Novel surfactants and their use

Abstract

The present invention provides aqueous pharmaceutical compositions comprising a compound of formula (I) as defined herein, as well as novel methods and uses of the compound as a surfactant in aqueous antibody compositions.

Description

[Technical Field]

[0001] Field of Invention The present invention relates to the field of aqueous pharmaceutical antibody formulations stabilized against the formation of visible particles, including, for example, antibody aggregates, antibody aggregates, and particles based on silicone oil or free fatty acids. [Background technology]

[0002] Background of the Invention Surfactants are important excipients in protein formulations because they protect unstable proteins from interfacial stress that can lead to protein aggregation. Proteins such as monoclonal antibodies (mAbs) are administered parenterally, which limits the selection of surfactants, including not only one of the most commonly used surfactants, polysorbate 20 (PS20), but also polysorbate 80, poloxamer 188, and Kolliphor / Solutol® HS 15 (polyoxyethylene ester of 12-hydroxystearic acid). PS20 can degrade during product storage by oxidative degradation or enzymatic hydrolysis. The latter, in particular, yields free fatty acids (FFAs) as degradation products, which can precipitate in solution and subsequently form microscopically visible and visible particles. Under conditions typical of biopharmaceutical formulations, FFAs can precipitate even below their temperature-dependent solubility limits, although the point of particle precipitation, even with well-characterized degradation profiles, is poorly understood.

[0003] Therefore, there is a need for alternative surfactants that do not have problems with inherent stability and adsorption behavior at pharmaceutically relevant interfaces. In particular, there remains a need to investigate novel / alternative surfactants to mitigate existing problems with established surfactants for parenteral administration in order to expand the toolbox for formulation development while ensuring optimal formulation stability. [Overview of the project]

[0004] This invention solves this problem by providing a compound for novel use as a surfactant in aqueous antibody preparations, preferably aqueous compositions of therapeutic monoclonal antibodies. [Brief explanation of the drawing]

[0005] (Figure 1) Thermal stereostructure stability of mAbs in the presence of surfactants (only mAb formulations and surfactants that have a significant influence on thermal stereostructure, as well as reference substances, are shown). The figure shows the starting temperature (T on (left) and melting temperature (T m1 The average of the two individual measurements (right) is shown. Surfactants with shaded values ​​show a significant decrease in thermal stability compared to control formulations without surfactants (w / o) and benchmark surfactants (PS20 and Px188). (Figure 2) Visible particles of the fpMab formulation detected in the EP box after shaking at 5°C for 7 days, as shown in the heatmap: Class (I): 0 particles, Class (II): 1 to 3 particles in up to 1 of 3 vials, Class (III): ≥4 particles in 1 or 1 to 2 particles in 2 of 3 vials, and Class (IV): ≥5 particles in ≥2 vials or ≥2 particles in ≥3 vials. (Figure 3) Visible particles detected in the EP box after different storage times declared in weeks (w) under different conditions: Class (I): 0 particles, Class (II): 1-4 particles in vials up to 30%, Class (III): >5 particles in vials <30%, or 1 particle in vials <50%, and Class (IV): >5 particles in vials >30%, or >2 particles in vials >40%. Classes are marked with different gray intensities (darker gray indicates a higher class). Formulations in prefillable syringes (pfs) are marked accordingly. (Figure 4) Turbidity of mAb formulations with different storage conditions (39 weeks at 5°C, 26 weeks at 25°C / 65% rH, and 13 weeks at 40°C / 75% rH, compared to initial values) and surfactant concentrations. Turbidity is measured in specific turbidity units (NTU). Only turbidity at the end of the storage time is shown in the heatmap. (Figure 5) Soluble aggregate levels given as HMW increase (area %) after different storage conditions and surfactant concentrations (0.06 (first dot), 0.2 (second dot), and 0.6 mg / mL (third dot, respectively): (A) 5°C; (B) 25°C / 60% rH, (C) 4 weeks (■) and 12 weeks at 40°C / 75% rH TIFF2026509585000001.tif4128, 26 weeks (△), and 39 weeks TIFF2026509585000002.tif4128 was compared to the initial value (-). (Figure 6) Cumulative number of microscopically visible particles ≥ 10 μm / mL for formulations containing surfactants at 0.06, 0.2, and 0.6 mg / mL. SVP count after storage at 5°C for 39 weeks, 25°C / 60% rH for 26 weeks, and 40°C / 75% rH for 12 weeks. Darker colors indicate a higher number of SVPs. (Figure 7) Characterization of selected formulation particles by FTIR, primarily to confirm the presence of proteins and protein-PDMS particles. Measurements were taken at the final stage of the stability test. (Figure 8) Selected FlowCam images of protein-PDMS particles (PPPs) after 13 weeks of storage at 40°C / 75% rH in a Mab2 formulation containing poloxamer. (Figure 9) Visible particles detected in the EP box after 1 week of shaking (sk) and 5 freeze-thaw cycles (F / T): Class (I): 0 particles, Class (II): 1-4 particles in vials up to 30%, Class (III): >5 particles in vials <30%, or 1 particle in vials <50%, and Class (IV): >5 particles in vials >30%, or >2 particles in vials >40%. (Figure 10) Turbidity of mAb formulations with different stress conditions (1 week of shaking at 5 / 25°C (sk) and 5 freeze-thaw cycles (F / T)) and surfactant concentrations. Turbidity is measured in specific turbidity units (NTU). Samples marked with * were not measured due to the very high particle volume. (Figure 11) Cumulative number of microscopically visible particles ≥ 10 μm / mL for formulations containing surfactants at 0.06, 0.2, and 0.6 mg / mL. SVP count after 7 days of shaking at 5°C, 25°C / 60% rH, and 5 freeze-thaw cycles at -20 / 5°C (F / T). *Samples not measured due to too many particles (the particle limit of the method was reached). (Figure 12) Soluble aggregate levels given as increase in HMW (area %) after different stress conditions (7 days of shaking at 5°C, 7 days of shaking at 25°C, and 5 freeze / thaw -20 / 5°C cycles (F / T)) and surfactant concentrations (0.06, 0.2, and 0.6 mg / mL): bsMab2, IgG4, and MP compared to initial values ​​(1st row). [Modes for carrying out the invention]

[0006] Detailed description of the invention In one embodiment, the present invention relates to an aqueous pharmaceutical composition comprising an antibody and a surfactant, wherein the surfactant is of formula (I) H-(-OE) n -(OBu) m -X-(BuO) m -(EO) n -H (I) (In the formula, -OE is ethoxy, -OBu is n-butoxy, -X- is -On-butylene-O- or The filename is TIFF2026509585000003.tif28128. n is 40, 48, or 68. (m is 11, 12, or 16) The present invention provides a composition which is a compound of [the compound].

[0007] In another embodiment, the present invention provides a composition as defined above, -X- is -O-n-butylene-O-, n is 40 or 48, m is 12 or 16.

[0008] In another embodiment, the present invention provides a composition as defined above, -X- is -O-n-butylene-O-, n is 40, m is 16.

[0009] In another embodiment, the present invention provides a composition as defined above, -X- is -O-n-butylene-O-, n is 48, m is 12.

[0010] In another embodiment, the present invention provides a composition as defined above, -X- is TIFF2026509585000004.tif28128, n is 68, m is 11.

[0011] For example, compounds of formula (I) containing reference compounds used herein with different values of "m" and "n" may also be designated as "butron(s)". In one embodiment, the compound of formula (I) has the more specific formula (I-a) TIFF2026509585000005.tif31129, wherein "m" and "n" have the meanings given for formula (I). In one embodiment, in formula (I-a), m is 12 and n is 48.

[0012] The compounds of formulas (I) and (Ia) are polymers and can generally be obtained by methods known to those skilled in the art. Those skilled in the art will recognize that the values ​​given above for “m” and “n” are part of a range for the chemical synthesis involved in the production of the compounds of formula (I). In one embodiment, the specific values ​​given above for “m” and “n” represent general values ​​within that range. In another embodiment, the specific values ​​given herein for “m” and “n” may deviate by up to 4 and up to 6, respectively. For example, in formula (I) or (Ia), if -X- is -On-butylene-O-, then n is 48±6, or ±5, or ±4, or ±3, or ±2, or ±1, and m is 12±4, or ±3, or ±2, or ±1. In yet another embodiment, the weight % (%(w / w)) of the ethoxy moiety (-OE) in the compound of formula (I) or (Ia) is 60-80%(w / w), or 60-75%(w / w), or 60-70%(w / w).

[0013] Furthermore, as described above, coinciding with the ranges of "m" and "n", those skilled in the art will know that the synthesis of a compound of formula (I) or (Ia) may result in a mixture of products within a range of molecular weights (MW). In one embodiment, the specific molecular weights shown herein, for example in Table 1, are those of the dominant product in the mixture. In another embodiment, the compound of formula (I) or (Ia) has a molecular weight in the range of 4000 to 10000 g / mol, or 4000 to 7000 g / mol, or 5000 to 7000 g / mol, or 5500 to 6500 g / mol, or 5900 to 6100 g / mol. In yet another embodiment, the compound of formula (I) or (Ia) has a molecular weight in the range of 5900 to 6100 g / mol and a weight percentage of ethoxy units in the range of 60 to 70% (w / w).

[0014] The term "aqueous pharmaceutical composition" means an aqueous composition, formulation, or dosage form for pharmaceutical use. In one embodiment, the liquid pharmaceutical composition is for parenteral administration of a therapeutic antibody. In another embodiment, the liquid pharmaceutical composition according to the present invention comprises one or more therapeutic antibodies together with pharmaceutically acceptable excipients or carriers. Such excipients are generally known to those skilled in the art.

[0015] The term “excipient” has the usual meaning known to those skilled in the art with respect to parenteral antibody compositions. In one embodiment, the term “excipient” refers to components in a pharmaceutical composition or formulation other than the active ingredient that are nontoxic to the subject. Examples of excipients include, but are not limited to, buffers, stabilizers including antioxidants, or preservatives.

[0016] The term "pharmaceutical composition" refers to a preparation, formulation, or dosage form that is in a form that enables the biological activity of the active ingredient contained herein to be effective, and that does not contain additional ingredients that are unacceptably toxic to the subject to which the pharmaceutical composition is administered.

[0017] The term "pharmaceutically acceptable carrier" refers to a component in a pharmaceutical composition or formulation other than the active ingredient that is non-toxic to the subject. pharmaceutically acceptable carriers include, but are not limited to, excipients as defined herein.

[0018] The term "buffering agent" is well known to those skilled in the art of organic chemistry or pharmacy, for example, in the development of pharmaceutical preparations. As used herein, buffering agents refer to acetate, succinate, citrate, arginine, histidine, phosphate, tris, glycine, aspartate, and glutamate buffer systems. The pH range provided by the buffering agent is 4 to 8, preferably 4.5 to 7.5, and more preferably 5 to 7. Furthermore, in this embodiment, the histidine concentration of the buffering agent is 5 to 50 mM, preferably 10 to 25 mM.

[0019] The term “stabilizer” is well known to those skilled in the art of organic chemistry or pharmaceuticals, for example, in the development of pharmaceutical preparations. The stabilizers according to the present invention are selected from the group consisting of sugars, sugar alcohols, sugar derivatives, or amino acids. In one embodiment, the stabilizer is selected from one or more of the following groups: (1) sucrose, trehalose, cyclodextrin, sorbitol, mannitol, glycine, and / or (2) methionine, and / or (3) arginine, or lysine. In one embodiment, the stabilizer may be used at concentrations of up to 500 mM, or up to 350 mM, or up to 250 mM, or up to 150 mM (including 150 mM). In yet another embodiment, the concentration of the stabilizer relates to group (1) up to 500 mM, or up to 350 mM, or up to 250 mM, or up to 150 mM (including 150 mM), group (2) 5 to 40 mM, or 5 to 30 mM, or 5 to 25 mM, or / and group (3) up to 350 mM, or up to 250 mM.

[0020] In this specification, the term “antibody” is used in its broadest sense and encompasses a diverse class or structure of antibodies, including but not limited to monoclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), antibody-cytokine fusion proteins (fpMab), and antibody fragments, as long as they exhibit the desired antigen-binding activity. In one embodiment, the fusion protein in the antibody-cytokine fusion protein is IL-2. In one embodiment, the term “antibody,” as used herein, refers to a multimeric protein, preferably a pentameric protein.

[0021] In one embodiment of the present invention, the antibody is a monoclonal antibody. The term “monoclonal antibody” is known to those skilled in the art. In one embodiment, the term “monoclonal antibody” refers to an antibody derived from a single clone, including any eukaryote, prokaryote, or phage clone, and not from the method by which it is produced.

[0022] In one embodiment, the antibodies are alemtuzumab (LEMTRADA®), atezolizumab (TECENTRIQ®), bevacizumab (AVASTIN®), cetuximab (ERBITUX®), panitumumab (VECTIBIX®), pertuzumab (PERJETA®, 2C4, Omnitarg), trastuzumab (HERCEPTIN®), tositumomab (Bexxar®), absiximab (REOPRO®), and adalimumab (HUMIRA®). Apolizumab, aselizumab, atlizumab, bapineozumab, basiliximab (SIMULECT®), babituximab, belimumab (BENLYSTA®), briankinumab, canakinumab (ILARIS®), sedelizumab, certolizumab pegol (CIMZIA®), sidofcituzumab, cizutuzumab, sixtumumab, crazakizumab, crenezumab, daclizumab (ZENAPAX®), dalotuzumab, denosumab (PROLIA®, XGEVA®), eclizma Zumab (SOLIRIS®), efalizumab, epratuzumab, erulizumab, emicizumab (HEMLIBRA®), felbizumab, fontrizumab, gantenerumab, golimumab (SIMPONI®), ipilimumab, imugatuzumab, infliximab (REMICADE®), rabetuzumab, lebrikizumab, lexatumumab, lintuzumab, lucatumumab, rulizumab pecol, lumuretuzumab, mapatumumab, matsuzumab, mepolizumab, mogamulizumab, motabizumab, motobizumab, mu Lonomab, natalizumab (TYSABRI®), necitumumab (PORTRAZZA®), nimotuzumab (THERACIM®), norovizumab, numavizumab, obinutuzumab (GAZYVA®), olokizumab, omalizumab (XOLAIR®), onarutuzumab (also known as MetMAb), palivizumab (SYNAGIS®), pascolizumab, pecufcituzumab, pectuzumab, pembrolizumab (KEYTRUDA®), pexerizumab, priliximab,Larivizumab, ranivizumab (LUCENTIS®), reslibizumab, reslizumab, recibizumab, lobatumumab, lontarizumab, loberizumab, luprizumab, sarilumab, secukinumab, cerivantuzumab, cifalimumab, cibrotuzumab, siltuximab (SYLVANT®), ciprizumab, sontuzumab, The "antibody product" is selected from tadocizumab, talizumab, tefivazumab, tocilizumab (ACTEMRA®), tralizumab, tuccituzumab, umabizumab, urtoxazumab, ustekinumab (STELARA®), vedolizumab (ENTYVIO®), bicilizumab, zanorimumab, and saltumumab.

[0023] An "antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody that binds to an antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv and scFab); single-domain antibodies (dAb); and multispecific antibodies formed from antibody fragments. For an overview of specific antibody fragments, see Holliger and Hudson, Nature Biotechnology 23:1126-1136 (2005).

[0024] The "class" of an antibody refers to the type of constant domain or constant region held by its heavy chain. Antibodies exist in five main classes: IgA, IgD, IgE, IgG, and IgM, some of which can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. In certain embodiments, the antibody is of the IgG1 isotype. In certain embodiments, the antibody is of the IgG1 isotype with P329G, L234A, and L235A mutations to reduce the effector function of the Fc region. In other embodiments, the antibody is of the IgG2 isotype. In certain embodiments, the antibody is of the IgG4 isotype with the S228P mutation in the hinge region to improve the stability of the IgG4 antibody. The heavy chain constant domains corresponding to different classes of immunoglobulins are called a, d, e, g, and m, respectively. The light chain of an antibody can be assigned to one of two types, called kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain. In one embodiment, the antibody according to the present invention is an IgG1 and / or IgG4 antibody.

[0025] In one embodiment, any of the antibodies according to the present invention is a human antibody or a humanized antibody. A "human antibody" is an antibody produced by a human or human cell, or an antibody having an amino acid sequence corresponding to the amino acid sequence of a non-human antibody that utilizes a sequence encoding a human antibody, such as the human antibody repertoire. The human antibody of the present invention may contain amino acid residues not encoded by a human germline immunoglobulin sequence (for example, mutations introduced by random or site-directed mutagenesis in vitro, or by somatic mutation in vivo).

[0026] A "humanized" antibody refers to a chimeric antibody containing amino acid residues derived from non-human CDRs and amino acid residues derived from human FRs. In certain embodiments, a humanized antibody comprises substantially all of at least one, typically two, variable domains, with all or substantially all of the CDRs corresponding to the CDRs of a non-human antibody and all or substantially all of the FRs corresponding to the FRs of a human antibody. A humanized antibody may optionally also include at least a portion of an antibody constant region derived from a human antibody. The "humanized form" of an antibody, e.g., a non-human antibody, refers to the antibody that has been humanized.

[0027] In one embodiment, the present invention provides a composition as defined herein, wherein the antibody is present at a concentration that provides its desired pharmaceutical activity and an acceptable safety profile. In another embodiment, the present invention provides a composition as defined herein, wherein the antibody is present at a concentration in the range of 1 to 220 mg / ml, preferably 5 to 180 mg / ml, or 5 to 100, or 5 to 25 mg / ml.

[0028] In another embodiment, the present invention provides a composition as defined herein, wherein the surfactant is present at a concentration of 0.001 to 1. mg / ml, or 0.01 to 1.0 mg / ml, or 0.06 to 1.0 mg / ml, or 0.06 to 0.6 mg / ml.

[0029] In another embodiment, the present invention provides a composition as defined herein, further comprising an additional pharmaceutically acceptable excipient.

[0030] In another embodiment, the present invention provides a compound of formula (I) for use as a surfactant in an aqueous antibody composition H-(-OE) n -(OBu) m -X-(BuO) m -(EO) n -H (I) (wherein -OE is ethoxy, -OBu is n-butoxy, -X- is -O-n-butylene-O- or The filename is TIFF2026509585000006.tif28128. n is 40, 48, or 68. (m is 11, 12, or 16) The compound is provided.

[0031] In yet another embodiment, the present invention provides a compound of formula (I) for use as a surfactant in an aqueous antibody composition, as defined above, wherein the compound stabilizes the antibody against aggregation. In one embodiment, the aggregation is the aggregation of several antibodies. In another embodiment, the aggregation is the aggregation of an antibody and PDMS.

[0032] In another embodiment, the present invention provides a compound of formula (I) for use as defined above, the compound preventing the formation of visible particles in an aqueous antibody composition.

[0033] In another embodiment, the present invention provides a compound of formula (I) for any use previously defined herein, -X- is -On-butylene-O-, n is either 40 or 48. m is either 12 or 16.

[0034] In another embodiment, the present invention provides a compound of formula (I) for any use previously defined herein, -X- is -On-butylene-O-, n is 40, m is 16.

[0035] In another embodiment, the present invention provides a compound of formula (I) for any use previously defined herein, -X- is -On-butylene-O-, n is 48, m is 12.

[0036] In another embodiment, the present invention provides a compound of formula (I) for any use previously defined herein, -X- is The filename is TIFF2026509585000007.tif28128. n is 68, m is 11.

[0037] In another embodiment, the present invention provides a compound of formula (I) for any use as previously defined herein, wherein the antibody is a monoclonal antibody.

[0038] In another embodiment, the present invention provides a compound of formula (I) for any use as previously defined herein, wherein the monoclonal antibody is of the IgG1 or IgG4 subclass.

[0039] In another embodiment, the present invention provides a compound of formula (I) for any use previously defined herein, wherein the surfactant is present at a concentration of 0.001 to 1.0 mg / ml, or 0.01 to 1.0 mg / ml, or 0.06 to 1.0 mg / ml, or 0.06 to 0.6 mg / ml.

[0040] In another embodiment, the present invention provides a compound of formula (I) for any use as previously defined herein, further comprising additional pharmaceutically acceptable excipients.

[0041] In another embodiment, the present invention provides a method for preventing the formation of visible particles in an aqueous antibody composition, wherein the method is based on formula (I) H-(-OE) n -(OBu) m -X-(BuO) m -(EO) n -H (I) (In the formula, -OE is ethoxy, -OBu is n-butoxy, -X- is -On-butylene-O- or The filename is TIFF2026509585000008.tif28128. n is 40, 48, or 68. (m is 11, 12, or 16) This includes the use of the compound.

[0042] In yet another embodiment, the present invention provides a method for preventing the formation of visible particles in an aqueous antibody composition, the method comprising the use of a compound of formula (I) as previously defined herein, -X- is -On-butylene-O-, n is either 40 or 48. m is either 12 or 16.

[0043] In yet another embodiment, the present invention provides a method for preventing the formation of visible particles in an aqueous antibody composition, the method comprising the use of a compound of formula (I) as previously defined herein, -X- is -On-butylene-O-, n is 40, m is 16.

[0044] In yet another embodiment, the present invention provides a method for preventing the formation of visible particles in an aqueous antibody composition, the method comprising the use of a compound of formula (I) as previously defined herein, -X- is -On-butylene-O-, n is 48, m is 12.

[0045] In yet another embodiment, the present invention provides a method for preventing the formation of visible particles in an aqueous antibody composition, the method comprising the use of a compound of formula (I) as previously defined herein, -X- is The filename is TIFF2026509585000009.tif28128. n is 68, m is 11.

[0046] In yet another embodiment, the present invention provides a method for preventing the formation of visible particles in an aqueous antibody composition, the method comprising the use of a compound of formula (I) as previously defined herein, wherein the antibody is a monoclonal antibody.

[0047] In yet another embodiment, the present invention provides a method for preventing the formation of visible particles in an aqueous antibody composition, the method comprising the use of a compound of formula (I) as previously defined herein, wherein the monoclonal antibody is of the IgG1 or IgG4 subclass.

[0048] In yet another embodiment, the present invention provides a method for preventing the formation of visible particles in an aqueous antibody composition, the method comprising the use of a compound of formula (I) as previously defined herein, the compound of formula (I) present at a concentration of 0.001 to 1.0 mg / ml, or 0.01 to 1.0 mg / ml, or 0.06 to 1.0 mg / ml, or 0.06 to 0.6 mg / ml.

[0049] In yet another embodiment, the present invention provides a method for preventing the formation of visible particles in an aqueous antibody composition, the method comprising the use of a compound of formula (I) as previously defined herein, before further comprising additional pharmaceutically acceptable excipients.

[0050] The uses and methods of the present invention are suitable for preventing the formation of visible particles in aqueous antibody compositions. In one embodiment, the formation of visible particles occurs during storage of the aqueous antibody composition. As used herein, the term “storage” means keeping the aqueous pharmaceutical product under conditions known to those skilled in the art, or as indicated, for example, in the package insert of a comparable commercial drug or in the corresponding summary of product characteristics. In one embodiment, the storage includes a period of time up to 6 months, or 12 months, or 18 months, or 24 months, or 30 months. In another embodiment, the storage includes maintaining the liquid pharmaceutical composition under conditions (e.g., temperature) also approved by the regulatory authority, up to its shelf life approved by the regulatory authority. In one embodiment, such shelf life and storage conditions may be found, for example, in the package insert or corresponding summary of product characteristics accompanying an approved protein-based drug. In another embodiment, any storage temperature within the approved shelf life is below 30°C. In yet another embodiment, the storage temperature is between 2 and 30°C. In yet another embodiment, the storage temperature is between 2 and 8°C.

[0051] The following is a set of clauses defining the present invention and its preferred embodiments and features: 1. An aqueous pharmaceutical composition comprising an antibody and a surfactant, wherein the surfactant is of formula (I) H-(-OE) n -(OBu) m -X-(BuO) m -(EO) n -H (I) (In the formula, -OE is ethoxy, -OBu is n-butoxy, -X- is -On-butylene-O- or The filename is TIFF2026509585000010.tif28128. n is 40, 48, or 68, and (m is 11, 12, or 16) A composition that is a compound of [the compound]. 2. -X- is -On-butylene-O-, n is 40 or 48, m is 12 or 16, and The composition described in Clause 1. 3. -X- is The filename is TIFF2026509585000011.tif28128. n is 68, and m is 11. The composition described in Clause 1. 4. The composition according to any one of clauses 1 to 3, wherein the antibody is a monoclonal antibody. 5. The composition according to Clause 4, wherein the monoclonal antibody is of the IgG1 or IgG4 subclass. 6. The composition according to any one of clauses 1 to 5, wherein the surfactant is present at a concentration of 0.001 to 1.0 mg / ml, or 0.01 to 1.0 mg / ml, or 0.06 to 1.0 mg / ml, or 0.06 to 0.6 mg / ml. 7. The composition according to any one of clauses 1 to 6, further comprising additional pharmaceutically acceptable excipients. 8. Formula (I) for use as a surfactant in aqueous antibody compositions H-(-OE) n -(OBu) m -X-(BuO) m -(EO) n -H (I) (In the formula, -OE is ethoxy, -OBu is n-butoxy, -X- is -On-butylene-O- or The filename is TIFF2026509585000012.tif28128. n is 40, 48, or 68, and (m is 11, 12, or 16) A compound of [this]. 9. A compound of formula (I) for use as described in Clause 8, wherein the compound stabilizes the antibody against aggregation. 10. A compound of formula (I) for use as described in Clause 9, wherein the aggregation is either the aggregation of several antibodies or the aggregation of an antibody and PDMS. 11. The compound of formula (I) for use as described in Clause 8, wherein the compound prevents the formation of visible particles in an aqueous antibody composition. 12. -X- is -On-butylene-O-, n is 40 or 48, and m is 12 or 16. A compound of formula (I) as described in any one of clauses 8 to 11. 13.-X- is The filename is TIFF2026509585000013.tif28128. n is 68, and m is 11. A compound of formula (I) as described in any one of clauses 8 to 11. 14. A compound of formula (I) for use as described in any one of clauses 8 to 13, wherein the antibody is a monoclonal antibody. 15. A compound of formula (I) for use as described in Clause 14, wherein the monoclonal antibody is of the IgG1 or IgG4 subclass. 16. A compound of formula (I) for use as described in either clause 8 or 15, wherein the surfactant is present at a concentration of 0.001 to 1.0 mg / ml, or 0.01 to 1.0 mg / ml, or 0.06 to 1.0 mg / ml, or 0.06 to 0.6 mg / ml. 17. Compounds of formula (I) for use as described in any one of clauses 8 to 16, further comprising additional pharmaceutically acceptable excipients. 18. A method for preventing the formation of visible particles in an aqueous antibody composition, wherein formula (I) H-(-OE) n -(OBu) m -X-(BuO) m -(EO) n -H (I) (In the formula, -OE is ethoxy, -OBu is n-butoxy, -X- is -On-butylene-O- or The filename is TIFF2026509585000014.tif28128. n is 40, 48, or 68, and (m is 11, 12, or 16) A method comprising the use of the compound. 19. -X- is -On-butylene-O-, n is 40 or 48, and m is 12 or 16. The method described in Article 18. 20.-X- is The filename is TIFF2026509585000015.tif28128. n is 68, and m is 11. The method described in Article 18. 21. The method according to any one of the clauses 18-20, wherein the antibody is a monoclonal antibody. 22. The method according to Clause 21, wherein the monoclonal antibody is of the IgG1 or IgG4 subclass. 23. The method according to any one of the claims 18 to 22, wherein the compound of formula (I) is present at a concentration of 0.001 to 1.0 mg / ml, or 0.01 to 1.0 mg / ml, or 0.06 to 1.0 mg / ml, or 0.06 to 0.6 mg / ml. 24. The method according to any one of the provisions of 18 to 23, further comprising additional pharmaceutically acceptable excipients. 25. The composition, use, and method described in any one of clauses 1 to 24, wherein the compound of formula (I) has a more specific formula (Ia) as defined herein.

[0052] The present invention will be further illustrated by the following non-limiting examples of work. [Examples]

[0053] material and method material The model antibody (mAb) containing the pentameric protein used in this study was provided by F. Hoffmann-La Roche (Basel, Switzerland): Glycan-modified IgG1 mab (gMab) formulated in 20 mM His-HCl buffer (Ajinomoto, Tokyo, Japan) containing 240 mM trehalose (Pfanstiehl Inc., Illinois, USA) at pH 6.0. IgG1 mab 1 (Mab1) formulated in 10 mM His-HCl buffer (Ajinomoto, Tokyo, Japan) containing 240 mM sucrose (Pfanstiehl Inc., Illinois, USA) at pH 6.0. IgG1 mAb 2 (Mab2) formulated in 20 mM His-HCl buffer (Ajinomoto, Tokyo, Japan) containing 200 mM trehalose (Pfanstiehl Inc., Illinois, USA) at pH 5.5. A bispecific mAb 1 (bsMab1) formulated in 20 mM His-HCl buffer (Ajinomoto, Tokyo, Japan) containing 240 mM sucrose (Pfanstiehl Inc., Illinois, USA) at pH 6.0. A bispecific mAb 2 (bsMab2) formulated in 10 mM His-HCl buffer (Ajinomoto, Tokyo, Japan) containing 240 mM sucrose (Pfanstiehl Inc., Illinois, USA) at pH 5.8. IgG4 mAb 2 (IgG4-2) formulated in 20 mM histidine buffer (Ajinomoto, Tokyo, Japan) containing 200 mM arginine succinate (Ajinomoto, Tokyo, Japan) at pH 5.7. A multimeric protein (MP, here a pentamer) formulated in 10 mM sodium phosphate buffer (Merck KGAA, Darmstadt, Germany; chem.Fabrik Budenheim, Budenheim, Germany) containing 5% (m / v) sorbitol (Merck KGAA, Darmstadt, Germany) at pH 7.5. Antibody-cytokine fusion protein mAb (fpMab) formulated in 20 mM His-HCl buffer (Ajinomoto, Tokyo, Japan) containing 240 mM sucrose (Pfanstiehl Inc., Illinois, USA) at pH 5.5.

[0054] Antibodies were tested in buffered aqueous solutions at concentrations ranging from 5 to 180 mg / mL.

[0055] The screened surfactants were provided by BASF (Ludwigshafen, Germany): Pluronic PE 10400 (Px334), Pluronic PE 10500 (Px335), butronic (Bux016, Bux017), butronic 4060 (Bux164), butronic 6060 (Bux190), butronic 6070 (Bux199), isosorbide alkoxylates (IA80, IA90), poly(methyl-butyl-methyl)oxazolines (Pz110, Pz120), and polyvinyl alcohol / polypropylene glycol (PVA / PPG). Polysorbate 20 (PS20; Croda International, Snais, UK) and Poloxamer 188 (Px188; BASF, Ludwigshafen, Germany) were used as guide reference substances. Table 1 provides a summary of the test compounds, including the reference surfactant.

[0056] [Table 1] *PO / OH=n-propoxy

[0057] All other reagents, including methanol (MeOH), anhydrous potassium dihydrogen phosphate (KH2PO4), anhydrous dipotassium hydrogen phosphate (K2HPO4), and potassium chloride (KCl), were analytical grade and obtained from Merck KGa, Darmstadt, Germany.

[0058] method Evaluation of the thermal structural stability of proteins in the presence of surfactants The stability of three-dimensional proteins was investigated using the Prometheus NT.Plex (NanoTemper Technologies GmbH, Munich, Germany). This instrument allows for label-free detection of intrinsic protein fluorescence changes from aromatic tryptophan and tyrosine residues using very small amounts of solution. Heat-induced protein unfolding was monitored by detecting emission shifts at 330 nm and 350 nm with optimized laser power of 7–12%. NanoDSF standard grade capillary tips (NanoTemper Technologies, Munich, Germany) were filled with 10 μL of freshly prepared formulations containing 25 mg / mL mAb compounded with one of the following specific surfactants: 0.01, 0.1, 1, or 10 mg / mL. Analysis was performed using five different mAbs (bsMab1, Mab1, gMab, bsMab2, and fpMab). Samples were heated from 20–95°C on a constant heating lamp at 0.5°C / min. PR.StabilityAnalysis software (NanoTemper Technologies, Munich, Germany) analyzes the melting curve at the start of (T on ) and the first transition point (T m1 The value was calculated automatically. The reported data is the average of two individual measurements.

[0059] Evaluation of protein stability after mechanical stress and thermal stability in the presence of surfactants. Surfactant performance screening was performed using surfactants at concentrations of 0.06, 0.2, and 0.6 mg / mL (for pre-screening with fpMab: 0.001, 0.01, 0.1, and 1 mg / mL) in formulations of different model mAbs as described in the materials section. All formulations were prepared, and the liquid samples were sterile filtered through a 0.22 μm Millex Sterivex® GV (Millipore, Bedford, USA) filter unit. Formulations containing fpMab, gMab, bsMab1, Mab1, bsMab2, IgG4, and MP were filled into 6 mL Type 1 glass vials and sealed with Φ20 mm Teflon® coated serum stoppers (Daikyo Seikou Co., Ltd., Tokyo, Japan). The sealed vials were crimped using aluminum caps with PP plates (Datwyler Holding AG, Altdorf, Switzerland). Furthermore, Mab1 was filled into 2.25 mL prefillable syringes BD Neopak® (pfs), and Mab2 was filled into 1 mL pfs (both from BD Medical-Pharmaceutical Systems, Franklin Lakes, USA), and each was sealed with a Teflon®-coated serum stopper for 2.25 mL or 1 mL of pfs (Daikyo Seikou Co., Ltd., Tokyo, Japan).

[0060] To evaluate the effect of surfactants on mAb stability, different interfacial stress conditions, including stirring and multiple freeze-thaw cycles, were applied to fpMab, bsMab2, IgG4, and MP formulations. Shaking was performed for 7 days at a constant 200 revolutions per minute (rpm) at 5°C and 25°C, with vials placed horizontally in a shaker (HS 260 Control Model; IKA Werke GmbH&Co.KG; Stauffen, Germany) and protected from light. Freeze-thaw (F / T) stress was performed by exposing vials to five consecutive cycles of freezing at -20°C and thawing at 5°C under control in an artificial climate chamber VTM 4004 (Votsch, Borken, D). For MP, only F / T stress was tested, and for fpMab, only shaking was performed for 7 days at 5°C.

[0061] Thermal stability data was prepared by storing liquid gMab, bsMab1, Mab1, and Mab2 formulations at 5°C for 9 months (mo), at 25°C / 60% relative humidity (rH) for 6 months, and at 40°C / 75% rH for 12 weeks. Samples were analyzed at the initial time (t0) and after 1 mo, 3 mo, 6 mo, and 9 mo of storage using the analytical methods described below.

[0062] visible particles (VP) Visual inspection was performed as described above, using a black and with box (Color Viewing Light 3 BASIC, JUST Normlicht, Weilheim, D) in accordance with Ph.Eur.2.9.20 [1]. The vials and pfs designated for visual inspection were analyzed after equilibration with RT. The vials and pfs were stored again after inspection, and the same vials were used for visual inspection at each time point. The number of particles was classified into the following four classes: Class (I) corresponds to 0 particles, Class (II) corresponds to 1 to 4 particles in up to 30% of vials, Class (III) corresponds to >5 particles in <30% of vials / 1 particle in <50% of vials, and Class (IV) corresponds to >5 particles in >30% of vials / >2 particles in >40% of vials.

[0063] Turbidity (milky and transparent) Turbidity was determined using a TL 2350 EPA turbidimeter (Hach Lange GmbH, Düsseldorf, Germany) calibrated with a StablCal® calibration kit (Hach Lange GmbH), as previously described in the literature and in accordance with Ph.Eur.2.2.1. The results are presented in turbidimetric units (NTU). [2, 3]

[0064] light shielding method Microscopic visible particles (SVP) were counted using the light-shielding method with a HIAC 9703+ liquid particle counting system (Beckman Coulter, Pasadena, USA) and PharmSpec 3 (Hach Lange GmbH) software. The measurement techniques applied were those specified in Ph.Eur.2.9.19[4] and USP <787> The method was adapted from the one described in [5]. After rinsing the system with the sample solution, it was run four times with a sample volume of 0.2 mL. The final cumulative particle count was obtained by calculating the mean ± SD (standard deviation) from the last three measurements. SVPs of 2, 5, 10, 25 and 50 μm or larger were detected and shown as the cumulative number per 1 mL of solution.

[0065] Background membrane imaging The number of subatomic particles (SVPs) in liquid samples was also measured using background membrane imaging (BMI) techniques with the Horizon system and Halo Lab software (both from Halo Lab, Burlingame, USA). A polycarbonate membrane plate with a pore size of 0.4 μm (Halo Lab, Burlingame, USA) was first washed with particle-free water. 40 μL of sample was packed into the washed filter (under a vacuum of 200 mbar for 2 minutes) and measured. The final cumulative particle count was obtained by calculating the mean ± SD from three measurements. SVPs of 2, 5, 10, 25, and 50 μm or larger were detected and expressed as the cumulative number per 1 mL of solution.

[0066] Size exclusion (ultra)fast chromatography (SE-(U)HPLC) Below, soluble mAb aggregates, monomers, and low molecular weight species (LMW), referred to as high molecular weight species (HMW), were analyzed by SE-HPLC for gMab, bsMab1, Mab1, and Mab2, or by SE-UHPLC for bsMab2, IgG4, and MP.

[0067] SE-HPLC: The system used consisted of an Alliance 2695 HPLC instrument equipped with a 2489 UV detector (both Waters Corporation, Milford, MA). The autosampler temperature was set to 5°C, and a total of 150 μg of mAbs were loaded onto the column. Separation was performed using a TSK G3000 SWXL, 7.8 × 300 mm column (Tosoh Bioscience, Stuttgart, Germany) at a constant oven temperature of 25°C. 200 mM K2HPO4 / KH2PO4 and 250 mM KCl pH 7.0 were used as the mobile phase at a flow rate of 0.5 mL / min. Signal detection was performed at a wavelength of 280 nm, and peak area percentages were calculated using Empower 3 Chromatography Data System software (Waters Corporation, Milford, MA).

[0068] SE-UHPLC: The system used consisted of a Thermo UltiMate 3000 UHPLC instrument equipped with a 3000 UV / vis detector (both from Thermo Fisher Scientific, Waltham, USA). The autosampler temperature was set to 10°C, and a total of 50 μg of mAbs were loaded into the system. Separation was performed using a TSK UP-SW3000, 4.6 × 300 mm column (Tosoh Bioscience, Stuttgart, Germany) at a constant oven temperature of 25°C, and a mobile phase of 200 mM K2HPO4 / KH2PO4 and 250 mM KCl pH 6.2 at a flow rate of 0.3 mL / min. Signal detection was performed at a wavelength of 280 nm, and peak area percentages were calculated using Empower 3 Chromatography Data System software (Waters Corporation, Milford, MA).

[0069] Particle identification (FTIR) Particle identification was performed by FTIR microscopy using a Nicolet iN10 FT-IR microscope (Thermo Fisher Scientific Inc., Massachusetts, USA). First, the sample was filtered through a gold-coated polycarbonate filter (Unchained Labs, Pleasanton, USA) with a pore size of 0.8 μm and a filtration area diameter of 4 mm. Filtration preparation involved opening the pores with several drops of filtered ethanol (0.22 μm), followed by filtering with approximately 1 mL of particle-free water as a washing step. The complete contents of each vial (previously cooled in a cold bath) were poured directly onto the filter surface. As a final step, each filter was washed with cooled particle-free water. FTIR analysis of regions of particles and undefined particles on the filter surface was performed by applying the microscopic reflection mode. Particle properties were defined by spectral comparison with internal and commercially available libraries.

[0070] Flow imaging (FlowCam) Particle morphology was characterized by flow imaging techniques using a FlowCam 8000 instrument (Fluid Imaging Technologies Inc., Scarborough, USA) with a 300 μm flow cell and 4x magnification. The system was pre-rinsed with the sample solution before each measurement. Samples were analyzed at a sampling efficiency of 75% and a flow rate of 2 mL / min.

[0071] Example 1: Pre-screening and evaluation of thermal structural protein stability in the presence of surfactants. Maximizing structural stability has been reported to improve long-term formulation quality and / or stability by preventing unfolding and aggregation of therapeutic proteins [6]. High-throughput and low-volume screening techniques involve measuring intrinsic protein fluorescence by DSC (differential scanning calorimetry) or nanoDSF (differential scanning fluorescence quantification) under isothermal chemical denaturation (ICD) or thermal denaturation conditions [7]. Thermal DSF measurements were performed to eliminate the adverse effects of surfactants on protein structural stability.

[0072] In the presence of surfactants at concentrations ranging from 0.01 mg / mL to 10 mg / mL, the stability parameters of five different mAb values ​​(bsMab1, Mab1, gMab, bsMab2, fpMab) are shown, along with the unfolding onset temperature (T). on ) and the first melting transition (T m1 The following measurements were taken. Values ​​for mAbs without surfactant addition and possessing PS20 and Px188 were obtained as a reference. In general, most of the conditions tested did not show a significant effect on the stereostructural stability of the mAbs. The data for formulations with considerable changes in their transition and melting temperatures are presented as a heatmap (Figure 1): darker colors indicate a stronger decrease in temperature due to the presence of the surfactant.

[0073] All surfactants from the chemical groups poloxamer, butron, isosorbide alkoxylate, and PVA / PPG showed no effect on the tested mAbs within the tested concentration range. However, both surfactants from the polyoxazoline group tested showed no effect on the tested mAbs. on and T m1 Compared to the reference formulations in both cases, the unstabilizing effect of fpMab and bsMab1 was observed at a concentration of 10 mg / mL.

[0074] Example 2: Pre-screening - Shaking Test The ability to protect mAbs from mechanical / interfacial stress was tested by horizontal shaking tests at a wide range of surfactant concentrations (0.001–1 mg / mL) and measured by visual inspection of visible particles (VP). Based on the data obtained by visual inspection, formulations were classified into four classes (I–IV) and shown by a heat map. A higher number of VPs resulted in a higher class and is shown in darker shades of gray. Formulations containing either PS20 or Px188, without surfactant, were used as guide reference materials to evaluate the performance of novel surfactants.

[0075] None of the tested surfactants were able to protect mAbs (fpMab) from shaking stress in the concentration range of 0.001–0.01 mg / mL. For readability, only formulations containing surfactants at 0.1 mg / mL and 1 mg / mL are shown (Figure 2).

[0076] Most surfactants showed good results at a concentration of 1 mg / mL. Here, only PVA / PPG showed a significant increase in VP numbers, while Px335, Bux016, and Pz120 showed only slight increases. Px334, Pz110, Pz120, and PVA / PPG (Class IV) at 0.1 mg / mL, as well as Bux017 (Class III) at 0.1 mg / mL, were insufficient to stabilize the formulation. Bux164, Bux190, Bux199, and IA84 also showed a slight increase in VP numbers compared to PS20. Px188 failed to protect the formulation from shaking stress at all concentrations.

[0077] Example 3: Evaluation of protein stability after long-term storage stress and heat stress in the presence of surfactants Based on data obtained from pre-screening studies, follow-up studies were conducted on the five most promising novel surfactant candidates: Px335, Bux164, Bux190, Bux199, and IA90. Under long-term storage conditions, potential adverse effects on protein stability should be excluded. Therefore, the stability of formulations with four mAbs (gMab, bsMab1, Mab1, and Mab2) was evaluated in terms of visible and microscopically visible particle formation and HMW formation under storage conditions of 9 months at 5°C, 6 months at 25°C / 60% rH, and 3 months at 40°C / 75% rH (as described in the methods) (Figures 3-6). The tested surfactant levels were kept constant at 0.06, 0.2, and 0.6 mg / mL, and PS20 or Px188 was used as a guide reference substance.

[0078] In summary, all novel surfactants performed better than or at least as well as the reference substances tested. However, there were slight differences in protection against visible particle formation (Figure 3). IA90, Bux190, and Bux199 showed good performance, comparable to or better than PS20 and Px188, but Px335 and Bux164 did not. An increase in visible particles was observed in some PS20-containing formulations, particularly in the gMab and Mab1 samples, which was likely caused by accelerated PS20 degradation and significant release of free fatty acids. The number of microscopic visible particles, measured by light shielding (Figure 6), was generally low, and the reported values ​​were USP <787> and considerably lower than the maximum acceptable number according to Ph.Eur.2.9.19. gMab and Mab1 formulations containing PS20 showed an increased amount of particles likely to be identified as free fatty acid particles based on FlowCam imaging (data not shown). Background membrane imaging results detected by Horizon (Figure 6) showed slightly higher SVP counts for Px335 and Bux164, but lower counts for Bux190, Bux199 and IA90 compared to Px188.

[0079] HMW species by SE-HPLC (Figure 5) also showed no significant changes compared to initial data. A slight increase in HMW content was detected for all boutronics after 6 months at 25°C in gMab formulations containing all novel surfactants and PS20, and after 3 months at 40°C in Mab2 formulations containing Px335 and all boutronics. This increase in HMW species was dependent on the surfactant content: the higher the concentration, the stronger the increase.

[0080] Regarding turbidity (Figure 4), it should be noted that most formulations did not show significant changes during these tests. Only the highest concentration of the novel surfactant showed a slight increase in turbidity after 3 months at 40°C, and Px335 showed a strong increase after 3 months at 40°C compared to PS20 and Px188.

[0081] One of the most significant challenges in the current use of Px188 is the formation of protein-PDMS particles (PPPs). To eliminate this, FTIR measurements were performed with two different mAbs known to readily induce PPP formation when formulated with Px188 (gMab and Mab2) on selected samples (Figure 7). No PPPs were detected with Bux190 and Bux199 formulations ≥ 0.2 mg / mL and IA90 formulation ≥ 0.6 mg / mL. Both Px188 at 0.06 and 0.2 mg / mL and Px335 at 0.06 mg / mL showed PPPs in our FTIR tests. For both Px188 and Px335, PPPs were also likely detected by FlowCam (Figure 8).

[0082] When all stability test data is included, the Bux190 and IA90 performed best, followed by the Bux199.

[0083] Example 4: Evaluation of protein stability after mechanical stress in the presence of a surfactant The effects of mechanical / interfacial stress on mAb stability were tested by stirring and freeze-thaw tests involving three different active pharmaceutical ingredients (bsMab2, IgG4, and MP). The tested surfactant levels were kept constant at 0.06, 0.2, and 0.6 mg / mL, and PS20 or Px188 was used as a guide reference substance. Stability data were obtained in terms of visible and microscopic visible particle formation, turbidity changes, and HMW species (Figures 9-12).

[0084] All surfactants, including the reference substance, showed insufficient protection of mAbs against visible particle formation, particularly at low concentrations (0.06 mg / mL) at 25°C (Figure 9). At higher concentrations (≥0.2 mg / mL), Bux164, Bux199, and IA90 performed better than or comparable to PS20. Bux190 and Px335 performed worse than PS20, and slightly better than or comparable to Px188.

[0085] The number of microscopically visible particles measured by light shielding (Figure 11) was generally at low levels, and the reported values ​​were in line with the USP Pharmacopoeia. <787> and significantly lower than the maximum acceptable number according to Ph.Eur.2.9.19. Exceptions were the bsMab2 formulation containing Px335 at all concentrations, the IgG4 formulation containing Bux190 / Px188 ≤ 0.2 mg / mL, Bux164 / Bux199 / IA90 / PS20 ≤ 0.06 mg / mL, and Bux190 at 0.06 mg / mL. Here, the particle count exceeded the detection limit. For the IgG4 formulation, no strong increase in SVP was detected, but an increase in turbidity was detected in all formulations containing 0.06 mg / mL surfactant and ≤ 0.2 mg / mL Bux190. Both the increase in SVP and IgG4 in bsMab2 indicate insufficient stabilization ability of the surfactant. Background membrane imaging results detected by Horizon (Figure 11) showed all of the higher SVP numbers, particularly at low surfactant concentrations and for Px335. However, compared to PS20, Bux164 / Bux190 and the IgG4 formulations showed higher SVP counts. Monomer content by SE-UHPLC (Figure 12) showed only slight changes in bsMab2 and MP formulations. For IgG4, a strong increase in HMW species was detected for all surfactants at ≤0.06 mg / mL and for Bux190 at ≤0.2 mg / mL, while slight increases were detected for Bux164 and Bux199 at ≤0.2 mg / mL. Bux199 and IA90 showed very good performance and were very comparable to PS20.

[0086] In summary, the novel surfactants tested here show good potential for stabilizing biological formulations compared to those currently in use. The boutronic and isosorbide alkoxylate-derived surfactants show comparable or better results than PS20 and Px188, particularly in terms of preventing protein PDMS particles. However, we have demonstrated that even within this class of compounds, there are significant differences in their ability to act as surfactants, particularly as surfactants for stabilizing aqueous antibody compositions.

[0087] Abbreviation BMI: Background Membrane Imaging Bux: Butronik DSF: Differential Scanning Fluorescence F / T: Freeze-thaw cycle FFA: Free fatty acids FTIR: Fourier Transform Infrared HMW: high molecular weight species IA: Isosorbide alkoxylate LMW: low molecular weight species mAb: Monoclonal antibody mo: Moon NTU: Turbidity Unit (Turbidity Unit by Turbidity Meter) PDMS: Polydimethylsiloxane pfs: Prefillable syringe Ph.Eur.: European Pharmacopoeia PPP: Protein-PDMS particles PS: Polysorbate Px: Poloxamer PZ: Polyoxazoline rH: Relative humidity RT: room temperature SD: standard deviation SE-(U)HPLC: Size-exclusion (ultra)fast chromatography sk:shaking Surf: Surfactant SVP: Microscope-Visible Particles Ton: Starting temperature Tm: Melting temperature USP: United States Pharmacopeia VP: Visible Particle w: Week w / o: Without

[0088] References [1] European Pharmacopeia 11.0, 2.9.20. Particulate contamination: visible particles. The European Directorate for the Quality of Medicines & Health Care, Strasbourg, France, 2022. [2] S. Kiese, A. Papppenberger, W. Friess and H.-C. Mahler, “Shaken, Not Stirred: Mechanical Stress Testing of an IgG1 Antibody,” J. Pharm. Sci., no. 97, pp. 4347 - 4366, 2008. [3] European Pharmacopeia 11.0, 2.2.1. Clarity and degree of opalescence of liquids. The European Directorate for the Quality of Medicines & Health Care, Strasbourg, France, 2022. [4] European Pharmacopeia 11.0, 2.9.19. Particulate contamination: sub-visible particles. The European Directorate for the Quality of Medicines & Health Care, Strasbourg, France, 2022. [5] “USP<787>, Subvisible particulate matter in therapeutic protein injections, Pharmacopeia Forum, 38. Pharmacopeia Forum,” 2012. [6]W.Wang,S.Nema and D.Teagarden,“Protein aggregation-pathways and influencing factors,” Int.J.Pharm.,no.390,p.89.99,2010. [7] H.Svilenov,U.Markoja and G.Winter,“Isothermal chemical denaturation as a complementary tool to overcome limitations of thermal differential scanning fluorimetry in predicting physical stability of protein formulations,”Europ.J.Pharm.and Biopharm.,no.125,p.106.113,2018.

Claims

1. An aqueous pharmaceutical composition comprising an antibody and a surfactant, wherein the surfactant is of formula (I) H-(-OE) ｎ -(OB#) ｍ -^-(BOO) ｍ -(EO) ｎ -H (I) (In the formula, -OE is ethoxy, -OBu is n-butoxy, -X- is -O-n-butylene-O- or And, n is 40, 48, or 68, and m is 11, 12, or 16. An aqueous pharmaceutical composition which is a compound of [the compound].

2. -X- is -O-n-butylene-O-, n is 40 or 48, and m is 12 or 16. The composition according to claim 1.

3. -X- is And, n is 68, and m is 11. The composition according to claim 1.

4. The composition according to any one of claims 1 to 3, wherein the antibody is a monoclonal antibody.

5. The composition according to claim 4, wherein the monoclonal antibody is of the IgG1 or IgG4 subclass.

6. The composition according to any one of claims 1 to 5, wherein the surfactant is present at a concentration of 0.001 to 1.0 mg / ml, or 0.01 to 1.0 mg / ml, or 0.06 to 1.0 mg / ml, or 0.06 to 0.6 mg / ml.

7. The composition according to any one of claims 1 to 6, further comprising additional pharmaceutically acceptable excipients.

8. Formula (I) for use as a surfactant in aqueous antibody compositions H-(-OE) ｎ -(OB#) ｍ -^-(BOO) ｍ -(EO) ｎ -H (I) (In the formula, -OE is ethoxy, -OBu is n-butoxy, -X- is -O-n-butylene-O- or And, n is 40, 48, or 68, and m is 11, 12, or 16. A compound of [this].

9. The compound of formula (I) for use according to claim 8, wherein the compound stabilizes the antibody against aggregation.

10. The compound of formula (I) for use according to claim 9, wherein the aggregation is either the aggregation of several antibodies or the aggregation of an antibody and PDMS.

11. The compound of formula (I) for use according to claim 8, wherein the compound prevents the formation of visible particles in an aqueous antibody composition.

12. -X- is -O-n-butylene-O-, n is 40 or 48, and m is 12 or 16. A compound of formula (I) for use according to any one of claims 8 to 11.

13. -X- is And, n is 68, and m is 11. A compound of formula (I) for use according to any one of claims 8 to 11.

14. A compound of formula (I) for use according to any one of claims 8 to 13, wherein the antibody is a monoclonal antibody.

15. The compound of formula (I) for use according to claim 14, wherein the monoclonal antibody is of the IgG1 or IgG4 subclass.

16. The compound of formula (I) for use according to any one of claims 8 or 15, wherein the surfactant is present at a concentration of 0.001 to 1.0 mg / ml, or 0.01 to 1.0 mg / ml, or 0.06 to 1.0 mg / ml, or 0.06 to 0.6 mg / ml.

17. A compound of formula (I) for use according to any one of claims 8 to 16, further comprising additional pharmaceutically acceptable excipients.

18. A method for preventing the formation of visible particles in an aqueous antibody composition, wherein formula (I) H-(-OE) ｎ -(OB#) ｍ -^-(BOO) ｍ -(EO) ｎ -H (I) (In the formula, -OE is ethoxy, -OBu is n-butoxy, -X- is -O-n-butylene-O- or And, n is 40, 48, or 68, and m is 11, 12, or 16. A method comprising the use of the compound.

19. -X- is -O-n-butylene-O-, n is 40 or 48, and m is 12 or 16. The method according to claim 18.

20. -X- is And, n is 68, and m is 11. The method according to claim 18.

21. The method according to any one of claims 18 to 20, wherein the antibody is a monoclonal antibody.

22. The method according to claim 21, wherein the monoclonal antibody is of the IgG1 or IgG4 subclass.

23. The method according to any one of claims 18 to 22, wherein the compound of formula (I) is present at a concentration of 0.001 to 1.0 mg / ml, or 0.01 to 1.0 mg / ml, or 0.06 to 1.0 mg / ml, or 0.06 to 0.6 mg / ml.

24. The method according to any one of claims 18 to 23, further comprising additional pharmaceutically acceptable excipients.

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