provided by antibody titer). 8. Acute systemic viral or fungal infection within 14 days prior to administration. 9. A positive or inconclusive result from the QuantiFERON®-TB test suggests a possible tuberculosis (TB) infection. 10. A history of latent or active TB, or exposure to an endemic area within 8 weeks prior to the screening visit. 11. The study included women who were breastfeeding, or women who were heterosexual, not willing to use contraception, and were premenopausal. Menopause was defined as having no other cause, having had amenorrhea for 12 consecutive months or more, and having recorded serum follicle-stimulating hormone levels of ≥40 mlU / mL and estradiol concentrations of ≤110 pmol / L within 6 months prior to administration of the study drug. 12. A positive serum pregnancy test is performed at the time of screening or on day -1. 13. Serum creatinine is elevated above the upper limit of normal (ULN) of the laboratory reference range at the time of screening or on day 1. 14. Alanine aminotransferase (ALT) or aspartate aminotransferase (AST) levels are higher than the laboratory reference range ULN at screening, or more than 1.5 times the laboratory reference range ULN on day -1. 15. Any of the following blood test results: At screening or on day 1, hemoglobin <130 g / L for men and <115 g / L for women, hematocrit <0.37 L / L for men and <0.33 L / L for women, or white blood cell (WBC) count <3.0 x 10 3 / μL, absolute neutrophil count <2.0x10 3 / μL, and platelet count <150 or >400x10 3 / μL. - Clinical laboratory results of a complete blood cell count (CBC) deemed clinically relevant and unacceptable by the principal investigator on day 1. 16. A history of complement deficiency, or complement activity below the normal range as evaluated by CAP ELISA at the time of screening. 17. History of malignant tumors. However, this excludes non-melanoma skin cancer or cervical carcinoma in situ that has been treated and for which there is no evidence of recurrence. 18. Participate in a clinical trial within 30 days prior to the start of administration on day 1, or use either experimental small molecule therapy within 30 days of administration on day 1. 19. Participation in multiple mAb clinical trials, or participation in an mAb clinical trial within 12 months prior to screening, during which the subject was exposed to the active investigational drug. Subjects who participated in only one mAb trial could be considered enrolled if that trial was completed before 12 months prior to screening. 20. Before exposure to ALXN1210. 21. Major surgery or hospitalization within 90 days prior to administration. 22. History of allergy to excipients of ALXN1210 (e.g., polysorbate 80). 23. Record of allergy history to penicillin or cephalosporins. 24. A history of a serious allergic reaction (e.g., anaphylaxis or angioedema) to any product (food, medicine, etc.). 25. Currently smokes more than 10 cigarettes per day (past smoking may be permitted at the discretion of the principal investigator). 26. A history of illegal drug addiction or severe alcoholism within one year prior to the screening visit. Or, regular alcohol use within six months prior to the screening visit (more than 14 units of alcohol per week [1 unit = 150 mL of wine, 360 mL of beer, or 45 mL of 40% alcohol]). 27. Positive urine screening test for drug intoxication at the time of screening or on day 1. 28. Alcohol consumption within 48 hours prior to administration of the test drug, or a positive alcohol breath test on day 1. 29. Plasma donation within 7 days prior to administration. Blood donation or loss of more than 50 mL within 30 days prior to administration, or more than 499 mL within 56 days prior to administration (excluding the amount collected during screening). A history of continuous use of topical, inhaled, or systemic steroids for more than 30.28 days, or a history of any inhaled or topical immunosuppressive therapy within 90 days prior to administration of the investigational drug. 31. Use of prescription medications (excluding oral contraceptives) within 14 days prior to administration of the investigational drug, unless prior approval has been obtained from the sponsor. 32. Regular use of non-prescription or over-the-counter (OTC) medications, including herbal medicines and supplements, within 14 days prior to administration of the investigational drug. Multivitamins, acetaminophen ≤2 g per day, and topical skin products that do not involve significant systemic absorption are permitted. 33. A clinical diagnosis of any autoimmune or rheumatic disease (e.g., systemic lupus erythematosus, rheumatoid arthritis). 34. Vaccination with a live attenuated vaccine 28 days prior to administration, or planned vaccination during the course of the study (excluding vaccinations planned in the study protocol). Vaccination with inactivated or recombinant influenza vaccines was permitted. 35. The patient had a fever (a temperature exceeding 37.6°C) within 14 days prior to administration (e.g., fever associated with symptomatic viral or bacterial infection). 36. Any medical history, condition, or risk that the principal investigator has indicated could interfere with the subject's full participation in this study, or with compliance with the protocol, or that could pose any additional risk to the subject, or that could complicate the evaluation of the subject or the study results.

[0258] 6. Infectious disease To mitigate the risk of infection associated with terminal complement inhibition, the subjects in this study received the following: 1. MCV4 vaccination must be administered at least 56 days prior to the ALXN1210 administration on day 1 (if MCV4 vaccination has not been received within the last 3 years, or if the subject has already been vaccinated but there is no appropriate documentation to prove past vaccination). 2. Two injections of serogroup B meningococcal vaccine. The first injection must be administered at least 56 days before the first day's dose, and the booster injection must be administered at least 28 days before the first day's dose, with at least 28 days between the first and second injections. 3. Prophylactic antibiotic treatment with oral penicillin V 500 mg twice daily (1 x 10) until complement activity normalizes (as determined by the CH50 assay). 6 (Equivalent to a unit). The first dose of the antibiotic was administered orally on the evening of day 1, prior to the administration of the day 1 dose of the investigational drug. For the outpatient portion of this study, participants were instructed to take the antibiotic at approximately the same time each scheduled day (twice daily). Participants were monitored daily to ensure they were adhering to the antibiotic prophylactic regimen using an appropriate system (e.g., written messages).

[0259] The following findings support the prophylactic administration of antibiotics in this single-dose study: 1. Penicillin was the preferred drug for the elimination of meningococcal bacteria in carriers. 2. Complement-deficient patients who received monthly injections of benzathine penicillin G as prophylactic treatment for recurrent meningococcal disease over a period of 2.2 to 4 years had significantly fewer Neisseria infection symptoms than complement-deficient patients who did not receive prophylactic treatment (Figueroa JE, et al., Clin. Microbiol. Rev. 1991 Jul;4(3):359-95). 3. High levels of penicillin resistance induced by plasmid-encoded β-lactamase were rare in meningococcal strains (Yazdankhah SP, et al., J. Med. Microbiol. 2004 Sep;53(Pt 9):821-32). 4. Prophylactic antibiotic administration of 500 mg of penicillin V orally twice daily was provided by some physicians in the treatment of patients with PNH and aHUS using eculizumab (Kelly RJ, et al., Blood 2011 Jun 23;117(25):6786-92 and Leeds Teaching Hospitals NHS Trust, Kings College Hospital NHS Foundation Trust. National Specialised Commissioning Team (NSCT) Service Specification Paroxysmal Nocturnal Haemoglobinuria (PNH). 2013). 5. Uncertainty regarding the effectiveness of vaccines in immunocompromised patients has been raised in several countries, such as France, and prophylactic antibiotic administration during eculizumab treatment is recommended for patients with PNH and aHUS (Zuber J, Fakhouri F, Roumenina LT, Loirat C, Fremeaux-Bacchi V. Use of eculizumab for atypical haemolytic uraemic syndrome and C3 glomerulopathies. Nat. Rev. Nephrol. 2012 Nov;8(11):643-57).

[0260] 7. Past and concomitant medications and medical procedures Previous medications (any medications or substances taken by the subject from within 14 days prior to the subject signing the ICF until the administration of the investigational drug) and concomitant medications (any medications or substances taken by the subject from after the administration of the investigational drug until the completion of the final study visit) were recorded in the subject's electronic case report form (eCRF). Previous medical procedures (any therapeutic interventions performed from within 14 days prior to the subject signing informed consent until the administration of the investigational drug [e.g., surgery / biopsy, physiotherapy]) and concomitant medical procedures (any therapeutic interventions performed after the administration of the investigational drug until the completion of the final study visit [e.g., surgery / biopsy, physiotherapy]) were recorded in the subject's eCRF.

[0261] Adjunctive therapy included any medications or substances administered to the subject from the time of screening for the study until the completion of the final study visit. During the study period, subjects were instructed not to start taking any new medications, including non-prescription drugs and herbal preparations, without permission from the principal investigator. The occasional use of over-the-counter (OTC) antipyretics or analgesics (e.g., acetaminophen) was permitted during the study period.

[0262] Concomitant medical procedures were defined as any therapeutic intervention (e.g., surgery / biopsy, physiotherapy) or non-experimental diagnostic assessment (e.g., blood gas measurement, bacterial culture) performed between the time the subject signed informed consent and the final trial visit. Concomitant medical procedures were not permitted unless medically justified.

[0263] 8. Randomization and blinding Eligible patients who met the study inclusion and exclusion criteria were assigned a unique number for enrollment and randomization. This study was a partially blinded study, as follows: • Cohort 1a. The administration (single dose of 400 mg of ALXN1210 SC or placebo SC) was double-blind. Participants in Cohort 1a were randomly assigned in a 2:1 ratio (4 received ALXN1210 SC, 2 received placebo SC, N=6). • Cohort 1b. The administration (single dose of 400 mg of ALXN1210 SC or placebo SC) was double-blind. Participants in Cohort 1b were randomly assigned in a 5:1 ratio (20 received ALXN1210 SC, 4 received placebo SC, N=24). • Cohort 2 administration (single dose of 400 mg of ALXN1210 IV) was open-label (N=12). During administration in Cohort 2, both the subjects and the on-site medical / nursing staff were aware of the medication / dosage being administered. During administration in cohorts 1a and 1b, on-site medical / nursing staff at the study centers and in the subjects were blinded to the allocation of the investigational drug. Pharmacy staff preparing the SC injections were not blinded, nor were those administering the investigational drug. On the other hand, all other study center staff involved in the safety assessment remained blinded to the allocation of the investigational drug. Sponsor staff were deblinded as needed (e.g., to monitor that the SC injections were properly prepared, to determine the reporting requirements for SAEs), but any information regarding the allocation of the investigational drug was not shared with study center staff.

[0264] 9. Description of the test drug The products surveyed are listed in Table 16.

[0265] Table 16: Products surveyed [Table 16]

[0266] 10. ALXN1210 and placebo Each vial of ALXN1210 SC contained 100 mg of ALXN1210 (100 mg / mL) in 50 mM sodium phosphate, 25 mM arginine, 5% sucrose, and 0.05% polysorbate 80. ALXN1210 SC was formulated at pH 7.4 and supplied in 2 mL single-use vials as a fully formulated, sterile, preservative-free 100 mg / mL aqueous solution of ALXN1210. Each vial of ALXN1210 SC contained a slight overfill to ensure that 1 mL (100 mg of ALXN1210) could be reliably drawn for administration.

[0267] Each dose of placebo SC contained 0.9% sodium chloride injection, pH Eur, or BP, and was the same as the dose specified for cohorts 1a and 1b.

[0268] Each vial of ALXN1210 IV contains 150 mg of ALXN1210 in 10 mM sodium phosphate, 150 mM sodium chloride, 0.02% polysorbate 80, and water for injection. ALXN1210 IV was formulated at pH 7.0 and supplied in 20 mL single-use vials as a fully formulated, sterile, preservative-free 10 mg / mL aqueous solution of ALXN1210. ALXN1210 IV was diluted with 0.9% sodium chloride injection, Ph Eur, or BP and administered by intravenous infusion at a maximum rate of 333 mL / hour, except for interruptions for safety and technical reasons.

[0269] The ALXN1210 vials were stored under refrigerated conditions at 2°C to 8°C (36°F to 46°F) and protected from light. The ALXN1210 vials were not frozen or shaken. ALXN1210 SC and placebo SC were prepared in a blind manner in syringes for SC administration. Neither ALXN1210 SC nor placebo SC were diluted. ALXN1210 SC and placebo SC were placed directly into syringes.

[0270] ALXN1210 IV is a commercially available saline solution (0.9% sodium chloride injection, pH Diluted to Eur or BP, and designed for IV infusion at a maximum rate of 333 mL / hour, except for interruptions for safety and technical reasons.

[0271] ALXN1210 IV was diluted with 0.9% sodium chloride injection, Ph Eur, or BP and then administered (administration solution). The storage period for the administration solution before use was 4 hours at room temperature (15°C to 25°C, 59°F to 77°F). The expiration date and remaining time of the administration solution were calculated from the time the first vial was opened. The dose was administered within the expiration date and remaining time. Each 1 mL syringe of either ALXN1210 SC or placebo SC (4 syringes per subject) was administered within 1 hour of being drawn from the vial into the syringe.

[0272] 11. Administration The total dose of ALXN1210 SC or placebo SC was administered by four 1 mL injections of SC each in the abdomen (Table 17). All four 1 mL injections were administered over 15 minutes. A minimum of 15 minutes had to be allowed between the completion of an injection in one subject and the start of an injection in the next subject.

[0273] Table 17: Dosage criteria chart for the preparation of ALXN1210 SC and placebo SC [Table 17]

[0274] The entire dose of ALXN1210 IV was administered by intravenous infusion at a maximum rate of 333 mL / hour using an IV set equipped with an in-line filter, except for interruptions for safety or technical reasons. A minimum of 15 minutes had to be allowed between the completion of infusion in one subject and the start of infusion in the next subject.

[0275] Table 18: Dosage criteria chart for the preparation of ALXN1210 IV [Table 18]

[0276] 12. Management of potential adverse events during administration of the investigational drug Some subjects treated with IV infusions of monoclonal antibodies experienced synchronous infusion-related reactions accompanied by signs or symptoms that could be classified as acute allergic reactions / hypersensitivity reactions or cytokine release syndrome.

[0277] Participants were carefully monitored during and after administration of the study drug for any symptoms of anaphylaxis or other hypersensitivity reactions, including changes or cessation of the circulatory and / or respiratory systems, or signs of urticaria, arthralgia, myalgia, or other related reaction signs. Appropriate treatment was immediately available. Infusion-related adverse events may occur, and depending on their type and severity, discontinuation of the infusion may be required. Participants were informed of early symptoms and signs of hypersensitivity reactions, including urticaria, swelling of the face, eyelids, lips, or tongue, or dyspnea. Infusion-related reactions were managed using an acute infusion reaction algorithm. Periodic assessments were performed to monitor infusion and infusion site reactions in this study. At least 15 minutes were allowed between the completion of an infusion / injection in one participant and the start of an infusion / injection in the next participant to ensure prompt action in case of a reaction. No more than 6 participants were administered per day. All reactions were treated and considered for continued / escalated administration and toxicity rules. In the event of an anaphylactic reaction, we followed the latest UK Treatment Guideline for Anaphylactic Reactions from the UK Resuscitation Council.

[0278] 13. Pharmacokinetic (PK) and pharmacodynamic (PD) evaluation Following administration of the test drug, serum samples were collected at the following time points for determining serum ALXN1210 concentration and for analysis of total C5 and free C5 concentrations, cRBC hemolysis, and other measures of potential C5 activation. The actual date and time of serum sample collection were recorded and used for PK and PD calculations: • Serum concentrations of ALXN1210 were analyzed from the following sample collection points: before administration (within 15 minutes before the start of infusion / injection [SOI]); at the end of infusion / injection (EOI) on day 1, 30 minutes after EOI, and at the following points after SOI: 2 hours, 4 hours, and 8 hours; day 2 (24 hours); day 3 (48 hours); day 5 (96 hours); day 8 (168 hours); day 15 (336 hours); day 22 (504 hours); day 29 (672 hours); day 36 (840 hours); day 43 (1008 hours); day 50 (1176 hours); day 57 (1344 hours); day 71 (1680 hours); day 90 (2136 hours); day 120 (2856 hours); day 150 (3576 hours); and day 200 (4776 hours). All subjects who provided an appropriate number of serum PK samples and whose concentration-time profiles were characterized were included in the PK analysis population. All subjects who provided PD samples were included in the PD analysis population.

[0279] 14. Evaluation of immunogenicity Serum samples were collected at the following time points: before administration (within 15 minutes before SOI) and on days 15 (336 hours), 29 (672 hours), 57 (1344 hours), 90 (2136 hours), 120 (2856 hours), 150 (3576 hours), and 200 (4776 hours). They were then analyzed for ADA against ALXN1210. Further characterization of antibody responses was performed as needed, based on PK / PD data and safety data for ALXN1210.

[0280] All patients who provided pre- and post-administration samples for ADA were included in the immunogenicity analysis population.

[0281] The immunogenicity assay evaluates the presence of antidrug antibodies (ADAs) against ALXN1210. Detailed instructions for collecting, processing, storing, and transporting serum samples for immunogenicity analysis are provided in the clinical laboratory manual.

[0282] 15. Safety Evaluation Safety assessments included TB testing, physical examination, vital sign measurement, immunogenicity (ADA) testing, clinical laboratory evaluation, ECG, infusion and injection site evaluation (e.g., bleeding, bruising, erythema, swelling, induration, and pain), and monitoring of adverse events. Adverse events were graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events v4.03 (CTCAE v4.03), published on June 14, 2010. Clinical laboratory evaluations included hematological tests, chemistry tests, and coagulation panels; CBC with differential diagnosis; urinalysis; and serum pregnancy tests for female subjects.

[0283] The safety of ALXN1210 was evaluated through clinical and laboratory assessments. The timing of the evaluations is described in the evaluation schedule. Abnormal results were followed up until they resolved or stabilized.

[0284] A review of demographic parameters, including age, sex, race, and ethnicity, was conducted as outlined in the assessment schedule. A complete medical history was collected.

[0285] Vital signs were measured for at least 5 minutes after the subject rested in a supine or semi-prone position and included body temperature (°C, oral), respiratory rate, supine blood pressure, and pulse. The timing of vital sign measurements was specified in the evaluation schedule. Blood pressure or pulse measurements outside the normal range were repeated at the discretion of the principal investigator. All confirmed clinically significant vital sign measurements were recorded as adverse events.

[0286] Weight, height, and BMI were recorded as they appeared on the evaluation schedule. Each examination included the following assessments: overall appearance; skin; head, ears, eyes, nose, and throat; neck; lymph nodes; chest; heart; abdominal cavity; limbs; central nervous system; and musculoskeletal system.

[0287] A triple 12-read ECG was obtained after each subject had rested for at least 5 minutes. The timing of the ECG was described in the evaluation schedule. In addition, continuous electrocardiogram registration was performed at each administration time: from before IV infusion to completion in Cohort 2, and from before SC injection to 3 hours after completion in Cohorts 1a and 1b. Heart rate, PR, QRS, RR, and QT were measured, and the corrected QTcF interval was calculated.

[0288] Blood samples for blood tests, clinical chemistry tests, coagulation tests, and viral serological tests, as well as urine samples for urine tests, urine chemistry tests, and drug and alcohol screenings, were collected as outlined in the evaluation schedule.

[0289] Blood samples were analyzed for the following blood parameters: platelet count, red blood cell (RBC) count, and white blood cell count; automated differentiation (neutrophils, lymphocytes, monocytes, eosinophils, basophils); hemoglobin; hematocrit; and RBC index (mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration). The timing of the blood assessments is described in the assessment schedule.

[0290] Blood samples were analyzed for the following clinical chemistry parameters: blood urea nitrogen, creatinine, glucose; sodium; phosphorus; potassium; chloride; total carbon dioxide; total calcium; magnesium; AST; ALT; gamma-glutamyltransferase; alkaline phosphatase; lactate dehydrogenase; total bilirubin, direct bilirubin and indirect bilirubin; uric acid; albumin; and total protein. Considering that indirect bilirubin is calculated from the total bilirubin and direct bilirubin values, if the direct bilirubin value falls below the limit of quantification, the indirect bilirubin result cannot be obtained.

[0291] Serum follicle-stimulating hormone levels and estradiol concentrations were measured during screening for postmenopausal women to confirm their menopausal status.

[0292] The timing of the chemical evaluation is specified in the evaluation schedule.

[0293] Blood samples were analyzed for prothrombin time, international normalized ratio, and partial thromboplastin time. The timing of coagulation assessments is described in the assessment schedule.

[0294] The urinalysis included specific gravity, pH, glucose, protein, blood, and ketones. Microscopic examination of the urine sample was performed only for abnormal findings. The urine sample was further sent to the pathology laboratory for measurement of protein and creatinine, and the urinary protein:creatinine ratio was calculated. The timing of the urinalysis and urine chemistry evaluation is described in the evaluation schedule.

[0295] Blood samples collected during screening were analyzed for antibody titers for HIV-1, HIV-2, HBsAg, and HCV. All subjects prior to enrollment were required to undergo hepatitis B surface antigen testing. Subjects positive for HBsAg were not enrolled. For subjects negative for HBsAg, the following testing algorithm was required: 1. If the HBcAb test was negative, the subject was deemed eligible for registration. 2. If HBcAb was positive, hepatitis B surface antibody (HBsAb) was tested. a. If both HBcAb and HBsAb were positive, the subject was considered eligible for registration. b. If HBcAb was positive and HBsAb was negative, the subject was not enrolled.

[0296] Urine samples for drug screening were analyzed for the following compounds: amphetamine, barbiturates, benzodiazepines, cocaine, methadone, opium, phencyclidine, methamphetamine, 3,4-methylenedioxy-methamphetamine, and tetrahydrocannabinol (cannabinoid). Alcohol breath testing was performed. If positive before administration, the patient did not proceed to administration. The timing of urine drug testing and alcohol breath testing is described in the evaluation schedule.

[0297] All women underwent pregnancy testing (beta-human chorionic gonadotropin). The timing of the pregnancy tests is indicated in the evaluation schedule.

[0298] Serum samples for the QuantiFERON-TB test were collected as described in the evaluation schedule.

[0299] Appropriate assays to determine complement activity, such as CAP ELISA / C5 (hemolysis) inhibition, were performed during screening to confirm that subjects did not have complement deficiency. Subjects found to have complement deficiency were excluded from participation in this study.

[0300] Serum samples were collected at baseline and during the follow-up period for measurement of CH50 activity using in vitro LIA, and normalization of complement activity was confirmed. If normal CH50 results were obtained from the first CH50 sample of a subject collected during the follow-up period, prophylactic antibiotic administration may be discontinued, and the planned second CH50 sample was not mandatory. If the first and second CH50 samples were not normal, the baseline sample may be analyzed, and further CH50 samples were collected until complement activity recovered.

[0301] Serum bactericidal antibodies (SBA) against meningococcal serogroups A, C, W135, and Y The titer of bactericidal antibody was determined during screening. Subjects showing no immune response were excluded from treatment using the titer measurements.

[0302] Site evaluation was performed for subcutaneous injections or IV infusions. Pain at the SC injection site or IV infusion site was assessed using a Visual Analog Scale (0-10). Pain was not assessed before administration. Induration or reaction measuring 1 cm in size was not reported as an adverse event unless it persisted for more than 24 hours.

[0303] Serum samples were analyzed for anti-drug antibodies (ADAs). The timing of ADA serum sample collection is described in the evaluation schedule.

[0304] 16. Management of Adverse Events The principal investigator is responsible for detecting, evaluating, documenting, and reporting all adverse events (AEs). All AEs were recorded from the time of signing the informed consent form until the completion of the trial. No time limit was set for SAEs considered to be causally related.

[0305] Regardless of causality, all observed or spontaneous adverse events were reported and recorded in the data acquisition system. Adverse events reported by subjects and / or parents or legal guardians, as well as adverse events identified in response to open-ended questions from study personnel, or adverse events revealed by observation, physical examination, or other study procedures, were collected and recorded.

[0306] An AE was defined as any undesirable, unintended sign (e.g., an abnormal laboratory finding), symptom, or illness that occurred during the course of a clinical trial and was temporally related to the use of a drug or medical practice, regardless of whether it was considered to be related to the drug or medical practice.

[0307] Exacerbations of a chronic or intermittent pre-existing condition, including either an increase in the frequency and / or intensity of that condition, were all considered adverse events (AEs).

[0308] Abnormal laboratory findings were considered adverse events (AEs). When abnormal laboratory values ​​were identified, the principal investigator was strongly encouraged to report the diagnosis or sign or symptom rather than the isolated abnormal laboratory value. Abnormal laboratory findings were documented as AEs if any of the following conditions were met: associated with a sign or symptom; required additional diagnostic testing (repeated testing was not considered additional testing); required medical or surgical intervention; deviated from protocol-specified dosage, altered study dosage, or interrupted study; required significant additional treatment; or did not meet any of the above conditions.

[0309] This definition also includes signs or symptoms resulting from: drug overdose, drug withdrawal symptoms, drug abuse, drug interactions, spills, exposure during pregnancy, exposure through breast milk, medication errors, and occupational exposure.

[0310] AE does not necessarily include the following: • Medical or surgical interventions (e.g., surgery, endoscopy, tooth extraction, blood transfusion); conditions leading to such medical procedures were considered AEs (for example, laparoscopic cholecystectomy was a medical procedure or treatment for SAEs of necrotizing gallbladder). • Pre-existing diseases or conditions that were present or detected prior to the screening assessment and that do not worsen. • No adverse medical events have occurred (e.g., hospitalization for scheduled surgery, if it was planned before the start of this study; hospitalization for social and / or convenience reasons).

[0311] Any AE that met any one of the following criteria was recorded as a SAE: SAEs were recorded as any of the following adverse medical events at all doses: 1. To cause death 2. Life-threatening a 3. Hospitalization or extension of hospitalization is required. b Hospitalization does not necessarily include the following: Rehabilitation / Hospice / Nursing Facility • Emergency visit within 24 hours • Scheduled or planned hospitalization / surgery / day surgery • Hospitalization as specified in the protocol • Hospitalization for a pre-existing condition not related to a new AE or an exacerbation of an existing AE 4. Causes a persistent or significant impairment / disqualification. 5. It is a congenital / birth abnormality. 6. It is an important medical event. c a In the definition of "serious," the term "life-threatening" refers to an event in which the subject is at risk of death at the time of the event; it does not refer to an event that could potentially lead to death if it were more severe. b Hospitalization requires the patient to be admitted to the hospital or to have their existing hospitalization period extended. AEs associated with hospitalization or extension of hospitalization were considered SAEs. c In other situations, such as important medical events that are not immediately life-threatening, do not result in death, or do not require hospitalization, but endanger the subject or would require intervention to prevent one of the other outcomes listed above, medical and scientific judgment must be made to determine whether rapid reporting is appropriate. These are also generally considered to be serious. Examples of such events include emergency department or home-based intensive care for allergic bronchospasm; hematopoietic dysfunction or seizures that do not require hospitalization; or the onset of drug dependence or abuse.

[0312] Severity and criticality are to be distinguished. Severity describes the intensity of the AE, while criticality refers to an AE that meets the SAE criteria, as mentioned above.

[0313] All AEs were graded according to the following criteria in CTCAE v4.03, which was released on June 14, 2010. • Grade 1: Mild (signs or symptoms are recognized but are easily tolerated) • Grade 2: Moderate (Discomfort sufficient to interfere with normal activities) • Grade 3: Severe (loss of physical function to the extent that normal activities cannot be performed) • Grade 4: Life-threatening Grade 5: Fatal Changes in the severity of adverse events (AEs) were documented, allowing for the assessment of AE duration at each intensity level being evaluated. Adverse events characterized as intermittent required documentation of occurrence and duration for each symptom if the severity of the intermittent event changed.

[0314] For all AEs (both serious and non-serious), the principal investigator submitted a causal assessment. This assessment was recorded in the data capture system as needed and in any additional format. The definition of the causal assessment was as follows: • Unrelated: This relationship suggests that there was no correlation between the test product and the reported event. • Low probability of correlation: This relationship suggests that while the clinical presentation closely matched causes other than the investigational product, the contribution cannot be ruled out with absolute certainty, and a relationship between the investigational product and adverse events (AEs) cannot be completely ruled out. • Likely related: This relationship suggests that treatment with the test product may have caused or contributed to the adverse event (AE). That is, the event followed a reasonable timeline from the administration of the test drug and / or followed a known reaction pattern to the test product, but it could also have been caused by other factors. • Likely related: This relationship suggests a reasonable timeline between the administration of the investigational product and the event, and proposes a possible link between the event and the investigational product. This is based on the known pharmacological effects of the investigational product, known or previously reported adverse reactions to the investigational product or drug group, or the clinical experience of the principal investigator. • Fully related: Temporal relevance to the test product. Other conditions (concurrent illness, concurrent drug response, or progression / onset of disease) do not appear to explain the event. The event is consistent with the known pharmaceutical profile, improvement after discontinuation, and recurrence upon readmission.

[0315] If a subject experienced a fatal SAE, the following procedure was followed: Fatal SAEs were documented as death / lethal, with the end date being the date of death. If any additional AEs / SAEs were ongoing at the time of death, these events were documented as ongoing without an end date. Unless otherwise stated in the autopsy report or by the principal investigator, there was only one event resulting in death / lethal.

[0316] 17. Statistics Prior to the database locking, a formal statistical analysis plan (SAP) was created and finalized. The safety population consisted of all subjects who received at least one dose of the investigational drug. This population was used for the safety analysis.

[0317] The PK population consisted of all subjects with sufficient serum concentration data to enable the calculation of PK parameters. The PK population was used for summarizing PK.

[0318] The PD population consisted of all subjects with sufficient total C5 concentration data, free C5 concentration data, and cRBC hemolysis data. The PD population was used for summarizing PD.

[0319] The immunogenicity analysis population consists of all subjects for whom pre- and post-administration ADA samples have been collected.

[0320] The total evaluable sample size was 36 subjects, 24 of whom received ALXN1210 SC in Cohort 1 and 12 of whom received ALXN1210 IV in Cohort 2. This provided >80% power to infer that the lower limit of the 90% confidence interval for the ratio of bioavailability between ALXN1210 SC and IV was >0.4, assuming an absolute bioavailability of 0.6 and a variability factor of 0.35. In addition, 6 subjects received placebo SC: 2 in Cohort 1a and 4 in Cohort 1b. Randomization for Cohort 1a was conducted in a 2:1 ratio, and for Cohort 1b in a 5:1 ratio, with subjects receiving either ALXN1210 SC or placebo SC. This resulted in a total of N=42 subjects.

[0321] Generally, descriptive statistics for continuous variables include the number of non-missing values, arithmetic mean, standard deviation, median, minimum, and maximum. Descriptive statistics for PK parameters include the number of observations, arithmetic mean, standard deviation, arithmetic coefficient of variation (%CV), median, minimum, maximum, geometric mean, and geometric %CV. Categorical variables were summarized by cohort and time point using percentages and frequency counts.

[0322] All subjects were included in the summary of the trial continuation status. This summary outlines the frequency and percentage of subjects who were screened, treated, and completed or discontinued the trial, along with the reasons for discontinuation, for each cohort. Demographic and baseline characteristics were summarized for all subjects, both for each cohort and overall.

[0323] Safety analyses were conducted on the safety population and reported for each cohort. Safety analyses included analysis of all AEs, ECGs, laboratory data, physical examinations, and vital sign measurements, presented using descriptive statistics. Inferential statistical analysis was not planned for the safety parameters of this study. The incidence of AEs and SAEs under investigational drug administration was summarized by organ-specific major categories and basic terms for each cohort and overall, based on their association with the investigational drug. AEs under investigational drug administration were also summarized by severity, both by cohort and overall. Serious AEs resulting in withdrawal from this study were listed. Subjects with multiple AEs within a category (e.g., overall, organ-specific major categories, basic terms) were counted as one event within that category. For the severity tables, the most serious event for subjects within a category was counted.

[0324] Changes from normal values ​​in vital sign measurements and clinical laboratory assessments (e.g., chemistry, CBC with differential diagnosis, and urinalysis) were summarized for each cohort. Laboratory parameter values ​​were graded according to the CTCAE. Shift schedules were created for each cohort for these laboratory parameters. These schedules summarize the number of subjects in each normal grade compared to the reference range, and the change to the worst grade assessed during the study and after administration.

[0325] At designated time points, ECG parameters including heart rate, PR, RR, QRS, QT, and corrected QTcF interval were measured. The mean of the triple ECG measurements at the time of collection was calculated, and the change from pre-treatment baseline was evaluated for each cohort.

[0326] An outlier analysis was conducted to summarize the frequency and percentage of subjects meeting any of the following outlier criteria at each hospital visit, for each cohort. • QT, QTcF interval > 450 milliseconds • QT, QTcF interval > 480 milliseconds • QT, QTcF interval > 500 milliseconds • QT and QTcF intervals increase by more than 30 milliseconds from the baseline. • QT and QTcF intervals increase by more than 60 milliseconds from the baseline.

[0327] All concomitant medications were coded using the WHO Drug Dictionary, and their frequency and percentages were summarized.

[0328] Individual serum concentration data for ALXN1210-treated patients, along with the actual sampling date and time, were used to derive PK parameters using a non-compartmental analysis method with Phoenix WinNonlin 6.3 or higher.

[0329] The following PK parameters were derived: Observed maximum blood concentration (C max ), time to observed maximum blood concentration (T max ), the area under the curve (AUC) between serum concentration and time from zero to the last quantifiable concentration. t ), Area under the curve from zero to infinity (AUC ∞ ), terminal phase excretion rate constant (λ Z ), terminal phase excretion half-life (T 1 / 2 ), total clearance (CL or CL / F) and distribution amount (V d or V d / F).

[0330] The geometric mean ratio (ALXN1210 SC / ALXN1210 IV) and its 90% CI are C max AUC t and AUC ∞ The results were computer-calculated and listed. The CI (Continuous Index) was computer-calculated using the variance between subjects. Concentration assessments over time were presented.

[0331] The PD effect of ALXN1210 SC and IV was assessed by evaluating changes in serum total C5 concentration, serum free C5 concentration, cRBC hemolysis, and other measures related to C5 activation over time. Analysis was performed on samples collected as described in the evaluation schedule.

[0332] Immunogenicity measured by ADA was summarized in a tabular format by cohort and by subject.

[0333] Example 3: Results of a Phase 1 trial to evaluate a single dose of ALXN1210, comparing subcutaneous and intravenous administration in healthy subjects. The following is a summary of data from a single-dose Phase I trial, which was substantially conducted as described in Example 2. Specifically, this trial was designed to evaluate the safety, tolerability, pharmacokinetics (PK), pharmacodynamics (PD), and immunogenicity of a single 400 mg subcutaneous dose of ALXN1210 compared to a single 400 mg intravenously administered dose of ALXN1210 or a subcutaneously administered placebo in 42 healthy subjects.

[0334] 1. Status of ongoing clinical trials Of the 161 screened subjects, 42 (26.09%) were randomly assigned to receive the investigational drug: placebo SC (n=6), ALXN1210 SC (n=24), and ALXN1210 IV (n=12) (Figure 48). None of the randomized subjects discontinued the trial early.

[0335] 2. Protocol deviations At least one protocol deviation was reported in 36 subjects (placebo SC: n=6; ALXN1210 SC: n=20; and ALXN1210 IV: n=10). Categories of protocol deviations included time frame deviations, subject compliance, failure to perform assessments, exclusion criteria, and dosage administration.

[0336] In two subjects of the ALXN1210 IV population, protocol deviations were assessed as significant. In one subject, evaluation of ADA, PK, PD, and clinical laboratory tests at day 29 was not performed because the subject did not attend follow-up appointments. In the other subject, PK and PD samples at day 71 were not collected because the subject did not attend follow-up appointments. These deviations were assessed as significant deviations due to the nature of the study design (PK-related primary endpoint), but they were not considered to affect the interpretation of results. None of the other protocol deviations were considered to affect the interpretation of results or the safety of the subjects. Serum pregnancy tests were negative in all subjects throughout the study.

[0337] 3. Pharmacokinetic evaluation, pharmacodynamic evaluation, and immunogenicity evaluation. All 42 randomized subjects received the investigational drug and were included in the safety set (Table 19). All of these subjects were also included in the PD set and the immunogenicity analysis set according to the definition. The 36 subjects in the safety set who received either ALXN1210 SC or ALXN1210 IV had sufficient serum concentration data and were able to calculate PK parameters, and were included in the PK set (Table 19).

[0338] Table 19: Analysis population (all participants were randomized) [Table 19]

[0339] 4. Demographic and other basic characteristics In the overall treatment population, the majority of subjects were male (66.7%) and Caucasian (69.0%), with a mean (±SD) age of 35.0 (±7.65) years. The mean (±SD) BMI for the entire population was 24.035 (±3.1582). Overall, the demographics were well-balanced across the treatment population (Table 20).

[0340] Table 20: Demographic Data - Descriptive Statistics by Treatment (Safety Set) [Table 20]

[0341] Previous drug use was reported in 5 individuals (20.8%) of the ALXN1210 SC population.

[0342] There were no reports of prior drug use in the placebo SC group or the ALXN1210 IV group.

[0343] The use of concomitant medications was reported in 3 (50.0%), 13 (54.2%), and 8 (66.7%) participants in the placebo SC, ALXN1210 SC, and ALXN1210 IV groups, respectively. The most frequently used concomitant medications were anilide for AE treatment, such as acetaminophen / paracetamol (15 participants), followed by multidrug combinations of progestogens and estrogens for contraception (7 participants). No concomitant medications were reported that were predicted to affect the results of this study.

[0344] No subjects received any non-pharmacological treatments or medical procedures. The total dose of ALXN1210 SC or placebo SC was administered by four intraventricular injections of 100 mg / 1 mL, respectively. The total dose of ALXN1210 IV was administered by intravenous infusion using an IV set with an in-line filter. All subjects received their assigned dose.

[0345] 5. List of pharmacokinetic, pharmacodynamic, and immunogenicity results and individual target data. PK analysis was performed on the PK set, which consisted of all subjects from a safety set who were administered either ALXN1210 SC or ALXN1210 IV and had sufficient serum concentration data to calculate PK parameters.

[0346] Figures 49–50 show the mean (±SD) serum concentration-time profiles of healthy subjects after ALXN1210 administration via seroconcentrate (SC) and intravenous (IV) administration (linear and log-linear scales). Plots of individual ALXN1210 serum concentrations and planned times are presented using a linear scale (Figure 49) and a log-linear scale (Figure 50), respectively.

[0347] The pharmacokinetic parameters of ALXN1210 after SC administration and IV administration are summarized in Table 21. A total of 24 subjects received ALXN1210 SC administration; t max The median (range) was 169.8 hours (96.0 to 508.1 hours). Geometric mean (CV%) t 1 / 2 The elimination times for ALXN1210 were similar for SC administration and IV administration, at 31.3 (13.6) days and 29.9 (15.4) days, respectively. ALXN1210 elimination was also similar between the IV and SC pathways (Figure 49).

[0348] Table 21: Summary of pharmacokinetic parameters of ALXN1210 (pharmacokinetic set) [Table 21]

[0349] Table 22 summarizes the absolute bioavailability of ALXN1210 SC. PK parameters (C) of ALXN1210 SC max AUC t , and AUC ∞ The results were compared to the baseline (ALXN1210 IV) by statistical analysis using a mixed model after logarithmic transformation of the data. max The GMR was 26.1% (95% CI: 21.3, 32.0). AUC ∞ Based on the estimated GMR (SC / IV), the absolute bioavailability of ALXN1210 SC was 60.4% (95% CI: 49.7, 73.3).

[0350] Table 22: Statistical analysis of absolute bioavailability of ALXN1210 subcutaneously (pharmacokinetic set) [Table 22]

[0351] PD analysis was performed on the PD set, which consisted of all subjects from a safety set that had sufficient free C5 concentration and total C5 concentration data, as well as cRBC hemolysis data.

[0352] Figure 51 shows the mean (±SD) percentage change in free C5 serum concentration over time from baseline for subjects treated with placebo SC, ALXN1210 SC, and ALXN1210 IV. After administration of placebo SC, mean free C5 remained relatively constant. A single dose of ALXN1210 IV 400 mg resulted in immediate and nearly complete free C5 inhibition (≥99%) from IV administration to day 8. A single dose of ALXN1210 SC 400 mg also resulted in a decrease in free C5, but it was not to the same extent or as immediate as that observed after IV administration. The maximum mean inhibition rate of free C5 (77%) after administration of ALXN1210 SC occurred one week after administration.

[0353] The duration and extent of the mean decrease in free C5 concentration were exposure-dependent. Figure 52 shows the mean (±SD) percentage change in total serum C5 concentration over time from baseline for subjects treated with placebo SC, ALXN1210 SC, and ALXN1210 IV. After placebo SC administration, mean total C5 remained relatively constant. However, administration of a single dose of ALXN1210 400 mg resulted in maximum mean increases of 82% and 107% in total C5 after SC and IV administration, respectively, compared to baseline.

[0354] Figure 53 shows the mean (±SD) percentage change in chicken erythrocyte (cRBC) hemolysis over time from baseline for subjects treated with placebo SC, ALXN1210 SC, and ALXN1210 IV. Mean cRBC hemolysis remained relatively constant after placebo SC administration. A single dose of 400 mg of ALXN1210 IV immediately inhibited mean cRBC hemolysis, with a maximum mean reduction of 88%. A single dose of SC 400 mg also resulted in a reduction in cRBC hemolysis, but it was not comparable to or immediate to that observed with IV administration. A maximum mean inhibition of 29% cRBC hemolysis occurred approximately one week after ALXN1210 SC administration. The duration and degree of cRBC inhibition were exposure-dependent.

[0355] Immunogenicity analysis was performed on an immunogenicity analysis set. This set consisted of all subjects from the safety set from which ADA samples were collected before and after administration. Anti-drug antibody tests were performed before and after administration on days 15, 29, 57, 90, 120, 150, and 200.

[0356] One subject (subject 0344-185) in the ALXN1210 SC treatment population tested positive for ADA in both the baseline (pre-administration) sample and all post-administration samples. In this subject, all post-administration antibody titers were lower than the pre-administration titers. The positive anti-drug antibody reaction in this subject was not considered clinically significant or related to ALXN1210. Therefore, this subject is not included in the immunogenicity summary presented below.

[0357] A total of four subjects (3 / 23 [13%] of the ALXN1210 SC population and 1 / 12 [8.3%] of the ALXN1210 IV population) developed ADA under administration of the study drug. In the ALXN1210 SC population: The first subject was ADA-positive on days 57, 90, 120, 150, and 200. All ADA-positive values ​​were positive for eculizumab cross-reactivity. The second subject was ADA-positive on days 29, 57, 90, 120, 150, and 200. All ADA-positive values ​​were positive for eculizumab cross-reactivity, except for a negative value on day 90. The third subject was ADA-positive on days 90, 120, 150, and 200. All ADA-positive values ​​were positive for eculizumab cross-reactivity.

[0358] In the ALXN1210 IV population: One subject was ADA-positive on days 15, 29, 90, 120, 150, and 200. All ADA-positive values ​​were negative for eculizumab cross-reactivity.

[0359] For both SC administration and IV administration, the earliest post-administration positive ADA reactions were observed on days 29 and 15, respectively. ADA titers in ADA-positive samples were low, ranging from <1.0 to 27. In most SC-administered ADA-positive samples, ADA cross-reactive with eculizumab. After IV administration, ADA did not cross-reactive with eculizumab. All ADA-positive subjects remained positive until the end of the follow-up period. A formal assessment of the effects of ADA on PK and PD could not be performed due to the small number of ADA-positive subjects. Examining the limited individual PK and PD outcomes in these subjects suggests that ALXN1210 does not have an apparent immunogenic effect on PK or PD.

[0360] 6. Conclusions on pharmacokinetics, pharmacodynamics, and immunogenicity Median (range) t max The elimination time after SC injection was 169.8 (96.0–508.1 hours). The geometric mean terminal phase elimination half-lives were similar after administration of ALXN1210 SC and IV, at 31.3 days and 29.9 days, respectively.

[0361] C max The estimated GMR (SC / IV) was 26.1% (95% CI: 21.3, 32.0). AUC ∞ The absolute bioavailability of ALXN1210 SC based on the estimated GMR (SC / IV) was 60.4% (95% CI: 49.7, 73.3).

[0362] The degree and duration of the PD response, as assessed by free C5 serum concentration, total C5 serum concentration, and cRBC hemolysis, were exposure-dependent. A single dose of ALXN1210 IV 400 mg resulted in immediate and nearly complete free C5 inhibition (≥99%) from administration to day 8. A single dose of ALXN1210 SC 400 mg administered as four 100 mg SC injections also resulted in a decrease in free C5, but it was not to the same extent or as immediate as that observed after IV administration. The maximum mean inhibition of free C5 was 77%, occurring approximately one week after SC administration. A 400 mg dose resulted in maximum mean increases of 82% and 107% in total C5 after SC administration and IV administration, respectively, compared to baseline. A single dose of ALXN1210 IV 400 mg resulted in immediate inhibition of mean cRBC hemolysis, with a maximum mean reduction of 87%. A single dose of ALXN1210 SC 400 mg also resulted in a reduction of cRBC hemolysis, but it was not comparable to or immediate compared to IV administration. The maximum mean inhibition of cRBC hemolysis in 29% occurred approximately 8 days after SC administration.

[0363] ADA (antibody-associated dysphatidylase) during administration of the study drug was reported in 3 out of 23 subjects (13%) and 1 out of 12 subjects (8.3%) in the ALXN1210 SC (single-course) and ALXN1210 IV (intravenous) populations, respectively, with low ADA titers ranging from <1.0 to 27. The earliest post-administration ADA response was observed on days 29 and 15, respectively, for both SC and IV administrations. After SC administration, ADA cross-reactive with eculizumab in most ADA-positive samples. After IV administration, ADA did not cross-reactive with eculizumab. All ADA-positive subjects remained positive until the end of the follow-up period. There was no apparent immunogenicity of ALXN1210 on PK or PD.

[0364] In the ALXN1210 SC treatment population, one additional subject tested positive for ADA in both the baseline (pre-administration) sample and all post-administration samples. In this subject, all post-administration antibody titers were lower than the pre-administration titers. The positive anti-drug antibody response in this subject was not associated with ALXN1210.

[0365] 7. Degree of exposure All subjects who received a single dose of the investigational drug were included in the safety set (N=42): placebo SC (n=6), ALXN1210 SC (n=24), and ALXN1210 IV (n=12). Each subject assigned to ALXN1210 IV received a total infusion volume (80 mL) of the investigational drug. In one subject, the infusion was interrupted for several minutes because the pump was not programmed with sufficient time to complete the infusion. Each subject who received either ALXN1210 SC or placebo SC received a total dose (4 mL) of the investigational drug.

[0366] 8. Adverse Events Across all three treatment groups, 35 out of 42 subjects (83.3%) experienced 75 TEAEs (all Grade 1). The proportion of subjects with at least one TEAE was 91.7%, 83.3%, and 79.2% in the ALXN1210 IV, placebo SC, and ALXN1210 SC groups, respectively. There were no reported deaths or SAEs during the study. No subjects discontinued the study or withdrew from the study due to TEAEs (Table 23). All TEAEs resolved during the course of the study. The majority of TEAEs required no treatment at all, and no subjects required non-pharmacological intervention at any point.

[0367] Table 22: Adverse Events (TEAEs) under Investigational Drug Administration - Overall Summary (Safety Set) [Table 23]

[0368] In total, 75 TEAEs were reported in 35 subjects. Across the entire treatment population, the most frequently reported TEAEs were nasopharyngitis (23 out of 42 subjects, 54.8%) and headache (7 out of 42 subjects, 16.7%). All TEAEs are summarized in Table 23 by systemic classification (SOC) and basic terminology for each treatment.

[0369] Table 23: Frequency table of adverse events under administration of the investigational drug - major classification by organ and basic terminology (safety set) [Table 24-1] [Table 24-2]

[0370] The majority of TEAEs (72 / 75 TEAEs, 96%) were considered unrelated to ALXN1210 treatment. Across the entire treatment population, 3 out of 42 subjects (7.1%) reported three TEAEs. These were related to ALXN1210 treatment ("probably related") and were assessed by the principal investigator as Grade 1 (mild): (1) an upper respiratory tract infection in one subject in the ALXN1210 SC population, (2) a migraine in one subject in the ALXN1210 SC population, and (3) a headache in one subject in the ALXN1210 IV population. All 75 TEAEs were classified as Grade 1 (mild). No subjects who experienced SAEs died, and no subjects discontinued the study or the investigational drug due to TEAEs.

[0371] In general, the mean values ​​for blood tests, coagulation tests, blood chemistry tests, urine tests, and urine chemistry tests were within the normal range, and there were no clear trends in the mean changes from the normal range.

[0372] The majority of participants in the study had normal values ​​(i.e., within the respective reference ranges) for parameters in blood tests, urine tests, coagulation tests, blood chemistry tests, and urine chemistry tests. No clear trends in deviations were observed across the treatment population. A shift from normal values ​​at baseline to abnormal values ​​(grade 1 [mild] or grade 2 [moderate]) was observed in some laboratory parameters during the study. However, these were not considered clinically significant. Most deviations were transient and resolved during the study.

[0373] During the study, a shift to Grade 3 outliers was reported in three subjects from the ALXN1210 SC population. No shifts to Grade 3 outliers were reported as AEs.

[0374] Initially, a decrease in neutrophil count was reported in one patient. In this patient, the baseline neutrophil count was 3.77 × 10⁹ / L. The neutrophil count evaluated on day 43 was 0.95 × 10⁹ / L (normal range: 2.0 to 7.5 × 10⁹ / L). The neutrophil count returned to the normal range on day 57.

[0375] Elevated potassium levels (normal range: 3.5–5.1 mmol / L) were reported in two subjects. One subject had a baseline potassium level of 4.5 mmol / L, and the potassium level assessed at day 150 was 6.1 mmol / L. Repeated potassium level measurements on the same day (unscheduled visit) were within the normal range. The other subject had a baseline potassium level of 4.6 mmol / L, and the potassium level assessed at day 90 was 6.4 mmol / L. This subject exhibited abnormal potassium levels at screening (ranging from 5.2–6.2 mmol / L between different screening visits) and throughout most of the trial visits. The increase in potassium levels was transient. Recorded values ​​were within the normal range at days 150 and 200.

[0376] For each individual subject, there were no observable changes from baseline in vital signs, and similarly, there were no clinically significant and consistently observed abnormalities in vital signs.

[0377] No subjects had clinically significant physical examination findings other than those reported as adverse events (AEs). There were no significant mean changes from baseline in ECG or telemetry monitoring results.

[0378] Changes in the QT interval were corrected using Fridericia's formula (QTcF). In one subject in the placebo SC population, a mean QT interval of >500 msec was observed at screening (510.0 msec), day 2 (508.7 msec), day 150 (516.6 msec), and day 200 (612.9 msec). In the same subject, the mean QTcF intervals were 449.7 msec, 443.7 msec, 451.9 msec, and 501.3 msec at screening and at days 2, 150, and 200, respectively. The increases in QT and QTcF intervals were not considered clinically significant in this female subject receiving placebo. These changes were also not reported as adverse events (AEs). No significant changes from baseline in mean QT and QTcF intervals were observed during the study.

[0379] Evaluation of the infusion or injection site was performed within 15 minutes ± 15 minutes of the SOI at 30 minutes, 2 hours, 4 hours, 8 hours, and on day 2 (48 hours) and day 3 (72 hours, a total of 6 evaluations). Induration or reaction less than 1 cm was not considered an adverse event unless it persisted for more than 24 hours. Erythema was observed 30 minutes after EOI in 5 out of 24 subjects in the ALXN1210 SC population. In one subject, minimal erythema (3 mm) was present at 2 hours after injection at 2 out of 4 injection sites, but not at the final time. Minimal induration or swelling (10 mm) was reported 30 minutes after EOI in 1 out of 24 subjects in the ALXN1210 SC population, but was not observed at the final evaluation. However, none of these met the criteria specified in the protocol for being considered an adverse event. Pain at the infusion or injection site was graded by subject using a Visual Analog Scale (VAS) (0-100 mm). For most infusions and injections, injection site pain was graded as 0 mm in all assessments. Two subjects in the SC group reported transient pain of 3-5 mm on day 1, and three subjects in the IV group reported minimal pain (1-5 mm) after infusion.

[0380] 9. Conclusions regarding safety All subjects who received a single dose of the investigational drug were included in the safety set (N=42) (placebo SC, ALXN1210 SC, and ALXN1210 IV). Across all three treatment groups, 35 / 42 subjects (83.3%) experienced 75 TEAEs. Only 3 / 75 TEAEs (4%) were considered ALXN1210-related, while 72 / 75 (96%) were considered unrelated to ALXN1210 treatment. All TEAEs were mild (grade 1) and resolved during the course of the study. The majority of TEAEs required no treatment at all, and no subjects required non-pharmacological intervention at any point. The most frequently reported TEAEs were nasopharyngitis (23 / 42 subjects, 54.8%) and headache (7 / 42 subjects, 16.7%).

[0381] There were no deaths or SAEs during the study. No TEAEs leading to discontinuation of the study drug or withdrawal of subjects from the study were reported. In general, there were no clinically significant changes in laboratory parameters, vital signs, physical examination, ECG, or telemetry during the study or follow-up period. There was no clinical evidence of hypersensitivity during or after a single dose of either SC injection or IV infusion. In subjects with ADA-positive results, there were no clinical signs or symptoms associated with allergic reactions or hypersensitivity.

[0382] 10. Discussion and Overall Conclusion The objective of this Phase I trial was to evaluate the safety, tolerability, pharmacokinetics, disease progression, and immunogenicity of ALXN1210 IV 400 mg or placebo SC injection as a single dose compared to a single dose of ALXN1210 SC 400 mg. A total of 42 subjects were randomized and received either placebo SC (n=6), ALXN1210 SC (n=24), or ALXN1210 IV (n=12).

[0383] ALXN1210 at a dose of 400 mg was well-tolerated in healthy subjects via the SC administration route. AUC ∞The absolute bioavailability of ALXN1210 SC based on GMR of the estimated (SC / IV) was 60.4% (95% CI: 49.7, 73.3). Geometric mean t 1 / 2 The estimated durations were 31.3 days and 29.9 days after ALXN1210 SC administration and IV administration, respectively. The degree and duration of the PD response, as assessed by free C5 serum concentration, total C5 serum concentration, and cRBC hemolysis, were exposure-dependent.

[0384] Antidrug antibodies were reported in 3 out of 23 subjects (13%) and 1 out of 12 subjects (8.3%) in the ALXN1210 SC population and ALXN1210 IV population, respectively, with ADA positive titers ranging from <1.0 to 27. The earliest post-administration reactions were observed on days 29 and 15, respectively, for both SC and IV administrations. In most ADA-positive samples after SC administration, ADA cross-reactive to eculizumab. There were no clinical signs or symptoms consistent with allergic reactions or hypersensitivity (including anaphylaxis) in subjects showing a positive ADA reaction. Furthermore, no apparent impact on ALXN1210 PK or PD was identified.

[0385] During this study, no unexpected safety concerns were observed in any treatment group. No deaths or SAEs occurred, and no subjects experienced any TEAEs leading to discontinuation of the study or withdrawal from the study.

[0386] Array Overview [Table 25-1] [Table 25-2] [Table 25-3]

Claims

[Claim 1] The method described in the specification.

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